磷酸钙骨水泥复合bFGF与VEGF修复兔桡骨缺损的实验研究
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摘要
目的观察分别混合碱性成纤维细胞生长因子(bFGF)、血管内皮生长因子(VEGF)和bFGF+VEGF的磷酸钙骨水泥(CPC)移植修复兔桡骨骨缺损的成骨作用,评价bFGF与VEGF单独及联合应用治疗骨缺损的效果。
方法选取36只健康家兔,体重为2.5～3.0kg。所有动物均建立双侧桡骨1.0cm的骨缺损实验模型。采用完全随机法将72侧前肢分为A、B、C、D四组,每组18侧。其中A组为对照组,实验组分为B、C、D三组。A组于骨缺损部位植入单纯CPC;B组植入复合bFGF的CPC;C组植入复合VEGF的CPC;D组植入联合应用bFGF和VEGF的CPC。分别于术后3、6、12周三个时间点切取标本,每组每个时间点根据随机原则取6个标本,通过X线摄片、骨密度测量、生物力学测试、组织形态学观察及评分等手段,观察各个时期新骨形成的情况。
结果
1、大体解剖观察显示12周时,D组骨缺损完全修复,B、C两组骨缺损亦基本修复,而A组仅有较多新骨形成,骨缺损未修复。
2、X线摄片显示12周时D组骨缺损完全修复,材料基本降解;B、C两组骨缺损基本修复。骨密度测量结果显示术后各时间点,实验组结果均高于对照组。术后12周时D组骨密度明显高于B、C两组(P＜0.05),B、C两组之间无明显差异(P＞0.05)。
3、生物力学测试(12周时)显示实验组最大抗折载荷明显高于对照组(P＜0.05)。D组明显高于B、C两组(P＜0.05);B、C两组之间无明显差异(P＞0.05)。
4、组织形态学观察显示在3周时,实验组骨小梁生长旺盛,有透明软骨形成;而对照组中骨小梁稀少。6周时,实验组中类骨组织转变为编织骨,形成骨性骨痂,且D组新骨量明显多于B、C两组。12周时,编织骨改建为成熟的板层骨,D组中皮质骨和髓腔的正常关系以及骨小梁正常的排列结构重新恢复,B、C两组则未见骨髓腔再通;A组中新骨仅少量增加。新骨形成及骨塑形评分显示术后各时间点D组均高于A、B、C三组(P＜0.05)。
结论
1、CPC/bFGF、CPC/VEGF和CPC/bFGF+VEGF三种复合物均能促进骨组织生长及加速骨缺损的修复;
2、CPC/bFGF+VEGF复合物促进骨组织生长的能力明显优于CPC/bFGF以及CPC/VEGF复合物,说明bFGF与VEGF促进骨生长具有协同作用。
Objective:To investigate the effect of calcium phosphate cement respectively mixed with bFGF,VEGF and both of them in repairing bone defects,and to evaluate their abilities of bFGF and VEGF to repair bone defects as separate or joint application.
Methods:A total of 36 rabbits(weight from 2.5 to 3.0kg) were randomly divided into four groups with 9 rabbits(18 sides of anterior limb) in each group,including goup A,B,C and D.For all rabbits 1.0 cm bone defects were created in both sides of radius.And then these bone defects region were implanted with corresponding composites:goup A was implanted with CPC only,goup B with CPC/bFGF,goup C with CPC/VEGF while goup D with bFGF and VEGF.At the 3~(rd),6~(th) and 12~(th) weeks after operation 6 specimens for each group were randomly selected. The effects were partly assessed by X-ray film examination,bone mineral density measurements,biomechanical test and histological observation.
Results
1.Gross anatomy observation showed that at the 12~(th) week the bone defcts in group D were completely repaired whereas these bone defects in goup B and C were just basically repaired.However there was a few of new bone formation in group A.
2.The X-ray showed that at the 12~(th) week the bone defcts in group D were completely repaired with CPC generally degraded,whereas these bone defects in goup B and C were just basically repaired.The BMD showed that at different time points the bone mineral density of experimental group was higher than the control group.At the 12~(th) week the bone mineral density of group D was significantly higher than group B and C(P＜0.05),while there was no distinct difference between these two groups(P＞0.05).
3.Biomechanical testing(at the 12~(th) week) showed that the maximum bending load of the experimental group were significantly higher than that of the control group(P＜0.05).Group D was superior to group B and group C(P＜0.05);whereas there was no significant difference between these two groups(P＞0.05).
4.Histological observation indicated that at the 3~(rd) week lots of trabecula and some hyaline cartilage could be seen in the experimental group.However,there were only a little of trabecula in the control group.At the 6~(th) week preosseous tissue had turned into woven bone in the experimental group,and the gross of new bone-formation in group D were more than group B and C.At the 12~(th) week,woven bone had become mature lamellar bone in group D.At the same time,the normal relation of cortical bone and marrow had resumed,and so did the normal structure of trabecula.However,the recanalizationthe of bone marrow cavity could not be seen in group B and C.And there were only a little of new bone-formation in group A.At different time points the histological score of group D was higher than group A,B and C(P＜0.05).
Conclusion
1.These three kinds of composite:CPC/bFGF,CPC/VEGF and CPC/ bFGF+VEGF can promote the growth of bone tissue and speed up the repair of bone defects.
2.The composite of CPC/bFGF+VEGF is significantly better than the other two composites in promoting the growth of bone tissue.And it indicates that bFGF and VEGF have a synergistic effect in promoting the growth of bone tissue.
方法选取36只健康家兔,体重为2.5～3.0kg。所有动物均建立双侧桡骨1.0cm的骨缺损实验模型。采用完全随机法将72侧前肢分为A、B、C、D四组,每组18侧。其中A组为对照组,实验组分为B、C、D三组。A组于骨缺损部位植入单纯CPC;B组植入复合bFGF的CPC;C组植入复合VEGF的CPC;D组植入联合应用bFGF和VEGF的CPC。分别于术后3、6、12周三个时间点切取标本,每组每个时间点根据随机原则取6个标本,通过X线摄片、骨密度测量、生物力学测试、组织形态学观察及评分等手段,观察各个时期新骨形成的情况。
结果
1、大体解剖观察显示12周时,D组骨缺损完全修复,B、C两组骨缺损亦基本修复,而A组仅有较多新骨形成,骨缺损未修复。
2、X线摄片显示12周时D组骨缺损完全修复,材料基本降解;B、C两组骨缺损基本修复。骨密度测量结果显示术后各时间点,实验组结果均高于对照组。术后12周时D组骨密度明显高于B、C两组(P＜0.05),B、C两组之间无明显差异(P＞0.05)。
3、生物力学测试(12周时)显示实验组最大抗折载荷明显高于对照组(P＜0.05)。D组明显高于B、C两组(P＜0.05);B、C两组之间无明显差异(P＞0.05)。
4、组织形态学观察显示在3周时,实验组骨小梁生长旺盛,有透明软骨形成;而对照组中骨小梁稀少。6周时,实验组中类骨组织转变为编织骨,形成骨性骨痂,且D组新骨量明显多于B、C两组。12周时,编织骨改建为成熟的板层骨,D组中皮质骨和髓腔的正常关系以及骨小梁正常的排列结构重新恢复,B、C两组则未见骨髓腔再通;A组中新骨仅少量增加。新骨形成及骨塑形评分显示术后各时间点D组均高于A、B、C三组(P＜0.05)。
结论
1、CPC/bFGF、CPC/VEGF和CPC/bFGF+VEGF三种复合物均能促进骨组织生长及加速骨缺损的修复;
2、CPC/bFGF+VEGF复合物促进骨组织生长的能力明显优于CPC/bFGF以及CPC/VEGF复合物,说明bFGF与VEGF促进骨生长具有协同作用。
Objective:To investigate the effect of calcium phosphate cement respectively mixed with bFGF,VEGF and both of them in repairing bone defects,and to evaluate their abilities of bFGF and VEGF to repair bone defects as separate or joint application.
Methods:A total of 36 rabbits(weight from 2.5 to 3.0kg) were randomly divided into four groups with 9 rabbits(18 sides of anterior limb) in each group,including goup A,B,C and D.For all rabbits 1.0 cm bone defects were created in both sides of radius.And then these bone defects region were implanted with corresponding composites:goup A was implanted with CPC only,goup B with CPC/bFGF,goup C with CPC/VEGF while goup D with bFGF and VEGF.At the 3~(rd),6~(th) and 12~(th) weeks after operation 6 specimens for each group were randomly selected. The effects were partly assessed by X-ray film examination,bone mineral density measurements,biomechanical test and histological observation.
Results
1.Gross anatomy observation showed that at the 12~(th) week the bone defcts in group D were completely repaired whereas these bone defects in goup B and C were just basically repaired.However there was a few of new bone formation in group A.
2.The X-ray showed that at the 12~(th) week the bone defcts in group D were completely repaired with CPC generally degraded,whereas these bone defects in goup B and C were just basically repaired.The BMD showed that at different time points the bone mineral density of experimental group was higher than the control group.At the 12~(th) week the bone mineral density of group D was significantly higher than group B and C(P＜0.05),while there was no distinct difference between these two groups(P＞0.05).
3.Biomechanical testing(at the 12~(th) week) showed that the maximum bending load of the experimental group were significantly higher than that of the control group(P＜0.05).Group D was superior to group B and group C(P＜0.05);whereas there was no significant difference between these two groups(P＞0.05).
4.Histological observation indicated that at the 3~(rd) week lots of trabecula and some hyaline cartilage could be seen in the experimental group.However,there were only a little of trabecula in the control group.At the 6~(th) week preosseous tissue had turned into woven bone in the experimental group,and the gross of new bone-formation in group D were more than group B and C.At the 12~(th) week,woven bone had become mature lamellar bone in group D.At the same time,the normal relation of cortical bone and marrow had resumed,and so did the normal structure of trabecula.However,the recanalizationthe of bone marrow cavity could not be seen in group B and C.And there were only a little of new bone-formation in group A.At different time points the histological score of group D was higher than group A,B and C(P＜0.05).
Conclusion
1.These three kinds of composite:CPC/bFGF,CPC/VEGF and CPC/ bFGF+VEGF can promote the growth of bone tissue and speed up the repair of bone defects.
2.The composite of CPC/bFGF+VEGF is significantly better than the other two composites in promoting the growth of bone tissue.And it indicates that bFGF and VEGF have a synergistic effect in promoting the growth of bone tissue.
引文
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