Micropatterned Nanofibrous Membrane Promotes Wound Healing Via The Modulating LncRNA-XIST/miR-150-5p/COL1A1 Pathway

Objectives:Micropatterned nanofibrous membranes are widely used in tissue-related processes such as wound healing.The proliferation,migration and extracellular matrix formation of fibroblasts play a key role in wound healing.However,whether the micropatterned nanofiber membrane can effectively promote the proliferation,migration,and synthesis of extracellular matrix of fibroblasts and the specific mechanism need to be further studied.Methods:The surface morphology and structure of the fiber membrane were observed by optical microscope.The wettability and mechanical properties of the fiber membrane were tested.In vivo experiments were performed to compare the difference of surface wound area,epithelial cell thickness and scar between micropatterned nanofiber membrane and non-woven nanofiber membrane.Cell experiments were performed to detect the potential effects of the micropatterned nanofiber membranes on the adhesion,proliferation,migration and extracellular matrix synthesis of fibroblast L929.The effects of lncRNA-XIST overexpression and knockdown on fibroblast L929 cells were observed.The potential effects of miR-150-5p on adhesion,proliferation,migration and extracellular matrix synthesis of fibroblast L929 cells and the underlying mechanism were observed by using miR-150-5p inhibitors and miR-150-5p mimics.Results:In this study,we synthesized nonwoven nanofiber membranes and micropatterned nanofiber membranes using electrospinning,and the nonwoven nanofiber membranes were isotropic with disordered fiber arrangement.However,the micropatterned nanofibrous membranes showed an anisotropic fiber distribution.Both membranes are hydrophobic,and the micropattern structure has no significant effect on the wetting behavior of the nanofiber membrane,but greatly reduces the mechanical properties of the nanofiber membrane.Their effects on wound healing in vivo were tested,and their potential effects on fibroblast L929 cell adhesion,proliferation,migration,and extracellular matrix synthesis were investigated.Our results show that micropatterned nanofiber membranes are more beneficial than non-woven nanofiber membranes in accelerating wound healing,significantly reducing wound area,and significantly promoting epithelial regeneration with more collagen formation and minimal scarring in an in vivo setting.In vitro,the micropatterned nanofiber membrane promoted the adhesion,proliferation,migration and extracellular matrix(α-SMA,COL1A1)synthesis of L929 cells,and stimulated the expression of lncRNA-XIST.In addition,overexpression of lncRNA-XIST using lentivirus significantly inhibited the level of miR-150-5p.In addition,the proliferation and migration of L929 cells and the synthesis of extracellular matrix(α-SMA,COL1A1)were significantly increased.After silencing lncRNA-XIST,the expression of miR-150-5p was significantly up-regulated,while the proliferation and migration of L929 cells and the synthesis of extracellular matrix(α-SMA,COL1A1)were significantly inhibited.miR-150-5p inhibited cell proliferation and migration,and miR-150-5p inhibitor could reverse this inhibitory trend caused by shXIST.Dual-luciferase reporter gene analysis confirmed that miR-150-5p was a putative target gene of lncRNA-XIST,and miR-150-5p competitively bound to COL1A1.Conclusions:Micropatterned nanofiber membrane can promote wound healing by affecting the lncRNA-XIST/miR-150-5p/COL1A1 pathway,which provides a good basis for further understanding of micropatterned nanofiber membrane in promoting wound healing.