Three‐dimensional cartilage tissue engineering using adult stem cells from osteoarthritis patients

TLDR

The study aims to determine if 3‑D hyaline cartilage can be engineered from bone‑marrow mesenchymal stem cells of osteoarthritis patients, with or without PTHrP to control hypertrophy. BMSCs from OA patients were seeded onto polyglycolic acid scaffolds and differentiated with TGF‑β3, with or without PTHrP, while micromass pellets served as controls; the resulting constructs were then analyzed for mRNA, matrix proteins, and weight. The engineered constructs were five times heavier than pellets, produced abundant proteoglycan and type II collagen with minimal type I, matched bovine nasal chondrocyte protein levels, and, although initially high in type X collagen mRNA, PTHrP treatment suppressed hypertrophy markers while preserving matrix, demonstrating that OA‑derived BMSCs can generate hyaline cartilage suitable for lesion repair.

Abstract

To determine whether it is possible to engineer 3-dimensional hyaline cartilage using mesenchymal stem cells derived from the bone marrow (BMSCs) of patients with osteoarthritis (OA).Expanded BMSCs derived from patients with hip OA were seeded onto polyglycolic acid scaffolds and differentiated using transforming growth factor beta3 in the presence or absence of parathyroid hormone-related protein (PTHrP) to regulate hypertrophy. Micromass pellet cultures were established using the same cells for comparison. At the end of culture, the constructs or pellets were processed for messenger RNA (mRNA) analysis by quantitative real-time reverse transcription-polymerase chain reaction. Matrix proteins were analyzed using specific assays.Cartilage constructs engineered from BMSCs were at least 5 times the weight of equivalent pellet cultures. Histologic, mRNA, and biochemical analyses of the constructs showed extensive synthesis of proteoglycan and type II collagen but only low levels of type I collagen. The protein content was almost identical to that of cartilage engineered from bovine nasal chondrocytes. Analysis of type X collagen mRNA revealed a high level of mRNA in chondrogenic constructs compared with that in undifferentiated BMSCs, indicating an increased risk of hypertrophy in the tissue-engineered cells. However, the inclusion of PTHrP at a dose of 1 microM or 10 microM during the culture period resulted in significant suppression of type X collagen mRNA expression and a significant decrease in alkaline phosphatase activity, without any loss of the cartilage-specific matrix proteins.Three-dimensional hyaline cartilage can be engineered using BMSCs from patients with OA. This method could thus be used for the repair of cartilage lesions.