[目的]观察重组h BMP-7转染ADSCs分化成骨修复兔股骨头缺血性坏死(avascular necrosis of the femoral head,ANFH)的疗效。[方法]设计和合成引物通过PCR扩增h BMP-7基因。取4只3个月龄新西兰大白兔颈背部皮下脂肪组织分离脂肪细胞并培...[目的]观察重组h BMP-7转染ADSCs分化成骨修复兔股骨头缺血性坏死(avascular necrosis of the femoral head,ANFH)的疗效。[方法]设计和合成引物通过PCR扩增h BMP-7基因。取4只3个月龄新西兰大白兔颈背部皮下脂肪组织分离脂肪细胞并培养传代。将h BMP-7经脂质体介导转染3代脂肪干细胞。36只新西兰大白兔ANFH模型随机分为3组。A组为模型对照组,B组为模型对照组处理的基础上减压后植入空质粒ADSCs;C组为模型对照组处理的基础上减压后植入重组h BMP-7真核表达载体转染的ADSCs。观察股骨头坏死模型兔的一般情况,并分别于术后4、8周每组处死6只行骨密度检查,收集到的模型股骨头标本分别行X线检查、HE染色及免疫组化检测BMP-7表达,检测ANFH修复重建的效果。[结果]术后第4周三组股骨近端骨密度变化分别为A组骨质疏松,B组和C组均为骨量减少,C组较B组骨质情况稍好但差异无统计学意义。术后第8周,三组骨密度C>B>A,差异有统计学意义。其他:4周时,X线检查,组织学检查A,B,C组的修复效果分别为C及B>A,但C、B两组间差异无统计学意义。BMP-7免疫组化的表达量为C>B>A。8周时,组织学检查A,B,C组的修复效果分别为C>B>A,BMP-7免疫组化的表达量也为C>B>A。[结论]经过h BMP-7转染修饰的ADSCs植入坏死的股骨头内成骨能力及修复坏死区域的功能较单独ADSCs强,具有应用于临床修复股骨头坏死的理论基础。展开更多
Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous s...Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous study showed that nanotechnology improved adipose-derived stem cell （ADSC） therapy for erectile dysfunction of cavernous nerve injury （CNI） by attracting cells in the corpus cavernosum. These results indicated the possibility of using a reduced dosage of ADSCs for intracavernous injection. In this exploratory study, we used lower dosage （2 × 105 cells） of ADSCs for intracavernous injection （ICI） and the nanotechnology approach. Intracavernous pressure and mean arterial pressure were measured at day 28 to assess erectile function. The low-dose ADSC therapy group showed favorable treatment effects, and nanotechnology further improved these effects. In vivo imaging of ICI cells revealed that the fluorescein signals of NanoShuttle-bound ADSCs （NanoADSCs） were much stronger than those of ADSCs at days 0, 1, and 3. Both immunofluorescence and Western blot analysis showed a significant increase in smooth muscle, endothelium, and nerve tissue in the ADSC group compared to that in the CNI group; further improvement was achieved with assisted nanotechnology. These findings demonstrate that nanotechnology can be used to further improve the effect of small dosage of ADSCs to improve erectile function. Abundant NanoADSCs remain in the corpus cavernosum in vivo for at least 3 days. The mechanism of erectile function improvement may be related to the regeneration of the smooth muscle, endothelium, and nerve tissues.展开更多
基金This research was supported by the Beijing Natural Science Foundation （Grant No. 7174362） and the National Natural Science Foundation of China （Grant No. 81601272）.
文摘Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous study showed that nanotechnology improved adipose-derived stem cell （ADSC） therapy for erectile dysfunction of cavernous nerve injury （CNI） by attracting cells in the corpus cavernosum. These results indicated the possibility of using a reduced dosage of ADSCs for intracavernous injection. In this exploratory study, we used lower dosage （2 × 105 cells） of ADSCs for intracavernous injection （ICI） and the nanotechnology approach. Intracavernous pressure and mean arterial pressure were measured at day 28 to assess erectile function. The low-dose ADSC therapy group showed favorable treatment effects, and nanotechnology further improved these effects. In vivo imaging of ICI cells revealed that the fluorescein signals of NanoShuttle-bound ADSCs （NanoADSCs） were much stronger than those of ADSCs at days 0, 1, and 3. Both immunofluorescence and Western blot analysis showed a significant increase in smooth muscle, endothelium, and nerve tissue in the ADSC group compared to that in the CNI group; further improvement was achieved with assisted nanotechnology. These findings demonstrate that nanotechnology can be used to further improve the effect of small dosage of ADSCs to improve erectile function. Abundant NanoADSCs remain in the corpus cavernosum in vivo for at least 3 days. The mechanism of erectile function improvement may be related to the regeneration of the smooth muscle, endothelium, and nerve tissues.
