Krymchenko Roman, Pfirrmann Maren, van der Leeuw Sjoerd, Avila-Martinez Nancy, Versteeg Elly M M, Meuwese Rob T C, Vlig Marcel, Verdoes Martijn, Boekema Bouke K H L, van Kuppevelt Toin H, Daamen Willeke F
Radboud university medical center, Research Institute for Medical Innovation, Department of Medical BioSciences, Nijmegen, the Netherlands.
Alliance of Dutch Burn Care, Burn Research Lab, Beverwijk, the Netherlands.
Int J Biol Macromol. 2025 Jun;315(Pt 1):144548. doi: 10.1016/j.ijbiomac.2025.144548. Epub 2025 May 21.
Elastin and elastin-derived compounds are promising biomaterials due to their biological activity, unique natural crosslinks, and ability to mimic native tissue properties. Solubilized elastin peptides retain the bioactivity of elastin and are more suitable for wound care applications than the insoluble form. Chemically solubilized elastins have shown advantageous effects in skin regeneration in humans. Here, five solubilized elastins were prepared via chemical (stepwise and continuously hydrolyzed with oxalic acid - OxA-st-ELN and OxA-ELN, or with potassium hydroxide - KOH-ELN), enzymatic (Enz-ELN), or combined (Combi-ELN) methods. OxA-st-ELN had the largest molecular weights (MWs) fragments, while Enz-ELN and Combi-ELN yielded the smallest. The effects of elastin preparations were evaluated on primary human cells - dermal fibroblasts and macrophages. In fibroblast assays, Enz-ELN induced elastin, collagen, and fibrillin-2 protein deposition, while other preparations exhibited levels comparable to the control. α-smooth muscle actin (SMA) expression remained low across all conditions. Continuous oxalic acid hydrolysis simplified the traditional stepwise approach while maintaining bioactivity. Macrophage studies showed chemical hydrolysates preserved the M0-like subtype, while Enz-ELN promoted a pro-inflammatory M1-like phenotype, and Combi-ELN had mixed effects. OxA-ELN and KOH-ELN appeared to be the most promising options for developing biomaterial dermal scaffolds that support tissue regeneration in vivo.
弹性蛋白及弹性蛋白衍生化合物因其生物活性、独特的天然交联结构以及模拟天然组织特性的能力，成为很有前景的生物材料。可溶性弹性蛋白肽保留了弹性蛋白的生物活性，比不溶性形式更适合用于伤口护理。化学溶解的弹性蛋白已在人类皮肤再生中显示出有益效果。在此，通过化学方法（用草酸逐步和连续水解——草酸逐步水解弹性蛋白（OxA-st-ELN）和草酸水解弹性蛋白（OxA-ELN），或用氢氧化钾——氢氧化钾水解弹性蛋白（KOH-ELN））、酶解方法（酶解弹性蛋白（Enz-ELN））或联合方法（联合水解弹性蛋白（Combi-ELN））制备了五种可溶性弹性蛋白。OxA-st-ELN具有最大分子量片段，而Enz-ELN和Combi-ELN产生的片段最小。评估了弹性蛋白制剂对原代人细胞——真皮成纤维细胞和巨噬细胞的影响。在成纤维细胞试验中，Enz-ELN诱导了弹性蛋白、胶原蛋白和原纤维蛋白-2的蛋白沉积，而其他制剂表现出与对照相当的水平。在所有条件下，α-平滑肌肌动蛋白（SMA）表达均保持较低水平。连续草酸水解简化了传统的逐步方法，同时保持了生物活性。巨噬细胞研究表明，化学水解产物保留了M0样亚型，而Enz-ELN促进了促炎性M1样表型，Combi-ELN具有混合效应。OxA-ELN和KOH-ELN似乎是开发支持体内组织再生的生物材料真皮支架最有前景的选择。
