北京大学学报(医学版)
北京大學學報(醫學版)
북경대학학보(의학판)
JOURNAL OF BEIJING MEDICAL UNIVERSITY(HEALTH SCIENCES)
2015年
2期
211-218
,共8页
崔云鹏%曹永平%刘恒%杨昕%孟志超%王瑞
崔雲鵬%曹永平%劉恆%楊昕%孟誌超%王瑞
최운붕%조영평%류항%양흔%맹지초%왕서
骨关节炎%间充质干细胞%注射,关节内
骨關節炎%間充質榦細胞%註射,關節內
골관절염%간충질간세포%주사,관절내
Osteoarthritis%Mesenchymal stromal cells%Injection,intra-articular
目的:研究不同浓度骨髓间充质干细胞( bone marrow mesenchymal stem cells,BM-MSCs)对骨关节炎( oste-oarthritis,OA)的治疗效果,并探究其治疗机制。方法:8周龄近交系SD大鼠32只随机分成4组,每组8只,均采用自身双侧对照研究:对照组、高浓度组(1×107/mL BM-MSCs)、低浓度组(5×106/mL BM-MSCs)及高/低浓度对比组。应用改良Hulth方法诱导膝关节OA。对照组一侧行手术,另一侧行假手术,其余各组均行双侧手术。术后4周处死对照组大鼠采集双侧膝关节标本。各试验组大鼠膝关节内注入相应浓度的BM-MSCs或磷酸盐缓冲液( phosphate buffered solution,PBS),切断大鼠双侧坐骨神经及股神经使下肢制动,注射3周后处死各组大鼠并收取双侧膝关节标本。应用Mankin组织学评分评价OA病变程度,RT-PCR检测软骨Ⅱ型胶原mRNA表达,荧光显微镜观察荧光蛋白标记的BM-MSCs在膝关节内分布情况。结果:高浓度组、低浓度组BM-MSCs侧软骨组织标本Mankin评分均明显低于对照侧(5.40±0.51 vs.9.60±0.51;6.60±0.40 vs.10.00±0.32;P均<0.05),高/低浓度对比组高浓度侧Mankin评分略低于低浓度侧(6.40±0.51 vs.7.60±0.75,P>00.5)。 RT-PCR结果显示,高浓度组、低浓度组BM-MSCs侧软骨Ⅱ型胶原mRNA含量分别为对照侧的108%±1%和106%±1%,高/低浓度对比组高浓度侧Ⅱ型胶原mRNA 含量是低浓度侧的102%±1%。荧光显微镜显示,在软骨表面未见绿色荧光蛋白表达,而在滑膜组织内可见绿色荧光表达。结论:关节腔内注入BM-MSCs可能通过间接机制对OA软骨病变起到保护作用,两种浓度治疗效果无差异。
目的:研究不同濃度骨髓間充質榦細胞( bone marrow mesenchymal stem cells,BM-MSCs)對骨關節炎( oste-oarthritis,OA)的治療效果,併探究其治療機製。方法:8週齡近交繫SD大鼠32隻隨機分成4組,每組8隻,均採用自身雙側對照研究:對照組、高濃度組(1×107/mL BM-MSCs)、低濃度組(5×106/mL BM-MSCs)及高/低濃度對比組。應用改良Hulth方法誘導膝關節OA。對照組一側行手術,另一側行假手術,其餘各組均行雙側手術。術後4週處死對照組大鼠採集雙側膝關節標本。各試驗組大鼠膝關節內註入相應濃度的BM-MSCs或燐痠鹽緩遲液( phosphate buffered solution,PBS),切斷大鼠雙側坐骨神經及股神經使下肢製動,註射3週後處死各組大鼠併收取雙側膝關節標本。應用Mankin組織學評分評價OA病變程度,RT-PCR檢測軟骨Ⅱ型膠原mRNA錶達,熒光顯微鏡觀察熒光蛋白標記的BM-MSCs在膝關節內分佈情況。結果:高濃度組、低濃度組BM-MSCs側軟骨組織標本Mankin評分均明顯低于對照側(5.40±0.51 vs.9.60±0.51;6.60±0.40 vs.10.00±0.32;P均<0.05),高/低濃度對比組高濃度側Mankin評分略低于低濃度側(6.40±0.51 vs.7.60±0.75,P>00.5)。 RT-PCR結果顯示,高濃度組、低濃度組BM-MSCs側軟骨Ⅱ型膠原mRNA含量分彆為對照側的108%±1%和106%±1%,高/低濃度對比組高濃度側Ⅱ型膠原mRNA 含量是低濃度側的102%±1%。熒光顯微鏡顯示,在軟骨錶麵未見綠色熒光蛋白錶達,而在滑膜組織內可見綠色熒光錶達。結論:關節腔內註入BM-MSCs可能通過間接機製對OA軟骨病變起到保護作用,兩種濃度治療效果無差異。
목적:연구불동농도골수간충질간세포( bone marrow mesenchymal stem cells,BM-MSCs)대골관절염( oste-oarthritis,OA)적치료효과,병탐구기치료궤제。방법:8주령근교계SD대서32지수궤분성4조,매조8지,균채용자신쌍측대조연구:대조조、고농도조(1×107/mL BM-MSCs)、저농도조(5×106/mL BM-MSCs)급고/저농도대비조。응용개량Hulth방법유도슬관절OA。대조조일측행수술,령일측행가수술,기여각조균행쌍측수술。술후4주처사대조조대서채집쌍측슬관절표본。각시험조대서슬관절내주입상응농도적BM-MSCs혹린산염완충액( phosphate buffered solution,PBS),절단대서쌍측좌골신경급고신경사하지제동,주사3주후처사각조대서병수취쌍측슬관절표본。응용Mankin조직학평분평개OA병변정도,RT-PCR검측연골Ⅱ형효원mRNA표체,형광현미경관찰형광단백표기적BM-MSCs재슬관절내분포정황。결과:고농도조、저농도조BM-MSCs측연골조직표본Mankin평분균명현저우대조측(5.40±0.51 vs.9.60±0.51;6.60±0.40 vs.10.00±0.32;P균<0.05),고/저농도대비조고농도측Mankin평분략저우저농도측(6.40±0.51 vs.7.60±0.75,P>00.5)。 RT-PCR결과현시,고농도조、저농도조BM-MSCs측연골Ⅱ형효원mRNA함량분별위대조측적108%±1%화106%±1%,고/저농도대비조고농도측Ⅱ형효원mRNA 함량시저농도측적102%±1%。형광현미경현시,재연골표면미견록색형광단백표체,이재활막조직내가견록색형광표체。결론:관절강내주입BM-MSCs가능통과간접궤제대OA연골병변기도보호작용,량충농도치료효과무차이。
Objective:To investigate the efficacy of single time intra-articular different concentration of allogeneic bone marrow mesenchymal stem cells ( BM-MSCs ) injection in the treatment of Sprague-Dawley ( SD) rat model of osteoarthritis ( OA) .Methods: In the study, 32 SD rats were equally ran-domized into 4 groups:control group, high concentration group (1 ×107/mL BM-MSCs), low concentra-tion group (5 ×106/mL BM-MSCs) and high vs.low concentration group.The two knees of each rat were set up to a pair.The induction of OA was performed surgically randomly at one side in model group, and bilaterally in the other groups, which were through anterior cruciate ligament transaction ( ACLT) and medial meniscus excising.After the operation, the SD rats were allowed free movement.Four weeks later, different concentrations of allogeneic BM-MSCs isolated from the SD rats, expanded in vitro and suspended in phosphate buffered solution( PBS) were delivered in the articular cavity of both knees;PBS was used as the control.After injection, we excised the femoral nerve and sciatic nerve to disuse the low limb.The cartilage histological sections of knees were scored by Mankin scoring system to assess the se-verity of the pathology.mRNA of collagen Ⅱwas detected by real time polymerase chain reaction ( RT-PCR) .eGFP was detected by fluorescence microscope.Assessments were carried out 4 weeks after the operation in model group, and 3 weeks after injection in the other groups.Results:Mankin scores of the BM-MSCs side and control side were 6.60 ±0.40 vs.10.00 ±0.32 in low concentration group ( P<0.05), and 5.40 ±0.51 vs.9.60 ±0.51 in high concentration group (P<0.05).Mankin scores of high sv.low concentration group were 6.40 ±0.51 vs.7.60 ±0.75 (P>0.05).mRNA expression of collagen Ⅱ of the BM-MSCs side in low concentration group was 106%±1%in contrast to the control side.As in high concentration group it was 108%±1%, and 102%±1%in high vs. low concentra-tion group.Labeled BM-MSCs were detected unexpectedly in the synovial membrane but not in cartilage tissue three weeks from injection.Conclusion:BM-MSCs could promote cartilage repair and inhibit OA progression through a trophic mechanism.There was no difference between the two concentrations.