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纳米生物陶瓷人工听骨的实验研究

The Study on the Ossicular Prosthesis of Nano-Bioactive Ceramics in Guinea Pigs

【作者】 吴照红

【导师】 刘立中;

【作者基本信息】 新疆医科大学 , 耳鼻咽喉科学, 2009, 硕士

【摘要】 目的:制备纳米多孔羟基磷灰石(HA)陶瓷支架;研究新型陶瓷支架的生物相容性。方法:用水热法法制备纳米级HA粉体,有机泡沫浸渍法制备多孔陶瓷支架,观察其形貌特征,组成成份及力学性能。采用鼠成骨细胞(periosteal-derived osteoblast cells,POB)与两种陶瓷支架的体外复合培养,观察细胞在材料上的生长及功能表达。本研究通过不同理化方法的加工工艺,制备纳米级的多孔羟基磷灰石(HA)支架,采用与鼠骨膜成骨细胞的体外复合培养,体内动物植入实验,评价多孔的HA和AO,这两种骨组织工程支架材料的生物相容性。论文研究纳米多孔HA陶瓷的制备及性能测试并对纳米多孔HA陶瓷的生物相容性进行评价。采用纳米HA颗粒悬液直接作用于成骨前体细胞。运用四吟盐比色法、碱性磷酸酶的方法分析悬液对成骨前体细胞增殖和分化影响。探讨了钙离子溶出的培养液和条件培养液吸附的对细胞增殖、分化的影响。在体内行豚鼠听泡内植入实验。选择健康、白色豚鼠30只(60耳),采用耳后进路,听泡后外侧骨壁钻孔,左侧听泡植入AO普通陶瓷块作为对照组,右侧听泡植入纳米羟基磷灰石(HA)复合多孔陶瓷人工听骨做为实验组,分别于术后1、2、4、8、12、16周处死,行植入物表面接触缘大体形态学及扫描电镜观察。结果:制备出具有良好孔隙结构和一定强度的纳米多孔陶瓷支架材料;豚鼠POB在HA和AO表面良好的生长,并发挥成骨功能。30只豚鼠术后均无感染,听泡光镜及扫描电镜检查,植入物界面形态良好,结构完整,边界清晰。4周时,材料周围被薄层纤维组织包裹,因轻微炎性反应,有少量巨噬细胞和多核巨细胞存在,有纤维组织长入生物陶瓷边缘区域。12周时,均无炎性反应,可见大量胶原纤维组织长入陶瓷中央。纳米HA复合多孔陶瓷可见少量的骨组织。术后16周,有新骨形成,而对照耳仅有粘膜覆盖,无新骨形成。结论:以HA的纳米级粉体为原料,采用有机泡沫浸渍法可制备出具有良好孔隙结构和较好力学强度的多孔陶瓷支架;新型多孔陶瓷支架有良好的生物相容性。

【Abstract】 Objective:To make porous nano-bioceramics and to study their biocompatibility as bone tissue engineerings caffolds.Methods:The nanometer powders of hydroxyapatite(HA) and tricalcium phosphate(AO) were prepared by Sol-gel methods and chemical deposition methods and the porous nano-bioceramics were made by foam sockage technics. The biocompatibility of new porous nano- bioceramics were evaluated by co-culture with mice periosteal-derived osteoblast cells (POB) in vitro. The attachment and the growth of POB on the scaffolds were investigated by morphologic, proliferative abilities Osteogenic activity. In the animal experiment nano-HA prostheses were put in the right ateralbullae as ossicular prothesis of the ears of 30 healthy white guinea pigs (totally 60 ears) , AO ceramic was in the left as the control group .By retroauricular approach ,a hole was made in poster lateral bone wall of the acoustic vesicle . Each group animals executing on 1 . 2 . 4 . 8 . 12 . 16 weeks after operation ,investigated responses of tissue to materials by general and common histology. Results:HA and AO porous nano-bioceramics scaffolds had a good network pore system and mechanic properties. POB can fully attached to, extended on both HA and AO, and excreted extracellular matrix. The nano-HA prostheses taken out from 60 ears of the guinea pigs were normal and complete structure, and clear margin in gross anatomy and microscopically. In each case a layer of fibrous tissue surrounding the frame work ,a moderate inflammatory response and foreign body reaction were observed at 4 weeks after surgery. At 12 weeks ,the inflammatory response disappeared . Dense fibrous tissue was grown into the center of the bioceramic . . A little bone tissue growed into the nano-bioceramic. the new bone was found binding with the grafted material closely in group of nano-HA in 16 weeks. In the group of control, the new bone tissue were not found. Conclusion:With the nanometer powders of HA and AO, the porous nano-bioceramics can be produced by foam sockage technics. The new porous nano-bioceramics have good biocompatibility with POB.

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