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羧甲基壳聚糖/透明质酸聚乙烯醇共混膜的制备及其理化性质

[Preparation and physicochemical property of carboxymethyl-chitosan/hyaluronic acid poly(vinyl alcohol) blend membrane].

作者信息

Liu Wen, Li Shuning, Chang Jing, Han Baoqin, Liu Wanshun

机构信息

College of Marine Life Science, Ocean University of China, Qingdao Shandong 266003, PR China.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2009 Aug;23(8):1012-6.

PMID:19728624
Abstract

OBJECTIVE

To prepare carboxymethyl-chitosan/hyaluronic acid/poly(vinyl alcohol) (CHP) blend membrane, evaluate its physicochemical properties and intraocular biocompatibility and to investigate its feasibility to be applied to glaucoma filtering surgery.

METHODS

CHP blend membrane was prepared using solution casting method after blending carboxymethyl-chitosan, HA and poly(vinyl alcohol) in a proportion of 5 : 4 : 1 (M/M). Its water absorption rate, swelling rate, permeability, and mechanical properties were detected. Subconjunctival fibroblasts separated from subconjunctival tissue of New Zealand white rabbits were cultured, and the cells at passage 4 were cultured on cell culture plate with or without the CHP blend membrane, serving as the experimental group and the control group, respectively. Effect of the CHP blend membrane on the subconjunctival fibroblasts was tested by MTT method 24, 48, and 72 hours after culture. Six New Zealand white rabbits were randomly divided into two groups (n = 3 rabbits per group), and the CHP blend membrane and SK gel were implanted into the rabbits' subconjunctival space and anterior chamber in the experimental group and the control group, respectively. Slit lamp observation and binocular reaction record were conducted 1, 3, 5, 9, 11, 20, 30, 45, and 60 days after operation. Corneal tissue harvested from the experimental group was observed using scanning electron microscope 15 days after operation to study ophthalmic biocompatibility and biodegradability.

RESULTS

The water absorption rate and the swelling rate of the CHP blend membrane was 83.8% +/- 1.3% and 3.59 +/- 0.50, respectively. The tensile strength of the dry and the wet CHP blend membrane was (20.59 +/- 1.73) and (0.51 +/- 0.13) MPa, respectively. The breaking elongation rate of the dry and the wet CHP blend membrane was 10.69% +/- 1.16% and 53.15% +/- 2.46%, respectively. The CHP blend membrane had good permeability to NaCl and L-tyrosine. Absorbance (A) value of the experimental group 24, 48, and 72 hours after breeding was 0.207 +/- 0.083, 0.174 +/- 0.080, and 0.181 +/- 0.048, respectively, while the A value of the control group was 0.284 +/- 0.011, 0.272 +/- 0.083, and 0.307 +/- 0.056, respectively. Significant difference was evident between two groups (P < 0.05). In the experimental group, a small amount of floccus was exuded around the implanted membrane 1 day after operation; the floccus was absorbed on the third day, and there was no obvious inflammatory reaction occurring on the eleventh day. Most of the membrane degraded on the sixtieth day. Scanning electron microscope observation showed that the hexagonal morphology of the corneal endothelial cells was intact, and no degradation particles adhered to the surface. In the control group, the implantation of SK gel into anterior chamber was unsuccessful because the SK gel was quite soft and easily broken. In the experimental group, mild hyperemia emerged around the implanted membrane 1 day after the subconjunctival implantation of the membrane, and it became normal on the ninth day. No corneal edema and inflammatory reaction of anterior chamber occurred till the sixtieth day. The results in the control group and the experiment group were similar.

CONCLUSION

Due to its good physicochemical properties and biocompatibility, the CHP blend membrane has potential applications in glaucoma filtering surgery.

摘要

目的

制备羧甲基壳聚糖/透明质酸/聚乙烯醇(CHP)共混膜,评估其理化性质和眼内生物相容性,并探讨其应用于青光眼滤过手术的可行性。

方法

按5∶4∶1(M/M)的比例将羧甲基壳聚糖、透明质酸和聚乙烯醇混合后采用溶液浇铸法制备CHP共混膜。检测其吸水率、溶胀率、通透性及力学性能。分离新西兰白兔结膜下组织中的结膜下成纤维细胞进行培养,将第4代细胞分别接种于有或无CHP共混膜的细胞培养板上,分别作为实验组和对照组。培养24、48和72小时后,采用MTT法检测CHP共混膜对结膜下成纤维细胞的影响。将6只新西兰白兔随机分为两组(每组n = 3只),实验组和对照组分别将CHP共混膜和SK凝胶植入兔结膜下间隙和前房。术后1、3、5、9、11、20、30、45和60天进行裂隙灯观察并记录双眼反应。术后15天取实验组角膜组织进行扫描电子显微镜观察,研究眼内生物相容性和生物降解性。

结果

CHP共混膜的吸水率和溶胀率分别为83.8%±1.3%和3.59±0.50。干燥和湿态CHP共混膜的拉伸强度分别为(20.59±1.73)和(0.51±0.13)MPa。干燥和湿态CHP共混膜的断裂伸长率分别为10.69%±1.16%和53.15%±2.46%。CHP共混膜对氯化钠和L-酪氨酸具有良好的通透性。培养24、48和72小时后实验组的吸光度(A)值分别为0.207±0.083、0.174±0.080和0.181±0.048,而对照组的A值分别为0.284±0.011、0.272±0.083和0.307±0.056。两组间差异有统计学意义(P < 0.05)。实验组术后1天植入膜周围有少量絮状物渗出,第3天絮状物吸收,第11天无明显炎症反应。第60天大部分膜降解。扫描电子显微镜观察显示角膜内皮细胞六边形形态完整,表面无降解颗粒附着。对照组将SK凝胶植入前房未成功,因为SK凝胶很软且容易破碎。实验组结膜下植入膜后1天植入膜周围出现轻度充血,第9天恢复正常。至第60天无角膜水肿和前房炎症反应发生。对照组和实验组结果相似。

结论

CHP共混膜因其良好的理化性质和生物相容性,在青光眼滤过手术中具有潜在应用价值。

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