A novel method for isolation and recovery of ceramic nanoparticles and metal wear debris from serum lubricants at ultra-low wear rates

Lal, Saurabh; Hall, Richard M.; Tipper, Joanne L.

doi:10.1016/j.actbio.2016.07.004

Cited by 15 publications

(29 citation statements)

References 42 publications

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“…Proteins were effectively removed and the isolated particles were relatively pure, and were not irreversibly bound to any surface, so particles isolated using the method have the potential to be used in further studies; for example with nanoparticle-tracking analysis. This study has taken the initial progress made by [28] forward by adapting the isolation process for a new application -the isolation of particles from tissue samples. This has involved changes to the method presented by [28] including sample preparation, the removal of certain stages of the protocol, and optimisation of the digestion protocol specifically for fixed tissue samples.…”

Section: Discussionmentioning

confidence: 99%

“…This study has taken the initial progress made by [28] forward by adapting the isolation process for a new application -the isolation of particles from tissue samples. This has involved changes to the method presented by [28] including sample preparation, the removal of certain stages of the protocol, and optimisation of the digestion protocol specifically for fixed tissue samples. Previously the isolation process had not been tested on micron-scale particles, only nanoscale particles, and had also not been used to isolate titanium particles.…”

Section: Discussionmentioning

confidence: 99%

“…The recovery rate of the particle isolation method detailed in [28], which utilised sodium polytungstate density gradient ultracentrifugation to isolate particles from wear simulator serum, was tested gravimetrically by weighing recovered particles on filters as detailed in [32] to verify the efficacy of using sodium polytungstate gradients. A novel tissue digestion method was developed and optimised as described in [33] to produce an isolation protocol for tissue samples.…”

Section: Particle Isolationmentioning

confidence: 99%

“…The method described by [28] to isolate Si 3 N 4 particles from wear simulator serum using density gradient ultracentrifugation with novel sodium polytungstate gradients demonstrated no detectable particle loss at key stages of the procedure, and no detectable changes to particle size or morphology. The method is relatively quick to perform (the protocol requires approximately four days), is cost effective and does not require specialist equipment, making it an attractive technique.…”

Section: Introductionmentioning

confidence: 99%

“…Although several recent publications have provided updated methods for isolating low-wearing materials from serum with proven sensitivity [27,28], such studies have not considered the isolation of particles from tissue, which can be useful when performed on periprosthetic tissue from around explants for diagnostic purposes. The method described by [28] to isolate Si 3 N 4 particles from wear simulator serum using density gradient ultracentrifugation with novel sodium polytungstate gradients demonstrated no detectable particle loss at key stages of the procedure, and no detectable changes to particle size or morphology.…”

Section: Introductionmentioning

confidence: 99%

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Recovery of low volumes of wear debris from rat stifle joint tissues using a novel particle isolation method

et al. 2018

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a b s t r a c t Less than optimal particle isolation techniques have impeded analysis of orthopaedic wear debris in vivo. The purpose of this research was to develop and test an improved method for particle isolation from tissue. A volume of 0.018 mm 3 of clinically relevant CoCrMo, Ti-6Al-4V or Si 3 N 4 particles was injected into rat stifle joints for seven days of in vivo exposure. Following sacrifice, particles were located within tissues using histology. The particles were recovered by enzymatic digestion of periarticular tissue with papain and proteinase K, followed by ultracentrifugation using a sodium polytungstate density gradient. Particles were recovered from all samples, observed using SEM and the particle composition was verified using EDX, which demonstrated that all isolated particles were free from contamination. Particle size, aspect ratio and circularity were measured using image analysis software. There were no significant changes to the measured parameters of CoCrMo or Si 3 N 4 particles before and after the recovery process (KS tests, p > 0.05). Titanium particles were too few before and after isolation to analyse statistically, though size and morphologies were similar. Overall the method demonstrated a significant improvement to current particle isolation methods from tissue in terms of sensitivity and efficacy at removal of protein, and has the potential to be used for the isolation of ultra-low wearing total joint replacement materials from periprosthetic tissues. Statement of SignificanceThis research presents a novel method for the isolation of wear particles from tissue. Methodology outlined in this work would be a valuable resource for future researchers wishing to isolate particles from tissues, either as part of preclinical testing, or from explants from patients for diagnostic purposes. It is increasingly recognised that analysis of wear particles is critical to evaluating the safety of an orthopaedic device.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Particle Isolationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Recovery of low volumes of wear debris from rat stifle joint tissues using a novel particle isolation method

et al. 2018

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Quantitative analysis and degradation mechanisms of different protein degradation methods

et al. 2021

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The abrasive debris produced by wear test of artificial joints in vitro is encapsulated by proteins in serum lubricants, which hinder the characterization of debris analysis.One of the key issues of isolating wear debris from serum is degrading the proteins wrapping the wear debris. In this article, the proteins in calf serum were degraded by a strong alkali, a strong acid, and an enzyme. The residual concentration of proteins in calf serum was detected by UV absorption. Quantitative analysis of protein degradation and the protein degradation rate was proposed, following treatment with different degradation reagents and different incubation times. The results showed that when 10 mL of 25% volume calf serum was added with 40 mL of NaOH and incubated at 65 C for 24 h, the protein degradation rate reached a maximum of 95.52%.The protein degradation rate in the solution ranged from 31.86% to 71.64% when a different volume of 37% HCl was added and incubated at 60 C. The highest protein degradation rate was 94.98% in the protease degradation solution. When the protein degradation rate is less than 70%, the particles were coated by protein. When the protein degradation rate was more than 95%, there was no protein coating around the particles. The three protein degradation methods have different processes and protein degradation rates. A suitable method for protein degradation can be selected according to these practical applications.artificial joints, degradation rate, protein degradation, recovery of debris | INTRODUCTIONAfter an artificial joint is implanted into the human body, it is not only soaked by body fluids but is also impacted by and subjected to normal and shear forces. The abrasive particles generated by the wear of joints induce osteoclasts to produce osteolytic factors and induce bone resorption. Aseptic loosening of the prosthesis due to bone dissolution is the most common cause of the failure of the prosthesis. [1][2][3] The different sizes and morphologies of debris reflect the different wear behavior and mechanism of the artificial joint in addition to the different degrees of biological reaction. [4][5][6][7][8] Therefore, a reliable method of isolating debris from serum during artificial joint wear tests in vitro is important to the analysis of debris characteristics.Calf serum has been widely used as the lubricating fluid in vitro for wear tests of artificial joints. [9][10][11] Artificial joint debris binds to proteins in serum by ionization, hydrophobicity, and hydrophilicity. The joint debris is wrapped by multiple layers of proteins called the protein halo. 12-14 Therefore, the degree of protein degradation affects the separation and analysis of the amount and morphology of the debris.A suitable protein degradation method is an important prerequisite for the isolation and analysis of wear debris.Many scholars have researched different protein degradation methods. Different protein degradation methods have different requirements for the amount of strong acid, strong alkali and protease, temperat...

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Mixed material wear particle isolation from periprosthetic tissue surrounding total joint replacements

et al. 2022

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Submicron-sized wear particles are generally accepted as a potential cause of aseptic loosening when produced in sufficient volumes. With the accelerating use of increasingly wear-resistant biomaterials, identifying such particles and evaluating their biological response is becoming more challenging. Highly sensitive wear particle isolation methods have been developed but these methods cannot isolate the complete spectrum of particle types present in individual tissue samples. Two established techniques were modified to create one novel method to isolate both high-and low-density materials from periprosthetic tissue samples. Ten total hip replacement and eight total knee replacement tissue samples were processed. All particle types were characterized using high resolution scanning electron microscopy. UHMWPE and a range of high-density materials were isolated from all tissue samples, including: polymethylmethacrylate, zirconium dioxide, titanium alloy, cobalt chromium alloy and stainless steel. This feasibility study demonstrates the coexistence of mixed particle types in periprosthetic tissues and provides researchers with high-resolution images of clinically relevant wear particles that could be used as a reference for future in vitro biological response studies.

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A novel method for isolation and recovery of ceramic nanoparticles and metal wear debris from serum lubricants at ultra-low wear rates

Cited by 15 publications

References 42 publications

Recovery of low volumes of wear debris from rat stifle joint tissues using a novel particle isolation method

Recovery of low volumes of wear debris from rat stifle joint tissues using a novel particle isolation method

Quantitative analysis and degradation mechanisms of different protein degradation methods

Mixed material wear particle isolation from periprosthetic tissue surrounding total joint replacements

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