Background
Most of the natural extracellular matrix (ECM) is a three-dimensional (3D) network structure of micro/nanofibers for cell adhesion and growth of 3D. Electrospun fibers distinctive mimicked 2D ECM, however, it is impossible to simulate 3D ECM because of longitudinal collapse of continuous micro/nanofibers. Herein, 3D electrospun micro/nano-fibrous sponge was fabricated via electrospinning, homogenization, shaping and thermal crosslinking for 3D tissue regeneration of cells and vascular.
Results
Fibrous sponge exhibited high porosity, water absorption and compression resilience and no chemical crosslinked agent was used in preparation process. In vitro studies showed that the 3D short fiber sponge provided an oxygen-rich environment for cell growth, which was conducive to the 3D proliferation and growth of HUVECs, stimulated the expression of VEGF, and well promoted the vascularization of HUVECs. In vivo studies showed that the 3D short fiber sponges had a good 3D adhesion to the chronic wound of diabetes in rats. Furthermore, 3D short fibrous sponges were better than 2D micro/nanofiber membranes in promoting the repair of diabetic full-thickness skin defects including wound healing, hair follicle regeneration, angiogenesis, collagen secretion.
Conclusion
Therefore, electrospun short fibrous sponges are special candidates for mimicking the 3D ECM and promoting 3D regeneration of tissue.
Graphic Abstract
Background:
Osteosarcoma (OS) is one of the most common malignant tumors of bone, and microRNAs (miRNAs/miRs) serve critical roles in the progression of human OS. The aim of the present study was to investigate the role of miR-495-3p in OS.
Methods:
The expression of miR-495-3p in OS tissues and adjacent tissues from 30 patients was measured by reverse
transcription-quantitative PCR (RT-qPCR). Human OS cell lines (U-2 OS, MG-63 and Saos-2 cells) and normal osteoplastic cells (hFoB 1.19 cells) were employed to perform the further analysis. The cell proliferation ability of MG-63 cells was measured by Cell Counting Kit-8 assay and colony formation assay. In addition, cell invasion and migration were evaluated by Transwell and scratch wound healing assays, respectively. Flow cytometry was applied to assess cell apoptosis and the cell cycle. Moreover, RT-qPCR and Western blotting were performed to measure mRNA and protein expression. A luciferase reporter assay was used to verify the target gene of miR-495-3p. Furthermore, a xenograft OS model was made to evaluate the effect of miR-495-3p in vivo.
Results:
The results revealed that miR-495-3p was downregulated in the OS tissues and GBM cell lines. Additionally, miR-495-3p overexpression suppressed the proliferation, migration and invasion of MG-63 cells. Simultaneously, cell apoptosis was promoted, accompanied by cell cycle arrest, after transfecting with miR-495-3p mimics. In addition, the expression levels of cell apoptosis-related proteins were increased, whereas proteins of the cell cycle were decreased. Importantly, C1q/TNF-related protein 3 (CTRP3) was confirmed as a direct target of miR-495-3p. A xenograft tumor model was employed to verify the effects of miR-495-3p on OS.
Conclusion:
On the basis of these results, we conclude that miR-495-3p overexpression inhibited cell proliferation, migration and invasion by downregulating CTRP3. Therefore, miR-495-3p may act as a tumor suppressor and an underlying target for OS treatment.
Human embryonic stem cells (ESCs) can differentiate into endothelial cells in response to stimuli from extracellular cytokines. Transforming growth factor (TGF)-β1 signaling is involved in stem cell renewal and vascular development. Previously, human ESCs were isolated from inner cell mass and a stable ESC line was developed. In the present study, the effects of extracellular TGF-β1 were investigated on human ESC-derived embryoid bodies (EB) in suspension. The structures of the EBs were analyzed with light and electron microscopy, while the cellular composition of the EBs was examined via the expression levels of specific markers. Vascular-like tubular structures and cardiomyocyte-like beating cells were observed in the EBs at day 3 and 8, respectively. The frequencies of vascular-like structures and beating cells in the TGF-β1 treated group were significantly higher compared with the control group (84.31 vs. 12.77%; P<0.001; 37.25 vs. 8.51%; P<0.001, respectively). Electron microscopy revealed the presence of lumens and gap junctions in the sections of the tubular structures. Semiquantitative polymerase chain reaction revealed elevated expression levels of CD31 and fetal liver kinase-1 in EBs cultured with TGF-β1. In addition, extensive staining of von Willebrand factor was observed in the vascular-like structures of TGF-β1-treated EBs. Therefore, the results of the present study may aid the understanding of the underlying mechanisms of human ESC differentiation and improve the methods of propagating specific cell types for the clinical therapy of cardiovascular diseases.
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