Inhibition and reversal of a TGF-β1 induced myofibroblast phenotype by adipose tissue-derived paracrine factors.
Myofibroblasts
/ metabolism
Transforming Growth Factor beta1
/ pharmacology
Adipose Tissue
/ cytology
Cell Differentiation
/ drug effects
Culture Media, Conditioned
/ pharmacology
Humans
Hepatocyte Growth Factor
/ pharmacology
Paracrine Communication
/ drug effects
Phenotype
Cells, Cultured
Fibroblasts
/ metabolism
Adipocytes
/ metabolism
Stromal Cells
/ metabolism
Adipose-derived stromal cells
Autologous fat grafting
Myofibroblasts
Scarring
Transforming growth factor β-1
Journal
Stem cell research & therapy
ISSN: 1757-6512
Titre abrégé: Stem Cell Res Ther
Pays: England
ID NLM: 101527581
Informations de publication
Date de publication:
13 Jun 2024
13 Jun 2024
Historique:
received:
29
03
2024
accepted:
27
05
2024
medline:
13
6
2024
pubmed:
13
6
2024
entrez:
12
6
2024
Statut:
epublish
Résumé
Hypertrophic scarring results from myofibroblast differentiation and persistence during wound healing. Currently no effective treatment for hypertrophic scarring exists however, autologous fat grafting has been shown to improve scar elasticity, appearance, and function. The aim of this study was to understand how paracrine factors from adipose tissues and adipose-derived stromal cells (ADSC) affect fibroblast to myofibroblast differentiation. The transforming growth factor-β1 (TGF-β1) induced model of myofibroblast differentiation was used to test the effect of conditioned media from adipose tissue, ADSC or lipid on the proportion of fibroblasts and myofibroblasts. Adipose tissue conditioned media inhibited the differentiation of fibroblasts to myofibroblasts but this inhibition was not observed following treatment with ADSC or lipid conditioned media. Hepatocyte growth factor (HGF) was readily detected in the conditioned medium from adipose tissue but not ADSC. Cells treated with HGF, or fortinib to block HGF, demonstrated that HGF was not responsible for the inhibition of myofibroblast differentiation. Conditioned media from adipose tissue was shown to reduce the proportion of myofibroblasts when added to fibroblasts previously treated with TGF-β1, however, conditioned media treatment was unable to significantly reduce the proportion of myofibroblasts in cell populations isolated from scar tissue. Cultured ADSC or adipocytes have been the focus of most studies, however, this work highlights the importance of considering whole adipose tissue to further our understanding of fat grafting. This study supports the use of autologous fat grafts for scar treatment and highlights the need for further investigation to determine the mechanism.
Sections du résumé
BACKGROUND
BACKGROUND
Hypertrophic scarring results from myofibroblast differentiation and persistence during wound healing. Currently no effective treatment for hypertrophic scarring exists however, autologous fat grafting has been shown to improve scar elasticity, appearance, and function. The aim of this study was to understand how paracrine factors from adipose tissues and adipose-derived stromal cells (ADSC) affect fibroblast to myofibroblast differentiation.
METHODS
METHODS
The transforming growth factor-β1 (TGF-β1) induced model of myofibroblast differentiation was used to test the effect of conditioned media from adipose tissue, ADSC or lipid on the proportion of fibroblasts and myofibroblasts.
RESULTS
RESULTS
Adipose tissue conditioned media inhibited the differentiation of fibroblasts to myofibroblasts but this inhibition was not observed following treatment with ADSC or lipid conditioned media. Hepatocyte growth factor (HGF) was readily detected in the conditioned medium from adipose tissue but not ADSC. Cells treated with HGF, or fortinib to block HGF, demonstrated that HGF was not responsible for the inhibition of myofibroblast differentiation. Conditioned media from adipose tissue was shown to reduce the proportion of myofibroblasts when added to fibroblasts previously treated with TGF-β1, however, conditioned media treatment was unable to significantly reduce the proportion of myofibroblasts in cell populations isolated from scar tissue.
CONCLUSIONS
CONCLUSIONS
Cultured ADSC or adipocytes have been the focus of most studies, however, this work highlights the importance of considering whole adipose tissue to further our understanding of fat grafting. This study supports the use of autologous fat grafts for scar treatment and highlights the need for further investigation to determine the mechanism.
Identifiants
pubmed: 38867276
doi: 10.1186/s13287-024-03776-3
pii: 10.1186/s13287-024-03776-3
doi:
Substances chimiques
Transforming Growth Factor beta1
0
Culture Media, Conditioned
0
Hepatocyte Growth Factor
67256-21-7
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
166Subventions
Organisme : EPSRC Doctoral Training Programme Scholarship
ID : EP/R513313/1
Informations de copyright
© 2024. The Author(s).
Références
Marshall CD, Hu MS, Leavitt T, Barnes LA, Lorenz HP, Longaker MT. Cutaneous scarring: basic science, current treatments, and future directions. Adv Wound Care (New Rochelle). 2018;7(2):29–45.
pubmed: 29392092
doi: 10.1089/wound.2016.0696
Van Baar ME, Essink-Bot ML, Oen IMMH, Dokter J, Boxma H, Van Beeck EF. Functional outcome after burns: a review. Burns. 2006;32(1):1–9.
pubmed: 16376020
doi: 10.1016/j.burns.2005.08.007
Hoogewerf CJ, van Baar ME, Middelkoop E, van Loey NE. Impact of facial burns: relationship between depressive symptoms, self-esteem and scar severity. Gen Hosp Psychiatry. 2014;36(3):271–6.
pubmed: 24417954
doi: 10.1016/j.genhosppsych.2013.12.001
Leventhal D, Furr M, Reiter D. Treatment of keloids and hypertrophic scars: a meta-analysis and review of the literature. Arch Facial Plast Surg. 2006;8(6):362–8.
pubmed: 17116782
doi: 10.1001/archfaci.8.6.362
Shockley WW. Scar revision techniques: Z-plasty, W-plasty, and geometric broken line closure. Facial Plast Surg Clin N Am. 2011;19(3):455–63.
doi: 10.1016/j.fsc.2011.06.002
Klinger M, Marazzi M, Vigo D, Torre M. Fat injection for cases of severe burn outcomes: a new perspective of scar remodeling and reduction. Aesthet Plast Surg. 2008;32(3):465–9.
doi: 10.1007/s00266-008-9122-1
Rigotti G, Marchi A, Galiè M, Baroni G, Benati D, Krampera M, Pasini A, Sbarbati A. Clinical treatment of radiotherapy tissue damage by lipoaspirate transplant: a healing process mediated by adipose-derived adult stem cells. Plast Reconstr Surg. 2007;119(5):1409–22.
pubmed: 17415234
doi: 10.1097/01.prs.0000256047.47909.71
Eto H, Suga H, Matsumoto D, Inoue K, Aoi N, Kato H, Araki J, Yoshimura K. Characterization of structure and cellular components of aspirated and excised adipose tissue. Plast Reconstr Surg. 2009;124(4):1087–97.
pubmed: 19935292
doi: 10.1097/PRS.0b013e3181b5a3f1
Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 2001;7(2):211–28.
pubmed: 11304456
doi: 10.1089/107632701300062859
Spiekman M, van Dongen JA, Willemsen JC, Hoppe DL, van der Lei B, Harmsen MC. The power of fat and its adipose-derived stromal cells: emerging concepts for fibrotic scar treatment. J Tissue Eng Regen Med. 2017;11(11):3220–35.
pubmed: 28156060
pmcid: 5724515
doi: 10.1002/term.2213
Tonnard P, Verpaele A, Peeters G, Hamdi M, Cornelissen M, Declercq H. Nanofat grafting: basic research and clinical applications. Plast Reconstr Surg. 2013;132(4):1017–26.
pubmed: 23783059
doi: 10.1097/PRS.0b013e31829fe1b0
van Dongen JA, Getova V, Brouwer LA, Liguori GR, Sharma PK, Stevens HP, van der Lei B, Harmsen MC. Adipose tissue-derived extracellular matrix hydrogels as a release platform for secreted paracrine factors. J Tissue Eng Regen Med. 2019;13(6):973–85.
pubmed: 30808068
pmcid: 6593768
doi: 10.1002/term.2843
Hoerst K, van den Broek L, Sachse C, Klein O, von Fritschen U, Gibbs S, Hedtrich S. Regenerative potential of adipocytes in hypertrophic scars is mediated by myofibroblast reprogramming. J Mol Med. 2019;97(6):761–75.
pubmed: 30891616
doi: 10.1007/s00109-019-01772-2
Kruger MJ, Conradie MM, Conradie M, Van De Vyver M. ADSC-conditioned media elicit an ex vivo anti-inflammatory macrophage response. J Mol Endocrinol. 2018;61(4):173–84.
pubmed: 30038054
doi: 10.1530/JME-18-0078
Mou S, Zhou M, Li Y, Wang J, Yuan Q, Xiao P, Sun J, Wang Z. Extracellular vesicles from human adipose-derived stem cells for the improvement of angiogenesis and fat-grafting application. Plast Reconstr Surg. 2019;144(4):869–80.
pubmed: 31568294
doi: 10.1097/PRS.0000000000006046
Spiekman M, Przybyt E, Plantinga JA, Gibbs S, van der Berend L, Harmsen MC. Adipose tissue–derived stromal cells inhibit TGF-β1–induced differentiation of human dermal fibroblasts and keloid scar–derived fibroblasts in a paracrine fashion. Plast Reconstr Surg. 2014;134(4):699–712.
pubmed: 25357030
doi: 10.1097/PRS.0000000000000504
Mazini L, Ezzoubi M, Malka G. Overview of current adipose-derived stem cell (ADSCs) processing involved in therapeutic advancements: flow chart and regulation updates before and after COVID-19. Stem Cell Res Ther. 2021;12(1):1–17.
pubmed: 33397467
pmcid: 7781178
doi: 10.1186/s13287-020-02006-w
Desmouliére A, Geinoz A, Gabbiani F, Gabbiani G. Transforming growth factor-β1 induces α-smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent and growing cultured fibroblasts. J Cell Biol. 1993;122(1):103–11.
pubmed: 8314838
doi: 10.1083/jcb.122.1.103
Darby IA, Zakuan N, Billet F, Desmoulière A. The myofibroblast, a key cell in normal and pathological tissue repair. Cell Mol Life Sci. 2016;73(6):1145–57.
pubmed: 26681260
doi: 10.1007/s00018-015-2110-0
Desmouléire A, Redard M, Darby I, Gabbiani G. Apoptosis mediates the decrease in cellularity during the transition between granulation tissue and scar. Am J Pathol. 1995;146(1):56–66.
Aarabi S, Bhatt KA, Shi Y, Paterno J, Chang EI, Loh SA, Holmes JW, Longaker MT, Yee H, Gurtner GC. Mechanical load initiates hypertrophic scar formation through decreased cellular apoptosis. FASEB J. 2007;21(12):3250–61.
pubmed: 17504973
doi: 10.1096/fj.07-8218com
Howard EW, Crider BJ, Updike DL, Bullen EC, Parks EE, Haaksma CJ, Sherry DM, Tomasek JJ. MMP-2 expression by fibroblasts is suppressed by the myofibroblast phenotype. Exp Cell Res. 2012;318(13):1542–53.
pubmed: 22449415
pmcid: 4164435
doi: 10.1016/j.yexcr.2012.03.007
Petrov VV, Fagard RH, Lijnen PJ. Stimulation of collagen production by transforming growth factor-β1 during differentiation of cardiac fibroblasts to myofibroblasts. Hypertension. 2002;39(2):258–63.
pubmed: 11847194
doi: 10.1161/hy0202.103268
Neves LMG, Wilgus TA, Bayat A. In vitro, ex vivo, and in vivo approaches for investigation of skin scarring: human and animal models. Adv Wound Care (New Rochelle). 2023;12(2):97–116.
pubmed: 34915768
doi: 10.1089/wound.2021.0139
Tai Y, Woods EL, Dally J, Kong D, Steadman R, Moseley R, Midgley AC. Myofibroblasts: function, formation, and scope of molecular therapies for skin fibrosis. Biomolecules. 2021;11(8):1–27.
doi: 10.3390/biom11081095
Bullock AJ, Higham MC, MacNeil S. Use of human fibroblasts in the development of a xenobiotic-free culture and delivery system for human keratinocytes. Tissue Eng. 2006;12(2):245–55.
pubmed: 16548683
doi: 10.1089/ten.2006.12.245
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2
pubmed: 11846609
doi: 10.1006/meth.2001.1262
Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A. Fiji: an open-source platform for biological-image analysis. Nat Methods. 2012;9(7):676–82.
pubmed: 22743772
doi: 10.1038/nmeth.2019
Higginbotham S. Fat, fibroblasts, and fibrosis; how deposits of adipose tissue ameliorate dermal scarring. The University of Sheffield; 2022.
Yang J, Dai C, Liu Y. Hepatocyte growth factor suppresses renal interstitial myofibroblast activation and intercepts Smad signal transduction. Am J Pathol. 2003;163(2):621–32.
pubmed: 12875981
pmcid: 1868195
doi: 10.1016/S0002-9440(10)63689-9
Sohn SH, Kim B, Sul HJ, Choi BY, Kim HS, Zang DY. Foretinib inhibits cancer stemness and gastric cancer cell proliferation by decreasing CD44 and c-MET signalling. Onco Targets Ther. 2020;13:1027–35.
pubmed: 32099405
pmcid: 7006849
doi: 10.2147/OTT.S226951
Ejaz A, Epperly MW, Hou W, Greenberger JS, Rubin JP. Adipose-derived stem cell therapy ameliorates ionizing irradiation fibrosis via hepatocyte growth factor-mediated transforming growth factor-β downregulation and recruitment of bone marrow cells. Stem Cells. 2019;37(6):791–802.
pubmed: 30861238
doi: 10.1002/stem.3000
Eder JP, Shapiro GI, Appleman LJ, Zhu AX, Miles D, Keer H, Cancilla B, Chu F, Hitchcock-Bryan S, Sherman L, McCallum S, Heath EL, Boerner SA, LoRusso PM. A phase I study of foretinib, a multi-targeted inhibitor of c-Met and vascular endothelial growth factor receptor 2. Clin Cancer Res. 2010;16(13):3507–16.
pubmed: 20472683
doi: 10.1158/1078-0432.CCR-10-0574
Liguori TTA, Liguori GR, Moreira LFP, Harmsen MC. Fibroblast growth factor-2, but not the adipose tissue-derived stromal cells secretome, inhibits TGF-β1-induced differentiation of human cardiac fibroblasts into myofibroblasts. Sci Rep. 2018;8(1):1–10.
doi: 10.1038/s41598-018-34747-3
Evans RA, Tian YC, Steadman R, Phillips AO. TGF-β1-mediated fibroblast-myofibroblast terminal differentiation—the role of Smad proteins. Exp Cell Res. 2003;282(2):90–100.
pubmed: 12531695
doi: 10.1016/S0014-4827(02)00015-0
Dally J, Khan JS, Voisey A, Charalambous C, John HL, Woods EL, Steadman R, Moseley R, Midgley AC. Hepatocyte growth factor mediates enhanced wound healing responses and resistance to transforming growth factor-β1-driven myofibroblast differentiation in oral mucosal fibroblasts. Int J Mol Sci. 2017;18(9):1–15.
doi: 10.3390/ijms18091843
Hecker L, Jagirdar R, Jin T, Thannickal VJ. Reversible differentiation of myofibroblasts by MyoD. Exp Cell Res. 2011;317(13):1914–21.
pubmed: 21440539
pmcid: 3123424
doi: 10.1016/j.yexcr.2011.03.016
Kato K, Logsdon NJ, Shin YJ, Palumbo S, Knox A, Irish JD, Roundseville SP, Rummel SR, Mohamed M, Ahmad K, Trinh JM, Kurundkar D, Knox KS, Thannickal VJ, Hecker L. Impaired myofibroblast dedifferentiation contributes to nonresolving fibrosis in aging. Am J Respir Cell Mol Biol. 2020;62(5):633–44.
pubmed: 31962055
pmcid: 7193787
doi: 10.1165/rcmb.2019-0092OC
Plikus MV, Guerrero-juarez CF, Ito M, Li YR, Priya H, Zheng Y, Shao M, Gay DL, Ramos R, Hsi TC, Oh JW, Wang X, Ramirez A, Konopelski SE, Elzein A, Wang A, Supapannachart RJ, Lee H, Lim CH, Nace A, Guo A, Treffeisen E, Andl T, Ramirez RN, Murad R, Offermanns A, Metzger D, Chambon P, Widgerow AD, Tuan T, Mortazvi A, Gupta RK, Hamilton BA, Millar SE, Seale P, Pear WS, Lazar MA, Cotsarelis G. Regeneration of fat cells from myofibroblast during wound healing. Science. 2017;355(6326):748–52.
pubmed: 28059714
pmcid: 5464786
doi: 10.1126/science.aai8792
Moulin V, Larochelle S, Langlois C, Thibault I, Lopez-Vallé CA, Roy M. Normal skin wound and hypertrophic scar myofibroblasts have differential responses to apoptotic inductors. J Cell Physiol. 2004;198(3):350–8.
pubmed: 14755540
doi: 10.1002/jcp.10415
Fayzullin A, Ignatieva N, Zakharkina O, Tokarev M, Mudryak D, Khristidis Y, Balyasin M, Kurkov A, Churbanov S, Dyuzheva T, Timashev P, Guller A, Shekhter A. Modeling of old scars: histopathological, biochemical and thermal analysis of the scar tissue maturation. Biology. 2021;10(136):1–15.
Ma Y, Barnes SP, Chen YY, Moiemen NS, Lord JM, Sardeli AV. Influence of scar age, laser type and laser treatment intervals on adult burn scars: a systematic review and meta-analysis. PLoS ONE. 2023;18:1–16.
doi: 10.1371/journal.pone.0292097
Ţuţuianu R, Roşca AM, Florea G, Prună V, Iacomi DM, Rădulescu LA, Neagu TP, Lascăr I, Titorencu ID. Heterogeneity of human fibroblasts isolated from hypertrophic scar. Rom J Morphol Embryol. 2019;60(3):793–802.
pubmed: 31912089
Nedelec B, Shankowsky H, Scott GP, Ghahary A, Tredget EE. Myofibroblasts and apoptosis in human hypertrophic scars: the effect of interferon-α2b. Surgery. 2001;130(5):798–808.
pubmed: 11685189
doi: 10.1067/msy.2001.116453