CAJ | 학술논문

目的探讨构建一种新型人工活性真皮的可行性.方法组织块法培养幼儿包皮成纤维细胞；采用酶-去垢剂法制备人脱细胞羊膜(HAAM);双相法制备碱性成纤维细胞生长因子(bFGF)-明胶-壳聚糖缓释微球；缓释微球黏附于HAAM;bFGF基体式负载于HAAM,绘制药物缓释曲线；将第3～4代成纤维细胞培养于负载缓释微球的HAAM;扫描电镜观察HAAM、缓释微球的表面特征及缓释微球与HAAM的粘附情况；Western印迹法检测成纤维细胞中层粘连蛋白的表达.结果制备的HAAM为白色半透明状薄膜,有较高的孔隙率,空隙不规则,孔径大小为10～100 nm,无细胞毒性;bFGF-明胶壳聚糖缓释微球分散较均匀,呈球形,粒径均匀,球体表面比较光滑,载药率为20 ng/g,包封率为80.5％,体外药物缓释曲线显示药物控释效果良好；成纤维细胞在支架表面爬行生长良好,层粘连蛋白表达较对照组高.结论通过将成纤维细胞种植于负载bFGF-明胶-壳聚糖缓释微球的HAAM,有望制备出一种新型的人工活性真皮.
목적 탐토구건일충신형인공활성진피적가행성.방법 조직괴법배양유인포피성섬유세포；채용매-거구제법제비인탈세포양막(HAAM);쌍상법제비감성성섬유세포생장인자(bFGF)-명효-각취당완석미구；완석미구점부우HAAM;bFGF기체식부재우HAAM,회제약물완석곡선；장제3～4대성섬유세포배양우부재완석미구적HAAM;소묘전경관찰HAAM、완석미구적표면특정급완석미구여HAAM적점부정황；Western인적법검측성섬유세포중층점련단백적표체.결과 제비적HAAM위백색반투명상박막,유교고적공극솔,공극불규칙,공경대소위10～100 nm,무세포독성;bFGF-명효각취당완석미구분산교균균,정구형,립경균균,구체표면비교광활,재약솔위20 ng/g,포봉솔위80.5％,체외약물완석곡선현시약물공석효과량호；성섬유세포재지가표면파행생장량호,층점련단백표체교대조조고.결론 통과장성섬유세포충식우부재bFGF-명효-각취당완석미구적HAAM,유망제비출일충신형적인공활성진피.
Objective To discuss the possibility of constructing a new kind of active skin substi tutes.Methods The culture of dermal fibroblasts were isolated from foreskin of an infant; preparation of human acellular amniotic extracellular matrix was conducted through the disposition of detergent and enzyme.Surface structure porosity and pore size were detected through scanning electron microscopy.bFGF-chitosan gelatin microspheres were prepared.The features such as the size distribution,drug content,drug encapsulating and in vitro release were studied with scanning electron microscopy,laser grainsize analyzer and ELISA method.MTT test was performed to observe cell proliferation and evaluate the biocompatibility in vitro.Results The cellular layer of amniotic membrane was completely removed but did not damage the collogen scaffolds structure by the disposal of enzyme and detergent.Scanning electron microscopy showed that the scaffolds had criss-cross structure and high porosity,the pore size was irregular and varied from 10 nm to 100 nm.Scanning electron microscopy of bFGF- gelatin-chitosan microspheres surface structure showed that the microspheres had spherical structure,uniform size and smooth surface quality.Controlled release curve showed that the sudden release of bFGF was obvious.Conclusions A new type of active skin substitutes is prepared through culturing fibroblasts on HAAM loaded controlled-released basic fibroblast growth factor from chi tosan-gelatin microspheres.