A Minimal Common Osteochondrocytic Differentiation Medium for the Osteogenic and Chondrogenic Differentiation of Bone Marrow Stromal Cells in the Construction of Osteochondral Graft

Abstract

To regenerate the complex tissue such as bone-cartilage construct using tissue engineering approach, controllable differentiation of bone marrow stromal cells (BMSCs) into chondrogenic and osteogenic lineages is crucially important. This study proposes to test a minimum common osteochondrocytic differentiation medium (MCDM) formulated by including common soluble supplements (dexamethasone and ascorbic acid) used to induce chondrogenic and osteogenic differentiation. The MCDM coupled with supplemented growth factors was tested for its ability to differentiate BMSCs into osteogenic and chondrogenic lineages in both two-dimensional and three-dimensional culture systems. When transforming growth factor beta3 was added to MCDM, BMSCs differentiated to chondrocyte-like cells, evidenced by the expression of glycosaminoglycans and type II collagen, whereas osteogenic differentiation was induced by supplementing osteogenic protein-1, resulting in detectable expression of osteopontin and osteocalcin. These chondrogenic and osteogenic differentiation markers were significantly enhanced in the three-dimensional cultures compared to the two-dimensional monolayer cultures. The results achieved in this study lay a foundation for future development of osteochondral graft, which could be engineered from bilayered scaffold with spatially loaded growth factors to control BMSC differentiation.

Serum-specific stimulation of proliferation and mineralization of fish bone-derived cells

“…Changes in expression of other bone‐related genes did not provide clear evidence of their participation in the mineralogenic effect of GSS. Expression of osteocalcin 1 (OC1), a Gla‐containing protein associated with tissue ossification (Ducy et al., 1996; Hunter et al., 1996) and a marker of osteoblast differentiation (Nieden et al., 2005; Li et al., 2009), was apparently significantly reduced in mineralizing GSS‐treated VSa13 cells but was found at levels close to qPCR detection limits in all conditions. Previous expression data showing its absence in VSa13 cells (Pombinho et al., 2004) suggest that the observed decrease in osteocalcin expression in GSS‐treated cells may be negligible.…”

Bioactive inorganic‐materials/alginate composite microspheres with controllable drug‐delivery ability

“…Isolation and culture of BMSCs was conducted following previously published protocols 23. BMSCs (1 × 10 5 /well) were seeded on 96‐well template and allowed to adhere to the template for 3 h. Then, microspheres were added to the template.…”

Biphasic Nanofibrous Constructs with Seeded Cell Layers for Osteochondral Repair

“…The chondrogenic medium consists of the normal medium (DMEM + 100 U/mL penicillin/100 mg/mL streptomycin + 10% FBS) + 1% insulintransferrin-selenium (PAA Laboratories, Inc.) + 37.5 mg/mL ascorbic acid + 100 nM dexamethasone + 10 ng/mL transforming growth factor-b1 (PeproTech), and the osteogenic medium is made of the normal medium (a-MEM + 100 U/mL penicillin/100 mg/mL streptomycin + 10% FBS) + 50 mg ascorbic acid + 10 mM b-glycerophosphate + 100 nM dexamethasone. 18 Within each medium, the cell-nanofiber constructs were cultured for up to 14 days, and the medium was refreshed every 2-3 days. In case of cells without having undergone specific differentiation, the normal medium (as described above) was used for each type of cells.…”