In vitro differentiation of dental pulp stem cells into the chondrogenic route

Abstract

AIM: Cartilage cells have a limited ability to regenerate and self-repair. A large number of people are affected by cartilage deformity due to trauma, degenerative disease, congenital craniofacial defect, idiopathic condylar resorption, ageing, etc. The limited availability of donor sites for cartilage reconstruction makes it an ideal candidate for tissue engineering. Currently bone marrow aspirate is the most popular source of mesenchymal stem cells. Human dental pulp tissue has been found to be an alternative rich source of stem cells. The aim of this study was to examine the in vitro potential of dental pulp stem cells (DPSC) to differentiate into chondrogenic lineage when added to human transforming growth factor beta 3 (TGF-ß3). MATERIALS AND METHOD: A strain of stem cells were isolated from human dental pulp, expanded and further characterized. TGF-ß3 was chosen to trigger its differentiation to cartilage. The TGF-ß3 gene was subcloned as a recombinant pcDNA3.1/V5-His-TOPO-TGF-ß3 vector. The vector was transfected into the DPSC. Three different negative controls were used; blank cells, empty vector and non-related green fluorescent protein (GFP) vector. After 48 hours, the mRNA level of collagen type II and aggrecan was measured by real-time polymerase chain reaction. RESULTS: The relative transcription level of collagen type II mRNA in the TGF-ß3 group was significantly higher than all three negative controls. The relative levels of collagen type II mRNA were 2.746 for blank cell, 154.3 for empty vector, 49.41 for non-related GFP vector and 233.4 for TGF-ß3 vector. CONCLUSION: TGF-ß3 was successful in triggering DPSC differentiation into chondrogenic progenitor cells. It is concluded that DPSC have the capability to form cartilage cells.