Elsevier

Biomaterials

Volume 27, Issue 15, May 2006, Pages 3010-3020
Biomaterials

Over-expression of p53/BAK in aseptic loosening after total hip replacement

https://doi.org/10.1016/j.biomaterials.2006.01.006Get rights and content

Abstract

Particle-induced osteolysis is a major cause of aseptic loosening after total joint replacement. The possible induction of apoptosis has not been addressed in great detail. Thus far, it has been shown that ceramic and polyethylene particles can induce apoptosis of macrophages in vitro. The purpose of this study was to test the hypothesis that wears debris generated from total hip arthroplasty could induce cellular damage and apoptosis in vivo. We therefore determined by immunohistochemical methods if increased expression of p53, an important transcription factor, and BAK and Bcl-2, two important regulators of apoptosis, can be found in interface membranes and capsules of hips with aseptically loose implants.

Strongly positive immunohistochemical staining for p53 and BAK was found in peri-implant tissues from patients with aseptic hip implant loosening. Differentiation of various cell types showed that macrophages stained positive for p53 in all capsule and interface specimens. p53 was frequently detected in giant cells. Positive staining of BAK in macrophages and giant cells was seen in all specimens. Some positive reactions were observed in fibroblasts, only two of 19 cases stained for p53 and three cases for BAK within synovial cells. Positive macrophages and giant cells were localized around polyethylene particles. While T-lymphocytes showed a regular BAK-staining, the other leukocytes were negative. Statistical analyses showed significant positive correlations (p<0.001) between the presence of polyethylene and metal debris and the expression of BAK and p53. Polyethylene particles were surrounded by more positive macrophages and giant cells than were metal particles, indicating that polyethylene debris may be a stronger inductor of cell cycle arrest and apoptosis than metal debris.

In this study apoptosis of macrophages, giant cells and T-lymphocytes in capsules and interface membranes of patients with aseptic hip implant loosening has been demonstrated in vivo. It is possible that the apoptotic cascade could evolve as a novel therapeutic target to prevent particle-induced osteolysis.

Introduction

Particle-induced osteolysis is a major cause of aseptic loosening after total joint replacement. Periprosthetic osteolysis is initiated by an aseptic inflammatory response to phagocytosis of implant wear particles resulting in increased proliferation and differentiation of osteoclast precursors into mature osteoclasts [1]. Progressive osteolysis can result in implant instability and failure, eventually requiring revision surgery. The complexity of this topic is reflected by the large number of published studies on particle-induced bio-reactivity. While the osteolytic cascade initiated by cytokine release from macrophages has been studied extensively, the possible induction of apoptosis has not been addressed in great detail [2], [3]. Thus far, it has been shown only in vitro that ceramic, metal and polyethylene particles can induce apoptosis of macrophages [4], [5].

p53 functions as a transcription factor, i.e. as a modulator which can turn crucial genes either on or off. It also inhibits DNA replication and is a checkpoint control molecule of the cell cycle. Furthermore the p53 tumour suppressor gene is an important regulator of apoptosis [6]. Previous studies have demonstrated expression of p53 in rheumatoid arthritis synovium [7], [8].

BAK is a member of the Bcl-2 family of homologous proteins which play a role in regulating apoptosis. The Bcl-2 related proteins interact with one another through the formation of homo- and heterodimers. The susceptibility of cells to apoptotic stimuli is thought to be controlled by the relative ratios of the different Bcl-2 family proteins [9], [10]. BAK has been demonstrated to accelerate the rate of apoptosis in growth factor deprived murine lymphoid, neuronal and fibroblastic cell lines [11], [12], [13]. The BAK protein binds Bcl-XL and Bcl-2, and is thought to induce apoptosis by counteracting the protective functions of Bcl-2 and Bcl-XL. Altered levels of BAK expression have been reported in various malignancies and inflammatory conditions.

Bcl-2 oncoprotein is a blocker of apoptotic cell death. Gene transfer experiments have shown that elevated levels of this protein can protect a wide variety of cells from diverse cell death stimuli ranging from growth factor withdrawal and cytotoxic lymphokines to virus infection and DNA damage, anticancer drugs and radiation [14], [15]. Bcl-2 oncoprotein resides on the cytoplasmic side of the mitochondrial outer membrane, endoplasmic reticulum and nuclear envelope [14], [16].

The purpose of this study was to determine if wear debris in interface membranes and capsules with aseptically loose total hip implants has the capacity to damage cells and induce cell cycle arrest and apoptosis, as documented by an increased presence of p53 and BAK and decreased presence of Bcl-2.

Section snippets

Patients

The study was approved by the Institutional Review Board of the University. We distinguished three groups. Group 1 consisted of total hip arthoplasty cases while Groups 2 and 3 served as controls.

Group 1: For clinical data of Group 1 see Table 1. The reason for revision surgery was aseptic loosening of a hip implant. The presence of infection was excluded by analysis of inflammatory markers in the blood (c-reactive protein, leucocytosis), and by culture of joint fluid obtained by aspiration

Histology

Group 1: HE staining of the interface tissue showed typical features of membranes that form in response to arthroplasty, i.e. numerous fibrous cells in a bed of connective tissue. In the capsule tissue HE staining showed the fibrous membrane with collagen fibres, and the synovial membrane with loose connective tissue, fibrous and synovial cells.

Varying quantities of wear debris, including metal and polyethylene particles, were found in the capsule and interface tissues, accompanied by a

Hypothesis

The purpose of this study was to evaluate if wear debris generated from total hip arthroplasty can induce cellular damage and apoptosis in vivo as documented by the presence of p53 and BAK in the tissue. According to a previous report, detection of p53 in the interface membranes from aseptically loose total hip replacements was not possible by means of Western blotting [20]. We preferred to use immunohistochemical techniques and detected a strong expression of p53 protein in the tissues of

Conclusions

In this study apoptosis of macrophages, giant cells and T-lymphocytes in capsules and interface membranes of patients with aseptic hip implant loosening has been detected in vivo. It is possible that the apoptotic cascade could become a new therapeutic target to prevent particle-induced osteolysis.

Acknowledgements

This study was supported by a grant from the Verein der Freunde und Förderer des Evangelischen Krankenhauses Essen-Werden e.V.

References (39)

  • D. Granchi et al.

    Cell death induced by metal ions: necrosis or apoptosis?

    J Mater Sci: Mater Med

    (1998)
  • C. Nieder et al.

    The challenge of p53 as prognostic and predictive factor in Hodgkin's or non-Hodgkin's lymphoma

    Ann Hematol

    (2001)
  • G.S. Firestein et al.

    Apoptosis in rheumatoid arthritis: p53 overexpression in rheumatoid arthritis synovium

    Am J Pathol

    (1996)
  • P.P. Tak et al.

    p53 overexpression in synovial tissue from patients with early and longstanding rheumatoid arthritis compared with patients with reactive arthritis and osteoarthritis

    Arthritis Rheum

    (1999)
  • J.C. Reed

    Mini-review: cellular mechanisms of disease series. Bcl-2 and the regulation of programmed cell death

    J Cell Biol

    (1994)
  • T. Sato et al.

    Interactions among members of the Bcl-2 protein family analyzed with a yeast two-hybrid system

    PNAS USA

    (1994)
  • T. Chittenden et al.

    Induction of apoptosis by the Bcl-2 homologue Bak

    Nature

    (1995)
  • S.N. Farrow et al.

    Cloning of a bcl-2 homologue by interaction with adenovirus E1B 19 K

    Nature

    (1995)
  • M.C. Kiefer et al.

    Modulation of apoptosis by the widely distributed Bcl-2 homologue Bak

    Nature

    (1995)
  • Cited by (26)

    • Contributions of human tissue analysis to understanding the mechanisms of loosening and osteolysis in total hip replacement

      2014, Acta Biomaterialia
      Citation Excerpt :

      The immunostains used were based on the reaction between antigen and primary and secondary antibodies, with one of them being labeled with an enzyme (horseradish peroxidase, alkaline phosphatase, biotin), the fluorophore fluorescein isothiocyanate [33,34] or tetramethylrhodamine isothiocyanate [34]. Immunoenzyme protocols with many different principles were applied for antibody-aided detection, including: (i) the avidin–biotin complex method [19,25,35–51]; (ii) the labeled streptavidin–biotin method [32,33,52–59]; and (iii) the polymer-based detection method [60–67]. The presence of antigen was most often visualized by chromogen 3,3′-diaminobenzidine tetrachloride, which produces a brown reaction that can be seen with a light microscope.

    • Adiponectin attenuates osteolysis in aseptic loosening of total hip replacements

      2014, Acta Biomaterialia
      Citation Excerpt :

      However, knowledge about the role of apoptosis in this context is limited. Previous studies have demonstrated that apoptosis in macrophages and giant cells during particle engulfment is partially responsible for osteolysis in aseptic loosening of joint implants [2–4]. We demonstrated by means of a murine calvarial model of wear particle-induced osteolysis that inhibition of apoptosis leads to decreasing bone resorption by osteoclasts [5].

    • Arthroscopic surgical tools: A source of metal particles and possible joint damage

      2013, Arthroscopy - Journal of Arthroscopic and Related Surgery
    • Senescence in cells in aseptic loosening after total hip replacement

      2011, Acta Biomaterialia
      Citation Excerpt :

      Further studies are necessary to clarify if senescence in the cells of patients with aseptic loosening is induced by telomeric DNA damage as the consequence of an increased replication rate or by non-telomeric DNA damage or by both. In previous studies we have already reported that giant cells and macrophages can also react by transient cell cycle arrest and apoptosis to particle-induced cell damage [27,28]. The apoptotic reactions again result in increased bone resorption [29].

    View all citing articles on Scopus

    Investigation performed at the University of Duisburg-Essen, Germany.

    View full text