贻贝启发功能改性的明胶基海绵止血材料的制备与性能

doi:10.19894/j.issn.1000-0518.210050

摘要/Abstract

摘要：

由战争、手术等而造成的大出血通常会导致更大的伤痛或更高的死亡率，因此，非常需要及时有效的止血以减少创伤导致死亡。而目前的止血材料都存在止血速度慢、止血效果差等问题。为提高材料的止血效率，本文受贻贝启发使用多巴胺和赖氨酸接枝改性的明胶（GDL）和氧化葡聚糖（ODE）为原料，通过冷冻干燥法制备多孔海绵状止血材料（GDL／ODE）。结果显示，由于多巴胺的粘附作用与赖氨酸的粘附增强作用，GDL／ODE具有良好的组织粘附性以及可使材料在出血部位形成物理屏障的能力，在60 s后能有效止血。GDL／ODE止血海绵同时具有优异的细胞相容性和血液相容性，高吸水率和高孔隙率，这些结果表明GDL／ODE有望作为一种新型的止血材料，在生物医用材料领域有潜在的应用价值。

关键词: 功能改性, 明胶基海绵, 多孔结构, 止血材料

Abstract:

Uncontrollable hemorrhage induced by war or surgery is closely related to pain and high risk of death. Therefore, a timely and effective hemostatic material is highly required for life-threatening traumatic bleeding. However, current hemostatic materials still have various defects and side effects such as slow hemostasis and secondary infection. Herein, gelatin-dopamine-lysine (GDL) and oxidized dextran (ODE) were used to prepare sponge-like hemostasis materials through lyophilization. The results show that GDL/ODE is able to integrate with biological tissue and form physical barriers, which stops bleeding after 60 s of application. GDL/ODE composites show good cyto/hemo-compatibility and highly porous microscopic morphology and water absorption ratio. These results suggest that the developed GDL/ODE sponge is a promising multifunctional hemostasis material for biomedical fields.

Key words: Functional modification, Gelatin derivative sponge, Porous structure, Hemostasis materials

中图分类号:

O636

章朱迎, 杜德焰, 周家华, 施冬健, 陈明清. 贻贝启发功能改性的明胶基海绵止血材料的制备与性能[J]. 应用化学, 2022, 39(02): 247-257.

Zhu-Ying ZHANG, De-Yan DU, Jia-Hua ZHOU, Dong-Jian SHI, Ming-Qing CHEN. Preparation and Properties of Mussel-Inspired Modified Gelatin-Based Hemostatic Sponge[J]. Chinese Journal of Applied Chemistry, 2022, 39(02): 247-257.

图/表 7

图1 止血材料的制备示意图

Fig.1 Schematic illustration of fabrication of sponges

图2 （A）GDL的合成示意图；（B）GDL-H（a）、GDL-L（b）和Gel（c）紫外吸收光谱；（C）GDL-H（a）、GDL-L（b）和Gel（c）的核磁共振谱图；（D）ODE的合成示意图；（E）Dextran（a）和ODE（b）的核磁共振谱图

Fig.2 （A）Schematic illustration of synthesis of GDL.（B）UV absorption and（C）1H NMR spectra of GDL-H（a），GDL-L（b）and Gel（c）.（D）Schematic illustration of the synthesis of ODE and（E）1H NMR spectra of dextran（a）and ODE（b）

表1 不同投料比制备的GDL中多巴胺和赖氨酸的接枝率

Table 1 Dopamine and lysine contents of GDL with different feeding ratios

样品	明胶质量	多巴胺的物质的量	赖氨酸的物质的量	多巴胺含量	赖氨酸含量
Sample	m（Gelatin）／g	n（DA）／mmol	n（Lys）／mmol	DA content／（μmol·g^－1） ^a	Lys content／（μmol·g^－1） ^b
GDL-L	5.00	19.50	19.50	30.18	49.80
GDL-H	5.00	39.00	39.00	36.56	57.61

图3 （A）止血材料内部形貌的SEM图；（B）止血材料的孔隙率；（C）止血材料的吸水率；（D）液滴完全扩散所需时间；（E）GDL-H4／ODE1的接触角测试过程

Fig.3 （A）SEM images of internal morphologies of hemostasis sponges.（B）Porosities of hemostasis sponges.（C）Water absorptions of hemostasis sponges.（D）Time consumed for water droplets spreading out on the materials.（E）Snapshots of the droplets during the contact angle test of GDL-H4／ODE1（compared to Gel／ODE with the same ratio，*P<0.1，**P<0.01，***P<0.001；compared to pure GDL or Gel，+P<0.05，++P<0.01，+++P<0.001）

图4 （A）止血材料与猪皮粘附实验照片；（B）止血材料的粘合强度；（C）止血材料的压缩模量

Fig.4 （A）Photograph of adhesive bridging between skin substrate and sponge.（B）Adhesion strength of hemostasis sponges.（C）Compression modulus of hemostasis sponges.（compared to Gel／ODE with the same ratio，*P<0.1；compared to pure GDL or Gel，+P<0.05，++P<0.01）

图5 止血材料的细胞相容性测试：（A）细胞在材料浸提液中培养1、2和3 d后的细胞活性；（B）细胞在材料浸提液中培养1 d后的荧光图片

Fig.5 Cell compatibility of hemostasis sponges.（A）Cell viability of hemostatic sponges after cultured of 1，2 and 3 d.（B）Fluorescence images for L929 cells after cultured of 1 d.（n＝5，mean±SD，compared to Control，*P<0.05，**P<0.01，***P<0.001）

图6 材料的止血性能：（A）止血时间测试过程中血液的凝固程度；（B）止血时间；（C）凝血指数；（D）红细胞粘附率；（E）溶血率；（F）使用GDL1／ODE1材料对小鼠肝脏进行止血示意图；（G）止血海绵上血细胞粘附的SEM图像

Fig.6 Hemostatic performance of materials.（A）The state of blood after coagulation in the blood clotting time test.（B）Blood clotting time，（C）blood clotting index，and（D）adhesion of erythrocyte.（E）hemolysis rate.（F）Photographs of the livers of mice during the liver injury model of hemostasis.（G）SEM images of cells adhesion on the sponges.（*P<0.05，**P<0.01，***P<0.001）

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