The Effect of Various Surface Treatments of Ti6Al4V on the Growth and Osteogenic Differentiation of Adipose Tissue-Derived Stem Cells

Abstract

The physical and chemical properties of the material surface, especially its roughness and wettability, have a crucial effect on the adhesion, proliferation, and differentiation of cells. The aim of this study is to select the most appropriate surface modifications of Ti6Al4V implants for pre-colonization of the implants with adipose tissue-derived stem cells (ASCs) in order to improve their osseointegration. We compared the adhesion, growth, and osteogenic differentiation of rat ASCs on Ti6Al4V samples modified by methods commonly used for preparing clinically used titanium-based implants, namely polishing (PL), coating with diamond-like carbon (DLC), brushing (BR), anodizing (AND), and blasting (BL). The material surface roughness, measured by the Ra and Rq parameters, increased in the following order: PL < DLC ˂ BR ˂ AND ˂ BL. The water drop contact angle was in the range of 60–74°, with the exception of the DLC-coated samples, where it was only 38°. The cell number, morphology, mitochondrial activity, relative fluorescence intensity of osteogenic markers RUNX2, type 1 collagen, and osteopontin, the calcium consumption by the cells and the alkaline phosphatase activity depended on the surface roughness rather than on the surface wettability of the materials. Materials with a surface roughness of several tens of nanometers (Ra 60–70 nm), i.e., the BR and AND samples, supported a satisfactory level of cell proliferation. At the same time, they achieved the highest level of osteogenic cell differentiation. These surface modifications therefore seem to be most suitable for pre-colonization of Ti6Al4V implants with stem cells pre-differentiated toward osteoblasts, and then for implanting them into the bone tissue.