Isolation and characterization of lineages cementocytes

Abstract

The cementocytes not only share morphological features with osteocytes, but also express several metabolism regulators of hard tissue in common. However knowledge on cellular and molecular level about cementocytes and their role in the homeostasis of dental cementum remain unclear. Thus, the objective of this study is to obtain and characterize a cementocyte cell line obtained from human teeth without caries or periodontal disease and, wicht presente complete formation of dental root. These teeth were extracted due to orthodontic or prosthetic reasons. Right after teeth extracting will be placed in a biopsy. Then, over a sterile petri dish (35x100mm), the periodontal ligament is carefully removed from the entire length of the root, and using a Gracey 5/6 curette and a clinical microscope. The dental cement matrix along with the apical cementocytes (cementocytes area of greatest density) will be carefully removed from the root surfasse. These fragments transferred to a cell culture dish (35x100mm) and kept in a biopsy for 24 hours 37°C in a saturated atmosphere at 5% CO2 and 98% humidity. The media of the biopsy will be replaced by standard culture and when they develop confluence, cells will be trypsinized and frozen in liquid nitrogen for future experiments. There will be obteined cell clones and will be characterized using gene expression and functional assays. From the results of this proposal, it is expected to develop an in vitro model obtained from human tissues to study the contribution of cementocytes on the control the homeostasis of the dental cement, identifying biomolecules with potential use on the controlling the formation and regeneration of tissue. (AU)