中国骨与关节杂志
中國骨與關節雜誌
중국골여관절잡지
Chinese Journal of Bone and Joint
2015年
4期
280-287
,共8页
王硕%谢惠敏%甘少磊%任卫卫%陈秉耀%张增亮%韦兴
王碩%謝惠敏%甘少磊%任衛衛%陳秉耀%張增亮%韋興
왕석%사혜민%감소뢰%임위위%진병요%장증량%위흥
BMPs%活性多肽%骨修复材料%异位成骨
BMPs%活性多肽%骨脩複材料%異位成骨
BMPs%활성다태%골수복재료%이위성골
Bone morphogenetic protein ( BMP )%Active polypeptide%Bone repair material%Ectopic bone formation
目的：通过检测不同浓度骨形态发生蛋白( bone morphogenetic proteins，BMP ) BMPIII 与 BMPIV 活性多肽在大鼠体内异位成骨情况，来评价两种多肽在动物体内骨诱导能力；并与已检测 BMPI 0.4 g / L 浓度进行对比，以选出最优多肽组及浓度。方法应用随机抽签法，将48只 SD 大鼠随机分为8组( A、B、C、D、E、F、G、H 组)，每组6只。将2种多肽的三种不同浓度(0.2 g / L、0.4 g / L、0.8 g / L 的 BMPIII 和0.2 g / L、0.4 g / L、0.8 g / L 的 BMPIV )分别与羟基磷灰石聚乳酸( hydroxyapatite and poly lactic acid， HA / PLA )组成复合材料植入大鼠臀部肌肉浅层，A、B、C、D、E、F 组；将0.4 g / L 浓度 BMPI 多肽与 HA / PLA 组成的复合材料植入的实验对照为 G 组，仅植入 HA / PLA 框架材料空白对照为 H 组。术后3周摄背部正位 X 线片；术后5周时进行 X 线、CT 照射。并于术后3周、5周分别取出标本，采用 HE、Masson 染色在光镜下观察，应用 Masson 染色评分并进行非参数统计分析大鼠术后5周的成骨特性。结果所有动物均顺利完成手术，术后观察期间，动物饮食、活动良好，伤口愈合良好，全部存活。植入材料各组硬度均增加。(1) X 线检查：术后3周，手术植入多肽材料部位均有轻度模糊显影区，其中 C、F 组显影明显；术后5周，A、D 组植入区内有较浅显影，B、E、G 组有较浅较大显影，C、F 组显影较明显；H 组未见明显显影。(2) CT 显示：除 A、D 组可见较模糊低密度显影外，B、C、E、F、G 组均显影明显，H 组未见显影。(3) HE 染色：植入多肽材料的各组术后3周可见少量成骨细胞长入多孔材料，贴附于孔壁；术后5周材料部分降解，其中 B、C、E、F、G 组有较多软骨基质和软骨细胞形成，H 组框架材料分解极少。(4) Masson 染色：术后3周，A、D 组在降解材料周围仅有少量软骨胶原形成，且 B、C 组和 E、F 组软骨胶原量分别明显多于 A 组和 D 组，G 组蓝色面积较小，H 组降解材料周围软骨胶原极少；术后5周，植入多肽材料的各组蓝色面积明显多于术后3周，H 组降解材料周围软骨胶原较少，仅见少量蓝色软骨胶原形成。(5) Masson 评分：B、C、E、F 组分别为(2.22±0.45)分、(2.44±0.3)分、(2.28±0.25)分、(2.44±0.35)分均明显大于 A 组(1.06±0.39)分、D 组(0.72±0.25)分，差异有统计学意义(P＜0.05)；B、C、E、F 组各组间评分两两比较，差异均无统计学意义(P＞0.05)；G 组评分(2.67±0.30)分较 B、E 组大，差异均有统计学意义(P＜0.05)；H 组评分最少，为(0.222±0.27)分。结论 BMPIII 与 BMPIV 两种活性多肽人工复合材料均具有骨诱导能力；0.4 g / L 和0.8 g / L 浓度骨诱导能力较0.2 g / L 浓度大；0.4 g / L BMPI 骨诱导能力较 BMPIII 与 BMPIV 大，更适于成为骨诱导材料。
目的：通過檢測不同濃度骨形態髮生蛋白( bone morphogenetic proteins，BMP ) BMPIII 與 BMPIV 活性多肽在大鼠體內異位成骨情況，來評價兩種多肽在動物體內骨誘導能力；併與已檢測 BMPI 0.4 g / L 濃度進行對比，以選齣最優多肽組及濃度。方法應用隨機抽籤法，將48隻 SD 大鼠隨機分為8組( A、B、C、D、E、F、G、H 組)，每組6隻。將2種多肽的三種不同濃度(0.2 g / L、0.4 g / L、0.8 g / L 的 BMPIII 和0.2 g / L、0.4 g / L、0.8 g / L 的 BMPIV )分彆與羥基燐灰石聚乳痠( hydroxyapatite and poly lactic acid， HA / PLA )組成複閤材料植入大鼠臀部肌肉淺層，A、B、C、D、E、F 組；將0.4 g / L 濃度 BMPI 多肽與 HA / PLA 組成的複閤材料植入的實驗對照為 G 組，僅植入 HA / PLA 框架材料空白對照為 H 組。術後3週攝揹部正位 X 線片；術後5週時進行 X 線、CT 照射。併于術後3週、5週分彆取齣標本，採用 HE、Masson 染色在光鏡下觀察，應用 Masson 染色評分併進行非參數統計分析大鼠術後5週的成骨特性。結果所有動物均順利完成手術，術後觀察期間，動物飲食、活動良好，傷口愈閤良好，全部存活。植入材料各組硬度均增加。(1) X 線檢查：術後3週，手術植入多肽材料部位均有輕度模糊顯影區，其中 C、F 組顯影明顯；術後5週，A、D 組植入區內有較淺顯影，B、E、G 組有較淺較大顯影，C、F 組顯影較明顯；H 組未見明顯顯影。(2) CT 顯示：除 A、D 組可見較模糊低密度顯影外，B、C、E、F、G 組均顯影明顯，H 組未見顯影。(3) HE 染色：植入多肽材料的各組術後3週可見少量成骨細胞長入多孔材料，貼附于孔壁；術後5週材料部分降解，其中 B、C、E、F、G 組有較多軟骨基質和軟骨細胞形成，H 組框架材料分解極少。(4) Masson 染色：術後3週，A、D 組在降解材料週圍僅有少量軟骨膠原形成，且 B、C 組和 E、F 組軟骨膠原量分彆明顯多于 A 組和 D 組，G 組藍色麵積較小，H 組降解材料週圍軟骨膠原極少；術後5週，植入多肽材料的各組藍色麵積明顯多于術後3週，H 組降解材料週圍軟骨膠原較少，僅見少量藍色軟骨膠原形成。(5) Masson 評分：B、C、E、F 組分彆為(2.22±0.45)分、(2.44±0.3)分、(2.28±0.25)分、(2.44±0.35)分均明顯大于 A 組(1.06±0.39)分、D 組(0.72±0.25)分，差異有統計學意義(P＜0.05)；B、C、E、F 組各組間評分兩兩比較，差異均無統計學意義(P＞0.05)；G 組評分(2.67±0.30)分較 B、E 組大，差異均有統計學意義(P＜0.05)；H 組評分最少，為(0.222±0.27)分。結論 BMPIII 與 BMPIV 兩種活性多肽人工複閤材料均具有骨誘導能力；0.4 g / L 和0.8 g / L 濃度骨誘導能力較0.2 g / L 濃度大；0.4 g / L BMPI 骨誘導能力較 BMPIII 與 BMPIV 大，更適于成為骨誘導材料。
목적：통과검측불동농도골형태발생단백( bone morphogenetic proteins，BMP ) BMPIII 여 BMPIV 활성다태재대서체내이위성골정황，래평개량충다태재동물체내골유도능력；병여이검측 BMPI 0.4 g / L 농도진행대비，이선출최우다태조급농도。방법응용수궤추첨법，장48지 SD 대서수궤분위8조( A、B、C、D、E、F、G、H 조)，매조6지。장2충다태적삼충불동농도(0.2 g / L、0.4 g / L、0.8 g / L 적 BMPIII 화0.2 g / L、0.4 g / L、0.8 g / L 적 BMPIV )분별여간기린회석취유산( hydroxyapatite and poly lactic acid， HA / PLA )조성복합재료식입대서둔부기육천층，A、B、C、D、E、F 조；장0.4 g / L 농도 BMPI 다태여 HA / PLA 조성적복합재료식입적실험대조위 G 조，부식입 HA / PLA 광가재료공백대조위 H 조。술후3주섭배부정위 X 선편；술후5주시진행 X 선、CT 조사。병우술후3주、5주분별취출표본，채용 HE、Masson 염색재광경하관찰，응용 Masson 염색평분병진행비삼수통계분석대서술후5주적성골특성。결과소유동물균순리완성수술，술후관찰기간，동물음식、활동량호，상구유합량호，전부존활。식입재료각조경도균증가。(1) X 선검사：술후3주，수술식입다태재료부위균유경도모호현영구，기중 C、F 조현영명현；술후5주，A、D 조식입구내유교천현영，B、E、G 조유교천교대현영，C、F 조현영교명현；H 조미견명현현영。(2) CT 현시：제 A、D 조가견교모호저밀도현영외，B、C、E、F、G 조균현영명현，H 조미견현영。(3) HE 염색：식입다태재료적각조술후3주가견소량성골세포장입다공재료，첩부우공벽；술후5주재료부분강해，기중 B、C、E、F、G 조유교다연골기질화연골세포형성，H 조광가재료분해겁소。(4) Masson 염색：술후3주，A、D 조재강해재료주위부유소량연골효원형성，차 B、C 조화 E、F 조연골효원량분별명현다우 A 조화 D 조，G 조람색면적교소，H 조강해재료주위연골효원겁소；술후5주，식입다태재료적각조람색면적명현다우술후3주，H 조강해재료주위연골효원교소，부견소량람색연골효원형성。(5) Masson 평분：B、C、E、F 조분별위(2.22±0.45)분、(2.44±0.3)분、(2.28±0.25)분、(2.44±0.35)분균명현대우 A 조(1.06±0.39)분、D 조(0.72±0.25)분，차이유통계학의의(P＜0.05)；B、C、E、F 조각조간평분량량비교，차이균무통계학의의(P＞0.05)；G 조평분(2.67±0.30)분교 B、E 조대，차이균유통계학의의(P＜0.05)；H 조평분최소，위(0.222±0.27)분。결론 BMPIII 여 BMPIV 량충활성다태인공복합재료균구유골유도능력；0.4 g / L 화0.8 g / L 농도골유도능력교0.2 g / L 농도대；0.4 g / L BMPI 골유도능력교 BMPIII 여 BMPIV 대，경괄우성위골유도재료。
Objective To evaluate osteoinductive potential of bone morphogenetic protein ( BMP ) III and BMPIV active polypeptides in animals by detecting ectopic bone formation ability of the 2 active polypeptides in different concentrations in rats, and to elect the best polypeptide in the best concentration by comparing with the osteoinductive activity of 0.4 g / L BMPI.Methods A total of 48 Sprague-Dawley ( SD ) rats were randomly divided into 8 groups ( groups A, B, C, D, E, F, G, H ), with 6 rats in each group. The composite materials of 0.2 g / L, 0.4 g / L and 0.8 g / L BMPIII and 0.2 g / L, 0.4 g / L and 0.8 g / L BMPIV with hydroxyapatite and poly lactic acid ( HA / PLA ) were implanted into superifcial muscles of the hips of the rats in group A, B, C, D, E and F respectively. The rats in the experimental control group ( group G ) received the composite materials of 0.4 g / L BMPI with HA / PLA and the rats in the blank control group ( group H ) only received HA / PLA. At 3 weeks after the operation, the anteroposterior X-ray films of the back were taken. The X-ray and CT images were observed at 5 weeks after the operation. The samples were taken out, stained by HE and Masson and observed under the light microscope at 3 and 5 weeks after the operation respectively. Using the method of nonparametric statistics, the osteogenic properties of the rats were analyzed at 5 weeks after the operation.Results All the rats underwent operation successfully, and their diets, activities and wound healing were ifne. All the rats were alive. The hardness of implantation materials was increased in each group. ( 1 ) According to the X-ray iflms, the following results were found. At 3 weeks after the operation, there were slightly fuzzy shadows in the implantation area, which were more obvious in group C and F. At 5 weeks after the operation, there were slight shadows in group A and D, which were more obvious in group C and F. There were no shadows in group H. ( 2 ) The CT images showed there were fuzzy low-density shadows in group A and D, which were more obvious in group B, C, E, F and G, and no shadows in group H. ( 3 ) A few osteoblasts were noticed in the porous materials and attached to the hole wall in group A, B, C, D, E, F and G at 3 weeks after the operation based on the HE staining results. Part of them were degraded at 5 weeks after the operation. There were more cartilage matrix and cells in group B, C, D, E, F and G. Little framework material was degraded in group H. ( 4 ) According to Masson staining, there was a little chondrogen around degradable materials in group A and D. The chondrogen in group B and C and group E and F was more than that in group A and group D respectively. The blue area was relatively small in group G. There was little chondrogen around degradable materials in group H. At 5 weeks after the operation, the blue area in group A, B, C, D, E, F and G was obviously larger than that at 3 weeks after the operation. There was little chondrogen around degradable materials in group H, only a little blue chondrogen was found. ( 5 ) The Masson scores in group B, C, E and F were obviously larger than that in group A and D, and the differences were statistically signiifcant (P<0.05 ). There were no statistically signiifcant differences when the Masson scores were compared between each 2 of group B, C, E and F (P>0.05 ). The Masson score in group G was larger than that in group B and F, and the differences were statistically significant ( P<0.05 ).Conclusions The artificial composite materials of BMPIII and BMPIV active polypeptides possess osteoinductive potential. The osteoinductive potential of 0.4 g / L and 0.8 g / L BMPIII and BMPIV is stronger than that of 0.2 g / L BMPIII and BMPIV. The osteoinductive potential of 0.4 g / L BMPI is stronger than that of BMPIII and BMPIV, which is the best osteoinductive material.
