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Background Recent studies indicate that, in addition to antibody production, lymphocyte responses to SARS-CoV-2 may play an important role in protective immunity to COVID-19 and a percentage of the general population may exhibit lymphocyte memory due to unknown/asymptomatic exposure to SARS-CoV-2 or cross-reactivity to other more common coronaviruses pre-vaccination. Total joint replacement (TJR) candidates returning to elective surgeries (median age 68 years) may exhibit similar lymphocyte and/or antibody protection to COVID-19 prior to vaccination Methods In this retrospective study, we analyzed antibody titters, lymphocyte memory, and inflammatory biomarkers specific for the Spike and Nucleocapsid proteins of the SARS-CoV-2 virus in a cohort of n=73 returning TJR candidates (knees and/or hips) pre-operatively. Results Peripheral blood serum of TJR candidate patients exhibited a positivity rate of 18.4% and 4% for IgG antibodies specific for SARS-CoV-2 nucleocapsid and spike proteins, respectively. 13.5% of TJR candidates exhibited positive lymphocyte reactivity (SI > 2) to the SARS-CoV-2 nucleocapsid protein and 38% to the spike protein. SARS-CoV-2 reactive lymphocytes exhibited a higher production of inflammatory biomarkers (i.e., IL-1β, IL-6, TNFα, and IL-1RA) compared to non-reactive lymphocytes. Conclusions A percentage of TJR candidates returning for elective surgeries exhibit pre-vaccination positive SARS-CoV-2 antibodies and T cell memory responses with associated pro-inflammatory biomarkers. This is an important parameter for understanding immunity, risk profiles, and may aid pre-operative planning. Trial registration Retrospectively registered.

Background A battlefield-related injury results in increased local and systemic innate immune inflammatory responses, resulting in wound-specific complications and an increased incidence of osteoarthritis. However, little is known about whether severe injuries affect long-term systemic homeostasis, for example, immune function. Moreover, it also remains unknown whether battlefield-acquired metal fragments retained over the long term result in residual systemic effects such as altered immune reactivity to metals. Questions/purposes Does a retained metal fragment from a battlefield injury contribute to increased (1) adaptive metal-specific immune responses, (2) systemically elevated metal ion serum levels, and (3) serum immunoglobulin levels compared with combat injuries that did not result in a retained metal fragment? Methods In this pilot study, we analyzed metal-immunogenicity in injured military personnel and noninjured control participants using lymphocyte transformation testing (LTT, lymphocyte proliferation responses to cobalt, chromium and nickel challenge at 0.001, 0.01 and 0.1-mM concentrations in triplicate for each participant), serum metal ion analysis (ICP-mass spectroscopy), and serum immunoglobulin analysis (IgE, IgG, IgA, and IgM ). Military personnel with a battlefield-sustained injury self-recruited without any exclusion for sex, age, degree of injury. Those with battlefield injury resulting in retained metal fragments (INJ-FRAG, n = 20 male, mean time since injury ± SD was 12 ± 10 years) were compared with those with a battlefield injury but without retained metal fragments (INJ-NO-FRAG, n = 12 male, mean time since injury ± SD was 13 ± 12 years). A control group comprised of male noninjured participants was used to compare measured immunogenicity metrics (n = 11, males were selected to match battlefield injury group demographics). Results Military participants with sustained metal fragments had increased levels of metal-induced lymphocyte responses. The lymphocyte stimulation index among military participants with metal fragments was higher than in those with nonretained metal fragments (stimulation index = 4.2 ± 6.0 versus stimulation index = 2.1 ± 1.2 (mean difference 2.1 ± 1.4 [95% confidence interval 5.1 to 0.8]; p = 0.07) and an average stimulation index = 2 ± 1 in noninjured controls. Four of 20 participants injured with retained fragments had a lymphocyte proliferation index greater than 2 to cobalt compared with 0 in the group without a retained metal fragment or 0 in the control participants. However, with the numbers available, military personnel with retained metal fragments did not have higher serum metal ion levels than military participants without retained metal fragment-related injuries or control participants. Military personnel with retained metal fragments had lower serum immunoglobulin levels (IgG, IgA, and IgM) than military personnel without retained metal fragments and noninjured controls, except for IgE. Individuals who were metal-reactive positive (that is, a stimulation index > 2) with retained metal fragments had higher median IgE serum levels than participants who metal-reactive with nonmetal injuries (1198 ± 383 IU/mL versus 171 ± 67 IU/mL, mean difference 1027 ± 477 IU/mL [95% CI 2029 to 25]; p = 0.02). Conclusions We found that males with retained metal fragments after a battlefield-related injury had altered adaptive immune responses compared with battlefield-injured military personnel without indwelling metal fragments. Military participants with a retained metal fragment had an increased proportion of group members and increased average lymphocyte reactivity to common implant metals such as nickel and cobalt. Further studies are needed to determine a causal association between exposure to amounts of retained metal fragments, type of injury, personnel demographics and general immune function/reactivity that may affect personal health or future metal implant performance. Level of evidence Level IV, therapeutic study.

It is widely recognized that innate macrophage immune reactions to implant debris are central to the inflammatory responses that drive biologic implant failure over the long term. Less common, adaptive lymphocyte immune reactions to implant debris, such as delayed type hypersensitivity (DTH), can also affect implant performance. It is unknown which key patient factors, if any, mediate these adaptive immune responses that potentiate particle/macrophage mediated osteolysis. The objective of this investigation was to determine to what degree known adaptive immune responses to metal implant debris can affect particle-induced osteolysis (PIO); and if this pathomechanism is dependent on: 1) innate immune danger signaling, i.e., NLRP3 inflammasome activity, 2) sex, and/or 3) age. We used an established murine calvaria model of PIO using male and female wild-type C57BL/6 vs. Caspase-1 deficient mice as well as young (12–16 weeks old) vs. aged (18–24 months old) female and male C57BL/6 mice. After induction of metal-DTH, and Cobalt-alloy particle (ASTM F-75, 0.4um median diameter) calvaria challenge, bone resorption was assessed using quantitative micro-computed tomography (micro-CT) analysis and immune responses were assessed by measuring paw inflammation, lymphocyte transformation test (LTT) reactivity and adaptive immune cytokines IFN-gamma and IL-17 (ELISA). Younger aged C57BL/6 female mice exhibited the highest rate and severity of metal sensitivity lymphocyte responses that also translated into higher PIO compared to any other experimental group. The absence of inflammasome/caspase-1 activity significantly suppressed DTH metal-reactivity and osteolysis in both male and female Caspase-1 deficient mice. These murine model results indicate that young female mice are more predisposed to metal-DTH augmented inflammatory responses to wear debris, which is highly influenced by active NLRP3 inflammasome/caspase-1 danger signaling. If these results are clinically meaningful for orthopedic patients, then younger female individuals should be appropriately assessed and followed for DTH derived peri-implant complications.

Background: The rate of revision for some designs of total hip replacements due to idiopathic aseptic loosening has been reported as higher for women. However, whether this is environmental or inherently sex-related is not clear. Objective: Can particle induced osteolysis be sex dependent? And if so, is this dependent on the type of implant debris (e.g. metal vs polymer)? The objective of this study was to test for material dependent inflammatory osteolysis that may be linked to sex using CoCrMo and implant grade conventional polyethylene (UHMWPE), using an in vivo murine calvaria model. Methods: Healthy 12 week old female and male C57BL/6J mice were treated with UHMWPE (1.0um ECD) or CoCrMo particles (0.9um ECD) or received sham surgery. Bone resorption was assessed by micro-computed tomography, histology and histomorphometry on day 12 post challenge. Results: Female mice that received CoCrMo particles showed significantly more inflammatory osteolysis and bone destruction compared to the females who received UHMWPE implant debris. Moreover, females challenged with CoCrMo particles exhibited 120% more inflammatory bone loss compared to males (p Conclusion: We demonstrated sex-specific differences in the amount of osteolysis resulting from CoCrMo particle challenge. This suggests osteo-immune responses to metal debris are preferentially higher in female compared to male mice, and supports the contention that there may be inherent sex related susceptibility to some types of implant debris.

Options to treat early-onset scoliosis include guided-growth systems with sliding action between rods and pedicle screws. The wear was previously measured in an in vitro test, and in this in vivo rabbit model, we evaluated the local and systemic biological response to the stainless steel debris. Compared to the previous study, a relatively higher volume of representative wear particles with a median particle size of 0.84 μm were generated. Bolus dosages were injected into the epidural space at L4-L5 for a minimum of 36 rabbits across three treatment groups (negative control, 1.5 mg, and 4.0 mg) and two timepoints (12 and 24 weeks). Gross pathology evaluated distant organs and the injection site with a dorsal laminectomy to examine the epidural space and dosing site. Peri-implanted particle tissues were stained for immunohistochemical and quantitatively analyzed for IL-6 and TNF-α cytokines. Based on ISO 10993-6:2007 scoring, particles in the high-dose group were primarily non-irritant (12 weeks) with one slightly irritant. At 24 weeks, inflammatory cell infiltration was non-existent to minimal with all groups considered non-irritant at the injection site. Material characterization confirmed that particles detected in distant organs were stainless steel or contaminants. At 12 weeks, stainless steel groups demonstrated statistically increased amounts of cytokine levels compared to control but there was a statistical decrease for both at 24 weeks. These findings indicate that stainless steel wear debris, comparable to the expected usage from a simulated growth guidance system, had no discernible untoward biological effects locally and systemically in an animal model. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1980-1990, 2018.

Background The fretting-corrosion behavior of mixed metal contacts is affected by various mechanical and electrochemical parameters. Crevice conditions at the junction and patient-specific pathologies can affect the pH of the prosthetic environment. The main objective of this study is to understand the effect of pH variation at the stem/head junction of the hip implant under fretting corrosion exposure. We hypothesized that pH will have a significant influence on the fretting-corrosion behavior hip implant modular junctions. Materials and methods A custom-made setup was used to evaluate the fretting corrosion behavior of hip implant modular junctions. A Newborn calf serum solution (30 g/L protein content) was used to simulate the synovial fluid environment. A sinusoidal fretting motion, with a displacement amplitude of +50 µm, was applied to the Ti alloy rod. The effects of pathology driven, periprosthetic pH variation were simulated at four different pH levels (3.0, 4.5, 6.0 and 7.6). Electrochemical and mechanical properties were evaluated before, during, and after the applied fretting motion. Results The impedance of the system was increased in response to the fretting motion. The hysteresis tangential load/displacement behavior was not affected by pH level. The worn surfaces of CoCrMo pins exhibited the presence of tribolayer or organic deposits, in the pH 4.5 group, which may explain the lower drop in potential and mass loss observed in that group. Mechanically dominated wear mechanisms, namely, adhesive wear was shown in the pH 7.6 group, which may account for a higher potential drop and metal content loss. Conclusions This study suggests that the fretting-corrosion mechanisms in hip implant are affected by the pH levels of the surrounding environment and patient-specific factors.

Posterior spinal fusion implants include number of interconnecting components, which are subjected to micromotion under physiological loading conditions inducing a potential for fretting corrosion. There is very little known about the fretting corrosion in these devices in terms of the minimum angular displacement (threshold) necessary to induce fretting corrosion or the amount of fretting corrosion that can arise during the life of the implant. Therefore, the first goal was to evaluate the threshold fretting corrosion in three anatomical orientations and second the long-term fretting corrosion for the three different material types of spinal implants under physiological loading conditions. In threshold test, axial rotation exhibited highest changes in open circuit potential (VOCP in mV) and induced fretting currents (Ifrett in µA) for cobalt chrome (ΔVOCP : 24.71 ± 5.53; ΔIfrett : 4.03 ± 0.51) and stainless steel (ΔVOCP : 28.21 ± 6.97; ΔIfrett : 2.98 ± 0.42) constructs whereas it was flexion-extension for titanium constructs (ΔVOCP : 4.51 ± 2.48; ΔIfrett : 0.38 ± 0.12). Long-term test indicated that the titanium (VOCP :101 ± 0.06; Ifrett : 0.07 ± 0.02) and cobalt chrome (VOCP : 140.67 ± 0.04; Ifrett : 0.12 ± 0.05) constructs were more resistant to the fretting corrosion compared to stainless steel (VOCP : -135.33 ± 0.31; Ifrett : 2.63 ± 1.06). © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2858-2868, 2018.

Contemporary hip implants feature a modular design. Increased reported failure rates associated with the utilization of modular junctions have raised many clinical concerns. Typically, these modular interfaces contain circumferential machining marks (threads or microgrooves), but the effect of the machining marks on the fretting-corrosion behavior of total hip implant materials is unknown. This study reports the effects of microgrooves on the fretting-corrosion behavior of hip implant materials. The flat portions of two cylindrical, polished, CrCrMo alloy pins were loaded horizontally against one rectangular Ti alloy rod. Two surface preparation groups were used for the Ti6Al4V rod (polished and machined). The polished group was prepared using the same methods as the CoCrMo pins. The machined samples were prepared by creating parallel lines on the rod surfaces to represent microgrooves present on the stem tapers of head-neck modular junctions. Newborn calf serum (30 g/L protein content; 37 °C) at pH of levels of 7.6 and 3.0 were used to simulate the normal joint fluid and a lowered pH within a crevice, respectively. The samples were tested in a fretting corrosion apparatus under a 200N normal force and a 1Hz sinusoidal fretting motion with a displacement amplitude of 25 μm. All electrochemical measurements were performed with a potentiostat in a three-electrode configuration. The results show significant differences between machined samples and polished samples in both electrochemical and mechanical responses. In all cases, the magnitude of the drop in potential was greater in the machined group compared to the polished group. The machined group showed a lower total dissipated friction energy for the entire test compared to the polished group. Additionally, the potentiostatic test measurements revealed a higher evolved charge in the machined group compared to the polished group at both pH conditions (pH 7.6 and 3.0). The machined surfaces lowered the overall dissipated friction energy at pH 7.6 compared to pH 3.0, but also compromised electrochemical performance in the tested conditions. Therefore, the role of synergistic interaction of wear and corrosion with surface topographical changes is evident from the outcome of the study. Despite the shift towards higher electrochemical destabilization in the machined group, both polished and machined groups still exhibited a mechanically dominated degradation.

The pathology of early aseptic implant failure has been linked in basic science, case, and group studies to metal implant debris–induced delayed-type hypersensitivity (DTH) responses (or metal allergy). However, a precondition of metal DTH does not always result in implant failure. The methods of diagnosing metal DTH responses measure cell-mediated adaptive immunity, where conditioned lymphocytes respond to specific stimuli. Metal-sensitivity testing is a direct test of an individual’s immune response to metal challenge, which has been in use for over the past half century to measure delayed-type responses to drugs (such as antibiotics) where the results indicate levels of immune reactivity. Currently approved methods for human diagnostic testing for metal DTH include both skin testing (patch testing) and in vitro blood testing using lymphocyte transformation testing (LTT). LTT testing may be better suited for diagnosing implant-related sensitivity, given the ability of dermal exposure to metals to induce DTH responses. Metal sensitivity among the general population has been well established with a rate of 10%-15%, where hypersensitivity to nickel is the most common. However, it remains unknown about the exact percentage of these presensitized people that will go on to develop pathogenic metal-sensitivity responses to their implant. Recent studies show that when exposure to metal implant debris is minimized, the chances of metal DTH reactivity are also minimized. Further, past basic science studies, case studies, and cohort studies indicate the clinical utility of metal-sensitivity testing in specific patient populations. Continuing clinical outcome studies are needed to relate positive patch and metal-LTT test results with other clinical determinants to more definitively predict and prevent pathologic reactivity to implants.

BACKGROUND Recent studies indicate that females demonstrate an increased risk of experiencing adverse local tissue reactions, aseptic loosening, and revision after primary metal-on-metal hip resurfacing arthroplasty compared with males; the underlying biological mechanisms responsible for sex discrepancies in implant failure remain unclear. In addition to anatomical and biomechanical sex differences, there may be inherent immunological disparities that predispose females to more aggressive adaptive immune reactivity to implant debris, i.e., metal sensitivity. METHODS In this retrospective study, we analyzed sex-associated rates and levels of metal sensitization in 1,038 male and 1,575 female subjects with idiopathic joint pain following total joint arthroplasty (TJA) who were referred for in vitro metal-sensitivity testing. RESULTS Females demonstrated a significantly higher rate and severity of metal sensitization compared with males. The median lymphocyte stimulation index (SI) among males was 2.8 (mean, 5.4; 95% confidence interval [CI], 4.9 to 6.0) compared with 3.5 (mean, 8.2; 95% CI, 7.4 to 9.0) among females (p < 0.05). Forty-nine percent of females had an SI of ≥4 (reactive) compared with 38% of males, and the implant-related level of pain was also significantly (p < 0.0001) higher among females (mean, 6.8; 95% CI, 6.6 to 6.9) compared with males (mean, 6.1; 95% CI, 6.0 to 6.3). CONCLUSIONS In a select group of patients who had joint pain following TJA and no evidence of infection and who were referred for metal-sensitivity testing, females exhibited a higher level of pain and demonstrated a higher rate and severity (as measured by lymphocyte SI) of metal sensitization compared with males. LEVEL OF EVIDENCE Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.

There has been an increased interest in the role of corrosion in total hip arthroplasty. This is based on reports of early implant failures and adverse local tissue reaction resulting from excessive corrosion at the modular interfaces of some implant designs. Orthopedic alloys are not selected based solely on their mechanical properties of strength, but rather because they possess the best balance between material properties of corrosion resistance and mechanical properties of strength. The passive layer of a metal oxide that develops on a surface of a metal serves a critical role in preventing corrosion. However, this protective layer is a dynamic structure. Aggressive corrosion occurs on implants when the kinetics of this oxide layer’s destruction dominates over its generation. There is a spectrum of different types of corrosion defined by the environment and stability of the passive layer. Pitting is the localized dissolution of this protective metal oxide film. Crevice corrosion occurs with a similar mechanism but in an isolated environment that promotes corrosion. Fretting corrosion or mechanically assisted crevice corrosion (MACC) occurs with the addition of mechanical oscillating loads at the modular junction that disrupts this protective layer. Understanding this process is important to improve implant designs, surgical technique, and assessment of patients.

Wear and corrosion in total hip replacement negatively impact implant service-life and patient well-being. The aim of this study was to generate a statistical response surface of material loss using an apparatus, capable of testing the effect of wear and corrosion products in situ on cells, such as macrophages. The test chamber of a ball-on-flat tribometer operating inside a CO2 incubator was integrated with an electrochemical setup and adapted for cell culture work. A 20-test series, following a 2-level 3-factor design of experiments, was performed with a ceramic head in reciprocating rotational motion against a CoCrMo-alloy disc, under constant load. The lubricant was cell culture medium (RPMI-1640 + 10 vol% bovine serum). Response surfaces were generated, which statistically showed the influence of motion amplitude, load, and potential on the total mass loss and wear scar volume of the metallic discs. Potential had the highest impact on the total mass loss, while motion amplitude and load significantly influenced the wear scar volume. The concentrations of the alloy elements found in the lubricants reflected the bulk-alloy stoichiometry. The total concentration of Co released into the lubricant (2.3–63 ppm by total mass loss, 1.5 to 62 ppm by ICP-MS) corresponded well with the known range to trigger cell response. Tribocorrosion tests in the presence of cells and tissues, such as macrophages, lymphocytes and/or synovium, will be carried out in the future.

Currently, there is a dearth of information regarding the degree of particle shedding from breast implants (BIs) and what are the general biological consequences of BI debris. Thus, it is unclear to what degree BI debris compromises the long-term biological performance of BIs. For orthopedic implants, it is well established that the severity of biological reactivity to implant debris governs long-term clinical performance. Orthopedic implant particulate debris is generally in the range of 0.01 to 100 μm in diameter. Implant debris-induced bioreactivity/inflammation is mostly a peri-implant phenomenon caused by local innate immune cells (eg, macrophages) that produce proinflammatory cytokines such as tumor necrosis factor-α, interleukin-1β, interleukin-6, and prostaglandin 2 (PGE2). In orthopedics, there have been few systemic concerns associated with polymeric implant debris (like silicone) other than documented dissemination to remote organs (eg, liver, spleen, etc.) with no known associated pathogenicity. This is not true of metal implant debris where normal (well-functioning) implants can induce systemic reactions such as delayed type hypersensitivity. Diagnostic analysis of orthopedic tissues has focused on innate (macrophage mediated) and adaptive (lymphocyte-mediated hypersensitivity) immune responses. Orthopedic implant debris-associated lymphocyte cancers have not been reported in over 40 years of orthopedic literature. Adaptive immune responses such as hypersensitivity reactions to orthopedic implant debris have been dominated by certain implant types that produce specific kinds of debris (eg, metal-on-metal total joint prostheses). Orthopedic hypersensitivity responses and atypical BI bioreactivity such as BI-associated anaplastic large cell lymphoma share crossover markers for diagnosis. Differentiating normal innate immune reactivity to particles from anaplastic large cell lymphoma reactions from delayed type hypersensitivity reactions to BI-associated implant debris remains unclear but vital to patients and surgeons.

Background Recently, there has been increasing concern in the orthopedic community over the use of hip implant modular devices due to an increasing number of reports of early failure, failure that has been attributed to fretting-corrosion at modular interfaces. Much is still unknown about the electrochemical and mechanical degradation mechanisms associated with the use of such devices. Purpose Accordingly, the purpose of our study was to develop a methodology for testing the fretting-corrosion behavior of modular junctions. Methods A fretting-corrosion apparatus was used to simulate the fretting-corrosion conditions of a CoCrMo hip implant head on a Ti6Al4V hip implant stem. The device features two perpendicularly-loaded CoCrMo pins that articulated against a Ti6Al4V rod. A sinusoidal fretting motion was applied to the rod at various displacement amplitudes (25, 50, 100, 150 and 200 μm) at a constant load of 200 N. Bovine calf serum at two different pH levels (3.0 and 7.6) was used to simulate the fluid environment around the joint. Experiments were conducted in two modes of electrochemical control – free-potential and potentiostatic. Electrochemical impedance spectroscopy tests were done before and after the fretting motion to assess changes in corrosion kinetics. Results In free potential mode, differences were seen in change in potential as a function of displacement amplitude. In general, V Drop (the drop in potential at the onset of fretting), V Fretting, (the average potential during fretting), Δ V Fretting (the change in potential from the onset of fretting to its termination) and V Recovery (the change in potential from the termination of fretting until stabilization) appeared linear at both pH levels, but showed drastic deviation from linearity at 100 μm displacement amplitude. Subsequent EDS analysis revealed a large number of Ti deposits on the CoCrMo pin surfaces. Potentiostatic tests at both pH levels generally showed increasing current with increasing displacement amplitude. Electrochemical impedance spectroscopy measurements from free potential and potentiostatic tests indicated increased levels of resistance of the system after induction of the fretting motion. In free potential tests, the largest increase in impedance was found for the 100 μm group. Conclusions We conclude that the 100 µm group exhibits deviations from linearity for several parameters, and this was most likely due to adhesive wear between Ti6Al4V and CoCrMo surfaces. Overall, the degradation of the system was dominated by wear at all pH levels, and displacement amplitudes.

Innate immune reactions to orthopedic implant debris are the primary cause of total joint replacement (TJR) failure over the long term (15-20 years). The role of pathogen associated pattern recognition receptors (i.e., TLRs) in regulating immune reactivity to metal implant particles remains controversial. Do different TLRs (i.e., TLR2 vs. TLR4) activated by their respective ligands in concert with metal implant debris elicit equivalent innate immune responses? In this investigation, our in vitro and in vivo data indicate that Gram-negative PAMPs are more pro-inflammatory than Gram-positive PAMPs. In vitro results indicated TLR4 activation in concert with CoCrMo orthopedic implant debris (CoCrMo/LPS+) challenged primary macrophages resulted in significantly greater inflammatory responses than CoCrMo/PAM3CSK+ (TLR2). Similarly, in vivo results indicated CoCrMo/LPS+ TLR4 challenge induced a twofold increase in inflammation-induced bone resorption (osteolysis) than CoCrMo/PAM3CSK+ (p

Periprosthetic osteolysis remains a major limitation of long-term successful total hip replacements with ultra-high molecular weight polyethylene (UHMWPE) bearings. As intra and extracellular reactive oxygen species are know to contribute to wear debris-induced osteoclastic bone resorption and decreased osteoblastic bone formation, antioxidant doped UHMWPE has emerged as an approach to reduce the osteolytic potential of wear debris and maintain coupled bone remodeling. To test this hypothesis in vivo, we evaluated the effects of crosslinked UHMWPE wear debris particles (AltrX(™) ), versus similar wear particles made from COVERNOX(™) containing UHMWPE (AOX(™) ), in an established murine calvaria model. Eight-week-old female C57B/6 mice (n = 10/Group) received a pre-op micro-CT scan prior to surgical implantation of the UHMWPE particles (2mg), or surgery without particles (sham). Dynamic labeling was performed by intraperitoneal injection of calcein on day 7 and alizarin on day 9, and the calvaria were harvested for micro-CT and histology on day 10. Surprisingly, we found that AOX particles induced significantly more bone resorption (1.72-fold) and osteoclast numbers (1.99-fold) vs. AltrX (p

Modern hip prostheses feature a modular implant design with at least one tapered junction. This design can lead to several complications due to the introduction of additional interfaces, which are subjected to various loading conditions and micromotion. The main objective of current study is to develop a fretting corrosion apparatus, which is able characterize the mechanical and electrochemical behaviour of various existing metal alloy couples during fretting motion. This study describes the design and the main considerations during the development of a novel fretting corrosion apparatus, as well as determination of the machine compliance and the initial testing results. Machine compliance considerations and frictional interactions of the couples are discussed in detail. For the preliminary tests, metal alloy pins, made of Ti6Al4V and wrought high-carbon CoCrMo were mechanically polished to a surface roughness of less than 20nm. 2 pins (Diameter = 11mm) of either Ti6Al4V or CoCrMo were loaded onto a Ti6Al4V alloy rod at a normal force of 200N. The interface types included: Ti6Al4V-Ti6Al4V-Ti6Al4V, Ti6Al4V-Ti6Al4V-CoCrMo, and CoCrMo-Ti6Al4V-CoCrMo. The Ti6Al4V rod articulated against the metal alloy pins in a sinusoidal fretting motion with a displacement amplitude of ±50μm. Bovine calf serum (30g/L of protein content) was selected as a lubricant and tested at 2 different pH levels (pH 3.0 and 7.6). In all cases, current and friction energy were monitored during the fretting process. The results indicated distinct, material-specific current evolutions and friction energies. No significant differences were observed in electrochemical or mechanical behaviour in response to pH change. In general, Ti6Al4V-Ti6Al4V-Ti6Al4V couples displayed the earliest passivation and superior electrochemical behaviour compared to Ti6Al4V-Ti6Al4V-CoCrMo and CoCrMo-Ti6Al4V-CoCrMo under fretting conditions. In addition, fluctuations in current were observed in specific regions at all instances where Ti6Al4V was coupled with Ti6Al4V. These fluctuations were not observed in instances where Ti6Al4V was coupled with CoCrMo. These findings suggest transitions in the degradation mechanisms at the modular junction as a function of material couples/contacts. The findings may assist in improving the current hip modular junctions.

Sterile solid microparticles (MPs) trigger an innate type 2 inflammatory response dependent on a SYK-BTK-IL33 signaling axis (Nature Materials,18, 289,2019). Inoculation with MPs trigger innate inflammation with influx of neutrophils, eosinophils and alternatively activated macrophages (M2). MPs also trigger cell death in macrophages leading to release of IL33 and the development of a type 2 response with increased IL4,IL5 and IL13 and expression of M2 markers including Arg1, Relmα and Chil3. Blockade of cell death by inhibiting necroptosis (necrox-2) or apoptosis (ZVED-FMK) alone or in combination inhibited the MP mediated release of IL33 by macrophages ex vivo. We also examined the role of adenosine in MP-mediated IL-33 release by inoculating LysMCreA2bfl/flwith MP.MP induced inflammation was blocked in LysMCreA2bfl/fmice compared to LysMCre controls, with significant decreases in type 2 cytokines and IL33. Adoptive transfer of wild type macrophages to LysMCreA2bfl/flrestored the MP-mediated type 2 inflammation. These findings indicate that programmed cell death and A2b receptor signaling pathways play a major role in particulate induced sterile inflammation.

Metal ion release from the orthopedic implants is a great concern for the clinicians and patients. Currently, the inductive coupled plasma-mass spectroscopy (ICP-MS) technique is used to detect and estimate the metal ions in the blood or synovial fluid, which is expensive and needs technical assistance. Hence, the aim of the current work is to develop a biosensor based on the electrochemistry to measure the metal release to the body fluids (blood or synovial fluid) from the implants. This will work very similar to a glucometer (by function), as a patient-driven technique, if it is optimized for the blood samples, and for clinical purpose, in case of the synovial fluid estimation. As a proof of concept effort, the present study has two objectives: (1) To study the effectiveness of using a micro-chip biosensor as a diagnostic technique for the early detection of the released metal particles in the synovial fluid solution (study 1), and (2) to investigate the corrosion kinetics of CoCrMo alloy in the presence of metal particles in synovial fluid solution (study 2). A series of tests were done with biosensor prototype with increasing concentration of metal release (particles and ions), which is generated from a hip simulator (study 1). The impedance variation (delta Z) shows a very close correlation with increased amount of metal release (particles and ions) level (study 2). Although the study has several limitations, the initial findings indicate that a biosensor could be developed as a diagnostic tool to detect the metal release (particles and ions) levels.

Initiation of the innate sterile inflammatory response that can develop in response to microparticle exposure is little understood. Here, we report that a potent type 2 immune response associated with the accumulation of neutrophils, eosinophils and alternatively activated (M2) macrophages was observed in response to sterile microparticles similar in size to wear debris associated with prosthetic implants. Although elevations in interleukin-33 (IL-33) and type 2 cytokines occurred independently of caspase-1 inflammasome signalling, the response was dependent on Bruton’s tyrosine kinase (BTK). IL-33 was produced by macrophages and BTK-dependent expression of IL-33 by macrophages was sufficient to initiate the type 2 response. Analysis of inflammation in patient periprosthetic tissue also revealed type 2 responses under aseptic conditions in patients undergoing revision surgery. These findings indicate that microparticle-induced sterile inflammation is initiated by macrophages activated to produce IL-33. They further suggest that both BTK and IL-33 may provide therapeutic targets for wear debris-induced periprosthetic inflammation. Microparticle wear debris from prosthetic implants following patient revision surgery is observed to induce a potent type 2 inflammatory response involving cytokine secretion by macrophages through a Bruton’s tyrosine kinase-dependent signalling pathway.

T he study by Catelas and colleagues adds to the current scientific consensus that adaptive immune responses to metals occurs in a subset of all people with metal implants. The study authors also found that the incidence of pseudotumors correlates with systemic evidence of metal-related delayed-type hypersensitivity (DTH) responses and elevated metal ion levels. The current study identifies alterations in circulating adaptive immune cells in people with elevated levels of metal debris and pseudotumors. These findings support past histological and T-cell function studies, indicating pseudotumors are more prevalent in people with DTH-type immune reactivity. This highlights the growing recognition of these phenomena as central to implant performance in some individuals/implant designs. Case and group studies [4, 5] indicate that once locally activated, adaptive immune inflammation to metal contributes to (if not, mediates) the pathogenesis of poor implant performance. Metal DTH reactivity is a lymphocyte mediated response, similar to a Type IV DTH reaction. Metal-on-metal (MoM) hip failures have been associated with classic DTH responses including periimplant activated lymphocyte accumulations (aseptic lymphocytic vasculitisassociated lesions, [ALVAL]), and dermal and functional lymphocyte hypersensitivity responses to metals [2, 7, 11]. Elevated metal exposure can also cause different forms of toxicity responses in tissue, bone, and immune cells, including cobalt-induced hypoxia-like responses, postulated as causing pseudotumors [10]. Lymphocyte reactivity (metal DTH responses/ALVAL) and pseudotumors can be caused separately [6, 8]. Currently, there are no accepted thresholds for systemic metal ion levels that necessarily indicate for pseudotumor formation or immune reactions [1, 7]; it remains an increased risk association. This CORR Insights is a commentary on the article ‘‘Do Patients With a Failed Metal-onmetal Hip Implant With a Pseudotumor Present Differences in Their Peripheral Blood Lymphocyte Subpopulations?’’ by Catelas and colleagues available at: DOI: 10.1007/ s11999-015-4466-8. The author certifies that she, or any members of her immediate family, has no funding or commercial associations (consultancies, stock ownership, equity interest, patent/licensing arrangements, etc.) that might pose a conflict of interest in connection with the submitted article. All ICMJE Conflict of Interest Forms for authors and Clinical Orthopaedics and Related Research editors and board members are on file with the publication and can be viewed on request. The opinions expressed are those of the writers and do not reflect the opinion or policy of CORR or The Association of Bone and Joint Surgeons. This CORR Insights comment refers to the article available at DOI: 10.1007/s11999015-4466-8. N. J. Hallab PhD (&) Department of Orthopedic Surgery, Rush University Medical Center, 1735 W Harrison, Cohn Research Bldg, #722, Chicago, IL 60612, USA e-mail: nhallab@rush.edu CORR Insights Published online: 13 October 2015 The Association of Bone and Joint Surgeons1 2015

Posterior spinal fusion implants include number of interconnecting components, which are subjected to micromotion under physiological loading conditions inducing a potential for fretting corrosion. There is very little known about the fretting corrosion in these devices in terms of the minimum angular displacement (threshold) necessary to induce fretting corrosion or the amount of fretting corrosion that can arise during the life of the implant. Therefore, the first goal was to evaluate the threshold fretting corrosion in three anatomical orientations and second the long-term fretting corrosion for the three different material types of spinal implants under physiological loading conditions. In threshold test, axial rotation exhibited highest changes in open circuit potential (V

Despite the success in returning people to health saving mobility and high quality of life, the over 1 million total joint replacements implanted in the US each year are expected to eventually fail after approximately 15–25 years of use, due to slow progressive subtle inflammation to implant debris compromising the bone implant interface. This local inflammatory pseudo disease state is primarily caused by implant debris interaction with innate immune cells, i.e., macrophages. This implant debris can also activate an adaptive immune reaction giving rise to the concept of implant-related metal sensitivity. However, a consensus of studies agree the dominant form of this response is due to innate reactivity by macrophages to implant debris danger signaling (danger-associated molecular pattern) eliciting cytokine-based and chemokine inflammatory responses. This review covers implant debris-induced release of the cytokines and chemokines due to activation of the innate (and the adaptive) immune system and how this leads to subsequent implant failure through loosening and osteolysis, i.e., what is known of central chemokines (e.g., IL-8, monocyte chemotactic protein-1, MIP-1, CCL9, CCL10, CCL17, and CCL22) associated with implant debris reactivity as related to the innate immune system activation/cytokine expression, e.g., danger signaling (e.g., IL-1β, IL-18, IL-33, etc.), toll-like receptor activation (e.g., IL-6, tumor necrosis factor α, etc.), bone catabolism (e.g., TRAP5b), and hypoxia responses (HIF-1α). More study is needed, however, to fully understand these interactions to effectively counter cytokine- and chemokine-based orthopedic implant-related inflammation.

All of the over 1 million total joint replacements implanted in the US each year are expected to eventually fail after 15–25 years of use, due to slow progressive subtle inflammation at the bone implant interface. This inflammatory disease state is caused by implant debris acting, primarily, on innate immune cells, that is, macrophages. This slow progressive pathological bone loss or “aseptic loosening” is a potentially life-threatening condition due to the serious complications in older people (>75 yrs) of total joint replacement revision surgery. In some people implant debris (particles and ions from metals) can influence the adaptive immune system as well, giving rise to the concept of metal sensitivity. However, a consensus of studies agrees that the dominant form of this response is due to innate reactivity by macrophages to implant debris where both danger (DAMP) and pathogen (PAMP) signalling elicit cytokine-based inflammatory responses. This paper discusses implant debris induced release of the cytokines and chemokines due to activation of the innate (and the adaptive) immune system and the subsequent formation of osteolysis. Different mechanisms of implant-debris reactivity related to the innate immune system are detailed, for example, danger signalling (e.g., IL-1β, IL-18, IL-33, etc.), toll-like receptor activation (e.g., IL-6, TNF-α, etc.), apoptosis (e.g., caspases 3–9), bone catabolism (e.g., TRAP5b), and hypoxia responses (Hif1-α). Cytokine-based clinical and basic science studies are in progress to provide diagnosis and therapeutic intervention strategies.

Metal sensitivity testing is generally the diagnosis method of last resort for aseptic painful implants with elevated inflammatory responses. However, the relationship between implant-related pain and implant-debris-related metal sensitization remains incompletely understood. Although a sensitivity to nickel alone has been used as a general measure of metal allergy, it may lack the specificity to correlate sensitivity to specific implant metals and thus to select a biologically appropriate implant material. In this retrospective study, we report the incidence of pain and nickel sensitivity in patients with total joint arthroplasties (TJAs) referred for metal sensitivity testing (n=2018). We also correlated the degree of nickel hypersensitivity to implant pain levels (none, mild, moderate, and high, using a scale of 0-10) and the incidence of sensitivity to alternative implant metals in highly nickel-reactive subjects. Most patients (79%) reported pain levels that were moderate to high regardless of implant age, whereas patients with severely painful TJAs had a statistically greater incidence of nickel sensitivity over the short-term post-operative period (≤4 years). Patients with moderate pain scores (4-7) and high pain scores (≥8) also exhibited significantly higher sensitivity to nickel compared to patients with no pain and no implant (controls) (p0.05). Highly nickel-sensitive subjects (SI8) also showed incidences of sensitization to alternative materials such as cobalt, chromium, or molybdenum (57%) or aluminum or vanadium alloy (52%). These data suggest that painful TJAs caused by metal sensitivity more likely occur relatively early in the post-operative period (≤4 years). The incidences of sensitivity to alternative implant metals in only a subset of nickel-reactive patients highlights the importance of testing for sensitization to all potential revision implant materials.

Metal hypersensitivity has been recognized as an adverse biologic reaction that can compromise total joint arthroplasty (TJA) performance. However, the etiology of metal hypersensitivity responses in TJAs remains unclear. Metal implant debris is known to act as a danger signal that drives NLRP3 inflammasome activation. It remains unknown if implant debris induced inflammasome activation regulates T cell lineage in TJA metal hypersensitivity responses. In this study, we show both in vivo and in vitro that the pathogenesis of metal hypersensitivity responses to implant debris are largely dependent on activation of the inflammasome/caspase-1 pathway and subsequent production of IL-17A/F by CD4+ T cells. Inhibiting either the inflammasome pathway or IL-17A bioactivity in vivo and in vitro (in vivo using NLRP3 and Caspase-1 deficient mice or in vitro using blocking agents such as Capase-1 inhibitor, IL-1Ra and anti-IL-17A), significantly (p

Long-term aseptic failures of joint replacements are generally attributed to implant debris-induced inflammation and osteolysis. This response is largely mediated by immune and bone cells (monocytes/macrophages and osteoclasts, respectively), that in the presence of implant debris (e.g. metal particles and ions), release pro-inflammatory cytokines such as IL-1β, TNF-α, and IL-6. The relative degree to which implant debris can illicit inflammatory response(s) from osteoclasts vs monocytes/macrophages is unknown, i.e. are osteoclasts a viable target for anti-inflammatory therapy for implant debris? We investigated relative monocyte versus osteoclast inflammatory responses in a side-by-side comparison using implant debris from the perspective of both danger signaling (IL-1β) and pathogenic recognition (TNF-α) reactivity (Challenge Agents: Cobalt-alloy, Titanium-alloy, and PMMA particles, 0.9-1.8um-dia ECD and Cobalt, and Nickel-ions 0.01-0.1mM, all with and without LPS priming). Human monocytes/macrophages reacted to implant debris with >100 fold greater production of cytokines compared to osteoclast-like cells. Particulate Co-alloy challenge induced >1000 pg/ml of IL-1β and TNF-α, in monocytes and 3000pg/mL IL-1β and TNF-α in monocytes/macrophages and

Object The introduction and utilization of motion-preserving implant systems for spinal reconstruction served as the impetus for this basic scientific investigation. The effect of unintended wear particulate debris resulting from micromotion at spinal implant interconnections and bearing surfaces remains a clinical concern. Using an in vivo rabbit model, the current study quantified the neural and systemic histopathological responses following epidural application of 11 different types of medical-grade particulate wear debris produced from spinal instrumentation. Methods A total of 120 New Zealand White rabbits were equally randomized into 12 groups based on implant treatment: 1) sham (control), 2) stainless steel, 3) titanium alloy, 4) cobalt chromium alloy, 5) ultra–high molecular weight polyethylene (UHMWPe), 6) ceramic, 7) polytetrafluoroethylene, 8) polycarbonate urethane, 9) silicone, 10) polyethylene terephthalate, 11) polyester, and 12) polyetheretherketone. The surgical procedure consisted of a midline posterior approach followed by resection of the L-6 spinous process and L5–6 ligamentum flavum, permitting interlaminar exposure of the dural sac. Four milligrams of the appropriate treatment material (Groups 2–12) was then implanted onto the dura in a dry, sterile format. All particles (average size range 0.1–50 μm in diameter) were verified to be endotoxin free prior to implantation. Five animals from each treatment group were sacrificed at 3 months and 5 were sacrificed at 6 months postoperatively. Postmortem analysis included epidural cultures and histopathological assessment of local and systemic tissue samples. Immunocytochemical analysis of the spinal cord and overlying epidural fibrosis quantified the extent of proinflammatory cytokines (tumor necrosis factor–α, tumor necrosis factor–β, interleukin [IL]–1α, IL-1β, and IL-6) and activated macrophages. Results Epidural cultures were negative for nearly all cases, and there was no evidence of particulate debris or significant histopathological changes in the systemic tissues. Gross histopathological examination demonstrated increased levels of epidural fibrosis in the experimental treatment groups compared with the control group. Histopathological evaluation of the epidural fibrous tissues showed evidence of a histiocytic reaction containing phagocytized inert particles and foci of local inflammatory reactions. At 3 months, immunohistochemical examination of the spinal cord and epidural tissues demonstrated upregulation of IL-6 in the groups in which metallic and UHMWPe debris were implanted (p < 0.05), while macrophage activity levels were greatest in the stainless-steel and UHMWPe groups (p < 0.05). By 6 months, the levels of activated cytokines and macrophages in nearly all experimental cases were downregulated and not significantly different from those of the operative controls (p > 0.05). The spinal cord had no evidence of lesions or neuropathology. However, multiple treatments in the metallic groups exhibited a mild, chronic macrophage response to particulate debris, which had diffused intrathecally. Conclusions Epidural application of spinal instrumentation particulate wear debris elicits a chronic histiocytic reaction localized primarily within the epidural fibrosis. Particles have the capacity to diffuse intrathecally, eliciting a transient upregulation in macrophage/cytokine activity response within the epidural fibrosis. Overall, based on the time periods evaluated, there was no evidence of an acute neural or systemic histopathological response to the materials included in the current project.

Recent investigations indicate that innate immune “danger-signaling” pathways mediate metal implant debris induced-inflammatory responses, e.g. NALP3 inflammasome. How the physical characteristics of particles, (size, shape and chemical composition) affect this inflammatory reactivity remains controversial. We examined the role of Cobalt-Chromium-Molybdenum (CoCrMo) alloy particle shape and size on human macrophage phagocytosis, lysosomal destabilization, and inflammasome activation. Round/smooth vs. irregularly shaped/rough CoCrMo-alloy particles of ~1µm and 6 to 7µm diameter were investigated for differential lysosomal damage and inflammasome activation in human monocytes/macrophages. While spherical/smooth 1µm CoCrMo-alloy particles did not measurably affect macrophage IL-1β production, irregular 1µm CoCrMo-alloy particles induced significant IL-1β increases over controls. Both round/smooth particles and irregular CoCrMo-alloy particles that were 6 to 7µ min size induced >10-fold increases in IL-1β production compared to similarly shaped smaller particles (p 3-fold increase in IL-1β vs. similarly sized round/smooth particles (at an equal dose, particles/cell). CoCrMo-alloy particle-size-induced IL-1β production was dependent on the lysosomal protease Cathepsin B, further supporting lysosomal destabilization as causative in inflammation. Phagocytosable larger/irregular shaped particles (6µm) demonstrated the greatest lysosomal destabilization (observed immunofluorescently) and inflammatory reactivity when compared on an equal dose basis (particles/cell) to smaller/spherical 1µm particles in vitro.

Almost 20% of joint replacement implants fail at 15 to 20 years. Reports suggest that systemic effects of metal-on-metal implants and local effects of total joint arthroplasty implants contributing to implant failure are immune system based. Sometimes implant wear debris can cause implant failure resulting from bone fracture, infection, or implant fracture/failure; most often, aseptic osteolysis or loosening leads to wear debris. Debris is produced by wear (primary) or by corrosion. Corrosion-chemical oxidation comprising reduction reactions involving electron transport-produces electrochemical degradation. Metallic implant degradation occurs when electrochemical dissolution and mechanical/physical wear are combined (i.e., tribocorrosion). With metal-on-metal implants, even with relatively low levels of wear and particle release, pathology caused by metal debris such as pseudotumor/fibrous tissue growth can lead to early implant failure.

Some tissues from metal-on-metal (MoM) hip arthroplasty revisions have shown evidence of adaptive-immune reactivity (i.e., excessive peri-implant lymphocyte infiltration/activation). We hypothesized that, prior to symptoms, some people with MoM hip arthroplasty will develop quantifiable metal-induced lymphocyte reactivity responses related to peripheral metal ion levels. We tested three cohorts (Group 1: n = 21 prospective longitudinal MoM hip arthroplasty; Group 2: n = 17 retrospective MoM hip arthroplasty; and Group 3: n = 20 controls without implants). We compared implant position, metal-ion release, and immuno-reactivity. MoM cohorts had elevated (p 2), compared with 76% of Group 2, and 15% of Group 3 controls (patch testing was a poor diagnostic indicator with only 1/21 Group 1 positive). Higher cup-abduction angles (50° vs. 40°) in Group 1 were associated with higher serum Cr (p < 0.07). However, sub-optimal cup-anteversion angles (9° vs. 20°) had higher serum Co (p < 0.08). Serum Cr and Co were significantly elevated in reactive versus non-reactive Group-1 participants (p < 0.04). CD4+CD69+ T-helper lymphocytes (but not CD8+) and IL-1β, IL-12, and IL-6 cytokines were all significantly elevated in metal-reactive versus non-reactive Group 1 participants. Our results showed that lymphocyte reactivity to metals can develop within the first 1-4 years after MoM arthroplasty in asymptomatic patients and lags increases in metal ion levels. This increased metal reactivity was more prevalent in those individuals with extreme cup angles and higher amounts of circulating metal.

Modern primary total hip arthroplasty femoral components have evolved to include modular necks. Subsequently, the additional taper junction provides another interface as a potential source for mechanically assisted crevice corrosion, which is a complex process involving fretting and crevice corrosion. Furthermore, it is becoming evident that an adverse local tissue reaction may result in some patients due to the mechanically assisted crevice corrosion. This article details the clinical, radiographic, and laboratory evaluation of patients with these components who present with persistent pain. The relevant surgical strategies and techniques to address this pathology in symptomatic patients are addressed.

We evaluated the consequences of cobalt-chromium alloy (CoCr) wear debris challenge in the peri-spine region to determine the inflammation and toxicity associated with submicron particulates of CoCr-alloy and nickel on the peri-spine.The lumbar epidural spaces of (n = 50) New Zealand white rabbits were challenged with: 2.5 mg CoCr, 5.0 mg CoCr, 10.0 mg CoCr, a positive control (20.0 mg of nickel) and a negative control (ISOVUE-M-300). The CoCr-alloy and Ni particles had a mean diameter of 0.2 and 0.6 μm, respectively. Five rabbits per dose group were studied at 12 and 24 weeks. Local and distant tissues were analyzed histologically and quantitatively analyzed immunohistochemically (TNF-α and IL-6).Histologically, wear particles were observed in all animals. There was no evidence of toxicity or local irritation noted during macroscopic observations in any CoCr-dosed animals. However, Ni-treated control animals experienced bilateral hind leg paralysis and were euthanized at Day 2. Histopathology of the Ni particle-treated group revealed severe neuropathy. Quantitative immunohistochemistry demonstrated a CoCr-alloy dose-dependent increase in cytokines (IL-6, TNF-α, p0.05) at 12 and 24 weeks.Subtle peri-spine inflammation associated with CoCr-alloy implant particles was dose dependent and persistent. Neuropathy can be induced by highly reactive Ni particles. This suggests peri-spine challenge with CoCr-alloy implant debris (e.g., TDA) is consistent with past reports using titanium alloy particles, i.e., mild persistent inflammation.

Innate immune reactions to orthopedic implant debris are the primary cause of total joint replacement (TJR) failure over the long term (15-20 years). The role of pathogen associated pattern recognition receptors (i.e., TLRs) in regulating immune reactivity to metal implant particles remains controversial. Do different TLRs (i.e., TLR2 vs. TLR4) activated by their respective ligands in concert with metal implant debris elicit equivalent innate immune responses? In this investigation, our in vitro and in vivo data indicate that Gram-negative PAMPs are more pro-inflammatory than Gram-positive PAMPs. In vitro results indicated TLR4 activation in concert with CoCrMo orthopedic implant debris (CoCrMo/LPS+) challenged primary macrophages resulted in significantly greater inflammatory responses than CoCrMo/PAM3CSK+ (TLR2). Similarly, in vivo results indicated CoCrMo/LPS+ TLR4 challenge induced a twofold increase in inflammation-induced bone resorption (osteolysis) than CoCrMo/PAM3CSK+ (p 0.01) or CoCrMo (p 0.03) alone in an established murine calvaria model. This points to a more potent TLR4-based effect of CoCrMo/LPS+ on innate immune responses, that is, IL-1ß, TNF-α, and resulting osteolysis. Differential CoCrMo/LPS+ induced osteolysis compared to CoCrMo/PAM3CSK+, reveals inherent differences in TLR4 versus TLR2 activation which are relevant to (i) how different types of implant debris elicit differential reactivity, (ii) how TLR2 Gram-positive bacteria benefits from less immune activation possibly due to the down-regulation of TLR2 surface expression, that subsequently impacts Gram-positive infections in TJRs, and (iii) how using TLR4 LPS (a Gram-negative agonist) may not accurately model Gram-positive bacteria responses, alone and/or with specific types of implant particles, particularly CoCrMo alloy. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:1007-1017, 2017.

Wear debris in joint replacements has been suggested as a cause of associated tissue-damaging inflammation. In this study, we examined whether solid titanium microparticles (mTi) of sufficient size to accumulate as wear debris could stimulate innate or adaptive immunity in vivo. mTi, administered in conjunction with OVA, promoted total and Ag-specific elevations in serum IgE and IgG1. Analysis of transferred transgenic OVA-specific naive T cells further showed that mTi acted as an adjuvant to drive Ag-specific Th2 cell differentiation in vivo. Assessment of the innate response indicated that mTi induced rapid recruitment and differentiation of alternatively activated macrophages in vivo, through IL-4– and TLR4-independent pathways. These studies suggest that solid microparticles alone can act as adjuvants to induce potent innate and adaptive Th2-type immune responses and further suggest that wear debris in joint replacements may have Th2-type inflammatory properties.

Study design In vitro biotribological wear particulate investigation. Objective To characterize poly-ether-ether-ketone (PEEK)-OPTIMA wear particulate generated from a series of wear tests used to evaluate the wear resistance and long-term biodurability of NUBAC, a PEEK-on-PEEK articulating nucleus replacement device, and compare with wear particulate associated with hip and knee total joint arthroplasties. Summary of background data The use of PEEK in spinal arthroplasty represents a unique application of this material. Clinically, osteolysis, osteolytic changes, and adverse reactions to metal ions have been documented in spinal arthroplasty. Therefore, it is critically important to analyze the PEEK wear particulate to evaluate its resultant biologic reactivity. Historically, scanning electron microscopy (SEM) has been used for wear debris characterization. Light scattering, specifically laser diffraction, has also been successfully used. The combined use of both techniques may provide a more comprehensive analysis than either method alone. Methods Proteinacious serum containing PEEK wear debris generated from four groups of devices from separate wear tests underwent enzymatic and acid digestion. The particulate was analyzed using laser diffraction in duplicate, followed by SEM analysis. Results Laser diffraction analysis demonstrated a relatively large mean particle diameter on the basis of particle volume (16.5-40.0 µm) as compared with particle number (0.9-2.2 µm). For all groups, more than 50% of debris was larger than 5.0 µm. SEM analysis revealed a mean particle size consistent with the number-based laser diffraction results. The morphology of the wear particulate appeared to be similar for all the groups analyzed. Conclusion The analysis of the particles generated from an articulating PEEK-on-PEEK nucleus replacement device shows debris within size ranges typical of other total joint arthroplasty implants, with relatively round morphology, along with the results suggesting a reduced particle load. These attributes tend to diminish the potential of these PEEK particles to elicit an inflammatory response.

Aseptic loosening is the devastating long term complication of total hip arthroplasty and orthopedic implant debris has been shown to trigger an intense inflammatory reaction leading to resorption of the bone matrix. Inflammatory cytokines, such as tumor necrosis factor-α (TNFα), have been implicated in this process and osteocytes may play a role in its production. We previously demonstrated that cobalt-chromium-molybdenum (CoCrMo) particles upregulate TNFα production by MLO-Y4 osteocytes in vitro, but the underlying mechanism has not been elucidated. Based on previous studies by others, we hypothesized that the calcineurin-nuclear factor of activated T cells (NFAT) pathway mediates CoCrMo particle-induced TNFα production in MLO-Y4 osteocytes. MLO-Y4 osteocytes exposed to CoCrMo particle treatment resulted in a rapid and significant increase in calcineurin activity. We also demonstrate that CoCrMo particle-induced upregulation of TNFα is reduced to control levels with calcineurin-NFAT inhibitors and this was also confirmed at mRNA level. Moreover, we demonstrate the localization of NFATs in MLO-Y4 osteocytes and that NFAT1 and 2 translocate to the nucleus upon CoCrMo particle treatment. Our results suggest that calcineurin-NFAT signaling is involved in TNFα production by MLO-Y4 osteocytes after CoCrMo particle treatment.

Wear debris-induced osteolysis is purportedly the limiting problem affecting the long term results of joint arthroplasty. Pathogenic effects of wear debris in peri-implant cells such as macrophages, osteoblasts and osteoclasts have been well studied. In contrast, the effects of wear debris on osteocytes, which make up over 90% of all bone cells, remain unknown. We hypothesized that metal implant debris can induce the pro-inflammatory response in osteocytes. This study demonstrated the effects of cobalt-chromium-molybdenum alloy (Co-Cr-Mo) particles on a well-characterized MLO-Y4 osteocyte cell line. Co-Cr-Mo alloy particle treatment significantly (p0.05) up-regulated tumor necrosis factor alpha (TNFalpha) gene expression after 3 and 6 h and TNFalpha protein production after 24 h, but down-regulated interleukin-6 (IL-6) gene expression after 6 h. Co-Cr-Mo alloy particle treatment also induced osteocyte apoptosis after 24 h. This apoptotic effect was partially (40%) dependent on TNFalpha. Therefore, our results suggest that osteocytes play a role in particle-induced inflammation and bone resorption following total joint arthroplasty by inducing pro-inflammatory cytokines and inducing osteocyte apoptosis.

Metal-on-metal bearing couples remain a popular option in total hip arthroplasty and are the only currently available option for surface replacement arthroplasty. In general, the intermediate-term clinical performance of metal-on-metal bearings has been favorable. There are, however, lingering concerns about the biologic consequences of metal release from these bearings in terms of both local tissue effects, including delayed-type hypersensitivity reactions in a subset of patients, and potential systemic effects as a consequence of chronic elevations in serum cobalt and chromium content. Advances in the understanding of the operant wear mechanisms in these bearings provide strategies for reducing the burden of metal released into the periprosthetic milieu, which in turn will mitigate the concerns about the biologic response to the metal debris. Continued surveillance of patients with these bearings is warranted to determine whether metal-on-metal bearing couples provide a long-term survivorship advantage over other bearing couple options and to evaluate whether chronic elevations in the body burden of cobalt and chromium is well tolerated over the long term.

Immune reactivity to soluble and particulate implant debris remains the primary cause of aseptic inflammation and implant loosening. However, the intracellular mechanisms that trigger immune cells to sense and respond to exogenous nonbiological agents such as metal particles or metal ions released from orthopedic implants remain unknown. Recent studies in immunology have outlined the importance of the intracellular inflammasome complex of proteins in sensing danger/stress signals triggered by nonbiological agents in the cytosol of macrophages. We hypothesized that metal implant debris can activate the inflammasome pathway in macrophages that causes caspase-1-induced cleavage of intracellular pro-IL-1β into its mature form, resulting in IL-1β secretion and induction of a broader proinflammatory response. We tested this hypothesis by examining whether soluble cobalt, chromium, molybdenum, and nickel ions and Co-Cr-Mo alloy particles induce inflammasome- mediated macrophage reactivity. Our results demonstrate that these agents stimulate IL-1β secretion in human macrophages that is inflammasome mediated (i.e., NADPH-, caspase-1-, Nalp3-, and ASC-dependent). Thus, metal ion- and particle-induced activation of the inflammasome in human macrophages provides evidence of a novel pathway of implant debris-induced inflammation, where contact with implant debris is sensed and transduced by macrophages into a proinflammatory response. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27: 847–854, 2009

Background The increasing complexity of articulating spinal implants prohibits the use of serum-supplemented simulator fluid testing because multicomponent interfaces retain residual protein and preclude gravimetric measurement. Our original hypothesis was that simulator testing of a posterior dynamic stabilization implant that has metal-on-metal articulating bearings will not produce dramatically different wear debris when tested using pure saline versus testing in saline supplemented with 20% serum. Methods This hypothesis was tested using simulator testing of 12 dynamic stabilization spinal implants, 6 in 100% saline and 6 in 20%-serum saline. Gravimetric and particle analysis were performed after every million cycles up to 10 million cycles, with flexion of 11.3°/extension of 5.6° coupled with axial rotation of ± 4°. Results The mean gravimetric weight loss was approximately 200 mg over 10 million cycles for the implants tested in 100% saline, while the mean weight loss for those tested in 20%-serum saline was below the method detection limits (< 10 mg over 10 million cycles). For the 100%-saline and 20%-serum simulator fluids, the average particle size over the course of 0 to 10 million cycles remained relatively constant at 0.2 µm-dia (saline) and 3.2 µm-dia (20%-serum saline). Testing in 100% saline generated > 1000-fold more particles, compared to testing in 20% serum-supplemented saline. Energy-dispersive X-ray (EDAX) analyses of particles demonstrated that the 100% saline debris was composed of Co-Cr-P-O (Cr-Co metal oxides), and for the 20%-serum saline debris only bulk metal Co-Cr was detected. Conclusion Our initial hypothesis was not supported. There were significant differences in gravimetric wear, average size, and type of wear debris that were mechanistically attributable to the type of simulator fluid used. The over-protective effect of serum proteins appears to underscore the importance of using both saline and serum when establishing upper and lower bounds of predictive implant debris generation modeling, where saline represents a worst-case scenario and as little as 20% serum masks all weight loss completely in highly modular articulating implants. Clinical Relevance Clinical Relevance = 5 (Oxford Centre for Evidence-based Medicine Levels of Evidence). Study findings are limited to a greater understanding of the science associated with predictive wear testing of articulating spinal implants.

It remains unclear how metal released from implants affects cells of the immune system and, in particular, cells of the adaptive immune system, that is, T-helper lymphocytes. In this study, we investigated the effects of aluminum, chromium, cobalt, copper, iron, molybdenum, nickel, niobium, vanadium, and zirconium ions at concentrations from 0.05 to 5.0 mM on human CD4+ T lymphocytes. The DNA damage, apoptosis, necrosis, and proliferation responses of a human T-helper lymphocyte (Jurkat) cell line were evaluated to test our hypothesis that some metals will preferentially induce genotoxicity (DNA damage). Our results demonstrated that metal ions did not preferentially induce Jurkat T-lymphocyte DNA damage prior to other forms of toxicity, that is, apoptosis and/or direct necrosis. Nickel and vanadium induced the most DNA damage and were the most apoptotic metals tested, inducing >50% caspase-9 positive T cells at 0.05 mM and 0.1 mM concentrations, respectively. Cobalt and niobium were the most toxic metals, inducing 50% apoptosis (i.e.

STUDY DESIGN: The study is a prospective observational study of 48 continuous patients with symptomatic lumbar degenerative disk disease. Each patient underwent discography, MRI, and a biochemical analysis of disk lavage fluid. OBJECTIVES: The purpose of this study was to correlate concordant pain on discography with MRI grade and biochemical markers of inflammation in a clinical setting. SUMMARY OF BACKGROUND DATA: The pathophysiology of degenerative disk disease is complex. Discography is used to differentiate symptomatic from asymptomatic levels. MRI is used to image changes in disk water content. Biochemical assays have identified molecular markers of inflammation. To date, no study has correlated concordant pain on discography with MRI findings and biochemical markers. METHODS: Forty-eight (48) continuous patients with symptomatic lumbar degenerative disk disease gave informed consent for study entry. Patient sex, age, insurance, work status and visual analog score (VAS) were recorded. MRI was obtained and Pfirrmann grading was performed by a single spine surgeon. Discography with disc lavage was performed by a single anesthesiologist. Lavage samples were tested for inflammatory markers with high resolution multi-plex bead immunoassays and ELISA with O5 pg/ml resolution. RESULTS: None of demographic variables was significantly related to concordant pain on discogram by chi-squared tests and Mann-Whitney U-test. The Pfirrmann score was significantly different for patients with and without concordant pain at L3-L4 (p!0.001), but was insignificant at other levels after multitest correction. Pfirrmann scores were significantly different at any level in patients with and without concordant pain. VAS scores were not significantly correlated with opening pressures at any level. Despite the presence of serum proteins in the disk lavage fluid, none of the tested inflammatory mediators was identified by multi-plex bead immunoassays and ELISA. CONCLUSIONS: There are only weak correlations between demographic, discogram, and radiographic variables. Response to discogram cannot be predicted by non-invasive means. The disk lavage method was unable to identify the presence of specific inflammatory peptides with multiplex immunoassays and ELISA. 2008 Elsevier Inc. All rights reserved.