Photomediated oxime ligation as a bioorthogonal tool for spatiotemporally-controlled hydrogel formation and modification.
Photomediated oxime ligation as a bioorthogonal tool for spatiotemporally-controlled hydrogel formation and modification.
复制标题
DOI：
10.1039/c6tb03400d
复制
发表时间：
2017-06-21
期刊：
影响因子：
--
通讯作者：
DeForest CA
中科院分区：
文献类型：
--
作者：
Farahani PE ;Adelmund SM ;Shadish JA ;DeForest CA
点击化学已被证明是用于3D细胞培养的生物相容性水凝胶形成的有价值的工具，这是由于其在生理条件下的生物正交性质和高效率。虽然传统的点击反应可以很容易地用于创建关于活细胞的均匀的功能材料，它们的自发性禁止时空控制的材料性能，从而限制了它们的效用，在重演的动态异质性特征的体内微环境。基于光聚合的技术获得了这种所需水平的4D可编程性，但通常以引入易于与生物系统发生非特异性反应的传播自由基为代价。在这里，我们提出了一种生物正交水凝胶的形成和修改，不依赖于传播自由基，通过肟连接由photocaged烷氧基胺缓和进行的策略。在温和的近UV光暴露时，光笼被裂解，释放烷氧基胺并允许与醛局部缩合。多臂交联剂，与苯甲醛或photocaged烷氧基胺官能化，形成肟基水凝胶在几分钟内曝光的活细胞的存在下。这些凝胶的聚合速率和最终的机械性能可以通过改变交联剂浓度，光强度，苯胺催化剂当量，和pH值系统地调整。此外，水凝胶的几何形状和最终的机械性能分别由紫外线照射的位置和程度控制。然后将光介导的肟连接转化为水凝胶的生化改性，其中将含有光笼化烷氧基胺的全长蛋白质固定在暴露于UV光的用户定义区域中。由光介导的肟连接提供的可编程性可以再现天然细胞外基质的动态各向异性机械和生物化学方面。因此，光聚合肟基水凝胶有望通过作为改进的4D细胞培养平台来增强对细胞-基质相互作用的理解。
Click chemistry has proved a valuable tool in biocompatible hydrogel formation for 3D cell culture, owing to its bioorthogonal nature and high efficiency under physiological conditions. While traditional click reactions can be readily employed to create uniform functional materials about living cells, their spontaneity prohibits spatiotemporal control of material properties, thereby limiting their utility in recapitulating the dynamic heterogeneity characteristic of the in vivo microenvironment. Photopolymerization-based techniques gain this desired level of 4D programmability, but often at the expense of introducing propagating free radicals that are prone to non-specific reactions with biological systems. Here we present a strategy for bioorthogonal hydrogel formation and modification that does not rely on propagating free radicals, proceeding through oxime ligation moderated by a photocaged alkoxyamine. Upon mild near UV light exposure, the photocage is cleaved, liberating the alkoxyamine and permitting localized condensation with an aldehyde. Multi-arm crosslinkers, functionalized with either benzaldehydes or photocaged alkoxyamines, formed oxime-based hydrogels within minutes of light exposure in the presence of live cells. Polymerization rates and final mechanical properties of these gels could be systematically tuned by varying crosslinker concentrations, light intensity, aniline catalyst equivalents, and pH. Moreover, hydrogel geometry and final mechanical properties were controlled by the location and extent of UV exposure, respectively. Photomediated oxime ligation was then translated to the biochemical modification of hydrogels, where full-length proteins containing photocaged alkoxyamines were immobilized in user-defined regions exposed to UV light. The programmability afforded by photomediated oxime ligation can recapitulate dynamically anisotropic mechanical and biochemical aspects of the native extracellular matrix. Consequently, photopolymerized oxime-based hydrogels are expected to enable an enhanced understanding of cell-matrix interactions by serving as improved 4D cell culture platforms.
登录
查看更多内容
DOI：
10.1002/adma.201303680
发表时间：
2014-02-12
期刊：
Advanced materials (Deerfield Beach, Fla.)
影响因子：
--
作者：
McKinnon DD;Domaille DW;Cha JN;Anseth KS
通讯作者：
Anseth KS
影响因子：
5.5
作者：
PATEL, SK;MALONE, S;COLBY, RH
通讯作者：
COLBY, RH
影响因子：
46.9
作者：
Pédelacq, JD;Cabantous, S;Waldo, GS
通讯作者：
Waldo, GS
DOI：
10.1073/pnas.0737381100
发表时间：
2003-04-29
影响因子：
11.1
作者：
Lutolf, MP;Lauer-Fields, JL;Hubbell, JA
通讯作者：
Hubbell, JA
影响因子：
15
作者：
Dirksen, Anouk;Dirksen, Sjoerd;Dawson, Philip E.
通讯作者：
Dawson, Philip E.