摘要
目的通过生物矿化方法构建具有骨引导和骨诱导能力的胶原电纺丝支架,对大鼠颅骨标准骨缺损进行引导性骨再生研究。方法采用电纺丝法制备猪Ⅰ型胶原支架,将其浸入模拟体液(SBF)中进行生物矿化,扫描电镜观察表面结构并进行理化分析。将矿化前、后的胶原电纺丝分别接种BMSCs,为矿化前组和矿化后组,并设立空白对照。进行生物毒性检测,并将胶原电纺丝植入大鼠背部皮下观察体内生物相容性和降解性。利用PCR、Western-blot、ALP及ARS染色等方法,检测体外成骨效果;利用大鼠颅骨缺损修复实验,检测生物矿化胶原电纺丝的体内骨引导和骨诱导能力。结果矿化4周后,胶原电纺丝表面矿化颗粒明显增多,TGA及ICP结果显示无机物尤其是钙磷元素含量也有所增加。生物毒性检测和异位植入实验结果证明,该材料具有良好的生物相容性和降解性。体外成骨检测显示,矿化后胶原电纺丝体外促成骨能力优于矿化前组和空白对照组。在大鼠颅骨缺损修复试验中,矿化后胶原电纺丝修复的颅骨在新骨生成面积、BV/TV、BS/TV、Tb.N、BMD和荧光标记新骨生成量方面显著优于空白对照组,BS/TV、BMD以及荧光标记新骨生成量显著优于矿化前胶原电纺丝修复的颅骨。结论生物矿化胶原电纺丝可有效促进BMSCs在材料表面的成骨分化,作为引导性骨再生支架能有效促进新骨形成,修复大鼠颅骨缺损。
Objective To construct a composite scaffold with osteoconductivity and osteoinductivity by biomineralization,and to conduct a guided bone regeneration study on the critical calvaria defect of rats.Methods The collagen scaffolds were fabricated out of the swine typeⅠcollagen by electrospinning and immersed in simulated body fluid(SBF),then the inorganic components of SBF were deposited in collagen scaffold by biomineralization.After scanning electron microscope observation and physical and chemical properties test,rat bone marrow mesenchymal stem cells were seeded on the scaffold material of biological collagen before and after mineralization as premineralization group and postmineralization group,blank control group was also set up.The biotoxicity was detected by Live/Dead staining,and the biocompatibility and biodegradability were tested after subcutaneous implantation into rat's dorsum.PCR,Western-blot,ALP and ARS staining were used to detect the effects on osteogenic differentiation of mesenchymal stem cells in vitro.The osteoconductivity and osteoinductivity of mineralized collagen scaffold in vivo was tested in rat craniofacial defect model.Results After 4 weeks of mineralization,the mineralized particles increased obviously.According to the results of TGA and ICP,the inorganic substances,especially the amounts of calcium and phosphorus were increased.The biotoxicity test and the results of subcutaneous implantation showed good biocompatibility and biodegradability of the material.The results of PCR,Western blot,ALP and ARS staining revealed similar results,the ability of collagen electrospinning after mineralization was superior to that of premineralization group and the blank control group.In the calvaria defect repair test of rats,the new bone formation area,the percent bone volume(bone volume/tissue volume,BV/TV),bone surface density(bone surface/tissue volume,BS/TV),the number of trabecular(Tb.N),bone mineral density(BMD),the new bone formation area of mineralized collagen reconstructed calvaria were better than those in blank control group.The bone surface density,bone mineral density,the new bone formation area of mineralized collagen reconstructed calvaria were also significantly superior to those of non-mineralized collagen reconstructed calvaria.Conclusion The mineralized collagen scaffold can effectively promote the osteogenic differentiation of BMSCs on the material surface,and serve as a guiding scaffold for bone regeneration to effectively promote the formation of new bone and repair the skull defect of rats.
作者
彭亦雩
王梓
袁庆越
谷平
毕晓萍
PENG Yiyu;WANG Zi;YUAN Qingyue;GU Ping;BI Xiaoping(Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology,Shanghai 200011,China)
出处
《组织工程与重建外科杂志》
2019年第2期65-71,共7页
Journal of Tissue Engineering and Reconstructive Surgery
基金
国家自然科学基金(81470662)
上海交通大学医学院高原高峰计划"研究型医师项目"(20161419)
关键词
胶原电纺丝
生物矿化
骨缺损
骨再生
Collagen electrospinning
Biomineralization
Bone defect
Bone regeneration
