Peripheral nerve defect repair is a complex process that involves multiple cell types;perineurial cells play a pivotal role.Hair follicle neural crest stem cells promote perineurial cell proliferation and migration vi...Peripheral nerve defect repair is a complex process that involves multiple cell types;perineurial cells play a pivotal role.Hair follicle neural crest stem cells promote perineurial cell proliferation and migration via paracrine signaling;however,their clinical applications are limited by potential risks such as tumorigenesis and xenogeneic immune rejection,which are similar to the risks associated with other stem cell transplantations.The present study therefore focuses on small extracellular vesicles derived from hair follicle neural crest stem cells,which preserve the bioactive properties of the parent cells while avoiding the transplantation-associated risks.In vitro,small extracellular vesicles derived from hair follicle neural crest stem cells significantly enhanced the proliferation,migration,tube formation,and barrier function of perineurial cells,and subsequently upregulated the expression of tight junction proteins.Furthermore,in a rat model of sciatic nerve defects bridged with silicon tubes,treatment with small extracellular vesicles derived from hair follicle neural crest stem cells resulted in higher tight junction protein expression in perineurial cells,thus facilitating neural tissue regeneration.At 10 weeks post-surgery,rats treated with small extracellular vesicles derived from hair follicle neural crest stem cells exhibited improved nerve function recovery and reduced muscle atrophy.Transcriptomic and micro RNA analyses revealed that small extracellular vesicles derived from hair follicle neural crest stem cells deliver mi R-21-5p,which inhibits mothers against decapentaplegic homolog 7 expression,thereby activating the transforming growth factor-β/mothers against decapentaplegic homolog signaling pathway and upregulating hyaluronan synthase 2 expression,and further enhancing tight junction protein expression.Together,our findings indicate that small extracellular vesicles derived from hair follicle neural crest stem cells promote the proliferation,migration,and tight junction protein formation of perineurial cells.These results provide new insights into peripheral nerve regeneration from the perspective of perineurial cells,and present a novel approach for the clinical treatment of peripheral nerve defects.展开更多
目的基于大鼠骨形态发生蛋白2(bone morphogenetic protein2,BMP2)/Smad家族成员1(smad family member 1,Smad1)通路,探究重组人转化生长因子-β1(recombinant human transforming growth factor-β1,rhTGF-β1)对正畸牙移动(orthodonti...目的基于大鼠骨形态发生蛋白2(bone morphogenetic protein2,BMP2)/Smad家族成员1(smad family member 1,Smad1)通路,探究重组人转化生长因子-β1(recombinant human transforming growth factor-β1,rhTGF-β1)对正畸牙移动(orthodontic tooth movement,OMT)大鼠破骨细胞形成的影响。方法构建大鼠OTM模型,采用显微CT(Micro-CT)分析测定OTM的距离;通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色评估压力侧破骨细胞活性;苏木精-伊红(hematoxylin and eosin,HE)染色评估压力侧组织形态学特征,免疫组化(immunohistochemistry,IHC)染色和蛋白质印迹(Western blot)测定相关蛋白表达水平。结果与正常组相比,Model组大鼠OTM距离增加(P<0.01),牙周间隙明显变窄并出现吸收陷窝,压力侧的基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)、核因子κB受体活化因子配体(receptor activator of nuclear factor kappa B ligand,RANKL)增加(P<0.01)、骨保护素(osteoprotegerin,OPG)表达降低(P<0.01),BMP2/Smad1信号通路被激活(P<0.01)。经BMP2抑制剂Noggin处理后,与Model组相比,BMP2、p-Smad1表达显著降低(P<0.01),OTM距离显著降低(P<0.01),且压力侧的TRAP、MMP-9及RANKL表达均显著降低(P<0.01),OPG升高(P<0.01)。经rhTGF-β1处理的大鼠中,较Model组OTM距离显著增加(P<0.01),TRAP阳性多核细胞数量升高(P<0.01),压力侧的MMP-9及RANKL表达均显著升高(P<0.05)、OPG表达显著降低(P<0.01),且BMP2、p-Smad1表达上调(P<0.01)。此外,rhTGF-β1+Noggin组部分逆转了rhTGF-β1组大鼠的破骨细胞数量的增加效应(P<0.01)。结论正畸力可促进破骨细胞形成,且rhTGF-β1可通过BMP2/Smad1信号通路增强OTM过程中破骨细胞的形成。展开更多
基金supported by the National Natural Science Foundation of China,No.81571211(to FL)the Natural Science Foundation of Shanghai,No.22ZR1476800(to CH)。
文摘Peripheral nerve defect repair is a complex process that involves multiple cell types;perineurial cells play a pivotal role.Hair follicle neural crest stem cells promote perineurial cell proliferation and migration via paracrine signaling;however,their clinical applications are limited by potential risks such as tumorigenesis and xenogeneic immune rejection,which are similar to the risks associated with other stem cell transplantations.The present study therefore focuses on small extracellular vesicles derived from hair follicle neural crest stem cells,which preserve the bioactive properties of the parent cells while avoiding the transplantation-associated risks.In vitro,small extracellular vesicles derived from hair follicle neural crest stem cells significantly enhanced the proliferation,migration,tube formation,and barrier function of perineurial cells,and subsequently upregulated the expression of tight junction proteins.Furthermore,in a rat model of sciatic nerve defects bridged with silicon tubes,treatment with small extracellular vesicles derived from hair follicle neural crest stem cells resulted in higher tight junction protein expression in perineurial cells,thus facilitating neural tissue regeneration.At 10 weeks post-surgery,rats treated with small extracellular vesicles derived from hair follicle neural crest stem cells exhibited improved nerve function recovery and reduced muscle atrophy.Transcriptomic and micro RNA analyses revealed that small extracellular vesicles derived from hair follicle neural crest stem cells deliver mi R-21-5p,which inhibits mothers against decapentaplegic homolog 7 expression,thereby activating the transforming growth factor-β/mothers against decapentaplegic homolog signaling pathway and upregulating hyaluronan synthase 2 expression,and further enhancing tight junction protein expression.Together,our findings indicate that small extracellular vesicles derived from hair follicle neural crest stem cells promote the proliferation,migration,and tight junction protein formation of perineurial cells.These results provide new insights into peripheral nerve regeneration from the perspective of perineurial cells,and present a novel approach for the clinical treatment of peripheral nerve defects.
文摘目的基于大鼠骨形态发生蛋白2(bone morphogenetic protein2,BMP2)/Smad家族成员1(smad family member 1,Smad1)通路,探究重组人转化生长因子-β1(recombinant human transforming growth factor-β1,rhTGF-β1)对正畸牙移动(orthodontic tooth movement,OMT)大鼠破骨细胞形成的影响。方法构建大鼠OTM模型,采用显微CT(Micro-CT)分析测定OTM的距离;通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色评估压力侧破骨细胞活性;苏木精-伊红(hematoxylin and eosin,HE)染色评估压力侧组织形态学特征,免疫组化(immunohistochemistry,IHC)染色和蛋白质印迹(Western blot)测定相关蛋白表达水平。结果与正常组相比,Model组大鼠OTM距离增加(P<0.01),牙周间隙明显变窄并出现吸收陷窝,压力侧的基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)、核因子κB受体活化因子配体(receptor activator of nuclear factor kappa B ligand,RANKL)增加(P<0.01)、骨保护素(osteoprotegerin,OPG)表达降低(P<0.01),BMP2/Smad1信号通路被激活(P<0.01)。经BMP2抑制剂Noggin处理后,与Model组相比,BMP2、p-Smad1表达显著降低(P<0.01),OTM距离显著降低(P<0.01),且压力侧的TRAP、MMP-9及RANKL表达均显著降低(P<0.01),OPG升高(P<0.01)。经rhTGF-β1处理的大鼠中,较Model组OTM距离显著增加(P<0.01),TRAP阳性多核细胞数量升高(P<0.01),压力侧的MMP-9及RANKL表达均显著升高(P<0.05)、OPG表达显著降低(P<0.01),且BMP2、p-Smad1表达上调(P<0.01)。此外,rhTGF-β1+Noggin组部分逆转了rhTGF-β1组大鼠的破骨细胞数量的增加效应(P<0.01)。结论正畸力可促进破骨细胞形成,且rhTGF-β1可通过BMP2/Smad1信号通路增强OTM过程中破骨细胞的形成。
