Study on the preparation and physicochemical properties of fish swim bladder membrane_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Lifeng ¹ , GUO Chuan ¹ , SHEN Qiaoqiao ² ,  KONG Qingquan ¹ , WU Jinrong ² , YANG Jin ¹ , WANG Yu ¹ , WU Hao ¹ , PENG Zhiyu ¹ , YAN Yuqing ¹

1. Department of Orthopedics, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China;
2. College of Polymer Science and Engineering, Sichuan University, Chengdu Sichuan, 610065, P.R.China;

Corresponding author：

KONG Qingquan, Email: kqqspine@126.com

Keywords：

Fish swim bladder membrane; crosslinking; physical and chemical properties; dura mater repair

DOI：

10.7507/1002-1892.201809100

Video：

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Objective To manufacture fish swim bladder membrane material by crosslinking techniques, and to explore its physical and chemical properties and cytotoxicity. Methods After decellularization, the swim bladders were randomly divided into two groups. The swim bladders were treated with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) crosslinking method, surface hole making, and freeze-drying in crosslinking group, and only surface hole making and freeze-drying in non-crosslinking group. The physical and chemical properties of the materials were observed, including microstructure by scanning electron microscopy (SEM), mechanical properties (tensile strength and breaking elongation) by universal tensile machine, hydrophilicity by contact angle measuring instrument, porosity by ethanol infiltration method, degradation performance in vitro and thermal stability test, and the components of materials by infrared spectrum analysis. Mouse fibroblasts (L929) were cultured with the extracts of two groups of materials in order to determine the cytotoxicity of materials by using cell counting kit 8 (CCK-8) method. Results The porous structure and rough surface of materials were observed by SEM. Compared with the non-crosslinking group, the tensile stress of the crosslinking group was higher, the breaking elongation was lower, and the porosity increased, showing significant differences (P<0.05). There was no significant difference in contact angle between the two groups (P>0.05). The degradation was faster within the first 7 days and then tended to be smooth in the two groups. But the degradation rates of crosslinking group were significantly lower than those of non-crosslinking group (P<0.05). Differential scanning calorimeter showed that the denaturation temperature of the crosslinking group was (75.2±1.3)℃, which was significantly higher than that of the non-crosslinking group [(68.5±0.4)℃] (t=4.586, P=0.002). Compared with the non-crosslinking group, the crosslinking group produced new C=O bond and N-H bond, and no other new groups were introduced into the cross-linking group. CCK-8 method showed that the absorbance values of the crosslinking group and the non-crosslinking group were not significant when compared with the positive control group (P>0.05). Conclusion The fish swim bladder membrane obtained by crosslinking treatment with EDC/NHS method has good physical and chemical properties, no cytotoxicity, and is expected to be used as a dura mater repair material.

Citation： ZHANG Lifeng, GUO Chuan, SHEN Qiaoqiao, KONG Qingquan, WU Jinrong, YANG Jin, WANG Yu, WU Hao, PENG Zhiyu, YAN Yuqing. Study on the preparation and physicochemical properties of fish swim bladder membrane. Chinese Journal of Reparative and Reconstructive Surgery, 2019, 33(4): 486-491. doi: 10.7507/1002-1892.201809100 Copy

1.	Sairyo K, Matsuura T, Higashino K, et al. Surgery related complications in percutaneous endoscopic lumbar discectomy under local anesthesia. J Med Invest, 2014, 61(3-4): 264-269.
2.	Li H, Dai LY. A systematic review of complications in cervical spine surgery for ossification of the posterior longitudinal ligament. Spine J, 2011, 11(11): 1049-1057.
3.	Boon ME, Ruijgrok JM, Vardaxis MJ. Collagen implants remain supple not allowing fibroblast ingrowth. Biomaterials, 1995, 16(14): 1089-1093.
4.	Berthod F, Hayek D, Damour O, et al. Collagen synthesis by fibroblasts cultured within a collagen sponge. Biomaterials, 1993, 14(10): 749-754.
5.	Schick B, Wolf G, Romeike BF, et al. Dural cell culture. A new approach to study duraplasty. Cells Tissues Organs, 2003, 173(3): 129-137.
6.	Meng F, Modo M, Badylak SF. Biologic scaffold for CNS repair. Regen Med, 2014, 9(3): 367-383.
7.	Sandoval-Sánchez JH, Ramos-Zúñiga R, de Anda SL, et al. A new bilayer chitosan scaffolding as a dural substitute: experimental evaluation. World Neurosurgery, 2012, 77(3-4): 577-582.
8.	Narotam PK, José S, Nathoo N, et al. Collagen matrix (DuraGen) in dural repair: analysis of a new modified technique. Spine (Phila Pa 1976), 2004, 29(24): 2861-2869.
9.	Yamada S, Aiba T, Endo Y, et al. Creutzfeldt-Jakob disease transmitted by a cadaveric dura mater graft. Neurosurgery, 1994, 34(4): 740-744.
10.	Kumar V, Kumar N, Gangwar AK, et al. Comparative histologic and immunologic evaluation of 1,4-butanediol diglycidyl ether crosslinked versus noncrosslinked acellular swim bladder matrix for healing of full-thickness skin wounds in rabbits. J Surg Res, 2015, 197(2): 436-446.
11.	Matsumoto R, Uemura T, Xu Z, et al. Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells. J Biomed Mater Res A, 2015, 103(8): 2531-2539.
12.	Mahesh R, Sreenu M, Kishore PVS, et al. Acellular matrix of swim bladder for cervical oesophagoplasty in rabbits. Trends in Biomaterials and Artificial Organs, 2013, 27(4): 146.
13.	陈泓弛, 位晓娟, 张长青, 等. 鱼胶原蛋白作为新型生物医用材料的研究进展. 中国修复重建外科杂志, 2018, 32(9): 1227-1230.
14.	Pawelec K, Best SM, Cameron RE. Collagen: a network for regenerative medicine. J Mater Chem B Mater Biol Med, 2016, 4(40): 6484-6496.
15.	Dufrane D, Cornu O, Delloye C, et al. Physical and chemical processing for a human dura mater substitute. Biomaterials, 2002, 23(14): 2979-2988.
16.	张更申, 张庆俊, 孙国柱, 等. 应用鲤鱼鳔进行家兔硬脑膜修补术实验研究. 河北医科大学学报, 2000, 21(6): 337-340.
17.	Zerris VA, James KS, Roberts JB, et al. Repair of the dura mater with processed collagen devices. J Biomed Mater Res Part B Appl Biomater, 2007, 83(2): 580-588.
18.	Sacks MS, Jimenez H, Otaño-Lata SE, et al. Local mechanical anisotropy in human cranial dura mater allografts. J Biomech Eng, 1998, 120(4): 541-544.
19.	Li Q, Mu L, Zhang F, et al. A novel fish collagen scaffold as dural substitute. Mater Sci Eng C Mater Biol Appl, 2017, 80: 346-351.
20.	Chen G, Sato T, Ohgushi H, et al. Culturing of skin fibroblasts in a thin PLGA-collagen hybrid mesh. Biomaterials, 2005, 26(15): 2559-2566.

1. Sairyo K, Matsuura T, Higashino K, et al. Surgery related complications in percutaneous endoscopic lumbar discectomy under local anesthesia. J Med Invest, 2014, 61(3-4): 264-269.
2. Li H, Dai LY. A systematic review of complications in cervical spine surgery for ossification of the posterior longitudinal ligament. Spine J, 2011, 11(11): 1049-1057.
3. Boon ME, Ruijgrok JM, Vardaxis MJ. Collagen implants remain supple not allowing fibroblast ingrowth. Biomaterials, 1995, 16(14): 1089-1093.
4. Berthod F, Hayek D, Damour O, et al. Collagen synthesis by fibroblasts cultured within a collagen sponge. Biomaterials, 1993, 14(10): 749-754.
5. Schick B, Wolf G, Romeike BF, et al. Dural cell culture. A new approach to study duraplasty. Cells Tissues Organs, 2003, 173(3): 129-137.
6. Meng F, Modo M, Badylak SF. Biologic scaffold for CNS repair. Regen Med, 2014, 9(3): 367-383.
7. Sandoval-Sánchez JH, Ramos-Zúñiga R, de Anda SL, et al. A new bilayer chitosan scaffolding as a dural substitute: experimental evaluation. World Neurosurgery, 2012, 77(3-4): 577-582.
8. Narotam PK, José S, Nathoo N, et al. Collagen matrix (DuraGen) in dural repair: analysis of a new modified technique. Spine (Phila Pa 1976), 2004, 29(24): 2861-2869.
9. Yamada S, Aiba T, Endo Y, et al. Creutzfeldt-Jakob disease transmitted by a cadaveric dura mater graft. Neurosurgery, 1994, 34(4): 740-744.
10. Kumar V, Kumar N, Gangwar AK, et al. Comparative histologic and immunologic evaluation of 1,4-butanediol diglycidyl ether crosslinked versus noncrosslinked acellular swim bladder matrix for healing of full-thickness skin wounds in rabbits. J Surg Res, 2015, 197(2): 436-446.
11. Matsumoto R, Uemura T, Xu Z, et al. Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells. J Biomed Mater Res A, 2015, 103(8): 2531-2539.
12. Mahesh R, Sreenu M, Kishore PVS, et al. Acellular matrix of swim bladder for cervical oesophagoplasty in rabbits. Trends in Biomaterials and Artificial Organs, 2013, 27(4): 146.
13. 陈泓弛, 位晓娟, 张长青, 等. 鱼胶原蛋白作为新型生物医用材料的研究进展. 中国修复重建外科杂志, 2018, 32(9): 1227-1230.
14. Pawelec K, Best SM, Cameron RE. Collagen: a network for regenerative medicine. J Mater Chem B Mater Biol Med, 2016, 4(40): 6484-6496.
15. Dufrane D, Cornu O, Delloye C, et al. Physical and chemical processing for a human dura mater substitute. Biomaterials, 2002, 23(14): 2979-2988.
16. 张更申, 张庆俊, 孙国柱, 等. 应用鲤鱼鳔进行家兔硬脑膜修补术实验研究. 河北医科大学学报, 2000, 21(6): 337-340.
17. Zerris VA, James KS, Roberts JB, et al. Repair of the dura mater with processed collagen devices. J Biomed Mater Res Part B Appl Biomater, 2007, 83(2): 580-588.
18. Sacks MS, Jimenez H, Otaño-Lata SE, et al. Local mechanical anisotropy in human cranial dura mater allografts. J Biomech Eng, 1998, 120(4): 541-544.
19. Li Q, Mu L, Zhang F, et al. A novel fish collagen scaffold as dural substitute. Mater Sci Eng C Mater Biol Appl, 2017, 80: 346-351.
20. Chen G, Sato T, Ohgushi H, et al. Culturing of skin fibroblasts in a thin PLGA-collagen hybrid mesh. Biomaterials, 2005, 26(15): 2559-2566.

Chinese Journal of Reparative and Reconstructive Surgery

Study on the preparation and physicochemical properties of fish swim bladder membrane

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