In vitro study of cartilage tissue engineering using human adipose-derived stem cells induced by platelet-rich plasma and cultured on silk fibroin scaffold
Rosadi I.a,b, Karina K.b,c, Rosliana I.b, Sobariah S.b, Afini I.b, Widyastuti T.b, Barlian A.a
a School of Life Sciences and Technology, Institut Teknologi Bandung, Bandung, West Java, Indonesia
b HayandraLab, Yayasan Hayandra Peduli, Jakarta, DKI Jakarta, Indonesia
c Biomedic, Universitas Indonesia, Jakarta, DKI Jakarta, Indonesia
Abstract
© 2019 The Author(s).Background: Cartilage tissue engineering is a promising technique for repairing cartilage defect. Due to the limitation of cell number and proliferation, mesenchymal stem cells (MSCs) have been developed as a substitute to chondrocytes as a cartilage cell-source. This study aimed to develop cartilage tissue from human adipose-derived stem cells (ADSCs) cultured on a Bombyx mori silk fibroin scaffold and supplemented with 10% platelet-rich plasma (PRP). Methods: Human ADSCs and PRP were characterized. A silk fibroin scaffold with 500 μm pore size was fabricated through salt leaching. ADSCs were then cultured on the scaffold (ADSC-SS) and supplemented with 10% PRP for 21 days to examine cell proliferation, chondrogenesis, osteogenesis, and surface marker expression. The messenger ribonucleic acid (mRNA) expression of type 2 collagen, aggrecan, and type 1 collagen was analysed. The presence of type 2 collagen confirming chondrogenesis was validated using immunocytochemistry. The negative and positive controls were ADSC-SS supplemented with 10% foetal bovine serum (FBS) and ADSC-SS supplemented with commercial chondrogenesis medium, respectively. Results: Cells isolated from adipose tissue were characterized as ADSCs. Proliferation of the ADSC-SS PRP was significantly increased (p < 0.05) compared to that of controls. Chondrogenesis was observed in ADSC-SS PRP and was confirmed through the increase in glycosaminoglycans (GAG) and transforming growth factor-β1 (TGF-β1) secretion, the absence of mineral deposition, and increased surface marker proteins on chondrogenic progenitors. The mRNA expression of type 2 collagen in ADSC-SS PRP was significantly increased (p < 0.05) compared to that in the negative control on days 7 and 21; however, aggrecan was significantly increased on day 14 compared to the controls. ADSC-SS PRP showed stable mRNA expression of type 1 collagen up to 14 days and it was significantly decreased on day 21. Confocal analysis showed the presence of type 2 collagen in the ADSC-SS PRP and positive control groups, with high distribution outside the cells forming the extracellular matrix (ECM) on day 21. Conclusion: Our study showed that ADSC-SS with supplemented 10% PRP medium can effectively support chondrogenesis of ADSCs in vitro and promising for further development as an alternative for cartilage tissue engineering in vivo.
"Author
Adipose Tissue,Aggrecans,Cartilage,Cell Differentiation,Cell Proliferation,Chondrogenesis,Collagen Type I,Collagen Type II,Extracellular Matrix,Fibroins,Glycosaminoglycans,Humans,Mesenchymal Stem Cells,Osteogenesis,Platelet-Rich Plasma,Tissue Engineering,Tissue Scaffolds,Transforming Growth Factor beta1
"Indexed
Aggrecan,Chondrogenesis of ADSCs,Salt-leached scaffold,Type 1 collagen,Type 2 collagen