In vitro triple cultures of human primary osteoblasts, osteocytes and osteoclasts can potentially help to analyze the effect of drugs and degradation products of biomaterials as a model for native bone tissue. In the present study, degradation products of Magnesium (Mg), which has been successfully applied in the biomedical field, were studied with respect to their impact on bone cell morphology and differentiation both in osteocyte single cultures and in the triple culture model. Fluorescence microscopic and gene expression analysis, analysis of osteoclast- and osteoblast-specific enzyme activities as well as osteocalcin protein expression were performed separately for the three cell types after cultivation in triple culture in the presence of extracts, containing 5 and 10 mM Mg2+. All three cell species were viable in the presence of the extracts and did not show morphological changes compared to the Mg-free control. Osteoblasts and osteoclasts did not show significant changes in gene expression of ALPL, BSPII, osteocalcin, TRAP, CTSK and CA2. Likewise on protein level, no significant changes in ALP-, TRAP-, CTSK- and CAII activities were detected. Osteocytes showed a significant downregulation of MEPE, which codes for a protein playing an important role in regulation of phosphate homeostasis by osteocytes. This study is the first to analyze the effects of Mg degradation products on primary osteocytes in vitro, both in single and triple culture. Even if promoting effects on the three examined bone cell species were not found in the applied triple culture setup, it was shown, that Mg degradation products do not interfere with the activity of osteoblasts, osteoclasts and osteocytes in vitro.
Keywords: Co-culture; Degradable magnesium; In vitro bone model; Metallic implant; Osteoblast; Osteoclast; Osteocyte; Triple culture.
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