Chondrogenic differentiation of mesenchymal stem cells (MSCs) is accurately regulated by essential transcription factors and signaling cascades. However, the precise mechanisms involved in this process still remain to be defined. MicroRNAs (miRNAs) regulate various biological processes by binding target mRNA to attenuate protein synthesis. To investigate the mechanisms for miRNAs-mediated regulation of chondrogenic differentiation, we identified that miR-145 was decreased during transforming growth factor beta 3 (TGF-β3)-induced chondrogenic differentiation of murine MSCs. Subsequently, dual-luciferase reporter gene assay data demonstrated that miR-145 targets a putative binding site in the 3′-UTR of SRY-related high mobility group-Box gene 9 (Sox9) gene, the key transcription factor for chondrogenesis. In addition, over-expression of miR-145 decreased expression of Sox9 only at protein levels and miR-145 inhibition significantly elevated Sox9 protein levels. Furthermore, over-expression of miR-145 decreased mRNA levels for three chondrogenic marker genes, type II collagen (Col2a1), aggrecan (Agc1), cartilage oligomeric matrix protein (COMP), type IX collagen (Col9a2) and type XI collagen (Col11a1) in C3H10T1/2 cells induced by TGF-β3, whereas anti-miR-145 inhibitor increased the expression of these chondrogenic marker genes. Thus, our studies demonstrated that miR-145 is a key negative regulator of chondrogenic differentiation by directly targeting Sox9 at early stage of chondrogenic differentiation.
Dural closure after the neurosurgery can prevent postoperative complications. Although many types of dural substitute have been developed, most of them lack functional and structural characteristics compared with the natural dura mater. In this study, we used electrospinning method to fabricate a multilayer scaffold to promote dural repair. The inner layer of the scaffold that faces the brain tissue is composed of poly-lactic acid (PLA) to reduce tissue adhesion. The middle layer of the scaffold is composed of poly-ɛ-caprolactone and PLA, which provides a watertight seal. The outer layer of the scaffold contains a large amount of collagen to promote cell attachment and proliferation. The results from in vitro study and an animal model have shown that this multilayer fibrous scaffold has sufficient mechanic strength and biochemical properties to enhance dural repair. Therefore, fabrication of scaffold with multiple functional and structural layers may provide a novel approach for tissue engineering.
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