Characterization of bio-functionalized 316L stainless steel substrates and in vitro study of their early stage influence on osteoblasts behavior - Archive ouverte HAL Accéder directement au contenu
Poster De Conférence Année : 2022

Characterization of bio-functionalized 316L stainless steel substrates and in vitro study of their early stage influence on osteoblasts behavior

Résumé

The implantation of metallic orthopedic prostheses is increasingly common due to the aging of the population and accidents of life. In France alone, orthopaedic prostheses represent an estimated market of 500 million € that continues to grow. But for a part of the world’s population, the access to technically developed prostheses is limited due to expensive care and lack of social support. Thus, there is a real societal need to implement new metal implants that combine durability, good mechanical properties, excellent biocompatibility as well as affordable cost to ensure universal access to medicines and medical devices. The functionalization of low-cost 316L stainless-steel substrates from successive electrodeposition of a polypyrrole film (PPy) and a calcium phosphate deposit doped with silicon was previously carried out at the LPPI and ERRMECe labs1. We are now developing a bio-functionalization of our steel supports coated with a film of PPy by electrodeposition of fibronectin (Fn), a matrix protein involved in adhesion and cellular differentiation. Effects of different modes of electrodeposition or electrooxidation on the structure and functionality of Fn is studied and validated by the cellular behaviour. First electropolymerisation of pyrrole is performed onto steel supports to obtain a primary layer of PPy by cyclic voltammetry. This PPy coating allows passivation of the surface of the steel support and maintain it electrical conductivity. The structure of our PPy coatings is characterized by physico-chemical methods Then Fn is weather adsorbed (ADS), adsorbed and oxidized (OX) or electrodeposited (ED) on PPy coated support. Fn present on our supports is quantified and characterized by enzyme-linked immunosorbent assay (ELISA). Fn organization onto supports is analyzed by confocal microscopy. Early stage behavior of osteoblastic cells (STRO-1A+) cultured on Fn functionalized supports is also studied. A homogenous film of PPy is obtained on our support with a thickness of 4.2µm and a low rugosity (0.8µm). The kinetic of Fn adsorption on support is performed, 10 min is the time necessary to reach the plateau. Fn adsorption isotherm was also study. 10µg/mL appears to be the optimal Fn concentration for our experimentation. Quantification of Fn ADS, OX or ED is performed by ELISA tests. The most Fn quantity was detected after electrodeposition. However, the Fn cell binding domain (CBD) is more accessible on ADS Fn compared to other supports. Those results are consistent with the organisation of Fn observed by confocal microscopy. Ox Fn formed little sticks whereas ED Fn formed long fibre organized like a fractal. At the contrary ADS Fn forms a homogenous coating. Osteoblasts cultured for 3h on each support adhere and spread. The most spread cells are observed on ADS Fn which is in accordance with CBD ELISA. Cell viability was also tested after 3h of culture and no significant toxicity is detected. Our results demonstrated that electrodeposition permits the deposition of Fn higher than a simple adsorption. This technic is fast and can be performed on conductive implant whatever its structure is. Furthermore Fn electrodeposited is not toxic for cells and influences their adhesion and morphology.
Fichier non déposé

Dates et versions

hal-04420638 , version 1 (26-01-2024)

Identifiants

  • HAL Id : hal-04420638 , version 1

Citer

Keerthana Balathandayutham, Olivier Gallet, Séverine Alfonsi, Mathilde Hindié. Characterization of bio-functionalized 316L stainless steel substrates and in vitro study of their early stage influence on osteoblasts behavior. European Society for Biomaterials 2022 (ESB 2022), Sep 2022, Bordeaux, France. ⟨hal-04420638⟩
16 Consultations
0 Téléchargements

Partager

Gmail Facebook X LinkedIn More