Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (26): 4101-4108.doi: 10.3969/j.issn.2095-4344.1346
Cobalt-chromium particles inducing preosteoblasts may aggravate periprosthetic inflammation
- 1Department of Traumatic Orthopedics, Binzhou Medical University Hospital, Binzhou 256600, Shandong Province, China; 2Department of Orthopedic Surgery, the University of Kansas School of Medicine-Wichita, Wichita, KS, USA; 3Department of Biological Sciences, Wichita State University, Wichita, KS, USA
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Contact:Yang Shuye, MD, Attending physician, Department of Traumatic Orthopedics, Binzhou Medical University Hopital, Binzhou 256600, Shandong Province, China -
About author:Jiang Jianhao, MD, Attending physician, Department of Traumatic Orthopedics, Binzhou Medical University Hospital, Binzhou 256600, Shandong Province, China -
Supported by:the National Natural Science Foundation of China, No. 81241061 (to ZK); the Medical and Health Technology Development Project of Shandong Province, No. 2016WS0023 (to YSY); the Science Research Startup Foundation of Binzhou Medical University, No. BY2016KYQD19 (to YSY)
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Jiang Jianhao, Li Peng, Du Gangqiang, Liu Hongzhi, Wang Hui, Zhang Kai, Yang Shangyou, Yang Shuye. Cobalt-chromium particles inducing preosteoblasts may aggravate periprosthetic inflammation[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(26): 4101-4108.
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Functionality of osteoblasts induced with Co-Cr particles by ALP activity MC3T3-E1 cells induced with Co-Cr particles exhibited significant negative influence on ALP activity in a dose-dependent manner (P < 0.001). Immunohistochemical staining revealed that only sporadic ALP+ cells were identified in particle-challenged pre-osteoblastic cells, significantly fewer than those in the controls (P < 0.001), which complemented the results of ALP activity assay (Figure 2)."
Gene expression profile changes in MC3T3-E1 cells after Co-Cr particle induction Real-time PCR examination suggested that the MC3T3-E1 cells with Co-Cr particle (1.25 g/L), resulted in significant elevation of chemokine MCP-1 expression (P < 0.001), simultaneously, groups with low concentrations of particles also show significant difference compared with naive cell group. Proinflammatory cytokine genes such as tumor necrosis factor-α, IL-6 and IL-8 were significantly up-regulated by Co-Cr particles. Among groups, Co-Cr particles at 1.25 g/L resulted in highest provocation on IL-6 gene expression (Figure 3)."
Treatment with higher concentrations of the alloy particles (1.25 g/L) p-regulated RANKL gene expression (Figure 3) and down-regulated OPG gene expression (Figure 3), which resulted in much higher RANKL/OPG ratio than controls. NFATc1 gene expression was stimulated by particles in dose-dependent pattern (P < 0.001). Among groups, the Co-Cr particle 1.25 g/L) provoked the most NFATc1 gene expression (P < 0.001) (Figure 3). Trend of OSX and Runx2 gene expression profiles following particles changes were similar, exhibited low concentration (0.3 g/L) did not show influence on gene expressions of OSX and Runx2, although high dose of particles (1.25 g/L) significantly inhibited OSX and Runx2 gene expression (Figure 3; P < 0.001). Effect of Co-Cr particles on inflammatory membrane thickness in the mouse joint prosthesis Little pseudo-membrance formation was noticed only in the hematoxylin-eosin staining peri-implant sections from the stable controls (Figure 4A). Compared to the loosening group, other two groups have significant difference (P < 0.001). It appeared that there was a significantly thicker soft tissue in the particle induction group than in the loosening group (P < 0.001; Figure 4B-D)."
Effect of Co-Cr particles on the BMD in the mouse joint prosthesis The initial micro-CT scan showed that all the metal implants were well placed in position. However, the scans at sacrifice suggested that there were many focal bone resorptions or pit erosions on specimens from Co-Cr particles inducing MC3T3-E1 cell groups compared to the control group. As to the quantification of BMD and BV/TV, the Co-Cr group was significantly different from the other two groups including control group (P < 0.05, P < 0.001). In detail BMD at the peri-implant bone areas among groups indicated a much more severe BMP loss in the challenged-cell transfusion groups, compared to the loosening group (P < 0.001). Meanwhile, the result of BV/TV was similar to the data of BMD (Figure 5)."
Effect of Co-Cr particles on the implant stability in the mouse joint prosthesis When (9.40 ± 0.49) N was the pick force required on the loosening groups, significantly more force was generally needed to pull out pins in stable control group (11.38± 0.65) N, Cr-Co particle-induced MC3T3-E1 cell transfusion group (6.39±0.52) N (P < 0.05) (Figure 6)."
Number of osteoclasts in the mouse joint prosthesis treated with Co-Cr particles TRAP staining was performed to reveal the osteoclasts presence surrounding the pin implantation. The quantity of TRAP+ cells were significant difference in the groups compared to the loosening group (Figure 7; P < 0.01), furthermore Co-Cr groups demonstrated a higher numbers of TRAP+ cells than the loosening group."
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