Kurzfassung

3D plotting and bioactive surface functionalisation are investigated with emphasis on the in vitro effect of Bone Sialoprotein (BSP) on primary human osteoblasts (hOBs). Our primary objective is to demonstrate the BSP influence on the expression of distinctive osteoblast markers in hOBs. Secondary objectives include the examination of the calcium phosphate cement (CPC) scaffolds surface and its binding properties in regard to BSP-coating as well as investigations of cell viability...3D plotting and bioactive surface functionalisation are investigated with emphasis on the in vitro effect of Bone Sialoprotein (BSP) on primary human osteoblasts (hOBs). Our primary objective is to demonstrate the BSP influence on the expression of distinctive osteoblast markers in hOBs. Secondary objectives include the examination of the calcium phosphate cement (CPC) scaffolds´ surface and its binding properties in regard to BSP-coating as well as investigations of cell viability and proliferation. 3D-plotted CPC scaffolds are coated with BSP via physisorption. hOBs are seeded on the coated scaffolds, followed by cell viability measurements for several days, gene expression analysis via qRT-PCR and visualisation through fluorescence microscopy. Physisorption is an effective method for immobilizing BSP on these scaffolds. Coating with higher BSP concentrations leads to higher residual BSP amounts on the bone substitutes. In the hOB gene expression analysis, coating with lower BSP concentrations (50 µg/ml) resulted in a significant decrease of ALP and SPARC expression, whereas higher BSP concentrations (200 μg/ml) showed an tendency towards increased gene marker expression, particularly of Runx2, SP7 and OPN. Enhanced cell viability was observed on BSP coated scaffolds on day 3. Adhesion characteristics of hOBs remained unaltered, whereas hOBs developed a polygonal shape and formed a meshwork under the influence of BSP. The strength of the above-mentioned effects of BSP-coated scaffolds in vivo is unknown, and future work is focusing on bone ingrowth and vascularisation in vivo.
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