Abdel-Mageed Heidi Mohamed, Abd El Aziz Amira Emad, Abdel Raouf Batoul Mohamed, Mohamed Saleh Ahmed, Nada Dina
Molecular Biology Department, National Research Centre, El Behoth St, Dokki, Cairo, Egypt.
Centre of Excellence, Arab Academy for Science and Technology and Maritime Transport, Alexandria, Egypt.
3 Biotech. 2022 Mar;12(3):73. doi: 10.1007/s13205-022-03131-4. Epub 2022 Feb 20.
Hydrogel-based matrix prepared using biopolymers is a new frontier of emerging platforms for enzyme immobilization for biomedical applications. Catalase (CAT) delivery can be effective in inhibiting reactive oxygen species (ROS)-mediated prolongation of the wound healing process. In this study, to improve CAT stability for effective application, gelatin(Gel)-alginate (Alg) biocompatible hydrogel (Gel-Alg), as immobilization support, was prepared using calcium chloride as an ionic cross-linker. High entrapment efficiency of 92% was obtained with 2% Gel and 1.5% Alg. Hydrogel immobilized CAT (CAT-Gel-Alg) showed a wide range of pH from 4 to 9 and temperature stability between 20 to 60 °C, compared to free CAT. CAT-Gel-Alg kinetic parameters revealed an increased (24.15 mM) and a decreased (1.39 µmol HO/mg protein min) × 10. CAT-Gel-Alg retained 52% of its original activity after 20 consecutive catalytic runs and displayed improved thermal stability with a higher value (half-life of 100.43 vs. 46 min). In addition, 85% of the initial activity was maintained after 8 weeks' storage at 4 °C. At 24 h after thermal injury, a statistically significant difference in lesion sizes between the treated group and the control group was reported. Finally, our findings suggest that the superior CAT-Gel-Alg stability and reusability are resonant features for efficient biomedical applications, and ROS scavenging by CAT in the post-burn phase offers protection for local treatment of burned tissues with encouraging wound healing kinetics.
使用生物聚合物制备的水凝胶基基质是用于生物医学应用的酶固定化新兴平台的一个新前沿领域。过氧化氢酶(CAT)递送可有效抑制活性氧(ROS)介导的伤口愈合过程延长。在本研究中,为提高CAT稳定性以实现有效应用,使用氯化钙作为离子交联剂制备了明胶(Gel)-海藻酸盐(Alg)生物相容性水凝胶(Gel-Alg)作为固定化载体。2%的Gel和1.5%的Alg获得了92%的高包封率。与游离CAT相比,水凝胶固定化CAT(CAT-Gel-Alg)在4至9的广泛pH范围内以及20至60°C的温度下具有稳定性。CAT-Gel-Alg动力学参数显示 增加(24.15 mM)且 降低(1.39 µmol HO/mg蛋白质分钟)×10。连续20次催化运行后,CAT-Gel-Alg保留了其原始活性的52%,并显示出更高 的值(半衰期为100.43分钟对46分钟),热稳定性得到改善。此外,在4°C储存8周后,初始活性的85%得以保持。热损伤后24小时,治疗组和对照组之间的损伤大小存在统计学显著差异。最后,我们的研究结果表明,卓越的CAT-Gel-Alg稳定性和可重复使用性是高效生物医学应用的显著特征,并且烧伤后阶段CAT清除ROS为烧伤组织的局部治疗提供了保护,伤口愈合动力学令人鼓舞。