Ifergan-Azriel Liaura, Bar-Am Orit, Saar Galit, Cohen Talia, Loebel Claudia, Burdick Jason A, Seliktar Dror
The Faculty of Biomedical Engineering, Technion-Israel Institute of Technology, Haifa 3200003, Israel.
The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, 1 Efron St., Haifa 3109601, Israel.
ACS Biomater Sci Eng. 2025 Sep 8;11(9):5586-5599. doi: 10.1021/acsbiomaterials.5c00980. Epub 2025 Aug 4.
An interpenetrating polymer network (IPN) hydrogel was developed for the three-dimensional (3D) culture of multipotent mesenchymal stromal cells (MSCs) with the aim of independently controlling cell spreading and material modulus. Based on our previous studies, we formulated a semisynthetic material composed of two networks: a covalent network of poly(ethylene glycol) (PEG)-fibrinogen (PF) and a second guest-host (GH) network of hyaluronic acid (HA) coupled to β-cyclodextrin (CD) and adamantane (Ad). The PF network provided cell attachment, precise control over modulus through the incorporation of additional PEG-diacrylate (PEG-DA) cross-linking, and proteolytic degradability. The GH-HA network contributed to the hydrogel's dynamic properties through enhanced viscoelasticity. This dynamic versatility enabled MSCs to better spread and grow in the IPN, even within highly cross-linked formulations. We also observed that the IPN facilitated significantly faster cell spreading kinetics, independent of the material modulus, when compared to single-network PF hydrogels. Hydrogel biodegradation was also characterized after subcutaneous implantation for up to 8 weeks by using MRI analysis. Increasing the PEG-DA cross-linking of the IPN significantly accelerated the in vivo bioresorption, whereas the biodegradation in single-network PF hydrogels was significantly delayed by the additional PEG-DA. We conclude that the covalent cross-links maintain the bulk structural integrity of the hydrogel, whereas the reversible GH interactions provide more localized adaptability for cell-mediated proteolysis and matrix remodeling, possibly through increased network heterogeneity. This design effectively mimics the ECM by providing a more supportive environment for encapsulated cells that allows them to adhere, spread, and proliferate, which may be useful in various MSC-based tissue engineering and regenerative medicine applications.
为了独立控制细胞铺展和材料模量，开发了一种互穿聚合物网络（IPN）水凝胶用于多能间充质基质细胞（MSC）的三维（3D）培养。基于我们之前的研究，我们制备了一种由两个网络组成的半合成材料：聚乙二醇（PEG）-纤维蛋白原（PF）的共价网络和与β-环糊精（CD）及金刚烷（Ad）偶联的透明质酸（HA）的第二个客体-主体（GH）网络。PF网络提供细胞附着，通过加入额外的聚乙二醇二丙烯酸酯（PEG-DA）交联精确控制模量，并具有蛋白水解降解性。GH-HA网络通过增强粘弹性有助于水凝胶的动态特性。这种动态多功能性使MSC能够在IPN中更好地铺展和生长，即使在高度交联的配方中也是如此。我们还观察到，与单网络PF水凝胶相比，IPN促进细胞铺展动力学显著加快，且与材料模量无关。通过磁共振成像（MRI）分析对皮下植入长达8周后的水凝胶生物降解情况进行了表征。增加IPN中PEG-DA的交联显著加速了体内生物吸收，而单网络PF水凝胶中的生物降解则因额外的PEG-DA而显著延迟。我们得出结论，共价交联维持了水凝胶的整体结构完整性，而可逆的GH相互作用为细胞介导的蛋白水解和基质重塑提供了更多局部适应性，可能是通过增加网络异质性实现的。这种设计通过为封装细胞提供更有利的环境来有效模拟细胞外基质（ECM），使细胞能够附着、铺展和增殖，这在各种基于MSC的组织工程和再生医学应用中可能是有用的。
