Adipose-Derived Mesenchymal Stem Cell Chondrospheroids Cultured in Hypoxia and a 3D Porous Chitosan/Chitin Nanocrystal Scaffold as a Platform for Cartilage Tissue Engineering

Abstract

Articular cartilage degeneration is one of the most common causes of pain and disability in middle-aged and older people. Tissue engineering (TE) has shown great therapeutic promise for this condition. The design of cartilage regeneration constructs must take into account the specific characteristics of the cartilaginous matrix, as well as the avascular nature of cartilage and its cells’ peculiar arrangement in isogenic groups. Keeping these factors in mind, we have designed a 3D porous scaffold based on genipin-crosslinked chitosan/chitin nanocrystals for spheroid chondral differentiation of human adipose tissue-derived mesenchymal stem cells (hASCs) induced in hypoxic conditions. First, we demonstrated that, under low oxygen conditions, the chondrospheroids obtained express cartilage-specific markers including collagen type II (COL2A1) and aggrecan, lacking expression of osteogenic differentiation marker collagen type I (COL1A2). These results were associated with an increased expression of hypoxia-inducible factor 1α, which positively directs COL2A1 and aggrecan expression. Finally, we determined the most suitable chondrogenic differentiation pattern when hASC spheroids were seeded in the 3D porous scaffold under hypoxia and obtained a chondral extracellular matrix with a high sulphated glycosaminoglycan content, which is characteristic of articular cartilage. These findings highlight the potential use of such templates in cartilage tissue engineering.

Marine-Derived Polymeric Materials and Biomimetics: An Overview

“…In 2001 Paillet and Dufresne used chitin nanocrystals for the first time as ecofriendly reinforcing fillers in thermoplastic nanocomposites [145]. In the following years, new domains of exploitation of chitin nanofillers have emerged including bionanocomposite materials, electronics and medical devices [43,[146][147][148][149][150][151]].…”

“…Zubillaga et al developed films and 3D porous scaffolds chitosan-genipin/chitin nanofillers as platforms for cartilage tissue engineering using adipose-derived mesenchymal stem cell chondrospheroids cultured in hypoxia [43,151].…”

Extraction of Nanochitin from Marine Resources and Fabrication of Polymer Nanocomposites: Recent Advances

“…Based on these approaches, whiskers have recently been prepared from chitins of different sources, such as crab shells, shrimp shells, lobster shells and squid pens [29][30][31][32][33][34][35][36].…”

“…Studies were done on the addition of chitin nanocrystals as a compatibilizer in a blend system. Additionally, nanochitin has been blended with hydrophilic and bio-based polymeric matrices, namely, chitosan, starch, PLA, cellulose nanocrystals [33,41,[56][57][58][59]. There are many potential applications for polymer/nanochitin in membrane technology, dye removal, packaging materials, drug delivery, tissue engineering and biochemical relevance.…”

“…Moreover, the addition of chitin nanoforms improved cell adhesion at low chitin nanoform ratios [56]. More recently, the same authors assessed the adipose-derived mesenchymal stem cell chondrospheroids cultured in hypoxia in the same 3D porous scaffolds as a platform for cartilage tissue engineering [33].…”