Exosomal miR‑194 from adipose‑derived stem cells impedes hypertrophic scar formation through targeting TGF‑&beta;1

Xu,Zhishan; Tian,Yuan; Hao,Lijun

doi:10.3892/mmr.2024.13340

Exosomal miR‑194 from adipose‑derived stem cells impedes hypertrophic scar formation through targeting TGF‑β1

Authors:
- Zhishan Xu
- Yuan Tian
- Lijun Hao
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Affiliations: The Plastic and Cosmetic Center, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150000, P.R. China
Published online on: September 26, 2024 https://doi.org/10.3892/mmr.2024.13340
Article Number: 216
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hypertrophic scars, which result from aberrant fibrosis and disorganized collagen synthesis by skin fibroblasts, emerge due to disrupted wound healing processes. These scars present significant psychosocial and functional challenges to affected individuals. The current treatment limitations largely arise from an incomplete understanding of the underlying mechanisms of hypertrophic scar development. Recent studies, however, have shed light on the potential of exosomal non‑coding RNAs interventions to mitigate hypertrophic scar proliferation. The present study assessed the impact of exosomes derived from adipose‑derived stem cells (ADSCs‑Exos) on hypertrophic scar formation using a rabbit ear model. It employed hematoxylin and eosin staining, Masson's trichrome staining and immunohistochemical staining techniques to track scar progression. The comprehensive analysis of the present study encompassed the differential expression of non‑coding RNAs, enrichment analyses of functional pathways, protein‑protein interaction studies and micro (mi)RNA‑mRNA interaction investigations. The results revealed a marked alteration in the expression levels of long non‑coding RNAs and miRNAs following ADSCs‑Exos treatment, with little changes observed in circular RNAs. Notably, miRNA (miR)‑194 emerged as a critical regulator within the signaling pathways that govern hypertrophic scar formation. Dual‑luciferase assays indicated a significant reduction in the promoter activity of TGF‑β1 following miR‑194 overexpression. Reverse transcription‑quantitative PCR and immunoblotting assays further validated the decrease in TGF‑β1 expression in the treated samples. In addition, the treatment resulted in diminished levels of inflammatory markers IL‑1β, TNF‑α and IL‑10. In vivo evidence strongly supported the role of miR‑194 in attenuating hypertrophic scar formation through the suppression of TGF‑β1. The present study endorsed the strategic use of ADSCs‑Exos, particularly through miR‑194 modulation, as an effective strategy for reducing scar formation and lowering pro‑inflammatory and fibrotic indicators such as TGF‑β1. Therefore, the present study advocated the targeted application of ADSCs‑Exos, with an emphasis on miR‑194 modulation, as a promising approach to managing proliferative scarring.

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1	Barone N, Safran T, Vorstenbosch J, Davison PG, Cugno S and Murphy AM: Current advances in hypertrophic scar and keloid management. Semin Plast Surg. 35:145–152. 2021. View Article : Google Scholar : PubMed/NCBI
2	Finnerty CC, Jeschke MG, Branski LK, Barret JP, Dziewulski P and Herndon DN: Hypertrophic scarring: The greatest unmet challenge after burn injury. Lancet. 388:1427–1436. 2016. View Article : Google Scholar : PubMed/NCBI
3	Edwards J: Hypertrophic scar management. Br J Nurs. 31:S24–S31. 2022. View Article : Google Scholar : PubMed/NCBI
4	Frech FS, Hernandez L, Urbonas R, Zaken GA, Dreyfuss I and Nouri K: Hypertrophic scars and keloids: Advances in treatment and review of established therapies. Am J Clin Dermatol. 24:225–245. 2023. View Article : Google Scholar : PubMed/NCBI
5	Gauglitz GG, Korting HC, Pavicic T, Ruzicka T and Jeschke MG: Hypertrophic scarring and keloids: Pathomechanisms and current and emerging treatment strategies. Mol Med. 17:113–125. 2011. View Article : Google Scholar : PubMed/NCBI
6	Schmieder SJ and Ferrer-Bruker SJ: Hypertrophic scarring. StatPearls [Internet]. StatPearls Publishing; Treasure Island, FL: 2024
7	Su L and Han J: Non-coding RNAs in hypertrophic scars and keloids: Current research and clinical relevance: A review. Int J Biol Macromol. 256:1283342024. View Article : Google Scholar : PubMed/NCBI
8	Jia Q, Zhao H, Wang Y, Cen Y and Zhang Z: Mechanisms and applications of adipose-derived stem cell-extracellular vesicles in the inflammation of wound healing. Front Immunol. 1:1214757 4. 2023.
9	Romano IR, D'Angeli F, Vicario N, Russo C, Genovese C, Lo Furno D, Mannino G, Tamburino S, Parenti R and Giuffrida R: Adipose-derived mesenchymal stromal cells: A tool for bone and cartilage repair. Biomedicines. 11:17812023. View Article : Google Scholar : PubMed/NCBI
10	Lopez-Yus M, García-Sobreviela MP, Del Moral-Bergos R and Arbones-Mainar JM: Gene therapy based on mesenchymal stem cells derived from adipose tissue for the treatment of obesity and its metabolic complications. Int J Mol Sci. 24:74682023. View Article : Google Scholar : PubMed/NCBI
11	Lee JH, Won YJ, Kim H, Choi M, Lee E, Ryoou B, Lee SG and Cho BS: Adipose tissue-derived mesenchymal stem cell-derived exosomes promote wound healing and tissue regeneration. Int J Mol Sci. 24:104342023. View Article : Google Scholar : PubMed/NCBI
12	Chen Y, Younis MR, He G, Zheng Z, Wang Y, Xue K, Sun J, Liu K, Huang P and Wang X: Oxidative stimuli-responsive ‘pollen-like’ exosomes from silver nanoflowers remodeling diabetic wound microenvironment for accelerating wound healing. Adv Healthc Mater. 12:e23004562023. View Article : Google Scholar : PubMed/NCBI
13	Jing S, Li H and Xu H: Mesenchymal stem cell derived exosomes therapy in diabetic wound repair. Int J Nanomedicine. 18:2707–2720. 2023. View Article : Google Scholar : PubMed/NCBI
14	Edis A, Lumbis RH and Hedley J: Nursing management of a rabbit undergoing a rhinostomy. Vet Nurse. 7:18–24. 2016. View Article : Google Scholar
15	Feldman ER, Singh B, Mishkin NG, Lachenauer ER, Martin-Flores M and Daugherity EK: Effects of cisapride, buprenorphine, and their combination on gastrointestinal transit in New Zealand white rabbits. J Am Assoc Lab Anim Sci. 60:221–228. 2021. View Article : Google Scholar : PubMed/NCBI
16	Askar R, Fredriksson E, Manell E, Hedeland M, Bondesson U, Bate S, Olsén L and Hedenqvist P: Bioavailability of subcutaneous and intramuscular administrated buprenorphine in New Zealand white rabbits. BMC Vet Res. 16:4362020. View Article : Google Scholar : PubMed/NCBI
17	Leary S and Johnson C: AVMA guidelines for the euthanasia of animals: 2020 edition*. Members of the Panel on Euthanasia AVMA Staff Consultants; 2020
18	Suvik A and Effendy A: The use of modified Masson's trichrome staining in collagen evaluation in wound healing study. Mal J Vet Res. 3:39–47. 2012.
19	Yu G, Wang LG, Han Y and He QY: clusterProfiler: An R package for comparing biological themes among gene clusters. OMICS. 16:284–287. 2012. View Article : Google Scholar : PubMed/NCBI
20	Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, Amin N, Schwikowski B and Ideker T: Cytoscape: A software environment for integrated models of biomolecular interaction networks. Genome Res. 13:2498–2504. 2003. View Article : Google Scholar : PubMed/NCBI
21	Agarwal V, Bell GW, Nam JW and Bartel DP: Predicting effective microRNA target sites in mammalian mRNAs. eLife. 4:e050052015. View Article : Google Scholar : PubMed/NCBI
22	Chen Y and Wang X: miRDB: An online database for prediction of functional microRNA targets. Nucleic Acids Res. 48(D1): D127–D131. 2020. View Article : Google Scholar : PubMed/NCBI
23	Yang JH, Li JH, Shao P, Zhou H, Chen YQ and Qu LH: starBase: A database for exploring microRNA-mRNA interaction maps from Argonaute CLIP-Seq and Degradome-Seq data. Nucleic Acids Res. 39((Database Issue)): D202–D209. 2010.PubMed/NCBI
24	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
25	Xie S, Zhu Q, Qu W, Xu Z, Liu X, Li X, Li S, Ma W, Miao Y, Zhang L, et al: sRNAPrimerDB: Comprehensive primer design and search web service for small non-coding RNAs. Bioinformatics. 35:1566–1572. 2019. View Article : Google Scholar : PubMed/NCBI
26	Zhu Z, Hou Q, Li M and Fu X: Molecular mechanism of myofibroblast formation and strategies for clinical drugs treatments in hypertrophic scars. J Cell Physiol. 235:4109–4119. 2020. View Article : Google Scholar : PubMed/NCBI
27	Liu B, Lin L, Yu S, Xia R and Zheng L: Long non-coding RNA H19 acts as a microRNA-194 sponge to inhibit the apoptosis and promote the proliferation of hypertrophic scar fibroblasts. Can J Physiol Pharmacol. 99:1288–1297. 2021. View Article : Google Scholar : PubMed/NCBI
28	Chen SH, Chen ZY, Lin YH, Chen SH, Chou PY, Kao HK and Lin FH: Extracellular vesicles of adipose-derived stem cells promote the healing of traumatized achilles tendons. Int J Mol Sci. 22:123732021. View Article : Google Scholar : PubMed/NCBI
29	Liu Y and Holmes C: Tissue regeneration capacity of extracellular vesicles isolated from bone marrow-derived and adipose-derived mesenchymal stromal/stem cells. Front Cell Dev Biol. 9:6480982021. View Article : Google Scholar : PubMed/NCBI
30	Liu Y, Li Y, Li N, Teng W, Wang M, Zhang Y and Xiao Z: TGF-β1 promotes scar fibroblasts proliferation and transdifferentiation via up-regulating MicroRNA-21. Sci Rep. 6:322312016. View Article : Google Scholar : PubMed/NCBI
31	Liu SC, Bamodu OA, Kuo KT, Fong IH, Lin CC, Yeh CT and Chen SG: Adipose-derived stem cell induced-tissue repair or wound healing is mediated by the concomitant upregulation of miR-21 and miR-29b expression and activation of the AKT signaling pathway. Arch Biochem Biophys. 705:1088952021. View Article : Google Scholar : PubMed/NCBI
32	Zhang S and Pan S: miR-124-3p targeting of TGF-β1 inhibits the proliferation of hypertrophic scar fibroblasts. Adv Clin Exp Med. 30:263–271. 2021. View Article : Google Scholar : PubMed/NCBI
33	Yao W, Li Y, Han L, Ji X, Pan H, Liu Y, Yuan J, Yan W and Ni C: The CDR1as/miR-7/TGFBR2 axis modulates EMT in silica-induced pulmonary fibrosis. Toxicol Sci. 166:465–478. 2018. View Article : Google Scholar : PubMed/NCBI
34	Firmansyah Y, Sidharta VM, Wijaya L and Tan ST: Unraveling the significance of growth factors (TGF-β, PDGF, KGF, FGF, Pro Collagen, VEGF) in the dynamic of wound healing. Asian J Med Health. 22:49–61. 2024. View Article : Google Scholar
35	Qi J, Wu Y, Zhang H and Liu Y: LncRNA NORAD regulates scar hypertrophy via miRNA-26a mediating the regulation of TGFβR1/2. Adv Clin Exp Med. 30:395–403. 2021. View Article : Google Scholar : PubMed/NCBI
36	Yu J, Zhang L, Zhang S, Xian G, Zhao Y and Bu X: MiR-29b inhibits hypertrophic scar tissue inflammation after burn through regulating TGF-β1/Smad signaling pathway. Ital J Dermatol Venerol. 156:251–252. 2021.PubMed/NCBI
37	Qi X, Liu Y and Yang M: Circ_0057452 functions as a ceRNA in hypertrophic scar fibroblast proliferation and VEGF expression by regulating TGF-β2 expression and adsorbing miR-145-5p. Am J Transl Res. 13:6200–6210. 2021.PubMed/NCBI
38	Xu S, Dong W and Shi Y: LncRNA PICSAR binds to miR-485-5p and activates TGF-β1/Smad to promote abnormal proliferation of hypertrophic scar fibroblasts (HSFs) and excessive deposition of extracellular matrix (ECM). Med Mol Morphol. 54:337–345. 2021. View Article : Google Scholar : PubMed/NCBI
39	Pang J, Zhu L, Lv Y, Xu L, Li N, Guo P and Feng Q: miR-15a-5p up-regulates TLR4 and induces the formation of hypertrophic scars and keloids. Cell Mol Biol (Noisy-le-grand). 69:158–163. 2023. View Article : Google Scholar : PubMed/NCBI
40	Lian N and Li T: Growth factor pathways in hypertrophic scars: Molecular pathogenesis and therapeutic implications. Biomed Pharmacother. 84:42–50. 2016. View Article : Google Scholar : PubMed/NCBI
41	Zhou X, Lu J, Wu B and Guo Z: HOXA11-AS facilitates the proliferation, cell cycle process and migration of keloid fibroblasts through sponging miR-188-5p to regulate VEGFA. J Dermatol Sci. 106:111–118. 2022. View Article : Google Scholar : PubMed/NCBI
42	Chen L, Zheng Q, Liu Y, Li L, Chen X and Wang L and Wang L: Adipose-derived stem cells promote diabetic wound healing via the recruitment and differentiation of endothelial progenitor cells into endothelial cells mediated by the VEGF-PLCγ-ERK pathway. Arch Biochem Biophys. 692:1085312020. View Article : Google Scholar : PubMed/NCBI
43	Zhang YF, Li HZ, Wang XY, Ma HC, Wu Y, Yuan XH and Chu YH: Morphology of hypertrophic scar tissues and expressions of vascular endothelial growth factor and transforming growth factor beta activated kinase 1 in these tissues. Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 37:446–450. 2015.(In Chinese). PubMed/NCBI
44	Kinashi H, Ito Y, Mizuno M, Suzuki Y, Terabayashi T, Nagura F, Hattori R, Matsukawa Y, Mizuno T, Noda Y, et al: TGF-β1 promotes lymphangiogenesis during peritoneal fibrosis. J Am Soc Nephrol. 24:1627–1642. 2013. View Article : Google Scholar : PubMed/NCBI
45	Komi DEA, Khomtchouk K and Santa Maria PL: A review of the contribution of mast cells in wound healing: Involved molecular and cellular mechanisms. Clin Rev Allergy Immunol. 58:298–312. 2019. View Article : Google Scholar : PubMed/NCBI
46	Ohnuma K, Kasagi S, Uto K, Noguchi Y, Nakamachi Y, Saegusa J and Kawano S: MicroRNA-124 inhibits TNF-α- and IL-6-induced osteoclastogenesis. Rheumatol Int. 39:689–695. 2019. View Article : Google Scholar : PubMed/NCBI
47	Ma Y, Liu Z, Miao L, Jiang X, Ruan H, Xuan R and Xu S: Mechanisms underlying pathological scarring by fibroblasts during wound healing. Int Wound J. 20:2190–2206. 2023. View Article : Google Scholar : PubMed/NCBI
48	Eremenko EE, Kwan PO, Ding J, Ghosh S and Tredget EE: The effects of TGF-β1 and IFN-α2b on decorin, decorin isoforms and type I collagen in hypertrophic scar dermal fibroblasts. Wound Repair Regen. 32:135–145. 2024. View Article : Google Scholar : PubMed/NCBI