Abstract
Micro- and nano-patterning/modification are emerging strategies to improve surfaces properties that may influence critically cells adherence and differentiation. Aim of this work was to study the in vitro biological reactivity of human bone marrow mesenchymal stem cells (hBMSCs) to a nanostructured titanium dioxide (TiO2) surface in comparison to a coverglass (Glass) in two different culture conditions: with (osteogenic medium (OM)) and without (proliferative medium (PM)) osteogenic factors. To evaluate cell adhesion, hBMSCs phosphorylated focal adhesion kinase (pFAK) foci were analyzed by confocal laser scanning microscopy (CLSM) at 24 h: the TiO2 surface showed a higher number of pFAK foci with respect to Glass. The hBMSCs differentiation to osteoblasts was evaluated in both PM and OM culture conditions by enzyme-linked immunosorbent assay (ELISA), CLSM and real-time quantitative reverse transcription PCR (qRT-PCR) at 28 days. In comparison with Glass, TiO2 surface in combination with OM conditions increased the content of extracellular bone proteins, calcium deposition and alkaline phosphatase activity. The qRT-PCR analysis revealed, both in PM and OM, that TiO2 surface increased at seven and 28 days the expression of osteogenic genes. All together, these results demonstrate the capability of TiO2 nanostructured surface to promote hBMSCs osteoblast differentiation and its potentiality in biomedical applications.
Highlights
Bone tissue engineering is a strategy to replace autologous or heterologous bone grafts with an artificial material that mimics the bone structure [1,2,3]
A bone graft substitute may follow a series of indication and should share a higher number of properties, such as a good material that allows a structural framework for bone growth and the good stem cell to produce new bone [4,5], good cell attachment and the absence of immune reactions
The surface of the Glass and of nanostructured TiO2 were different at scanning electron microscopy (SEM) (Figure 1): a uniform and particulate structure of the clusters, with diameter under 100 nm of dimension was observed for the TiO2 surface (Figure 1D–F) but not for the Glass (Figure 1A–C)
Summary
Bone tissue engineering is a strategy to replace autologous or heterologous bone grafts with an artificial material (scaffold) that mimics the bone structure [1,2,3]. The subsequent step is the production of new bone by the newly formed osteoblasts [6,7]. To properly allow this last event, materials should be resorbable. The resorption induced by osteoclasts may be counteracted by the production of new bone This gradual process takes a duration in the order of weeks/months and should ideally lead to the complete dissolution of the scaffold. It should consent to cell adhesion and promote cell growth and differentiation It should facilitate extracellular matrix regeneration and cell spatial distribution through the scaffold to mimic the physiological bone architecture [9]
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