新型可注射微球型多孔壳聚糖/羟基磷灰石支架的生物学性能研究  

Biological properties of a new injectable porous microspherical chitosan / hydroxyapatite scaffold

作　　者：彭志斌[1] 邵林[1] 牟鳄贤 杜大江[1] 刘真[3] 王强[1] 陈琳[1] 

机构地区：[1]哈尔滨医科大学附属第二医院骨一科,哈尔滨医学硕士研究生150086 [2]四川省肿瘤医院乳腺外科,成都610041 [3]哈尔滨医科大学附属第二医院科研实验中心,哈尔滨150086

出　　处：《医学研究生学报》2015年第11期1179-1184,共6页Journal of Medical Postgraduates

基　　金：黑龙江省自然科学基金(D201162);哈尔滨市科技创新人才研究专项资金项目(2012RFXXS049)

摘　　要：目的目前可注射组织工程骨或者载体支架多为凝胶状,其表面积、强度、硬度等方面无法满足复杂多变的骨、软骨缺损修复的要求。文中主要评价新型可注射微球型多孔壳聚糖/羟基磷灰石陶瓷支架的生物学性能。方法制备质量分数分别为30%、50%、70%的可注射多孔壳聚糖/羟基磷灰石复合微球,分别利用液氮的冷冻效应(液氮组)和非液氮途径(非液氮组)处理后烧结获得羟基磷灰石陶瓷球,拟做为新型组织工程骨的载体支架材料。通过扫描电镜观察支架的微细结构,测定孔隙率;通过生物力学实验测定力学性能。将人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVECs)种植于多孔壳聚糖/羟基磷灰石陶瓷支架,通过电镜扫观察HUVECs在多孔壳聚糖/羟基磷灰石陶瓷支架内部的生长情况验证多孔壳聚糖/羟基磷灰石陶瓷支架的生物融合性。结果液氮组和非液氮组不同质量分数的陶瓷支架肉眼无差异。扫描电镜下的外观图及内部图可见:2组微球陶瓷支架均呈球形、大小均匀、形态规则。液氮处理过的微球陶瓷支架表面见大量不规则孔隙;而非液氮处理微球陶瓷支架表面比较光滑,无明显的孔隙;2组微球陶瓷支架随着壳聚糖/羟基磷灰石的质量百分比的增大,内部的孔隙随之减少,使支架内部结构变的致密。液氮组50%质量分数支架平均直径、平均压缩强度均高于30%质量分数支架[(0.48±0.11)mm vs(0.40±0.08)mm,(1.75±0.14)MPa vs(1.21±0.12)MPa,P<0.05)],孔隙率低于30%质量分数支架[(79±2)%vs(87±1)%,P<0.05],平均孔径差异无统计学意义(P>0.05)。电镜扫描见HUVECs在多孔三磷酸钙支架内部呈多突触样附着,生长良好。结论液氮组50%质量分数多孔壳聚糖/羟基磷灰石陶瓷支架具有较强的力学强度和较好的生物融合性,能够做为新型组织工程骨的载体支架。Objective At present,the majority of injectable tissue engineering bones or carrier stents are gel,whose surface area,intensity,and hardness cannot satisfy the requirements of the repair of complex and varied bone and cartilage defects. This paper evaluated the new injectable microspherical porous chitosan / biological properties of the hydroxyapatite ceramic scaffold. Methods Injectable porous chitosan / hydroxyapatite composite microspheres with mass fractions of 30%,50%,and 70% were prepared respectively. The hydroxyapatite ceramic ball was obtained by sintering with liquid nitrogen freezing（ liquid nitrogen group） or without liquid nitrogen processing（ non- liquid nitrogen group） as a new carrier of bone tissue engineering scaffold material. The microstructure of the scaffold was observed and the porosity measured under the scanning electron microscope. The mechanical properties were determined through biomechanical experiments. Human umbilical vein endothelial cells（ HUVECs） were grown in the porous chitosan / hydroxyapatite ceramic scaffold followed by observation of the growth of the cells and validation of the biological fusion of the scaffold. Results No difference was observed with the naked eye in the ceramic scaffold of different mass fractions in the liquid nitrogen and non- liquid nitrogen groups.Scanning electron microscopy exhibited spherical shape,uniform size,and regular morphology of the ceramic scaffolds in both groups. A large number of irregular pores were seen in the surface of the microspherical ceramic scaffolds treated with liquid nitrogen but not in the surface of those not treated. With increased mass percentage of chitosan / hydroxyapatite,the internal pores were reduced and the interior structure compacted. In the liquid nitrogen group,the scaffold of 50% mass fraction had a significantly larger diameter（ [0. 48 ± 0. 11]mm）,higher compression intensity（ [1. 75 ± 0. 14]MPa）,and lower porosity（ [79 ± 2]%） than that of 30% mass fraction（ [0. 40 ±0. 08] mm,[

关 键 词：组织工程 骨再生 壳聚糖 羟基磷灰石 多孔支架 人脐静脉内皮细胞 

分 类 号：R318[医药卫生—生物医学工程]