Effect of modified compound calcium phosphate cement on the differentiation and osteogenesis of bone mesenchymal stem cells.

BACKGROUND: The aim of this study is to evaluate the effect of self-invented compound calcium phosphate cement upon the proliferation and osteogenesis of bone mesenchymal stem cells (BMSCs). METHODS: Four groups including traditional calcium phosphate cement, modified calcium phosphate cement, modified calcium phosphate cement plus bone morphogenetic protein (BMP), and control groups were established. The cell proliferation curve was delineated by MTT. The activity of BMSCs to synthesize alkaline phosphatase (AKP) was evaluated. The growth and invasion of BMSCs were observed. The expression levels of aggrecan, collagen I, collagen II, AKP, and OSX messenger RNA (mRNA) were measured by using RT-PCR. RESULTS: Compared with other groups, the BMSCs in the modified calcium phosphate cement group presented with loose microstructure and the BMSCs closely attached to the vector margin. At 7 days after co-culture, the expression of AKP in the modified calcium phosphate cement plus BMP group was significantly upregulated compared with those in other groups. In the modified calcium phosphate cement group, the BMSCs properly proliferated on the surface of bone cement and invaded into the cement space. At 10 days, the expression levels of aggrecan, collagen I, collagen II, AKP, and OSX mRNA in the modified calcium phosphate cement and modified calcium phosphate cement plus BMP groups were significantly upregulated than those in other groups. CONCLUSIONS: Modified compound calcium phosphate cement possesses excellent biocompatibility and osteogenic induction ability. Loose microstructure and large pore size create a favorable environment for BMSCs proliferation and vascular invasion, as an ideal vector for releasing BMP cytokines to mediate the differentiation and osteogenesis of BMSCs.

Natural bone collagen scaffold combined with OP-1 for bone formation induction in vivo.

The scaffold is a key element to osteogenic tissue engineering as it provides a microenvironment for bone formation. Natural bone collagen scaffold (NBCS) is a novel biomaterial scaffold acid-extracted from organic human bone. The objective of this study was to characterize NBCS and evaluate the osteoconductivity of the scaffold, in combination with osteogenic protein-1 (OP-1), using a rabbit posteolateral lumbar fusion model. Thirty two rabbits were divided into 4 experimental groups, autograft, NBCS alone, OP-1 alone or NBCS combined with OP-1. Bone formation was evaluated by micro-CT, quantitative histological analysis, immunohistochemistry and semi-quantitative RT-PCR at 6 weeks postoperatively. By scanning electronic microscope, we showed that NBCS maintains a porous, interconnecting microarchitecture. Micro-CT analysis demonstrated that NBCS combined with OP-1 significantly induced (p < 0.01) bone formation at the fusion site as compared to control groups. This was confirmed by quantitative histological analysis which demonstrated that the NBCS combined with OP-1 significantly enhanced bone matrix area (17.7 mm(2)) (p < 0.05) and bone marrow cavity size (71.3 mm(2)) (p < 0.05) as compared to the controls. Immunohistochemical assessment and RT-PCR also demonstrated that NBCS combined with OP-1 enhanced type I collagen and osteonectin expression. Together, these results suggest that NBCS is an effective scaffold for osteogenesis, and combined with growth factors such as OP-1, possesses both osteoconductive and osteoinductive properties that are sufficient for bone regeneration.