Combination nanopatterns-based Bluray discs and spidroin for directing chondrogenesis of human mesenchymal stem cells

Human Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs) have been widely used as a cell source for the development of cartilage tissue engineering. This study aims to test the effect of nanopattern-based nanotopography with a combination of Argiope appensa spider webs extract protein (Spidroin) to induce the differentiation of hWJ-MSCs into chondrocytes. Nanopattern fabrication was obtained by printed nanogrid patterns on the surface of polycarbonate Bluray-disc Recordable using Polydimethylsiloxane. The accomplishment of fabrication was confirmed through scanning electron microscopy and atomic force microscopy to determine the size of nanopatterns. Spidroin was characterized by specifying several parameters such as zeta potential, viscosity, Raman spectroscopy, water contact angle, and cytotoxicity test. The proliferation and differentiation of hWJ-MSCs cells were analyzed using 3 [4,5 - dymethylthiazol-2y1] - 2,5 - dyphenylthiazolium bromide) (MTT), Alcian Blue, and Immunocytochemistry (ICC) assays on days 7, 14, and 21. Nanopatterns were fabricated with ridge, groove, and depth pattern sizes of 145 ± 2.67, 234 ± 8.92, and 15 ± 0.782 nm, respectively. Spidroin characterization shows the presence of Arginine-Glycine-Aspartic Acid (a peptide sequence) sequences. The markers of chondrogenesis, glycosaminoglycans, and ICC for Collagen II and SOX9, were detected starting on day 7 and continued to increase until day 21. In conclusion, a combination of nanopattern and spidroin could accelerate the differentiation process of hWJ-MSCs into chondrocytes as a new method in developing cartilage tissue engineering.