Title: 1118 - CD146 as a Receptor for CTGF in PDL Stem Cells
Ashleigh Alex (Presenter)
Columbia University College of Dental Medicine
Rebecca Yu, Columbia University College of Dental Medicine
Solaiman Tarafder, Columbia University College of Dental Medicine
Chang Lee, Columbia University College of Dental Medicine
Objectives: We have recently identified a population of CD146+ periodontal ligament stem cells (PDLSCs) that undergo differentiation leading to tissue regeneration. An interesting finding from our previous studies is that connective tissue growth factor (CTGF) selectively stimulates differentiation of PDLSCs while showing minimal effect on CD146- PDL cells. Given the strong surface expression of CD146 as a clear distinction from the other PDL cells, this study was designed to test the hypothesis that CD146 serves as a receptor for CTGF for differentiation of CD146+ PDLSCs.
Methods: A well-established ELISA-based binding assay was adopted and modified to test the binding of CTGF to Fc-CD146. The function of CD146 as a CTGF receptor was investigated by blocking CD146 with antibody in the course of CTGF-induced differentiation of PDLSCs. Briefly, P2 – P3 CD146+ PDLSCs were seeded at 80% confluence in 6-well plates, and CD146 antibody (5 µg/well) was applied to wells. After 2 hour incubation and washing away of the antibody solution, 100 ng/ml CTGF was applied. At 1 wk, qRT-PCR was performed for PDL-related markers including COL-I, COL-III, fibronectin (FN), and tenascin-C (Tn-C) using commercially available primers as per our well-established methods.
Results: Our ELISA-based binding assay showed the OD measures increasing as the CTGF dose increased with a saturation curve, suggesting the binding of Fc-CD146 and CTGF. At 1 wk, blocking CD146 resulted in no noticeable change in cellular phenotype and qRT-PCR analysis demonstrated significantly reduced mRNA expressions of COL-I, COL-III, FN, and TN-C in PDLSCs treated with CTGF (n = 3 per group; p<0.05).
Conclusions: Our data with an identification of CTGF receptors in PDLSCs may have implications in biology of tendon stem/progenitor cells and PDL regeneration.
This abstract is based on research that was funded entirely or partially by an outside source:
Columbia Dental Medicine Summer Research Fellowship 2017
The submitter must disclose the names of the organizations with which any author have a relationship, the nature of the relationship, and the clinical or research area involved. The following is submitted: NONE