Influence of Ceramic Debris on Osteoblast Behaviors: AnIn VivoStudy

Guo‐Jing Sun,Yun‐Fan Ti,Shao‐Gang Xu,Jian‐Ning Zhao,Shu‐Feng Yang,Guo‐Dong Guo,Geng‐Tao Fan,Feng‐Rong Chen

doi:10.1111/os.12496

Abstract

ObjectiveWear‐induced aseptic loosening has been accepted as one of the main reasons for failure of total hip arthroplasty. Ceramic wear debris is generated following prosthesis implantation and plays an important part in the upregulation of inflammatory factors in total hip arthroplasty. The present study investigates the influence of ceramic debris on osteoblasts and inflammatory factors.MethodsCeramic debris was prepared by mechanical grinding of an aluminum femoral head and added to cultures of MC3T3‐E subclone 14 cells at different concentrations (i.e. 0, 5, 10, and 15 μg/mL). Cell proliferation was evaluated using a Cell Counting Kit (CCK‐8), and cell differentiation was assessed by mRNA expression of alkaline phosphatase (ALP), osteocalcin (OCN), and osteopontin (OPN). In addition, cell bio‐mineralization was evaluated through alizarin red S staining, and release of tumor necrosis factor alpha (TNF‐α), interleukin‐1 beta (IL‐1β), and interleukin‐6 (IL‐6) was measured through enzyme‐linked immunosorbent assays (ELISA). Furthermore, mRNA expression of Smad1, Smad4, and Smad5 and protein expression of phosphorylated Smad1, Smad4, and Smad5 were measured by reverse transcriptase polymerase chain reaction (RT‐PCR) and western blotting.ResultsThe ceramic debris had irregular shapes and sizes, and analysis of the size distribution using a particle size analyzer indicated that approximately 90% of the ceramic debris was smaller than 3.2 μm (2.0 ± 0.4 μm), which is considered clinically relevant. The results for mRNA expression of ALP, OCN, and OPN and alizarin red S staining indicated that cell differentiation and bio‐mineralization were significantly inhibited by the presence of ceramic debris at all tested concentrations (P < 0.05, and the values decreased gradually with the increase of ceramic debris concentration), but the results of the CCK‐8 assay showed that cell proliferation was not significantly affected (P > 0.05; there was no significant difference between the groups at 1, 3, and 5 days). In addition, the results of ELISA, RT‐PCR, and western blotting demonstrated that ceramic debris significantly promoted the release of inflammatory factors, including TNF‐α, IL‐β, and IL‐6 (P < 0.05, and the values increased gradually with the increase of ceramic debris concentration), and also greatly reduced the mRNA expression of Smad1, Smad4, and Smad5 (the values decreased gradually with the increase of ceramic debris concentration) as well as protein expression of phosphorylated Smad1, Smad4, and Smad5.ConclusionCeramic debris may affect differentiation and bio‐mineralization of MC3T3‐E subclone 14 cells through the bone morphogenetic protein/Smad signaling pathway.

Highlights

Total hip arthroplasty (THA) is an effective surgical procedure for treating hip disorders such as osteoarthritis, rheumatoid arthritis, and avascular necrosis, dramatically improving the quality of life of patients as a result of immediate relief from pain in the hip joint
The results for mRNA expression of alkaline phosphatase (ALP), OCN, and OPN and alizarin red S staining indicated that cell differentiation and bio-mineralization were significantly inhibited by the presence of ceramic debris at all tested concentrations (P < 0.05, and the values decreased gradually with the increase of ceramic debris concentration), but the results of the CCK-8 assay showed that cell proliferation was not significantly affected (P > 0.05; there was no significant difference between the groups at 1, 3, and 5 days)
The results of enzyme-linked immunosorbent assays (ELISA), reverse transcriptase polymerase chain reaction (RT-PCR), and western blotting demonstrated that ceramic debris significantly promoted the release of inflammatory factors, including TNF-α, IL-β, and IL-6 (P < 0.05, and the values increased gradually with the increase of ceramic debris concentration), and greatly reduced the mRNA expression of Smad[1], Smad[4], and Smad[5] as well as protein expression of phosphorylated Smad[1], Smad[4], and Smad[5]

Summary

Introduction

Total hip arthroplasty (THA) is an effective surgical procedure for treating hip disorders such as osteoarthritis, rheumatoid arthritis, and avascular necrosis, dramatically improving the quality of life of patients as a result of immediate relief from pain in the hip joint. The first tentative methods of restoration of function for patients with an arthritic hip were developed by researchers through many novel ideas, but a significant contribution to the routine success achieved today is obviously the concept of low-friction arthroplasty proposed by John Charnley[1]. There are two types of methods currently available to fix the femoral stem; namely, the cemented method and the cementless method. Both the head–cup interface and the stem–cement interface are potential sites for the generation of wear debris[4–6]. Debonding of the stem–cement interface is considered inevitable for almost all stem designs, and a low-amplitude oscillatory micromotion will occur at this interface, resulting in fretting wear and the corresponding formation of wear debris. The increasing significance of the stem–cement interface to a certain degree may be due to the great wear reduction achieved by the use of metal-on-metal and ceramic-on-ceramic articulations for the head–cup interface[9,10]

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Conclusion