Overexpression of DMP1 in Periodontal Ligament Stem Cells using Lentivirus

Objectives: Periodontitis and the resultant inflammation can lead to destruction of connective tissue attachment to the tooth, resorption of alveolar bone and ultimately tooth loss. Periodontal ligament contains the necessary stem cells or progenitor cells that have the ability to regenerate lost periodontal tissues. Expression of DMP1 is vital in tooth and bone mineralization, however, little is known about the function of DMP1 in PDL cells. Therefore, in this study we intend to develop a lentiviral based DMP1 overexpressing PDL cells and characterize their phenotype as either osteoblasts, cementoblasts or odontoblasts. Methods: pLenti-EF1a-GFP-2A Puro Vector was used to clone DMP1 cDNA according to the manufacturer's protocol. Experimental conditions are being standardized for optimizing transduction conditions for human PDLs and characterization of the resultant DMP1 overexpressing PDLs. Results: Efficient transduction of DMP1containing lentiviral vector was obtained in 293T cells as ascertained by the expression of the green fluorescent protein contained in the vector. Human PDLs are currently being transduced using an optimized multiplicity of infection. Conclusions: Several experimental conditions are being standardized for improved production and transduction conditions for lentiviral vectors containing DMP1. These improvements should facilitate in establishing a stable PDL cell-line overexpressing DMP1 for further characterization.