Cartilage tissue engineering, particularly micropattern, can influence the biophysical properties of mesenchymal stem cells (MSCs) leading to chondrogenesis. In this research, human Wharton’s jelly MSCs (hWJ-MSCs) were grown on a striped micropattern containing spider silk protein (spidroin) from Argiope appensa. This research aims to direct hWJ-MSCs chondrogenesis using micropattern made of spidroin bioink as opposed to fibronectin that often used as the gold standard. Cells were cultured on striped micropattern of 500 µm and 1000 µm width sizes without chondrogenic differentiation medium for 21 days. The immunocytochemistry result showed that spidroin contains RGD sequences and facilitates cell adhesion via integrin β1. Chondrogenesis was observed through the expression of glycosaminoglycan, type II collagen, and SOX9. The result on glycosaminoglycan content proved that 1000 µm was the optimal width to support chondrogenesis. Spidroin micropattern induced significantly higher expression of SOX9 mRNA on day-21 and SOX9 protein was located inside the nucleus starting from day-7. COL2A1 mRNA of spidroin micropattern groups was downregulated on day-21 and collagen type II protein was detected starting from day-14. These results showed that spidroin micropattern enhances chondrogenic markers while maintains long-term upregulation of SOX9, and therefore has the potential as a new method for cartilage tissue engineering.
Cartilage tissue engineering, particularly micropattern, can influence the biophysical properties of mesenchymal stem cells (MSCs) leading to chondrogenesis. In this research, human Wharton’s jelly MSCs (hWJ-MSCs) were grown on a striped micropattern containing spider silk protein (spidroin) from Argiope appensa. This research aims to direct hWJ-MSCs chondrogenesis using micropattern made of spidroin bioink as opposed to fibronectin that often used as the gold standard. Cells were cultured on striped micropattern of 500 µm and 1000 µm width sizes without chondrogenic differentiation medium for 21 days. The immunocytochemistry result showed that spidroin contains RGD sequences and facilitates cell adhesion via integrin β1. Chondrogenesis was observed through the expression of glycosaminoglycan, type II collagen, and SOX9. The result on glycosaminoglycan content proved that 1000 µm was the optimal width to support chondrogenesis. Spidroin micropattern induced significantly higher expression of SOX9 mRNA on day-21 and SOX9 protein was located inside the nucleus starting from day-7. COL2A1 mRNA of spidroin micropattern groups was downregulated on day-21 and collagen type II protein was detected starting from day-14. These results showed that spidroin micropattern enhances chondrogenic markers while maintains long-term upregulation of SOX9, and therefore has the potential as a new method for cartilage tissue engineering.
In order to enhance the likelihood of bone healing process from fracture, currently bone graft combined with osteocytes is a new alternative in promoting new bone formation. Silica-based nanoparticles had been proven to induce intrinsic biology activity, especially in promoting bone-formation and differentiation of mesenchymal stem cells towards osteocytes. Previous studies had shown positive results of new bone tissue formation by incorporating SiO2 nanoparticles synthesized using TEOS. In this current study, the experiment was using micropatterning technique, with PDMS as the substrate, which was coated by SiO2 nanoparticle powder with the particle size distribution between 150nm-450nm. The effect of SiO2-coated PDMS on (Wharton’s jelly mesenchymal stem cells) hWJ-MSCs viability was evaluated using MTT cytotoxicity assay for 1, 3, 5, and 7 days, which was compared with two treatments: cells grown with SiO2 powder without patterns and cells on 96 well plate). The MTT assay result showed that SiO2-coated PDMS micropattern was non-toxic to the cells and had the highest increase in cell viability for 7 days in comparison to the two controls used.
This study aimed to determine the characteristics of scaffolds made of fibroin from Bombyx mori and spidroin from Argiope appensa in supporting the attachment and proliferation of HDF cells on the scaffolds. Thin-film scaffolds were made using the solvent casting technique, where the scaffold is an amalgamation of fibroin, spidroin, PVA, and glycerol. HDF cells were grown on DMEM medium with 10% FBS and 1% antibiotic-antimicotic. Characterization of the scaffolds was performed by using ATR-FTIR, swelling test, contact angle measurement, tensile test, biodegradation, MTT and SEM. The results of the ATR-FTIR analysis showed that the scaffolds contained fibroin, spidroin, PVA, and glycerol. Swelling and contact angle tests showed that all scaffold combinations were hydrophilic. Mechanical properties and in vitro biodegradation tests showed no significant difference among the scaffold combinations. MTT testing showed that all scaffolds could facilitate the attachment of fibroblasts and showed increased viability from day 1, 3, and 5. Scanning electron microscopy showed that the cells in the 70% fibroin and 10% spidroin scaffold had the best cell morphology and the best combination for potential application in skin tissue engineering.
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