STUDY ON MODIFICATION OF BIOMATERIALS OF ACELLULAR BOVINE PERICARDIUM WITH DIFFERENT CROSSLINKING REAGENTS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

 XIAOHongtao , TIANShemin , ZHAXinjian , WEIYing , HUANGHongjun , LIYun , YANGHuanna , XIAChengde , NIUXihua

Department of Burns, No. 1 People's Hospital of Zhengzhou, Zhengzhou Henan, 450004, P. R. China;

Corresponding?author：

XIAOHongtao, Email: htxiao917@126.com

Keywords：

Acellular bovine pericardium; Crosslinking reagent; Genipin; 1-ethyl-3-(3-dinethylami-nopropyl)carbodimide; Biological property

DOI：

10.7507/1002-1892.20150281

Video：

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Objective To investigate the effects of modification of acellular bovine pericardium with 1-ethyl-3-(3-dinethylami-nopropyl) carbodimide (EDC)/N-hydroxysuccininide (NHS) or genipin and find out the best crosslinking reagent. Methods The cellular components of the bovine pericardiums were removed. The effects of decellularization were tested by HE staining. The acellular bovine pericardiums were crosslinked with EDC/NHS (EDC/NHS group) or genipin (genipin group). The properties of the crosslinked acellular matrix were evaluated by scanning electron microscope (SEM), matrix thickness, crosslinking index, mechanical property, denaturation temperature, enzymatic degradation, and cytotoxicity test before and after the crosslinking. Acellular bovine pericardium (ABP group) or normal bovine pericardium (control group) were harvested as controls. Results SEM showed that collagen fibers were reticulated in bovine pericardial tissues after crosslinked by EDC/NHS or genipin, and relative aperture of the collagen fiber was from 10 to 20 μm. The thickness and denaturation temperature of the scaffolds were increased significantly after crosslinking with EDC/NHS or genipin (P<0.05), while there was no significant difference between EDC/NHS group and genipin group (P>0.05). The difference had no statistical significance in crosslinking index between EDC/NHS group and genipin group (t=0.205, P=0.218). The degradation rate in EDC/NHS group and genipin group was significantly lower than that in ABP group and control group (P<0.05). Elastic modulus and fracture stress in EDC/NHS group and genipin group were significantly lower than those in ABP group (P<0.05), but there was no significant difference among EDC/NHS group, genipin group, and control group (P>0.05). The break elongation in EDC/NHS group and genipin group were significantly increased than those in ABP group and control group (P<0.05). The difference had no statistical significance in stability and mechanical properties between EDC/NHS group and genipin group (P>0.05). Cytotoxicity of genipin crosslinked tissue (grade 1) were much lower than that of EDC/NHS (grade 2) at 5 days. Conclusion Acellular bovine pericardium crosslinked with genipin has better biocompatibility than EDC/NHS.

Citation： XIAOHongtao, TIANShemin, ZHAXinjian, WEIYing, HUANGHongjun, LIYun, YANGHuanna, XIAChengde, NIUXihua. STUDY ON MODIFICATION OF BIOMATERIALS OF ACELLULAR BOVINE PERICARDIUM WITH DIFFERENT CROSSLINKING REAGENTS. Chinese Journal of Reparative and Reconstructive Surgery, 2015, 29(10): 1301-1306. doi: 10.7507/1002-1892.20150281 Copy

1.	Strange G, Brizard C, Karl TR, et al. An evaluation of Admedus' tissue engineering process-treated (ADAPT) bovine pericardium patch (CardioCel) for the repair of cardiac and vascular defects. Expert Rev Med Devices, 2015, 12(2):135-141.
2.	Dorweiler B, Kayser C, Zipp F, et al. Long-term performance of the bovine pericardium patch in conventional carotid endarterectomy. Thorac Cardiovasc Surg, 2015, 63(2):168-174.
3.	Gubitosi A, Docimo G, Parmeggiani D, et al. Acellular bovine pericardium dermal matrix in immediate breast reconstruction after Skin Sparing Mastectomy. Int J Surg, 2014, 12 Suppl 1:S205-208.
4.	Leon-Takahashi AM, Gomez-Pedraza A, Cruz-Rodriguez A, et al. Use of glutaraldehyde-preserved bovine pericardium patch in vascular repair of portal vein in a patient with pancreatic cancer. Indian J Surg, 2013, 75(Suppl 1):247-249.
5.	Barros JA, Filippin-Monteiro FB, de Oliveira EM, et al. Cytotoxicity of PVPAC-treated bovine pericardium:a potential replacement for glutaraldehyde in biological heart valves. J Biomed Mater Res B Appl Biomater, 2014, 102(3):574-582.
6.	Shepherd DV, Shepherd JH, Ghose S, et al. The process of EDCNHS cross-linking of reconstituted collagen fibres increases collagen fibrillar order and alignment. APL Mater, 2015, 3(1):014902.
7.	王旻, 笪琳萃, 謝艷, 等. 京尼平作為交聯劑在天然生物材料改性中的應用. 中國修復重建外科雜志, 2013, 27(5):580-585.
8.	Maizato MJ, Higa OZ, Mathor MB, et al. Glutaraldehyde-treated bovine pericardium:effects of lyophilization on cytotoxicity and residual aldehydes. Artif Organs, 2003, 27(8):692-694.
9.	Chang Y, Tsai CC, Liang HC, et al. In vivo evaluation of cellular and acellular bovine pericardia fixed with a naturally occurring crosslinking agent (genipin). Biomaterials, 2002, 23(12):2447-2457.
10.	董小超. RGD交聯去細胞牛心包構建組織工程心臟瓣膜支架的實驗研究. 西安:第四軍醫大學, 2009.
11.	王迎軍, 楊春蓉, 汪凌云. EDC/NHS交聯對膠原物理化學性能的影響. 華南理工大學學報:自然科學版, 2007, 35(12):66-70, 96.
12.	姚芳蓮, 李學強, 于瀟, 等. 京尼平對殼聚糖及明膠的交聯反應. 天津大學學報, 2007, 40(12):1485-1489.
13.	馬兵, 翟萬銀, 張嘉敏, 等. PC交聯脫細胞牛心包膜的力學性能、穩定性和血液相容性. 華東師范大學學報:自然科學版, 2013, 59(5):61-70, 79.
14.	吳彤, 王瑞明, 黃磊, 等. 蛋白酶K的研究進展. 食品工業科技, 2013, 34(14):380-384.
15.	陳平, 李新華, 邢萬紅. 京尼平交聯的脫細胞牛心包生物支架材料的實驗研究. 中國醫藥導報, 2010, 7(6):28-30.
16.	Brizard CP, Brink J, Horton SB, et al. New engineering treatment of bovine pericardium confers outstanding resistance to calcification in mitral and pulmonary implantations in a juvenile sheep model. J Thorac Cardiovasc Surg, 2014, 148(6):3194-3201.
17.	Scheffel D, Bianchi L, Soares D, et al. Transdentinal cytotoxicity of carbodiimide (EDC) and glutaraldehyde on odontoblast-like cells. Oper Dent, 2015, 40(1):44-54.
18.	田聰, 萬榮欣, 劉欣, 等. 生物交聯劑京尼平交聯牛心包生物支架材料的性能. 中國生物醫學工程學報, 2011, 30(2):281-286.
19.	汪紅, 蘇建, 高陪, 等. 不同pH值對京尼平交聯膠原制備生物材料的影響. 四川大學學報:自然科學版, 2011, 48(2):481-485.
20.	曹崢, 胡蕰玉, 潘緯敏, 等. 交聯溫度對京尼平交聯膠原/殼聚糖組織工程支架的影響. 中國矯形外科雜志, 2009, 17(3):217-220.
21.	Gratzer PF, Lee JM. Control of pH alters the type of cross-linking produced by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) treatment of acellular matrix vascular grafts. J Biomed Mater Res, 2001, 58(2):172-179.
22.	吳秀娟, 謝明均. 京尼平對去細胞組織修飾的研究進展. 中華臨床醫師雜志(電子版), 2014, 8(13):2532-2535.
23.	秦勝男, 陳鴻輝, 楊小紅, 等. 經碳化二胺/N-羥基琥珀酰亞胺交聯的Ⅱ型膠原海綿的制備. 中國矯形外科雜志, 2010, 18(12):1014-1018.

1. Strange G, Brizard C, Karl TR, et al. An evaluation of Admedus' tissue engineering process-treated (ADAPT) bovine pericardium patch (CardioCel) for the repair of cardiac and vascular defects. Expert Rev Med Devices, 2015, 12(2):135-141.
2. Dorweiler B, Kayser C, Zipp F, et al. Long-term performance of the bovine pericardium patch in conventional carotid endarterectomy. Thorac Cardiovasc Surg, 2015, 63(2):168-174.
3. Gubitosi A, Docimo G, Parmeggiani D, et al. Acellular bovine pericardium dermal matrix in immediate breast reconstruction after Skin Sparing Mastectomy. Int J Surg, 2014, 12 Suppl 1:S205-208.
4. Leon-Takahashi AM, Gomez-Pedraza A, Cruz-Rodriguez A, et al. Use of glutaraldehyde-preserved bovine pericardium patch in vascular repair of portal vein in a patient with pancreatic cancer. Indian J Surg, 2013, 75(Suppl 1):247-249.
5. Barros JA, Filippin-Monteiro FB, de Oliveira EM, et al. Cytotoxicity of PVPAC-treated bovine pericardium:a potential replacement for glutaraldehyde in biological heart valves. J Biomed Mater Res B Appl Biomater, 2014, 102(3):574-582.
6. Shepherd DV, Shepherd JH, Ghose S, et al. The process of EDCNHS cross-linking of reconstituted collagen fibres increases collagen fibrillar order and alignment. APL Mater, 2015, 3(1):014902.
7. 王旻, 笪琳萃, 謝艷, 等. 京尼平作為交聯劑在天然生物材料改性中的應用. 中國修復重建外科雜志, 2013, 27(5):580-585.
8. Maizato MJ, Higa OZ, Mathor MB, et al. Glutaraldehyde-treated bovine pericardium:effects of lyophilization on cytotoxicity and residual aldehydes. Artif Organs, 2003, 27(8):692-694.
9. Chang Y, Tsai CC, Liang HC, et al. In vivo evaluation of cellular and acellular bovine pericardia fixed with a naturally occurring crosslinking agent (genipin). Biomaterials, 2002, 23(12):2447-2457.
10. 董小超. RGD交聯去細胞牛心包構建組織工程心臟瓣膜支架的實驗研究. 西安:第四軍醫大學, 2009.
11. 王迎軍, 楊春蓉, 汪凌云. EDC/NHS交聯對膠原物理化學性能的影響. 華南理工大學學報:自然科學版, 2007, 35(12):66-70, 96.
12. 姚芳蓮, 李學強, 于瀟, 等. 京尼平對殼聚糖及明膠的交聯反應. 天津大學學報, 2007, 40(12):1485-1489.
13. 馬兵, 翟萬銀, 張嘉敏, 等. PC交聯脫細胞牛心包膜的力學性能、穩定性和血液相容性. 華東師范大學學報:自然科學版, 2013, 59(5):61-70, 79.
14. 吳彤, 王瑞明, 黃磊, 等. 蛋白酶K的研究進展. 食品工業科技, 2013, 34(14):380-384.
15. 陳平, 李新華, 邢萬紅. 京尼平交聯的脫細胞牛心包生物支架材料的實驗研究. 中國醫藥導報, 2010, 7(6):28-30.
16. Brizard CP, Brink J, Horton SB, et al. New engineering treatment of bovine pericardium confers outstanding resistance to calcification in mitral and pulmonary implantations in a juvenile sheep model. J Thorac Cardiovasc Surg, 2014, 148(6):3194-3201.
17. Scheffel D, Bianchi L, Soares D, et al. Transdentinal cytotoxicity of carbodiimide (EDC) and glutaraldehyde on odontoblast-like cells. Oper Dent, 2015, 40(1):44-54.
18. 田聰, 萬榮欣, 劉欣, 等. 生物交聯劑京尼平交聯牛心包生物支架材料的性能. 中國生物醫學工程學報, 2011, 30(2):281-286.
19. 汪紅, 蘇建, 高陪, 等. 不同pH值對京尼平交聯膠原制備生物材料的影響. 四川大學學報:自然科學版, 2011, 48(2):481-485.
20. 曹崢, 胡蕰玉, 潘緯敏, 等. 交聯溫度對京尼平交聯膠原/殼聚糖組織工程支架的影響. 中國矯形外科雜志, 2009, 17(3):217-220.
21. Gratzer PF, Lee JM. Control of pH alters the type of cross-linking produced by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) treatment of acellular matrix vascular grafts. J Biomed Mater Res, 2001, 58(2):172-179.
22. 吳秀娟, 謝明均. 京尼平對去細胞組織修飾的研究進展. 中華臨床醫師雜志(電子版), 2014, 8(13):2532-2535.
23. 秦勝男, 陳鴻輝, 楊小紅, 等. 經碳化二胺/N-羥基琥珀酰亞胺交聯的Ⅱ型膠原海綿的制備. 中國矯形外科雜志, 2010, 18(12):1014-1018.

Chinese Journal of Reparative and Reconstructive Surgery

STUDY ON MODIFICATION OF BIOMATERIALS OF ACELLULAR BOVINE PERICARDIUM WITH DIFFERENT CROSSLINKING REAGENTS

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