Cross-linked gelatin microsphere-based scaffolds as a delivery vehicle of MC3T3-E1 cells: in vitro and in vivo evaluation

摘要

Scaffolding plays a crucial role in bone tissue engineering by not only providing interfaces for cell adhesion, proliferation, and differentiation but also guiding neotissue formation. For this purpose, microspheres (MSs) are being increasingly used alone or in combination with other scaffolds. However, few researchers have used MSs to prepare 3D scaffolds by culture with delivered cells. In this study, we have developed covalent cross-linked gelatin MSs (ccG-MSs) (average diameter = 100-300 mu m) to load mouse osteoblast MC3T3-E1 cells, which exhibit attachment and spreading on surfaces of ccG-MSs after co-culture. Significantly, the ccG-MSs can be integrated into a macroscopic construct with MC3T3-E1 cells after 5 days of cultivation. The MC3T3-E1 cells within ccG-MSs constructs show a higher viability and proliferation activity than those in the micro-cavitary gelatin gel (MCG) constructs. Calcium deposition, alkaline phosphatase activity as well as osteocalcin secretion within both ccG-MSs and MCG constructs have been evaluated in vitro and in vivo, respectively. Compared to MCG scaffolds, ccG-MS-based scaffolds can provide better cellular microenvironments for cell proliferation and osteogenic differentiation. Our findings will lay the foundation for understanding cellular behaviors in MS-based 3D constructs and help in designing MS-based bone tissue engineering scaffolds.