Genetic Engineering of Mesenchymal Stem Cells for Differential Matrix Deposition on 3D Woven Scaffolds

Tissue Engineering. Part a
Nguyen P T HuynhFarshid Guilak

Abstract

Tissue engineering approaches for the repair of osteochondral defects using biomaterial scaffolds and stem cells have remained challenging due to the inherent complexities of inducing cartilage-like matrix and bone-like matrix within the same local environment. Members of the transforming growth factor β (TGFβ) family have been extensively utilized in the engineering of skeletal tissues, but have distinct effects on chondrogenic and osteogenic differentiation of progenitor cells. The goal of this study was to develop a method to direct human bone marrow-derived mesenchymal stem cells (MSCs) to deposit either mineralized matrix or a cartilaginous matrix rich in glycosaminoglycan and type II collagen within the same biochemical environment. This differential induction was performed by culturing cells on engineered three-dimensionally woven poly(ɛ-caprolactone) (PCL) scaffolds in a chondrogenic environment for cartilage-like matrix production while inhibiting TGFβ3 signaling through Mothers against DPP homolog 3 (SMAD3) knockdown, in combination with overexpressing RUNX2, to achieve mineralization. The highest levels of mineral deposition and alkaline phosphatase activity were observed on scaffolds with genetically engineered MSCs...Continue Reading

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Citations

Dec 10, 2019·Npj Regenerative Medicine·Mark F PittengerArnold I Caplan
Nov 24, 2021·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Farshid GuilakFranklin T Moutos

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Methods Mentioned

BETA
fluorescence microscopy
phosphatase assay

Software Mentioned

GraphPad
GPP
Graphpad Prism

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