Gene therapy constitutes a promising strategy for the treatment of osteoarthritis (OA). We assessed the use of electroporation (EP) of non-viral gene vectors, and compared its efficacy with that of adeno-associated vi...Gene therapy constitutes a promising strategy for the treatment of osteoarthritis (OA). We assessed the use of electroporation (EP) of non-viral gene vectors, and compared its efficacy with that of adeno-associated virus (AAV) vectors. EP- and AAV-mediated delivery of human interleukin-1 receptor antagonist (hIL-1Ra) was localized performed in the joints of rats following induction of OA. mRNA levels for hIL-1Ra, IL-1β, TNF-α, MMP-13 and ADAMTS-4 in the cartilage and synovial tissues were analyzed. Structural analyses of the subchondral bone at the medial femoral condyle were performed by Micro-CT after treatment. Knee joint specimens were staining with hematoxylin and eosin and Saffron O. Induction of hIL-1Ra by both EP and AAV inhibited inflammatory-induced sub-chondral bone reconstruction, and effectively suppressed IL-1β activity, as evidenced by decreased expression of MMP-13 and ADAMTS-4. Histological analyses revealed significant protection of cartilage, proteoglycan by EP and AAV. hIL-1Ra expression was similar in both the EP and AAV groups. Notably, this gene is not easier degraded transduced by EP compared with AAV. Taken together, these results show that EP offers transfection efficiency comparable to that of AAV, with the potential for longer gene expression, making EP a promising candidate for efficient non-viral delivery of OA gene therapy.展开更多
Mesenchymal stem cells(MSCs)therapy shows the potential benefits to relieve clinical symptoms of osteoarthritis(OA),but it is uncertain if it can repair articular cartilage lesions-the main pathology of OA.Here,we pre...Mesenchymal stem cells(MSCs)therapy shows the potential benefits to relieve clinical symptoms of osteoarthritis(OA),but it is uncertain if it can repair articular cartilage lesions-the main pathology of OA.Here,we prepared biomimetic cupper sulfide@phosphatidylcholine(CuS@PC)nanoparticles(NPs)loaded with plasmid DNA(pDNA)encoding transforming growth factor-beta 1(TGF-β1)to engineer MSCs for enhanced OA therapy via cartilage regeneration.We found that the NPs not only promoted cell proliferation and migration,but also presented a higher pDNA transfection efficiency relative to commercial transfection reagent lipofectamine 3000.The resultant CuS/TGF-β1@PC NP-engineered MSCs(termed CTP-MSCs)were better than pure MSCs in terms of chondrogenic gene expression,glycosaminoglycan deposition and type II collagen formation,favoring cartilage repair.Further,CTP-MSCs inhibited extracellular matrix degradation in interleukin-1β-induced chondrocytes.Consequently,intraarticular administration of CTP-MSCs significantly enhanced the repair of damaged cartilage,whereas pure MSCs exhibited very limited effects on cartilage regeneration in destabilization of the medial meniscus(DMM)surgical instability mice.Hence,this work provides a new strategy to overcome the limitation of current stem cell therapy in OA treatment through developing more effective nanoengineered MSCs.展开更多
背景:在骨关节炎的软骨退变过程中,以软骨细胞为靶点的细胞因子和信号通路起到了重要作用。目的:综述近年来与骨关节炎相关的细胞因子和信号通路的作用机制及治疗方式等最新研究进展,以期为将来探索新的治疗靶点和治疗方式作参考依据。...背景:在骨关节炎的软骨退变过程中,以软骨细胞为靶点的细胞因子和信号通路起到了重要作用。目的:综述近年来与骨关节炎相关的细胞因子和信号通路的作用机制及治疗方式等最新研究进展,以期为将来探索新的治疗靶点和治疗方式作参考依据。方法:在中国知网、万方数据、维普、PubMed、Web of Science和MEDLINE数据库进行文献检索,并以“骨关节炎,细胞因子,信号通路,软骨细胞,炎症,治疗”作为中文检索词,以“osteoarthritis,cytokine,signal pathway,chondrocyte,inflammation,therapy”作为英文检索词,对最终纳入的60篇文献进行了归纳总结。结果与结论:(1)在目前的研究中认为骨关节炎具体机制尚不明确,大量研究表明骨关节炎与细胞因子和信号通路关系密切,是一个复杂的作用过程,以细胞因子和信号通路作为治疗突破口的相关研究也是当前的热点。(2)白细胞介素1等促炎因子的受体拮抗剂对于骨关节炎治疗效果不佳,已有更多的学者开始进行基因治疗方式的研究。(3)目前关于转化生长因子β等细胞因子和Wnt信号通路的重组因子、基因治疗和间充质干细胞等治疗方式均获得了积极的研究成果,但其安全性和有效性等问题还需进一步的基础和临床研究验证。(4)目前相关的骨关节炎治疗方式如富血小板血浆疗法已广泛应用于临床,而重组因子疗法、基因治疗和间充质干细胞治疗均处于研究阶段,其中间充质干细胞治疗和基因治疗有望在软骨修复和再生领域做出突破,是值得期待的治疗方式,但其有效性和安全性尚需进一步研究,未来需探寻其作用机制和适用范围以制定标准。展开更多
Knee osteoarthritis is the most common joint disease.It causes pain and suffering for affected patients and is the source of major economic costs for healthcare systems.Despite ongoing research,there is a lack of know...Knee osteoarthritis is the most common joint disease.It causes pain and suffering for affected patients and is the source of major economic costs for healthcare systems.Despite ongoing research,there is a lack of knowledge regarding disease mechanisms,biomarkers,and possible cures.Current treatments do not fulfill patients’long-term needs,and it often requires invasive surgical procedures with subsequent long periods of rehabilitation.Researchers and companies worldwide are working to find a suitable cell source to engineer or regenerate a functional and healthy articular cartilage tissue to implant in the damaged area.Potential cell sources to accomplish this goal include embryonic stem cells,mesenchymal stem cells,or induced pluripotent stem cells.The differentiation of stem cells into different tissue types is complex,and a suitable concentration range of specific growth factors is vital.The cellular microenvironment during early embryonic development provides crucial information regarding concentrations of signaling molecules and morphogen gradients as these are essential inducers for tissue development.Thus,morphogen gradients implemented in developmental protocols aimed to engineer functional cartilage tissue can potentially generate cells comparable to those within native cartilage.In this review,we have summarized the problems with current treatments,potential cell sources for cell therapy,reviewed the progress of new treatments within the regenerative cartilage field,and highlighted the importance of cell quality,characterization assays,and chemically defined protocols.展开更多
文摘Gene therapy constitutes a promising strategy for the treatment of osteoarthritis (OA). We assessed the use of electroporation (EP) of non-viral gene vectors, and compared its efficacy with that of adeno-associated virus (AAV) vectors. EP- and AAV-mediated delivery of human interleukin-1 receptor antagonist (hIL-1Ra) was localized performed in the joints of rats following induction of OA. mRNA levels for hIL-1Ra, IL-1β, TNF-α, MMP-13 and ADAMTS-4 in the cartilage and synovial tissues were analyzed. Structural analyses of the subchondral bone at the medial femoral condyle were performed by Micro-CT after treatment. Knee joint specimens were staining with hematoxylin and eosin and Saffron O. Induction of hIL-1Ra by both EP and AAV inhibited inflammatory-induced sub-chondral bone reconstruction, and effectively suppressed IL-1β activity, as evidenced by decreased expression of MMP-13 and ADAMTS-4. Histological analyses revealed significant protection of cartilage, proteoglycan by EP and AAV. hIL-1Ra expression was similar in both the EP and AAV groups. Notably, this gene is not easier degraded transduced by EP compared with AAV. Taken together, these results show that EP offers transfection efficiency comparable to that of AAV, with the potential for longer gene expression, making EP a promising candidate for efficient non-viral delivery of OA gene therapy.
基金supported by National Natural Science Foundation of China(81902198,81974342,82172391)Guangdong Basic and Applied Basic Research Foundation(2020A1515010398)+2 种基金China National Postdoctoral Program for Innovative Talents(BX20190150)China Postdoctoral Science Foundation(2019M662980)President Foundation of Zhujiang Hospital,Southern Medical University(yzjj2018rc09).
文摘Mesenchymal stem cells(MSCs)therapy shows the potential benefits to relieve clinical symptoms of osteoarthritis(OA),but it is uncertain if it can repair articular cartilage lesions-the main pathology of OA.Here,we prepared biomimetic cupper sulfide@phosphatidylcholine(CuS@PC)nanoparticles(NPs)loaded with plasmid DNA(pDNA)encoding transforming growth factor-beta 1(TGF-β1)to engineer MSCs for enhanced OA therapy via cartilage regeneration.We found that the NPs not only promoted cell proliferation and migration,but also presented a higher pDNA transfection efficiency relative to commercial transfection reagent lipofectamine 3000.The resultant CuS/TGF-β1@PC NP-engineered MSCs(termed CTP-MSCs)were better than pure MSCs in terms of chondrogenic gene expression,glycosaminoglycan deposition and type II collagen formation,favoring cartilage repair.Further,CTP-MSCs inhibited extracellular matrix degradation in interleukin-1β-induced chondrocytes.Consequently,intraarticular administration of CTP-MSCs significantly enhanced the repair of damaged cartilage,whereas pure MSCs exhibited very limited effects on cartilage regeneration in destabilization of the medial meniscus(DMM)surgical instability mice.Hence,this work provides a new strategy to overcome the limitation of current stem cell therapy in OA treatment through developing more effective nanoengineered MSCs.
文摘背景:在骨关节炎的软骨退变过程中,以软骨细胞为靶点的细胞因子和信号通路起到了重要作用。目的:综述近年来与骨关节炎相关的细胞因子和信号通路的作用机制及治疗方式等最新研究进展,以期为将来探索新的治疗靶点和治疗方式作参考依据。方法:在中国知网、万方数据、维普、PubMed、Web of Science和MEDLINE数据库进行文献检索,并以“骨关节炎,细胞因子,信号通路,软骨细胞,炎症,治疗”作为中文检索词,以“osteoarthritis,cytokine,signal pathway,chondrocyte,inflammation,therapy”作为英文检索词,对最终纳入的60篇文献进行了归纳总结。结果与结论:(1)在目前的研究中认为骨关节炎具体机制尚不明确,大量研究表明骨关节炎与细胞因子和信号通路关系密切,是一个复杂的作用过程,以细胞因子和信号通路作为治疗突破口的相关研究也是当前的热点。(2)白细胞介素1等促炎因子的受体拮抗剂对于骨关节炎治疗效果不佳,已有更多的学者开始进行基因治疗方式的研究。(3)目前关于转化生长因子β等细胞因子和Wnt信号通路的重组因子、基因治疗和间充质干细胞等治疗方式均获得了积极的研究成果,但其安全性和有效性等问题还需进一步的基础和临床研究验证。(4)目前相关的骨关节炎治疗方式如富血小板血浆疗法已广泛应用于临床,而重组因子疗法、基因治疗和间充质干细胞治疗均处于研究阶段,其中间充质干细胞治疗和基因治疗有望在软骨修复和再生领域做出突破,是值得期待的治疗方式,但其有效性和安全性尚需进一步研究,未来需探寻其作用机制和适用范围以制定标准。
基金Most funding by Cline Scientific AB and partly funded by the European Union’s Horizon 2020 research and innovation program under grant agreement RESTORE no 814558 and AUTOCRAT no 874671.
文摘Knee osteoarthritis is the most common joint disease.It causes pain and suffering for affected patients and is the source of major economic costs for healthcare systems.Despite ongoing research,there is a lack of knowledge regarding disease mechanisms,biomarkers,and possible cures.Current treatments do not fulfill patients’long-term needs,and it often requires invasive surgical procedures with subsequent long periods of rehabilitation.Researchers and companies worldwide are working to find a suitable cell source to engineer or regenerate a functional and healthy articular cartilage tissue to implant in the damaged area.Potential cell sources to accomplish this goal include embryonic stem cells,mesenchymal stem cells,or induced pluripotent stem cells.The differentiation of stem cells into different tissue types is complex,and a suitable concentration range of specific growth factors is vital.The cellular microenvironment during early embryonic development provides crucial information regarding concentrations of signaling molecules and morphogen gradients as these are essential inducers for tissue development.Thus,morphogen gradients implemented in developmental protocols aimed to engineer functional cartilage tissue can potentially generate cells comparable to those within native cartilage.In this review,we have summarized the problems with current treatments,potential cell sources for cell therapy,reviewed the progress of new treatments within the regenerative cartilage field,and highlighted the importance of cell quality,characterization assays,and chemically defined protocols.
