Zhong Lin, Banigo Alma Tamunonengiofori, Zoetebier Bram, Karperien Marcel
Department of Developmental BioEngineering, Faculty of Science and Technology, TechMed Centre, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
Gels. 2024 Aug 30;10(9):566. doi: 10.3390/gels10090566.
Hydrogels are widely used in tissue engineering due to their ability to form three-dimensional (3D) structures that support cellular functions and mimic the extracellular matrix (ECM). Despite their advantages, dextran-based hydrogels lack intrinsic biological activity, limiting their use in this field. Here, we present a strategy for developing bioactive hydrogels through sequential thiol-maleimide bio-functionalization and enzyme-catalyzed crosslinking. The hydrogel network is formed through the reaction of tyramine moieties in the presence of horseradish peroxidase (HRP) and hydrogen peroxide (HO), allowing for tunable gelation time and stiffness by adjusting HO concentrations. Maleimide groups on the hydrogel backbone enable the coupling of thiol-containing bioactive molecules, such as arginylglycylaspartic acid (RGD) peptides, to enhance biological activity. We examined the effects of hydrogel stiffness and RGD concentration on human mesenchymal stem cells (hMSCs) during differentiation and found that hMSCs encapsulated within these hydrogels exhibited over 88% cell viability on day 1 across all conditions, with a slight reduction to 60-81% by day 14. Furthermore, the hydrogels facilitated adipogenic differentiation, as evidenced by positive Oil Red O staining. These findings demonstrate that DexTA-Mal hydrogels create a biocompatible environment that is conducive to cell viability and differentiation, offering a versatile platform for future tissue engineering applications.
水凝胶因其能够形成支持细胞功能并模拟细胞外基质（ECM）的三维（3D）结构而被广泛应用于组织工程。尽管具有诸多优点，但基于葡聚糖的水凝胶缺乏内在生物活性，限制了它们在该领域的应用。在此，我们提出一种通过巯基-马来酰亚胺生物功能化和酶催化交联来开发生物活性水凝胶的策略。水凝胶网络是在辣根过氧化物酶（HRP）和过氧化氢（HO）存在的情况下，通过酪胺部分的反应形成的，通过调节HO浓度可实现可调节的凝胶化时间和硬度。水凝胶主链上的马来酰亚胺基团能够偶联含巯基的生物活性分子，如精氨酰甘氨酰天冬氨酸（RGD）肽，以增强生物活性。我们研究了水凝胶硬度和RGD浓度在人骨髓间充质干细胞（hMSCs）分化过程中的作用，发现在所有条件下，封装在这些水凝胶中的hMSCs在第1天的细胞活力超过88%，到第14天略有下降至60 - 81%。此外，水凝胶促进了脂肪生成分化，油红O染色阳性证明了这一点。这些发现表明，DexTA - Mal水凝胶创造了一个有利于细胞活力和分化的生物相容性环境，为未来的组织工程应用提供了一个通用平台。
