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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2018, Vol. 38 Issue (4): 70-77    DOI: 10.13523/j.cb.20180410
技术与方法     
负载NGF的可注射壳聚糖透明质酸水凝胶材料理化性能及生物相容性研究
段思腾1,李光然1,马义勇1,邱裕佳1,李宇2,王伟1,3,4()
1 锦州医科大学附属第一医院 锦州 121000
2 辽宁中医药大学研究生院 沈阳 110016
3 锦州医科大学骨外科学研究所 锦州 121000
4 辽宁省医学组织工程重点实验室 锦州 121000
Study on Physicochemical Properties and Biocompatibility of Injectable Chitosan-hyaluronic Acid Hydrogel Loaded with NGF
Si-teng DUAN1,Guang-ran LI1,Yi-yong MA1,Yu-jia QIU1,Yu LI2,Wei WANG1,3,4()
1 The First Affiliated Hospital, Jinzhou Medical University, Jinzhou 121000, China
2 School of Graduate, Liaoning University of Traditional Chinese Medicine, Shenyang 110016, China
3 Department of Orthopedics Research Institute, Jinzhou Medical University, Jinzhou　121000, China
4 Key Laboratory of Medical Tissue Engineering, Jinzhou　121000, China
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摘要： 目的

制备负载NGF的可注射壳聚糖透明质酸复合水凝胶,探讨其理化性能以及生物相容性。

 方法

首先京尼平交联制备壳聚糖透明质酸复合水凝胶材料,采用倒置法检测凝胶时间;扫描电镜观察材料形态结构;NGF释放实验、体外溶胀以及降解实验等检测凝胶材料的物理化学性能;通过MTT实验、NGF活性检测、材料与细胞共培养检测凝胶材料生物学性能。

 结果

在37℃条件下,可注射凝胶材料凝胶时间在37min左右,凝胶材料为多孔网络状结构,凝胶材料8周最多能够降解76%,缓释21天的NGF具有生物活性,凝胶材料能促进RSC96细胞的粘附、增殖、迁移以及细胞活性物质的释放。

 结论

京尼平交联的壳聚糖透明质酸水凝胶具有良好的生物相容性,能作为NGF的载体材料,具有成为神经导管内填充材料的潜能。
关键词： 可注射水凝胶;  壳聚糖;  透明质酸;  京尼平;  神经生长因子    
Abstract: Objective:

To prepare a carrier for NGF with injectable chitosan-hyaluronic acid hydrogel,and studying its physicochemical properties and biocompatibility.

 Methods:

Preparing chitosan-hyaluronic acid hydrogel crosslinked with genipin, inverted method was used to determine the gelation time; scanning electron microscopy was used to observe the morphological structure; the physical and chemical properties were measured by NGF release, vitro swelling and degradation assay; the biological properties were measured by MTT, NGF activity assay and Co-Culture experiment.

 Result:

Injectable hydrogel which had a structure porous network congealed at 37℃ in about 37 minutes, and its mostly degradation degree in 8 weeks was 76%, sustained release for 21 days with biological activity. Moreover, the chitosan-hyaluronic acid hydrogel could promote RSC96 adhesion, proliferation, migration and release of cell active substances.

 Conclusion:

Genipin-crosslinked chitosan-hyaluronic acid hydrogel have good biocompatibility, and can be used as a carrier of NGF. It could be used as a potential filling material for nerve conduits.
Key words: Injectable hydrogel    Chitosan    Hyaluronic acid    Genipin    Nerve growth factor
收稿日期: 2017-12-14 出版日期: 2018-05-08
ZTFLH:  Q33  
基金资助: 辽宁省自然科学基金面上项目(201602322);辽宁省教育厅科学技术研究地方服务项目(JYTFUDF201757)
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段思腾
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李宇
王伟

引用本文:

段思腾,李光然,马义勇,邱裕佳,李宇,王伟. 负载NGF的可注射壳聚糖透明质酸水凝胶材料理化性能及生物相容性研究[J]. 中国生物工程杂志, 2018, 38(4): 70-77.

Si-teng DUAN,Guang-ran LI,Yi-yong MA,Yu-jia QIU,Yu LI,Wei WANG. Study on Physicochemical Properties and Biocompatibility of Injectable Chitosan-hyaluronic Acid Hydrogel Loaded with NGF. China Biotechnology, 2018, 38(4): 70-77.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20180410    或    https://manu60.magtech.com.cn/biotech/CN/Y2018/V38/I4/70

组别 GN含量(mg) 凝胶时间(min) 孔径(μm) 孔隙率(%)
A 1 38.25±3.10 42.90±9.51 70.35±2.28
B 2 38.25±1.26 40.68±11.87 66.97±3.23
C 3 36.00±2.83 36.33±16.65 62.13±3.43
D 4 37.00±1.83 34.27±15.31 57.37±3.61
表1  不同组别水凝胶的凝胶时间、孔径、孔隙率
图1  不同凝胶材料的红外光谱图分析
图2  冻干CS-HA凝胶扫描电镜观察
图3  CS-HA溶胀比率与降解率时间动态图
图4  NGF累计释放曲线
图5  CS-HA释放NGF活性检测
图6  各组材料1、3、5天OD值
图7  RSC96与CS-HA材料共培养电镜照片
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