Supplementation of articular cartilage-derived chondroprogenitors with bone morphogenic protein-9 enhances chondrogenesis without affecting hypertrophy

• Published in: Biotechnology letters 2022-09, Vol.44 (9), p.1037-1049
• Main Authors: Padmaja, Kawin, Amirtham, Soosai Manickam, Rebekah, Grace, Sathishkumar, Solomon, Vinod, Elizabeth
• Format: Article
• Language: English
• Subjects: Applied Microbiology; Biochemistry; Biomarkers; Biomedical and Life Sciences; Biotechnology; Bone morphogenetic proteins; Cartilage; Cartilage (articular); Cartilage diseases; Cell culture; Cell cycle; Chondrocytes; Chondrogenesis; Culture; Differentiation; Eutrophication; Fibronectin; Gene expression; Glycosaminoglycans; Growth factors; Growth kinetics; Hypertrophy; Life Sciences; Mesenchyme; Microbiology; Original Research Paper; Osteoarthritis; Parathyroid hormone; Proteins; Qualitative analysis; Stem cells; Supplements; Surface markers; Toluidine; Toluidine blue
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-022-03280-9

Introduction Chondroprogenitors (CPCs) have emerged as a promising cellular therapy for cartilage-related pathologies due to their inherent primed chondrogenic potential. Studies report that the addition of growth factors such as parathyroid hormone (PTH) and Bone Morphogenic Protein (BMP) enhance the chondroinducive potential in chondrocytes and mesenchymal stem cells. This study evaluated if supplementation of the standard culture medium for cell expansion with 1–34 PTH and BMP-9 would enhance the chondrogenic potential of CPCs and reduce their hypertrophic tendency. Methods Human chondrocytes were isolated from patients undergoing total knee replacement for osteoarthritis (n = 3). Following fibronectin adhesion assay, passage 1 CPCs were divided and further expanded under three culture conditions (a) control, i.e., cells continued under standard culture conditions, (b) 1–34 PTH group, additional intermittent 6 h exposure with 1–34 PTH and (c) BMP-9 group, additional BMP-9 during culture expansion. All the groups were evaluated for population-doubling, cell cycle analysis, surface marker and gene expression for chondrogenesis, hypertrophy, multilineage differentiation and GAG (glycosaminoglycan)/DNA following chondrogenic differentiation. Results Concerning growth kinetics, the BMP-9 group exhibited a significantly lower S-phase and population-doubling when compared to the other two groups. Qualitative analysis for chondrogenic potential (Alcian blue, Safranin O staining and Toluidine blue for GAG) revealed that the BMP-9 group exhibited the highest uptake. The BMP-9 group also showed significantly higher COL2A1 expression than the control group, with no change in the hypertrophy marker expression. Conclusion BMP-9 can potentially be used as an additive for CPCs expansion, to enhance their chondrogenic potential without affecting their low hypertrophic tendency. The mitigating effects of 1-34PTH on hypertrophy would benefit further investigation when used in combination with BMP-9 to enhance chondrogenesis whilst reducing hypertrophy.