Photoinitiator-free synthesis of endothelial cell-adhesive and enzymatically degradable hydrogels.
Photoinitiator-free synthesis of endothelial cell-adhesive and enzymatically degradable hydrogels.
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DOI：
10.1016/j.actbio.2014.11.012
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发表时间：
2015-02
影响因子：
9.7
通讯作者：
Kottke-Marchant, Kandice
中科院分区：
文献类型：
--
作者：
Jones, Derek R.;Marchant, Roger E.;von Recum, Horst;Sen Gupta, Anirban;Kottke-Marchant, Kandice
我们报道了一种无光引发剂的合成方法，将生物活性融入聚乙二醇（PEG）水凝胶中，以控制聚合物网络的物理性质、酶生物降解性和细胞特异性粘附性，同时消除了对紫外线介导的光聚合的需要。为了实现这一目标，水凝胶网络使用四臂PEG丙烯酸酯（10 kDa）的Michael加成聚合，使用胶原酶敏感肽（CSP）作为交联剂，并在末端（RGD）或连接在交联肽序列（CSP-RGD）上引入内皮细胞粘附肽。采用核磁共振和Ellman法测定了Michael加成反应的效率。通过量化和比较各种水凝胶配方的膨胀比和杨氏模量，证明了细胞粘附性和物理性质的成功解耦。通过在胶原酶溶液（0.0 - 1.0µg/mL）中培养功能化水凝胶来建立降解谱，表明功能化水凝胶的降解速率取决于胶原酶的浓度。此外，降解速率与CSP-RGD浓度无关。比较了不同RGD浓度下细胞在功能化水凝胶上的附着和增殖，证明细胞的附着和增殖与CSP-RGD组合肽的相对含量直接相关。与uv聚合相比，使用Michael添加技术可以提高细胞活力，并通过LIVE/DEAD荧光法进行评估。这种无光引发剂的方法在制造基于水凝胶的组织工程支架方面显示出前景，这种支架允许分离的细胞粘附性和物理性质，并提供更高的细胞活力。
We report on a photoinitiator-free synthetic method of incorporating bioactivity into poly(ethylene glycol) (PEG) hydrogels in order to control physical properties, enzymatic biodegradability and cell-specific adhesiveness of the polymer network, while eliminating the need for UV-mediated photopolymerization. To accomplish this, hydrogel networks were polymerized using Michael addition with four-arm PEG acrylate (10 kDa), using a collagenase sensitive peptide (CSP) as a crosslinker, and introducing an endothelial cell adhesive peptide either terminally (RGD) or attached to the crosslinking peptide sequence (CSP-RGD). The efficiency of the Michael addition reactions were determined by NMR and Ellman’s assay. Successful decoupling of cell adhesivity and physical properties was demonstrated by quantifying and comparing the swelling ratios and Young’s Moduli of various hydrogel formulations. Degradation profiles were established by incubating functionalized hydrogels in collagenase solutions (0.0 – 1.0 µg/mL), demonstrating that functionalized hydrogels degraded at a rate dependent upon collagenase concentration. Moreover, it was shown that the degradation rate was independent of CSP-RGD concentration. Cell attachment and proliferation on functionalized hydrogels were compared for various RGD concentrations, providing evidence that cell attachment and proliferation were directly related to relative amounts of the CSP-RGD combination peptide. An increase in cell viability was achieved using Michael addition techniques when compared to UV-polymerization, and was assessed by a LIVE/DEAD fluorescence assay. This photoinitiator-free method shows promise in creating hydrogel-based tissue engineering scaffolds allow for decoupled cell adhesivity and physical properties and that render greater cell viability.
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DOI：
10.1083/jcb.127.2.537
发表时间：
1994-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Re F;Zanetti A;Sironi M;Polentarutti N;Lanfrancone L;Dejana E;Colotta F
通讯作者：
Colotta F
影响因子：
14
作者：
Takayama, Toshio;Taguchi, Tetsushi;Nagawa, Hirokazu
通讯作者：
Nagawa, Hirokazu
水凝胶组织工程支架的设计特性。
影响因子：
3.1
作者：
Zhu J;Marchant RE
通讯作者：
Marchant RE
影响因子：
4.9
作者：
Beamish, Jeffrey A.;Zhu, Junmin;Marchant, Roger E.
通讯作者：
Marchant, Roger E.
DOI：
10.1163/156856203763572716
发表时间：
2003-01-01
影响因子：
3.6
作者：
Watkins, AW;Anseth, KS
通讯作者：
Anseth, KS