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异种脱蛋白骨复合物修复大段骨缺损的血管化观察
背景: 异种骨修复动物大段骨缺损过程中血管化的报道较少.目的: 验证异种脱蛋白骨作为组织工程支架材料修复长骨大段缺损成骨过程中的血管化特点.设计、时间及地点: 随机对照动物实验,于2005-03/2007-02在解放军第三军医大学完成.材料: 山羊24只,体质量(22.5±2.5)kg,右侧胫骨中下段截除胫骨总长度20%形成节段性骨缺损.方法: 24只实验山羊随机分为2组,实验组16只动物植入异种脱蛋白骨+自体间充质干细胞+重组人骨形态发生蛋白2,用半环槽式外固定器固定;以另8只植入自体骨为对照.术后每隔4周进行股动脉墨汁灌注后,取新骨组织制成厚切片.主要观察指标: ①大体解剖及影像学观察新骨血管化情况.②厚切片观察新骨血管网,并用Image-proplus真彩图像分析软件测量血管数目和面积.结果: 所有实验山羊术后伤口无溃烂、流脓,无死亡.①动物肌肉和软组织已经被墨汁黑染,皮下、肌肉筋膜和骨膜可见血管大小、分支、走行方向及相互吻合呈网状.术后4周骨缺损区出现植入物边缘毛糙,8周出现大小不等、形状不规则的透光性骨吸收区.12周以后,两组骨缺损区两断端之间高密度钙化影,新生骨与两断端完全相连接.②术后4～24周,实验组与对照组血管表现数目逐渐增多,排列渐趋规则,血管大小、分布区域趋向均匀.两组血管数目和面积相比,差异均无显著性意义(P>0.05).结论: 异种脱蛋白骨复合物材料修复大动物大块骨缺损过程中血管化程度与自体骨相当.
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改良法制备异种脱蛋白骨复合重组人骨形态发生蛋白支架修复长骨大段骨缺损
BACKGROUND: For its extensive sources and specific biological characteristics, heterogeneous deproteinized bone (DPB) as the tissue-engineered scaffold material gets a good application prospect in repair of large segmental bone defects.OBJECTIVE: To prepare scaffold material by compositing modified heterogeneous DPB with recombinant human bone morphogenetic protein 2 (rhBMP2) and evaluate the osteogenic ability in repairing large segmental long bone defects in large-sized animals.METHODS: Twenty-four goats were randomly and evenly divided into three groups: DPB, autologous bone, and DPB+rhBMP2. Bone defects (20% in length) were made in the middle and lower segments of the right tibia of each goat, and according to group design,different bone grafts were implanted and fixed using semi-ring slot external fixer. At 4-24 weeks after surgery, X-ray examination was performed once every other 4 weeks. At 24 weeks after surgery, newly formed bones were taken out for determining the repair effects of bone defects through duat-energy X-ray analysis, histological observation, and biomechanical analysis.RESULTS AND CONCLUSION: The prepared DPB exhibited a three-dimensional porous structure, with a porosity of (78.5±6.45)% and pore size of (472.5±7.02) μm. Time-dependent bone repair was found in each group. Results regarding anti-compression test, three-point anti-bend test, and anti-torsion test were greatest in the autologous bone group, followed by the DPB+rhBMP2 group, and lastly the DPB group. There were no significant differences in tibial bone density, bone mineral content,anti-compression pressure and ultimate pressure, anti-bending load and ultimate toad, anti-torsion torque and ultimate torque between autologous bone group, DPB+rhBMP2 group and normal bone control group. These findings indicate that modified heterogeneous DPB composited with rhBMP2 has osteogenic ability equivalent to autologous bone in repairing large segmental long bone defects in goat tibias. Therefore, it can be applied as tissue-engineered scaffold material.
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改良法制备异种脱蛋白骨复合重组人骨形态发生蛋白支架材料修复长骨大段骨缺损
背景:异种脱蛋白骨因其来源广泛和独特的生物学特性作为组织工程骨支架材料而受到较多关注.目的:采用改良法制备异种脱蛋白骨复合重组人骨形态发生蛋白支架材料,评价其修复大动物大段长骨缺损的成骨能力.设计、时间及地点:随机对照动物实验,于2005-05/2006-12在解放军第三军医大学实验动物中心及西南医院中心实验室完成.材料:市售猪股骨;山羊24只,10～12个月龄,雌雄不限.半环槽外固定器由解放军第三军医大学李起鸿教授研制.专利号:CN01256565.方法:山羊24只,随机分为单纯异种脱蛋白骨组、自体骨组及异种脱蛋白骨+重组人骨形态发生蛋白组,在每只山羊右侧胫骨中下段截除胫骨总长度20%形成节段性骨缺损,按分组情况植入不同材料,采用半环槽式外固定器固定.异种脱蛋白骨制备参照文献完成.主要观察指标:术后4～24周每隔4周进行X射线及术后24周取新生骨进行双能X射线、组织学、生物力学检测骨缺损修复效果.结果:①制得的异种脱蛋白骨为三维多孔结构,其孔隙率为(78.5±6.45)%,孔径大小为(472.5±7.02)μ m.②各组骨缺损表现为时间依从性的骨修复.生物力学试验抗压缩、三点抗弯曲、抗扭转试验,以及同一时间点平均成骨量均为自体骨组>异种脱蛋白骨+重组人骨形态发生蛋白组>单纯异种脱蛋白骨组.③自体骨组、异种脱蛋白骨+重组人骨形态发生蛋白组与正常组比较,山羊胫骨骨密度及骨矿含量、抗压缩压强及极限压强、抗弯曲载荷及极限载荷、抗扭转扭矩及极限扭矩平均值差异均无显著性意义.结论:改良法制备的异种脱蛋白骨复合重组人骨形态发生蛋白修复山羊胫骨大段缺损成骨能力与自体骨相当,可以作为组织工程支架材料.
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脱蛋白异种骨复合富血小板血浆修复股骨干骨缺损
目的 将富血小板血浆与异种脱蛋白骨结合,观察其修复股骨干骨缺损的疗效.方法 2次离心取得患者自体富血小板血浆,术前将其与异种脱蛋白松质骨混合,植于股骨干骨折骨缺损处.对照组仅用异种脱蛋白松质骨植骨.术后7 d和1、2、3、6、12个月定期摄X线片检查,参照Lane-Sandhu的X线评分标准评分,并比较两组骨痂灰度,比较两组愈合情况及愈合速度.结果 两组Lane-Sandhu的X线评分在术后1、2、3、6个月时,差异有显著性(P＜0.05);而在术后12个月,两组差异无显著性(P＞0.05).骨痂灰度术后1、2、3、6个月时两组差异有显著性(P＜0.05).结论 富血小板血浆复合异种脱蛋白松质骨可促进和加速骨缺损的修复.
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异种脱蛋白骨复合物修复大段骨缺损的血管化观察
[目的] 探讨异种脱蛋白骨(heterogeneous deprotein bone,HDPB)复合物修复胫骨大段骨缺损过程中的血管化实验研究,为异种脱蛋白骨的临床应用提供实验依据.[方法] 山羊24只,在右侧胫骨中下段截除胫骨总长度20%形成节段性骨缺损,实验组18只,植入异种DPB+自体MSCs+rhBMP2,用半环槽式外固定器固定.于术后每隔4周对3只进行股动脉墨汁灌注后处死,取新骨组织制成厚切片观察血管化情况,以另6只植入自体骨为对照.[结果] 术后4～24周,实验组与对照组血管表现数目逐渐增多,排列渐趋规则,血管大小、分布区域趋向均匀.测量血管数目和面积,结果显示4～24周2组血管数目和面积统计学分析差异无显著性意义(P＞0.05).[结论] 异种脱蛋白骨复合物修复大动物大块骨缺损过程中血管化程度与自体骨相当,可以作为组织工程支架材料,并对其免疫原性和成骨能力等性能进行进一步研究.
