Osteogenesis‐Inducing Chemical Cues Enhance the Mechanosensitivity of Human Mesenchymal Stem Cells for Osteogenic Differentiation on a Microtopographically Patterned Surface

Mechanical cues are widely used for regulating cell behavior because of their overarching, extensive, and non‐invasive advantages. However, unlike chemical cues, mechanical cues are not efficient enough to determine cell fate independently and improving the mechanosensitivity of cells is rather challenging. In this study, the combined effect of chemical and mechanical cues on the osteogenic differentiation of human mesenchymal stem cells is examined. These results show that chemical cues such as the presence of an osteogenic medium, induce cells to secrete more collagen, and induce integrin for recruiting focal adhesion proteins that mature and cascade a series of events with the help of the mechanical force of the scaffold material. High‐resolution, highly ordered hollow‐micro‐frustum‐arrays using double‐layer lithography, combined with modified methacrylate gelatin loaded with pre‐defined soluble chemicals to provide both chemical and mechanical cues to cells. This approach ultimately facilitates the achievement of cellular osteodifferentiation and enhances bone repair efficiency in a model of femoral fracture in vivo in mice. Moreover, the results also reveal these pivotal roles of Integrin α2/Focal adhesion kinase/Ras homolog gene family member A/Large Tumor Suppressor 1/Yes‐associated protein in human mesenchymal stem cells osteogenic differentiation both in vitro and in vivo. Overall, these results show that chemical cues enhance the microtopographical sensitivity of cells. Chemical cues enhance the mechanical sensitivity of mesenchymal stem cells through Integrin α2/Focal adhesion kinase/Ras homolog gene family member A/Large Tumor Suppressor 1/Yes‐associated protein pathway. Further, double‐layer lithography and anisotropic etching is used to fabricate high‐resolution, highly ordered 3D hollow‐micro‐frustum‐arrays, combined with modified methacrylate gelatin loaded with pre‐defined soluble chemicals to provide both chemical and mechanical cues to cells to achieve osteogenic differentiation.