Matrix levels of metalloproteinase-2 (MMP-2) and toxicity evaluation of carbonate apatite-based endodontic sealer in rat subcutaneous implantation

Abstract

Objective: The aims of this study is to investigate the matrix metalloproteinase-2 (MMP-2) levels and the toxicity evaluation to determine the safety of carbonate apatite material that will be used as an endodontic sealer. Methods: Carbonate apatite as endodontic sealer material is fabricated with a size of 2 × 2 × 1 mm, sterilized, and then implanted in the subcutaneous area on the murine back. Total of 28 rats was divided into 2 groups: the implantation group and the control group (positive and negative). We observed behavior, macroscopic and microscopic images (through hematoxylin-eosin/HE staining) and MMP-2 levels in the serum (through ELISA examination) to determine the reaction of implant material in experimental animals. Results: Behavioral test shows impaired motor function within 2–24 h in all rats, autonomic behavioral test does not show any disturbance, macroscopic images show appearance of tumor and rubor within 2–72 h in all rats, microscopic images shows an increase in the mean neutrophils number with the highest point located in the 24 h (3.92×103/μL) and lymphocytes with the highest point in the 14 days (15.31×103/μL). The average value of MMP-2 levels of the negative control group was 1.33 ng/ml, 1.29 ng/ml of the positive control group, and 7.32 ng/ml of the implantation group. Level of MMP-2 in rats with visible implants increased on day 3 (0.75 ng/ml), day 10 (1.49 ng/ml) and day 14 (17.67 ng/ml). Conclusion: The implantation of carbonate apatite material did not cause behavioral disorders or abnormalities in the tissues surrounding of the implant site and did not show signs of toxicity or death.

Keywords

• Matrix metalloproteinase-2 (MMP-2)
• Behavioral alteration
• Tissue reaction
• Carbonate apatite
• Endodontic sealer material
• Tissue engineering