以京尼平交联制备新型人工神经支架材料及其生物学特性的对比研究  

Genipin crosslinked preparation of new peripheral nerve tissue engineered scaffolds and comparison of their biological characteristics

作　　者：李沫[1] 吴邦耀[1] 胡学昱[1] 黄景辉[1] 张永光[1] 罗卓荆[1] 

机构地区：[1]第四军医大学附属两京医院骨科,西安710032

出　　处：《中华创伤杂志》2010年第2期165-171,共7页Chinese Journal of Trauma

基　　金：国家高技术研究发展计划资助项目（20024A216101）

摘　　要：目的从超微结构、孔隙率、溶胀率、降解率、交联度及细胞毒性等方面分别比较紫外线、京尼平及戊二醛交联后的壳聚糖复合I型胶原蛋白人工神经支架材料的生物学特性。方法（1）材料按交联方法不同分为二三组：紫外线组、京尼平组、戊二醛组。（2）经扫描电镜观察三组材料内部结构的排列规律及走形，测量其孔径大小、计算孔隙率及孔径分布等指标：（3）溶胀率与体外降解率：二三组材料交联完后市即称重（W0），然后存培养皿中力11入10ml无菌PBS，24h后用无菌滤纸擦干水分称重（W1）：溶胀牢（％）=（w1-W0）／W0×100％。剩余材料于4，8，12用分别取出后称重（W2）.降解率（％）=（W1-W2）／W1×100％。（4）检测交联度：每组取10根材料，其巾5根加入碳酸氢钠和二硝基苯磺酸（TNBS），再加盐酸，往346Dill测吸光度（A）值（A三硝基苯磺酸）。另外5根先加盐酸，然后再加TNBS，其余步骤相同，测得吸光度取平均值作为对照（A对），交联后吸光度值为：A交联后n=A三硝基苯磺酸-A对。再取一组10根术交联过的材料，以同样步骤测吸光度，得到交联前吸光度（A交联前）。交联度=（A交联前-A交联后）／A交联前x100％。（5）细胞毒性试验：遵照GB／T16886／ISO10993医疗器械生物学评价之休外细胞毒性试验原则，采用国际标准的两种试验方法，选用建系的1929小鼠成纤维细胞对改性后的支架材料进行体外细胞毒性试验。结果（1）未交联的材料为均匀圆柱状，内鄙为孑L径均匀且平行排列的微观结构，其微孔直径为30～120μm；交联后紫外线组孔径基本小变，京尼平、戊二醛两组孔径均变小。（2）京尼平和戊二醛组孔隙率差异无统计学意义，两者都要高于紫外线绀；而溶胀率京尼平缉高于戊二醛组，戊二醛组又高于紫外线组：（3）京尼平和戊二醛两�Objective To compare biological properties of chitosan composite artificial neural type I eoltagen scaffold material cross-linked with ultraviolet rays （UV） , genipin （GP） and glutaralde- hyde （GTA） in aspects of uhrastmcture, porosity, swelling rate, degradation rate, erosslinking degree and cytotoxicity. Methods （ 1 ） According to different cross-linking methods, biomaterials were divided into three groups, ie, UV group, GP group and GTA group. （2）The microstructure of three groups was observed under scanning electron microscope （SEM） to measure pore size, porosity rate and pore-size distribution. （3）Swelling rate and in vitro degradation rate： the biomaterials were weighed （W0 ） after crosslinking and then immersed in culture medium containing 10 ml aseptic phosphate buffer solution （PBS）. The samples were drawn from the culture medium after 24 hours, wiped with filter paper to re- move excess liquid and weighed （ Wl ）. Swelling rate（ % ） = W1 - W0/W0 x 100%. The remaining sam- ples from each group were weighed （W2 ） at 4, 8, 12 weeks with the same procedure. Degradation rate （ % ） = W1- W2/W1 x 100%. （4） Determination of cross-linking index： 10 samples were prepared from each group, five samples from which were reacted with trinitro-benzen-sulfonic acid （TNBS）and sodium bicarbonate and then were hydrolyzed with hydrochloric acid. The absorbance of the diluted solution was measured at 346 nm. The other five samples were prepared by the same procedure, except for hydrochlo- ric acid was added before addition of TNBS, when the absorbance was measured as control （A l）. The absorbance after crosslinking：Aaf,~ = ATNBS - A ~. Another 10 samples without any crosslinking were detected with the same procedure to measure the absorbance before crosslinking （Abefore ）. Crosslinking in- dex = （Abloom-Aafter）/Abefore x 100%. （5）Determination of cytotoxicity： two international standard exper- imental methods were adopted in the study accordin

关 键 词：周围神经 胶原 生物相容材料 京尼平 

分 类 号：R318.08[医药卫生—生物医学工程] R329.2[医药卫生—基础医学]