Previous experiments showed that microgrooved substrate surfaces can influence the in vitro behavior of osteoblast-like rat bone marrow (RBM) cells. Cellular morphology and matrix deposition can be directed by substrate chemistry and topography. RBM cells cultured on poly-l-lactic acid (PLA) exhibit increased mineralization and alkaline phosphatase activity compared to polystyrene substrates. Consequently, the purpose of the present in vitro study is to further evaluate the behavior of RBM cells on microgrooved (groove depth 0.5 to 1.5 microns, groove and ridge width 1 to 10 microns) polystyrene (PS) and PLA surfaces. Besides grooved, also smooth control surfaces were made. Our results confirmed that microtextured surfaces are capable of influencing the behavior of the osteoblast-like RBM cells in vitro. Microtopography did not influence the RBM proliferation rate, or cellular actin organization. However, confocal laser scanning microscopy showed that cellular attachment is dependent on the applied material. Also clear differences were found between textured PS and PLA, with regard to the calcium content. We therefore conclude, that the application of microtextures could possibly influence the bone regeneration around biodegradable PLA devices.
The aim of this study was to evaluate the osteogenic properties of magnetron sputtered dicalcium pyrophaosphate (DCPP) and hydroxylapatite (HA) coatings. Therefore, DCPP and HA coatings were deposited on grit-blasted titanium discs. The substrates were used as-prepared or received an additional heat treatment which changed the amorphous coating structure to a crystalline structure. Subsequently, rat bone marrow stromal cells were cultured for 1-24 days on the various substrates. DNA and alkaline phosphatase activity was determined after 1, 3, 5, 8, and 12 days of incubation. Osteocalcin expression was evaluated after 8, 12, 16, and 24 days of incubation. Scanning electron microscopical analysis of cell morphology and coating characteristics was done after 8 and 16 days of incubation. All assays were done in duplicate and in each assay all specimens were present in fourfold. Results demonstrated that the cells did not proliferate and differentiate on all amorphous coatings. SEM revealed that the amorphous coatings showed significant dissolution. On the crystalline DCPP and HA coatings an increase in DNA and alkaline phosphatase activity was seen starting at day 8 of incubation. Osteocalcin expression on the crystalline coatings started to increase at day 16 of incubation. SEM showed that the growth and differentiation of the cells was associated with extensive collagen fiber formation and surface mineralization in the form of globular accretions. Further, statistical testing revealed that proliferation and differentiation of the rat bone marrow stromal cells started significantly earlier on the crystalline HA coatings than that on the crystalline DCPP coatings. These results demonstrate that the rat bone marrow stromal cells proliferated and differentiated only on crystalline magnetron sputtered DCPP as well as HA coatings, which warrants the further in vivo analysis of the bone healing supporting properties of these coatings.
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