In silico and experimental analysis of the interaction between a biomimetic motif of fibronectin and type I collagen: Study of a fibronectin chimera
Lucy Charbit1. Camille Fauré1. Maëla Gars1. Emma Lemeunier1. Emma Richard1. Amina Ben Abla1. Jad Eid1.
1Ecole de Biologie Industrielle. EBInnov. 49 avenue des Genottes. 95895 Cergy-Pontoise. France
ABSTRACT: In the context of regenerative medicine, precise control of cell behaviour is pivotal for successful tissue regeneration. Clinical strategies frequently involve combining biomaterials as scaffolds with bioactive molecules [1] , such as fibronectin, to guide this cellular behaviour.
Biomaterials mimic the Extracellular Matrix (ECM), providing a platform for cell adhesion, growth, and differentiation along with porosity facilitating nutrient transport and cell migration [2], [3]. Fibronectin (FN), a crucial protein linking cells to the ECM, primarily composed of collagen, plays a central role in this modulation.
The presence of FN at wound sites forms a dense tridimensionality network, guiding cell behaviour, proliferation, migration, and contributing to wound contraction. Furthermore, a recent study highlights the enhancement of cellular responses when using fibronectin with biomaterials [4].
Recombinant technology is increasingly used in regenerative medicine to generate customized gene sequences leading to recombinant proteins that are used in different fields. Studies have demonstrated that recombinant chimeric fibronectin proteins can significantly improve cellular healing activity, compared to full-length native proteins [5], [6].
The study aims to assess the efficiency of a bio-adhesive recombinant FN chimeric fragment (r-FNC) when binding collagen. This r-FNC was created in our laboratory and contains FN cell binding domain in fusion with FN collagen binding domain. This recombinant chimera possesses only the specific binding sites, making it a significantly smaller protein compared to native fibronectin. This characteristic enables more efficient coating on biomaterials in regenerative medicine and enhances affinity during interactions.
Several bioinformatics tools were employed to select the most accurate conformation of the designed chimera. The best conformation was identified through structural alignment and stability assessments. Subsequent molecular docking simulations, utilizing both blind and targeted approaches, were conducted to predict the interaction between the r-FNC chimera and collagen. Blind docking explored all possible interactions across the entire surface of the chimera, while targeted docking focused on the specific collagen binding site. The results from both approaches were consistent, demonstrating a strong affinity between r-FNC and collagen. Both in silico and in vitro analyses confirmed the robust affinity and interaction between r-FNC and collagen. The in vitro study highlighted the optimal conditions for this interaction, showcasing the potential of r-FNC in enhancing cellular adhesion and promoting tissue regeneration.
In summary, the groundbreaking nature of this study underscores the critical role of our recombinant proteins r-FNC, in advancing the field of regenerative medicine. These findings represent a significant leap forward, offering invaluable insights that could revolutionize tissue engineering and wound healing therapies. The potential efficacy of r-FNC opens new avenues for the development of biomaterials tailored to promote tissue regeneration, marking a pivotal milestone in the quest for transformative regenerative solutions.
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