Abstract
Objective: This paper sets out to investigate the miR-205 and HMGB1 expressions in chronic periodontitis (PO) patients and their associations with the inflammatory factors. Methods: From February 2016 to May 2018, 68 PO patients treated in our hospital were recruited for the study and placed in a patient group (PG), and 60 healthy volunteers were also recruited and placed in a healthy group (HG). Serum samples were collected from both groups for the identification of miR-205 using qPCR, as well as to determine the HMGB1 and inflammatory factor (IL-1β, IL-6, TNF-α) expression levels using ELISA. The correlations of the miR-205 and HMGB1 expression levels with the periodontal clinical indicators and the inflammatory factors were analyzed using a correlation analysis. Results: In comparison with the HG expression, the serum miR-205 expression was lower, and the HMGB1 was elevated in PG (P < 0.05). An ROC curve analysis showed that the areas under the curve (AUCs) of the serum miR-205 and HMGB1 expressions in diagnosing PO were 0.936 and 0.955 respectively. However, the serum miR-205 expression in PG increased while the HMGB1 expression decreased post treatment (P < 0.05). A correlation analysis revealed an inverse association between the serum miR-205 expression levels and the periodontal clinical indicators [bleeding index (BI), probing depth (PD), plaque index (PLI), and attachment loss (AL)], and the inflammatory factors [interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α)], but there was a positive association between the HMGB1 expression level and these parameters. Conclusions: miR-205 and HMGB1 are closely related to the progression of PO, and may be candidate biomarkers for the diagnosis and treatment of PO.
Keywords: miR-205, HMGB1, chronic periodontitis, inflammatory factors
Introduction
Periodontitis (PO) is a type of chronic inflammation triggered by bacteria invading the gums and periodontal tissues [1]. The main clinical presentations are gingival inflammation and bleeding, alveolar bone absorption, attachment loss (AL), and periodontal pocket formation, which can eventually lead to tooth loss and seriously affect patients’ quality of life [2]. PO is a ubiquitous disease. According to the World Health Organization, 10-15% of the global population suffers from severe periodontitis [3]. It is not only considered as the primary cause of tooth loss in adults, it also increases the risk of multiple diseases such as cardiovascular disease, rheumatoid arthritis, adverse pregnancy outcomes, and cancer [4]. There is currently no valid method for treating the disease [5]. In view of this, it is paramount to better clarify the pathogenesis of periodontitis to find a better treatment.
miRs are highly conserved, endogenous short-chain non-coding RNAs, that are widely distributed in eukaryotic cells [6]. They can regulate physiological and pathological processes by blocking protein translation or inhibiting target gene expression by inducing mRNA degradation [7]. With our increasing understanding of miRs, they are not thought to play a key role in the development of a variety of inflammatory diseases. For example, miR-10a-5p is down-regulated in the synovia of rheumatoid arthritis patients, and through the targeted regulation of TBX5, it induces inflammatory changes in human synovial sarcoma cells [8]. Myeloid-derived miR-223 is able to regulate intestinal inflammation by inhibiting NLRP3 inflammasomes [9]. And miR-495 is up-regulated in osteoarthritis and can promote the development of osteoarthritis by regulating AKT1 [10]. miR-205, an important member of the miR family, has drawn much attention because of its participation in the development of many diseases [11-13]. Previous studies have reported that miR-205 is down-regulated in the gingival tissues of periodontitis patients [14]. The high mobility group protein 1 (HMGB1) is a protein type widely distributed in eukaryotic cells to regulate gene expression and transcription, and it is highly expressed in manifold inflammatory diseases and is considered to be an essential regulator involved in inflammatory progression [15-17]. Moreover, HMGB1 has been found to be highly expressed in periodontitis and facilitates its progression [18].
In this paper, we examined the serum miR-205 and HMGB1 expressions in patients with periodontitis, and analyze their significance in disease diagnosis and evaluation.
Materials and methods
Study participants
From August 2017 to August 2019, 86 patients with chronic periodontitis treated in the Affiliated Hospital of North Sichuan Medical College, recruited for this study, and placed in the patient group (PG), and 60 healthy controls who concurrently underwent healthy check-ups were also recruited for this study and placed in the healthy group (HG). Inclusion criteria for the PG: patients who met the diagnostic criteria of periodontitis [19], patients with complete clinical data, and patients with more than 16 teeth remaining in their mouths. Exclusion criteria of both groups: patients with an organic dysfunction of the heart, liver, kidneys, or lungs, patients with autoimmune diseases, patients who were pregnant, lactating, or menstruating, patients who had undergone prior orthodontic treatment, patients who had taken antibiotics or immunomodulatory drugs within the previous three months, patients who had taken drugs and/or who had undergone surgical treatment for periodontitis within the previous three months, and patients with incomplete clinical data. This study was approved by the Medical Ethics Committee of the Affiliated Hospital of North Sichuan Medical College, and all the participants were informed and signed the informed consent to participate.
Quantification of the clinical indicators and the serum sample collection
Oral and total dental examinations were performed by our professional oral surgeons in our hospital, and bleeding index (BI), periodontal probing depth (PD), plaque index (PLI), and attachment loss (AL) levels were recorded. The average values of each of the above clinical indicators were taken and compared between the two groups.
The participants were required to fast and were prohibited from smoking for two hours before their blood was drawn. Then 5 ml venous blood was drawn from the median cubital vein and centrifuged (4°C, 1500 × g, 10 min) for the serum collection and subsequent storage in a freezer at -80°C for later analysis.
qRT-PCR quantification
Total RNA was extracted using TRIzol kits and its purity was tested. Then, cDNA reverse transcription was carried out following the TaqMan™ Reverse kit (Invitrogen, USA) protocol, followed by PCR amplification via a PCR amplification kit (Takara Bio, Japan). The amplification system: 10 μL SYBR qPCR Mix, 0.8 μL each of upstream and downstream primers, 2 μL cDNA, 0.4 μL 50 × ROX reference dye, and water in a final volume of 20 μL. The reaction conditions were pre-denaturation at 95°C for 60 s, denaturation at 95°C for 30 s, and annealing at 60°C for 40 s, cycling 40 times. U6 was utilized as an internal reference, and the primers were designed by Shanghai Genechem Co., Ltd. miR-205 upstream sequence: 5’-GGCGTGAGGCTGAGGCTA-3’, downstream sequence: 5’-ATGGCTGAGCGAAATTGCGGAC-3’; U6 upstream sequence: 5’-CATCACCATCAGGAGAGTCG-3’, downstream sequence: 5’-TGACGCTTGCCCACAGCCTT-3’. Data processing was performed by 2-ΔΔct.
ELISA quantification
The serum inflammatory factors, including interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α), were quantified using ELISA. ELISA kits specific for IL-1β, IL-6, and TNF-α were purchased from Wuhan BOSTER Biotechnology Co., Ltd. with catalog numbers EK0412, EK0412, and EK0525 respectively.
Statistical processing
SPSS 21.0 (IBM Corp, Armonk, NY, USA) and GraphPad Prism 7 were used for the statistical analysis and figure rendering respectively. The count data were compared using either chi-square tests or Fisher’s exact tests. Independent sample t tests and paired t tests were used to carry the inter-group and intra-group comparisons, respectively. One-way analyses of variance were used for the comparisons among multiple groups of means, and Dunnett’s tests were used for the pairwise comparisons. The diagnostic value of miR-205 and HMGB1 in the diagnosis of periodontitis was evaluated using receiver operating characteristics (ROC) curves [the higher the area under the curve (AUC) value, the better the diagnostic significance]. Pearson method was used to analyze the correlations between variables. A p-value < 0.05 was considered significant for all the tests.
Results
Comparison of the general data
Significant differences were absent in the general patient data such as gender, age, weight, diet preference, residence, exercise habit, smoking history, and drinking history in the two groups (P > 0.05) (Table 1).
Table 1.
Comparison of the general information between the two series ([n (%)], x ± sd)
| Groups | Healthy group (n=60) | Patient group (n=68) | χ2/t | P |
|---|---|---|---|---|
| Gender | 1.729 | 0.189 | ||
| Male | 42 (70.00) | 40 (58.82) | ||
| Female | 18 (30.00) | 28 (41.28) | ||
| Age (years old) | 57.26±7.15 | 59.11±8.63 | 1.310 | 0.193 |
| Weight (KG) | 62.24±6.12 | 63.56±7.22 | 1.108 | 0.270 |
| Dietary preference | 0.728 | 0.394 | ||
| Light | 29 (48.33) | 38 (55.88) | ||
| Greasy | 31 (51.67) | 30 (44.12) | ||
| Residence | 0.343 | 0.558 | ||
| Urban | 34 (56.67) | 42 (61.76) | ||
| City | 26 (43.33) | 26 (38.24) | ||
| Do you have any exercise habits | 2.008 | 0.157 | ||
| Yes | 26 (43.33) | 38 (55.88) | ||
| No | 34 (56.67) | 30 (44.12) | ||
| History of smoking | 0.212 | 0.645 | ||
| Yes | 41 (68.33) | 49 (72.06) | ||
| No | 19 (31.67) | 19 (27.94) | ||
| History of drinking | 0.694 | 0.405 | ||
| Yes | 30 (50.00) | 29 (42.65) | ||
| No | 30 (50.00) | 39 (57.35) |
A comparison of the serum miR-205 and HMGB1 expressions
As indicated by the qRT-PCR and ELISA assays, PG presented notably lower, and the serum HMGB1 was higher than the HG (P < 0.001). A Pearson correlation analysis revealed an inverse correlation between the miR-205 and HMGB1 levels in the serum of PO patients (P < 0.001) (Figure 1).
Figure 1.
Comparison of the serum miR-205 and HMGB1 expressions in the two groups. A: Comparison of the serum miR-205 expressions in the two groups. B: Comparison of the serum HMGB1 expressions in the two groups. C: Correlation between the serum miR-205 and HMGB1 levels. Note: ***P < 0.001.
The diagnostic effect of the serum miR-205 and HMGB1 on periodontitis
The ROC curves revealed that the AUC of serum miR-205 in the diagnosis of periodontitis was 0.936, the sensitivity was 88.24%, and the specificity was 86.67%. The AUC, the sensitivity, and the specificity of the serum HMGB1 in diagnosing PO were 0.955, 89.71% and 95.00% respectively (Table 2; Figure 2).
Table 2.
The diagnostic value of the serum miR-205 and HMGB1 expression levels in periodontitis
| Groups | AUC | 95% CI | Standard error | Cut-off value | Sensitivity (%) | Specificity (%) |
|---|---|---|---|---|---|---|
| miR-205 | 0.936 | 0.896-0.975 | 0.020 | 0.811 | 88.24 | 86.67 |
| HMGB1 | 0.955 | 0.916-0.995 | 0.020 | 3.364 | 89.71 | 95.00 |
Figure 2.
The ROC curves of the serum miR-205 and HMGB1 levels in the diagnosis of periodontitis. The AUC of serum miR-205 in the diagnosis of periodontitis was 0.936, the sensitivity was 88.24%, and the specificity was 86.67%. The AUC, sensitivity, and specificity of serum HMGB1 in the diagnosis of periodontitis were 0.955, 89.71% and 95.00%, respectively.
Expression changes in the miR-205 and HMGB1 levels before and after the treatment
Comparing the expressions of miR-205 and HMGB1 before and after the periodontal basic treatment, it was found that miR-205 was increased while HMGB1 decreased post treatment (P < 0.001) (Figure 3).
Figure 3.
Changes in the serum miR-205 and HMGB1 expressions before and after the treatment. A: Changes in the serum miR-205 expressions before and after the treatment. B: Changes in the serum HMGB1 expressions before and after the treatment. Note: ***P < 0.001.
The correlations of the miR-205 and HMGB1 expressions with the periodontal clinical indicators
The results of periodontal clinical indexes of the patients were PD (5.45±0.78) mm, AL (4.59±0.68) mm, PLI (1.11±0.17), and GI (1.21±0.19). A Pearson correlation analysis showed that miR-205 is negatively related to PD, AL, PLI and GI, and HMGB1 is positively correlated with PD, AL, PLI, and GI (Figure 4).
Figure 4.
The correlation of miR-205 and HMGB1 with the periodontal clinical indicators. A-D: The correlation between serum miR-205 and the periodontal clinical indicators PD, AL, PLI, and GI. E-H: The correlation between serum HMGB1 and the periodontal clinical indicators PD, AL, PLI, and GI.
Comparison of the serum IL-1β, IL-6, and TNF-α levels in the two groups
ELISA indicated that the serum IL-1β, IL-6, and TNF-α levels were higher in the PG than in the HG (P < 0.001) (Figure 5).
Figure 5.
A comparison of the serum IL-1β, IL-6, and TNF-α levels in the two groups. A: A comparison of the serum IL-1β levels in the two groups. B: A comparison of the serum IL-6 levels in the two groups. C: A comparison of the serum TNF-α levels in the two groups. Note: *** P < 0.001.
The correlation of the miR-205 and HMGB1 levels with the serum inflammatory factor levels
A Pearson correlation analysis showed that miR-205 has an inverse association with the IL-1β, IL-6, and TNF-α levels (P < 0.001), and that HMGB1 has a positive correlation with the above inflammatory factor levels (P < 0.001) (Figure 6).
Figure 6.
The correlation of miR-205 and HMGB1 with the serum inflammatory factors. A-C: The correlation between serum miR-205 and the inflammatory factors IL-1β, IL-6, and TNF-α in the serum. D-F: The correlation between the serum HMGB1 levels and the inflammatory factors IL-1β, IL-6, and TNF-α in the serum.
Discussion
The investigation of the role of miRs in various diseases reveals that miRs are essential for the pathological development of periodontitis, which provides a new perspective and strategy for treating the disease [20,21]. HMGB1 is a highly-conserved DNA binding protein that interferes with many biological processes such as DNA transcription, gene recombination, DNA repair, and nucleosome formation [22]. In PO, HMGB1 is abnormally elevated and can be released in the extracellular mediating inflammatory response [23]. In this paper, it was found that PO patients presented with notably lower serum miR-205 expressions and higher serum HMGB1 expressions than the controls, which agrees with the findings of previous studies [14,24]. We also observed that serum miR-205 increased and HMGB1 decreased post treatment. Further, a negative association was identified between miR-205 and the periodontal clinical indicators PD, AL, PLI, and GI, while a positive correlation between HMGB1 and these periodontal clinical indicators was identified. These results suggest that miR-205 and HMGB1 are closely related to the occurrence of PO and may be potential targets for its treatment. Past clinical experience suggests that prevention should be the priority for PO, and the early diagnosis and observation of PO progression can provide important information for PO treatment [25]. However, the early clinical diagnosis of periodontitis is not satisfactory, so finding efficient diagnostic methods is also warranted [26]. Previous studies have found that miR and HMGB1 are potential biomarkers for the diagnosis of many diseases [27,28]. Subsequently, we analyzed the diagnostic effect of serum miR-205 and HMGB1 on periodontitis using ROC curves. The results demonstrated that the AUCs of serum miR-205 and HMGB1 in the diagnosis of PO were 0.936 and 0.955 respectively, suggesting that serum miR-205 and HMGB1 have the potential to serve as biomarkers for the diagnosis of PO.
The concentrations of the inflammatory factors such as IL-1β, IL-6, and TNF-α are reported to be significantly increased in periodontitis, a finding similar to the results of this study [29]. In the process of periodontitis, cytokines are closely related to inflammation, the immune response, and periodontal tissue destruction [30]. TNF-α is a common and crucial inflammatory factor in the development of PO, and it can promote the formation of osteoclasts, inhibit the transformation of periodontal ligament fibroblasts into osteoblasts, and the activity of osteoblasts, resulting in the destruction of periodontal connective tissue and alveolar bone tissue, thus limiting the repair of periodontal tissue [30,31]. IL-6 can facilitate the release of inflammatory mediators to aggravate the inflammatory reaction, inhibit the growth of periodontal ligament cells, and hinder the repair of periodontal ligaments, as well as induce the production of matrix metalloproteinase, ultimately leading to the degradation of bone matrix [32]. And IL-1β, produced mainly by monocytes, macrophages and neutrophils, is critical in inflammation and immunity [33]. To further understand the relationship between miR-205, HMGB1, and periodontitis, we analyzed the correlation between the serum miR-205, HMGB1 and inflammatory factor levels in PO patients. The results showed that miR-205 is negatively correlated with IL-1β, IL-6, and TNF-α, and HMGB1 is positively correlated with the above inflammatory factors. This suggests that miR-205 and HMGB1 may participate in PO progression by modulating the secretions of the cytokines such as IL-1β, IL-6, and TNF-α.
Finally, our Pearson correlation analysis revealed an inverse correlation between miR-205 and HMGB1. It is shown that miR-205 participates in the progression of sepsis, septicemia, and triple negative breast cancer through the targeted regulation of HMGB1 [13,34,35]. Therefore, miR-205 can also participate in the development of PO through the targeted regulation of HMGB1 expression. However, this study did not design a basic experiment for demonstrating this. So it is hoped that relevant experiments for verification will be carried out in subsequent studies.
To sum up, miR-205 and HMGB1 are closely related to the progression of periodontitis and may be candidate biomarkers for its diagnosis and treatment.
Disclosure of conflict of interest
None.
References
- 1.Hajishengallis G, Korostoff JM. Revisiting the Page & Schroeder model: the good, the bad and the unknowns in the periodontal host response 40 years later. Periodontol 2000. 2017;75:116–151. doi: 10.1111/prd.12181. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2.Papapanou PN, Sanz M, Buduneli N, Dietrich T, Feres M, Fine DH, Flemmig TF, Garcia R, Giannobile WV, Graziani F, Greenwell H, Herrera D, Kao RT, Kebschull M, Kinane DF, Kirkwood KL, Kocher T, Kornman KS, Kumar PS, Loos BG, Machtei E, Meng H, Mombelli A, Needleman I, Offenbacher S, Seymour GJ, Teles R, Tonetti MS. Periodontitis: consensus report of workgroup 2 of the 2017 World Workshop on the Classification of Periodontal and Peri-Implant Diseases and Conditions. J Clin Periodontol. 2018;45(Suppl 20):S162–S170. doi: 10.1111/jcpe.12946. [DOI] [PubMed] [Google Scholar]
- 3.Joshi D, Garg T, Goyal AK, Rath G. Advanced drug delivery approaches against periodontitis. Drug Deliv. 2016;23:363–377. doi: 10.3109/10717544.2014.935531. [DOI] [PubMed] [Google Scholar]
- 4.Hoare A, Soto C, Rojas-Celis V, Bravo D. Chronic inflammation as a link between periodontitis and carcinogenesis. Mediators Inflamm. 2019;2019:1029857. doi: 10.1155/2019/1029857. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.An JY, Quarles EK, Mekvanich S, Kang A, Liu A, Santos D, Miller RA, Rabinovitch PS, Cox TC, Kaeberlein M. Rapamycin treatment attenuates age-associated periodontitis in mice. Geroscience. 2017;39:457–463. doi: 10.1007/s11357-017-9994-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6.Li Q, Liang X, Wang Y, Meng X, Xu Y, Cai S, Wang Z, Liu J, Cai G. miR-139-5p inhibits the epithelial-mesenchymal transition and enhances the chemotherapeutic sensitivity of colorectal cancer cells by downregulating BCL2. Sci Rep. 2016;6:27157. doi: 10.1038/srep27157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7.Stark A, Bushati N, Jan CH, Kheradpour P, Hodges E, Brennecke J, Bartel DP, Cohen SM, Kellis M. A single Hox locus in Drosophila produces functional microRNAs from opposite DNA strands. Genes Dev. 2008;22:8–13. doi: 10.1101/gad.1613108. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8.Hussain N, Zhu W, Jiang C, Xu J, Wu X, Geng M, Hussain S, Cai Y, Xu K, Xu P, Han Y, Sun J, Meng L, Lu S. Down-regulation of miR-10a-5p in synoviocytes contributes to TBX5-controlled joint inflammation. J Cell Mol Med. 2018;22:241–250. doi: 10.1111/jcmm.13312. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9.Neudecker V, Haneklaus M, Jensen O, Khailova L, Masterson JC, Tye H, Biette K, Jedlicka P, Brodsky KS, Gerich ME, Mack M, Robertson AAB, Cooper MA, Furuta GT, Dinarello CA, O’Neill LA, Eltzschig HK, Masters SL, McNamee EN. Myeloid-derived miR-223 regulates intestinal inflammation via repression of the NLRP3 inflammasome. J Exp Med. 2017;214:1737–1752. doi: 10.1084/jem.20160462. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10.Zhao X, Wang T, Cai B, Wang X, Feng W, Han Y, Li D, Li S, Liu J. MicroRNA-495 enhances chondrocyte apoptosis, senescence and promotes the progression of osteoarthritis by targeting AKT1. Am J Transl Res. 2019;11:2232–2244. [PMC free article] [PubMed] [Google Scholar]
- 11.Eyking A, Reis H, Frank M, Gerken G, Schmid KW, Cario E. MiR-205 and miR-373 are associated with aggressive human mucinous colorectal cancer. PLoS One. 2016;11:e0156871. doi: 10.1371/journal.pone.0156871. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12.Zhou CF, Liu MJ, Wang W, Wu S, Huang YX, Chen GB, Liu LM, Peng DX, Wang XF, Cai XZ, Li XX, Feng WQ, Ma Y. miR-205-5p inhibits human endometriosis progression by targeting ANGPT2 in endometrial stromal cells. Stem Cell Res Ther. 2019;10:287. doi: 10.1186/s13287-019-1388-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13.Wang L, Kang FB, Wang J, Yang C, He DW. Downregulation of miR-205 contributes to epithelial-mesenchymal transition and invasion in triple-negative breast cancer by targeting HMGB1-RAGE signaling pathway. Anticancer Drugs. 2019;30:225–232. doi: 10.1097/CAD.0000000000000705. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14.Stoecklin-Wasmer C, Guarnieri P, Celenti R, Demmer RT, Kebschull M, Papapanou PN. MicroRNAs and their target genes in gingival tissues. J Dent Res. 2012;91:934–940. doi: 10.1177/0022034512456551. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Andersson U, Yang H, Harris H. Extracellular HMGB1 as a therapeutic target in inflammatory diseases. Expert Opin Ther Targets. 2018;22:263–277. doi: 10.1080/14728222.2018.1439924. [DOI] [PubMed] [Google Scholar]
- 16.Sun X, Zeng H, Wang Q, Yu Q, Wu J, Feng Y, Deng P, Zhang H. Glycyrrhizin ameliorates inflammatory pain by inhibiting microglial activation-mediated inflammatory response via blockage of the HMGB1-TLR4-NF-kB pathway. Exp Cell Res. 2018;369:112–119. doi: 10.1016/j.yexcr.2018.05.012. [DOI] [PubMed] [Google Scholar]
- 17.Patel V, Dial K, Wu J, Gauthier AG, Wu W, Lin M, Espey MG, Thomas DD, Ashby CR Jr, Mantell LL. Dietary antioxidants significantly attenuate hyperoxia-induced acute inflammatory lung injury by enhancing macrophage function via reducing the accumulation of airway HMGB1. Int J Mol Sci. 2020;21:977. doi: 10.3390/ijms21030977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18.Yoshihara-Hirata C, Yamashiro K, Yamamoto T, Aoyagi H, Ideguchi H, Kawamura M, Suzuki R, Ono M, Wake H, Nishibori M, Takashiba S. Anti-HMGB1 neutralizing antibody attenuates periodontal inflammation and bone resorption in a murine periodontitis model. Infect Immun. 2018;86:e00111–18. doi: 10.1128/IAI.00111-18. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19.Armitage GC. Periodontal diagnoses and classification of periodontal diseases. Periodontol 2000. 2004;34:9–21. doi: 10.1046/j.0906-6713.2002.003421.x. [DOI] [PubMed] [Google Scholar]
- 20.Wang L, Wu F, Song Y, Li X, Wu Q, Duan Y, Jin Z. Long noncoding RNA related to periodontitis interacts with miR-182 to upregulate osteogenic differentiation in periodontal mesenchymal stem cells of periodontitis patients. Cell Death Dis. 2016;7:e2327. doi: 10.1038/cddis.2016.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21.Lian J, Wu X, Liu Y, Qiu W, Zhu X, Wang X, Meng S, Valverde P, Steffensen B, Tu Q, Pan J, Chen J. Potential roles of miR-335-5p on pathogenesis of experimental periodontitis. J Periodontal Res. 2020;55:191–198. doi: 10.1111/jre.12701. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22.Deng M, Scott MJ, Fan J, Billiar TR. Location is the key to function: HMGB1 in sepsis and trauma-induced inflammation. J Leukoc Biol. 2019;106:161–169. doi: 10.1002/JLB.3MIR1218-497R. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 23.Lin YC, Wu CY, Chang LY, Chen CC, Chen HH, Lai YL, Hung SL. Levels of high-mobility group box-1 in gingival crevicular fluid in nonsmokers and smokers with chronic periodontitis. J Formos Med Assoc. 2017;116:933–939. doi: 10.1016/j.jfma.2017.01.006. [DOI] [PubMed] [Google Scholar]
- 24.Chapple ILC, Mealey BL, Van Dyke TE, Bartold PM, Dommisch H, Eickholz P, Geisinger ML, Genco RJ, Glogauer M, Goldstein M, Griffin TJ, Holmstrup P, Johnson GK, Kapila Y, Lang NP, Meyle J, Murakami S, Plemons J, Romito GA, Shapira L, Tatakis DN, Teughels W, Trombelli L, Walter C, Wimmer G, Xenoudi P, Yoshie H. Periodontal health and gingival diseases and conditions on an intact and a reduced periodontium: consensus report of workgroup 1 of the 2017 World Workshop on the classification of periodontal and peri-implant diseases and conditions. J Periodontol. 2018;89(Suppl 1):S74–S84. doi: 10.1002/JPER.17-0719. [DOI] [PubMed] [Google Scholar]
- 25.Patil PB, Patil BR. Saliva: a diagnostic biomarker of periodontal diseases. J Indian Soc Periodontol. 2011;15:310–317. doi: 10.4103/0972-124X.92560. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26.Mico-Martinez P, Garcia-Gimenez JL, Seco-Cervera M, Lopez-Roldan A, Alminana-Pastor PJ, Alpiste-Illueca F, Pallardo FV. miR-1226 detection in GCF as potential biomarker of chronic periodontitis: a pilot study. Med Oral Patol Oral Cir Bucal. 2018;23:e308–e314. doi: 10.4317/medoral.22329. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 27.Ghweil AA, Osman HA, Hassan MH, Sabry AM, Mahdy RE, Ahmed AR, Okasha A, Khodeary A, Ameen HH. Validity of serum amyloid A and HMGB1 as biomarkers for early diagnosis of gastric cancer. Cancer Manag Res. 2020;12:117–126. doi: 10.2147/CMAR.S207934. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 28.Xie P, Deng LX, Gong P, Ding Y, Tang XH. Expression of HMGB1 and HMGN2 in gingival tissues, GCF and PICF of periodontitis patients and peri-implantitis. Braz J Microbiol. 2011;42:1213–1219. doi: 10.1590/S1517-838220110003000047. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 29.Cardoso EM, Reis C, Manzanares-Cespedes MC. Chronic periodontitis, inflammatory cytokines, and interrelationship with other chronic diseases. Postgrad Med. 2018;130:98–104. doi: 10.1080/00325481.2018.1396876. [DOI] [PubMed] [Google Scholar]
- 30.Li X, Yang H, Zhang Z, Yan Z, Lv H, Zhang Y, Wu B. Concentrated growth factor exudate enhances the proliferation of human periodontal ligament cells in the presence of TNFalpha. Mol Med Rep. 2019;19:943–950. doi: 10.3892/mmr.2018.9714. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 31.Kim JH, Kim AR, Choi YH, Jang S, Woo GH, Cha JH, Bak EJ, Yoo YJ. Tumor necrosis factor-alpha antagonist diminishes osteocytic RANKL and sclerostin expression in diabetes rats with periodontitis. PLoS One. 2017;12:e0189702. doi: 10.1371/journal.pone.0189702. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 32.Toker H, Gorgun EP, Korkmaz EM. Analysis of IL-6, IL-10 and NF-kappaB gene polymorphisms in aggressive and chronic periodontitis. Cent Eur J Public Health. 2017;25:157–162. doi: 10.21101/cejph.a4656. [DOI] [PubMed] [Google Scholar]
- 33.Moeintaghavi A, Arab HR, Rahim Rezaee SA, Naderi H, Shiezadeh F, Sadeghi S, Anvari N. The effects of smoking on expression of IL-12 and IL-1beta in gingival tissues of patients with chronic periodontitis. Open Dent J. 2017;11:595–602. doi: 10.2174/1874210601711010595. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 34.Zhou W, Wang J, Li Z, Li J, Sang M. MicroRNA-2055b inhibits HMGB1 expression in LPS-induced sepsis. Int J Mol Med. 2016;38:312–318. doi: 10.3892/ijmm.2016.2613. [DOI] [PubMed] [Google Scholar]
- 35.Zhang Y, Xia F, Wu J, Yang AX, Zhang YY, Zhao H, Tao WY. MiR-205 influences renal injury in sepsis rats through HMGB1-PTEN signaling pathway. Eur Rev Med Pharmacol Sci. 2019;23:10950–10956. doi: 10.26355/eurrev_201912_19798. [DOI] [PubMed] [Google Scholar]