3 years ago

Tethering Growth Factors to Collagen Surfaces Using Copper-Free Click Chemistry: Surface Characterization and in Vitro Biological Response

Tethering Growth Factors to Collagen Surfaces Using Copper-Free Click Chemistry: Surface Characterization and in Vitro Biological Response
David Myung, Ilham Putra, Gabriella M. Fernandes-Cunha, Hyun Jong Lee, Won-Gun Koh
Surface modifications with tethered growth factors have mainly been applied to synthetic polymeric biomaterials in well-controlled, acellular settings, followed by seeding with cells. The known bio-orthogonality of copper-free click chemistry provides an opportunity to not only use it in vitro to create scaffolds or pro-migratory tracks in the presence of living cells, but also potentially apply it to living tissues directly as a coupling modality in situ. In this study, we studied the chemical coupling of growth factors to collagen using biocompatible copper-free click chemistry and its effect on the enhancement of growth factor activity in vitro. We verified the characteristics of modified epidermal growth factor (EGF) using mass spectrometry and an EGF/EGF receptor binding assay, and evaluated the chemical immobilization of EGF on collagen by copper-free click chemistry using surface X-ray photoelectron spectroscopy (XPS), surface plasmon resonance (SPR) spectroscopy, and enzyme-linked immunosorbent assay (ELISA). We found that the anchoring was noncytotoxic, biocompatible, and rapid. Moreover, the surface-immobilized EGF had significant effects on epithelial cell attachment and proliferation. Our results demonstrate the possibility of copper-free click chemistry as a tool for covalent bonding of growth factors to collagen in the presence of living cells. This approach is a novel and potentially clinically useful application of copper-free click chemistry as a way of anchoring growth factors to collagen and foster epithelial wound healing.

Publisher URL: http://dx.doi.org/10.1021/acsami.7b05262

DOI: 10.1021/acsami.7b05262

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