Homogenized Porcine Extracellular Matrix Derived Injectable Tissue Construct with Gold Nanoparticles for Musculoskeletal Tissue Engineering Applications

Abstract

A unique porcine extracellular matrix (ECM) derived injectable tissue construct with 100 nm or 20 nm gold nanoparticles (AuNP) was developed for musculoskeletal tissue engineering applications. ECM has been shown to encourage cellularity and tissue remodeling due to its release of growth factors while AuNP have been shown to reduce reactive oxygen species (ROS) levels. Injectable tissue constructs were created by homogenizing decellularized porcine diaphragm tendon conjugated with 100 nm or 20 nm AuNP at 1x, 4x, and 8x concentrations. Extrusion force testing demonstrated that homogenized tissue constructs were injectable at an appropriate cannula size and force. L-929 murine fibroblasts were used to measure cell viability, cell proliferation, intracellular ROS levels, and cell migration in response to constructs. Enhanced cell viability and proliferation are observed on 1 × 20 nm AuNP constructs. ROS assays demonstrate reduced cellular ROS concentrations from all 20 nm AuNP constructs and from 8 × 100 nm AuNP constructs compared with constructs without nanoparticles. Cellular migration is higher towards 4 × 20 nm AuNP constructs compared with constructs without nanoparticles. Results support the potential use of a porcine ECM derived injectable tissue construct with AuNP as an injectable tissue construct to reduce inflammation and to promote tissue remodeling in musculoskeletal tissue engineering applications.

A Comprehensive Review on Bio-Nanomaterials for Medical Implants and Feasibility Studies on Fabrication of Such Implants by Additive Manufacturing Technique

“…The use of metallic nanofillers (e.g., gold or silver nanoparticles) is interesting for biomedical applications to enhance the electrical conductivity or antimicrobial properties of tissues. For example, the incorporation of gold nanoparticles into extracellular matrixes (ECM) demonstrated the enhancement of the cellularity by reinforcing the ECM, while mitigating the inflammation for musculoskeletal tissue engineering applications [24]. Silver, in the form of nanoparticles, is also well recognized for its antimicrobial properties, specifically as an alternative to antibiotic drugs.…”

“…In addition, conjugation, or cross-linking is generally used to assure covalent grafting between nanoparticles and tissues and thus improving the integration of nanofillers within matrixes. For example, Smith et al conjugated gold nanoparticles with a porcine extracellular matrix by first grafting 2-mercaptoethy-lamine (MEA) on gold nanoparticles before conjugation with 1-ethyl-3-[3-dimethylainopropyl] carbodiimide (EDC) [24].…”

Incorporation of nanoparticles into transplantable decellularized matrices: Applications and challenges

“…[23][24][25] Amine functionalization of NPs is performed by conjugation with an organic compound containing both an amine and thiol functional groups such as glutathione, cysteine, or cysteamine; with compounds containing amine and hydroxyl groups such as ethanolamine; or with other organic coating agents, such as butylamine, ethylenediamine, heptylamine, di-amino-propane, allylamine, or ammonia, which provide NPs with amine groups that can react with carboxyl groups in matrices. [26][27][28][29] Carboxyl functionalization of NPs can also be performed using acids that provide reactive carboxyl groups, such as poly(D, L-lactide-co-glycolide) (PLGA) and thioctic acid. 30,31 Although functionalized NPs can more efficiently bind decellularized matrices, their homogeneous distribution within the matrix is altered; this is because functionalized NPs are more likely to aggregate before loading into the matrices.…”

“…That study revealed that AuNPs (especially 20 nm AuNPs) can enhance fibroblast migration and functionality of the injectable extracellular matrix (ECM) constructs. 28 However, although the gold nanostructures generally improve porcine tendinous diaphragmatic material, their size and shape for optimal biocompatibility have not yet been determined. Application of single-walled carbon nanotubes (SWCNTs; 1.5 nm × 200-300 nm), which have excellent mechanical and conductive properties, was examined by Deeken et al 55 That study aimed to improve resistance to collagenase and tensile strength of the diaphragmatic porcine tendon.…”

“…That study revealed that AuNPs (especially 20 nm AuNPs) can enhance fibroblast migration and functionality of the injectable extracellular matrix (ECM) constructs. 28 However, although the gold nanostructures generally improve porcine tendinous diaphragmatic material, their size and shape for optimal biocompatibility have not yet been determined.…”