Effect of High- and Low-Molecular-Weight Hyaluronic-Acid-Functionalized-AZ31 Mg and Ti Alloys on Proliferation and Differentiation of Osteoblast Cells

The quality of patient care has increased dramatically in recent years because of the development of lightweight orthopedic metal implants. The success of these orthopedic implants may be compromised by impaired cytocompatibility and osteointegration. Biomimetic surface engineering of metal implants using biomacromolecules including hyaluronic acid (HA) has been used an effective approach to provide conditions favorable for the growth of bone forming cells. To date, there have been limited studies on osteoblasts functions in response to metal substrates modified with the hyaluronic acid of different molecular weight for orthopedic applications. In this study, we evaluated the osteoblasts functions such as adhesion, proliferation, and differentiation in response to high- and low-molecular-weight HA (denoted as h-HA and l-HA, respectively) functionalized on Ti (h-HA-Ti and l-HA-Ti substrates, respectively) and corrosion-resistant silane coated-AZ31 Mg alloys (h-HA-AZ31 and l-HA-AZ31). The DNA quantification study showed that adhesion and proliferation of osteoblasts were significantly decreased by h-HA immobilized on Ti or AZ31 substrates when compared to low-molecular-weight counterpart over a period of 14 days. On the contrary, h-HA significantly increased the osteogenic differentiation of osteoblast over l-HA, as confirmed by the enhanced expression of ALP, total collagen, and mineralization of extracellular matrix. In particular, the h-HA-AZ31 substrates greatly enhanced the osteoblast differentiation among tested samples (l-HA-AZ31, l-HA-Ti, h-HA-Ti, and Ti alone), which is ascribed to the osteoinductive activity of h-HA, relatively up-regulated intracellular Ca²⁺ ([Ca²⁺]_i) and Mg²⁺ ([Mg²⁺]_i) concentrations as well as the alkalization of the cell culture medium. This study suggesting that HA of appropriate molecular weight can be successfully used to modify the surface of metal implants for orthopedic applications.