Marjan Tuba, Owen Alyson R, Qazi Taimoor H
Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana 47906, United States.
Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, Indiana 47906, United States.
ACS Macro Lett. 2025 Aug 13:1241-1247. doi: 10.1021/acsmacrolett.5c00339.
Biomimetic culture platforms aid in understanding cell behavior and are useful for studying mechanisms of disease progression and tissue regeneration. Synthetic hydrogels are widely used for this purpose, but while they offer advantages such as tunability and mechanical stability, they lack the range of biochemical signals present in the native microenvironment. On the other hand, decellularized extracellular matrices (dECMs) retain native biochemical signals but their adoption as stable culture platforms is limited due to batch variability, poor mechanical stability, and limited tunability. Here we report the development of hybrid hydrogels comprising dECM and a photocurable norbornene-modified hyaluronic acid (NorHA) polymer. To overcome structural heterogeneity of dECM that inhibits its solubility, uniform gelation, and spatial uniformity during cell culture, we physically process dECM by grinding, shearing, or both, prior to incorporation within NorHA. Both processing methods reduce microscale dECM aggregation and improve physical gelation at 37 °C. The addition of dECM up to 10 mg/mL within NorHA hydrogels neither affects rapid UV cross-linking nor compromises mechanical properties, as evaluated using oscillatory shear rheology and uniaxial compression testing. Both processes significantly improve the uniform distribution of dECM within 3D hybrid hydrogels, as evaluated by staining hydrogel cryosections. Fibroblasts show significantly higher spreading area and proliferation on hybrid hydrogels compared with control NorHA hydrogels. Taken together, photocurable hybrid hydrogels having uniformly distributed dECM combine the biochemical complexity of native dECM with the tunability of a synthetic polymer and represent an advance in the engineering of biomimetic platforms to investigate cell-matrix interactions.
仿生培养平台有助于理解细胞行为，对于研究疾病进展机制和组织再生非常有用。合成水凝胶广泛用于此目的，虽然它们具有诸如可调性和机械稳定性等优点，但它们缺乏天然微环境中存在的一系列生化信号。另一方面，脱细胞细胞外基质（dECM）保留了天然生化信号，但由于批次变异性、机械稳定性差和可调性有限，其作为稳定培养平台的应用受到限制。在此，我们报告了一种由dECM和光固化降冰片烯修饰的透明质酸（NorHA）聚合物组成的混合水凝胶的开发。为了克服抑制其溶解性、均匀凝胶化和细胞培养过程中空间均匀性的dECM结构异质性，我们在将dECM掺入NorHA之前，通过研磨、剪切或两者结合的方式对其进行物理处理。两种处理方法都减少了微观尺度上的dECM聚集，并改善了37℃下的物理凝胶化。在NorHA水凝胶中加入高达10mg/mL的dECM，既不影响快速紫外线交联，也不损害机械性能，这通过振荡剪切流变学和单轴压缩试验进行评估。通过对水凝胶冷冻切片进行染色评估，两种处理方法都显著改善了dECM在三维混合水凝胶中的均匀分布。与对照NorHA水凝胶相比，成纤维细胞在混合水凝胶上显示出明显更大的铺展面积和增殖。综上所述，具有均匀分布dECM的光固化混合水凝胶将天然dECM的生化复杂性与合成聚合物的可调性结合在一起，代表了在研究细胞-基质相互作用的仿生平台工程方面的一项进展。
