An Advanced Bioconjugation Strategy for Covalent Tethering of TGFβ3 with Silk Fibroin Matrices and its Implications in the Chondrogenesis Profile of Human BMSCs and Human Chondrocytes: A Paradigm Shift in Cartilage Tissue Engineering

Abstract

The transforming growth factor β class of cytokines plays a significant role in articular cartilage formation from mesenchymal condensation to chondrogenic differentiation. However, their exogenous addition to the chondrogenic media or immobilization onto the scaffolds makes the protocol expensive. Additionally, it reduces the bioavailability of the cytokine to the cells owing to their burst release. Our present study demonstrated an advanced bioconjugation strategy to conjugate TGFβ3 with silk fibroin matrix covalently via a cyanuric chloride coupling reaction. The tethering and change in secondary conformation were confirmed using various spectroscopic analyses. To assess the biological functionality of the chemically modified silk matrix, human BMSCs and chondrocytes were cultured for 28 days in a chondrogenic differentiation medium. Gene expression and histological analysis revealed enhanced expression of chondrogenic markers with intense Safranin‐O and Alcian Blue staining in TGFβ3 conjugated silk matrices than where TGFβ3 was exogenously added to the media for both human BMSCs and chondrocytes. Therefore, this study successfully recapitulated the native niche of TGFβ3 and the role of the silk matrix as a growth factor stabilizer. When cultured over TGFβ3 conjugated silk matrices, human BMSCs displayed increased proteoglycan secretion and maximum chondrogenic trait with attenuation of chondrocyte hypertrophy over human chondrocytes.This article is protected by copyright. All rights reserved

Silk–gelatin hybrid hydrogel: a potential carrier for RNA therapeutics

“…36 When examining the RMSD values, rRNA and tRNA demonstrate more robust interactions with SF, collagen, and chitosan compared to mRNA. 37,38…”