Osteoblastic differentiation of cultured rat bone marrow cells on hydroxyapatite with different surface topography

Abstract

Hydroxyapatite (HA) has been used in orthopedic, dental, and maxillofacial surgery as a bone substitute. Objective. The aim of this investigation was to study the effect of surface topography produced by the presence of microporosity on the response of the rat bone marrow cells, evaluating: cell attachment, proliferation, total protein content, alkaline phosphatase (ALP) activity, and bone-like nodule formation. Methods. Cells were cultured on HA discs manufactured by a combination of uniaxial powder pressing and different sintering conditions, with different percentage of microporosity (<5%-HA5, 15%-HA15, and 30%-HA30). For attachment evaluation, cells were cultured for 2 h. Proliferation was evaluated after 7 and 14 days. After 14 days, total protein content and ALP activity were measured. For bone-like nodule formation, cells were cultured for 21 days. Data were compared by ANOVA and Duncan's multiple range test when appropriate. Results. Cell attachment was not affected by surface topography ( p=0.37). Proliferation ( p=0.001), total protein content ( p=0.039), ALP activity ( p=0.050), and bone-like nodule formation ( p=0.00001) were all significantly decreased by the most irregular surface (HA30). Significance. These results suggest that initial cell events were not affected by the surface topography of the HA. However, intermediary and final events such as proliferation, protein synthesis, ALP activity, and bone-like nodule formation favored surfaces with a more regular topography, such as that presents in HA with 15% or less of microporosity.