Development and analysis of a novel collagen-sodium humate composite hydrogel
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摘要: 水凝胶具有弹性高、含水量高，冷效应、保湿性强、形状多变等优点，是医用敷料的主要材料之一。将具有优良生物相容性、促细胞增殖功能的胶原(COL)与具有止血、消炎等作用的腐植酸钠(NaHA)按不同比例(COL∶NaHA)共混并采用自组装方式制备了一种新型胶原-腐植酸钠复合水凝胶，并考察两者间的相互作用及复合水凝胶的结构与性能，以期应用于医用敷料行业。NaHA不改变胶原的三股螺旋结构且两者之间存在氢键与静电作用。当COL∶NaHA ≥ 4∶6时，两者间的静电结合被NaCl所屏蔽，因此体系相容性较好；然而继续增加NaHA会引起聚沉现象。当COL∶NaHA=4∶6时，两者结合率最高，达到93.2%且相容性较好，复合水凝胶的纤维具有明显的D-周期且各方面性能最佳。NaHA的释放较缓慢，24 h后仍有约80%保留在水凝胶中；热稳定性较纯胶原提升了34.9℃；储能模量和损耗模量分别为31.89 Pa和3.99 Pa。此外，随着NaHA的加入，冻干复合水凝胶的孔径缩小、孔隙分布更加均匀；复合膜的亲水性明显提升。Abstract: Hydrogel was one of main materials for medical dressings due to the excellent performance such as high elasticity, high water content, cold effect, strong moisture retentiveness and variable shape. Collagen (COL) had good biocompatibility and can promote cell proliferation. And Sodium humate (NaHA) possessed hemostatic, anti-inflammatory and other biological functions. Therefore, a novel collagen-sodium humate composite hydrogel was prepared by mixing collagen and NaHA at various COL∶NaHA ratios and collagen self-assembly, which was expected to be applied as medical dressing. Then the interaction between collagen and NaHA and the microstructures and properties of composite hydrogels were investigated. The collagen triple helix is not affected, although hydrogen bonds and electrostatic interaction occur between collagen and NaHA. At COL∶NaHA≤4∶6, the hydrogels possess good compatibility due to the shielding effect of NaCl on electrostatic binding; however, the coagulation takes place with the further increase in NaHA content. At COL∶NaHA=4∶6, the percentage of NaHA incorporated into collagen fibrils is highest and 93.2%, and the compatibility between collagen and NaHA is good; therefore, the properties of composite hydrogel consisting of mature fibrils with D-periodicity are optimum. Moreover, about 80% of NaHA still remain in hydrogel, indicating its release is slow. The thermal stability improves by 34.9°C, and storge modulus and loss modulus are 31.89 Pa and 3.99 Pa, respectively. Furthermore, the pore sizes of the lyophilized composite hydrogels decrease and the pores become dense and well-distributed. The composite films have significant rises in hydrophilicity.
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Key words:
- collagen /
- sodium humate /
- composite hydrogel /
- microstructure /
- percentage of binding /
- control release /
- thermal stability
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图 1 NaHA浓度与吸光度关系曲线
R2—Coefficient of determination
Figure 1. Relationship curve of NaHA concentration and absorbance
图 3 COL-NaHA复合水凝胶的电泳图
Figure 3. Electrophoretic pattern of COL-NaHA composite hydrogels
图 4 COL-NaHA复合物的FITR图谱 (a) 和放大图谱 (b)
a—COL; b—COL-NaHA-82; c—COL-NaHA-64; d—COL-NaHA-55; e—COL-NaHA-46; f—COL-NaHA-28; g—NaHA
Figure 4. FITR spectra (a) and enlarge spectra (b) of COL-NaHA composites
图 5 COL-NaHA复合水凝胶的AFM图像(((a), (b)) COL; ((c), (d)) COL-NaHA-64; ((e), (f)) COL-NaHA-55; ((g), (h)) COL-NaHA-46)
Figure 5. AFM images of COL-NaHA composite hydrogels (((a), (b)) COL; ((c), (d)) COL-NaHA-64; ((e), (f)) COL-NaHA-55; ((g), (h)) COL-NaHA-46)
图 6 离心后纤维中NaHA与胶原的结合率 (a) 及在PBS中浸泡若干天后纤维中NaHA的剩余结合率 (b)
Figure 6. Percentage of NaHA incorporated into collagen fibrils after centrifugation (a) and the retained percentage of NaHA after soaking in PBS solution for several days (b)
图 7 复合水凝胶 (a) 和复合膜 (b) 中NaHA的释放率
Figure 7. Release ratios of NaHA in hydrogels (a) and films (b)
图 8 COL-NaHA复合水凝胶的储能模量G' (a) 和损耗模量G" (b)
Figure 8. Storage modulus G' (a) and loss modulus G" (b) of COL-NaHA composite hydrogels
图 10 冻干COL-NaHA复合水凝胶的DSC曲线
Figure 10. DSC thermograms of lyophilized COL-NaHA composite hydrogels
图 11 冻干COL-NaHA复合水凝胶的SEM图像：(a) COL；(b) COL-NaHA-82；(c) COL-NaHA-64；(d) COL-NaHA-55；(e) COL-NaHA-46；(f) COL-NaHA-28
Figure 11. SEM images of lyophilized COL-NaHA composite hydrogels: (a) COL; (b) COL-NaHA-82; (c) COL-NaHA-64; (d) COL-NaHA-55; (e) COL-NaHA-46; (f) COL-NaHA-28
表 1 胶原-腐植酸钠(COL-NaHA)复合水凝胶的制备配方
Table 1. Formulations of collagen-sodium humate (COL-NaHA) composite hydrogels
Sample cCOL/
(mg·mL−1)cNaHA/
(mg·mL−1)COL∶NaHA COL 5.00 0.00 10∶0 COL-NaHA-82 5.00 1.25 8∶2 COL-NaHA-64 5.00 3.33 6∶4 COL-NaHA-55 5.00 5.00 5∶5 COL-NaHA-46 5.00 7.50 4∶6 COL-NaHA-28 5.00 20.00 2∶8 Note: cCOL and cNaHA—Concentrations of collagen and NaHA in COL-NaHA hydrogels, respectively. 
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