EDC交联离子化胶原支架材料的生物学表征

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (4): 343-348.

EDC交联离子化胶原支架材料的生物学表征

王丽霞¹, 赵寰², 靳淑凤¹, 申静¹, 丁一^2*

1. 南开大学口腔医院天津市口腔医院国际诊疗中心天津 300041;
2. 四川大学华西口腔医院牙周病科)

收稿日期:2014-08-27 出版日期:2015-04-28 发布日期:2016-04-29
通讯作者: 丁一,E-mail:yiding2000@126.com
作者简介:王丽霞(1982~ ),女,河北人,主治医师,硕士,主要从事牙周病的病因及防治研究工作。

Biological Characterization of Ionized Collagen Scaffolds with EDC Cross-linking.

WANG Li-xia, ZHAO Huan, JIN Shu-feng, et al

Stomatological Hospital of Nankai University, Tianjin 300041

Received:2014-08-27 Online:2015-04-28 Published:2016-04-29

摘要/Abstract

摘要： 目的:研究EDC交联对离子化胶原支架材料的理化性能的影响。方法:采用侧链修饰结合碳化二亚胺交联改性技术制备实验使用的离子化胶原材料:天然胶原、甲基化胶原、琥珀酰化胶原,通过扫描电境、差示扫描热计、傅里叶变换红外光谱仪和体外酶降解实验分析对胶原材料交联前后的结构与性能进行研究。结果:侧链修饰改变了胶原的表面电荷性质,碳化二亚胺交联提高了天然胶原、甲基化胶原的孔径、孔隙率(P<0.05)、变性温度、抗酶降解能力(P<0.05),改变了其化学结构;而对琥珀酰化胶原的孔径、孔隙率、化学结构无明显影响,且降低了其变性温度和抗酶降解能力(P<0.05)。结论:EDC交联可显著改善离子化胶原支架材料的物理化学性能,为其在牙周组织再生中的初步应用提供了实验基础。

关键词: 碳化二亚胺, 天然胶原, 甲基化胶原, 琥珀酰化胶原

Abstract: Objective: To investigate the effects of EDC cross-linking on the physicochemical properties of ionized collagen scaffolds. Methods: Ionized collagen scaffolds: native collagen(NC), methylation collagen(MC) and succinylated collagen(SC), were fabricated with modification of side chain and 1-ethyl-3-(3- dimethyl-aminopropyl) carbodiimide(EDC)cross-linking. Scaffolds were observed and analyzed using scanning electron microscopy(SEM), differential scanning calorimeter(DSC), Fourier transform infrared radiometer(FT-IR) and experiment of enzyme degradation in vitro. And the effects of EDC cross-linking on the physicochemical properties of collagen scaffolds were investigated. Results: The results indicated that modification of side chain had changed the surface charge of collagen, and EDC cross-linking had improved pore size, porosity, denatured temperature(Td) and ability of antienzyme degradation of NC and MC, changed chemical structure of NC and MC; but that had no influences on pore size, porosity, chemical structure of SC, and decreased Td and ability of antienzyme degradation of it. Coclusion: These data indicate that EDC cross-linking may lead to enhanced physicochemical properties of ionized collagen scaffolds. It provided experimental basis of ionized collagen using in periodontal tissue engineering scaffolds.

Key words: Tissue engineering , Carbodiimide , Collagen, Periodontal ligament fibroblasts, Periodontal tissue regeneration

中图分类号:

R781.4

王丽霞, 赵寰, 靳淑凤, 申静, 丁一. EDC交联离子化胶原支架材料的生物学表征[J]. 口腔医学研究, 2015, 31(4): 343-348.

WANG Li-xia, ZHAO Huan, JIN Shu-feng, et al. Biological Characterization of Ionized Collagen Scaffolds with EDC Cross-linking.[J]. Journal of Oral Science Research, 2015, 31(4): 343-348.

参考文献

[1] 杨志明.组织工程基础与临床[M].成都:四川科学技术出版社,2000,14-25
[2] Kozlovsky A, Aboodi G, Moses O, et al. Bio-degradation of a resorbable collagen membrane(Bio-Gide)applied in a double-layer technique in rats [J]. Clin Qral Implants Res,2009,20(10)∶1116-1123
[3] Niu wen-yu, Wang heng, LinXiu-dan, et al. Study on biocompatibility of a new bovine tendon collagen membrane in vitro [J]. Chinese Journal of Stomatological Research,2012,6(2)∶154-160
[4] Lee CK,Koo KT,Kim TI,et al. Biological effects of a porcine- derived collagen membrane on intrabony defects [J]. J Periodontal Implant Sci,2010,40(5)∶232-238
[5] Bottino MC,Thomas VSchidt G, et al. Recent advances in the development of GTR/GBR membranes for periodontal regeneration-A materials perspective [J]. Dental Materials, 2012, 28(7)∶703-721
[6] Lee CH,Singla A,Lee y. Biomedical applications of collagen [J]. Int J Pharm,2001,22∶1-22
[7] Rault I,Freiv V,Herbage D, et al. Evaluation of different chemical methods for cross-linking collagen gel, films and sponges [J]. J Mater in Med,1996,7(2)∶215-221
[8] Park SN, Lee HJ, Lee KH, et al. Biological characterization of EDC-crosslinked collagen-hyaluronic acid matrix in dermal tissue restoration [J]. Biomaterials,2003,24∶1631-1641
[9] Pieper JS, Oosterhof A, Dijkstra PJ, et al. Preparaion and characterization of porous cross-linking collagenous matrices containing bioavailable chondroitin sulphate [J]. Biomaterials, 1999,20(9)∶847-858
[10] Sivakumar P,Suguna L,Chandrakasan G. Comparative Biochemistry and Physiology Part B, 2000, 125∶555-562
[11] 路易斯,C金奎拉等著,浓宝熔等译.基础组织学[M].山东科学技术出版社,1982∶85-109
[12] 蔡玉文主编.组织学与胚胎学[M].中国中医药出版社,2003∶1
[13] Gerard C,Catuogno C,Amargier-Huin C,et al. The effect of alginate,hyaluronate and hyaluronate derivatives biomaterials on synthesis of non-articular chondrocyte extracellular matrix [J]. J Mater Sci Mater Med, 2005, 16(6)∶541-551
[14] Lisignoli G, Cristino S, Piacentini A, et al. Celular and molecular events during chondrogenesis of human mesenchymal stromal cells grown in a three-dimensional hyaluronan based scaffold [J]. J Biomaterials, 2005, 26(28)∶5677-5686
[15] Teparat T, Solt CW, Clamanl J, et al. Clinical comparison of bioabsothable Barriers with non-resorbable barriers in guided tissue regeneration in the treatment of human intrabony defects [J]. J Periodontol, 1998, 69(6)∶632-641
[16] Thomal DS, Villar CC, Cochran DL, et al. Tissue integration of collagen-based matrices: an experimental study in mice [J]. Clinical Oral Iplants Research,2012,12(23)∶1333-1339
[17] Olde Damink L HH , Dijkst ra PJ , Van Luyn MJA , et al. In vitro degradation of dermal sheep collagen cross-linked using a water-soluble carbodiimide [J]. Biomaterials, 1996, 17(7)∶679-684
[18] Hafemann B, Ghofrani K, Gattner H, et al. Cross-linking by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) of a collagen/ elastin membrane meant to be used as a dermal substitute: effects on physical, biochemical and biological features in vitro [J]. Materials Science-Materials in Medicine, 2001, 12(5)∶437-446
[19] 熊卓,孙磊.骨组织工程聚左旋乳酸多孔框架快速成形研究[J].清华大学学报,自然科学版,2002,42(2)∶157-160
[20] Wake MC, Patrik CW, Mikos AG. Pore morphology affects on the fibro vascular tissue growth in porous polymer substrates [J]. Cell Transplant, 1994, 3(4)∶339-343
[21] M.J.B.Wissink, R. Beernink, J.S. Pieper, et al. Binding and release of basic fibroblast growth factor from heparinized collagen matrices [J]. Biomaterials,2001,22∶2291-2299
[22] Tanaka Y, Kubotal A, Yokokural S, et al. Optical mechanical refinement of human amniotic membrane by dehydration and cross-linking [J]. Journal of Tissue Engineering and Regenerative Medicine,2012,9(6)∶731-737
[23] Cholasa RH, Hsu HP, Spectorb M. The reparative response to cross-linked collagen-based scaffolds in a rat spinal cord gap model [J]. Biomaterials, 2012, 33(7)∶2050-2059