Hypoxic mesenchymal stem cell‐conditioned medium accelerates wound healing by regulating IL‐10 and TGF‐β levels in a full‐thickness‐wound rat model

https://doi.org/10.22146/ijbiotech.63914

Adi Muradi Muhar(1), Faizal Mukharim(2), Dedy Hermansyah(3), Agung Putra(4*), Nurul Hidayah(5), Nur Dina Amalina(6), Iffan Alif(7)

(1) Department of Surgical, Medical Faculty, Universitas Sumatera Utara (USU), Medan 20155, Indonesia
(2) Graduate student of Medical Faculty, Sultan Agung Islamic University (UNISSULA), Semarang 50112, Indonesia
(3) Department of Surgical, Medical Faculty, Universitas Sumatera Utara (USU), Medan 20155, Indonesia
(4) Stem Cell and Cancer Research (SCCR), Medical Faculty, Sultan Agung Islamic University (UNISSULA), Semarang 50112, Indonesia; Department of Pathology, Medical Faculty, Sultan Agung Islamic University (UNISSULA), Semarang 50112, Indonesia; Department of Postgraduate Biomedical Science, Medical Faculty, Sultan Agung Islamic University (UNISSULA), Semarang 50112, Indonesia
(5) Stem Cell and Cancer Research (SCCR), Medical Faculty, Sultan Agung Islamic University (UNISSULA), Semarang 50112, Indonesia
(6) Stem Cell and Cancer Research (SCCR), Medical Faculty, Sultan Agung Islamic University (UNISSULA), Semarang 50112, Indonesia; Pharmacy Study Program, Faculty of Mathematics and Natural Sciences, Universitas Negeri Semarang, Semarang 50229, Indonesia
(7) Stem Cell and Cancer Research (SCCR), Medical Faculty, Sultan Agung Islamic University (UNISSULA), Semarang 50112, Indonesia
(*) Corresponding Author

Abstract


Full‐thickness wound healing is a complex process requiring a well‐orchestrated mechanism of various factors, including cytokines, particularly interleukin (IL)‐10 and transforming growth factor (TGF)‐β. IL‐10 and TGF‐β act as robust anti‐inflammatory cytokines in accelerating the wound healing process by regulating myofibroblasts. Hypoxic mesenchymal stem cell‐conditioned medium (hypMSC‐CM) containing cytokines potentially contribute to accelerate wound repair without scarring through the paracrine mechanism. This study aims to observe the role of hypMSC‐CM in controlling TGF‐β and IL‐10 levels to accelerate full‐thickness wound repair and regeneration. A total of 24 male Wistar rats were used in this study. Six healthy rats as a sham group and 18 rats were created as full‐thickness‐wound animal models using a 6 mm punch biopsy. The animals were randomly assigned into three groups (n = 6) consisting of two treatment groups treated with hypMSC‐CM at a low dose (200 µL hypMSC‐CM with 2 g water‐based gel added) and a high dose (400 µL hypMSC‐CM with 2 g water‐based gel added) and a control group (2 g water‐based gel only). The IL‐10 and TGF‐β levels were examined by ELISA. The results showed a significant increase in IL‐10 levels on day 3 after hypMSC‐CM treatment, followed by a decrease in platelet‐derived growth factor (PDGF) levels on days 6 and 9. In line with this finding, the TGF‐β levels also increased significantly on day 3 and then linearly decreased on days 6 and 9. HypMSC‐CM administra‐ tion may thus promote wound healing acceleration by controlling IL‐10 and TGF‐β levels in a full‐thickness‐wound rat model.


Keywords


fibroblast; hypMSC‐CM; IL‐10; TGF‐β; wound healing

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References

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improving wound repair. Int. J. Mol. Sci. 21(19):1–
15. doi:10.3390/ijms21197038.
Alagesan S, Brady J, Byrnes D, Fandiño J, Masterson C,
McCarthy S, Laffey J, O’Toole D. 2022. Enhancement strategies for mesenchymal stem cells and related therapies. Stem Cell Res. Ther. 13(1):1–13.
doi:10.1186/s13287­022­02747­w.
Batsali AK, Pontikoglou C, Koutroulakis D, Pavlaki KI,
Damianaki A, Mavroudi I, Alpantaki K, Kouvidi E,
Kontakis G, Papadaki HA. 2017. Differential expression of cell cycle and WNT pathway­related genes accounts for differences in the growth and differentiation potential of Wharton’s jelly and bone marrowderived mesenchymal stem cells. Stem Cell Res.
Ther. 8(1):1–18. doi:10.1186/s13287­017­0555­9.
Beegle J, Lakatos K, Kalomoiris S, Stewart H, Isseroff
RR, Nolta JA, Fierro FA. 2015. Hypoxic preconditioning of mesenchymal stromal cells induces
metabolic changes, enhances survival, and promotes
cell retention in vivo. Stem Cells 33(6):1818–1828.
doi:10.1002/stem.1976.
Castrén E, Sillat T, Oja S, Noro A, Laitinen A, Konttinen
YT, Lehenkari P, Hukkanen M, Korhonen M. 2015.
Osteogenic differentiation of mesenchymal stromal
cells in two­dimensional and three­dimensional cultures without animal serum. Stem Cell Res. Ther.
6(1):1–13. doi:10.1186/s13287­015­0162­6.
Darlan DM, Munir D, Putra A, Alif I, Amalina ND, Jusuf
NK, Putra IB. 2022. Revealing the decrease of indoleamine 2,3­dioxygenase as a major constituent
for B cells survival post­mesenchymal stem cells
co­cultured with peripheral blood mononuclear cell
(PBMC) of systemic lupus erythematosus (SLE) patients. Med. Glas. 19(1). doi:10.17392/1414­21.
Dominici M, Le Blanc K, Mueller I, Slaper­Cortenbach
I, Marini FC, Krause DS, Deans RJ, Keating A,
Prockop DJ, Horwitz EM. 2006. Minimal criteria for defining multipotent mesenchymal stromal
cells. The International Society for Cellular Therapy position statement. Cytotherapy. 8(4):315–317.
doi:10.1080/14653240600855905.
Drawina P, Putra A, Nasihun T, Prajoko YW, Dirja BT,
Amalina ND. 2022. Increased serial levels of plateletderived growth factor using hypoxic mesenchymal
stem cell­conditioned medium to promote closure acceleration in a full­thickness wound. Indones. J.
Biotechnol. 27(1):36. doi:10.22146/ijbiotech.64021.
El Agha E, Kramann R, Schneider RK, Li X, Seeger
W, Humphreys BD, Bellusci S. 2017. Mesenchymal Stem Cells in Fibrotic Disease. Cell Stem Cell
21(2):166–177. doi:10.1016/j.stem.2017.07.011.
Hamra NF, Putra A, Tjipta A, Amalina ND, Nasihun T.
2021. Hypoxia mesenchymal stem cells accelerate
wound closure improvement by controlling α­smooth
muscle actin expression in the full­thickness animal
model. Open Access Maced. J. Med. Sci. 9:35–41.
doi:10.3889/oamjms.2021.5537.
He X, Dong Z, Cao Y, Wang H, Liu S, Liao L, Jin Y, Yuan
L, Li B, Bolontrade MF. 2019. MSC­Derived Exosome Promotes M2 Polarization and Enhances Cutaneous Wound Healing. Stem Cells Int. 2019:1–16.
doi:10.1155/2019/7132708.
Ho CH, Lan CW, Liao CY, Hung SC, Li HY, Sung YJ.
2018. Mesenchymal stem cells and their conditioned
medium can enhance the repair of uterine defects in



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