Abstract
Collagen is essential for cartilage adhesion and formation. In the present study, histology, immunofluorescence, morphometry, and qRT-PCR suggested that adipose-derived stem cells (ADSCs) stimulated by type V collagen (Col V) induce a significant increase of type II collagen (Col II) in the degenerative area of surgical-induced osteoarthritic rabbit articular cartilage (OA). In vitro, the effects of Col V on the proliferation and differentiation of ADSC were investigated. The expression of the cartilage-related genes Col2a1 and Acan was significantly upregulated and Pou5fl was downregulated post-ADSC/Col V treatment. Post-ADSC/Col V treatment, in vivo analyses revealed that rabbits showed typical signs of osteoarthritic articular cartilage regeneration by hematoxylin and eosin (H&E) and Safranin O/Fast Green staining. Immunohistochemical staining demonstrated that the volume of Col II fibers and the expression of Col II protein were significantly increased, and apoptosis Fas ligand positive significantly decreased post-ADSC/Col V treatment. In conclusion, the expression of Col II was higher in rabbits with surgical-induced osteoarthritic articular cartilage; hence, ADSC/Col V may be a promising therapeutic target for OA treatment.
Highlights
MATERIALS AND METHODSIn humans, osteoarthritis (OA), which involves the degradation of joint cartilage and underlying bone, makes the collagen matrix disorganize and decreases the proteoglycan content within the cartilage (Sanchez-Adams et al, 2014)
adipose-derived stem cells (ADSCs) grown in specific differentiation media were able to differentiate into adipogenic, chondrogenic, and osteogenic lineages, characterized, respectively, by the presence of cytoplasmic lipid droplets (Oil Red), synthesis of proteoglycans by the presence of a proteoglycan-rich matrix (Alcian Blue), and aggregates of calcium salts with calcium deposits (Alizarin Red) (Figures 2A–D)
The current study showed that ADSCs stimulated by Col V (ADSCs/Col V) induced a significant regeneration of cartilage, indicating that ADSCs/Col V may be a therapeutic target for the treatment of osteoarthritis
Summary
Osteoarthritis (OA), which involves the degradation of joint cartilage and underlying bone, makes the collagen matrix disorganize and decreases the proteoglycan content within the cartilage (Sanchez-Adams et al, 2014). The present study evaluated the expression profiles of Col II in rabbits with surgical-induced osteoarthritic articular cartilage treated with adipose-derived stem cells (ADSCs) stimulated by Col V. The cell cultures were maintained for a period of 1, 2, 3, 7, 14, and 21 days These cells were cultured over coverslips in DMEM (Gibco Life Technologies, Invitrogen, Paisley, United Kingdom) supplemented with 10% fetal bovine serum (GibcoTM) and 1% penicillin/streptomycin (Sigma Chemical Co., St. Louis, MO, United States) and were. After Col V stimuli evaluation, ADSCs in the third culture passage were cultivated with Col V (50 μg/ml) per 72 h in 75 cm culture flasks with DMEM (Gibco Life Technologies, Invitrogen, Paisley, United Kingdom), supplemented with 10% fetal bovine serum (GibcoTM) and 1% penicillin/streptomycin (Sigma Chemical Co., St. Louis, MO, United States), at 37◦C in an atmosphere of 5% CO2. Results are expressed as mean ± standard errors of the mean (SEM); a P < 0.05 was considered statistically significant
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