Layer-by-layer (LBL) self-assembly technique has been proved to be a highly effective method to immobilize the main components of the extracellular matrix such as collagen and hyaluronic acid on titanium-based implants and form a polyelectrolyte multilayer (PEM) film by electrostatic interaction. However, the formed PEM film is unstable in the physiological environment and affects the long-time effectiveness of PEM film. In this study, a modified LBL technology has been developed to fabricate a stable collagen/hyaluronic acid (Col/HA) PEM film on titanium coating (TC) by introducing covalent immobilization. Scanning electron microscopy, diffuse reflectance Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy were used to characterize the PEM film. Results of Sirius red staining demonstrated that the chemical stability of PEM film was greatly improved by covalent crosslinking. Cell culture assays further illustrated that the functions of human mesenchymal stem cells, such as attachment, spreading, proliferation and differentiation, were obviously enhanced by the covalently immobilized Col/ HA PEM on TCs compared with the absorbed Col/HA PEM. The improved stability and biological properties of the Col/HA PEM covalently immobilized TC may be beneficial to the early osseointegration of the implants.
Immune systems play pivotal roles in determining the in vivo osseointegration of bone implants. While much evidence has shown that hierarchical implant surfaces can exhibit excellent osteogenic ability, their immune response has not been well elucidated, which will preclude accurate knowledge of their osseointegration performance. In this study, the immunomodulatory properties of a hierarchical macropore/nanosurface as well as the detailed mechanism was investigated. Macrophages were found to switch to M2 phenotype on the hierarchical surface, and decreased levels of inflammatory gene expression as well as increased expression of anti-inflammatory genes were endowed, which were probably regulated by the decisive role of cytoskeleton tension induced by specific cell shape. In addition, enhanced osteogenic differentiation of bone marrow mesenchymal stem cells and angiogenesis of human umbilical vein endothelial cells could be observed when stimulated by a RAW cells/hierarchical surface conditioned medium, which were probably due to increased expression of BMP-2 and VEGF of RAW cells, respectively. These findings give comprehensive knowledge into detailed mechanism of the immunomodulatory behavior of the hierarchical surface, which will also provide insight into the surface design of advanced bone biomaterials with satisfactory immunomodulation properties.
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