Bovine cementum extrac influences murine dental follicle cells in vitro

ABSTRACT

This study evaluated the attachment, chemo-attractive, proliferative and mineralization inductive potential of a bovine cementum extract (CPE) on newborn murine dental follicle cells (MDFC) in vitro. Cementum extract was partially purified by DEAE-cellulose chromatografy. A band representing and Mr of 55,000 was excised form the fel and the protein (s) were electroeluted. Attachment assays revealed that CPE (1.0 µg/ml) promoted MDFC attachment by 96 percent in comparison with collagen type I (5 µg/ml), and was five-fold greater compared with serum-free media (SFM), (p<0.05). Between 1 and 5 days CPE at 1.0 µg/ml and collagen type I at 5 µg/ml sustained more than 75 percent attachement and spreading of MDFC when compared to SFM (P<0.05). Contrary to other reports, fibronectin (0.5 µg/ml) was more potent than CPE in promoting MDFC chemoattraction (P<0.05). MDFC proliferation was stimulated by CPE (0.125 µg/ml), but this response was elicited only when CPE was used together with 10 percent FBS (37.3 percent) or 0.2 percent FBS (76 percent) (p<0.05). Alkaline phosphatase expression by MDFC was increased by CPE (1.0 µg/ml), in comparison to the control. Calcium deposits were detected by von Kossa staining in 14-day MDFC cultures treated with CPE. Nodule formation and its mineralization in long-term MDFC cultures were induced by CPE (1.0 µg/ml). Molecules(s) contained in CPE appear to regulate various biological activities in MDFC, indicating that CPE could play a key role in selecting progenitor cells required for the process of cementogenesis during development