Comparative In Vitro Osteoinductivity Study of CaP Ceramics (HA, α-TCP, β-TCP) using 10T1/2 Cells with Different Controls and Possible Correlations with Other Systems

Juliana Tsz Yan Lee, Kefeng Wang, Wai Hung TSANG, King Lau Chow
DOI: 10.4236/jbnb.2011.22021                

Abstract

In this study, we use a pluripotent mesenchymal stem cell (MSC) model, C3H/10T1/2, to evaluate three calcium phos-phate (CaP) materials, namely the hydroxyapatite (HA), α-tricalcium phosphate (α-TCP) and β-tricalcium phosphate (β-TCP). 10T1/2 cell was chosen as it has advantages over its counterparts in terms of ease of maintenance, free of ethical concerns and also more reproducible results. ALP enzymatic assay, RT-qPCR, DAPI staining and SEM were employed to assess the osteoinductivity of these materials. A good reference material which also acts as a scientific control is necessary for comparisons of results from different experimental batches and hence other materials such as titanium, Nunclon plastic surface, BD Falcon plastic surface and gold coated porous HA were also tested. The results show that ceramics induce a more sustained osteo-differentiation state as compared with plastics. Inductivity was found to be acting in descending order of strength with HA > β-TCP > α-TCP, which is reversed in terms of their impact on proliferation rate (HA < TCP). This is also consistent with the results observed in SBF study in terms of calcium phosphate precipitate area coverage (HA > TCP) and in vivo osteoinductivity in terms of incidence and quality of bone described previously (HA > β-TCP > α-TCP). These confirm the suitability of using 10T1/2 cells in cell culture assay of osteoinductivity.

Keywords

Bone Tissue Engineering; Stem Cell; Calcium Phosphate; Hydroxyapatite; Gene Expression; Osteo-Inductivity; In Vitro Test; Pcr (Polymerase Chain Reaction); Sem (Scanning Electron Microscopy)

Conflicts of Interest

The authors declare no conflicts of interest.

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