Vascularization and bone regeneration are closely related in the process of bone remodeling,and designing a bioactive scaffold with pro-angiogenic and osteogenic properties may accelerate the repair of bone defects.In...Vascularization and bone regeneration are closely related in the process of bone remodeling,and designing a bioactive scaffold with pro-angiogenic and osteogenic properties may accelerate the repair of bone defects.In this work,an iron-based metal-organic framework(MIL-88)was developed as a carrier for loading a pro-angiogenic small molecular drug(dimethyloxallyl glycine,DMOG),and then embedded into the PLGA nanofibrous scaffolds to repair cranial defects in rats.Imaging and histological evaluation indicated that PLGA/MIL@D scaffold markedly enhanced vascularization and bone regeneration in vivo.Moreover,in vitro assay showed that co-delivery system significantly promoted angiogenesis by stimulating endothelial cell migration,tube formation,and enhanced osteogenesis by promoting expression of osteoblast related proteins.In addition,PLGA/MIL@D scaffold promotes angiogenesis by activating the hypoxia-inducible factor-1(HIF-1)/vascular endothelial growth factor(VEGF)signaling pathway.Altogether,this bioactive PLGA/MIL@D scaffold can combine angiogenesis with osteogenesis,and will be a bright strategy for the repair of bone defects.展开更多
The coupled process of osteogenesis-angiogenesis plays a crucial role in periodontal tissue regeneration.Although various cytokines or chemokines have been widely applied in periodontal in situ tissue engineering,most...The coupled process of osteogenesis-angiogenesis plays a crucial role in periodontal tissue regeneration.Although various cytokines or chemokines have been widely applied in periodontal in situ tissue engineering,most of them are macromolecular proteins with the drawbacks of short effective half-life,poor stability and high cost,which constrain their clinical translation.Our study aimed to develop a difunctional structure for periodontal tissue regeneration by incorporating an angiogenic small molecule,dimethyloxalylglycine(DMOG),and an osteoinductive inorganic nanomaterial,nanosilicate(nSi)into poly(lactic-co-glycolic acid)(PLGA)fibers by electrospinning.The physiochemical properties of DMOG/nSi-PLGA fibrous membranes were characterized.Thereafter,the effect of DMOG/nSi-PLGA membranes on periodontal tissue regeneration was evaluated by detecting osteogenic and angiogenic differentiation potential of periodontal ligament stem cells(PDLSCs)in vitro.Additionally,the fibrous membranes were transplanted into rat periodontal defects,and tissue regeneration was assessed with histological evaluation,micro-computed tomography(micro-CT),and immunohistochemical analysis.DMOG/nSi-PLGA membranes possessed preferable mechanical property and biocompatibility.PDLSCs seeded on the DMOG/nSi-PLGA membranes showed up-regulated expression of osteogenic and angiogenic markers,higher alkaline phosphatase(ALP)activity,and more tube formation in comparison with single application.Further,in vivo study showed that the DMOG/nSi-PLGA membranes promoted recruitment of CD90+/CD34stromal cells,induced angiogenesis and osteogenesis,and regenerated cementum-ligament-bone complex in periodontal defects.Consequently,the combination of DMOG and nSi exerted admirable effects on periodontal tissue regeneration.DMOG/nSi-PLGA fibrous membranes could enhance and orchestrate osteogenesis-angiogenesis,and may have the potential to be translated as an effective scaffold in periodontal tissue engineering.展开更多
文摘目的探讨在缺氧缺血清环境下,脯氨酸羟化酶抑制剂——二甲基乙二酰甘氨酸(DMOG)促进骨髓间充质干细胞(MSCs)血管新生的作用及其机制。方法 MSCs从大鼠骨髓中分离得到,分为:常氧条件下溶剂对照组(N-DMSO)、缺氧条件下溶剂对照组(H-DMSO)、20μM DMOG加药组(D-20μM)、100μM DMOG加药组(D-100μM)及500μM DMOG加药组(D-500μM),观察以下指标:(1)通过缺氧条件下CCK-8检测DMOG在20μM、100μM及500μM剂量下对MSCs的保护作用,确定DMOG的最佳剂量;(2)通过Matrigel实验观察DMOG促进MSCs血管新生的作用;(3)通过Western blot法检测血管新生通路相关蛋白表达。结果 (1)不同浓度组DMOG对缺氧损伤MSCs的影响:N-DMSO组OD值3.25±0.05,H-DMSO组2.23±0.14,D-20μM组2.68±0.43,D-100μM组3.11±0.25,D-500μM组3.23±0.04。与N-DMSO组比较,H-DMSO组OD值降低(P<0.01),与H-DMSO组比较,D-20μM组、D-100μM组、D-500μM OD值均升高(P<0.05或0.01)。(2)不同浓度组DMOG对正常MSCs的影响:D-20μM组3.19±0.02,D-100μM组3.15±0.06,D-500μM组2.51±0.08。与N-DMSO组比较,D-500μM组OD值降低(P<0.01),D-20μM组、D-100μM组与N-DMSO组比较差异均无统计学意义(均P>0.05),提示DMOG在20μM及100μM对细胞无毒性。(3)血管新生相关蛋白的表达:与H-DMSO组相比,D-100μM组缺氧3、6及24h HIF-1α均增加(1.44±0.32 vs 7.79±0.23,2.4±0.28 vs 3.51±0.79,0.93±0.37 vs2.46±0.07,P<0.05或0.01),p AKT则在缺氧6h增加(0.47±0.15 vs 0.71±0.03,P<0.05),VEGF在缺氧6、24h均增加(0.63±0.10 vs 0.87±0.14,0.42±0.06 vs 0.70±0.06,P<0.05或0.01),以缺氧24h最为显著。pm TOR/m TOR及p ERK/ERK蛋白两组比较差异均无统计学意义(均P>0.05)。结论 DMOG通过抑制HIF-1α降解及促进AKT磷酸化提高缺氧环境下MSCs的血管新生能力。
基金supported by the Fundamental Research Funds for the Central Universities of China(No.DUT22YG201)。
文摘Vascularization and bone regeneration are closely related in the process of bone remodeling,and designing a bioactive scaffold with pro-angiogenic and osteogenic properties may accelerate the repair of bone defects.In this work,an iron-based metal-organic framework(MIL-88)was developed as a carrier for loading a pro-angiogenic small molecular drug(dimethyloxallyl glycine,DMOG),and then embedded into the PLGA nanofibrous scaffolds to repair cranial defects in rats.Imaging and histological evaluation indicated that PLGA/MIL@D scaffold markedly enhanced vascularization and bone regeneration in vivo.Moreover,in vitro assay showed that co-delivery system significantly promoted angiogenesis by stimulating endothelial cell migration,tube formation,and enhanced osteogenesis by promoting expression of osteoblast related proteins.In addition,PLGA/MIL@D scaffold promotes angiogenesis by activating the hypoxia-inducible factor-1(HIF-1)/vascular endothelial growth factor(VEGF)signaling pathway.Altogether,this bioactive PLGA/MIL@D scaffold can combine angiogenesis with osteogenesis,and will be a bright strategy for the repair of bone defects.
基金the National Natural Science Foundation of China(No.81670993,81873716,and 81901009)The Construction Engineering Special Fund of“Taishan Scholars”of Shandong Province(No.ts20190975 and tsqn201909180)+1 种基金National Key R&D Program of China(No.2017YFB0405400)Collaborative Innovation Center of Technology and Equipment for Biological Diagnosis and Therapy in Universities of Shandong,The National Key Research and Development Program of China(No.2017YFA0104604),Open Foundation of Shandong Provincial Key Laboratory of Oral Tissue Regeneration(No.SDKQ201901,SDKQ201904).The funders had no role in study design,data collection and analysis,decision to publish or preparation of the manuscript.The authors also thank Prof.Hongyu Zhang and Dr.Yi Wang from Tsinghua University for technical guidance.The authors declare that no financial or other potential competing interests exist with regard to this study.
文摘The coupled process of osteogenesis-angiogenesis plays a crucial role in periodontal tissue regeneration.Although various cytokines or chemokines have been widely applied in periodontal in situ tissue engineering,most of them are macromolecular proteins with the drawbacks of short effective half-life,poor stability and high cost,which constrain their clinical translation.Our study aimed to develop a difunctional structure for periodontal tissue regeneration by incorporating an angiogenic small molecule,dimethyloxalylglycine(DMOG),and an osteoinductive inorganic nanomaterial,nanosilicate(nSi)into poly(lactic-co-glycolic acid)(PLGA)fibers by electrospinning.The physiochemical properties of DMOG/nSi-PLGA fibrous membranes were characterized.Thereafter,the effect of DMOG/nSi-PLGA membranes on periodontal tissue regeneration was evaluated by detecting osteogenic and angiogenic differentiation potential of periodontal ligament stem cells(PDLSCs)in vitro.Additionally,the fibrous membranes were transplanted into rat periodontal defects,and tissue regeneration was assessed with histological evaluation,micro-computed tomography(micro-CT),and immunohistochemical analysis.DMOG/nSi-PLGA membranes possessed preferable mechanical property and biocompatibility.PDLSCs seeded on the DMOG/nSi-PLGA membranes showed up-regulated expression of osteogenic and angiogenic markers,higher alkaline phosphatase(ALP)activity,and more tube formation in comparison with single application.Further,in vivo study showed that the DMOG/nSi-PLGA membranes promoted recruitment of CD90+/CD34stromal cells,induced angiogenesis and osteogenesis,and regenerated cementum-ligament-bone complex in periodontal defects.Consequently,the combination of DMOG and nSi exerted admirable effects on periodontal tissue regeneration.DMOG/nSi-PLGA fibrous membranes could enhance and orchestrate osteogenesis-angiogenesis,and may have the potential to be translated as an effective scaffold in periodontal tissue engineering.
