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	In vitro biocompatibility of three chitosan/polycation composite materials for nerve regeneration
	Zheng, Zhenhuan ; Wei, Yujun ; Wang, Gan ; Gong, Yandao ; Zhang, Xiufang
	2010-05-11 ; 2010-05-11
关键词	biocompatibility chitosan nerve regeneration polycation NEURITE OUTGROWTH SCHWANN-CELLS REPAIR STIMULATION SCAFFOLDS CONDUITS Cell Biology Neurosciences
中文摘要	BACKGROUND: It has been reported that chitosan nerve conduits could support axon elongation and improve relevant function during in vivo nerve regeneration. OBJECTIVE: To investigate in vitro biocompatibility of three novel, chitosan/polycation composite materials for nerve regeneration ill cultured mouse Schwann cells and PC12 cells. DESIGN, TIME AND SETTING: The observational, control experiments for nerve tissue engineering were performed at the Department of Biological Sciences and Biotechnology of Tsinghua University front August 2007 to January 2008. MATERIALS: Mouse Schwann cells were isolated from the sciatic nerve of 5-7-day-old BALB/C mice. PC12 cells were purchased from the American Type Culture Collection (ATCC, USA). Chitosan was purchased from Tsingdao Haisheng Co., China. Poly-L-lysine hydrochloride (PLL), polyethyleneimine (PEI), poly-L-ornithine hydrobromide (POR), and S-100 antibody was purchased from Sigma Chemical Co., USA. Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Chemical Co., Japan. METHODS: Three chitosan/polycation composite materials for nerve regeneration (PLL-0.25, PEI-0.25, and POR-0.25) were produced by blending chitosan with 0.25% (w/w) poly-L-lysine, polyethyleneimine, and poly-L-ornithine. Pure chitosan was utilized as the control. After 3 days Of Culture, the morphology of mouse Schwann and PC12 cells cultured oil all substrates was observed with all inverted phase contrast microscope. Mouse Schwann cells were stained by immunofluorescence labeling S-100 protein and nuclei, followed by identification with a confocal laser-scanning microscope. The amount of proliferating Mouse Schwann and PC12 cells was determined by CCK-8 after 1, 3, and 5 days in culture. The level of PC12 cell differentiation oil all Substrates was assessed by measuring neurite length at 1, 3, and 5 days after seeding. MAIN OUTCOME MEASURES: Morphology and amount of proliferation of mouse Schwann cells and PC12 Cells Cultured oil chitosan and three polycation-modified materials, as well as amount of differentiation in PC12 cells on these substrates. RESULTS: (1) Morphology of mouse Schwann cells and PC 12 cells on all substrates: after 3 days in culture, oil three different chitosan/polycation composite Substrates, Schwann cells were connected to each other and exhibited greater proliferation, compared to the chitosan control. In particular, on PLL-0.25 and POR-0.25 Substrates, some cells congregated and nearly reached confluence. The PC12 cells oil chitosan substrate, after 3 days in Culture, maintained a round shape; few exhibited a bipolar shape and began to form neurite extensions. However. on PLL-0.25 and POR-0.25 substrates, most PC12 cells displayed I bipolar shape with obvious neurite outgrowth, and almost grew as an adherent, spreading monolayer. (2) Proliferation of mouse Schwann cells and PC12 cells oil all substrates: on the first day, Schwann cell proliferation oil the three composite substrates was significantly greater than the cells oil chitosan control (P<0.01). After 3 and 5 days in Culture, PLL-0.25 and POR-0.25 substrates resulted in greater cell proliferation when compared to pure chitosan (P<0.01). Oil the third and fifth day in Culture PC12 cell proliferation on PLL-0.25 and POR-0.25 was significantly greater than on chitosan substrate (P<0.01). (3) Differentiation of PC12 cells on all substrates: at all time points, the average neurite length Of Cells Cultured on composite materials was significantly longer than on chitosan control (P<0.05-0.01). Cells oil PLL-0.25 exhibited the longest average neurite length at days 3 and 5. CONCLUSION: Mouse Schwann cells and PC12 cells exhibit in vitro biocompatibility with poly-L-lysine- and poly-L-ornithine-modified substrates, which indicates that these substrates could serve as suitable Substrates for peripheral nerve regeneration.
语种	英语 ; 英语
出版者	SHENYANG EDITORIAL DEPT NEURAL REGENERATION RES ; SHENYANG ; PO BOX 1234, SHENYANG, LIAONING 110004, PEOPLES R CHINA
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/26605]
专题	清华大学
推荐引用方式 GB/T 7714	Zheng, Zhenhuan,Wei, Yujun,Wang, Gan,et al. In vitro biocompatibility of three chitosan/polycation composite materials for nerve regeneration[J],2010, 2010.
APA	Zheng, Zhenhuan,Wei, Yujun,Wang, Gan,Gong, Yandao,&Zhang, Xiufang.(2010).In vitro biocompatibility of three chitosan/polycation composite materials for nerve regeneration..
MLA	Zheng, Zhenhuan,et al."In vitro biocompatibility of three chitosan/polycation composite materials for nerve regeneration".(2010).

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