Preparation and biocompatibility of nano polypyrrole/chitin composite membrane_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

 JIAO Haishan ¹ , CAO Ping ¹ , CHEN Ying ² , SONG Yuening ¹ , LI Dongyin ¹ , WANG Xiaodong ²

1. School of Clinical Medicine, Suzhou Vocational Health College, Suzhou Jiangsu, 215009, P.R.China;
2. Department of Histology and Embryology, Medical School of Nantong University, Nantong Jiangsu, 226001, P.R.China;

Corresponding author：

JIAO Haishan, Email: jiaohaishan@szhct.edu.cn

Keywords：

Nanometer; polypyrrole; chitosan; chitin; acetylation; biocompatibility

DOI：

10.7507/1002-1892.201802031

Video：

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Objective To prepare nano polypyrrole (PPy)/chitin composite membrane and observe their biocompatibility. Methods The nano PPy was synthesized by microemulsion polymerization, blended with chitosan and then formed membranes. The membranes were then modified by acetylation to get the experimental membranes (nano PPy/chitin composite membranes, group A). The chitosan membranes (group B) and chitin ones (group C) modified by acetylation acted as control. Scanning electron microscopy and FT-IR spectra were used to identify the nano PPy and the membranes of each group. And the conductivity of membranes of each group was measured. Schwann cells were co-cultured in vitro with each group membranes to observe the biocompatibility by inverted microscope observing, living cell staining, cell counting, and immunofluorescence staining. The lysozyme solution was used to evaluate the degradation of the membranes in vitro. Results The FT-IR spectra showed that the characteristic vibrational absorption peaks of C=C from nano PPy appeared at 1 543.4 cm^–1 and 1 458.4 cm^–1. Scanning electron microscopy observation revealed that the size of nano PPy particles was about 100-200 nm. The nano PPy particles were synthesized. It was successful to turn chitosan to chitin by the acetylation, which was investigated by FT-IR analysis of membranes in groups A and C. The characteristic peaks of the amide Ⅱ band around 1 562 cm^–1 appeared after acetylated modification. Conductivity test showed that the conductivity of membranes in group A was about (1.259 2±0.005 7)×10^–3 S/cm, while the conductivity of the membranes in groups B and C was not detected. The nano PPy particles uniformly distributed on the surface of membranes in group A were observed by scanning electron microscope; the membranes in control groups were smooth. As a result, the nano PPy/chitin composite membranes with electrical conductivity were obtained. The cultured Schwann cells were found to survive with good function by fluorescein diacetate live cell staining, soluble protein-100 immunofluorescence staining, and inverted microscope observing. The cell counting showed that the proliferation of Schwann cells after 2 days and 4 days of group A was more than that of the two control groups, and the differences were significant (P<0.05). It indicated that the nano PPy/chitin composite membranes had better ability of adhesion and proliferation than those of chitosan and chitin membranes. The degradation of membranesin vitro showed that the degradation rates of membranes in groups A and C were significantly higher than those in group B at all time points (P<0.05). In a word, the degradation performance of the membranes modified by acetylation was better than that of chitosan membranes under the same condition. Conclusion The nano PPy and chitosan can be blended and modified by acetylation successfully. Nano PPy/chitin composite membranes had electrical conductivity, degradability, and good biocompatibility in vitro.

Citation： JIAO Haishan, CAO Ping, CHEN Ying, SONG Yuening, LI Dongyin, WANG Xiaodong. Preparation and biocompatibility of nano polypyrrole/chitin composite membrane. Chinese Journal of Reparative and Reconstructive Surgery, 2018, 32(8): 1081-1087. doi: 10.7507/1002-1892.201802031 Copy

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2.	Wang Y, Zhao Y, Sun C, et al. Chitosan degradation products promote nerve regeneration by stimulating Schwann cell proliferation via miR-27a/FOXO1 axis. Mol Neurobiol, 2016, 53(1): 28-39.
3.	Gu J, Hu W, Deng A, et al. Surgical repair of a 30mm long human median nerve defect in the distal forearm by implantation of a chitosan-PGA nerve guidance conduit. J Tissue Eng Regen Med, 2012, 6(2): 163-168.
4.	Jiao H, Yao J, Yang Y, et al. Chitosan/polyglycolic acid nerve grafts for axon regeneration from prolonged axotomized neurons to chronically denervated segments. Biomaterials, 2009, 30(28): 5004-5018.
5.	焦海山, 姚健, 任艳玲, 等. 甲壳素神经导管修复大鼠坐骨神经 10 mm 缺损的实验研究. 中国生物医学工程学报, 2008, 27(4): 597-602.
6.	陈颖, 任艳玲, 李奕, 等. 壳聚糖乙酰化甲壳素材料与许旺细胞的体外相容性. 中国组织工程研究与临床康复, 2008, 12(32): 6230-6234.
7.	Nguyen HT, Sapp S, Wei C, et al. Electric field stimulation through a biodegradable polypyrrole-co-polycaprolactone substrate enhances neural cell growth. J Biomed Mater Res A, 2014, 102(8): 2554-2564.
8.	Balint R, Cassidy NJ, Cartmell SH. Conductive polymers: towards a smart biomaterial for tissue engineering. Acta Biomater, 2014, 10(6): 2341-2353.
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11.	Guimard NK, Sessler JL, Schmidt CE. Towards a biocompatible, biodegradable copolymer incorporating electroactive oligothiophene units. Macromolecules, 2009, 42(2): 502-511.
12.	Wan Y, Wu H, Wen D. Porous-conductive chitosan scaffolds for tissue engineering. I. Preparation and characterization. Macromol Biosci, 2004, 4(9): 882-890.
13.	Yan F, Xue G, Zhou M. Preparation of electrically conducting polypyrrole in oil/water microemulsion. J Appl Polym Sci, 2000, 77: 135-140.
14.	董炎明, 许聪义, 汪剑炜, 等. 红外光谱法测定 N-酰化壳聚糖的取代度. 中国科学 (B 辑), 2001, 31(2): 153-160.
15.	Poletti Papi MA, Caetano FR, Bergamini MF, et al. Facile synthesis of a silver nanoparticles/polypyrrole nanocomposite for non-enzymatic glucose determination. Mater Sci Eng C Mater Biol Appl, 2017, 75: 88-94.
16.	王爱勤. 甲壳素化学. 北京: 科学出版社, 2008: 226-227.
17.	Freier T, Koh HS, Kazazian K, et al. Controlling cell adhesion and degradation of chitosan films by N-acetylation. Biomaterials, 2005, 26(29): 5872-5878.
18.	Foster LJ, Ho S, Hook J, et al. Chitosan as a biomaterial: influence of degree of deacetylation on its physiochemical, material and biological properties. PLoS One, 2015, 10(8): e0135153.
19.	Jang LK, Kim S, Seo J, et al. Facile and controllable electrochemical fabrication of cell-adhesive polypyrrole electrodes using pyrrole-RGD peptides. Biofabrication, 2017, 9(4): 045007.
20.	Wang X, Gu X, Yuan C, et al. Evaluation of biocompatibility of polypyrrole in vitro and in vivo. J Biomed Mater Res A, 2004, 68(3): 411-422.
21.	Fahlgren A, Bratengeier C, Gelmi A, et al. Biocompatibility of polypyrrole with human primary osteoblasts and the effect of dopants. PLoS One, 2015, 10(7): e0134023.
22.	Wan Y, Yu A, Wu H, et al. Porous-conductive chitosan scaffolds for tissue engineering Ⅱ. In vitro and in vivo degradation. J Mater Sci Mater Med, 2005, 16(11): 1017-1028.
23.	熊敏剑, 李晓峰, 闵燕, 等. 不同脱乙酰度壳聚糖支架制备及降解性能评价. 生物医学工程学杂志, 2012, 29(1): 107-111.
24.	Yang YM, Hu W, Wang XD, et al. The controlling biodegradation of chitosan fibers by N-acetylation in vitro and in vivo. J Mater Sci Mater Med, 2007, 18(11): 2117-2121.

1. Wang XD, Hu W, Cao Y, et al. Dog sciatic nerve regeneration across a 30-mm defect bridged by a chitosan/PGA artificial nerve graft. Brain, 2005, 128(8): 1897-1910.
2. Wang Y, Zhao Y, Sun C, et al. Chitosan degradation products promote nerve regeneration by stimulating Schwann cell proliferation via miR-27a/FOXO1 axis. Mol Neurobiol, 2016, 53(1): 28-39.
3. Gu J, Hu W, Deng A, et al. Surgical repair of a 30mm long human median nerve defect in the distal forearm by implantation of a chitosan-PGA nerve guidance conduit. J Tissue Eng Regen Med, 2012, 6(2): 163-168.
4. Jiao H, Yao J, Yang Y, et al. Chitosan/polyglycolic acid nerve grafts for axon regeneration from prolonged axotomized neurons to chronically denervated segments. Biomaterials, 2009, 30(28): 5004-5018.
5. 焦海山, 姚健, 任艳玲, 等. 甲壳素神经导管修复大鼠坐骨神经 10 mm 缺损的实验研究. 中国生物医学工程学报, 2008, 27(4): 597-602.
6. 陈颖, 任艳玲, 李奕, 等. 壳聚糖乙酰化甲壳素材料与许旺细胞的体外相容性. 中国组织工程研究与临床康复, 2008, 12(32): 6230-6234.
7. Nguyen HT, Sapp S, Wei C, et al. Electric field stimulation through a biodegradable polypyrrole-co-polycaprolactone substrate enhances neural cell growth. J Biomed Mater Res A, 2014, 102(8): 2554-2564.
8. Balint R, Cassidy NJ, Cartmell SH. Conductive polymers: towards a smart biomaterial for tissue engineering. Acta Biomater, 2014, 10(6): 2341-2353.
9. Guo B, Glavas L, Albertsson AC. Biodegradable and electrically conducting polymers for biomedical applications. Progress in Polymer Science, 2013, 38(9): 1263-1286.
10. Shi G, Rouabhia M, Wang Z, et al. A novel electrically conductive and biodegradable composite made of polypyrrole nanoparticles and polylactide. Biomaterials, 2004, 25(13): 2477-2488.
11. Guimard NK, Sessler JL, Schmidt CE. Towards a biocompatible, biodegradable copolymer incorporating electroactive oligothiophene units. Macromolecules, 2009, 42(2): 502-511.
12. Wan Y, Wu H, Wen D. Porous-conductive chitosan scaffolds for tissue engineering. I. Preparation and characterization. Macromol Biosci, 2004, 4(9): 882-890.
13. Yan F, Xue G, Zhou M. Preparation of electrically conducting polypyrrole in oil/water microemulsion. J Appl Polym Sci, 2000, 77: 135-140.
14. 董炎明, 许聪义, 汪剑炜, 等. 红外光谱法测定 N-酰化壳聚糖的取代度. 中国科学 (B 辑), 2001, 31(2): 153-160.
15. Poletti Papi MA, Caetano FR, Bergamini MF, et al. Facile synthesis of a silver nanoparticles/polypyrrole nanocomposite for non-enzymatic glucose determination. Mater Sci Eng C Mater Biol Appl, 2017, 75: 88-94.
16. 王爱勤. 甲壳素化学. 北京: 科学出版社, 2008: 226-227.
17. Freier T, Koh HS, Kazazian K, et al. Controlling cell adhesion and degradation of chitosan films by N-acetylation. Biomaterials, 2005, 26(29): 5872-5878.
18. Foster LJ, Ho S, Hook J, et al. Chitosan as a biomaterial: influence of degree of deacetylation on its physiochemical, material and biological properties. PLoS One, 2015, 10(8): e0135153.
19. Jang LK, Kim S, Seo J, et al. Facile and controllable electrochemical fabrication of cell-adhesive polypyrrole electrodes using pyrrole-RGD peptides. Biofabrication, 2017, 9(4): 045007.
20. Wang X, Gu X, Yuan C, et al. Evaluation of biocompatibility of polypyrrole in vitro and in vivo. J Biomed Mater Res A, 2004, 68(3): 411-422.
21. Fahlgren A, Bratengeier C, Gelmi A, et al. Biocompatibility of polypyrrole with human primary osteoblasts and the effect of dopants. PLoS One, 2015, 10(7): e0134023.
22. Wan Y, Yu A, Wu H, et al. Porous-conductive chitosan scaffolds for tissue engineering Ⅱ. In vitro and in vivo degradation. J Mater Sci Mater Med, 2005, 16(11): 1017-1028.
23. 熊敏剑, 李晓峰, 闵燕, 等. 不同脱乙酰度壳聚糖支架制备及降解性能评价. 生物医学工程学杂志, 2012, 29(1): 107-111.
24. Yang YM, Hu W, Wang XD, et al. The controlling biodegradation of chitosan fibers by N-acetylation in vitro and in vivo. J Mater Sci Mater Med, 2007, 18(11): 2117-2121.

Chinese Journal of Reparative and Reconstructive Surgery

Preparation and biocompatibility of nano polypyrrole/chitin composite membrane

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