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Different types of cartilage neotissue fabricated from collagen hydrogels and mesenchymal stromal cells via SOX9, TGFB1 or BMP2 gene transfer
Author(s) -
Manuel Weißenberger,
Mike Wagenbrenner,
Tizian Heinz,
Jenny Reboredo,
Boris Michael Holzapfel,
Maximilian Rudert,
Jürgen Gröll,
Christopher H. Evans,
Andre F. Steinert
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0237479
Subject(s) - mesenchymal stem cell , self healing hydrogels , bone morphogenetic protein 2 , microbiology and biotechnology , cartilage , sox9 , gene transfer , stromal cell , articular cartilage , chemistry , gene , biology , anatomy , gene expression , medicine , cancer research , biochemistry , pathology , osteoarthritis , alternative medicine , organic chemistry , in vitro
Objective As native cartilage consists of different phenotypical zones, this study aims to fabricate different types of neocartilage constructs from collagen hydrogels and human mesenchymal stromal cells (MSCs) genetically modified to express different chondrogenic factors. Design Human MSCs derived from bone-marrow of osteoarthritis (OA) hips were genetically modified using adenoviral vectors encoding sex-determining region Y-type high-mobility-group-box ( SOX ) 9 , transforming growth factor beta (TGFB) 1 or bone morphogenetic protein ( BMP) 2 cDNA, placed in type I collagen hydrogels and maintained in serum-free chondrogenic media for three weeks. Control constructs contained unmodified MSCs or MSCs expressing GFP. The respective constructs were analyzed histologically, immunohistochemically, biochemically, and by qRT-PCR for chondrogenesis and hypertrophy. Results Chondrogenesis in MSCs was consistently and strongly induced in collagen I hydrogels by the transgenes SOX9 , TGFB1 and BMP2 as evidenced by positive staining for proteoglycans, chondroitin-4-sulfate (CS4) and collagen (COL) type II, increased levels of glycosaminoglycan (GAG) synthesis, and expression of mRNAs associated with chondrogenesis. The control groups were entirely non-chondrogenic. The levels of hypertrophy, as judged by expression of alkaline phosphatase (ALP) and COL X on both the protein and mRNA levels revealed different stages of hypertrophy within the chondrogenic groups ( BMP2 > TGFB1 > SOX9 ). Conclusions Different types of neocartilage with varying levels of hypertrophy could be generated from human MSCs in collagen hydrogels by transfer of genes encoding the chondrogenic factors SOX9 , TGFB1 and BMP2 . This technology may be harnessed for regeneration of specific zones of native cartilage upon damage.

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