Abstract
Background:
Exosomes mediate tissue regeneration by promoting angiogenesis, fibroblast activity, and extracellular matrix remodeling. New Cellular Treatment Factor (NCTF), cellular treatment factor solutions containing amino acids, vitamins, nucleotides, coenzymes, minerals, and hyaluronic acid, are used in regenerative medicine, but their combined effects with human-derived exosomes on the oral mucosa remain unclear. The aim of this study was to evaluate the histological and immunohistochemical changes in normal hamster oral mucosa following treatment with an NCTF solution enriched with 2 distinct human exosome formulations at different time points.
Methods:
Sixty-three male hamsters were randomized into 3 groups: saline-injected control, NCTF with cell-derived exosomes (CellExosome), and NCTF solution with adipose stem cell–derived exosomes (ASCEs). Injections were administered into the buccal mucosa, and tissues were harvested at baseline, day 3, and day 7. Hematoxylin–eosin, Masson trichrome, and Van Gieson staining were used to assess mucosal structure and collagen remodeling, whereas angiogenesis was quantified using CD34 immunohistochemistry.
Results:
Both exosome-treated groups demonstrated progressive increases in fibroblast proliferation, collagen organization, and microvessel density from day 3 to day 7 compared with controls. The NCTF + CellExosome group consistently exhibited the most pronounced effects, with significantly greater angiogenesis and extracellular matrix remodeling across all time points (P < 0.05).
Conclusions:
In a hamster model, exosome-enriched biorevitalization solutions enhanced angiogenesis and collagen remodeling in normal oral mucosa in a time-dependent manner. The NCTF + CellExosome formulation demonstrated superior regenerative potential, supporting its potential translational relevance for future regenerative and aesthetic research.
Takeaways
Question: Does combining a biorevitalization solution (New Cellular Treatment Factor [NCTF]) with human-derived exosomes enhance oral mucosal regeneration compared with controls and other exosome formulations?
Findings: Both NCTF + CellExosome and NCTF + ASCEs (adipose stem cell–derived exosomes) improved epithelial regeneration, collagen remodeling, and angiogenesis. However, the NCTF + CellExosome group consistently produced the most pronounced histological and immunohistochemical improvements.
Meaning: Biorevitalization enriched with cell-derived exosomes represents a promising strategy to optimize mucosal regeneration, with potential applications in both regenerative and aesthetic medicine.
INTRODUCTION
The oral mucosa is a unique tissue with remarkable repair and regeneration capacity. Unlike skin, it heals faster with less scarring due to differences in extracellular matrix, angiogenesis, and immune modulation. These features make it a key target in regenerative medicine and aesthetic surgery. Therapies that enhance fibroblast activity, collagen remodeling, and vascularization in the oral mucosa are increasingly relevant.1–5
Among emerging tools in regenerative medicine, extracellular vesicles—particularly small extracellular vesicles or exosomes—have gained significant attention. These nanosized vesicles (30–150 nm), secreted by most cell types, transport bioactive molecules such as proteins, lipids, and nucleic acids. They play key roles in intercellular communication, regulating angiogenesis, fibroblast proliferation, and extracellular matrix remodeling. Studies in dermatology and tissue engineering show that human cell–derived exosomes stimulate collagen synthesis, increase vascular density, and accelerate repair. Their use in aesthetic medicine is rapidly expanding, especially as adjuncts to biorevitalization protocols.6–9
New Cellular Treatment Factor (NCTF), a biorevitalizing polycomponent solution, is a cornerstone of regenerative and aesthetic practice. These injectable formulations contain amino acids, vitamins, nucleotides, minerals, coenzymes, and hyaluronic acid, providing substrates for cellular metabolism, collagen synthesis, and antioxidant defense. Hyaluronic acid supports hydration and tissue structure, whereas cofactors and amino acids enable enzymatic processes essential for repair. Widely used in mesotherapy and facial rejuvenation, these formulations improve dermal thickness, hydration, and extracellular matrix integrity. Despite clinical popularity, the molecular and histological mechanisms underlying their effects remain incompletely understood.10,11
The combination of exosomes with polycomponent biorevitalization solutions is an attractive concept that may yield synergistic benefits. Exosomes provide bioactive signals that activate regenerative pathways, whereas the biorevitalization solution supplies the substrates and cofactors required for tissue remodeling. Together, they may enhance fibroblast activity, promote angiogenesis, and optimize extracellular matrix architecture. Studies suggest that exosomes can enhance cell proliferation and migration in wound healing.12,13 Nevertheless, there is a scarcity of in vivo studies addressing the combined effects of these 2 approaches, particularly on intact mucosal tissues.
The oral mucosa is an excellent model for evaluating regenerative interventions. Its structural similarity to skin and unique wound-healing properties allow translational insights relevant to oral and cutaneous applications.14 Although many studies on exosomes and biorevitalization have focused on injured or aged tissues, their effects on normal mucosa remain poorly studied. Understanding the baseline impact on healthy tissue is essential to assess safety, potential overstimulation, and the capacity to precondition tissues before surgery or aesthetic procedures.
To date, no comprehensive in vivo study has assessed how different human-derived exosome formulations, combined with a biorevitalization cocktail, affect intact oral mucosa. It remains unclear whether such protocols induce angiogenic or extracellular matrix remodeling responses in healthy tissue, or whether different exosome preparations produce distinct outcomes. Addressing these questions is essential, as exosome-enriched therapies are increasingly adopted in aesthetic and regenerative medicine despite an incomplete understanding of their effects on normal tissues.
By analyzing the angiogenic and matrix remodeling effects of exosome-enriched biorevitalization on intact oral mucosa, this study provided new insights into potential applications in regenerative and aesthetic medicine. The findings may guide future clinical strategies to enhance oral and skin tissue vitality and support the safe, effective translation of these therapies into practice.
MATERIALS AND METHODS
ETHICAL APPROVAL
All experimental procedures were conducted in accordance with the international guidelines for the care and use of laboratory animals and followed the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines. Ethical approval was obtained from an institutional animal care and use committee.
Using a hamster model, we evaluated the histological and immunohistochemical changes induced in normal oral mucosa by a biorevitalization solution enriched with either cell-derived exosomes or adipose stem cell–derived exosomes (ASCEs), both human exosomes. Saline-injected mucosa served as a control. Tissues were examined at baseline, day 3, and day 7 to capture both early and short-term responses. Histological assessments included hematoxylin–eosin, Masson trichrome, and Van Gieson staining to evaluate mucosal architecture and collagen remodeling, whereas angiogenesis was quantified through CD34 immunohistochemistry. We hypothesized that both exosome-enriched formulations would promote fibroblast proliferation, angiogenesis, and extracellular matrix remodeling, with ASCEs exerting superior effects compared with cell-derived exosomes.
Animals
A total of 63 healthy male golden Syrian hamsters (Mesocricetus auratus) were used in this study. Animals were aged 10–12 weeks and weighed 90–120 g at baseline. All animals were obtained from the accredited Animal House Facility at Cairo University. Before enrollment, animals underwent veterinary screening to exclude signs of systemic illness, mucosal lesions, or parasitic infestation. Only clinically healthy animals were included.
Hamsters were housed in individually ventilated polycarbonate cages (40 × 25 × 20 cm) with up to 5 animals per cage to avoid stress. Wood-shaving bedding was changed twice weekly, and cages included paper rolls and shelters to promote natural behavior. Housing was maintained at 22 ± 2°C, 50%–60% humidity, with a 12-hour light/dark cycle. Animals received standard rodent chow (Purina Rodent Chow) with 18% protein, 5% fat, and balanced vitamins and minerals, and filtered water ad libitum. Food intake, water consumption, and body weights were monitored regularly to ensure uniform health across groups.
To minimize experimental variability, all animals were acclimatized for at least 7 days before the beginning of interventions. During this period, hamsters were handled daily by the same researchers to reduce handling-related stress. Animals were randomly allocated into experimental groups using a computer-generated randomization list, and cage allocation was blinded to the investigator performing histological and immunohistochemical analyses.
Study Design, Grouping, Randomization, and Blinding
This was a prospective, randomized, controlled, blinded in vivo study with 2 experimental arms and 1 control group. Animals were assessed at baseline (day 0), day 3, and day 7 postinjection.
Group I (control): Injection of sterile saline solution into the buccal mucosa.
Group II (cell-derived exosome group): Injection of a biorevitalization polycomponent solution (NCTF 135 HA; Fillmed Laboratories, Paris, France) combined with human cell–derived exosomes (CellExosome, Black Label; Abio Materials Co., Ltd., Seoul, South Korea).
Group III (ASCE group): Injection of the same biorevitalization solution (NCTF) combined with human ASCEs (ExoCoBio Inc., Seoul, South Korea).
The NCTF and exosome formulations were self-funded and purchased personally by the authors, with no external financial support, institutional funding, or donations involved. Each group consisted of 21 animals and was subdivided into 3 standardized subgroups (n = 7) corresponding to follow-up periods: baseline, day 3, and day 7.
Randomization was performed using a computer-generated sequence (block size = 6). Allocation concealment was ensured via sequentially numbered, opaque, sealed envelopes opened only at the time of injection:
Performance blinding: The injector was aware only of coded vials (A, B, or C) and not their contents.
Detection blinding: Histotechnologists, image analysts, and pathologists were blinded to group and time point; slides were anonymized.
Data blinding: Statistical analyses were performed on coded datasets, with unblinding only after primary analyses were completed.
Anesthesia and Injection Protocol
Animals were anesthetized with a mixture of ketamine (0.03 mL; Sigma-Aldrich, St. Louis, MO) and xylazine (0.02 mL; Bayer, Leverkusen, Germany) via intraperitoneal injection. Depth of anesthesia was confirmed by loss of the pedal withdrawal reflex.
For experimental groups, equal volumes of NCTF and the corresponding exosomes were mixed immediately before injection. Group II received 0.05 mL of NCTF combined with 0.05 mL of CellExosome, and group III received 0.05 mL of NCTF combined with 0.05 mL of ASCEs. Each mixture (0.1 mL) was injected submucosally into the buccal mucosa at a depth of 2 mm using a 30G insulin syringe.
Control animals received 0.1 mL sterile saline (0.9% NaCl; Fresenius Kabi, Bad Homburg, Germany) under identical conditions. All injections were performed by the same investigator at standardized anatomical sites.
Tissue Harvesting
Tissue harvesting was standardized to ensure reproducibility. Full-thickness buccal mucosa samples (6 × 6 mm) were collected from the left side, 5 mm posterior to the oral commissure. Orientation was maintained by marking the superior edge and applying colored ink to the margins. Specimens were pinned epithelium side up and fixed in 10% neutral-buffered formalin for 24 hours. Blocks were embedded with the epithelium perpendicular to the cutting plane for subsequent histological and immunohistochemical analysis.
Tissue Processing, Histological Evaluation, and Immunohistochemistry
After fixation, tissue samples were processed using standardized dehydration, clearing, and paraffin-embedding protocols. Serial 5-µm-thick sections were cut from the central region of each specimen and mounted on poly-l-lysine–coated slides to ensure consistency. Histological evaluation was performed using hematoxylin and eosin to assess general tissue architecture, epithelial integrity, and cellular morphology. Collagen deposition and organization were evaluated using Masson trichrome, in which collagen fibers appear blue, and Van Gieson staining, in which collagen fibers appear red. All staining procedures were performed in a single batch per staining type to minimize variability, and slides were analyzed in a blinded manner by 2 independent histopathologists.
Immunohistochemical analysis was performed to evaluate angiogenesis using CD34 as an endothelial cell marker. Sections were deparaffinized, rehydrated, and subjected to antigen retrieval in citrate buffer (pH 6.0). Endogenous peroxidase activity was blocked with 3% hydrogen peroxide, followed by incubation with a protein-blocking solution to reduce nonspecific binding. Primary antibodies against CD34 were applied at optimized dilutions and incubated overnight at 4°C. After washing, sections were incubated with biotinylated secondary antibodies and streptavidin–horseradish peroxidase conjugate. Diaminobenzidine was used as the chromogen, and sections were counterstained with hematoxylin. Quantitative analysis of CD34-positive microvessels was performed at 100× magnification, with 3 representative fields analyzed per section to calculate mean microvessel density. All analyses were performed in a blinded manner to avoid detection bias.
To ensure standardization and reproducibility, all injections, tissue harvesting, sectioning, and staining procedures were performed by the same investigators following uniform protocols. Tissue orientation and sectioning planes were preserved, and interobserver reliability was assessed for histological and immunohistochemical scoring, with discrepancies resolved by consensus. This approach minimized variability and ensured unbiased, reliable assessment across all groups and time points.
Statistical Analysis
All quantitative data are presented as mean ± SD. Normality of the data distribution was assessed using the Shapiro–Wilk test. For comparisons among the 3 experimental groups at each follow-up time point, 1-way analysis of variance was performed, followed by the Tukey post hoc test for multiple comparisons. Nonnormally distributed data were analyzed using the Kruskal–Wallis test with Dunn multiple comparison post hoc test. Statistical significance was set at a P value less than 0.05 for all analyses.
All analyses were conducted using coded datasets to maintain blinding. Investigators performing the statistical tests were unaware of group allocations or time points, and unblinding occurred only after completion of all analyses. Graphical representations of the data, including bar charts with error bars and line graphs for temporal changes, were generated using standard software to ensure reproducibility and clarity. This standardized statistical approach minimized bias and allowed for robust, reliable comparisons across groups and time points.
RESULTS
All 63 hamsters tolerated the experimental procedures without complications, and no local or systemic adverse effects were observed. Body weight, food intake, and general behavior remained stable across all groups, confirming the safety of the interventions.
Histological Assessment (Hematoxylin and Eosin Staining)
At baseline, buccal mucosa across all groups displayed normal architecture with a stratified squamous epithelium, intact basal and spinous layers, and a lamina propria with sparse fibroblasts and minimal vascularity. By day 3, both experimental groups (NCTF + CellExosome and NCTF + ASCEs) showed early epithelial thickening, slight basal cell proliferation, and increased fibroblast presence compared with the control group, which remained histologically unchanged. By day 7, groups II and III exhibited pronounced epithelial hyperplasia, stratification, and keratinization, accompanied by dense fibroblast infiltration. Group II (NCTF + CellExosome) demonstrated greater epithelial height and fibroblast density than group III, suggesting a stronger regenerative response. In contrast, the control group retained baseline epithelial structure with minimal cellular changes (Figs. 1–5).
Fig. 1.
Hematoxylin and eosin–stained section of hamster buccal mucosa from the NCTF + CellExosome group at baseline (day 0), showing normal epithelial architecture.
Fig. 5.
Hematoxylin and eosin–stained section of hamster buccal mucosa from the NCTF + ASCE group at day 7 postinjection, showing further epithelial hyperplasia, less pronounced than in the NCTF + CellExosome group.
Fig. 2.
Hematoxylin and eosin–stained section of hamster buccal mucosa from the NCTF + CellExosome group at day 3 postinjection, showing early epithelial thickening and increased fibroblast presence.
Fig. 3.
Hematoxylin and eosin–stained section of hamster buccal mucosa from the NCTF + CellExosome group at day 7 postinjection, showing marked epithelial hyperplasia and dense fibroblast infiltration.
Fig. 4.
Hematoxylin and eosin–stained section of hamster buccal mucosa from the NCTF + ASCE group at day 3 postinjection, showing moderate epithelial thickening and fibroblast activation.
Collagen Evaluation (Masson Trichrome and Van Gieson Staining)
Collagen deposition and organization were assessed at all time points. At day 3, experimental groups displayed early collagen fiber deposition with moderate alignment compared with controls, which showed loose and irregular collagen bundles. By day 7, both experimental groups demonstrated dense, thick collagen bundles with improved orientation, confirming enhanced connective tissue remodeling. Notably, group II showed more uniform collagen alignment and higher staining intensity than group III, indicating superior matrix organization. Control specimens showed faint, loosely organized collagen fibers with negligible structural change throughout the study (Figs. 6–10).
Fig. 6.
Masson trichrome–stained section of hamster buccal mucosa from the NCTF + CellExosome group at baseline (day 0), showing sparse collagen fibers.
Fig. 10.
Masson trichrome–stained section of hamster buccal mucosa from the NCTF + ASCE group at day 7 postinjection, showing increased collagen organization with lower intensity than the NCTF + CellExosome group.
Fig. 7.
Masson trichrome–stained section of hamster buccal mucosa from the NCTF + CellExosome group at day 3 postinjection, showing early collagen deposition and alignment.
Fig. 8.
Masson trichrome–stained section of hamster buccal mucosa from the NCTF + CellExosome group at day 7 postinjection, showing dense and well-organized collagen bundles.
Fig. 9.
Masson trichrome–stained section of hamster buccal mucosa from the NCTF + ASCE group at day 3 postinjection, showing moderate collagen deposition.
Angiogenesis Assessment (CD34 Immunohistochemistry)
CD34 immunostaining revealed microvessel density changes across groups and time points. Baseline staining was comparable across all groups. By day 3, experimental groups exhibited a moderate increase in CD34-positive microvessels, with group II showing slightly higher counts than group III. By day 7, both experimental groups demonstrated marked increases in microvessel density compared with controls, reflecting enhanced angiogenesis. Group II consistently showed the highest vessel density and more complex capillary networks. Control animals maintained low microvessel density and minimal vascular proliferation throughout the study (Figs. 11–16).
Fig. 11.
CD34 immunohistochemical staining of hamster buccal mucosa from the NCTF + CellExosome group at baseline (day 0), showing low microvessel density.
Fig. 16.
CD34 immunohistochemical staining of hamster buccal mucosa from the NCTF + ASCE group at day 7 postinjection, showing increased angiogenesis with lower vessel density than the NCTF + CellExosome group.
Fig. 12.
CD34 immunohistochemical staining of hamster buccal mucosa from the NCTF + CellExosome group at day 3 postinjection, showing increased CD34-positive microvessels.
Fig. 13.
CD34 immunohistochemical staining of hamster buccal mucosa from the NCTF + CellExosome group at day 7 postinjection, showing marked angiogenesis and complex capillary networks.
Fig. 14.
CD34 immunohistochemical staining of hamster buccal mucosa from the NCTF + ASCE group at baseline (day 0), showing low microvessel density.
Fig. 15.
CD34 immunohistochemical staining of hamster buccal mucosa from the NCTF + ASCE group at day 3 postinjection, showing moderate angiogenic response.
Temporal and Quantitative Trends
Across all parameters—epithelial thickness, fibroblast density, collagen deposition, and microvessel density—the effects were progressive over time. Early changes were detectable at day 3, with more robust responses by day 7. Quantitative evaluation confirmed statistically significant differences between experimental groups and controls at both postinjection time points, with the NCTF + CellExosome group consistently demonstrating superior tissue remodeling outcomes (Tables 1–4).
Table 1.
Epithelial Thickness (µm) of Hamster Buccal Mucosa in Control, NCTF + CellExosome, and NCTF + ASCE Groups at Baseline (Day 0), Day 3, and Day 7 Postinjection
| Group | Baseline (Day 0) | Day 3 | Day 7 | P (Versus Control) |
|---|---|---|---|---|
| Control | 45.2 ± 3.1 | 46.0 ± 2.8 | 46.5 ± 3.0 | — |
| NCTF + ASCE | 45.4 ± 2.9 | 53.8 ± 3.5 | 61.2 ± 4.1 | <0.001* |
| NCTF + CellExosome | 45.1 ± 3.0 | 56.5 ± 3.2 | 66.8 ± 4.5 | <0.001* |
Values are expressed as mean ± SD (n = 7 per subgroup). Statistical analysis was performed using 1-way analysis of variance followed by the Tukey post hoc test. Among experimental groups, NCTF + CellExosome demonstrated consistently greater epithelial thickening, particularly by day 7.
P < 0.05 compared with control at the corresponding time point.
Table 4.
Microvessel Density (CD34 Immunohistochemistry): Microvessel Density (CD34-Positive Vessels Per Field at 100× Magnification) in Hamster Buccal Mucosa of all Experimental Groups at Baseline, Day 3, and Day 7
| Group | Baseline (Day 0) | Day 3 | Day 7 | P (Versus Control) |
|---|---|---|---|---|
| Control | 4.5 ± 1.0 | 4.7 ± 1.1 | 5.0 ± 1.2 | — |
| NCTF + ASCE | 4.6 ± 0.9 | 7.8 ± 1.2 | 12.3 ± 1.5 | <0.001* |
| NCTF + CellExosome | 4.5 ± 1.0 | 8.5 ± 1.1 | 14.1 ± 1.6 | <0.001* |
Values are expressed as mean ± SD (n = 7 per subgroup). Statistical significance was determined using 1-way analysis of variance followed by the Tukey post hoc test.
P < 0.05 versus control at the same time point.
Table 2.
Fibroblast Density: Fibroblast Density (Cells Per Field at 100× Magnification) in Hamster Buccal Mucosa Across Experimental Groups and Time Points
| Group | Baseline (Day 0) | Day 3 | Day 7 | P (Versus Control) |
|---|---|---|---|---|
| Control | 12.5 ± 2.0 | 13.1 ± 2.3 | 13.5 ± 2.1 | — |
| NCTF + ASCE | 12.8 ± 2.1 | 21.4 ± 2.5 | 28.7 ± 3.0 | <0.001* |
| NCTF + CellExosome | 12.7 ± 2.0 | 23.2 ± 2.8 | 32.1 ± 3.2 | <0.001* |
Values are presented as mean ± SD (n = 7 per subgroup). One-way analysis of variance with Tukey post hoc test was used for statistical comparison.
P < 0.05 versus control at the same follow-up interval.
Table 3.
Collagen Deposition Score: Semi-quantitative Assessment of Collagen Deposition in Hamster Buccal Mucosa Using Masson Trichrome and Van Gieson Staining
| Group | Baseline (Day 0) | Day 3 | Day 7 | P (Versus Control) |
|---|---|---|---|---|
| Control | 0.8 ± 0.3 | 0.9 ± 0.2 | 1.0 ± 0.3 | — |
| NCTF + ASCE | 0.9 ± 0.2 | 2.2 ± 0.3 | 3.0 ± 0.4 | <0.001* |
| NCTF + CellExosome | 0.8 ± 0.3 | 2.5 ± 0.3 | 3.5 ± 0.4 | <0.001* |
Scores ranged from 0 (absent) to 4 (dense). Data are expressed as mean ± SD (n = 7 per subgroup). Differences between groups were analyzed using 1-way analysis of variance with Tukey post hoc test.
P < 0.05 compared with control at the corresponding time point.
Overall Observations
The combined interventions of NCTF with human-derived exosomes significantly enhanced epithelial regeneration, connective tissue remodeling, and angiogenesis in intact hamster oral mucosa. The effect was more pronounced in the NCTF + CellExosome group compared with the NCTF + ASCE group, indicating a potential advantage of cell-derived exosomes for tissue regeneration. Control animals maintained baseline mucosal architecture, confirming that the observed changes were specifically induced by the experimental interventions.
DISCUSSION
This study demonstrated that combining a biorevitalization solution (NCTF) with human-derived exosomes enhances epithelial regeneration, collagen remodeling, and angiogenesis in normal hamster oral mucosa. The NCTF + CellExosome group consistently outperformed the NCTF + ASCE group across histological and immunohistochemical parameters, indicating superior regenerative capacity of cell-derived exosomes. These findings provide insights into the effects of exosome-enriched biorevitalization on intact oral mucosa and highlight the translational potential of this approach in regenerative and aesthetic medicine.
Our results align with evidence that exosomes are key mediators of tissue regeneration by transferring proteins, microRNAs, and growth factor–related molecules that regulate proliferation, extracellular matrix deposition, and angiogenesis. Studies in cutaneous and dermal models show that human exosomes enhance fibroblast proliferation, upregulate type I collagen synthesis, and stimulate endothelial tube formation, accelerating wound closure and improving scar quality.15–20 This study extended these findings to the oral mucosa and highlighted the added benefit of combining exosomes with a polycomponent biorevitalization solution. As no NCTF-only group was included, the outcomes reflect the synergistic effects of NCTF and exosomes, consistent with clinical application.
Cell-derived exosomes showed greater efficacy than ASCEs, particularly in promoting epithelial hyperplasia, collagen alignment, and microvessel density. This differential response may stem from variations in cellular origin and bioactive cargo. CellExosomes, enriched in growth factors, matrix-modulating proteins, and angiogenic microRNAs, likely enhance fibroblast activity and vascular sprouting more effectively than ASCEs. Their synergy with NCTF amino acids, vitamins, and cofactors may further amplify metabolism and collagen synthesis, producing superior tissue remodeling. Future proteomic and transcriptomic analyses are needed to clarify the molecular mechanisms underlying these distinct effects.
From a translational standpoint, the superior outcomes achieved with NCTF combined with CellExosome have important implications. In oral and maxillofacial surgery, mucosal preconditioning with such formulations may enhance healing after grafting, implant placement, or reconstruction. In aesthetic medicine, where biorevitalization is used to improve dermal hydration and elasticity, incorporating cell-derived exosomes could represent a next-generation approach for more durable rejuvenation. The pronounced angiogenic response observed here is significant, as increased vascularization supports faster healing and sustained tissue vitality. These findings warrant further investigation of exosome-enriched biorevitalization for optimizing oral tissue quality and regenerative outcomes.
The strengths of this study include a standardized animal model, rigorous randomization and blinding, and complementary histological and immunohistochemical analyses to assess epithelial, connective tissue, and vascular changes. Examination of intact rather than injured mucosa demonstrated that exosome-enriched biorevitalization can induce regenerative responses even without overt injury. This supports the concept of tissue “preconditioning,” in which improving baseline tissue quality enhances resilience and promotes optimal healing after surgical or aesthetic procedures.
Potential clinical indications for mucosal biorevitalization with NCTF and exosomes include conditions with mucosal thinning, reduced vascularity, or atrophy, such as postradiation mucositis, chronic irritation, or presurgical tissue conditioning. Although derived from healthy human stem cells and generally promoting physiological regeneration, the mitogenic and pro-angiogenic properties of exosomes warrant caution in pathological contexts. As studies show, exosomes may either inhibit or promote malignant activity depending on their source and cargo; therefore, use in tissues with dysplastic or malignant potential should be avoided until safety is fully established.
Several limitations of this study should be acknowledged. First, as an animal study, the extent to which hamster mucosal responses mirror human tissues remains uncertain. Second, the 7-day follow-up captured only short-term changes; longer term effects on collagen maturation, vascular stability, and safety require evaluation. Third, although histological and immunohistochemical analyses provided valuable insights, molecular assessments such as gene expression and exosome cargo profiling were not performed. Only 2 exosome preparations were tested; future studies should examine additional formulations, doses, and repeated treatments. Finally, as the study focused on combined NCTF–exosome effects, an NCTF-only group was excluded. Subsequent research should include NCTF- and exosome-only controls to clarify their individual and synergistic contributions.
CONCLUSIONS
NCTF combined with human-derived exosomes enhanced epithelial regeneration, collagen remodeling, and angiogenesis in normal oral mucosa. The NCTF + CellExosome group showed the strongest effects, indicating superior regenerative potential to ASCEs. These results support the translational value of cell-derived exosome–enriched biorevitalization, though further studies are needed to confirm long-term safety and efficacy.
DISCLOSURE
The authors have no financial interest to declare in relation to the content of this article.
ETHICAL APPROVAL
All experimental procedures were conducted in accordance with the international guidelines for the care and use of laboratory animals and followed the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines. Ethical approval was obtained from an Institutional Animal Care and Use Committee.
Footnotes
Published online 8 April 2026.
Disclosure statements are at the end of this article, following the correspondence information.
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