研究了温度梯度法所制再生丝素蛋白膜的溶失率、抗酶能力和动物细胞体外培养特性.以紫外分光光度法检测丝素蛋白膜在水中的溶失率;再用质量体积浓度0.25％和0.5％的胰蛋白酶液分别消化丝素蛋白膜2d,以膜的结构和质量是否发生明显变化为指标,判断膜抗胰蛋白酶的能力;最后在丝素蛋白膜上培养BHK-21细胞和Vero细胞,再用0.25％和0.5％胰蛋白酶液分别消化膜上细胞并进行传代培养,观察细胞形态和生长状况.实验结果表明,温度梯度法直接所制丝素蛋白膜48 h内的溶失率低于0.2％,再经25％甲醇处理后溶失率为0,2d内膜仍保持完整结构且质量无明显变化;BHK-21细胞和Vero细胞在丝素蛋白膜上生长良好,形态正常;0.25％和0.5％的胰蛋白酶液均可在4 min内将膜上两种细胞消化脱落且细胞连续传代培养5代,细胞形态正常.
参考文献
| [1] | Shi P;Goh JC .Release and cellular acceptance of multiple drugs loaded silk fibroin particles.[J].International Journal of Pharmaceutics,2011(2):282-289. |
| [2] | Thanonchat Imsombut;Yaowalak Srisuwan;Prasong Srihanam .Genipin-cross-linked silk fibroin microspheres prepared by the simple water-in-oil emulsion solvent diffusion method[J].Powder Technology: An International Journal on the Science and Technology of Wet and Dry Particulate Systems,2010(3):603-608. |
| [3] | 王学超,李延报,陆春华,许仲梓.水溶性丝素蛋白的制备及其构象转变[J].南京工业大学学报（自然科学版）,2013(02):55-58. |
| [4] | Hyeon Yoon;Eun Young Kim;Hyeongseok Kim .Fabrication of Transparent Silk Fibroin Film for the Regeneration of Corneal Endothelial Cells; Preliminary Study[J].Macromolecular Research,2014(3):297-303. |
| [5] | Andiyappan Muthumanickkam;Sundaramoorthy Subramanian;Mahizhappan Goweri;Winfred Sofi Beaula;Venkataraman Ganesh .Comparative study on eri silk and mulberry silk fibroin scaffolds for biomedical applications[J].Iranian polymer journal,2013(3):143-154. |
| [6] | Kar S;Talukdar S;Pal S et al.Silk gland fibroin from indian muga silkworm Antheraea assama as potential biomaterial[J].J Tissue Eng Regen Med,2013,10(04):201. |
| [7] | Bray L J;George K A;Suzuki S.Fabrication of a corneal-limbal tissue substitute using silk fibroin[M].New York:Humana Press,2013 |
| [8] | 焦宇虹,李明忠.蚕丝丝素蛋白材料交联方法的研究进展[J].现代丝绸科学与技术,2011(06):242-246. |
| [9] | 田莉,陈宇,闵思佳.环氧化合物制备丝素凝胶材料的细胞毒性研究[J].生物医学工程学杂志,2007(06):1309-1313. |
| [10] | Vepari C;Kaplan D L .Silk as a biomaterial[J].Progress in Polymer Science,2007,32(08):991. |
| [11] | David N. Breslauer;Susan J. Muller;Luke P. Lee .Generation of Monodisperse Silk Microspheres Prepared with Microfluidics[J].Biomacromolecules,2010(3):643-647. |
| [12] | David L;Spector M;Robert D.Cells a laboratory manual[M].California:Cold Spring Harbor Laboratory Press,1998 |
| [13] | 王志强,梁锐,陈明清,邓俊,李文亮,陈天星,文政琦,杨军.胰蛋白酶消化法和组织块法原代培养人包皮成纤维细胞的比较[J].生命科学研究,2012(03):242-247. |
| [14] | 胡银英,袁铿,戴志芳,袁芳.胰蛋白酶在原代细胞制备中的灵活应用[J].江西医学院学报,2002(01):43-44. |
| [15] | 毛丽,张岑岑,吴锡龙,卢神州.柞蚕丝素/丙二醇共混膜的制备及性能测试[J].蚕业科学,2011(03):461-465. |
| [16] | Magoshi J;Magoshi Y;Nakamura S et al.Physical properties and structure of silk.Ⅴ.Thermal behavior of silk fibroin in the random-coil conformation[J].JOURNAL OF POLYMER SCIENCE PART B:POLYMER PHYSICS,1977,15(09):1675. |
| [17] | Magoshi J;Magoshi Y;Nakamura S .Physical properties and structure of silk.Ⅷ.Crystallization of amorphous silk fibroin induced by immersion in methanol[J].JOURNAL OF POLYMER SCIENCE PART B:POLYMER PHYSICS,1981,19(01):185. |
| [18] | 徐亚梅 .丝素蛋白材料生物降解行为的研究[D].苏州大学,2011. |
| [19] | Lin CY;Kikuchi N;Hollister SJ .A novel method for biomaterial scaffold internal architecture design to match bone elastic properties with desired porosity[J].Journal of Biomechanics,2004(5):623-636. |
| [20] | 魏凤,管宇,肖跃强,吕素芳,张文通,沈志强.微载体技术在动物细胞培养中的应用进展[J].黑龙江畜牧兽医（上半月）,2013(09):34-36. |
| [21] | Frederic Byette;Frederic Bouchard;Christian Pellerin .Cell-culture compatible silk fibroin scaffolds concomitantly patterned by freezing conditions and salt concentration[J].Polymer bulletin,2011(1):159-175. |
| [22] | Navone S E;Pascucci L;Dossena M et al.Decellularized silk fibroin scaffold primed with adipose mesenchymal stromal cells improves wound healing in diabetic mice[J].Stem Cell Res Ther,2014,5(07):2. |
上一张
下一张
上一张
下一张
计量
- 下载量()
- 访问量()
文章评分
- 您的评分：
-
10%
-
20%
-
30%
-
40%
-
50%