TNF-α-Activated MSC-CM Topical Gel Effective in Increasing PDGF Level, Fibroblast Density, and Wound Healing Process Compared to Subcutaneous Injection Combination

Novalia Kuntardjo, Edi Dharmana, Chodidjah Chodidjah, Taufiq R Nasihun, Agung Putra

Abstract


Mesenchymal stem cells (MSCs) are multipotent stromal cells that have the capacity to regenerate tissue damage. However, they have several limitations. MSC-CM as a new approach treatment is widely used to solve the limitation of MSC in wound healing. The aim of this study was to evaluate the effectiveness of TNF-α-activated MSC-CM topical gel compare to topical-subcutaneous injection combination on wound healing acceleration. This study was conducted between April and August 2018 at the Stem Cell and Cancer Research Laboratory (SCCR), Faculty of Medicine, Sultan Agung Islamic University, Semarang. Experimental post-test only control group design was performed by involving 36 animal models randomly divided into six groups; T1, T2 (MSC-CM in topical gel 100 μL; 200 μL); ST1, ST2 (MSC-CM in subcutaneous injection : topical gel = 80 μL:20 μL; 160 μL:40 μL); CT (200 μL medium free TNF-α); CST (PBS in subcutaneous injection : topical gel = 160 μL :40 μL). The measurement of PDGF level on day 3 and 6 was conducted using ELISA assay while the fibroblast density was analyzed by light microcopy. It was found that there was was a significant increase in PDGF and fibroblast density on day 6 in the topical group when compared to the combination group (p<0,05). It is concluded that the MSC-CM topical gel is more effective than combination of topical-subcutaneous injection.

Key words: Combination, fibroblast, MSC-CM, PDGF, subcutaneous MSC-CM, topical MSC-CM

 

MSC-CM Topikal yang diaktivasi TNF-α Efektif Dalam Peningkatan Level PDGF, Densitas Fibroblast, dan Mempercepat Penyembuhan Luka dibanding dengan Kombinasi Injeksi Subkutan

Mesenchymal stem cells (MSCs) merupakan sel stroma multipoten yang memiliki kemampuan untuk meregenerasi kerusakan jaringan. Namun, MSC memiliki beberapa keterbatasan. MSC-CM sebagai terapi pendekatan baru digunakan untuk mengatasi keterbatasan MSC dalam penyembuhan luka. Tujuan penelitian ini adalah menilai efektivitas MSC-CM topikal yang diaktvasi TNF-α dibandingdengan kombinasi topikal-injeksi subkutan pada percepatan penyembuhan luka. Penelitian ini dilaksanakan pada bulan April–Agustus 2018 di Laboratorium Stem Cell and Cancer Research (SCCR), Fakultas Kedokteran, Universitas Islam Sultan Agung, Semarang. Penelitian menggunakan eksperimen laboratorium dengan rancangan post-test only control group, menggunakan 36 tikus galur wistar yang dibagi secara acak menjadi 6 kelompok; T1, T2 (MSC-CM gel topikal 100μL; 200μL); ST1, ST2 (MSC-CM injeksi subkutan : gel topikal = 80 μL:20 μL; 160 μL:40 μL); CT (200 μL medium tanpa TNF-α); CST (PBS injeksi subkutan: gel topikal = 160 μL :40 μL). Pengukuran kadar PDGF pada hari ke-3 dan ke-6 mengunakan ELISA, sedangkan jumlah fibroblas dilihat mengunakan mikroskop cahaya. Hasil penelitian ini menunjukkan peningkatan kadar PDGF dan jumlah fibroblas yang signifikan dihari ke-6 pada MSC-CM gel topikal dibanding dengan kombinasi topical-injeksi subukutan (p <0.05). Simpulan penelitian ini adalah pemberian MSC-CM secara topical lebih efektif dibanding dengan kombinasi topikal-injeksi subkutan.

Kata kunci: Fibroblas, konditional medium, MSC-CM kombinasi, MSC-CM topikal, PDGF


Keywords


Combination, fibroblast, MSC-CM, PDGF, subcutaneous MSC-CM, topical MSC-CM

Full Text:

PDF

References


Prockop DJ, Youn OJ. Mesenchymal stem/stromal cells (MSCs): role as guardians of inflammation. Mol Ther. 2012;20(1):14–20.

Pang C, Ibrahim A, Bulstrode NW, Ferretti P. An overview of the therapeutic potential of regenerative medicine in cutaneous wound healing: advances and limitations in regenerative medicine for stimulating wound repair. Int Wound J. 2017;14(3):450–9.

Chen X, Wang S, Cao W. Mesenchymal stem cell-mediated immunomodulation in cell therapy of neurodegenerative diseases. Cell Immunol. 2018;326:8–14.

Tsai HW, Wang PH, Tsui KH. Mesenchymal stem cell in wound healing and regeneration. J Chinese Med Assoc. 2018;81(3):223–34.

Klinker MW. Mesenchymal stem cells in the treatment of inflammatory and autoimmune diseases in experimental animal models. World J Stem Cells. 2015;7(3):556.

Putra A, Ridwan F, Putridewi A, Kustiyah A, Wirastuti K, Sadyah NA, et al. The role of TNF-α induced MSCs on suppressive inflammation by increasing TGF-β and IL-10. Open Access Maced J Med Sci. 2018;6(10):1779–83.

Ho CH, Lan CW, Liao CY, Hung SC, Li HY, Sung YJ. Mesenchymal stem cells and their conditioned medium can enhance the repair of uterine defects in a rat model. J Chinese Med Assoc. 2018;81(3):268–76.

Meiliana A, Dewi NM, Wijaya A. Mesenchymal stem cells manage endogenous tissue regeneration. Indonesian Biomed J. 2011; 8(2):71–90.

Andrae J, Gallini R, and Betsholtz C. Role of platelet-derived growth factors in physiology and medicine. Genes Dev. 2008;22:1276–312.

Raica M, Cimpean AM. Platelet-derived growth factor (PDGF)/PDGF receptors (PDGFR) axis as target for antitumor and antiangiogenic therapy. Pharmaceuticals. 2010;3:572–99

Zhao J, Hu L, Liu J, Gong N, Chen L. The effects of cytokines in adipose stem cell-conditioned medium on the migration and proliferation of skin fibroblasts in vitro. Biomed Res Int. 2013;2013:578479.

Kwon YW, Heo SC, Jeong GO, Yoon JW, Mo WM, Lee MJ, et al. Tumor necrosis factor-α-activated mesenchymal stem cells promote endothelial progenitor cell homing and angiogenesis. Biochim Biophys Acta-Mol Basis Dis. 2013;1832(12):2136–44,

Lee MJ, Kim J, Lee K Il, Shin JM, Chae J Il, Chung HM. Enhancement of wound healing by secretory factors of endothelial precursor cells derived from human embryonic stem cells. Cytotherapy. 2011;13(2):165–78.

Chen L, Tredget EE, Wu PY, Wu Y. Paracrine factors of mesenchymal stem cells recruit macrophages and endothelial lineage cells and enhance wound healing. PLoS One. 2008;3(4):e1886.

Haque N, Abu KNH, Rahman MT. Optimization of pre-transplantation conditions to enhance the efficacy of mesenchymal stem cells. Int J Biol Sci. 2015;11(3):324–34.

Choi JW, Kim S, Kim TM, Kim YM, Seo HW, Park TS, et al. Basic fibroblast growth factor activates MEK/ERK Cell signaling pathway and stimulates the proliferation of chicken primordial germ cells. PLoS One. 2010;5(9):e12968.

Deng X, Jin K, Li Y, Gu W, Liu M, Zhou L. Platelet-Derived growth factor and transforming growth factor β1 regulate ARDS-Associated lung fibrosis through distinct signaling pathways. Cell Physiol Biochem. 2015;36(3):937–46.

Fong GH. Mechanisms of adaptive angiogenesis to tissue hypoxia. Angiogenesis. 2008;11(2):121–40.

Chen J, Li Y, Hao H, Li C, Du Y, Hu Y, et al. Mesenchymal stem cell conditioned medium promotes proliferation and migration of alveolar epithelial cells under septic conditions in vitro via the JNK-P38 signaling pathway. Cell Physiol Biochem. 2015;37(5):1830–46.




DOI: https://doi.org/10.15395/mkb.v51n1.1479

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.



 

This Journal indexed by

           


Creative Commons License
MKB is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License

 


View My Stats