CAJ | 학술논문

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.
목적 제비부재저방광점막하층거세포기질적각취당/납미간기린회석복합미구.방법 이용거오제-매법제비저방광점막하층거세포기질;유화-교련법제비부재거세포기질적각취당/납미간기린회석복합미구;응용소묘전경、X선연사의분석미구형태、성분;매련면역흡부법검측미구내전화생장인자β1(TGF-β1)、혈관내피생장인자(VEGF)함량급루적석방솔;새서람(MTT)법재1、3、5、7d검측복합미구세포독성.결과 복합미구형태균균、정구형,립경분포재(3.52 ～29.65) μm.X선연사현시:미구포함상술3충성분.미구내TGF-β1화VEGF적함량분별위(98.32±25.40)화(7.910 ±0.001) pg/g;량충세포인자적루적석방솔위8～10d내능유지95％적농도.미구침제액표명재료무세포독성.결론 부재저방광점막하층거세포기질적각취당/납미간기린회석복합미구,립경교착,8 ～10d내유지유효적생장인자농도,재료무세포독성.
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 composite microspheres' s leach liquor is not the cytotoxicity.