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CJAP ›› 2018, Vol. 34 ›› Issue (1): 83-87.doi: 10.12047/j.cjap.5608.2018.021

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TCP wear particles causes injury of periprosthetic osteocytes in the mouse calvaria

YAN Jia-qi1,2, ZHANG Yun1, LIU Fang-shu1, CAI Ting-ting1, TONG Kang-qiang1, ZHU Can1, HU Lu-qi1, LV Shu-min1   

  1. 1. Department of Basic Medicine, College of Medicine, Shaoxing University, Shaoxing 312000;
    2. Department of Pediatrics, Hangzhou Bay Hospital of Ningbo, Ningbo 315336, China
  • Received:2017-05-31 Revised:2017-10-27 Online:2018-01-28 Published:2018-06-19
  • Supported by:
    浙江省自然科学基金资助项目(LY13H060003,LY17H060007);浙江省重大科技专项重点社会发展项目(2014C03017)

Abstract: Objective: To study whether tricalcium phosphate(TCP) wear particles cause injuries of periprosthetic osteocytes in the mouse calvaria, and to explain its molecular mechanism.Methods: Thirty six-week(ICR)male mice were randomly divided into sham group, model (TCP) group and 3-methyladenine (3-MA) group. A murine calvarial model of osteolysis was established by 30 mg of TCP wear particles implantation over the periosteum around the middle suture of calvaria in mice. On the second postoperative day, the autophagy specific inhibitor 3-MA (1.0 mg/kg) was subcutaneously injected to the calvaria in the 3-MA-treated mice every other day. After 2 weeks, blood and the calvaria were obtained. Micro-CT was used to detect bone mineral density(BMD), bone volume fraction (BVF) and porosity number. HE staining and flow cytometry were performed to analyze the viability and apoptosis of periprosthetic osteocytes. The serum levels of dentin matrix protein 1(DMP-1) and sclerostin (SOST) were determined by ELISA. The proteins expressions of DMP-1, SOST, Beclin-1 and microtuble-associated protein 1 light chain 3 (LC-3) were detected by Western blot in the calvaria osteocytes.Results: Compared with the sham group, the mice in the TCP group showed that a significant decrease in the viability of periprosthetic osteocytes, but obvious increases in number of osteocytes death and osteocytes apoptosis (P<0.05), and in serum level and protein expression of SOST; significant decreases in serum level and protein expression of DMP-1 (P<0.05), and remarkable up-regulation of autophagy-related factors beclin-1 and the conversion of LC3-Ⅱ from LC3-I in the calvaria osteocytes. Compared with TCP group, the mice in the 3-MA group showed that injuries of calvaria osteocytes were obviously aggravated, and osteocytes apoptosis was significantly increased (P<0.05).Conclusion: TCP wear particles can cause injuries of periprosthetic osteocytes via activation of apoptosis and autophagy, which promotes osteolysis around the prosthesis osteolysis and joint aseptic loosening.

Key words: TCP wear particles, mouse, osteocytes, apoptosis, autophagy

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