凝胶包埋BMP-2基因及振荡构建工程化骨的生物效应评估

目的 分析构建工程化骨的生物学行为,评估优化构筑工程化骨的方法. 方法 用人骨形态发生蛋白2腺病毒载体(Ad-BMP-2)转染兔脂肪基质细胞,nB-TCP/Cs/PCL支架接种及构建工程化骨,分A组(基因转染加静态培养),B组(凝胶加生长因子加振荡培养)、C组(凝胶加基因转染加振荡培养)和D组(凝胶加生长因子加静态培养);第1、4、8、12、16、20、24、28天,电镜、共聚焦显微镜观察,检测凋亡率、增殖、碱性磷酸酶活性及体内成骨分析.结果 C组细胞生长旺盛,细胞外基质丰富,凋亡率低于其它组(P＜0.05),增殖活力、碱性磷酸酶水平均高于其它组(P＜0.05),工程化骨发育成熟. 结论 凝胶包埋成骨基因与振荡模式为构建工程化骨的理想方法.

The evaluation of biological potential for construction of tissue-engineered bone by combination of oscillationg seeding with encapsulated BMP-2 gene

Objective To analyze the osteogenic potential of tissue-engineered bone in vitro,and to evaluate an effective way in constructing tissue-engineered bone.Methods Tissue-engineered bone was constructed in the following ways:ADSCs were transfected with BMP-2 gene plus static seeding and static culture (group A),ADSCs were encapsulated with gels containg BMP-2 plus oscillating seeding and static culture (group B),gels encapsulation with BMP-2 gene and oscillating seeding(group C),gels encapsulation with BMP-2 and static seeding(group D).SEM and CLM were used to observe cellular pattern,apoptosis rate,cell activity,ALP level were detected at 1,4,8,12,16,20,24,28 days.New bone formation were observed after transplantation of bone construct in nude mice. Results Under observation of SEM and CLM. new tissue showed more remarkable cell proliferation with abundant ECM in group C,compared with other groups,the apoptosis rate in group C was significantly lower(P＜0.05),and level of cell activity,ALP,DNA and OCN were significantly higher(P＜0.05). Conclusion Gels encapsulation of BMP-2 gene combined with oscillating seeding promotes the cell augmentation and improves the quality of tissue-engineered bone.