骨髓间充质干细胞复合多孔纳米羟磷灰石/聚酰胺6整复大鼠颅骨缺损的实验研究  

Reconstruction of critical sized calvarial defects by porous nano-hydroxyapatite/polyamide 6 composite with bone marrow mesenchymal stem cells in rat

作　　者：高莺[1,2] 李继华[1] 李玉宝[3] 左奕[3] 胡静[1] 马永清[1] 王雪梅[1] 

机构地区：[1]口腔疾病研究国家重点实验室、四川大学,四川成都610041 [2]山西医科大学第一医院口腔科,山西太原030001 [3]四川大学纳米生物材料中心,四川成都610041

出　　处：《华西口腔医学杂志》2010年第1期17-20,24,共5页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目(30400506)

摘　　要：目的初步探讨纳米羟磷灰石/聚酰胺6(n-HA/PA6)多孔材料对体外培养的大鼠骨髓间充质干细胞(BM-SCs)增殖及分化的影响,以及BMSCs作为种子细胞、多孔n-HA/PA6作为支架材料构建组织工程化骨修复大鼠颅骨极限骨缺损的可行性及整复效果。方法矿化诱导的第3代BMSCs与n-HA/PA6多孔材料复合培养,MTT检测细胞增殖,ALP染色检测骨向分化。将BMSCs与n-HA/PA6复合物植入大鼠颅骨8mm骨缺损处,4、8、16周时,应用组织学、扫描电镜观察植入物与骨组织交界处的成骨修复情况,并与单纯n-HA/PA6植入组修复效果进行比较。结果与n-HA/PA6复合培养的BMSCs生长良好,细胞增殖未受影响,ALP染色阳性。BMSCs复合n-HA/PA6植入4周时,有较多新骨长入支架孔穴;8周时,材料和宿主骨融为一体,接近正常骨;16周时,材料和天然骨形成骨性结合。单纯n-HA/PA6组植入4周时,新骨形成较少;8周时,新骨明显增加,但骨钙化程度较低;16周时,2组无明显差异。结论多孔n-HA/PA6支架材料对种子细胞BMSCs的增殖和骨向分化无影响;BMSCs作为种子细胞、n-HA/PA6多孔复合体作为支架材料构建组织工程化骨能够加速界面骨愈合,有效修复颅骨缺损,具有潜在的骨组织工程应用前景。Objective To evaluate the effects of the nano-hydroxyapatite/polyamide 6（n-HA/PA6） on the proliferation and osteoblastic differentiation of rat bone marrow mesenchymal stem cells（BMSCs）,and the feasibility of using both for constructing tissue engineered bone in the calvarias of rats with critical sized defects.Methods The third passage of BMSCs were cultured in osteoblastic medium and seeded on the scaffolds of n-HA/PA6,the proliferation of the BMSCs was tested by MTT（3-{4,5-dimethylthiazol-2yl}-2,5-diphenyl-2H-tetrazolium-bromide） on scheduled dates,and the osteoblastic differentiation of the BMSCs were measured by alkaline phosphatase（ALP） staining.Furthermore,the scaffolds with or without BMSCs in rat calvarial defects,after 4 weeks,8 weeks,and 16 weeks have been implanted.Histology and scanning electron microscope were used to test the bone healing in the different groups.Results The BMSCs seeded on the n-HA/PA6 grew well,the proliferation of cells was not affected by the scaffold,and the staining of ALP was also positive.At 4 week and 8 week after implantation,the n-HA/PA6 with BMSCs showed more new bone formation on the surface of scaffolds,with a better osseointegration of implant and host bone when compared with the group of n-HA/PA6 without BMSCs.However,there was no significant difference between these two groups at 16 week.Conclusion The porous n-HA/PA6 has no negative effects on the proliferation and osteoblastic differentiation of rat BMSCs,and using BMSCs as seed cells and n-HA/PA6 as scaffolds is a good choice for constructing tissue engineered bone due to the enhanced new bone formation and osseointegration.

关 键 词：骨髓间充质干细胞 纳米羟磷灰石/聚酰胺6 颅骨缺损 

分 类 号：R651[医药卫生—外科学]