MicroRNAs expressed during normal wound healing and their associated pathways: A systematic review and bioinformatics analysis

Morgana Lüdtke Azevedo; Roberta Giorgi Silveira; Fernanda Nedel; Rafael Guerra Lund

doi:10.1371/journal.pone.0281913

. 2023 Apr 13;18(4):e0281913. doi: 10.1371/journal.pone.0281913

MicroRNAs expressed during normal wound healing and their associated pathways: A systematic review and bioinformatics analysis

Morgana Lüdtke Azevedo ^1,^*,^#, Roberta Giorgi Silveira ^2,^#, Fernanda Nedel ^2,^‡, Rafael Guerra Lund ^1,^‡,^*

Editor: Andrea Caporali³

PMCID: PMC10101427 PMID: 37053170

Abstract

MicroRNAs (miRNAs) are responsible for regulating gene expression post-transcriptionally. Are involved in several biological processes, such as wound healing. Understanding the miRNAs involved in this process is fundamental for the development of new therapies. So, due to the need to understand the role of these molecules, we aimed systematically review the literature in order to identify which miRNAs are involved in the wound healing and determine, through bioinformatics analysis, which signaling pathways are associated with these miRNAs. An electronic search was performed in the following databases: National Library of Medicine National Institutes of Health (PubMed), Science Direct, Scifinder, Scopus and Web of Science, using the descriptors: “(microRNA [MeSH])” and “(skin [MeSH])” and “(wound healing [MeSH])”. After the search, two independent and previously calibrated reviewers selected the articles that analyzed the expression pattern of miRNAs in wound healing in in vivo studies, using the software Zotero bibliography manager. Following, bioinformatic analysis was performed using the software DIANA Tools, mirPath v.3 and the data was interpreted. The bioinformatics analysis revealed that on the day 1 there were 13 union pathways, eight of which were statistically significant. Still on the day 1, among the miRNAs that had a decrease in their expression, 12 of 17 union pathways found were statistically significant. On the day 5, among the miRNAs with an increase in expression, 16 union pathways were found, 12 of which were statistically significant. Finally, among the miRNAs with decreased expression, 11 of 15 union pathways found were statistically significant. Although it has been found substantial heterogeneity in the studies, with this systematic review, it was possible to study the panorama of miRNAs that may be altered in the wound healing. The present review summarizes existing evidence of miRNAs associated to wound healing, and these findings can contribute to new therapeutic approaches.

1. Introduction

Wound healing is a biological process extremely complex and able to recover the barrier function of the skin. It requires the synchronization of several cell types beyond the interaction between cytokines and growth factors. It is composed of four different phases: hemostasis, inflammation, proliferation, and remodeling. Each one of these phases is important so that the process can occur properly. However, during the proliferative phase occurs a cascade of events of major importance for the process as a whole, such as re-epithelization, angiogenesis, and wound closure [1].

The events associated to wound healing can be affected by a variety of agents, changing the wound bed environment. Excessive wound healing–hypertrophic scar and keloid–and chronic wound are some of the sequels of impaired wound healing [2, 3]. Indeed, this process is highly complex and consists of several steps, being more susceptible to a fault. Since wound healing does not occur normally, a chronic wound may be the most recurrent consequence [4]. Nowadays, chronic non-healing wounds can affect millions of patients each year, resulting in higher morbidity and mortality of these patients [5]. Furthermore, the issue related to chronic wounds is not limited only to ulcers but is also associated with other challenges to be overcome, such as infectious and ischemic wounds. It is also known the treatment of chronic non-healing wounds remains a stretch goal on the subject of complexity and prevalence [6]. Considering this, it is clear the need for a therapeutic approach able of overcoming the problem of the complexity of this wound type in terms of treatment.

At the same time, several studies have reported miRNA as a potential therapeutic target for various diseases or conditions, from wound healing to cancer [7–9]. MiRNAs are small non-coding RNA molecules of approximately 22 nucleotides and are responsible for the regulation of gene expression at the post-transcriptional level [10, 11]. They are involved in an assorted of biological processes, from embryonic development to main cell functions, such as proliferation, differentiation, and apoptosis. The miRNA biogenesis and mechanism of action are known only about two decades ago and its molecular mechanism and is not yet totally elucidated [12, 13]. The miRNA research is clearly expanding, mainly in the last five years, because increasingly more studies have elucidated some points of miRNA molecular mechanisms. Moreover, as the miRNAs are important regulators of gene expression, may be promising targets for the development of biomarkers and can help in the development of a therapy [14].

Although there are still some questions about miRNA to be clarified, it is currently known that it is a key part of the entire epigenetic machinery acting as an important regulator of gene expression [15]. The epigenetic plays an important role in all the processes that occur in living organisms and can be used to explain several features of diseases and biological events, such as their pathway or late onset and end [16]. However, miRNAs are not only a part of the epigenetic machinery but are also can modify epigenetically by DNA methylation and histone modification just like another protein-coding gene [17, 18]. These changes in the miRNA expression pattern occur also during the wound healing, and as reported several miRNAs can be found decreased, including the members of the miR-200 family [19], or increased like miR-31, miR-33 and miR-196, among others [20–23]. Also was reported that certain miRNAs, such as miR-21, can mitigate possible aging-associated wound healing failures [24].

Therefore, comprehension of the miRNAs role in wound healing, as well as where and how it acts, can be applied in identifying the pathways involved in the process in order to bring new possibilities of molecular target therapies to the defects that can occur during wound healing. Further, considering the lack of studies about the panorama of miRNAs and their signaling pathways, mainly related to wound healing, we aimed to systematically review the available literature for the purpose of identifying which miRNAs are associated with the wound healing phases and their associated pathways using bioinformatics analysis.

2. Material and methods

2.1 Registration and review questions

The current systematic review is reported in accordance with the Preferred Items for Systematic Review and Meta-Analysis (PRISMA) guidelines. Due to the study design’s nature, the protocol was registered in the Open Science Framework and is available at the following link (https://osf.io/xp4tf/).

The review questions were: (1) Which miRNAs have been related to the wound healing process? (2) In which pathways do these miRNAs act?

2.2 Inclusion and exclusion criteria

The inclusion criterion for the articles was in vivo animal studies that analyzed the miRNA expression patterns in the wound healing process. The choice to use only in vivo animal studies is due to the similarity found in the methodology of these studies, reducing possible confounding variables. Only studies published in English were included. The exclusion criteria were congress summary, book section, literature reviews, hypothesis articles, opinion articles, methodological approaches, commentaries, previews, expert opinions, letters, news, patents, studies unrelated to miRNA and wound healing, and studies with confounding factors. Besides that, articles that were not written in English and not fully available were also excluded.

2.3 Search strategy

The electronic strategy was carried out without initial date restriction up to and including May 2020 in the following databases: PubMed, Science Direct, Scifinder, Scopus, and Web of Science. In this case, the Google Scholar database was not used due to the fact that it presents low precision in recently established themes, such as miRNAs [25, 26]. The search strategy was conducted using the following terms: “(MicroRNAs) AND (Skin) AND (Wound Healing)”. No language was applied in the search.

2.4 Study selection

The search results have been exported to the Zotero bibliography manager. Initially, duplicate records were excluded. Titles, abstracts, and study methodologies were screened based on the inclusion and exclusion criteria by two blinded and independent reviewers (MLA and RGS). All records were compared and in case of disagreement, a consensus was reached by discussion. When consensus was not achieved, a different reviewer decided if the article should be included (RGL).

2.5 Data extraction

Data were extracted and tabulated independently by two reviewers (MLA and RGS) in an Excel spreadsheet (Microsoft Corporation, Redmond, WA, USA) to be submitted for descriptive analysis. Cases of disagreement were handled as described above.

2.6 Bioinformatics analysis

After extracting the data, the miRNAs that showed different expression patterns (higher or lower expression) and were repeated on days 1 and 5 during the wound healing process were separately inserted, using the software DIANA Toll mirPath v.3. The inclusion criterion for bioinformatics analysis were the miRNAs that were described more than one time at different times analyzed in the selected studies, and the exclusion criterion was the miRNAs described just one time in the selected studies beyond the miRNAs do not index in mirPath v.3. So, the miRNAs that were not recognized by the software were disregarded in this analyze.

The analyses were performed in real-time using the Kyoto Encyclopedia of Genes and Genoma (KEGG) selecting the murine specie to investigate the miRNAs. The interactions dataset chosen for all miRNAs was TarBase v7.0 and in the advanced statistics options, False Discovery Rate (FDR) correction was chosen. The Fisher’s Exact Test was used with a p-value threshold of 0.05. After inserting all miRNAs into the software, the pathway associated were observed and tabulated, specifying its p-value and its relationship to the wound healing process.

2.7 Quality assessment

To assess the quality of the studies included in this systematic review, the Review Manager 5.4.1 software was used. The checklist was composed of three domains: wound healing assay, miRNA analysis, and the presence of a control group (uninjured skin). Two independent researchers (MLA and RGS) assessed the quality of the studies based on criteria previously established. In cases of disagreement were discussed until a consensus was reached, and when a consensus was not obtained, a third reviewer participated in the discussion (RGL).

3. Results

The initial search amounted to 3,227 records. After the removal of duplicates, a total of 2,370 articles remained for the title and abstract screening. So, based on the title and abstract, 915 articles remained for full-text reading screening. After reading the full text, were excluded 847 articles, remaining 68 articles satisfy the inclusion criteria. These of 68 articles, 40 were excluded for not performing the analysis in vivo animal. A total of 28 articles remained, and 16 of these ended up being excluded for showing possible confounding variables, such as studies that used some type of treatment or transgenic animal. In other words, studies did not analyze wound healing under regular conditions. Finally, remained 12 articles that present viable data to analyze, as shown in the PRISMA flowchart for the study selection process (Fig 1).

Fig 1 — Flow diagram of the study selection process according to PRISMA Statement. * A Study could have fulfilled more than one criterion.

Among the 12 selected studies, it is clear that the country of publication of the most recurrent articles is the USA, totaling 6 of the 12 articles, followed by China with 4 of these publications. Regarding the year of publication, the oldest article date from 2012, demonstrating how recent the miRNAs studies, especially in relation to wound healing, is recent. These results are also described and illustrated in the figure below (Fig 2).

Fig 2 — Correlation between publication countries and years of selected articles.

Table 1 refers to the main methodologies used in the 12 selected articles. Concerning the animal model used, the choice was unanimous. Absolutely all articles used mice as an animal model, there were a few studies that used rats; however, these ended up being excluded due to eligibility criteria. The most frequent strains were C57BL/6, C57BL/6J, and Balb-C. Interestingly, the SKH-1 strain, which is hairless mice in the skin, was reported in only one article. Almost half of the studies specified the gender of their animals–female (n = 3) and male (n = 4)–the other half did not specify. Most animals were between 8–10 weeks old. The more frequent methodology applied for the induction of the lesion was full-thickness wound by biopsy punch in 100% of the articles. The recurrent induction site of the lesion was on the dorsal skin, and the lesions were sized between 3–6 mm.

Table 1. Correlation of the main methodological approaches found in the selected articles.

Author, Year	Animal conditions				Wound procedures
	Animal	Strain	Sex	Age	Injury induction site	Injury type	Injury size
Aunin et al., 2017	Mice	-	-	8 weeks	Back skin	Full-thickness wound by biopsy punch	3 mm
Aunin et al., 2017	Mice	-	-	2 years	Back skin	Full-thickness wound by biopsy punch	3 mm
Chan et al., 2012a	Mice	C57BL/6	Male	8–10 weeks	Dorsal skin	2 or 4 full-thickness excisional wounds by biopsy punch	6 mm
Chan et al., 2012b	Mice	C57BL/6	Male	-	Dorsal skin	2 full-thickness wound	8x16 mm
Chan et al., 2012b	Mice	C57BL/6	Male	-	Dorsal skin	2 or 4 full-thickness excisional wounds by biopsy punch	6 mm
Chen et al., 2019	Mice	Balb/c	Female	8–10 weeks	Dorsal skin	2 full-thickness incisional wounds using a pair of scissors	10 mm
					Dorsal skin	2 full-thickness excisional wounds by biopsy punch	5 mm
					Anterior of the hard palate	3 incisional wounds using a scalpel blade	50 mm
Etich et al., 2017	Mice	C57BL/6N	-	8–10 weeks	Back skin	Full-thickness wounds by biopsy punch	6 mm
Jin and Chung, 2018	Mice	SKH-1	Female	8 weeks	Dorsal skin	2 full-thickness wounds by biopsy punch	3.5 mm
Shi et al., 2018	Mice	-	-	7–8 weeks	Dorsal skin	Full-thickness wounds by biopsy punch	6 mm
Shi et al., 2018	Mice	K14-Cre	-	7–8 weeks	Dorsal skin	Full-thickness wounds by biopsy punch	6 mm
Simões et al., 2019	Mice	Balb/c	Female	8–10 weeks	Dorsal skin	Full-thickness wounds by biopsy punch	5mm
					Hard palate from maxilla	Full-thickness with a pair of forceps	-
					Dorsal skin	2 full-thickness excisional wounds by biopsy punch	5 mm
van Solingen et al., 2014	Mice	B6.Cg-Mirn155^tm1.1Rsky/J	-	10–12 weeks	-	2 full-thickness by biopsy punch	6 mm
van Solingen et al., 2014	Mice	C57BL	-	10–12 weeks	-	2 full-thickness by biopsy punch	6 mm
Wang et al., 2012	Mice	C57BL	Male	Adult	Back	Full-thickness skin excision	10 mm
Wang et al., 2019	Mice	C57BL (H-2b)	Male	Adult	Back	2 full-thickness by biopsy punch	6 mm
Wang et al., 2019		B6.Cg-Mirn155^tm1.1Rsky/J	Male	6–8 weeks	Back	2 full-thickness by biopsy punch	6 mm
Zhao et al., 2020	Mice	C57BL/6J	-	8–12 weeks	Back	2 circular full-thickness excisional wounds	6 mm
		C57BL/6J	-	8–12 weeks	Back	2 circular full-thickness excisional wounds	6 mm
		-	Male and female	-	Back	2 circular full-thickness excisional wounds	6 mm

Open in a new tab

Main findings regarding the methodological approaches of the studies include animal and strain, injury induction site, injury size, and wound procedure.

As shown in S1 Table, the main techniques used to analyze the expression of miRNA were RT-qPCR, reported by most articles, and microarray. Some studies combined the two methodologies. The total amount of miRNA that had its expression analyzed varied according to the days of analysis. Among the most recurrent analysis times that were related to the wound healing phases, three days stood out: day 1, day 3, and day 5. However, the relationship between the down and up expression on day 3 was verified by only one of the selected articles (Aunin et al., 2017). The other articles found alteration of the expression in only one of these two patterns and not concurrently (Chan et al., 2012 (a), Chan et al., 2012 (b) and Zao et al., 2020 found alterations in just decreased expression. While, Shi et al., 2018 and Wang et al, 2019 found alterations in just increased expression). So the bioinformatics analysis was performed according to the data obtained on days 1 and 5.

On day 1, a total of 273 miRNAs were analyzed, and just two of these were not affected, in other words, do not have alteration in their expression pattern. Among these that showed some alteration in expression pattern, 153 miRNAs had an increase in their expression, while 118 miRNAs, decreased. On day 5, the time of analysis with the highest amount of analyzed miRNA, the not affected ones totaled 5 miRNAs, while those with increased and decreased expression were 192 and 130, respectively. At last, considering these two days of analysis of the 12 selected articles, a total of 600 miRNAs had their expression pattern analyzed during the wound healing process. The most frequently found miRNAs on days 1 and 5 that were analyzed by the bioinformatic are described in Table 3.

Table 3. Statistically significant union pathways refer to the miRNAs with a decreased expression on day 1.

Pathway	p-value	Target genes
Prion diseases	6.72018e-13	7
Hippo signaling pathway	4.282252e-09	1
Proteoglycans in cancer	2.17414e-07	59
Caffeine metabolism	1.547323e-05	2
Renal cell carcinoma	9.472664e-05	17
Steroid biosynthesis	0.0001140205	5
Lysine degradation	0.00122778	18
Adherens junctions	0.001282294	20
TGF-beta signaling pathway	0.00177966	13
FoxO signaling pathway	0.003806427	47
Endocytosis	0.004723869	55
N-Glycan biosynthesis	0.0115209	15
Drug metabolism–other enzymes	0.01724572	6
Protein processing in endoplasmatic reticulum	0.0386971	55
Cicardian entrainment	0.04087073	1
Morphine addiction	0.04087073	1
Pancreatic cancer	0.04101645	23

Open in a new tab

The bioinformatics analysis discloses that on the day 1, among the miRNAs that had their expression increased, there were three intersection pathways, but none with a statistically significant difference (p ≤ 0.05) and 13 union pathways, eight of which were statistically significant. In the miRNAs that had a decrease in their expression, 17 union pathways were found, 12 of which were statistically significant, and no one intersection pathway was found. On the day 5, in the miRNAs with an increase in expression, 16 union pathways were found, 12 of which were statistically significant, and also no one intersection pathway was found. Among the miRNAs with decreased expression, 15 union pathways were found, 11 of which were statistically significant, and no one intersection pathway was found (Tables 2–5), and signaling pathways related to the wound healing process are highlighted in bold.

Table 2. Statistically significant union pathways refer to the miRNAs with an increased expression on day 1.

Pathway	p-value	Target genes
Fatty acid biosynthesis	<1e-325	2
Fatty acid metabolism	6.165574e-09	4
Steroid biosynthesis	6.921681e-08	3
Central carbon metabolism in cancer	2.532531e-05	9
Caffeine metabolism	2.639553e-05	2
Proteoglycans in cancer	0.009315449	8
Carbohydrate digestion and absorption	0.01156247	4
Fat digestion and absorption	0.01410009	2
Complement and coagulation cascades	0.04045149	7
Thyroid hormone and signaling pathway	0.04320222	11
HIF-1 signaling pathway	0.04440003	7
Renal cell carcinoma	0.04873515	7
Viral carcinogenesis	0.0496533	11

Open in a new tab

Table 5. Statistically significant union pathways refer to the miRNAs with a decreased expression on day 5.

Pathway	p-value	Target genes
Hippo signaling pathway	2.384482e-11	1
Caffeine metabolism	8.713539e-08	2
Proteoglycans in cancer	7.888269e-06	32
Drug metabolism–other enzymes	0.0001989493	6
Steroid biosynthesis	0.0003114483	2
Cicardian entrainment	0.0006915614	1
Morphine addiction	0.0006915614	1
Lysine degradation	0.001772916	12
Metabolism of xenobiotics by cytochrome P450	0.00374839	4
Endocytosis	0.005002083	34
ErbB signaling pathway	0.008312138	18
MAPK signaling pathway	0.01457244	39
Phosphatidylinositol signaling system	0.01930813	16
Colorectal cancer	0.02517332	14
Protein processing in the endoplasmic reticulum	0.04973187	29

Open in a new tab

Correlation of binding pathways observed in miRNAs with increased and decreased expression and their respective target genes.

Table 4. Statistically significant union pathways refer to the miRNAs with an increased expression on day 5.

Pathway	p-value	Target genes
Fatty acid biosynthesis	<1e-325	2
Fatty acid metabolism	4.440892e-16	11
Renal cell carcinoma	1.630683e-08	23
Steroid biosynthesis	3.491662e-07	5
TGF-beta signaling pathway	0.0001216682	13
Lysine degradation	0.0003168722	10
Thyroid hormone signaling pathway	0.0005841115	25
FoxO signaling pathway	0.0007019681	31
Central carbon metabolism in cancer	0.003639735	6
Proteoglycans in cancer	0.01120219	19
HIF-1 signaling pathway	0.01131124	17
Viral carcinogenesis	0.01320044	26
Bacterial invasion of epithelial cells	0.0176033	17
RNA degradation	0.0310473	10
Regulation of actin cytoskeleton	0.03361088	37
Axon guidance	0.04726244	25

Open in a new tab

The miRNAs and their associated pathways also are illustrated in Fig 3. In the Venn diagrams (Fig 4) it is possible to observe the signaling pathways that overlap on both days (1 and 5), as well as those that occur exclusively on either day 1 or day 5.

Based on the quality assessment of the 12 studies, the three pre-established domains proved to be adequate (Fig 5). The requirement to perform the miRNA analysis in addition to the presence of a control group were the domains that presented insufficient or absent definitions according to pre-established domains.

4. Discussion

In this systematic review, based on the results obtained, can be observed a panorama of altered miRNAs during the wound healing process and their pathways associated (Table 2).

Recent studies have often reported the importance of the role of miRNA in several cellular processes, including wound healing. These studies claim that during the process the miRNA expression pattern can alternate according to the day, suggesting its regulatory function in these cases [27–29]. In our results, it was possible to observe these patterns on different days of wound healing. on each of the days analyzed, essentially all the analyzed miRNAs suffered some alteration in their expression pattern. The miRNAs found with some alterations were quite varied, considering all analysis times.

On day 1, among the miRNAs with an increase in their expression (mmu-miR-223-3p and mmu-miR-34c-5p), already it is possible to observe the correlation with wound healing. Evidence had shown that mmu-miR-223-3p can ameliorates vascular endothelial lesions through the IL6ST and STAT3 signaling pathways [23, 30]. Furthermore, members of the miR-223 family have been associated as important regulators in the inflammatory process that occurs during early phases of wound healing, which justifies its increase in expression on day 1, when the inflammatory process of tissue repair occurs. Also, the miR-223 family is effective in increasing the activation of neutrophils after episodes of bacterial infections, and consequently, improving the course of the healing process [31, 32]. The miR-34 family also seems to be related to wound healing; it has been reported that several family members are up-regulated in epidermal keratinocytes during wound healing. In addition, it being able to improve the inflammatory process occurring during healing due to the release of inflammatory cytokines [33, 34].

The miR-31 family is also closely related to the wound healing process, these miRNAs can regulate keratinocytes proliferation, differentiation, and migration through the regulation of the signaling pathways NF-κB, RAS/MAPK, Notch, and cytokines [35, 36]. The miR-31-5p was found with expression decreased on day 1 and increased on day 5. Already described for its important role in cell migration, the miR-199a-5p was found decreased on both days [37], but also there is evidence that miR-199a-5p can, in a negative way, regulate the angiogenic responses through responses by directly targeting ETS proto-oncogene 1 and transcription factor (ETS-1) [38]. The first miRNAs associated with inflammatory response were miR-132 and miR-125b, the expression was induced in a monocytic cell line treated with lipopolysaccharide [39–41]. However, the miR-125a-5p and miR-125b-5p were found with a decreased expression on both days. The angiogenesis also is regulated by the miRNAs. For instance, some studies already related the downregulation of miR-199a-5p expression in the dermis and endothelial tissue during the wound healing process. Also, by targeting an angiogenesis-related transcription factor and its mediator, the miR-199a-5p can, negatively, regulate the angiogenic response of dermal microvascular endothelial cells in humans. In mice with homozygous deletions in the ETS-1 gene, was related to an impaired of angiogenesis, insufficient formation of granulation tissue, and compromised wound closure [41–43]. The miRNA-199a-5p was found with an expression decreased on day 1, but not on day 5.

In our bioinformatics analysis, we investigated the associated pathways highlighting the statistically significant and related to wound healing. Among them, the thyroid hormone signaling pathway (p = 0.04) was identified. This pathway can be related to several biological processes by regulating gene expression. The thyroid hormone, for instance, already was described as one of the most potent stimulators of growth and metabolic rate, it can induce the angiogenesis through the increase of bGFG mRNA expression via the integrin αvβ3/PKD/HDAC5 signaling pathway [44, 45]. In a culture of human keratinocytes, the exogenous thyroid human stimulated the expression of keratin genes, these genes are responsible for 30% of the protein of the epidermis, and there is clear evidence about the relation to keratin genes and wound healing specific phases [46]. Another representative pathway associated with miRNAs with an increased expression is the Hypoxia Inducible Factor-1 (HFI-1) signaling pathway (p = 0.04). Several studies already related the role of HIF-1 in wound healing, contributing to cell migration and division under hypoxic conditions, beyond the growth factor release and extracellular matrix [47, 48].

The forkhead box O (FoxO) signaling pathway was also found among the miRNAs with an increased expression (p = 0.0007) targeting more than 30 genes. The FoxO family is constituted of transcription factors responsible for regulating gene expression in several cellular events and biological processes, such as apoptosis, cell-cycle control, oxidative stress resistance and wound healing stimulation [49]. Also, the members of this family are recruited for keratinocyte mobilization and migration due to their ability to regulate, positively, transforming growth factor-beta (TGF-β1) expression. The TGF-β1 exerts effects on wound healing through immune modulation, cell proliferation, migration and differentiation regulation, and extracellular matrix production [50]. The TGF-β1 signaling pathway was also found associated to wound healing (p = 0.0001). There are three isoforms (TGF-β1, TGF-β2 and TGF-β3) and all of these appears to exert effects on wound healing through the SMAD pathway, mainly. The TGF-β1 is more frequently related to scarless wound healing formation, whereas the TGF-β3 already was observed in fibrotic scarring [51]. These findings corroborate the TGF-β1 decrease described in chronic non-healing wounds [52]. In general, the TGF-β family can play a role he wound healing through inflammation regulation, fibroblast proliferation, angiogenesis simulation, and deposition and remodeling of the extracellular matrix [53].

Among the signaling pathways associated with the miRNAs with decreased expression and related to the wound healing process, was found the steroid biosynthesis pathway (p = 0.0001). This signaling pathway can be related to wound healing in many ways; glucocorticoids (GC) are able to inhibit wound healing through their membranous glucocorticoid receptor. This receptor via activation of the Wnt-like 6 PLC/PKC signaling cascade interferes with the keratinocytes migration and, consequently, on wound closure [54]. The Wnt signaling pathway already was, frequently, related to several aspects of skin development and physiology. Also, in cases of Wnt pathway reduction, the regenerative capacities and abilities are impaired. This pathway regulates the β-catenin activation, and this process appears to be one of the several inflammatory responses to injury [54, 55]. Although the Wnt signaling pathway in the inflammatory response is not yet very understood, some evidence, through the observation of gene Wnt5 increased expression in patients with severe sepsis, suggests β-catenin-independent Wnt signaling may be a proinflammatory stimulus, a key event for the wound healing process [56].

Cell adhesion, mediated by adherens junctions, is crucial and closely related to the wound healing process. The adherens junction signaling pathway (p = 0. 001) plays an important role in cell plasticity, providing both cell-cell adhesion and fast cell-cell contact remodeling during several biological processes, such as wound healing [57]. The adherens junctions are also a key target of endocytosis during wound healing; the endocytosis signaling pathway (p = 0.004), also found in miRNAs with decreased expression, provides the endocytic remodeling of adherens junctions. This cell adhesion is required to control the actin assembly on the wound edge increasing the speed of wound closure [58]. There is some evidence suggesting that most cell-cell adhesion proteins can be modified by N-Glycans, increasing the probability of the occurrence of defects in the formation of the protective barrier and cell differentiation and adhesion [59]. The N-Glycans biosynthesis signaling pathway (p = 0.01), and these glycans are clearly involved in processes responsible for regulating the terminal differentiation products in keratinocytes [60].

In addition, were identified two signaling pathways among the miRNAs with decreased expression involving the protein kinase. The ErbB (epidermal growth factor receptor) signaling pathway (p = 0.08) is a family of receptor tyrosine kinases (RTKs) responsible for binding extracellular growth factor to intracellular pathways, to regulate some biological responses–cell proliferation, differentiation, and motility [61]. These growth factors, acting via RTKs, are able to control different cell types in skin wound healing, particularly macrophages and neutrophils. For this reason, the aberrant expression of the growth factors or their receptors is related to difficult wound healing [62]. Cell proliferation is also regulated for the mitogen-activated protein kinase (MAPK) signaling pathway (p = 0.01). Through more than 18 target genes, the activation of the MAPK pathway, mainly the extracellular signal-regulated kinase (ERK), is the most important regulator of several cell types’ migration. The ERK/MAPK pathway can be activated by skin damage and this activation has a strong effect on keratinocyte migration [63–66].

Ultimately, this systematic review has limitations that need to be highlighted. Different strains of mice were used to observe the miRNA pattern expression during wound healing, although there are certain strains most frequently used, their differences must be considered so that the results are properly interpreted. In different strains, beside the miRNA pattern expression, the signaling pathways also can be altered [67, 68]. Another variable that can induce confusion and doubts is the analysis time. Wound healing is a complex and extensive process, in other words, does not occur in a fixed time, but in three or four phases taking to days to months [1, 3]. Thus, it is necessary to define the most adequate analysis time to obtain the most reliable data possible, according to the studied species and the biological processes that the objective is to study. Nonetheless, in some cases, such heterogeneity may be important, for example, to identify new pathways that would not be related otherwise.

Hence, the results found in this systematic review and bioinformatics analysis contribute to the identification of miRNAs altered during the wound healing process, as well as the associated signaling pathways. Furthermore, they can be used as a study tool for the next works that aim to relate miRNAs to wound healing, in the search for new potential biomarker targets, whether for diagnosis or therapy.

5. Conclusion

In conclusion, the results of our systematic review demonstrated that some miRNAs are altered during the wound healing process. The bioinformatics analysis revealed that on day 1, among the miRNAs with increased and decreased expression, there are 20 union pathways that were statistically significant. And on day 5, we can observe a similar amount, totalizing 23 union pathways statistically significant. Most miRNAs identified in our study play a role in wound healing through regulating, mainly, cell proliferation and differentiation, by several signaling pathways. It is worth noting the limitation we found in the selected studies regarding their significant heterogeneity, which can be explained by the differences in the target populations—in this case, the wound healing is observed in different strains of mice -, and timing of outcome measurements. In this sense, even though there is a diversity in the studies found, this heterogeneity can be important in order to identify new pathways that would probably not be considered in another manner, as in studies with low heterogeneity. But even then, the results we present help to better understand the complex network of miRNAs, as well as their role in the healing of wounds. With this systematic review, it was possible to study the panorama of miRNAs that may be altered in the wound healing, understanding which miRNAs and its respective signaling pathways may be involved in the wound healing process. The present review summarizes existing evidence of miRNAs associated to wound healing, and these findings can contribute to new therapeutic approaches.

Supporting information

S1 Checklist. PRISMA 2020 checklist.

(DOCX)

Click here for additional data file.^{(31.4KB, docx)}

S1 Table. Correlation of results regarding the expression pattern of miRNAs at different times of analysis.

(DOCX)

Click here for additional data file.^{(73.6KB, docx)}

Data Availability

All relevant data are within the paper.

Funding Statement

The authors received no specific funding for this work.

References

1.Rodrigues M, Kosaric N, Bonham CA, Gurtner GC. Wound healing: a cellular perspective. Physiol Rev. 2019;99: 665–706. doi: 10.1152/physrev.00067.2017 [DOI] [PMC free article] [PubMed] [Google Scholar]
2.Basu S, Shukla V. Complications of wound healing. In: Measurements in wound healing. Springer. 2012: 109–144. [Google Scholar]
3.Wang PH, Huang BS, Horng HC, Yeh CC. Wound healing. J Chin Med Assoc. 2017;20: 1–8. [DOI] [PubMed] [Google Scholar]
4.Han G, Ceilley R. Chronic wound healing: a review of current management and treatments. Adv Ther. 2017;34: 599–610. doi: 10.1007/s12325-017-0478-y [DOI] [PMC free article] [PubMed] [Google Scholar]
5.Yao Z, Niu J, Cheng B. Prevalence of chronic skin wounds and their risk factors in an inpatient hospital setting in northern China. Adv Wound Care. 2020;33: 1–10. [DOI] [PubMed] [Google Scholar]
6.Gupta S, Andersen C, Black J, de Leon J, Fife C, John CL, et al. Management of Chronic Wounds: Diagnosis, Preparation, Treatment, and Follow-up. Wounds. 2017;29: 19–36. [PubMed] [Google Scholar]
7.Christopher AF, Kaur RP, Kaur G, Kaur A, Gupta V, Bansal P. MicroRNA therapeutics: Discovering novel targets and developing specific therapy. Perspect Clin Res. 2016;7: 68. doi: 10.4103/2229-3485.179431 [DOI] [PMC free article] [PubMed] [Google Scholar]
8.Rupaimoole R, Slack FJ. MicroRNA therapeutics: towards a new era for the management of cancer and other diseases. Nat Rev Drug Discov. 2017;16: 203–222. doi: 10.1038/nrd.2016.246 [DOI] [PubMed] [Google Scholar]
9.Soliman AM, Das S, Abd Ghafar N, Teoh SL. Role of microRNA in proliferation phase of wound healing. Front genet. 2018;9: 38. doi: 10.3389/fgene.2018.00038 [DOI] [PMC free article] [PubMed] [Google Scholar]
10.Hanna J, Hossain GS, Kocerha J. The potential for microRNA therapeutics and clinical research. Front genet. 2019;10: 478. doi: 10.3389/fgene.2019.00478 [DOI] [PMC free article] [PubMed] [Google Scholar]
11.Catalanotto C, Cogoni C, Zardo G. MicroRNA in control of gene expression: an overview of nuclear functions. Int J Mol Sci. 2016;17: 1712. doi: 10.3390/ijms17101712 [DOI] [PMC free article] [PubMed] [Google Scholar]
12.Creugny A, Fender A, Pfeffer S. Regulation of primary micro RNA processing. FEBS Lett. 2018;592: 1980–1996. doi: 10.1002/1873-3468.13067 [DOI] [PubMed] [Google Scholar]
13.Macgregor-Das AM, Das S. A microRNA’s journey to the center of the mitochondria. Am J Physiol Heart Circ Physiol. 2018;315: 206–215. [DOI] [PubMed] [Google Scholar]
14.Lu TX, Rothenberg ME. MicroRNA. J Allergy Clin Immunol. 2018;141: 1202–1207. doi: 10.1016/j.jaci.2017.08.034 [DOI] [PMC free article] [PubMed] [Google Scholar]
15.Piletič K, Kunej T. MicroRNA epigenetic signatures in human disease. Arch Toxicol. 2016;90: 2405–2419. doi: 10.1007/s00204-016-1815-7 [DOI] [PubMed] [Google Scholar]
16.Yao Q, Chen Y, Zhou X. The roles of microRNAs in epigenetic regulation. Curr Opin Chem Biol. 2019;51: 11–17. doi: 10.1016/j.cbpa.2019.01.024 [DOI] [PubMed] [Google Scholar]
17.Cui J, Zhou B, Ross SA, Zempleni J. Nutrition, microRNAs, and human health. Adv Nutr. 2017;8: 105–112. doi: 10.3945/an.116.013839 [DOI] [PMC free article] [PubMed] [Google Scholar]
18.Saliminejad K, Khorram Khorshid HR, Soleymani Fard S, Ghaffari SH. An overview of microRNAs: biology, functions, therapeutics, and analysis methods. J Cell Physiol. 2019;234: 5451–5465. doi: 10.1002/jcp.27486 [DOI] [PubMed] [Google Scholar]
19.Aunin E, Broadley D, Ahmed MI, Mardaryev AN, Botchkareva NV. Exploring a role for regulatory miRNAs in wound healing during ageing: involvement of miR-200c in wound repair. Sci Rep. 2017;7: 1–10. [DOI] [PMC free article] [PubMed] [Google Scholar]
20.Chan YC, Roy S, Huang Y, Khanna S, Sem CK. The microRNA miR-199a-5p down-regulation switches on wound angiogenesis by derepressing the v-ets erythroblastosis virus E26 oncogene homolog 1-matrix metalloproteinase-1 pathway. J Biol Chem. 2012;287: 41032–41043. doi: 10.1074/jbc.M112.413294 [DOI] [PMC free article] [PubMed] [Google Scholar]
21.Etich J, Bergmeier V, Pitzler L, Brachvogel B. Identification of a reference gene for the quantification of mRNA and miRNA expression during skin wound healing. Connect Tissue Res. 2017;58: 196–207. doi: 10.1080/03008207.2016.1210606 [DOI] [PubMed] [Google Scholar]
22.Shi J, Ma X, Su Y, Song Y, Tian Y, Yuan S, et al. MiR-31 mediates inflammatory signaling to promote re-epithelialization during skin wound healing. J Investig Dermatol. 2018; 138: 2253–2263. doi: 10.1016/j.jid.2018.03.1521 [DOI] [PMC free article] [PubMed] [Google Scholar]
23.Wang CR, Zhu HF, Zhu Y. Knockout of microRNA-155 ameliorates the Th17/Th9 immune response and promotes wound healing. Curr Med Sci. 2019;39: 954–964. doi: 10.1007/s11596-019-2128-x [DOI] [PubMed] [Google Scholar]
24.Long S, Zhao N, Ge L, Wang G, Ran X, Wang J, et al. MiR‐21 ameliorates age‐associated skin wound healing defects in mice. J Genet Med. 2018;20: 3022. doi: 10.1002/jgm.3022 [DOI] [PubMed] [Google Scholar]
25.Boeker M, Vach W, Motschall E. Google Scholar as replacement for systematic literature searches: good relative recall and precision are not enough. BMC Med Res Methodol. 2013;13: 1–12. [DOI] [PMC free article] [PubMed] [Google Scholar]
26.Silveira RG, Ferrúa CP, do Amaral CC, Garcia TF, de Souza KB, Nedel, F. MicroRNAs expressed in neuronal differentiation and their associated pathways: systematic review and bioinformatics analysis. Brain Res Bull. 2020;157: 140–148. [DOI] [PubMed] [Google Scholar]
27.Wang W, Yang C, Wang X, yan Zhou L, Juan Lao G, Liu D, et al. MicroRNA-129 and-335 promote diabetic wound healing by inhibiting Sp1-mediated MMP-9 expression. Diabetes. 2018;67: 1627–1638. doi: 10.2337/db17-1238 [DOI] [PubMed] [Google Scholar]
28.Jiang Z, Wei J, Yang W, Li W, Liu F, Yan X, et al. MicroRNA-26a inhibits wound healing through decreased keratinocytes migration by regulating ITGA5 through PI3K/AKT signaling pathway. Biosci Rep. 2020;40: BSR20201361. doi: 10.1042/BSR20201361 [DOI] [PMC free article] [PubMed] [Google Scholar]
29.Wu Y, Zhang K, Liu R, Zhang H, Chen D, Yu S, et al. MicroRNA-21-3p accelerates diabetic wound healing in mice by downregulating SPRY1. Aging. 2020;12: 15436. doi: 10.18632/aging.103610 [DOI] [PMC free article] [PubMed] [Google Scholar]
30.Zanoaga O, Braicu C, Chiroi P, Andreea N, Hajjar NA, Mărgărit S, et al. The Role of miR-155 in Nutrition: Modulating Cancer-Associated Inflammation. Nutrients. 2012;13(7): 2245. [DOI] [PMC free article] [PubMed] [Google Scholar]
31.Wolska-Gawron K, Bartosińska J, Rusek M, Kowal M, Raczkiewicz D, Krasowska D. Circulating miRNA-181b-5p, miRNA-223-3p, miRNA-210-3p, let 7i-5p, miRNA-21-5p and miRNA-29a-3p in patients with localized scleroderma as potential biomarkers. Sci Rep. 2020;10: 1–14. [DOI] [PMC free article] [PubMed] [Google Scholar]
32.Córdova-Rivas S, Fraire-Soto I, Mercado-Casas Torres A, Servín-González LS, Granados-López AJ, López-Hernández Y, et al. 5p and 3p strands of miR-34 family members have differential effects in cell proliferation, migration, and invasion in cervical cancer cells. Int J Mol Sci. 2019;20: 545. doi: 10.3390/ijms20030545 [DOI] [PMC free article] [PubMed] [Google Scholar]
33.Wu J, Li X, Li D, Ren X, Li Y, Herter EK, et al. MicroRNA-34 family enhances wound inflammation by targeting LGR4. J Investig Dermatol. 2020;140: 465–476. doi: 10.1016/j.jid.2019.07.694 [DOI] [PubMed] [Google Scholar]
34.Childs DR, Murthy AS. Overview of wound healing and management. Surgical Clinics. 2017;97: 189–207. doi: 10.1016/j.suc.2016.08.013 [DOI] [PubMed] [Google Scholar]
35.Chen L, Simões A, Chen Z, Zhao Y, Wu X, Dai Y, et al. Overexpression of the oral mucosa-specific microRNA-31 promotes skin wound closure. Int J Mol Sci. 2019;20: 3679. doi: 10.3390/ijms20153679 [DOI] [PMC free article] [PubMed] [Google Scholar]
36.Wang F, Gao Y, Yuan Y, Du R, Li P, Liu F, et al. MicroRNA-31 Can Positively Regulate the Proliferation, Differentiation and Migration of Keratinocytes. Biomed Hub. 2020;5(2): 1–12. doi: 10.1159/000508612 [DOI] [PMC free article] [PubMed] [Google Scholar]
37.Yan S, Xu Z, Lou F, Zhang L, Ke F, et al. NF-κB-induced microRNA-31 promotes epidermal hyperplasia by repressing protein phosphatase 6 in psoriasis. Nat Commun. 2015;6(1): 1–15. [DOI] [PMC free article] [PubMed] [Google Scholar]
38.Gheinani AH, Burkhard FC, Rehrauer H, Fournier CA, Monastyrskaya K. MicroRNA MiR-199a-5p regulates smooth muscle cell proliferation and morphology by targeting WNT2 signaling pathway. J Biol Chem. 2015;290(11): 7067–7086. doi: 10.1074/jbc.M114.618694 [DOI] [PMC free article] [PubMed] [Google Scholar]
39.Taganov KD, Boldin MP, Chang KJ, Baltimore D. NF-κB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. PNAS. 2006;103(33): 12481–12486. [DOI] [PMC free article] [PubMed] [Google Scholar]
40.Herter EK, Xu Landén N. Non-coding RNAs: new players in skin wound healing. Adv Wound Care. 2017;6(3):93–107. doi: 10.1089/wound.2016.0711 [DOI] [PMC free article] [PubMed] [Google Scholar]
41.Tili E, Michaille JJ, Cimino A, Costinean S, Dumitru CD, Adair B, et al. Modulation of miR-155 and miR-125b levels following lipopolysaccharide/TNF-α stimulation and their possible roles in regulating the response to endotoxin shock. J Immun. 2007;179(8):5082–5089. [DOI] [PubMed] [Google Scholar]
42.Veith AP, Henderson K, Spencer A, Sligar AD, Baker AB. Therapeutic strategies for enhancing angiogenesis in wound healing. Adv Drug Deliv Rev. 2019;146: 97–125. doi: 10.1016/j.addr.2018.09.010 [DOI] [PMC free article] [PubMed] [Google Scholar]
43.Chan YC, Khanna S, Roy S, Sen CK. miR-200b targets Ets-1 and is down-regulated by hypoxia to induce angiogenic response of endothelial cells. J Biol Chem. 2011;286(3): 2047–2056. doi: 10.1074/jbc.M110.158790 [DOI] [PMC free article] [PubMed] [Google Scholar]
44.Safer JD. Thyroid hormone and wound healing. J Thyroid Res. 2013;2013: 124538. doi: 10.1155/2013/124538 [DOI] [PMC free article] [PubMed] [Google Scholar]
45.Liu X, Zheng N, Shi YN, Yuan J, Li L. Thyroid hormone induced angiogenesis through the integrin αvβ3/protein kinase D/histone deacetylase 5 signaling pathway. J Mol Endocrinol. 2014;52(3): 245–254. [DOI] [PubMed] [Google Scholar]
46.Losner J, Courtemanche K, Whited JL. A cross-species analysis of systemic mediators of repair and complex tissue regeneration. Npj Regen Med. 2012;6(1): 1–11. [DOI] [PMC free article] [PubMed] [Google Scholar]
47.Hong WX, Hu MS, Esquivel M, Liang GY, Rennert RC, McArdle A, et al. The role of hypoxia-inducible factor in wound healing. Adv Wound Care. 2014;3(5): 390–399. doi: 10.1089/wound.2013.0520 [DOI] [PMC free article] [PubMed] [Google Scholar]
48.Xu J, Liu X, Zhao F, Zhang Y, Wang Z. HIF1α overexpression enhances diabetic wound closure in high glucose and low oxygen conditions by promoting adipose-derived stem cell paracrine function and survival. Stem Cell Res Ther. 2020;11(1): 1–13. [DOI] [PMC free article] [PubMed] [Google Scholar]
49.Rajendran NK, Kumar SSD, Houreld NN, Abrahamse H. Understanding the perspectives of forkhead transcription factors in delayed wound healing. J Cell Commun Signal. 2019;13(2): 151–162. doi: 10.1007/s12079-018-0484-0 [DOI] [PMC free article] [PubMed] [Google Scholar]
50.Walton KL, Johnson KE, Harrison CA. Targeting TGF-β mediated SMAD signaling for the prevention of fibrosis. Front Pharmacol. 2017;8: 461. [DOI] [PMC free article] [PubMed] [Google Scholar]
51.Penn JW, Grobbelaar AO, Rolfe KJ. The role of the TGF-β family in wound healing, burns and scarring: a review. Int J Burn Trauma. 2012;2(1): 18. [PMC free article] [PubMed] [Google Scholar]
52.Liarte S, Bernabé-García Á, Nicolás FJ. Role of TGF-β in skin chronic wounds: a keratinocyte perspective. Cells. 2020;9(2): 306. [DOI] [PMC free article] [PubMed] [Google Scholar]
53.Zhang T, Wang XF, Wang ZC, Lou D, Fang QQ, Hu YY, et al. Current potential therapeutic strategies targeting the TGF-β/Smad signaling pathway to attenuate keloid and hypertrophic scar formation. Biomed Pharmacother. 2020;129: 110287. [DOI] [PubMed] [Google Scholar]
54.Jozic I, Vukelic S, Stojadinovic O, Liang L, Ramirez HA, Pastar I, et al. Stress signals, mediated by membranous glucocorticoid receptor, activate PLC/PKC/GSK-3β/β-catenin pathway to inhibit wound closure. J Invest Dermatol. 2017;137(5): 1144–1154. [DOI] [PMC free article] [PubMed] [Google Scholar]
55.Slominski AT, Zmijewski MA. Glucocorticoids inhibit wound healing: novel mechanism of action. J Invest Dermatol. 2017;137(5): 1012–1014. doi: 10.1016/j.jid.2017.01.024 [DOI] [PMC free article] [PubMed] [Google Scholar]
56.Ackers I, Malgor R. Interrelationship of canonical and non-canonical Wnt signalling pathways in chronic metabolic diseases. Diab Vasc Dis Res. 2018;15(1): 3–13. doi: 10.1177/1479164117738442 [DOI] [PMC free article] [PubMed] [Google Scholar]
57.Shi J, Barakat M, Chen D, Chen L. Bicellular tight junctions and wound healing. Int J Mol Sci. 2018;19(2): 3862. doi: 10.3390/ijms19123862 [DOI] [PMC free article] [PubMed] [Google Scholar]
58.Matsubayashi Y, Coulson-Gilmer C, Millard TH. Endocytosis-dependent coordination of multiple actin regulators is required for wound healing. Int J Cell Biol. 2015;210(3): 419–433. [DOI] [PMC free article] [PubMed] [Google Scholar]
59.Dabelsteen S, Pallesen EM, Marinova IN, Nielsen MI, Adamopoulou M, Rømer TB, et al. Essential functions of glycans in human epithelia dissected by a CRISPR-Cas9-engineered human organotypic skin model. Dev Cell. 2020;54(5): 669–684. doi: 10.1016/j.devcel.2020.06.039 [DOI] [PMC free article] [PubMed] [Google Scholar]
60.Jin SP, Chung JH. Inhibition of N-glycosylation by tunicamycin attenuates cell–cell adhesion via impaired desmosome formation in normal human epidermal keratinocytes. Biosci Rep. 2018;38(6): BSR20171641. doi: 10.1042/BSR20171641 [DOI] [PMC free article] [PubMed] [Google Scholar]
61.Sabbah DA, Hajjo R, Sweidan K. Review on epidermal growth factor receptor (EGFR) structure, signaling pathways, interactions, and recent updates of EGFR inhibitors. Curr Top Med Chem. 2020;20(10). doi: 10.2174/1568026620666200303123102 [DOI] [PubMed] [Google Scholar]
62.Kazanietz MG, Barrio-Real L, Casado-Medrano V, Baker MJ, Lopez-Haber C. The P-Rex1/Rac signaling pathway as a point of convergence for HER/ErbB receptor and GPCR responses. Small GTPases. 2018;94(2): 297–303. doi: 10.1080/21541248.2016.1221273 [DOI] [PMC free article] [PubMed] [Google Scholar]
63.Zhang L, Xu P, Wang X, Zhang M, Yan Y, Chen Y, et al. Activin B regulates adipose-derived mesenchymal stem cells to promote skin wound healing via activation of the MAPK signaling pathway. Int J Biochem. 2017;87: 69–76. doi: 10.1016/j.biocel.2017.04.004 [DOI] [PubMed] [Google Scholar]
64.Lee S, Kim MS, Jung SJ, Kim D, Park HJ, Cho D. ERK activating peptide, AES16-2M promotes wound healing through accelerating migration of keratinocytes. Sci Rep. 2018;8(1): 1–10. [DOI] [PMC free article] [PubMed] [Google Scholar]
65.Zhao N, Wang G, Long S, Hu M, Gao J, Ran X, et al. MicroRNA-34a deficiency leads to impaired wound closure by augmented inflammation in mice. Ann Transl Med. 2020;8(7). doi: 10.21037/atm.2020.03.161 [DOI] [PMC free article] [PubMed] [Google Scholar]
66.Wang T, Feng Y, Sun H, Zhang L, Hao L, Shi C, et al. miR-21 regulates skin wound healing by targeting multiple aspects of the healing process. Am J Pathol. 2012;181(6):1911–1920. doi: 10.1016/j.ajpath.2012.08.022 [DOI] [PubMed] [Google Scholar]
67.van Solingen C, Araldi E, Chamorro‐Jorganes A, Fernández‐Hernando C, Suárez Y. Improved repair of dermal wounds in mice lacking micro RNA‐155. (2014). J Cell Mol Med. 2014;18(6): 1104–1112. [DOI] [PMC free article] [PubMed] [Google Scholar]
68.Bleul T, Zhuang X, Hildebrand A, Lange C, Böhringer D, Schlunck G, et al. Different innate immune responses in BALB/c and C57BL/6 strains following corneal transplantation. J Innate Immun. 2021;13(1): 49–59. doi: 10.1159/000509716 [DOI] [PMC free article] [PubMed] [Google Scholar]

PLoS One. doi: 10.1371/journal.pone.0281913.r001

Decision Letter 0

Andrea Caporali

23 Dec 2021

PONE-D-21-30551MicroRNAs expressed during wound healing and their associated pathways: a systematic review and bioinformatics analysisPLOS ONE

Dear Dr. Azevedo,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Feb 06 2022 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Andrea Caporali, PhD

Academic Editor

PLOS ONE

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. Please report your search date in Search strategy section.

3. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match.

When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section.

4. Thank you for stating the following in the Acknowledgments Section of your manuscript:

The authors thank the financial support from the Brazilian National Council for Scientific and Technological Development (CNPq).

Please note that funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form.

Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows:

The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Please include your amended statements within your cover letter; we will change the online submission form on your behalf.

5. Please amend the manuscript submission data (via Edit Submission) to include author ROBERTA GIORGI SILVEIRA, FERNANDA NEDEL and RAFAEL GUERRA LUND.

6. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Partly

**********

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: I Don't Know

Reviewer #3: Yes

**********

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

**********

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: Yes

Reviewer #3: No

**********

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The authors present a timely and interesting analysis of miRNAs associated with wound healing in mouse skin. As the work is focused on physiological healing in healthy mice, perhaps the title needs to reflect this: MicroRNAs expressed during normal wound healing...

The manuscript needs editing for English language usage.

RESULTS

It is not entirely clear how the authors went from 2,370 articles with titles and abstracts to 915 articles for full text reading. If this this based on >1,400 articles either not being in English or not freely available, this needs to be made clear.

The authors state that day 3 was analysed by only one of the selected articles (page 10, line 188,189). But there are at least 6 studies in Table 2 that have a day 3 collection time: Aunin et al 2017, Chan et al 2012a, 2012b, Shi et al 2018, Wang et al 2019; Zhao et al 2020. So the authors need to remove or clarify that comment about day 3 analysis.

Looking at the day 1 and day 5 pathways for miRNA with increased expression on day 1 and day 5 (Tables 3 and 5), it might be useful to show the overlap between the pathways (e.g. using a Venn diagram). This would shed light on pathways that feature at both times points and delineate them from those that are limited to day 1 or day 5. Same argument goes for data in Tables 4 and 6, for miRNAs with reduced expression.

There were 273 miRNAs for day 1 and 327 for day 5 (p21, line 196ff). Conceivably all the 273 day 1 miRNAs featured in the 327 at day 5, but it would be good to make this clear or else generate a Venn diagram showing the overlap between the two datasets.

DISCUSSION

While the comments link miRNAs and targets to wound healing, there needs to be deeper consideration of the relationships between the miRNAs and the transcripts. Consider ErbB and MAPK signalling (p31,lines 338 ff). What makes these pathways noteworthy is not just that there are 18 and 39 target genes, respectively (Table 6) but rather the fact that they feature in the union pathways for miRNAs with DECREASED expression – that is, the targets themselves are likely to be upregulated to support cell proliferation given that the expression of their miRNA regulators is reduced. The same logic applies to most of the pathways considered.

The above also raises further questions: should additional/supplementary Results tables be generated showing the names of the target genes for the “interesting” pathways – “interesting” pathways being the ones considered in the Discussion?

In a similar vein, as presented, we cannot tell which miRNAs are associated with a given pathway. I am not sure if there is a way that allows this to be done easily but that seems to be the missing link between Table 2 and Tables 3-6. If not for all the targets, such insight might at least be relevant for those targets at the overlap/intersection of the day 1 and day 5 data that I mentioned in the Results.

Minor

The Simoes et al data in particular (in Table 2) have multiple entries that are not mouse miRNAs (i.e. not prefixed with mmu). This needs some explanation.

The papers in Tables 1 and 2, cited with author names, need the corresponding reference number from the bibliography for ease of cross-referencing.

Tables 3-5 have some pathways in bold. It is not clear what the bold font is supposed to signify, if anything.

Page 21,line 204: Table 2 rather than Table 4?

I could see Figure titles but the manuscript does not seem to have included figure legends.

Reviewer #2: Dear Author

1. MicroRNAs expressed during wound healing and their associated pathways: a systematic review and bioinformatics analysis require English corrections.

2. author has to include the miRs inhibition in wound healing and explain it in separate analysis

Reviewer #3: The systematic review by Azevedo et al. reported numerous miRNAs to regulate different phases of wound healing, in particular the inflammation and proliferation phases. The outcome of this review maybe used to guide preclinical and clinical studies on the role of miRNA in wound healing. In my view, the manuscript cannot be published in its present form.

Major comments:

1. Numerous typographical and grammatical errors have been observed. The English, formulation of certain sentences, and paragraphs need huge improvement to provide readers better understandability of the text.

2. Improvise abstract to provide gist of the manuscript.

3. Although the authors mentioned to include all articles involving in vivo animal studies, but only mice studies were included in table 1. Were there no studies using rats or rabbit?

4. Additionally, some points that need further clarification:

- In table 1, difference between back skin and dorsal skin?

- Sources of miRNAs, were the miRNAs extracted from peripheral blood or wound tissues?

5. It would be beneficial if the authors include these in the discussion:

- Comparison between miRNA signatures between day 1 and day 5.

- Relate the miRNA signatures or union pathways to the inflammatory and proliferation phases of wound healing.

**********

6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Kehinde Ross

Reviewer #2: No

Reviewer #3: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Attachment

Submitted filename: Azevedo et al PLOS ONE.docx

Click here for additional data file.^{(20.5KB, docx)}

Attachment

Submitted filename: PONE-D-21-30551 review report.pdf

Click here for additional data file.^{(119KB, pdf)}

PLoS One. 2023 Apr 13;18(4):e0281913. doi: 10.1371/journal.pone.0281913.r002

Author response to Decision Letter 0

15 Jun 2022

Journal Requirements:

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

Authors’ response:

We have modified the level of our manuscript titles and subtitles, and also, we added title to each to the figures, as required by the PLOS ONE. Thank you for your suggestions.

2. Please report your search date in Search strategy section.

Authors’ response:

Thank you very much for your suggestion. We added our search date in Search strategy section.

3. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match.

When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section.

Authors’ response:

We corrected the mistake in the ‘Funding Information’ and ‘Financial Disclosure’. Thank you for letting us know this mistake.

4. Thank you for stating the following in the Acknowledgments Section of your manuscript:

The authors thank the financial support from the Brazilian National Council for Scientific and Technological Development (CNPq).

Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows:

The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Please include your amended statements within your cover letter; we will change the online submission form on your behalf.

Authors’ response:

Thank you very much for your suggestion. We corrected the Acknowledgement section of our manuscript. The ‘Funding Statement’ information are already correct.

5. Please amend the manuscript submission data (via Edit Submission) to include author ROBERTA GIORGI SILVEIRA, FERNANDA NEDEL and RAFAEL GUERRA LUND.

Authors’ response:

The other authors’ names have been included. Thank you for letting us know this mistake.

Authors’ response:

The subtitles have been included. Thank you for letting us know this mistake.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Partly

Author’s comment:

Thank you for your comments. Our study was conducted rigorously with adequate controls and experimental design.

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: I Don't Know

Reviewer #3: Yes

Author’s comment:

The statistical analysis of our study has been performed adequately to support their conclusion. Thank you very much for your comments.

3. Have the authors made all data underlying the findings in their manuscript fully available?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Author’s comment:

Thank you very much for all your comments.

4. Is the manuscript presented in an intelligible fashion and written in standard English?

Reviewer #1: No

Reviewer #2: Yes

Reviewer #3: No

Author’s comment:

Thank you very much for all your comments. Our study has been corrected by an English teacher and the grammatical errors has been corrected.

5. Review Comments to the Author

Reviewer #1:

- The manuscript needs editing for English language usage.

Our study has been corrected by an English teacher and the grammatical errors has been corrected.

- RESULTS

Thank you very much for your comment. The explanation of why the articles went from 2370 to 915 are explained in the following lines: 156 and 157. After removing the duplicates, 2370 articles remained for the title and abstract screening, resulting in 915 articles.

Thank you very much for this comment. Actually, the sentence is poorly structured, because the day 3 was analyzed by more the one study. But, just in one we can see results that compare the up and down regulation. We really appreciate this comment. The authors had already corrected the error in the article.

- DISCUSSION

Thank you very much for this comment. The authors have already reformulated in the article.

Yes, the interesting pathways are those discussed in the discussion that have statistical significance.

I could see Figure titles but the manuscript does not seem to have included figure legends.

The authors already add figure legends.

Reviewer #2:

1. MicroRNAs expressed during wound healing and their associated pathways: a systematic review and bioinformatics analysis require English corrections.

Our study has been corrected by an English teacher and the grammatical errors has been corrected.

2. Author has to include the miRs inhibition in wound healing and explain it in separate analysis.

The inhibition of miRNAs in wound healing was included in the tables.

Major comments:

Thank you very much for this comment. We reviewed our study with the help of a professional English teacher and corrected any perceived errors.

2. Although the authors mentioned to include all articles involving in vivo animal studies, but only mice studies were included in table 1. Were there no studies using rats or rabbit?

No, none of the studies performed the analyzes in rats or rabbit.

3. Additionally, some points that need further clarification:

- In table 1, difference between back skin and dorsal skin?

According to the figures provided in the selected studies, there is no difference in “back skin” and “dorsal skin”.

- Sources of miRNAs, were the miRNAs extracted from peripheral blood or wound tissues?

The miRNAs were extracted from wound tissues.

4. It would be beneficial if the authors include these in the discussion:

- Comparison between miRNA signatures between day 1 and day 5.

- Relate the miRNA signatures or union pathways to the inflammatory and proliferation phases of wound healing.

Thank you very much for all this comments. The suggestions were accepted.

5. . PLOS authors have the option to publish the peer review history of their article. If published, this will include your full peer review and any attached files.

Yes.

PLoS One. doi: 10.1371/journal.pone.0281913.r003

Decision Letter 1

Andrea Caporali

18 Jul 2022

PONE-D-21-30551R1MicroRNAs expressed during wound healing and their associated pathways: a systematic review and bioinformatics analysisPLOS ONE

Dear Dr. Azevedo,

Please submit your revised manuscript by Sep 01 2022 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

We look forward to receiving your revised manuscript.

Kind regards,

Andrea Caporali, PhD

Section Editor

PLOS ONE

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: (No Response)

Reviewer #3: (No Response)

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

Reviewer #3: Partly

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

Reviewer #1: Yes

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

Reviewer #1: No

Reviewer #3: No

**********

6. Review Comments to the Author

Reviewer #1: While the authors have made some improvements, the work still needs substantial English language revision in my view.

Results: Perhaps "After exclusion of 1455 articles based on titles and abstract screening, 915 articles remained for the analysis" would make the point clearer.

Discussion: need for deeper considueration of the relationships between the miRNAs and their targets appears not to have been addressed.

Minor points raised appear not to be been addressed.

Reviewer #3: After comparing the original submission and the revised manuscript, the authors only made minimal changes to the subheadings and acknowledgement.

The manuscript still contain numerous typographical and grammatical errors, which the authors need to improve in order to provide readers better understandability of the text.

Additionally, the abstract needs to revise to provide the gist of the review.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Kehinde Ross

Reviewer #3: No

**********

PLoS One. 2023 Apr 13;18(4):e0281913. doi: 10.1371/journal.pone.0281913.r004

Author response to Decision Letter 1

21 Nov 2022

The authors received no specific funding for this work.

PLoS One. doi: 10.1371/journal.pone.0281913.r005

Decision Letter 2

Andrea Caporali

25 Nov 2022

PONE-D-21-30551R2MicroRNAs expressed during wound healing and their associated pathways: a systematic review and bioinformatics analysisPLOS ONE

Dear Dr.Azevedo,

Please submit your revised manuscript by Jan 09 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

We look forward to receiving your revised manuscript.

Kind regards,

Andrea Caporali, PhD

Section Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Additional Editor Comments:

Please consider minor comments from reviewer 1 and perform the language editing of the text.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #1: (No Response)

Reviewer #3: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Partly

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #3: N/A

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

Reviewer #1: Yes

Reviewer #3: No

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

Reviewer #1: No

Reviewer #3: Yes

**********

6. Review Comments to the Author

Reviewer #1: "However, the relationship between 191 the down and up expression on the day 3 was analyzed by only one of the selected articles". I can see two articles with both up and down regulated miRNA considered. Also, why some ters in tables 3-5 are bold remains unexplained. Issues with the language remain. I was going for "No recommendation" but that is not an option so either minor revision or reject but I will not look at this manuscript again.

Reviewer #3: (No Response)

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Kehinde Ross

Reviewer #3: No

**********

PLoS One. 2023 Apr 13;18(4):e0281913. doi: 10.1371/journal.pone.0281913.r006

Author response to Decision Letter 2

18 Jan 2023

January 4th, 2023

Dear Dr. Emily Chenette,

Editor-in-Chief of PLOS ONE

We would like to thank the reviewers for their consideration, which certainly contributed to improving the overall quality of the manuscript. We have considered all of them in the revised version, and re-submit our manuscript entitled “MicroRNAs expressed during wound healing and their associated pathways: a systematic review and bioinformatics analysis” for your further consideration.

We have prepared point-by-point responses to the review’s comments, and these are indicated below in red font. Also, the changes made in the text were tracked.

We hope that all these changes meet the requirements of the Journal and make the manuscript acceptable for publication in PLOS ONE. Thank you for considering the revised version of our manuscript.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #1: (No Response)

Reviewer #3: All comments have been addressed

Authors comments:

Thank you for your consideration. We really attempt to adequately address all reviewers' comments in a previous review round.

2. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Partly

Reviewer #3: Yes

Authors comments:

Thank you for your consideration. We already attempt to modify the original manuscript version with technically sound scientific research data that supports the conclusion. Besides, the systematic review methodology was conducted rigorously, with appropriate controls, replication, and sample sizes.

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #3: N/A

Authors comments:

Thank you for your consideration.

4. Have the authors made all data underlying the findings in their manuscript fully available?

Reviewer #1: Yes

Reviewer #3: No

Authors comments:

Thank you for your consideration. We attempt to make all data underlying the findings described in the manuscript fully available without restriction.

5. Is the manuscript presented in an intelligible fashion and written in standard English?

Reviewer #1: No

Reviewer #3: Yes

Authors comments:

Thank you for your consideration. In the current version of the manuscript, the language was corrected with the help of a professional English teacher.

6. Review Comments to the Author

Authors comments:

Thank you for your consideration. In fact, the day 3 was analyzed by more than one article. However, the alteration of the two possible patterns of expression (down or up) was verified in only one of the articles (Aunin et al., 2017). We have improved the sentence containing this information so that the explanation is clearer.

The bold terms in tables 3-5 are the signaling pathways related to the wound healing process. In the current version of the manuscript, we inserted this information before the tables, and also highlighted the signaling pathways related to the wound healing process in table 6, which was missing.

In the current version of the manuscript, the language was corrected with the assistance of a professional English teacher.

Reviewer #3: (No Response)

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

We have no problem to publish the peer review history of our article.

PLoS One. doi: 10.1371/journal.pone.0281913.r007

Decision Letter 3

Andrea Caporali

5 Feb 2023

MicroRNAs expressed during wound healing and their associated pathways: a systematic review and bioinformatics analysis

PONE-D-21-30551R3

Dear Dr. Azevedo,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Andrea Caporali, PhD

Section Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

PLoS One. doi: 10.1371/journal.pone.0281913.r008

Acceptance letter

Andrea Caporali

20 Feb 2023

PONE-D-21-30551R3

MicroRNAs expressed during normal wound healing and their associated pathways: a systematic review and bioinformatics analysis

Dear Dr. Azevedo:

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org.

If we can help with anything else, please email us at plosone@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr Andrea Caporali

Section Editor

PLOS ONE

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

S1 Checklist. PRISMA 2020 checklist.

(DOCX)

Click here for additional data file.^{(31.4KB, docx)}

S1 Table. Correlation of results regarding the expression pattern of miRNAs at different times of analysis.

(DOCX)

Click here for additional data file.^{(73.6KB, docx)}

Attachment

Submitted filename: Azevedo et al PLOS ONE.docx

Click here for additional data file.^{(20.5KB, docx)}

Attachment

Submitted filename: PONE-D-21-30551 review report.pdf

Click here for additional data file.^{(119KB, pdf)}

Data Availability Statement

All relevant data are within the paper.

[pone.0281913.ref001] 1.Rodrigues M, Kosaric N, Bonham CA, Gurtner GC. Wound healing: a cellular perspective. Physiol Rev. 2019;99: 665–706. doi: 10.1152/physrev.00067.2017 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref002] 2.Basu S, Shukla V. Complications of wound healing. In: Measurements in wound healing. Springer. 2012: 109–144. [Google Scholar]

[pone.0281913.ref003] 3.Wang PH, Huang BS, Horng HC, Yeh CC. Wound healing. J Chin Med Assoc. 2017;20: 1–8. [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref004] 4.Han G, Ceilley R. Chronic wound healing: a review of current management and treatments. Adv Ther. 2017;34: 599–610. doi: 10.1007/s12325-017-0478-y [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref005] 5.Yao Z, Niu J, Cheng B. Prevalence of chronic skin wounds and their risk factors in an inpatient hospital setting in northern China. Adv Wound Care. 2020;33: 1–10. [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref006] 6.Gupta S, Andersen C, Black J, de Leon J, Fife C, John CL, et al. Management of Chronic Wounds: Diagnosis, Preparation, Treatment, and Follow-up. Wounds. 2017;29: 19–36. [PubMed] [Google Scholar]

[pone.0281913.ref007] 7.Christopher AF, Kaur RP, Kaur G, Kaur A, Gupta V, Bansal P. MicroRNA therapeutics: Discovering novel targets and developing specific therapy. Perspect Clin Res. 2016;7: 68. doi: 10.4103/2229-3485.179431 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref008] 8.Rupaimoole R, Slack FJ. MicroRNA therapeutics: towards a new era for the management of cancer and other diseases. Nat Rev Drug Discov. 2017;16: 203–222. doi: 10.1038/nrd.2016.246 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref009] 9.Soliman AM, Das S, Abd Ghafar N, Teoh SL. Role of microRNA in proliferation phase of wound healing. Front genet. 2018;9: 38. doi: 10.3389/fgene.2018.00038 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref010] 10.Hanna J, Hossain GS, Kocerha J. The potential for microRNA therapeutics and clinical research. Front genet. 2019;10: 478. doi: 10.3389/fgene.2019.00478 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref011] 11.Catalanotto C, Cogoni C, Zardo G. MicroRNA in control of gene expression: an overview of nuclear functions. Int J Mol Sci. 2016;17: 1712. doi: 10.3390/ijms17101712 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref012] 12.Creugny A, Fender A, Pfeffer S. Regulation of primary micro RNA processing. FEBS Lett. 2018;592: 1980–1996. doi: 10.1002/1873-3468.13067 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref013] 13.Macgregor-Das AM, Das S. A microRNA’s journey to the center of the mitochondria. Am J Physiol Heart Circ Physiol. 2018;315: 206–215. [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref014] 14.Lu TX, Rothenberg ME. MicroRNA. J Allergy Clin Immunol. 2018;141: 1202–1207. doi: 10.1016/j.jaci.2017.08.034 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref015] 15.Piletič K, Kunej T. MicroRNA epigenetic signatures in human disease. Arch Toxicol. 2016;90: 2405–2419. doi: 10.1007/s00204-016-1815-7 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref016] 16.Yao Q, Chen Y, Zhou X. The roles of microRNAs in epigenetic regulation. Curr Opin Chem Biol. 2019;51: 11–17. doi: 10.1016/j.cbpa.2019.01.024 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref017] 17.Cui J, Zhou B, Ross SA, Zempleni J. Nutrition, microRNAs, and human health. Adv Nutr. 2017;8: 105–112. doi: 10.3945/an.116.013839 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref018] 18.Saliminejad K, Khorram Khorshid HR, Soleymani Fard S, Ghaffari SH. An overview of microRNAs: biology, functions, therapeutics, and analysis methods. J Cell Physiol. 2019;234: 5451–5465. doi: 10.1002/jcp.27486 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref019] 19.Aunin E, Broadley D, Ahmed MI, Mardaryev AN, Botchkareva NV. Exploring a role for regulatory miRNAs in wound healing during ageing: involvement of miR-200c in wound repair. Sci Rep. 2017;7: 1–10. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref020] 20.Chan YC, Roy S, Huang Y, Khanna S, Sem CK. The microRNA miR-199a-5p down-regulation switches on wound angiogenesis by derepressing the v-ets erythroblastosis virus E26 oncogene homolog 1-matrix metalloproteinase-1 pathway. J Biol Chem. 2012;287: 41032–41043. doi: 10.1074/jbc.M112.413294 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref021] 21.Etich J, Bergmeier V, Pitzler L, Brachvogel B. Identification of a reference gene for the quantification of mRNA and miRNA expression during skin wound healing. Connect Tissue Res. 2017;58: 196–207. doi: 10.1080/03008207.2016.1210606 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref022] 22.Shi J, Ma X, Su Y, Song Y, Tian Y, Yuan S, et al. MiR-31 mediates inflammatory signaling to promote re-epithelialization during skin wound healing. J Investig Dermatol. 2018; 138: 2253–2263. doi: 10.1016/j.jid.2018.03.1521 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref023] 23.Wang CR, Zhu HF, Zhu Y. Knockout of microRNA-155 ameliorates the Th17/Th9 immune response and promotes wound healing. Curr Med Sci. 2019;39: 954–964. doi: 10.1007/s11596-019-2128-x [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref024] 24.Long S, Zhao N, Ge L, Wang G, Ran X, Wang J, et al. MiR‐21 ameliorates age‐associated skin wound healing defects in mice. J Genet Med. 2018;20: 3022. doi: 10.1002/jgm.3022 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref025] 25.Boeker M, Vach W, Motschall E. Google Scholar as replacement for systematic literature searches: good relative recall and precision are not enough. BMC Med Res Methodol. 2013;13: 1–12. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref026] 26.Silveira RG, Ferrúa CP, do Amaral CC, Garcia TF, de Souza KB, Nedel, F. MicroRNAs expressed in neuronal differentiation and their associated pathways: systematic review and bioinformatics analysis. Brain Res Bull. 2020;157: 140–148. [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref027] 27.Wang W, Yang C, Wang X, yan Zhou L, Juan Lao G, Liu D, et al. MicroRNA-129 and-335 promote diabetic wound healing by inhibiting Sp1-mediated MMP-9 expression. Diabetes. 2018;67: 1627–1638. doi: 10.2337/db17-1238 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref028] 28.Jiang Z, Wei J, Yang W, Li W, Liu F, Yan X, et al. MicroRNA-26a inhibits wound healing through decreased keratinocytes migration by regulating ITGA5 through PI3K/AKT signaling pathway. Biosci Rep. 2020;40: BSR20201361. doi: 10.1042/BSR20201361 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref029] 29.Wu Y, Zhang K, Liu R, Zhang H, Chen D, Yu S, et al. MicroRNA-21-3p accelerates diabetic wound healing in mice by downregulating SPRY1. Aging. 2020;12: 15436. doi: 10.18632/aging.103610 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref030] 30.Zanoaga O, Braicu C, Chiroi P, Andreea N, Hajjar NA, Mărgărit S, et al. The Role of miR-155 in Nutrition: Modulating Cancer-Associated Inflammation. Nutrients. 2012;13(7): 2245. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref031] 31.Wolska-Gawron K, Bartosińska J, Rusek M, Kowal M, Raczkiewicz D, Krasowska D. Circulating miRNA-181b-5p, miRNA-223-3p, miRNA-210-3p, let 7i-5p, miRNA-21-5p and miRNA-29a-3p in patients with localized scleroderma as potential biomarkers. Sci Rep. 2020;10: 1–14. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref032] 32.Córdova-Rivas S, Fraire-Soto I, Mercado-Casas Torres A, Servín-González LS, Granados-López AJ, López-Hernández Y, et al. 5p and 3p strands of miR-34 family members have differential effects in cell proliferation, migration, and invasion in cervical cancer cells. Int J Mol Sci. 2019;20: 545. doi: 10.3390/ijms20030545 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref033] 33.Wu J, Li X, Li D, Ren X, Li Y, Herter EK, et al. MicroRNA-34 family enhances wound inflammation by targeting LGR4. J Investig Dermatol. 2020;140: 465–476. doi: 10.1016/j.jid.2019.07.694 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref034] 34.Childs DR, Murthy AS. Overview of wound healing and management. Surgical Clinics. 2017;97: 189–207. doi: 10.1016/j.suc.2016.08.013 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref035] 35.Chen L, Simões A, Chen Z, Zhao Y, Wu X, Dai Y, et al. Overexpression of the oral mucosa-specific microRNA-31 promotes skin wound closure. Int J Mol Sci. 2019;20: 3679. doi: 10.3390/ijms20153679 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref036] 36.Wang F, Gao Y, Yuan Y, Du R, Li P, Liu F, et al. MicroRNA-31 Can Positively Regulate the Proliferation, Differentiation and Migration of Keratinocytes. Biomed Hub. 2020;5(2): 1–12. doi: 10.1159/000508612 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref037] 37.Yan S, Xu Z, Lou F, Zhang L, Ke F, et al. NF-κB-induced microRNA-31 promotes epidermal hyperplasia by repressing protein phosphatase 6 in psoriasis. Nat Commun. 2015;6(1): 1–15. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref038] 38.Gheinani AH, Burkhard FC, Rehrauer H, Fournier CA, Monastyrskaya K. MicroRNA MiR-199a-5p regulates smooth muscle cell proliferation and morphology by targeting WNT2 signaling pathway. J Biol Chem. 2015;290(11): 7067–7086. doi: 10.1074/jbc.M114.618694 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref039] 39.Taganov KD, Boldin MP, Chang KJ, Baltimore D. NF-κB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. PNAS. 2006;103(33): 12481–12486. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref040] 40.Herter EK, Xu Landén N. Non-coding RNAs: new players in skin wound healing. Adv Wound Care. 2017;6(3):93–107. doi: 10.1089/wound.2016.0711 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref041] 41.Tili E, Michaille JJ, Cimino A, Costinean S, Dumitru CD, Adair B, et al. Modulation of miR-155 and miR-125b levels following lipopolysaccharide/TNF-α stimulation and their possible roles in regulating the response to endotoxin shock. J Immun. 2007;179(8):5082–5089. [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref042] 42.Veith AP, Henderson K, Spencer A, Sligar AD, Baker AB. Therapeutic strategies for enhancing angiogenesis in wound healing. Adv Drug Deliv Rev. 2019;146: 97–125. doi: 10.1016/j.addr.2018.09.010 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref043] 43.Chan YC, Khanna S, Roy S, Sen CK. miR-200b targets Ets-1 and is down-regulated by hypoxia to induce angiogenic response of endothelial cells. J Biol Chem. 2011;286(3): 2047–2056. doi: 10.1074/jbc.M110.158790 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref044] 44.Safer JD. Thyroid hormone and wound healing. J Thyroid Res. 2013;2013: 124538. doi: 10.1155/2013/124538 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref045] 45.Liu X, Zheng N, Shi YN, Yuan J, Li L. Thyroid hormone induced angiogenesis through the integrin αvβ3/protein kinase D/histone deacetylase 5 signaling pathway. J Mol Endocrinol. 2014;52(3): 245–254. [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref046] 46.Losner J, Courtemanche K, Whited JL. A cross-species analysis of systemic mediators of repair and complex tissue regeneration. Npj Regen Med. 2012;6(1): 1–11. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref047] 47.Hong WX, Hu MS, Esquivel M, Liang GY, Rennert RC, McArdle A, et al. The role of hypoxia-inducible factor in wound healing. Adv Wound Care. 2014;3(5): 390–399. doi: 10.1089/wound.2013.0520 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref048] 48.Xu J, Liu X, Zhao F, Zhang Y, Wang Z. HIF1α overexpression enhances diabetic wound closure in high glucose and low oxygen conditions by promoting adipose-derived stem cell paracrine function and survival. Stem Cell Res Ther. 2020;11(1): 1–13. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref049] 49.Rajendran NK, Kumar SSD, Houreld NN, Abrahamse H. Understanding the perspectives of forkhead transcription factors in delayed wound healing. J Cell Commun Signal. 2019;13(2): 151–162. doi: 10.1007/s12079-018-0484-0 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref050] 50.Walton KL, Johnson KE, Harrison CA. Targeting TGF-β mediated SMAD signaling for the prevention of fibrosis. Front Pharmacol. 2017;8: 461. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref051] 51.Penn JW, Grobbelaar AO, Rolfe KJ. The role of the TGF-β family in wound healing, burns and scarring: a review. Int J Burn Trauma. 2012;2(1): 18. [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref052] 52.Liarte S, Bernabé-García Á, Nicolás FJ. Role of TGF-β in skin chronic wounds: a keratinocyte perspective. Cells. 2020;9(2): 306. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref053] 53.Zhang T, Wang XF, Wang ZC, Lou D, Fang QQ, Hu YY, et al. Current potential therapeutic strategies targeting the TGF-β/Smad signaling pathway to attenuate keloid and hypertrophic scar formation. Biomed Pharmacother. 2020;129: 110287. [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref054] 54.Jozic I, Vukelic S, Stojadinovic O, Liang L, Ramirez HA, Pastar I, et al. Stress signals, mediated by membranous glucocorticoid receptor, activate PLC/PKC/GSK-3β/β-catenin pathway to inhibit wound closure. J Invest Dermatol. 2017;137(5): 1144–1154. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref055] 55.Slominski AT, Zmijewski MA. Glucocorticoids inhibit wound healing: novel mechanism of action. J Invest Dermatol. 2017;137(5): 1012–1014. doi: 10.1016/j.jid.2017.01.024 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref056] 56.Ackers I, Malgor R. Interrelationship of canonical and non-canonical Wnt signalling pathways in chronic metabolic diseases. Diab Vasc Dis Res. 2018;15(1): 3–13. doi: 10.1177/1479164117738442 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref057] 57.Shi J, Barakat M, Chen D, Chen L. Bicellular tight junctions and wound healing. Int J Mol Sci. 2018;19(2): 3862. doi: 10.3390/ijms19123862 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref058] 58.Matsubayashi Y, Coulson-Gilmer C, Millard TH. Endocytosis-dependent coordination of multiple actin regulators is required for wound healing. Int J Cell Biol. 2015;210(3): 419–433. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref059] 59.Dabelsteen S, Pallesen EM, Marinova IN, Nielsen MI, Adamopoulou M, Rømer TB, et al. Essential functions of glycans in human epithelia dissected by a CRISPR-Cas9-engineered human organotypic skin model. Dev Cell. 2020;54(5): 669–684. doi: 10.1016/j.devcel.2020.06.039 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref060] 60.Jin SP, Chung JH. Inhibition of N-glycosylation by tunicamycin attenuates cell–cell adhesion via impaired desmosome formation in normal human epidermal keratinocytes. Biosci Rep. 2018;38(6): BSR20171641. doi: 10.1042/BSR20171641 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref061] 61.Sabbah DA, Hajjo R, Sweidan K. Review on epidermal growth factor receptor (EGFR) structure, signaling pathways, interactions, and recent updates of EGFR inhibitors. Curr Top Med Chem. 2020;20(10). doi: 10.2174/1568026620666200303123102 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref062] 62.Kazanietz MG, Barrio-Real L, Casado-Medrano V, Baker MJ, Lopez-Haber C. The P-Rex1/Rac signaling pathway as a point of convergence for HER/ErbB receptor and GPCR responses. Small GTPases. 2018;94(2): 297–303. doi: 10.1080/21541248.2016.1221273 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref063] 63.Zhang L, Xu P, Wang X, Zhang M, Yan Y, Chen Y, et al. Activin B regulates adipose-derived mesenchymal stem cells to promote skin wound healing via activation of the MAPK signaling pathway. Int J Biochem. 2017;87: 69–76. doi: 10.1016/j.biocel.2017.04.004 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref064] 64.Lee S, Kim MS, Jung SJ, Kim D, Park HJ, Cho D. ERK activating peptide, AES16-2M promotes wound healing through accelerating migration of keratinocytes. Sci Rep. 2018;8(1): 1–10. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref065] 65.Zhao N, Wang G, Long S, Hu M, Gao J, Ran X, et al. MicroRNA-34a deficiency leads to impaired wound closure by augmented inflammation in mice. Ann Transl Med. 2020;8(7). doi: 10.21037/atm.2020.03.161 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref066] 66.Wang T, Feng Y, Sun H, Zhang L, Hao L, Shi C, et al. miR-21 regulates skin wound healing by targeting multiple aspects of the healing process. Am J Pathol. 2012;181(6):1911–1920. doi: 10.1016/j.ajpath.2012.08.022 [DOI] [PubMed] [Google Scholar]

[pone.0281913.ref067] 67.van Solingen C, Araldi E, Chamorro‐Jorganes A, Fernández‐Hernando C, Suárez Y. Improved repair of dermal wounds in mice lacking micro RNA‐155. (2014). J Cell Mol Med. 2014;18(6): 1104–1112. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0281913.ref068] 68.Bleul T, Zhuang X, Hildebrand A, Lange C, Böhringer D, Schlunck G, et al. Different innate immune responses in BALB/c and C57BL/6 strains following corneal transplantation. J Innate Immun. 2021;13(1): 49–59. doi: 10.1159/000509716 [DOI] [PMC free article] [PubMed] [Google Scholar]

PERMALINK

MicroRNAs expressed during normal wound healing and their associated pathways: A systematic review and bioinformatics analysis

Morgana Lüdtke Azevedo

Roberta Giorgi Silveira

Fernanda Nedel

Rafael Guerra Lund

Roles

Abstract

1. Introduction

2. Material and methods

2.1 Registration and review questions

2.2 Inclusion and exclusion criteria

2.3 Search strategy

2.4 Study selection

2.5 Data extraction

2.6 Bioinformatics analysis

2.7 Quality assessment

3. Results

Fig 1. Flow diagram of study selection.

Fig 2. Correlation between publication countries and years of selected articles.

Table 1. Correlation of the main methodological approaches found in the selected articles.

Table 3. Statistically significant union pathways refer to the miRNAs with a decreased expression on day 1.

Table 2. Statistically significant union pathways refer to the miRNAs with an increased expression on day 1.

Table 5. Statistically significant union pathways refer to the miRNAs with a decreased expression on day 5.

Table 4. Statistically significant union pathways refer to the miRNAs with an increased expression on day 5.

Fig 3. Analysis of the miRNAs expression.

Fig 4.

Fig 5. Analysis of risk of bias of the selected studies.

4. Discussion

5. Conclusion

Supporting information

Data Availability

Funding Statement

References

Decision Letter 0

Andrea Caporali

Roles

Author response to Decision Letter 0

Decision Letter 1

Andrea Caporali

Roles

Author response to Decision Letter 1

Decision Letter 2

Andrea Caporali

Roles

Author response to Decision Letter 2

Decision Letter 3

Andrea Caporali

Roles

Acceptance letter

Andrea Caporali

Roles

Associated Data

Supplementary Materials

Data Availability Statement

ACTIONS

PERMALINK

RESOURCES

Similar articles

Cited by other articles

Links to NCBI Databases