Osteogenesis of adipose-derived stem cells on three-dimensional, macroporous gelatinhyaluronic acid cryogels

摘要

Macroporous sponge-like gelatinhyaluronic acid (GlHA) scaffolds cross-linked by EDC were produced using cryogelation technology, which allows for the preparation of highly porous scaffolds without compromising their mechanical properties, and is a more cost-efficient process than freeze drying. The aim of this study is to evaluate the osteogenic potential of porcine adipose-derived stem cells (PADSCs) in GIHA cryogel. Method. The character of the GIHA cryogel was evaluated. The pore size and the microstructure were observed using scanning electron microscope (SEM). The swelling ratio was measured. The PADSCs were harvested and isolated from pig inguinal area. Then, the GIHA cryogel was seeded with PADSCs. The cryogel/ASCs mixture was cultured in osteogenic medium for 0, 3, 7, 14, and 21 days. The cell proliferation was measured by MTS. The RT-PCR of specific osteogenic gene expression such as osteocalcin (OC), RUNX2 was used to assess the osteogenic ability. The SEM was used to observe the interaction between scaffold and cells. Energy dispersive spectrum (EDS) was used to analyze the mineralization around cells. Results. The pore size was variable between 200 and 369 μm. The swelling ratio was around 8.67 ± 1.669%. The cell proliferation was increasing along with the increase of induction periods. The expression of early gene of RUNX2 and late gene of OC mean that the PADSCs were differentiated well into osteoblasts within the cryogels. The SEM detailed that the PADSCs cell can proliferate well in the pore of GIHA scaffold. The EDS also demonstrated the mineralization of PADSCs in GIHA scaffold after induction. Conclusions. To conclude, the PADSCs can proliferate and differentiate well into osteoblasts in the three-dimensional, porous, GIHA cryogel.