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大鼠皮下植入实验中金属蛋白酶-2(MMP-2)的基质水平及碳酸盐磷灰石基根管封闭剂的毒性评估

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

作者信息

Tjahajawati Sri, Zulhazmi Arif, Cahyanto Arief, Mariam Marry Siti, Djustiana Nina, Aripin Dudi, Usri Kosterman, Widyaputra Sebastian Sunardhi

机构信息

Department of Oral Biology, Faculty of Dentistry, Padjadjaran University, Jalan Raya Bandung-Sumedang KM 21, Jatinangor 45363, West Java, Indonesia.

Graduate Student of Faculty of Dentistry, Padjadjaran University, Jalan Sekeloa Selatan 1, Bandung 40134, West Java, Indonesia.

出版信息

Heliyon. 2020 Jul 7;6(7):e04330. doi: 10.1016/j.heliyon.2020.e04330. eCollection 2020 Jul.

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 (3.92×10/μL) and lymphocytes with the highest point in the 14 days (15.31×10/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.

摘要

目的

本研究旨在调查基质金属蛋白酶-2(MMP-2)水平并进行毒性评估,以确定用作牙髓封闭剂的碳酸磷灰石材料的安全性。

方法

制备尺寸为2×2×1mm的碳酸磷灰石作为牙髓封闭剂材料,进行灭菌,然后植入小鼠背部的皮下区域。将28只大鼠分为2组:植入组和对照组(阳性和阴性)。我们观察行为、宏观和微观图像(通过苏木精-伊红/HE染色)以及血清中的MMP-2水平(通过ELISA检测),以确定实验动物中植入材料的反应。

结果

行为测试显示所有大鼠在2-24小时内运动功能受损,自主行为测试未显示任何干扰,宏观图像显示所有大鼠在2-72小时内出现肿瘤和红肿,微观图像显示中性粒细胞平均数量增加,最高点位于24小时(3.92×10/μL),淋巴细胞最高点位于14天(15.31×10/μL)。阴性对照组MMP-2水平的平均值为1.33ng/ml,阳性对照组为1.29ng/ml,植入组为7.32ng/ml。有可见植入物的大鼠中MMP-2水平在第3天(0.75ng/ml)、第10天(1.49ng/ml)和第14天(17.67ng/ml)升高。

结论

碳酸磷灰石材料的植入未引起行为障碍或植入部位周围组织的异常,也未显示出毒性或死亡迹象。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61a1/7347648/93388f8a1b2c/gr1.jpg

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