UNIVERSA MEDICINA
pISSN: 1907-3062 / eISSN: 2407-2230 Univ Med 2025.
44:131-140
DOI: https://doi.
org/10.
18051/UnivMed.
ORIGINAL ARTICLE
Hypoxia-preconditioned mesenchymal stem cells attenuate proinflammatory cytokines in collagen loss animal model Yeni Fristiani1* .
Agung Putra2,3,4 .
Titiek Sumarawati4 .
Eko Setiawan4,5 .
Sugeng Ibrahim2,6 , and Dodik Tugasworo Pramukarso7 Student of Postgraduate Biomedical Science.
Universitas Islam Sultan Agung.
Semarang.
Indonesia Stem Cell and Cancer Research (SCCR) Laboratory.
Semarang.
Indonesia Department of Doctoral Biomedical Science.
Faculty of Medicine.
Universitas Islam Sultan Agung.
Semarang.
Indonesia Department of Postgraduate Biomedical Science.
Universitas Islam Sultan Agung.
Semarang.
Indonesia Department of Surgery.
Faculty of Medicine.
Universitas Islam Sultan Agung.
Semarang.
Indonesia Faculty of Medicine.
Soegijapranata Catholic University.
Semarang.
Indonesia Neurology Department.
Faculty of Medicine.
Diponegoro University/Dr.
Kariadi Hospital.
Semarang.
Indonesia * Correspondence Author:
Yeninila77@gmail.
Date of first submission.
January 28, 2025 Date of final revised submission.
June 2, 2025 Date of acceptance.
July 1, 2025 Cite this article as: Fristiani Y.
Putra A.
Sumarawati T.
Setiawan E.
Ibrahim S.
Pramukarso DT.
Hypoxia-preconditioned mesenchymal stem cells attenuate proinflammatory cytokines in collagen loss animal model.
Univ Med 2025.
44:131-140
ABSTRACT
BACKGROUND
Repeated ultraviolet-B (UVB) exposure induces significant collagen degradation, primarily through overproduction of reactive oxygen species, which subsequently drives an inflammatory cascade.
Hypoxiapreconditioned mesenchymal stem cells (H-MSC.
constitute a promising therapeutic approach to counteract collagen loss by modulating inflammatory pathways.
This study aimed to evaluate the potential of H-MSCs in regulating NF-B p65 and IL-1 expression in a collagen loss rat model, highlighting their therapeutic efficacy.
METHODS
Twenty-five healthy male Wistar rats were randomly assigned to five groups: K1 .
ealthy control.
K2 .
ollagen los.
K3 .
ollagen loss hyaluronic aci.
K4 .
ollagen loss 2.
5 y 10AA H-MSC.
, and K5 .
ollagen loss 5 y 10AA H-MSC.
Collagen loss was induced by UVB radiation .
eak wavelength: 302 n.
for 2 weeks.
mRNA expression of NF-B p65 was quantified by qRT-PCR, while IL-1 levels were assessed using ELISA.
The rats were maintained for 14 days before being sacrificed, to allow the H-MSCs to exert their therapeutic Data analysis was by One-way ANOVA with TukeyAos post-hoc test.
RESULTS
The administration of H-MSCs significantly reduced IL-1 levels in groups K4 .
14A63.
76 pg/mL) and K5 .
80A123.
82 pg/mL) compared to group K2 .
93A205.
80 pg/mL) .
<0.
, with group K5 showing the most substantial reduction.
Moreover.
H-MSC injection in groups K4 and K5 effectively reduced NF-B p65 expression levels .
13A0.
50 a.
72A0.
22 a.
, respectivel.
, compared to group K2 .
47A0.
50 a.
<0.
, with group K5 providing optimum inhibition.
Copyright@Author.
- https://univmed.
org/ejurnal/index.
php/medicina/article/view/1702 Fadrian F.
Decroli E.
Ahmad A, et al
CONCLUSION
This study demonstrated that H-MSCs effectively attenuate UVB-induced inflammation and modulate key inflammatory pathways.
Keywords: Collagen loss.
H-MSCs, p65.
IL-1, rats INTRODUCTION Repeated exposure to ultraviolet-B (UVB) radiation leads to significant collagen loss, characterized by reduction in epidermal thickness, degradation of elastic fibers, and increased skin wrinkling and dryness.
This damage compromises the extracellular matrix (ECM), primarily composed of structural proteins such as collagen and elastin, which are crucial for maintaining skin .
UVB-induced collagen degradation is closely associated with the overproduction of reactive oxygen species (ROS), which trigger proinflammatory pathways, including the activation of nuclear factor kappa B (NF-B) subunit p65 and elevation of interleukin-1 (IL.
This inflammatory cascade upregulates degradative enzymes that accelerate collagen degradation while suppressing collagen .
Current therapies, such as retinol and hyaluronic acid (HA), and clinical procedures such as laser resurfacing and botulinum toxin injections, present significant side effects and .
Emerging evidence suggests that hypoxia-preconditioned mesenchymal stem cells (H-MSC.
may offer a novel therapeutic strategy for UVB-induced collagen loss.
Hypoxia-preconditioned mesenchymal stem cells (H-MSC.
offer promising therapeutic potential in mitigating collagen loss by modulating inflammatory pathways.
Sourced from tissues such as the umbilical cord, bone marrow, and adipose tissue.
H-MSCs are characterized by the surface markers CD105.
CD90, and CD73, while lacking hematopoietic markers such as CD34 and CD45.
When cultured under hypoxic conditions.
H-MSCs secrete a bioactive secretome enriched with growth factors and anti-inflammatory cytokines, such as interleukin-10 (IL-.
and transforming growth factor beta (TGF-), which enhance their .
These capabilities may contribute to attenuate collagen loss, by shifting the inflammatory response toward a reparative phase.
The proinflammatory mediators NF-B subunit p65 and IL-1 are pivotal in triggering the pathological cascade that impairs ECM integrity and disrupts collagen synthesis.
Recent studies have highlighted that H-MSCs, through their capacity to secrete anti-inflammatory molecules, may mitigate the harmful effects of collagen loss, through modulating proinflammatory conditions, thereby restoring ECM homeostasis.
Notably.
H-MSCs regulate the expression of -smooth muscle actin (-SMA) in myofibroblasts, a marker of tissue repair, by dynamically modulating wound healing phases.
While increased -SMA expression facilitates wound closure, prolonged expression may result in abnormal collagen synthesis and scar formation, highlighting the need for precise regulation.
Similarly.
H-MSCs play a critical role in suppressing proinflammatory conditions, ensuring that the healing process is pathological conditions.
The study by Mayasari et al.
found that treatment with the secretome from hypoxic mesenchymal stem cells (H-MSC.
resulted in decreased expression of genes IL-6 .
59 A 3.
32 pg/mL), p50 .
35 A 2.
, and p65 .
09 A 1.
82 a.
) in a hyperpigmentation animal model, compared to the control group.
The decrease in gene expression was observed to correlate with increasing concentration of the HMSC secretome.
While the previous study has shown the effects of the H-MSC secretome on IL6, p50, and p65 gene expression in a hyperpigmentation model, studies investigating its role in collagen loss models have not yet been The present study evaluated the effect of HMSC-mediated suppression of inflammatory mediators in a UVB-induced collagen degradation model, with specific focus on quantitatively assessing the dose-dependent regulatory effects of H-MSCs on NF-B p65 and IL-1 expression in Universa Medicina.
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H-MSCs Modulate Inflammation in Collagen Loss
METHODS
Research design This post-test-only experimental study was carried out at the Stem Cell and Cancer Research (SCCR) Laboratory, located in Semarang.
Central Java.
Indonesia, between October and November Experimental animals The study utilized Wistar rats (Rattus norvegicu.
weighing 200Ae250 g as an animal model for collagen loss.
They were acquired from the Animal Research Centre laboratory.
SCCR Indonesia.
Environmental conditions were carefully regulated.
The indoor temperature was maintained at 26AC, with 12-hour light/dark cycles, and humidity of 50-60%.
The animals had free access to food and water.
The feed and bedding were regularly replaced to ensure healthy and stable growth.
In this study, the sample size was determined based on FedererAos formula: .
- .
- .
Ou 15, where t is the number of treatment groups and n is the number of replications per With five groups .
and five rats per group .
, this study needed 25 male Wistar rats, each weighing between 200 and 250 g.
The rats were randomly divided into five groups based on previous research.
Preparation, phenotyping, and differentiation analysis of UC-MSCs Umbilical cord-derived mesenchymal stem cells (UC-MSC.
were isolated following established protocols.
The cells were cultured in DulbeccoAos Modified Eagle Medium (DMEM.
Gibco.
NY.
USA) supplemented with 10% fetal (FBS.
Gibc.
, penicillin/streptomycin (Gibc.
, and 0.
amphotericin B (Gibc.
at 37AC under 5% COCC The culture medium was replaced every three days.
UC-MSCs at passage 5 were used for all subsequent experiments.
Flow cytometry analysis was conducted to characterize surface markers on UC-MSCs at passage 5.
Cells were stained with rat anti-CD90-FITC.
CD29-PE.
CD31-perCP, and CD45-APC antibodies (BD Bioscience.
CA.
USA).
Analysis was performed using a BD Accuri C6 Plus flow cytometer and accompanying software (BD Bioscienc.
Adipogenic and osteogenic differentiation potential of UC-MSCs at passage 5 was assessed using standard differentiation media at 37AC with COCC.
Adipogenic differentiation basal media (MesenCultE.
Stem Cell Technologies.
Singapor.
were supplemented with specific differentiation supplements (Stem Cell Technologie.
, 1% L-glutamine (Gibc.
, 1% penicillin (Gibc.
, and 0.
25% amphotericin B (Gibc.
Media changes were performed every three days.
After 21 days of differentiation, lipid deposits were visualized using Oil Red O staining, while calcium deposits were identified using Alizarin Red staining (Sigma-Aldrich.
MO.
USA).
Induction of hypoxic environment for UCMSCs To simulate hypoxic conditions.
UC-MSCs at approximately 80% confluence were transferred to a hypoxia chamber (Stem Cell Technologie.
The chamber's oxygen concentration was maintained at 5% using an oxygen controller (BioSpherix.
Lacona.
NY.
USA), which continuously monitored the partial oxygen pressure .
OCC).
The cells were incubated under these conditions for 18 hours at 37AC with 5% COCC.
After the incubation period, the culture medium was collected for subsequent analyses.
Generation of collagen loss animal model A total of 25 healthy male Wistar rats, each weighing between 200 and 250 g, were provided with unrestricted access to food and water.
The rats were maintained under controlled conditions at a temperature of 26 AC with a 12-hour light/dark After one week of acclimatization, the rats were randomly assigned to five groups: K1 .
ealthy rat.
K2 .
ollagen los.
K3 .
ollagen loss hyaluronic acid (HA)).
K4 .
ollagen loss 5 y 10AA H-MSC.
, and K5 .
ollagen loss 5 y 10AA H-MSC.
, with five rats per group.
UVB
radiation with a peak emission wavelength of 302 nm (CL100M.
UVP.
USA) was employed to induce collagen loss.
Following a previously established protocol, the rats were exposed to 160 mJ/cmA of UVB light for 30 minutes daily over five consecutive days.
H-MSC administration All animal procedures were conducted at the Animal Research Centre.
SCCR Indonesia, in accordance with established ethical guidelines.
HMSCs were administered subcutaneously into rat skin in groups K4 and K5 by a veterinarian, using a single injection of 200 AAL NaCl.
Rats in group K4 received a dose based on previous research .
5 y 10AA H-MSCs, while those in group K5 were treated with 5 y 10AA H-MSCs.
Additionally.
Fadrian F.
Decroli E.
Ahmad A, et al rats in group K3 were administered 200 AAL of HA, while groups K1 and K2 were treated with 200 AAL NaCl.
The injections were administered under sterile conditions to ensure precision and safety.
Following administration, the rats were maintained for 14 days before being sacrificed for further analysis, to allow the H-MSCs to exert their therapeutic effects.
Sample collection On day 15, the rats were anesthetized via intramuscular injection using a combination of ketamine .
mg/kgBW) and xylazine .
mg/kgBW) to ensure proper sedation.
Following anesthesia, dorsal skin tissue samples were carefully harvested.
The collected tissue samples were preserved in RNAlaterA solution (SigmaAldric.
for subsequent qRT-PCR analysis to ensure RNA integrity.
Additional samples were fixed in 10% buffered formalin for histological Remaining tissue samples were snap-frozen and stored at -80AC for intracellular enzyme-linked immunosorbent assay (ELISA) to quantify protein expression levels.
Collagen staining Paraffin-embedded skin tissue blocks were sectioned into 5 m thick slices using a precision The sections were stained with MassonAos Trichrome stain (Bio-Optica.
Milano.
Ital.
following the manufacturerAos protocol to visualize collagen fibers.
Stained slides were examined under a light microscope (Leica.
German.
at appropriate magnifications.
The collagen density was quantified using ImageJ software, calculated as the percentage of collagenous tissue area relative to the total tissue area on each slide.
This quantitative analysis ensured an accurate assessment of collagen mRNA expression analysis Total RNA was extracted from 50 mg of dorsal skin tissue collected on day 14 using TRI Reagent (Sigma-Aldric.
, manufacturerAos protocol.
Complementary DNA .
DNA) was synthesized using the Enhanced Avian First Strand cDNA Synthesis Kit (SigmaAldric.
, adhering to the manufacturer's The reverse transcription reaction utilized oligo .
T) primers, with an incubation step at 70AC for 10 minutes, followed by 45AC for 15 minutes.
Quantitative real-time PCR .
RT- PCR) was performed in a two-step protocol using the Eco Real-Time PCR System (Illumina Inc.
San Diego.
CA.
USA) and the KAPA SYBRA FAST Universal Kit (Sigma-Aldric.
A cDNA template of 3 ng was used for each reaction.
The expression levels of p65 were assessed, with actin serving as the reference gene.
The primer sequences were as follows: -actin: forward 5'GCCTTCCTTCCTgTATG-3' and reverse 5'AGGAGCCAgCAGTAATC-3'.
p65: forward 5Ao- AACACTGCCGAGCTCAAGAT-3Ao and reverse 5Ao-CATCGGCTTGAGaGGAG-3Ao.
The thermocycling conditions included an initial denaturation step at 95AC for 3 minutes, followed by 40 cycles at 95AC for 10 seconds and at 60AC for 30 seconds.
Gene expression levels were quantified using the iiCt method, analyzed through the EcoStudy Software (Illumin.
, and normalized to the -actin housekeeping gene.
Intracellular ELISA The intracellular levels of IL-1 were measured using a specific ELISA kit, catalog number A (Elabscience.
Texas.
USA) according to the manufacturerAos protocol.
Tissue lysates were prepared from the extracted samples using a radioimmunoprecipitation (RIPA) lysis buffer containing PMSF Protease Inhibitor (Cat No.
EL SR.
, 0.
25% Trypsin Solution, sodium fluoride, and a protease inhibitor cocktail.
The lysates were centrifuged at 4AC for 10 minutes, and the supernatants were collected for analysis.
Protein concentrations were determined using UV-Vis spectrophotometry at 260 nm and 280 nm to ensure sample quality and consistency.
Equal volumes of protein from each sample were loaded into ELISA wells.
Standard curves were generated for each assay to calculate IL-1 concentrations.
Colorimetric absorbance was recorded at a wavelength of 450 nm using a microplate reader.
All measurements were conducted in triplicate to ensure the reliability and reproducibility of the Ethical clearance All procedures involving animals adhered to the ethical standards and guidelines established by the Institutional Research Bioethics of Universitas Islam Sultan Agung, under approval number 421/X/2024/Komisi Bioetik.
Statistical analysis All statistical analyses were conducted using SPSS version 26 (IBM.
New York.
USA).
For data Universa Medicina.
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H-MSCs Modulate Inflammation in Collagen Loss following a normal distribution, one-way analysis of variance (ANOVA) was performed, followed by TukeyAos post-hoc test to assess pairwise group For non-normally distributed data, the Kruskal-Wallis test was employed, followed by multiple comparison test Mann-Whitney U tests for pairwise comparisons.
Results were expressed as mean A standard deviation (SD).
A p-value of less than 0.
05 was considered statistically RESULTS H-MSC characteristics The characteristics of H-MSCs at the fifth passage were assessed based on cell morphology, membrane marker expression, and differentiation potential, following the International Society for Cellular Therapy (ISCT) Morphologically, the cells exhibited a fibroblastlike, spindle-shaped appearance (Figure 1.
Adipogenic (Figure 1.
and osteogenic (Figure 1.
differentiation assays confirmed their multilineage differentiation capacity.
After 21 days of incubation, the cells successfully differentiated into adipocytes and osteocytes, as evidenced by lipid droplet accumulation and calcium deposition, respectively, visualized with red staining.
Immunophenotyping revealed that the H-MSCs were positive for CD90 and CD29, characteristic markers of mesenchymal stem cells, while hematopoietic and endothelial markers CD45 and CD31 were not expressed, consistent with the H-MSCsAo mesenchymal origin (Figure UV-B exposure induces collagen loss in rats In this study, we successfully established a UVB-induced collagen degradation model, as validated through both macroscopic and Macroscopic observations revealed visible skin discoloration, with brown pigmentation indicative of UVBinduced damage.
Histological analysis using MassonAos Trichrome staining further confirmed collagen loss in the dermal tissue, as evidenced by the reduced blue color in UVB-exposed samples (Figure .
These findings demonstrate that UVB synthesis, leading to significant collagen This validated animal model provides a robust platform for studying UVB-induced skin damage and potential therapeutic interventions.
H-MSC injection suppresses IL-1 levels in UVB-induced collagen loss in rat skin Intracellular ELISA was performed to evaluate IL-1 levels in the skin tissue of the experimental groups.
A significant elevation of IL-1 was observed in group K2 .
94A205.
pg/mL) compared to group K1 .
99A51.
pg/mL) .
<0.
inflammatory activity resulting from UVB The administration of H-MSCs in groups K4 .
14A63.
76 pg/mL) and K5 .
80A137.
81 pg/mL) significantly reduced IL1 levels compared to group K2 .
94A205.
pg/mL) .
<0.
, demonstrating the antiinflammatory effects of H-MSC treatment.
Notably, group K5 .
80A137.
81 pg/mL), which received the higher dose of H-MSCs .
y 10AA cell.
, exhibited the most pronounced suppression of IL-1 levels among the treated groups .
<0.
On the other hand, group K3 .
52A176.
38 pg/mL) showed no significant .
>0.
difference in IL-1 levels compared to group K2 .
94A205.
80 pg/mL), suggesting that HA alone does not mitigate UVB-induced inflammation, as illustrated in Figure 3.
These findings underscore the potential of H-MSC therapy to counteract UVB-induced inflammation by reducing IL-1 expression in skin tissue.
H-MSC administration inhibits NF-B p65 transcription factor in rat skin with UVBinduced collagen loss The expression of the NF-B p65 transcription factor in skin tissue was analyzed using qRT-PCR.
Results demonstrated a significant upregulation of NF-B p65 expression in group K2 .
46A0.
50 a.
) compared to group K1 .
5A0.
36 a.
) .
<0.
In contrast, the administration of H-MSCs in groups K4 .
13A0.
50 a.
) and K5 .
72A0.
22 a.
effectively reduced NF-B p65 expression levels compared to group K2 .
46A0.
50 a.
) .
<0.
The results also showed a reduction in p65 NF-B expression in group K3 .
08A0.
48 a.
suggesting that HA also mitigates NF-B p65.
However, group K5 .
72A0.
22 a.
), which received 5 y 10AA H-MSCs, exhibited the lowest .
<0.
levels of NF-B p65 expression among all groups, as shown in Figure 4.
These findings suggest that H-MSC treatment mitigates UVBinduced inflammatory signaling by suppressing the activation of the NF-B p65 pathway, thereby offering a potential therapeutic strategy for preventing collagen loss.
Fadrian F.
Decroli E.
Ahmad A, et al
CD90 FITC
CD29 A C
CD45 E
CD31 erC Cy5.
Figure 1.
The characteristics of H-MSCs H-MSCs at passage 5 exhibited spindle-shaped, fibroblast-like morphology (A).
adipogenic (B) and osteogenic (C) differentiation was confirmed by oil red O and alizarin red staining .
ed coloratio.
Images were captured at 200y magnification.
Immunophenotyping analysis showed high expression of CD90 .
8%) and CD29 .
and low expression of CD45 .
1%) and CD31 .
1%), validating their mesenchymal origin Figure 2.
Validation of collagen loss Macroscopic and histological validation of collagen degradation induced by UVB irradiation.
Healthy rats showed smooth skin with no visible wrinkles (A), whereas UVB-irradiated rats without treatment exhibited prominent wrinkling (B).
Collagen staining using MassonAos Trichrome demonstrated higher collagen density .
ndicated by blue stainin.
in healthy rats (C) compared to a significant reduction in collagen content in UVBirradiated rats without treatment (D).
These findings confirm successful induction of collagen loss in the UVBirradiated animal model.
Universa Medicina.
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H-MSCs Modulate Inflammation in Collagen Loss IL 1 concentration .
g/mL) p65 NF kB mRNA relative expression p65 NF-kB mRNA relative expression .
Figure 3.
IL-1 level in skin tissue after 14 days intervention by treatment groups IL-1 expression was significantly increased in the collagen loss group (K2: 931.
94A205.
80 pg/mL) compared to the healthy control group (K1: 203.
99A51.
76 pg/mL) .
<0.
Treatment with HA in group K3 .
52A176.
pg/mL) did not result in a notable change in IL-1 expression compared to the collagen loss group (K.
However, the groups treated with H-MSCs (K4: 633.
14A63.
76 pg/mL and K5: 520.
80 A 137.
81 pg/mL) showed significantly lower IL-1 levels compared to the collagen loss group (K2: 931.
94A205.
80 pg/mL) .
<0.
, with the lowest IL-1 expression observed in the group receiving the higher H-MSC dose (K5: 520.
80A137.
pg/mL).
Data are presented as mean A SD.
*p<0.
K1: healthy rat group.
K2: collagen loss group.
K3:
collagen loss HA treatment group.
K4: collagen loss 2.
5 y 10AA H-MSC treatment group.
K5: collagen loss 5 y 10AA H-MSC treatment group.
Figure 4.
Gene expression of NF-B p65 in skin tissue after 14 days intervention by treatment groups NF-B expression was significantly increased in the collagen loss group (K2: 2.
46A0.
50 a.
) compared to the healthy control group (K1: 1.
50A0.
36 a.
) .
<0.
The groups treated with H-MSCs (K4: 1.
13A0.
50 a.
) and K5: 0.
72A0.
22 a.
) showed significantly lower NF-B levels compared to the collagen loss group (K2:
46A0.
50 a.
) .
<0.
, with the lowest NF-B expression observed in the group receiving the higher dose of H-MSCs (K5: 0.
72A0.
22 a.
Data are presented as mean A SD.
*p<0.
K1: healthy rats group.
K2: collagen loss group.
K3: collagen loss HA treatment group.
K4: collagen loss 2.
5 y 10AA H-MSC treatment group.
K5:
collagen loss 5 y 10AA H-MSC treatment group.
u : arbitrary units Fadrian F.
Decroli E.
Ahmad A, et al
DISCUSSION
Our findings demonstrated a significant reduction in NF-B p65 expression and IL-1 levels across all treatment groups compared to the negative controls.
Notably, the optimal reduction occurred in the group treated with a 5x10AA H-MSC dose in 200 AAL NaCl.
This aligns with a previous study showing that H-MSCs can decrease p65 expression in the hyperpigmented animal .
Another study also reported that WhartonAos jelly-derived MSCs decreased IL-1 gene expression in an AlzheimerAos rat model.
These findings suggest that H-MSCs also possess the capacity to downregulate p65 and IL-1 expression in UVB-induced animal models.
UVB radiation is a major cause of collagen inflammatory responses.
The inflammatory response induced by UVB radiation is primarily driven by the activation of the NF-B signaling pathway, particularly the p65 subunit in the epidermis, resulting in the secretion of various pro-inflammatory cytokines, such as IL-1.
previous study highlighted the critical role of NFB p65 in maintaining and expanding the inflammatory process in the skin, suggesting that modulation of this signaling pathway could UVB-induced .
The anti-inflammatory effect observed in this study could be attributed to the IL-10 secreted by the H-MSCs, which plays a crucial role in suppressing the inflammatory process associated with collagen loss, thereby downregulating NF-B p65 expression and IL-1 production.
This is in line with a previous study, which demonstrated that IL-10 suppresses inflammation via activation of intracellular proteins, such as suppressor of cytokine signaling 3 (SOCS.
IL-10 binds to its receptor, initiating the signal transducer and activator of transcription 3 (STAT-.
STAT3 then translocates to the nucleus and induces the mRNA sequence for SOCS3, which subsequently inhibits several pro-inflammatory signaling pathways, including NF-B p65.
This attenuation of NF-B p65 results in a decrease in the secretion of pro-inflammatory cytokines, including IL-1.
H-MSCs have demonstrated considerable potential in treating various forms of inflammation and promoting tissue regeneration.
Hypoxic preconditioning .
ulturing MSCs under low oxygen condition.
has emerged as a promising Studies have shown that H-MSCs enhance the secretion of anti-inflammatory cytokines and growth factors, helping regulate inflammatory processes and accelerating wound .
H-MSCs are known to increase IL-10 production, when administered at low doses in cases of ischemia/reperfusion-induced lung .
IL-10 which plays an essential role in suppressing excessive immune responses and reducing pro-inflammatory cytokine production, including IL-1 and TNF-.
By decreasing the levels of pro-inflammatory cytokines.
H-MSCs help mitigate inflammation within tissues.
Furthermore.
H-MSCs have been shown to modulate the NF-B pathway, a major pathway in the inflammatory response.
Activation of NFB p65 is associated with elevated proinflammatory cytokine expression.
By modulating this pathway.
H-MSCs can reduce NF-B activation, thereby decreasing IL-1 and other pro-inflammatory cytokines.
Beyond their antiinflammatory role.
H-MSCs also promote collagen synthesis through the secretion of growth factors such as TGF-.
Our study underscores the clinical relevance of H-MSC-based therapies as novel and potentially safer alternatives for managing UVBinduced skin damage and inflammation-related collagen loss.
By targeting key pro-inflammatory mediators and restoring extracellular matrix homeostasis.
H-MSCs offer a regenerative strategy with broad therapeutic potential in dermatology and tissue repair.
Notably, while hyaluronic acid (HA) treatment alone did not attenuate inflammatory markers.
H-MSC treatment significantly downregulated both p65 and IL-1, indicating a superior anti-inflammatory profile and highlighting the therapeutic advantage of cell-based interventions over conventional topical agents.
While this study demonstrates the promising anti-inflammatory effects of HMSCs in the rat collagen loss model, several limitations warrant consideration for future Although the study provides valuable insights into the anti-inflammatory effects of H-MSCs, the underlying molecular mechanisms, particularly how H-MSCs modulate other inflammatory pathways beyond NF-B, remain unclear.
Further research is needed to elucidate the full spectrum of cytokine interactions and cellular pathways involved in the H-MSCs anti-inflammatory and regenerative effects.
Universa Medicina.
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H-MSCs Modulate Inflammation in Collagen Loss
CONCLUSION
Our findings provide compelling evidence that subcutaneous injection of H-MSCs effectively mitigates inflammation in the rat collagen loss model by downregulating the expression of NF-B p65 and reducing IL-1 Our data underscore the therapeutic potential of H-MSCs as a targeted intervention for inflammatory disorders, particularly those associated with collagen depletion and impaired synthesis in the skin.
The anti-inflammatory and regenerative capabilities of H-MSCs, especially in modulating key inflammatory pathways and promoting tissue repair, suggest that they could offer significant clinical benefits for conditions such as photoaging.
Given the promising outcomes observed, further clinical investigations are necessary to explore the broader application of H-MSCs in the management of skin aging and other inflammatory skin diseases.
ACKNOWLEDGMENTS
Data Availability Statement Derived data supporting the findings of this study are available from the corresponding author on request.
Declaration the Use of AI in Scientific Writing This manuscript has benefited from the use of artificial intelligence (AI)-assisted tools to support the scientific writing process.
OpenAIAos ChatG T was employed for language refinement, grammar correction, summarization, paraphrasing, and formatting of scientific content.
All scientific interpretations, and conclusions presented in this work were conceived, conducted, and validated solely by the authors.
The AI was not involved in the generation of original data, analysis, or critical scientific decision-making.
The authors take full responsibility for the integrity and accuracy of the content presented.
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