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

SUMMARY

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-a-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-a); 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-a Efektif Dalam Peningkatan Level PDGF, Densitas Fibroblast, dan Mempercepat Penyembuhan Luka dibanding dengan Kombinasi Injeksi SubkutanMesenchymal 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-a 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-a); 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