BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene ma...BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.展开更多
采用单一变量法研究了激光功率和温度对激光沉积制造Ti65钛合金高周疲劳性能的影响,通过光学显微镜、超声波探测仪和扫描电镜(scanning electron microscope,SEM)对试样的显微组织、内部缺陷和断口形貌进行了分析.结果表明,不同功率试...采用单一变量法研究了激光功率和温度对激光沉积制造Ti65钛合金高周疲劳性能的影响,通过光学显微镜、超声波探测仪和扫描电镜(scanning electron microscope,SEM)对试样的显微组织、内部缺陷和断口形貌进行了分析.结果表明,不同功率试样的显微组织均为网篮组织,α相的含量明显高于β相的含量,高温下网篮组织中的α相发生粗化,晶粒内部出现一些块状α相,组织的均匀性下降.室温和高温条件下高功率和低功率试样的疲劳极限分别为454,398.5,371.5 MPa和336.25 MPa,同一温度下,高功率试样的疲劳极限要比低功率试样的疲劳极限高10%以上;同一功率下,室温试样的疲劳极限要比高温试样的疲劳极限高18%以上,温度对于高周疲劳的影响更大.激光沉积制造Ti65钛合金试样内部存在气孔缺陷,低功率试样中气孔的数目多且直径大,其疲劳源均形核于气孔缺陷处,气孔直径越大,距离表面距离越近,裂纹萌生的越快,疲劳寿命越低,高功率试样中气孔的数目少且直径小,其疲劳源均萌生于表面裂纹,缺陷的存在对裂纹的萌生存在很大影响.展开更多
文摘BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.
文摘采用单一变量法研究了激光功率和温度对激光沉积制造Ti65钛合金高周疲劳性能的影响,通过光学显微镜、超声波探测仪和扫描电镜(scanning electron microscope,SEM)对试样的显微组织、内部缺陷和断口形貌进行了分析.结果表明,不同功率试样的显微组织均为网篮组织,α相的含量明显高于β相的含量,高温下网篮组织中的α相发生粗化,晶粒内部出现一些块状α相,组织的均匀性下降.室温和高温条件下高功率和低功率试样的疲劳极限分别为454,398.5,371.5 MPa和336.25 MPa,同一温度下,高功率试样的疲劳极限要比低功率试样的疲劳极限高10%以上;同一功率下,室温试样的疲劳极限要比高温试样的疲劳极限高18%以上,温度对于高周疲劳的影响更大.激光沉积制造Ti65钛合金试样内部存在气孔缺陷,低功率试样中气孔的数目多且直径大,其疲劳源均形核于气孔缺陷处,气孔直径越大,距离表面距离越近,裂纹萌生的越快,疲劳寿命越低,高功率试样中气孔的数目少且直径小,其疲劳源均萌生于表面裂纹,缺陷的存在对裂纹的萌生存在很大影响.
