CAJ | 학술논문

目的探讨透明质酸定向诱导骨髓MSCs分化修复兔OA模型软骨缺损的作用.方法分离、纯化并扩增兔骨髓MSCs,5-溴-2'脱氧脲嘧啶核苷(BrdU)和4 ’6二脒基-2-苯吲哚盐酸(DAPI)标记.Muehlema法构建兔OA软骨缺损模型24只,并按照随机数字表法随机分为3组:透明质酸治疗组(Th组)、MSCs治疗组(Tm组)、透明质酸联合MSCs治疗组(Th+m组).治疗侧膝关节分别予以相应治疗,对照侧膝关节以等量0.9％氯化钠注射液治疗,每周1次.治疗结束3周后,行单光子发射计算机断层扫描(SPECT)和正电子发射断层扫描/计算机断层扫描(PET-CT)成像.修复标本行大体形态学、组织病理学、免疫组织化学和免疫荧光等检测.采用t检验、方差分析、Newman-Student-Keul检验进行统计分析.结果 ①各组组织病理学Mankin评分治疗侧均低于对照侧(Th组:3.3±0.4,5.3±0.5;Tm组:3.9±0.6,5.4±0.6;Th+m组:2.2±0.3,5.3±0.7),差异均有统计学意义(t=-4.83,-3.83,-13.50;P均＜0.05);Th +m组治疗侧Mankin评分(2.2±0.3)显著低于Th组(3.3±0.4)和Tm组(3.9±0.6),差异有统计学意义(P＜0.01);②PET-CT检测18F标记的2-氟-2-脱氧-D葡萄糖(18F-FDG)放射性核素平均计数率,Th+m治疗组治疗侧(43 773±1 371)显著高于Th组(37 983±1 270)和Tm组(37 097±1 214),差异均有统计学意义(jp＜0.01);③SPECT检测各治疗组治疗侧99Tc标记的亚甲基二膦酸盐(99Tcm-MDP)的放射性核素平均计数率,Th+m治疗组治疗侧(768±19)显著高于Th组(590±16)和Tm组(597±14),差异均有统计学意义(P＜0.01).结论透明质酸联合MSCs移植可以有效地修复软骨缺损.
목적 탐토투명질산정향유도골수MSCs분화수복토OA모형연골결손적작용.방법 분리、순화병확증토골수MSCs,5-추-2'탈양뇨밀정핵감(BrdU)화4 ’6이미기-2-분신타염산(DAPI)표기.Muehlema법구건토OA연골결손모형24지,병안조수궤수자표법수궤분위3조:투명질산치료조(Th조)、MSCs치료조(Tm조)、투명질산연합MSCs치료조(Th+m조).치료측슬관절분별여이상응치료,대조측슬관절이등량0.9％록화납주사액치료,매주1차.치료결속3주후,행단광자발사계산궤단층소묘(SPECT)화정전자발사단층소묘/계산궤단층소묘(PET-CT)성상.수복표본행대체형태학、조직병이학、면역조직화학화면역형광등검측.채용t검험、방차분석、Newman-Student-Keul검험진행통계분석.결과 ①각조조직병이학Mankin평분치료측균저우대조측(Th조:3.3±0.4,5.3±0.5;Tm조:3.9±0.6,5.4±0.6;Th+m조:2.2±0.3,5.3±0.7),차이균유통계학의의(t=-4.83,-3.83,-13.50;P균＜0.05);Th +m조치료측Mankin평분(2.2±0.3)현저저우Th조(3.3±0.4)화Tm조(3.9±0.6),차이유통계학의의(P＜0.01);②PET-CT검측18F표기적2-불-2-탈양-D포도당(18F-FDG)방사성핵소평균계수솔,Th+m치료조치료측(43 773±1 371)현저고우Th조(37 983±1 270)화Tm조(37 097±1 214),차이균유통계학의의(jp＜0.01);③SPECT검측각치료조치료측99Tc표기적아갑기이련산염(99Tcm-MDP)적방사성핵소평균계수솔,Th+m치료조치료측(768±19)현저고우Th조(590±16)화Tm조(597±14),차이균유통계학의의(P＜0.01).결론 투명질산연합MSCs이식가이유효지수복연골결손.
Objective To analyze the effect of bone marrow (BM) derived mesenchymal stem cells (MSCs) induced by hyaluronic acid (HA) on the repair of cartilage defect of rabbit osteoarthritis (OA) model.Methods BM derived MSCs of rabbits were isolated,purified and expanded in vitro,labeled with 5-bromo2-deoxyuridine (BrdU) and 4'6-diamidino-2-phenylindole dihydrochloride (DAPI).Twenty-four rabbits were used to establish OA model of cartilage defect by Muehlema method,and then according to random number table randomly divided into 3 groups including HA treatment group (Th),MSCs treatment group (Tm),and the MSCs induced by HA treatment group (Th+m).One knee of the model rabbit was treated with the corresponding intra-articular injection once a week,while the other knee was injected with the same amount of saline as control Three weeks after treatment,rabbit models were scanned with single-photon emission computed tomography (SPECT) and positron emission tomography/computed tomography (PET-CT).Knee repair specimens were evaluated by gross morphology,histopathology,immunohistochemical and fluorescence staining.All data were analyzed by t test,one-way ANOVA test and Newman-Student-Keul test.Results ① The histopathological scores of the treated knees of the treated knees of the 3 treatment groups were significantly lower than those of the control knees (Th group:3.3±0.4,5.3±0.5; Tm group:3.9±0.6,5.4±0.6; Th+m group:2.2±0.3,5.3±0.7)(t=-4.83,-3.83,-13.50; P＜0.05).The histopathological scores of the treated knees of the Th+m group (2.2±0.3) were significantly lower than those of the Th group (3.3±0.4) and Tm group (3.9±0.6) (P＜O.01).② PET-CT scan assay showed that the average of 18F-fluorodeoxyglucose (FDG) radionuclidecount rate of the treated knees of the Th+m group (43 773±1 371) was significantly higher than that of the Th group (37 983±1 270) and Tm group (37 097±1 214) (P＜0.01).③ SPECT scan assay showed that the 99Tcm-methylenediphosphonate (99Tcm-MDP) radionuclide average count of the treated knees of the Th+m group(768±19) was significantly higher than that of the Th group (590±16) and Tm group (597± 14) (P＜0.01).Conclusion Intra-articular injection of HA induced BM derived MSCs can effectively repair of cartilage defects of OA.