Regenerative potential of mesenchymal multipotent progenitor cells from human periodontal ligament

Abstract

With the main target to promote the regeneration of periodontal tissue destroyed by periodontal disease, using mesenchymal stem cells from periodontal ligament, after it has isolated mesenchymal multipotent progenitor cells from ligament (PDLMSCs) harvested from permanent and primary human teeth, and comparatively assess their stem cell properties, including proliferation rate and viability, multipotency, and the expression of stem-cell marker, the aim of this study will be to compare these two groups of multipotent postnatal stem cells considering: a) potential of differentiation in vitro, b) cellular responses to the growth factors present at the formation and regeneration of periodontal tissues (in vitro), and c) potential of these cells in regenerating periodontal tissues (in vivo). After culturing of PDLMSCs (CD105+ CD34- CD45- cells), and phenotipic characterization in relation to expression of stem cell markers (CD105, CD166, Oct4 and STRO-1), these cells will be used in order to evaluate their osteoblastic/cementoblastic, adipogenic, and chondrogenic phenotype markers, their potential to produce calcified nodules as well as telomerase activity in the two groups of cells. CD105+ CD34- CD45- cells will then be treated with BMP-2 and FGF-8, and the expression of the gene involved at the tooth tissue formation and periodontal tissue regeneration will be evaluated (PCRq). In vivo, cells will be transplanted subcutaneously into the dorsal surfaces in immunocompromised mice and into periodontal defects in immunodeficient rats to evaluate the formation of ectopic mineralized tissue and the cell potential to regenerate the periodontium tissues, respectively, using hystomorphometric analysis. Besides, cells transplanted into the periodontal defects will be captured by Laser Capture Microdissection for molecular analysis of genes involved at the cellular differentiation. This will be carried out using the techniques of Microarray and PCRq. (AU)