Lequoy Pauline, Murschel Frederic, Liberelle Benoit, Lerouge Sophie, De Crescenzo Gregory
Department of Mechanical Engineering, École de technologie supérieure (ÉTS), 1100 boul. Notre-Dame Ouest, Montréal, QC H3C 1K3, Canada; Research Centre, Centre Hospitalier de l'Université de Montréal (CRCHUM), 900 St Denis, Tour Viger, Montréal, QC H2X 0A9, Canada.
Department of Chemical Engineering, École Polytechnique de Montréal, P.O. Box 6079, succ. Centre-Ville, Montréal, QC H3C 3A7, Canada.
Acta Biomater. 2016 Jan;29:239-247. doi: 10.1016/j.actbio.2015.10.026. Epub 2015 Oct 17.
Growth factors (GFs) are potent signaling molecules that act in a coordinated manner in physiological processes such as tissue healing or angiogenesis. Co-immobilizing GFs on materials while preserving their bioactivity still represents a major challenge in the field of tissue regeneration and bioactive implants. In this study, we explore the potential of an oriented immobilization technique based on two high affinity peptides, namely the Ecoil and Kcoil, to allow for the simultaneous capture of the epidermal growth factor (EGF) and the vascular endothelial growth factor (VEGF) on a chondroitin sulfate coating. This glycosaminoglycan layer was selected as it promotes cell adhesion but reduces non-specific adsorption of plasma proteins. We demonstrate here that both Ecoil-tagged GFs can be successfully immobilized on chondroitin sulfate surfaces that had been pre-decorated with the Kcoil peptide. As shown by direct ELISA, changing the incubation concentration of the various GFs enabled to control their grafted amount. Moreover, cell survival studies with endothelial and smooth muscle cells confirmed that our oriented tethering strategy preserved GF bioactivity. Of salient interest, co-immobilizing EGF and VEGF led to better cell survival compared to each GF captured alone, suggesting a synergistic effect of these GFs. Altogether, these results demonstrate the potential of coiled-coil oriented GF tethering for the co-immobilization of macromolecules; it thus open the way to the generation of biomaterials surfaces with fine-tuned biological properties.
Growth factors are potent signaling molecules that act in a coordinated manner in physiological processes such as tissue healing or angiogenesis. Controlled coimmobilization of growth factors on biomaterials while preserving their bioactivity represents a major challenge in the field of tissue regeneration and bioactive implants. This study demonstrates the potential of an oriented immobilization technique based on two high affinity peptides to allow for the simultaneous capture of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF). Our system allowed an efficient control on growth factor immobilization by adjusting the incubation concentrations of EGF and VEGF. Of salient interest, co-immobilizing of specific ratios of EGF and VEGF demonstrated a synergistic effect on cell survival compared to each GF captured alone.
生长因子（GFs）是在诸如组织愈合或血管生成等生理过程中以协同方式起作用的强效信号分子。在材料上共固定生长因子并同时保留其生物活性仍是组织再生和生物活性植入物领域中的一项重大挑战。在本研究中，我们探索了一种基于两种高亲和力肽（即Ecoil和Kcoil）的定向固定技术，以实现将表皮生长因子（EGF）和血管内皮生长因子（VEGF）同时捕获到硫酸软骨素涂层上的潜力。选择这种糖胺聚糖层是因为它能促进细胞黏附，但能减少血浆蛋白的非特异性吸附。我们在此证明，两种带有Ecoil标签的生长因子都能成功固定在预先用Kcoil肽修饰过的硫酸软骨素表面上。如直接酶联免疫吸附测定（ELISA）所示，改变各种生长因子的孵育浓度能够控制它们的接枝量。此外，对内皮细胞和平滑肌细胞进行的细胞存活研究证实，我们的定向连接策略保留了生长因子的生物活性。特别值得注意的是，与单独捕获每种生长因子相比，共固定EGF和VEGF导致更好的细胞存活，这表明这些生长因子具有协同效应。总之，这些结果证明了卷曲螺旋定向生长因子连接用于大分子共固定的潜力；因此，它为生成具有精细调节生物学特性的生物材料表面开辟了道路。
生长因子是在诸如组织愈合或血管生成等生理过程中以协同方式起作用的强效信号分子。在保留其生物活性的同时，对生长因子在生物材料上进行可控的共固定是组织再生和生物活性植入物领域中的一项重大挑战。本研究证明了一种基于两种高亲和力肽的定向固定技术，可实现同时捕获表皮生长因子（EGF）和血管内皮生长因子（VEGF）。我们的系统通过调整EGF和VEGF的孵育浓度，能够有效地控制生长因子的固定。特别值得注意的是，与单独捕获每种生长因子相比，以特定比例共固定EGF和VEGF对细胞存活表现出协同效应。
