Cell attachment and proliferation of osteoblast cell line (MG-63) on silk fibroin membrane for guided bone regeneration

Abstract

Purpose: The aim of this study is to verify the feasibility of silk fibroin, which is a typical fibrous protein produced by a variety of insects including the silkworm. Silk consists of two types of proteins, fibroin and sericin. Fibroin is the protein that forms the filaments of silkworm silk and it can be regenerated in various forms. These include membranes for guided bone regeneration by quantifying various biological responses such as cell attachment, cell viability, and cell proliferation of MG-63 osteoblast-like cells on a silk fibroin (SF) membrane.

Methods: Osteoblast cells (MG-63) were cultured on a silk fibroin membrane. The surface morphology and cell attachment were examined by scanning electron microscope (SEM) after 1, 5, and 7 days. Cell viability was determined by MTT (3-94,5-Dimethyulthiazol-2-yl)-2,5-Ddiphenyltetrazolium Bromide) assay after 1, 5, and 7. Cell proliferation was followed using DAPI fluorescence staining also at 1, 5, and 7.

Results: Optical microscopy revealed that MG-63 cells cultured on a silk fibroin (SF) membrane (group B) were able to multiply and proliferate around the SF membrane over a 7 day time period. Cell attachment to the surface of the SF membrane was examined for seven days using SEM. Comparison of the cell viability by the MTT assay between the cells grown on an SF membrane and those grown without attachment to an SF membrane (group A) showed increasing cell viability in both groups over time (p<0.05). While group A appeared to have greater viability over time, there was no statistically significant difference between the two groups (p>0.05). In contrast, DAPI staining for cell proliferation showed a statistically significant increase in group B over group A during the time of the study (p<0.05).

Conclusions: Silk fibroin membrane research is a study to complement the problems of currently used shielding materials and to show the possibilities of silk fibroin as a clinical alternative to the costly dental materials for guided bone regeneration (GBR).