Dual alginate crosslinking for local patterning of biophysical and biochemical properties.
Dual alginate crosslinking for local patterning of biophysical and biochemical properties.
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DOI：
10.1016/j.actbio.2020.07.047
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发表时间：
2020-07
影响因子：
9.7
通讯作者：
Aline Lueckgen;D. S. Garske;A. Ellinghaus;D. Mooney;G. Duda;A. Cipitria
中科院分区：
文献类型：
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作者：
Aline Lueckgen;D. S. Garske;A. Ellinghaus;D. Mooney;G. Duda;A. Cipitria
具有模式化生物物理和生物化学性质的水凝胶在生物材料界受到越来越多的关注。在这项工作中，我们探索了两种正交交联机制的海藻酸盐基材料：自发的Diels-Alder反应和紫外光引发的硫醇-烯反应。将这些机制结合在一种材料中并使用光掩模在空间上限制后者的位置，使得能够形成具有刚度、生物分子呈现和降解模式的双交联水凝胶，从而对细胞行为进行局部控制。在刚度图案的特征在于形态学上的共聚焦显微镜和机械单轴压缩和微压痕测量。小鼠胚胎成纤维细胞接种在刚性图案基板上附着优选，并获得一个蔓延的形态硬相比，软的地区。人骨髓间充质干细胞在软表面上表现出优先的成脂分化，在硬表面上表现出优先的成骨分化。生物分子呈递的模式揭示了小鼠前成骨细胞在条纹区域上的有利附着，其中硫醇化细胞粘附生物分子已经偶联。在胶原酶暴露后，通过显微压痕测量可视化降解模式。当将这些材料皮下植入小鼠背部时，在n=5/12的样品中可辨别出图案化组织浸润到表面上的可降解区域中。综合起来看，这些结果表明，具有生物物理和生物化学性质模式的水凝胶系统能够研究环境因素如何影响体外多种细胞行为，并且可以应用于指导体内不同愈合场景中的内源性组织生长。细胞行为在我们的基于藻酸盐的系统中，我们证明了局部基底刚度和生物分子呈递对两种不同小鼠细胞系和人原代细胞的体外细胞附着、形态、迁移和分化行为的影响。此外，在皮下植入小鼠模型后，分析了降解模式对体内组织浸润的影响。在水凝胶模板之后实现图案化的组织浸润代表了朝向引导内源性愈合反应的重要步骤，从而邀请在各种组织工程背景中的应用。
Hydrogels with patterned biophysical and biochemical properties have found increasing attention in the biomaterials community. In this work, we explore alginate-based materials with two orthogonal crosslinking mechanisms: the spontaneous Diels-Alder reaction and the ultraviolet light-initiated thiol-ene reaction. Combining these mechanisms in one material and spatially restricting the location of the latter using photomasks, enables the formation of dual-crosslinked hydrogels with patterns in stiffness, biomolecule presentation and degradation, granting local control over cell behavior. Patterns in stiffness are characterized morphologically by confocal microscopy and mechanically by uniaxial compression and microindentation measurement. Mouse embryonic fibroblasts seeded on stiffness-patterned substrates attach preferably and attain a spread morphology on stiff compared to soft regions. Human mesenchymal stem cells demonstrate preferential adipogenic differentiation on soft surfaces and osteogenic differentiation on stiff surfaces. Patterns in biomolecule presentation reveal favored attachment of mouse pre-osteoblasts on stripe regions, where thiolated cell-adhesive biomolecules have been coupled. Patterns in degradation are visualized by microindentation measurement following collagenase exposure. Patterned tissue infiltration into degradable regions on the surface is discernible in n=5/12 samples, when these materials are implanted subcutaneously into the backs of mice. Taken together, these results demonstrate that our hydrogel system with patterns in biophysical and biochemical properties enables the study of how environmental cues affect multiple cell behaviors in vitro and could be applied to guide endogenous tissue growth in diverse healing scenarios in vivo.Statement of SignificanceHydrogels with patterns in biophysical and biochemical properties have been explored in the biomaterials community in order to spatially control or guide cell behavior. In our alginate-based system, we demonstrate the effect of local substrate stiffness and biomolecule presentation on the in vitro cell attachment, morphology, migration and differentiation behavior of two different mouse cell lines and human primary cells. Additionally, the effect of degradation patterns on the in vivo tissue infiltration is analyzed following subcutaneous implantation into a mouse model. The achievement of patterned tissue infiltration following the hydrogel template represents an important step towards guiding endogenous healing responses, thus inviting application in various tissue engineering contexts.