负载去细胞基质的壳聚糖/纳米羟基磷灰石复合微球的制备及其性状  

Production and characterization of loadedly decellularizated matrix＇s chitosan/nanohydroxyapatite composite microspheres

作　　者：乔瑞红[1] 王东[1] 孙海钰[1] 高敏[1] 

机构地区：[1]山西医科大学第二医院骨科,太原030001

出　　处：《中华实验外科杂志》2014年第8期1778-1780,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 制备负载猪膀胱黏膜下层去细胞基质的壳聚糖/纳米羟基磷灰石复合微球.方法 利用去污剂-酶法制备猪膀胱黏膜下层去细胞基质;乳化-交联法制备负载去细胞基质的壳聚糖/纳米羟基磷灰石复合微球;应用扫描电镜、X线衍射仪分析微球形态、成分;酶联免疫吸附法检测微球内转化生长因子β1（TGF-β1）、血管内皮生长因子（VEGF）含量及累积释放率;噻唑蓝（MTT）法在1、3、5、7d检测复合微球细胞毒性.结果 复合微球形态均匀、呈球形,粒径分布在（3.52 -29.65） μm.X线衍射显示：微球包含上述3种成分.微球内TGF-β1和VEGF的含量分别为（98.32±25.40）和（7.910 ±0.001） pg/g;两种细胞因子的累积释放率为8-10d内能维持95％的浓度.微球浸提液表明材料无细胞毒性.结论 负载猪膀胱黏膜下层去细胞基质的壳聚糖/纳米羟基磷灰石复合微球,粒径较窄,8 -10d内维持有效的生长因子浓度,材料无细胞毒性.Objective The loadedly porcine urinary bladder mucous membrane＇s decellularizated matrix＇ s chitosan/nanohydroxyapatite composite microspheres （DM＇ S CS/nHA） is to be prepared.Methods The detergent-enzyme digestion method was used to produce decellularizated matrix; W/O combined with multiple emulsification chemical crosslink technique was used to prepare for the loadedly decellularizated matrix＇ s chitosan/nanohydroxyapatite composite microspheres.We observed the microspheres＇s characterization of the surface shape and diameter distribution by scanning electron microscope （SEM） ; The composite microspheres＇ components were analysed by the X-ray diffraction （XRD） examination.The contents of the dissolvable protein as to transforming growth factor （TGF）-β1 and vascular endothelial growth factor （VEGF） in the composite microspheres were measured by enzyme linked immunosorbent assay （ELISA）,and the cumulative release rate of both TGF-β1 and VEGF was also surveyed; The cytotoxicity for the microspheres＇s leach liquor was researched in 1,3,5,7 d by methyl thiazol tetrazolium （MTT） method.Results The loadedly decellularizated matrix＇s chitosan/nanohydroxyapatite composite microspheres were spherical in shape,and the particle diameter ranged from 3.52 μm to 29.65 μm.XRD analysis demonstrated that the composite microspheres contained DM,CS,nHA.The contents of the TGF-β1 and VEGF in the decellularizated matrix were （98.32 ± 25.40） pg/g; TGF-β1：（7.910 ± 0.001） pg/g respectively,and the cumulative release rate of both TGF-β1 and VEGF was that the 95％ effective concentration could maintain for 8-10 days.The cytotoxicity experiments for the microspheres＇s leach liquor demonstrated no cell toxicity.Conclusion The loadedly Porcine urinary bladder mucous membrane＇s decellularizated matrix＇ s chitosan/nanohydroxyapatite composite microspheres exhibit narrow particle diameter distribution.They can maintain TGF-β1 and VEGF concentration within 8-10 days.The com

关 键 词：去细胞基质 壳聚糖 纳米羟基磷灰石 复合微球 制备 

分 类 号：R318.08[医药卫生—生物医学工程]