丝素蛋白/左旋聚乳酸支架材料与软骨细胞体外细胞相容性研究

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (3): 221-225.

丝素蛋白/左旋聚乳酸支架材料与软骨细胞体外细胞相容性研究

孙莹¹, 李正强¹, 刘炜炜¹, 郑璐², 韩冰^1*

1. 吉林大学口腔医院口腔颌面外科吉林长春 130021;
2. 吉林大学化学学院应用化学

收稿日期:2014-08-07 出版日期:2015-03-28 发布日期:2016-04-29
通讯作者: 韩冰,电话:0431-85579316
作者简介:孙莹(1987~ ),女,山东济南人,硕士在读,主要从事口腔颌面外科肿瘤学及损伤的研究。
基金资助:
吉林省科技厅科研基金资助课题(3D512K513431)

Cytocompatibility Study of Silk Fibroin/poly-L-lactic Acid Scaffold and Chondrocytes in Vitro.

SUN Ying, LI Zheng-qiang, LIU Wei-wei, et al.

Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Jilin University, Changchun 130021

Received:2014-08-07 Online:2015-03-28 Published:2016-04-29

摘要/Abstract

摘要： 目的:观察静电纺丝支架材料丝素蛋白/左旋聚乳酸(SF/PLLA)的体外细胞相容性,探索其作为软骨组织工程支架材料的可行性,为进一步的体内及动物实验奠定基础。方法:将兔膝关节软骨细胞与丝素蛋白/左旋聚乳酸(SF/PLLA)支架材料复合培养,在第3、7、14天分别作HE染色和阿利新蓝+核固红染色,扫描电镜检验细胞黏着情况,MTT试验检测细胞在支架上的增殖情况。结果:细胞在支架上可以获得良好的粘附,细胞增殖良好,无细胞表型的变化。结论:丝素蛋白/左旋聚乳酸(SF/PLLA)具有良好的细胞相容性,可作为软骨组织工程支架材料。

关键词: 丝素蛋白/左旋聚乳酸(SF/PLLA), 组织工程, 软骨

Abstract: Objective: To lay the foundation for further body and animal experiment, we observe the cytocompatibility on electrostatic spinning scaffold of silk fibroin/poly-L-lactic acid (SF/PLLA) in vitro and explore the feasibility in cartilage tissue engineering scaffolds. Methods: The rabbit knee joint chondrocyte were co-cultured with silk fibroin/poly-L-lactic acid (SF/PLLA) scaffold. Hematoxylin-eosin staining and Alcian blue- nuclear fast red staining were tested on day 3, 7 and 14 respectively. Cell adhesion was tested by scanning electron microscopy (SEM). Cell proliferation was determined by MTT test. Results: Adhesion, proliferation, and growth of chondrocyte derived from rabbit knee joint chondrocytes were great on the surface of silk fibroin/poly-L-lactic acid (SF/PLLA). The chondrocytes cultured in vitro maintained the specific chondrocytes phenotype on the scaffold. Conclusion: Silk fibroin/poly-L-lactic acid (SF/PLLA) scaffold exhibits excellent cytocompatibility, and can be used for cartilage tissue engineering.

Key words: Silk fibroin/poly-L-lactic acid (SF/PLLA), Tissue engineering, Cartilage

中图分类号:

R782.2

孙莹, 李正强, 刘炜炜, 郑璐, 韩冰. 丝素蛋白/左旋聚乳酸支架材料与软骨细胞体外细胞相容性研究[J]. 口腔医学研究, 2015, 31(3): 221-225.

SUN Ying, LI Zheng-qiang, LIU Wei-wei, et al.. Cytocompatibility Study of Silk Fibroin/poly-L-lactic Acid Scaffold and Chondrocytes in Vitro.[J]. Journal of Oral Science Research, 2015, 31(3): 221-225.

参考文献

[1] Formhals A. Process and apparatus fob pbepabing: U.S. Patent 1,975,504[P]. 1934-10-2
[2] Ma Z, Kotaki M, Inai R, et al. Potential of nanofiber matrix as tissue-engineering scaffolds [J]. Tissue engineering, 2005, 11(1-2)∶101-109
[3] Altman G H, Diaz F, Jakuba C, et al. Silk-based biomaterials [J]. Biomaterials, 2003, 24(3)∶401-416
[4] Vepari C, Kaplan D L. Silk as a biomaterial [J]. Progress in polymer science, 2007, 32(8)∶991-1007
[5] 张丽斯. 丝素蛋白/聚乳酸复合组织工程纤维支架制备及性能研究[D]. 吉林大学,2012
[6] 张晓燕, 李正强, 孙莹, 等. 丝素蛋白/左旋聚乳酸复合组织工程纳米材料的生物相容性及安全性评价[J].吉林大学学报(医学版),40(03)∶578-582
[7] Li W J, Laurencin C T, Caterson E J, et al. Electrospun nanofibrous structure: a novel scaffold for tissue engineering [J]. Journal of biomedical materials research, 2002, 60(4)∶613-621
[8] Xiao W, He J, Nichol J W, et al. Synthesis and characterization of photocrosslinkable gelatin and silk fibroin interpenetrating polymer network hydrogels [J]. Acta biomaterialia, 2011, 7(6)∶2384-2393
[9] Stoppato M, Stevens H Y, Carletti E, et al. Effects of silk fibroin fiber incorporation on mechanical properties, endothelial cell colonization and vascularization of PDLLA scaffolds [J]. Biomaterials, 2013, 34(19)∶4573-4581
[10] Schnabel M, Marlovits S, Eckhoff G, et al. Dedifferentiation-associated changes in morphology and gene expression in primary human articular chondrocytes in cell culture [J]. Osteoarthritis and Cartilage, 2002, 10(1)∶62-70
[11] Fan Z, Zhang F, Liu T, et al. Effect of hyaluronan molecular weight on structure and biocompatibility of silk fibroin/hyaluronan scaffolds [J]. International journal of biological macromolecules, 2014, 65∶516-523
[12] Amornsudthiwat P, Mongkolnavin R, Kanokpanont S, et al. Improvement of early cell adhesion on Thai silk fibroin surface by low energy plasma [J]. Colloids and Surfaces B: Biointerfaces, 2013, 111∶579-586
[13] Holy C E, Shoichet M S, Davies J E. Engineering three-dimensional bone tissue in vitro using biodegradable scaffolds: investigating initial cell-seeding density and culture period [J]. Journal of biomedical materials research, 2000, 51(3)∶376-382
[14] 苗宗宁,潘宇红,祝建中,等.丝素蛋白支架材料复合骨髓间充质干细胞构建组织工程化软骨[J].中国组织工程研究与临床康复,2008,12(27)∶5243-5247
[15] Deng J, She R, Huang W, et al. A silk fibroin/chitosan scaffold in combination with bone marrow-derived mesenchymal stem cells to repair cartilage defects in the rabbit knee [J]. Journal of Materials Science: Materials in Medicine, 2013, 24(8)∶2037-2046
[16] Teuschl A H, Neutsch L, Monforte X, et al. Enhanced cell adhesion on silk fibroin via lectin surface modification [J]. Acta biomaterialia, 2014, 10(6)∶2506-2517