title: Biosynthesis of collagen I, II, RUNX2 and lubricin at different time points of chondrogenic differentiation in a 3D in vitro model of human mesenchymal stem cells derived from adipose tissue.
abstract: The first aim of the study was to identify the most appropriate time for differentiation of adipose tissue derived mesenchymal stem cells (MSCs) to chondrocytes, through the self-assembly process. For this purpose, the expression of some chondrocyte markers, such as collagen type I, collagen type II, RUNX2 and lubricin was investigated at different times (7, 14, 21 and 28 days) of chondrogenic differentiation of MSCs, by using immunohistochemistry and Western blot analysis. The second aim of the study was to demonstrate that the expression of lubricin, such as the expression of collagen type II, could be a possible biomarker for the detection of chondrocytes well-being and viability in the natural self-assembling constructs, called 'cell pellets'. Histology (hematoxylin and eosin) and histochemistry (alcian blue staining) methods were used to assess the chondrogenic differentiation of MSCs. The results showed that after 21 days the differentiated chondrocytes, when compared with MSCs cultured without chondrogenic medium (CD44, CD90 and CD105 positive; CD45, CD14 and CD34 negative), were able to produce significant quantities of collagen type I, collagen type II, and lubricin, suggesting hyaline cartilage formation. During the differentiation phase, the cells showed a reduced expression of RUNX2, a protein expressed by osteoblasts. Our studies demonstrated that 21 days is the optimum time for the implantation of chondrocytes differentiated from adipose tissue-derived MSCs. This information could be useful for the future development of cell-based repair therapies for degenerative diseases of articular cartilage.
authors.affiliation: Department of Bio-Medical Sciences, Human Anatomy and Histology Section, School of Medicine, University of Catania, Catania, Italy. Electronic address: g.musumeci@unict.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Musumeci
authors.affiliation: Department of Veterinary Preclinical Sciences, School of Veterinary Medicine, Faculty of Health and Medical Sciences, University of Surrey, Guildford, United Kingdom; Arthritis Research UK Centre for Sport, Exercise and Osteoarthritis, Arthritis Research UK Pain Centre, Medical Research Council and Arthritis Research UK Centre for Musculoskeletal Ageing Research, University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom; Center of Excellence in Genomic Medicine Research (CEGMR), King Fahd Medical Research Center (KFMRC), King AbdulAziz University, Jeddah, Kingdom of Saudi Arabia.
authors.forename: Ali
authors.initials: A
authors.lastname: Mobasheri
authors.affiliation: Department of Medical and Pediatric Sciences, Internal Medicine Division, University of Catania, Catania, Italy.
authors.forename: Francesca Maria
authors.initials: FM
authors.lastname: Trovato
authors.affiliation: Department of Bio-Medical Sciences, Human Anatomy and Histology Section, School of Medicine, University of Catania, Catania, Italy.
authors.forename: Marta Anna
authors.initials: MA
authors.lastname: Szychlinska
authors.affiliation: Department of Bio-Medical Sciences, Section of Physiology, University of Catania, Catania, Italy.
authors.forename: Adriana Carol Eleonora
authors.initials: AC
authors.lastname: Graziano
authors.affiliation: Department of Bio-Medical Sciences, Section of Physiology, University of Catania, Catania, Italy.
authors.forename: Debora
authors.initials: D
authors.lastname: Lo Furno
authors.affiliation: Department of Bio-Medical Sciences, Section of Physiology, University of Catania, Catania, Italy.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Avola
authors.affiliation: Department of Surgery, Orthopaedics and Traumatology Division, University of Catania, Catania, Italy.
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Mangano
authors.affiliation: Department of Bio-Medical Sciences, Section of Physiology, University of Catania, Catania, Italy.
authors.forename: Rosario
authors.initials: R
authors.lastname: Giuffrida
authors.affiliation: Department of Bio-Medical Sciences, Section of Physiology, University of Catania, Catania, Italy.
authors.forename: Venera
authors.initials: V
authors.lastname: Cardile
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D020219:Chondrogenesis / Q000502:physiology; D024042:Collagen Type I / Q000378:metabolism*; D024043:Collagen Type II / Q000378:metabolism*; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D000099158:Proteoglycan 4
chemical_list: D024042:Collagen Type I; D024043:Collagen Type II; D050659:Core Binding Factor Alpha 1 Subunit; D006023:Glycoproteins; D000099158:Proteoglycan 4
keywords: Adipose tissue; Chondrocytes; Collagen; Lubricin; Mesenchymal stem cells; RUNX2; Regeneration