Bovine cementum extract influences murine dental follicle cells in vitro

This study evaluated the attachment, chemoattractive, 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 chromatography. A band representing an M(r) of 55,000 was excised from the gel and the protein(s) were electroeluted. Attachment assays revealed that CPE (1.0 microgram/ml) promoted MDFC attachment by 96% in comparison with collagen type I (5 micrograms/ml), and was five-fold greater compared with serum-free media (SFM), (P < 0.05). Between 1 and 5 days CPE at 1.0 microgram/ml and collagen type I at 5 micrograms/ml sustained more than 75% attachment and spreading of MDFC when compared to SFM (P < 0.05). Contrary to other reports, fibronectin (0.5 microgram/ml) was more potent than CPE in promoting MDFC chemoattraction (P < 0.05). MDFC proliferation was stimulated by CPE (0.125 microgram/ml), but this response was elicited only when CPE was used together with 10% FBS (37.3%) or 0.2% FBS (76%) (P < 0.05). Alkaline phosphatase expression by MDFC was increased by CPE (1.0 microgram/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 microgram/ml). Molecule(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.