Evaluation of Photochemistry Reaction Kinetics to Pattern Bioactive Proteins on Hydrogels for Biological Applications.
Evaluation of Photochemistry Reaction Kinetics to Pattern Bioactive Proteins on Hydrogels for Biological Applications.
复制标题
DOI：
10.1016/j.bioactmat.2017.05.005
复制
发表时间：
2018-03
影响因子：
18.9
通讯作者：
Dai G
中科院分区：
文献类型：
--
作者：
Dorsey TB;Grath A;Wang A;Xu C;Hong Y;Dai G
生物活性信号在细胞功能和行为中起着重要作用。在大多数生物学研究中，可溶性生化线索，如生长因子或细胞因子，直接添加到培养基中，以维持和/或操纵体外细胞活性。然而，这些方法不能准确地模拟某些体内生物信号基序，这些基序通常固定在细胞外基质上，并且还显示对组织形态至关重要的空间梯度。除了生化线索，生物物理特性，如底物硬度可以影响细胞行为，但在传统的细胞培养实践中不容易操纵。光交联水凝胶的最新发展为生物应用提供了精确控制空间生化和生物物理线索的新工具，但要做到这一点，需要对各种水凝胶光化学动力学进行全面的研究，以便在保持蛋白质生物活性的同时进行彻底的光交联反应。在本文中，我们研究了几种光化学反应，并评估了关键的光化学参数，如光引发剂和紫外线（UV）暴露时间，以了解它们对非期望的蛋白质损伤和细胞死亡的独特贡献。我们的数据表明，在光反应过程中，蛋白质功能的保留和细胞健康的最小化需要仔细选择光引发剂的类型和浓度，以及紫外线照射时间。我们还开发了一种基于巯基降冰片烯化学的稳健方法，用于独立控制水凝胶刚度和空间生物活性模式。总的来说，我们重点介绍了一类生物活性水凝胶，用于刚度控制和位点特异性固定化生物活性蛋白/肽，用于研究细胞行为，如细胞吸引、排斥和干细胞命运。降冰片烯部分提供比其他部分更快和更有效的硫烯反应。生物活性保留取决于光引发剂类型、浓度和紫外线照射时间。细胞活力取决于光引发剂类型、浓度和紫外线照射次数。通过改变丙烯酸酯水凝胶的浓度，可以很容易地实现硬度的数量级变化。巯基降硼烯允许点击化学和容易独立控制的刚度和精确的表面生物活性模式。
Bioactive signals play many important roles on cell function and behavior. In most biological studies, soluble biochemical cues such as growth factors or cytokines are added directly into the media to maintain and/or manipulate cell activities in vitro. However, these methods cannot accurately mimic certain in vivo biological signaling motifs, which are often immobilized to extracellular matrix and also display spatial gradients that are critical for tissue morphology. Besides biochemical cues, biophysical properties such as substrate stiffness can influence cell behavior but is not easy to manipulate under conventional cell culturing practices. Recent development in photocrosslinkable hydrogels provides new tools that allow precise control of spatial biochemical and biophysical cues for biological applications, but doing so requires a comprehensive study on various hydrogel photochemistry kinetics to allow thorough photocrosslink reaction while maintain protein bioactivities at the same time. In this paper, we studied several photochemistry reactions and evaluate key photochemical parameters, such as photoinitiators and ultra-violet (UV) exposure times, to understand their unique contributions to undesired protein damage and cell death. Our data illustrates the retention of protein function and minimize of cell health during photoreactions requires careful selection of photoinitiator type and concentration, and UV exposure times. We also developed a robust method based on thiol-norbornene chemistry for independent control of hydrogel stiffness and spatial bioactive patterns. Overall, we highlight a class of bioactive hydrogels to stiffness control and site specific immobilized bioactive proteins/peptides for the study of cellular behavior such as cellular attraction, repulsion and stem cell fate. Norbornene moiety offers faster and more efficient thiol-ene reactions than others. Bioactivity retention relies on photoinitiator type, concentration and UV times. Cell viability relies on photoinitiator type, concentration and UV times. Orders of magnitude changes in stiffness can be easily achieved by varying concentrations of acrylate hydrogel. Thiol-norborene allows click chemistry and easy independent control of stiffness and precise surface bioactive patterns.
登录
查看更多内容
影响因子：
38.3
作者：
通讯作者：
--
影响因子：
3.8
作者：
Tibbitt, Mark W.;Anseth, Kristi S.
通讯作者：
Anseth, Kristi S.
影响因子：
14
作者：
Ouasti, Sihem;Donno, Roberto;Tirelli, Nicola
通讯作者：
Tirelli, Nicola
影响因子：
3.3
作者：
Igwe, John C.;Mikael, Paiyz E.;Nukavarapu, Syam P.
通讯作者：
Nukavarapu, Syam P.
影响因子：
14
作者：
Nakajima, Masafumi;Ishimuro, Toshinari;Iwata, Hiroo
通讯作者：
Iwata, Hiroo