Enhanced Biological Response of AVS-Functionalized Ti-6Al-4V Alloy through Covalent Immobilization of Collagen.
Parsa RezvanianRafael DazaPatricia A LópezMilagros RamosDaniel González-NietoManuel ElicesGustavo V GuineaJosé Pérez-RigueiroPublished in: Scientific reports (2018)
This study presents the development of an efficient procedure for covalently immobilizing collagen molecules on AVS-functionalized Ti-6Al-4V samples, and the assessment of the survival and proliferation of cells cultured on these substrates. Activated Vapor Silanization (AVS) is a versatile functionalization technique that allows obtaining a high density of active amine groups on the surface. A procedure is presented to covalently bind collagen to the functional layer using EDC/NHS as cross-linker. The covalently bound collagen proteins are characterized by fluorescence microscopy and atomic force microscopy and their stability is tested. The effect of the cross-linker concentration on the process is assessed. The concentration of the cross-linker is optimized and a reliable cleaning protocol is developed for the removal of the excess of carbodiimide from the samples. The results demonstrate that the covalent immobilization of collagen type I on Ti-6Al-4V substrates, using the optimized protocol, increases the number of viable cells present on the material. Consequently, AVS in combination with the carbodiimide chemistry appears as a robust method for the immobilization of proteins and, for the first time, it is shown that it can be used to enhance the biological response to the material.
Keyphrases
- induced apoptosis
- atomic force microscopy
- high density
- wound healing
- single molecule
- tissue engineering
- high speed
- cell cycle arrest
- randomized controlled trial
- signaling pathway
- quantum dots
- minimally invasive
- patient safety
- high resolution
- oxidative stress
- endothelial cells
- cell proliferation
- magnetic nanoparticles
- mass spectrometry
- liquid chromatography
- single cell