Clickable decellularized extracellular matrix as a new tool for building hybrid-hydrogels to model chronic fibrotic diseases in vitro.
Clickable decellularized extracellular matrix as a new tool for building hybrid-hydrogels to model chronic fibrotic diseases in vitro.
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DOI：
10.1039/d0tb00613k
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发表时间：
2020-08-21
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影响因子：
--
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纤维化疾病占全世界死亡人数的三分之一以上。尽管人们付出了巨大努力来研究纤维化背后的细胞和分子过程，但目前有效的治疗方法很少。双阶段聚合反应是一种创新工具，用于重现细胞外基质 (ECM) 模量的异质增加，这是体内纤维化疾病的标志。在这里，我们提出了一种可点击的脱细胞 ECM (dECM) 交联剂，其结合到动态响应的聚乙二醇-α-甲基丙烯酸酯 (PEGαMA) 混合水凝胶中，以在体外重建 ECM 重塑。在 PEGαMA（8 臂，10 kg/mol）和可点击的 dECM 之间添加非化学计量的硫醇烯迈克尔，形成弹性模量为 E = 3.6 ± 0.24 kPa 的水凝胶，接近健康肺组织 (1-5 kPa)。接下来，残余的 αMA 基团通过光引发均聚反应，将模量值提高到原位纤维化水平 (E = 13.4 ± 0.82 kPa)。弹性模量增加的水凝胶模仿纤维化 ECM，诱导肌成纤维细胞转基因表达显着增加。与软水凝胶相比，在硬质和动态硬化的混合水凝胶上培养时，表达胶原蛋白 1a1 和 α-平滑肌肌动蛋白的双报告基因小鼠肺部的原代成纤维细胞比例增加了约 60%。同样，与柔软区域相比，空间图案混合水凝胶的坚硬区域上的成纤维细胞表达的胶原蛋白1a1转基因水平显着增加。总的来说，这些结果表明混合水凝胶是一种新工具，可用于在体外时空诱导原代小鼠成纤维细胞的表型转变。含有可点击的脱细胞细胞外基质的水解稳定、光可调的混合水凝胶能够对成纤维细胞活化进行时空控制。
Fibrotic disorders account for over one third of mortalities worldwide. Despite great efforts to study the cellular and molecular processes underlying fibrosis, there are currently few effective therapies. Dual-stage polymerization reactions are an innovative tool for recreating heterogeneous increases in extracellular matrix (ECM) modulus, a hallmark of fibrotic diseases in vivo. Here, we present a clickable decellularized ECM (dECM) crosslinker incorporated into a dynamically responsive poly(ethylene glycol)-α-methacrylate (PEGαMA) hybrid-hydrogel to recreate ECM remodeling in vitro. An off-stoichiometry thiol-ene Michael addition between PEGαMA (8-arm, 10 kg/mol) and the clickable dECM resulted in hydrogels with an elastic modulus of E = 3.6 ± 0.24 kPa, approximating healthy lung tissue (1-5 kPa). Next, residual αMA groups were reacted via a photo-initiated homopolymerization to increase modulus values to fibrotic levels (E = 13.4 ± 0.82 kPa) in situ. Hydrogels with increased elastic moduli, mimicking fibrotic ECM, induced a significant increase in the expression of myofibroblast transgenes. The proportion of primary fibroblasts from dual-reporter mouse lungs expressing collagen 1a1 and alpha-smooth muscle actin increased by approximately 60% when cultured on stiff and dynamically stiffened hybrid-hydrogels compared to soft. Likewise, fibroblasts expressed significantly increased levels of the collagen 1a1 transgene on stiff regions of spatially patterned hybrid-hydrogels compared to the soft areas. Collectively, these results indicate that hybrid-hydrogels are a new tool that can be implemented to spatiotemporally induce a phenotypic transition in primary murine fibroblasts in vitro. Hydrolytically stable, phototunable hybrid-hydrogels containing clickable decellularized extracellular matrix enable spatiotemporal control over fibroblast activation.
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影响因子：
9.7
作者：
Chirco KR;Worthington KS;Flamme-Wiese MJ;Riker MJ;Andrade JD;Ueberheide BM;Stone EM;Tucker BA;Mullins RF
通讯作者：
Mullins RF
影响因子：
9.7
作者：
Gould, Sarah T.;Darling, Nicole J.;Anseth, Kristi S.
通讯作者：
Anseth, Kristi S.
DOI：
10.1016/j.matbio.2017.12.004
发表时间：
2018-11
期刊：
Matrix biology : journal of the International Society for Matrix Biology
影响因子：
--
作者：
Haak AJ;Tan Q;Tschumperlin DJ
通讯作者：
Tschumperlin DJ
影响因子：
16.6
作者：
Guvendiren, Murat;Burdick, Jason A.
通讯作者：
Burdick, Jason A.
DOI：
10.1165/rcmb.2015-0095oc
发表时间：
2016-04-01
影响因子：
6.4
作者：
Green，Jenna;Endale，Mehari;Perl，Anne-Karina T.
通讯作者：
Perl，Anne-Karina T.