Monitoring In Vitro Extracellular Matrix Protein Conformations in the Presence of Biomimetic Bone-Regeneration Scaffolds Using Functionalized Gold-Edge-Coated Triangular Silver Nanoparticles

Abstract

In the cellular environment, high noise levels, such as fluctuations in biochemical reactions, protein variability, molecular diffusion, cell-to-cell contact, and pH, can both mediate and interfere with cellular functions. In this work, gold edge-coated triangular silver nanoparticles (AuTSNP) were validated as a promising new tool to indicate protein conformational transitions in cultured cells and to monitor essential protein activity in the presence of an optimized bone biomimetic chitosan-based scaffold whose rational design mimics the ECM as a natural scaffold. A chitosan-based scaffold formulation with hydroxyapatite (CS/HAp) was selected due to its promising features for orthopedic applications, including combined high mechanical strength biocompatibility and biodegradability. Functionalized AuTSNP-based tests with the model ECM protein, fibronectin (Fn), illustrate that the protein interactions can be clearly sensed over time through the local surface plasmon resonance (LSPR) technique. This demonstrates that AuTNSP are a powerful tool to detect protein conformational activity in the presence of biomimetic bone tissue regeneration scaffolds within a cellular environment that comprises a diversity of molecular cues.

Triangular Silver Nanoparticles Synthesis: Investigating Potential Application in Materials and Biosensing

“…Fn has an uncoiled conformation when in acidic or high salt conditions, whereas within low salt conditions it shows a "coiled compact" conformation [61]. In previous studies where Fn conformational changes are analyzed within different pH environments, Fn is reported to have a hydrodynamic radius of 23 nm when compacted, and when it extends the radius increases up to 130 nm for single Fn strands [43,44]. Fn can form much larger strands as networks and fibrils can develop and be mediated within the extracellular matrix (ECM) [62].…”

“…Growth percentage was calculated using the following equation (Equation ( 1 Nanoplate protection against etching by chloride ions present in physiological environments was achieved by covering the edges of the nanoplates with gold to keep their plasmonic resonance properties. Gold edge coating of the TSNP was carried out following the method reported by [44], consisting of the use of 20 µL of 0.5 mM HAuCl 4 salt and 18.9 µL of 10 mM ascorbic acid per 1 mL of TSNP solution. In this method, there is a reduction in AuCl 4 and inhibition of Ag oxidation at the surface of the TSNP, resulting in the deposition of a thin layer of gold in the edges of the TSNP due to galvanic replacement.…”

“…Brennan-Fournet et al (2015) successfully detected conformational changes in fibronectin, a protein related to tumor growth and cell survival, using AuTSNPs [43]. Rodriguez Barroso et al (2023) monitored fibronectin behavior overtime within different complex cellular environments [44]. Vinayagam et al (2018) built a nanoprobe by conjugating single-strand DNA to the TSNP surface, for specific detection of dengue virus [45].…”

A Review of Children’s Prosthetic Self-hygiene Using Antibacterial Silver Nanoparticles (AgNPs)

“…Their unique properties and broad-spectrum antimicrobial activity make them valuable in healthcare applications, including the self-hygiene of sports prostheses for children. By incorporating AgNPs into prosthetic materials, a hygienic environment can be promoted, reducing the risk of infections and enhancing the overall well-being (Akhavan et al, 2018;Sarraf et al, 2018;Vale et al, 2019;Xie et al, 2019;Devaraj et al, 2020;Sohi et al, 2021;Bee et al, 2022;Nathanael et al, 2022;Padmanabhan et al, 2022;Rodriguez Barroso et al, 2023).…”

Triangular Silver Nanoplates as a Bioanalytical Tool: Potential COVID-19 Detection

“…The currently available techniques for protein characterization that are able to detect conformations and structural changes in proteins require expensive instrumentation and extensive data analysis, and their signals are obstructed by cellular noise when applied in complex biological environments [9]. The necessity for the development of enzyme-free analytical assays has arisen in order to overcome ELISA assays' limitations [10].…”