Nano Drug Delivery Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
J Mater Sci Mater Med. 2019 Sep 11;30(9):107. doi: 10.1007/s10856-019-6309-8.
In the present study, collagen hydrogel containing naringin was fabricated, characterized and used as the scaffold for peripheral nerve damage treatment. The collagen was dissolved in acetic acid, naringin added to the collagen solution, and cross-linked with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide powder (EDC; 0.10 mM) to form the hydrogel. The microstructure, swelling behavior, biodegradation, and cyto/hemocompatibility of the fabricated hydrogels were assessed. Finally, the healing efficacy of the prepared collagen hydrogel loaded with naringin on the sciatic nerve crush injury was assessed in the animal model. The characterization results showed that the fabricated hydrogels have a porous structure containing interconnected pores with the average pore size of 90 µm. The degradation results demonstrated that about 70% of the primary weight of the naringin loaded hydrogel had been lost after 4 weeks of storage in PBS. The in vitro study showed that the proliferation of Schwann cells on the collagen/naringin hydrogel was higher than the control group (tissue culture plate) at both 48 and 72 h after cell seeding and even significantly higher than pure collagen 72 h after cell seeding (*p < 0.005, **p < 0.001). The animal study implied that the sciatic functional index reached to -22.13 ± 3.00 at the end of 60th days post-implantation which was statistically significant (p < 0.05) compared with the negative control (injury without the treatment) (-82.60 ± 1.06), and the pure collagen hydrogel (-59.80 ± 3.20) groups. The hot plate latency test, the compound muscle action potential, and wet weight-loss of the gastrocnemius muscle evaluation confirmed the positive effect of the prepared hydrogels on the healing process of the induced nerve injury. In the final, the histopathologic examinations depicted that the collagen/naringin hydrogel group reduced all the histological changes induced from the nerve injury and showed more resemblance to the normal sciatic nerve, with well-arranged fibers and intact myelin sheath. The overall results implied that the prepared collagen/naringin hydrogel can be utilized as a sophisticated alternative to healing peripheral nerve damages.
在本研究中,制备了含有柚皮苷的胶原水凝胶,并对其进行了表征,用作周围神经损伤治疗的支架。将胶原溶解在乙酸中,将柚皮苷加入到胶原溶液中,然后用 1-乙基-3-(3-二甲基氨基丙基)-碳化二亚胺粉末(EDC;0.10mM)交联形成水凝胶。评估了所制备的水凝胶的微观结构、溶胀行为、生物降解性和细胞/血液相容性。最后,在动物模型中评估了柚皮苷负载胶原水凝胶对坐骨神经挤压损伤的治疗效果。表征结果表明,所制备的水凝胶具有多孔结构,含有相互连接的孔,平均孔径为 90μm。降解结果表明,在 PBS 中储存 4 周后,柚皮苷负载水凝胶的初始重量约有 70%损失。体外研究表明,在细胞接种后 48 和 72 小时,雪旺细胞在胶原/柚皮苷水凝胶上的增殖高于对照组(细胞培养板),甚至在细胞接种后 72 小时明显高于纯胶原(*p<0.005,**p<0.001)。动物研究表明,在植入后第 60 天,坐骨功能指数达到-22.13±3.00,与阴性对照组(无治疗的损伤)(-82.60±1.06)和纯胶原水凝胶(-59.80±3.20)组相比具有统计学意义(p<0.05)。热板潜伏期试验、复合肌肉动作电位和腓肠肌湿重损失评估证实了所制备的水凝胶对诱导的神经损伤愈合过程的积极作用。最后,组织病理学检查表明,胶原/柚皮苷水凝胶组减轻了神经损伤引起的所有组织学变化,与正常坐骨神经更为相似,纤维排列整齐,髓鞘完整。总体结果表明,所制备的胶原/柚皮苷水凝胶可作为治疗周围神经损伤的一种复杂替代物。