Egg shell membrane – a potential natural scaffold for human meniscal tissue engineering: an in vitro study

Abstract

In the present study, natural egg shell membrane (ESM), harvested from locally available single comb white leghorn hen eggs was investigated for its ability to support adhesion and proliferation of human meniscal cells. The harvested ESM was subjected to moist heat (autoclaving) and compared with raw egg shell membrane (RESM) for meniscal cell growth. RESM and autoclaved egg shell membrane (AESM) were characterized using suitable techniques like field emission scanning electron microscopy (FESEM), solid surface zeta potential, Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and biodegradability in trypsin and phosphate buffered saline (PBS). From the characterization studies, it was evident that autoclaving resulted in surface modification of RESM. RESM was cationic in nature and was covered with a mucilaginous coating on the surface. However, AESM showed almost neutral surface charge without any mucilaginous coating. The AESM showed increased resistance to biodegradation when compared to RESM. The primary human meniscal cells, seeded onto the fibrous side of both ESM showed increased cell adhesion and proliferation in AESM than RESM. Extracellular matrix (collagen and glycosaminoglycan) secretion into the external medium by the adhered cells was higher with AESM than RESM. Cell attachment, DNA content on the scaffold, cell proliferation index, cytotoxicity and biodegradation studies also confirmed that AESM scaffolds supports better cell attachment and growth of meniscal cells when compared to RESM. Hence, AESM can be a potential and interesting natural scaffold matrix for meniscal tissue engineering.