ABSTRACT
Scaffold topography and culture medium conditions for human wharton's jelly mesenchymal stem cells (hWJ-MSC) are critical components of the approach to nucleus pulposus (NP) tissue engineering. Aim: To evaluate the silk fibroin (SF) scaffold topography analysis (optimal thickness and pore diameter) and to determine culture medium conditions for the growth and differentiation of hWJ-MSC. Method: hWJ-MSCs were seeded into different thicknesses and pore size diameters and grown in different concentrations of glucose, platelet rich plasma (PRP) and oxygen. The cell-seeded scaffold was evaluated for cell attachment, growth and differentiation potency. Results & discussion: The results indicated that SF scaffold with a minimum thickness 3.5 mm and pore diameter of 500 µm with cells cultured under low glucose, 10% PRP and normoxia conditions induced the growth and differentiation of hWJ-MSCs, indicated by the accumulation of glycosaminoglycans content and the presence of type II collagen, as markers of NP-like cells.
ABSTRACT
The data showed how gelatin hydrogel and silk fibroin scaffolds could facilitate the growth of human Mesenchymal Stem Cells (hMSC). Gelatin hydrogel and silk fibroin are biodegradable materials. Gelatin hydrogel already has many uses in the medical field, especially in tissue engineering, but silk fibroin scaffold, which is made from the cocoon of silkworm by salt leaching, its role in facilitating growth of hMSC still needs to be proven. Data was obtained by characterization of hMSC, then growing hMSC on silk fibroin scaffolds with pore sizes of ±500 µm and ±900 µm and on gelatin hydrogel scaffolds as control. Testing was performed by counting cell growth on days 1, 3, 5, 7 and 14 with the MTT cytotoxicity assay protocol. The morphology of hMSC that grew on gelatin and silk fibroin scaffolds was observed with a Scanning Electron Microscope (SEM) on day 3. Characterization of the hMSC showed that it fulfilled the requirements of the International Society for Cellular Therapy (ISCT). The water content of the gelatin hydrogel scaffold was higher than the silk fibroin scaffold. Biocompatibility testing showed that the gelatin hydrogel scaffold could support cell growth until day 7, then decreased on day 14 compared to the silk fibroin scaffold based on absorbance on the MTT cytotoxicity assay, while growth on silk fibroin scaffold with pore size 833 ± 147 µm was consistently higher than on pore size 462 ± 66 µm from day 1 to day 14. Cell binding to the silk fibroin was proven from SEM observation.
