Attachment, spreading, and proliferation of Wharton’s jelly mesenchymal stem cells on scaffold combination of silk fibroin and Argiope appensa silk spidroin

In tissue engineering, the structure and material composition of scaffold play important roles in facilitating cell attachment and spreading, as the first important step of cell survival. Focal adhesion between scaffold and cell determines physical contact that initiates biological processes in the cell. Aims of this research were (1) to fabricate a scaffold with the combination of silk fibroin (SF) from Bombyx mori and silk spidroin (SS) from Argiope appensa, (2) to observe the effect of SS addition on attachment and spreading of hWJ-MSCs (Human Wharton's Jelly Mesenchymal Stem Cells); and (3) analyzing the biocompatibility and optimization of the scaffold composition on cell growth. This current study began with the fabrication of a scaffold using salt-leaching method with several variations of composition and NaCl as porogen with size ranged from 470 - 530 µm. hWJ-MSCs morphology and attachment to the scaffold surface were observed using Scanning Electron Microscope (SEM). Cell proliferation on the silk scaffolds was analyzed using MTT, while cell attachment observed through the expression of integrin β1 using confocal microscopy and quantified using ImageJ software. Observation of cell morphology on the scaffold showed that hWJ-MSCs on the SF 100% scaffold had a round shape that showed weak cell attachment, whereas the hWJ-MSCs on the scaffold with silk spidroin was elongated and flattened and spread well. The cells had higher cell viability on the scaffold with 10% SS for over 7 days in comparison to SF 100% scaffold. Integrin expression also showed that scaffold with spidroin silk could support cell adhesion and proliferation better than SF 100%. The results of the study were concluded as such; (1) the porous scaffold as a combination of Bombyx mori fibroin and A. appensa spidroin with a concentration of 100%, SF 90% + SS 10% was successfully fabricated, (2) the addition of silk spidroin increased cell attachment and spreading, and (3) based on the MTT assay cells grown on SF 90% + SS 10% scaffold had higher cell viability for 7 days in comparison to 100% SF scaffold.