Microfabrication of Gelatin Methacrylate/Hydroxyapatite Composites by Utilizing Alternate Soaking Process.

To construct a complex three-dimensional (3D) structure mimicking bone microstructure, hydrogel models of polymerized gelatin methacrylate (pGelMA) were fabricated by using stereolithography and modified with hydroxyapatite (HAp) via an alternate soaking process (ASP) using a solution of calcium and phosphate ions. Fabricated pGelMA line models whose widths were designed as 100, 300, and 600 µm were modified with HAp by ASP by changing the immersion time and number of cycles. After ASP, all of the line models with widths of 100, 300, and 600 µm were successfully modified with HAp, and large amounts of HAp were covered with the fabricated models by increasing both the immersion time and the number of cycles in ASP. HAp was observed near the surface of the line model with a width of 600 µm after ASP at an immersion time of 10 s, while the entire model was modified with HAp using ASPs for longer immersion times. The adhesion and spread of mesenchymal stem cells (MSCs) on the pGelMA-HAp discs depended on the ASP conditions. Moreover, the HAp modification of 3D pyramid models without alteration of the microstructure was also conducted. This two-step fabrication method of first fabricating frameworks of hydrogel models by stereolithography and subsequently modifying the fabricated models with HAp will lead to the development of 3D cell culture systems to support bone grafts or to create biological niches, such as artificial bone marrow.

Surface modification of silica nonwoven fabrics for osteogenesis of bone marrow-derived mesenchymal stem cells.

Silica nonwoven fabrics (SNFs) with high mechanical strength and porosity are known to exhibit high cell proliferation and osteogenic differentiation potential of mesenchymal stem cells (MSCs) by morphologically mimicking the extracellular matrix (ECM). To further improve the osteoinductive ability of SNFs, it could be effective to increase the interaction between MSCs and ECM components because exogenous ECM components seem to modulate the fate of MSCs differentiation. In this study, we developed immobilization methods for ECM components, such as collagen, fibronectin, and chondroitin sulphate C on SNFs, to improve cell-matrix interactions and examined their suitability for bone tissue regeneration. Collagen and fibronectin were immobilized via physical adsorption and chondroitin sulphate C was also immobilized by the layer-by-layer method combined with chitosan on SNF surfaces to maintain the high porosity of SNFs. The treated SNFs were characterized using scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. In osteogenic differentiation culture, modified SNFs showed significantly increased expression of osteogenic differentiation marker genes compared to unmodified SNFs. These results suggest that the present methods improve cell-matrix interactions and enhance the cellular functions of MSCs. We are convinced that these simple modification techniques for ECM components are effective in functionalizing various 3D fabric scaffolds possessing hydrophilic groups.