背景:应用固态载体作为细胞支架,修复关节软骨缺损,已有成功经验.尝试将液态载体或凝胶态载体材料复合细胞后注入动物体内,观察该方法的可行性.目的:探讨液态载体或凝胶态载体材料氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2负载同种异体软骨细胞修复全厚关节软骨的可行性.设计:对照实验.单位:威海市立医院骨科,山东省创伤骨科研究所.材料:实验于2001-11/2003-09在山东省创伤骨科研究所实验室进行.健康成年新西兰大白兔36只,体质量2.5～4.5 kg,雌雄不限.编号后根据缺损处注入物质的成分随机数字法分成4组,即氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2软骨细胞组、氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2组、氧化聚乙烯和氧化聚丙烯复合物软骨细胞组、空白对照组,每组9只.方法:36只大白兔分组后造关节软骨缺损模型,取同种新西兰大白兔关节软骨细胞,体外培养扩增后与20％氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2混合,移植到缺损处进行修复.各移植组缺损处分别注入氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2负载软骨细胞、氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2、氧化聚乙烯和氧化聚丙烯复合物软骨细胞组.空白对照组:缺损处不做任何处理.移植后4,8,12周对缺损的修复情况进行大体、光镜组织学评估和电镜观察.据Wakitani评分标准,采用盲法对修复质量作出评价.主要观察指标:①软骨缺损修复程度.②软骨细胞的性质形态,基质中胶原性质、数量及排列方式.结果:①移植的氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2负载软骨细胞中的软骨细胞能很好地生长,4周时,缺损区完全填充.8,12周再生组织与周围正常软骨组织外观相似,界限模糊.组织学检查:形成透明软骨,缺损处被修复.②电镜下:8,12周,修复组织中可见多数成熟的透明软骨细胞及其周围排列不规则的、纤细的、均匀的和无周期性的Ⅱ型胶原.空白对照组仅见纤维修复,再生组织缺乏弹性,表面粗糙,不规则.③修复质量评分:氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2负载软骨细胞组和各组相比在各个时期差异均存在显著性,氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2组和氧化聚乙烯和氧化聚丙烯复合物软骨细胞组与对照组相比在各个时期差异均存在显著性[4周:(3.93±1.91),(4.56±1.07),(4.78±1.09),(8.44±1.13)分；8周:(2.80±1.45),(3.24±1.00),(3.33±1.00),(8.44±1.13)分；12周:(2.22±1.10),(3.01±0.69),(3.00±0.71),(9.00±0.87)分,P＜0.001],但两组之间在各个时期无显著的差异(P＞0.05).结论:氧化聚乙烯和氧化聚丙烯复合物结合重组人骨形态蛋白2负载同种异体软骨细胞移植能以透明软骨的方式成功修复兔膝股骨髁软骨缺损,并优于单纯的氧化聚乙烯和氧化聚丙烯复合物负载重组人骨形态蛋白2或单纯的氧化聚乙烯和氧化聚丙烯复合物软骨细胞的移植.
揹景:應用固態載體作為細胞支架,脩複關節軟骨缺損,已有成功經驗.嘗試將液態載體或凝膠態載體材料複閤細胞後註入動物體內,觀察該方法的可行性.目的:探討液態載體或凝膠態載體材料氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2負載同種異體軟骨細胞脩複全厚關節軟骨的可行性.設計:對照實驗.單位:威海市立醫院骨科,山東省創傷骨科研究所.材料:實驗于2001-11/2003-09在山東省創傷骨科研究所實驗室進行.健康成年新西蘭大白兔36隻,體質量2.5～4.5 kg,雌雄不限.編號後根據缺損處註入物質的成分隨機數字法分成4組,即氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2軟骨細胞組、氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2組、氧化聚乙烯和氧化聚丙烯複閤物軟骨細胞組、空白對照組,每組9隻.方法:36隻大白兔分組後造關節軟骨缺損模型,取同種新西蘭大白兔關節軟骨細胞,體外培養擴增後與20％氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2混閤,移植到缺損處進行脩複.各移植組缺損處分彆註入氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2負載軟骨細胞、氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2、氧化聚乙烯和氧化聚丙烯複閤物軟骨細胞組.空白對照組:缺損處不做任何處理.移植後4,8,12週對缺損的脩複情況進行大體、光鏡組織學評估和電鏡觀察.據Wakitani評分標準,採用盲法對脩複質量作齣評價.主要觀察指標:①軟骨缺損脩複程度.②軟骨細胞的性質形態,基質中膠原性質、數量及排列方式.結果:①移植的氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2負載軟骨細胞中的軟骨細胞能很好地生長,4週時,缺損區完全填充.8,12週再生組織與週圍正常軟骨組織外觀相似,界限模糊.組織學檢查:形成透明軟骨,缺損處被脩複.②電鏡下:8,12週,脩複組織中可見多數成熟的透明軟骨細胞及其週圍排列不規則的、纖細的、均勻的和無週期性的Ⅱ型膠原.空白對照組僅見纖維脩複,再生組織缺乏彈性,錶麵粗糙,不規則.③脩複質量評分:氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2負載軟骨細胞組和各組相比在各箇時期差異均存在顯著性,氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2組和氧化聚乙烯和氧化聚丙烯複閤物軟骨細胞組與對照組相比在各箇時期差異均存在顯著性[4週:(3.93±1.91),(4.56±1.07),(4.78±1.09),(8.44±1.13)分；8週:(2.80±1.45),(3.24±1.00),(3.33±1.00),(8.44±1.13)分；12週:(2.22±1.10),(3.01±0.69),(3.00±0.71),(9.00±0.87)分,P＜0.001],但兩組之間在各箇時期無顯著的差異(P＞0.05).結論:氧化聚乙烯和氧化聚丙烯複閤物結閤重組人骨形態蛋白2負載同種異體軟骨細胞移植能以透明軟骨的方式成功脩複兔膝股骨髁軟骨缺損,併優于單純的氧化聚乙烯和氧化聚丙烯複閤物負載重組人骨形態蛋白2或單純的氧化聚乙烯和氧化聚丙烯複閤物軟骨細胞的移植.
배경:응용고태재체작위세포지가,수복관절연골결손,이유성공경험.상시장액태재체혹응효태재체재료복합세포후주입동물체내,관찰해방법적가행성.목적:탐토액태재체혹응효태재체재료양화취을희화양화취병희복합물결합중조인골형태단백2부재동충이체연골세포수복전후관절연골적가행성.설계:대조실험.단위:위해시립의원골과,산동성창상골과연구소.재료:실험우2001-11/2003-09재산동성창상골과연구소실험실진행.건강성년신서란대백토36지,체질량2.5～4.5 kg,자웅불한.편호후근거결손처주입물질적성분수궤수자법분성4조,즉양화취을희화양화취병희복합물결합중조인골형태단백2연골세포조、양화취을희화양화취병희복합물결합중조인골형태단백2조、양화취을희화양화취병희복합물연골세포조、공백대조조,매조9지.방법:36지대백토분조후조관절연골결손모형,취동충신서란대백토관절연골세포,체외배양확증후여20％양화취을희화양화취병희복합물결합중조인골형태단백2혼합,이식도결손처진행수복.각이식조결손처분별주입양화취을희화양화취병희복합물결합중조인골형태단백2부재연골세포、양화취을희화양화취병희복합물결합중조인골형태단백2、양화취을희화양화취병희복합물연골세포조.공백대조조:결손처불주임하처리.이식후4,8,12주대결손적수복정황진행대체、광경조직학평고화전경관찰.거Wakitani평분표준,채용맹법대수복질량작출평개.주요관찰지표:①연골결손수복정도.②연골세포적성질형태,기질중효원성질、수량급배렬방식.결과:①이식적양화취을희화양화취병희복합물결합중조인골형태단백2부재연골세포중적연골세포능흔호지생장,4주시,결손구완전전충.8,12주재생조직여주위정상연골조직외관상사,계한모호.조직학검사:형성투명연골,결손처피수복.②전경하:8,12주,수복조직중가견다수성숙적투명연골세포급기주위배렬불규칙적、섬세적、균균적화무주기성적Ⅱ형효원.공백대조조부견섬유수복,재생조직결핍탄성,표면조조,불규칙.③수복질량평분:양화취을희화양화취병희복합물결합중조인골형태단백2부재연골세포조화각조상비재각개시기차이균존재현저성,양화취을희화양화취병희복합물결합중조인골형태단백2조화양화취을희화양화취병희복합물연골세포조여대조조상비재각개시기차이균존재현저성[4주:(3.93±1.91),(4.56±1.07),(4.78±1.09),(8.44±1.13)분；8주:(2.80±1.45),(3.24±1.00),(3.33±1.00),(8.44±1.13)분；12주:(2.22±1.10),(3.01±0.69),(3.00±0.71),(9.00±0.87)분,P＜0.001],단량조지간재각개시기무현저적차이(P＞0.05).결론:양화취을희화양화취병희복합물결합중조인골형태단백2부재동충이체연골세포이식능이투명연골적방식성공수복토슬고골과연골결손,병우우단순적양화취을희화양화취병희복합물부재중조인골형태단백2혹단순적양화취을희화양화취병희복합물연골세포적이식.
BACKGROUND: It has been successful to repair articular cartilage defects by using solid carrier as cytoskeleton. We tried to transplant liquid or gel carrier materials combined cells into the body of animals, and investigated its feasibility.OBJECTIVE: To investigate the feasibility of homo-transplatation with liquid or gel carrier materials of Pluronic F-127-recombinant human bone morphogenetic protein-2 (rhBMP-2) engineered chondrocytes for the repair of full-thickness rabbit articular cartilage defect.DESIGN: A controlled experiment.SETTINGS: Department of Orthopaedics, Weihai Municipal Hospital;Shandong Institute of Orthopaedics and Traumaology.MATERIALS: The experiments were carried out in the laboratory of Shandong Institute of Orthopaedics and Traumaology from November 2001 to September 2003. Thirty-six healthy adult New Zealand rabbits of 2.5-4.5 kg, either male or female, were divided into four groups according to the method of random number table: Pluronic F-127-rhBMP-2 engineered chondrocytes group, Pluronic F-127-rhBMP group, Pluronic F127 engineered chondrocytes group and blank control group, with 9 rabbits in each group.METHODS: After grouping, the 36 rabbits were made into models of articular cartilage defects. Pluronic F-127-rhBMP-2 was used as a vector of chondrocytes which were obtained from New Zealand rabbits after cultured and amplified in vitro. The mixture of Pluronic F-127, Pluronic F-127-rhBMP-2 and cultured chondrocytes was transplanted into the defects of articular cartilage that had been made previously with φb3.5 mm drill.There was not any treatment in the blank control group. At 4, 8 and 12 weeks postoperatively, the repairing conditions of the defects were evaluated with gross observation and histological observation under light microscope and under electron microscope. The repaire quality was assessed blindly according to the Wakitani scoring standard.MAIN OUTCOME MEASURES: ① Healing of cartilage defects; ② Property and morphology of the chondrocytes, characteristics, number and arrangement of collagens in matrix.RESULTS: ① In the Pluronic F-127-rhBMP-2 engineered chondrocytes group, the transplanted chondrocytes could grow better than those in other groups, the defected areas were completely filled at 4 weeks. The regenerated tissues at 8 and 12 weeks had similar appearance with the surrounding normal cartilage tissue, but vague. Delimitation. The histological examination showed that transparent cartilages formed, and the defects were healed. ② Under electron microscope at 8 and 12 weeks, there were mature transparent cartilages in the repaired tissues, and there were irregularly arranged slight, even and non-periodical collagen Ⅱ in surrounding. In the blank control group, only fibrous repair was observed, the regenerated tissue lacked elasticity with rough surface. ③ Repairing quality score: The scores at each time point in the Pluronic F-127-rhBMP-2 engineered chondrocytes group were significantly different from those in the other groups.Those in the Pluronic F-127-rhBMP-2 engineered chondrocytes group and Pluronic F-127-rhBMP-2 group and Pluronic F-127 engineered chondrocytes group were significantly different from those in the blank control group [4 weeks: (3.93±1.91), (4.56±1.07), (4.78±1.09), (8.44±1.13) points:8 weeks: (2.80±1.45), (3.24±1.00), (3.33±1.00), (8.44±1.13) points; 12 weeks (2.22±1.10), (3.01±0.69), (3.00±0.71), (9.00±0.87) points, P ＜ 0.001],but there were no significant differences between the two groups (P ＞ 0.05).CONCLUSION: The mixture of Pluronic F-127-rhBMP-2 and cultured chondrocytes can repair successfully the cartilage defects of femoral condyle of rabbit knees by means of hyaline cartilage than simple application of Pluronic F-127-rhBMP-2 or Pluronic F-127 engineered chondrocytes.
