Vivo Oxidation Research Articles

In Vivo Oxidation and Wear in Remelted Highly Cross-Linked Polyethylene Liners Retrieved at a Minimum of 10 Years After Total Hip Arthroplasty

BackgroundWith the decreasing age threshold for patients undergoing total hip arthroplasty (THA), there is an escalating demand for enhanced polyethylene durability. Although reports assessing wear in remelted highly cross-linked polyethylene (HXLPE) through radiographic imaging exist, a consensus regarding its oxidation level is lacking. This study investigated the wear, oxidation levels, and degradation of remelted HXLPE that was retrieved at least 10 years after THA. MethodsOur analysis focused on 7 cases of melted HXLPE liners retrieved ≥10 years after THA. All patients were women, who had an average age of 64 ± 6.5 years at the initial operation, and the mean postoperative period after THA was 12 years and 11 months ±1 year and 5 months. The wear conditions were measured by matching the shape analysis data obtained from a coordinate-measuring machine with a spherical model. Fourier-transform infrared spectroscopy was used to study the oxidation of polyethylene, and the polyethylene structure was evaluated using scanning electron microscopy. ResultsOsteolysis was not observed in any case on X-rays, computed tomography, or intraoperative findings during revision surgery. The average oxidation index (OI) of the sliding surface under load was 0.31 ± 0.22 in the 6 cases calculated after hexane treatment, and 0.69 for one case without hexane treatment. In the 6 cases calculated after hexane treatment, the average OI of the non–load-bearing sliding surface was 0.11 ± 0.20. Average wear values were 0.33 ± 0.11 mm at 45 degrees from the equatorial direction and 0.04 ± 0.07 mm in the opposite direction. The initial structure of the polyethylene was preserved at all sites with low oxidation levels; however, in one case with stem subsidence, morphological changes and a high OI were observed. ConclusionsLong-term oxidation and wear of remelted HXLPE liners retrieved from THA patients were minimal.

Don't Throw the Baby Out with the Bathwater: Commentary on an article by Barbara H. Currier, MChE, et al.: "Oxidation in Retrieved, Never-Irradiated UHMWPE Bearings. What Can We Learn About in Vivo Oxidation?"

Don't Throw the Baby Out with the Bathwater: Commentary on an article by Barbara H. Currier, MChE, et al.: "Oxidation in Retrieved, Never-Irradiated UHMWPE Bearings. What Can We Learn About in Vivo Oxidation?"

In Vivo Oxidation and Surface Damage in Retrieved Ethylene Oxide-sterilized Total Knee Arthroplasties

Gas sterilization (eg, ethylene oxide [EtO] and gas plasma) was introduced for polyethylene to reduce oxidation due to free radicals occurring during radiation sterilization. Recently, oxidation has been observed in polyethylenes with undetectable levels of free radicals, which were expected to be oxidatively stable. It is unclear whether in vivo oxidation will occur in unirradiated inserts sterilized with EtO. We analyzed the oxidation, mechanical behavior, and surface damage mechanisms of tibial inserts of a single design sterilized using EtO. We collected 20 EtO-sterilized tibial inserts at revision surgeries. We assessed oxidative using Fourier transform infrared spectroscopy and mechanical properties using the small punch test. Surface damage was assessed using damage scoring techniques and micro-CT. Oxidation indexes were low and uniform between the regions. The subtle changes did not affect the mechanical properties of the polymer. The dominant surface damage modes included burnishing, abrasion, and third-body wear. There was no evidence of delamination in the retrievals. The retrieved EtO-sterilized UHMWPE retrievals remained stable with respect to both oxidative and mechanical properties for up to 10 years in vivo. We did observe slight measurable amounts of oxidation in the inserts; however, it was far below levels that would be expected to compromise the strength of the polymer. Due to the stable oxidative and mechanical properties, EtO-sterilized tibial components appear to be an effective alternative to gamma-sterilized inserts, at least in short-term implantations.

In Vivo Oxidation in Remelted Highly Cross-Linked Retrievals

Elimination of free radicals to prevent oxidation has played a major role in the development and product differentiation of the latest generation of highly cross-linked ultra-high molecular weight polyethylene bearing materials. In the current study, we (1) examined oxidation in a series of retrieved remelted highly cross-linked ultra-high molecular weight polyethylene bearings from a number of device manufacturers and (2) compared the retrieval results with findings for shelf-stored control specimens. The hypothesis was that radiation-cross-linked remelted ultra-high molecular weight polyethylene would maintain oxidative stability in vivo comparable with the stability during shelf storage and in published laboratory aging tests. Fifty remelted highly cross-linked ultra-high molecular weight polyethylene acetabular liners and nineteen remelted highly cross-linked ultra-high molecular weight polyethylene tibial inserts were received after retrieval from twenty-one surgeons from across the U.S. Thirty-two of the retrievals had been in vivo for two years or more. Each was measured for oxidation with use of Fourier transform infrared spectroscopy. A control series of remelted highly cross-linked ultra-high molecular weight polyethylene acetabular liners from three manufacturers was analyzed with electron paramagnetic resonance spectroscopy to measure free radical content and with Fourier transform infrared spectroscopy to measure oxidation initially and after eight to nine years of shelf storage in air. The never-implanted, shelf-aged controls had no measurable free-radical content initially or after eight to nine years of shelf storage. The never-implanted controls showed no increase in oxidation during shelf storage. Oxidation measurements showed measurable oxidation in 22% of the retrieved remelted highly cross-linked liners and inserts after an average of two years in vivo. Because never-implanted remelted highly cross-linked ultra-high molecular weight polyethylene materials had no measurable free-radical concentration and no increase in oxidation during shelf storage, these materials were expected to be oxidation-resistant in vivo. However, some remelted highly cross-linked ultra-high molecular weight polyethylene retrievals showed measurable oxidation after an average of more than two years in vivo. This apparent departure from widely expected behavior requires continued study of the process of in vivo oxidation of ultra-high molecular weight polyethylene materials.

Commentary on an Article by B.H. Currier, MChE, et al.: “In Vivo Oxidation in Remelted Highly Cross-Linked Retrievals”

Total joint arthroplasty is a highly successful procedure; however, the one major issue that remains is wear of the bearing surfaces and the resultant adverse biological response to wear particles. Many studies of ultra-high molecular weight polyethylene have indicated that residual free radicals combined with oxygen result in increased brittleness and reduction of mechanical properties. The development of highly cross-linked ultra-high molecular weight polyethylene with use of irradiation in inert environments, and subsequent free-radical stabilization by either annealing or remelting, provided a bearing surface with the potential for significant reduction of wear. There have been several clinical studies that have confirmed, with use of radiographic techniques, this reduction in wear. However, in the recent radiographic study by Currier et al., an unexpected increase in wear was seen in this ultra-high molecular weight polyethylene after seven years in vivo. This troubling observation requires further study. Currier …

In Vivo Oxidation Contributes to Delamination but not Pitting in Polyethylene Components for Total Knee Arthroplasty

The aim of this study was to better understand how in vivo oxidation contributes to fatigue damage in total knee arthroplasty (TKA). A total of 119 tibial inserts were consecutively collected after revision surgery. Of the 119 polyethylene retrievals, 29 were gamma sterilized in air (historical), whereas the remaining 90 were gamma sterilized in nitrogen (conventional). Surface damage assessment and characterization of oxidation were performed on all the retrievals. Delamination was significantly more prevalent and extensive in the longer-term, highly oxidized, historical tibial inserts. Pitting damage, in contrast, seemed to be equally prevalent between both retrieval groups and was not correlated with in vivo oxidation. Our findings support our hypothesis that in vivo oxidation is a contributing factor to delamination, but not pitting, in TKA. Despite the lower oxidation displayed by conventional retrievals, this study provides strong evidence that delamination secondary to in vivo oxidation may occur during the second decade of implantation.

Identification of the Intermediates of in Vivo Oxidation of 1,4-Dioxane by Monooxygenase-Containing Bacteria

1,4-dioxane is a probable human carcinogen and an emerging water contaminant. Monooxygenase-expressing bacteria have been shown to degrade dioxane via growth-supporting as well as cometabolic mechanisms. In this study, the intermediates of dioxane degradation by monooxygenase-expressing bacteria were determined by triple quadrupole-mass spectrometry and Fourier transform ion cyclotron resonance-mass spectrometry. The major intermediates were identified as 2-hydroxyethoxyacetic acid (HEAA), ethylene glycol, glycolate, and oxalate. Studies with uniformly labeled 14C dioxane showed that over 50% of the dioxane was mineralized to CO2 by CB1190, while 5% became biomass-associated after 48 h. Volatile organic acids and non-volatiles, respectively, accounted for 20 and 11% of the radiolabeled carbon. Although strains cometabolizing dioxane exhibited limited transformation capacities, nearly half of the initial dioxane was recovered as CO2. On the basis of these analytical results, we propose a pathway for dioxane oxidation by monooxygenase-expressing cells in which dioxane is first converted to 2-hydroxy-1,4-dioxane, which is spontaneously oxidized to HEAA. During a second monooxygenation step, HEAA is further hydroxylated, resulting in a mixture of dihydroxyethoxyacetic acids with a hydroxyl group at the ortho or para position. After cleavage of the second ether bond, small organic molecules such as ethylene glycol, glycolate, glyoxalate, and oxalate are progressively formed, which are then mineralized to CO2 via common cellular metabolic pathways. Bioremediation of dioxane via this pathway is not expected to cause an accumulation of toxic compounds in the environment.

In Vivo Oxidation of γ-Barrier–Sterilized Ultra–High-Molecular-Weight Polyethylene Bearings

γ-Barrier packaging is shown to be effective in preventing oxidation of polyethylene during shelf storage and in addressing the problem of early fatigue failure seen in γ-air–sterilized bearings with long shelf-storage before implantation. The series of γ-barrier retrievals studied suggests that oxidation occurs in the body via the same mechanism as seen in γ-air–sterilized bearings. A critical oxidation level is identified above which polyethylene bearings are susceptible to fatigue damage after sufficient cycles of use. Although critical oxidation was not reached in the majority of the retrieved γ-barrier bearings studied, in vivo oxidation appears to follow an exponential increase with time. The result of in vivo oxidation is expected to be loss of mechanical properties, and susceptibility of polyethylene bearings to eventual fatigue failure.

In Vivo Oxidation of Retrieved Cross-linked Ultra–High-Molecular-Weight Polyethylene Acetabular Components with Residual Free Radicals

Wear of ultra–high-molecular-weight polyethylene (UHMWPE) contributes to debris that can lead to periprosthetic osteolysis in total hip arthroplasty. Irradiation not only decreases wear of UHMWPE but also generates residual free radicals that can oxidize the UHMWPE in the long term. Melting or annealing is used to quench the free radicals. Melting is more effective than annealing. We hypothesized that the postirradiation annealed UHMWPE components would oxidize in vivo and that postirradiation melted ones would not. We analyzed surgical explants of UHMWPE acetabular liners. The irradiated and annealed explants showed embrittlement, oxidation, and an increase in crystallinity. The irradiated and melted UHMWPE explants showed no oxidation, no increase in crystallinity, and no embrittlement. To prevent long-term chemical changes in highly cross-linked UHMWPE components, the residual free radicals must be stabilized after irradiation, preferably by melting and not annealing.

Changes in the expression and the enzymic properties of the 20S proteasome in sugar-starved maize roots. evidence for an in vivo oxidation of the proteasome.

The 20S proteasome (multicatalytic proteinase) was purified from maize (Zea mays L. cv DEA 1992) roots through a five-step procedure. After biochemical characterization, it was shown to be similar to most eukaryotic proteasomes. We investigated the involvement of the 20S proteasome in the response to carbon starvation in excised maize root tips. Using polyclonal antibodies, we showed that the amount of proteasome increased in 24-h-carbon-starved root tips compared with freshly excised tips, whereas the mRNA levels of alpha 3 and beta 6 subunits of 20S proteasome decreased. Moreover, in carbon-starved tissues, chymotrypsin-like and caseinolytic activities of the 20S proteasome were found to increase, whereas trypsin-like activities decreased. The measurement of specific activities and kinetic parameters of 20S proteasome purified from 24-h-starved root tips suggested that it was subjected to posttranslational modifications. Using dinitrophenylhydrazine, a carbonyl-specific reagent, we observed an increase in carbonyl residues in 20S proteasome purified from starved root tips. This means that 20S proteasome was oxidized during starvation treatment. Moreover, an in vitro mild oxidative treatment of 20S proteasome from non-starved material resulted in the activation of chymotrypsin-like, peptidyl-glutamyl-peptide hydrolase and caseinolytic-specific activities and in the inhibition of trypsin-like specific activities, similar to that observed for proteasome from starved root tips. Our results provide the first evidence, to our knowledge, for an in vivo carbonylation of the 20S proteasome. They suggest that sugar deprivation induces an oxidative stress, and that oxidized 20S proteasome could be associated to the degradation of oxidatively damaged proteins in carbon starvation situations.

Infection and inflammation induce LDL oxidation in vivo.

Epidemiological studies have shown an increased incidence of coronary artery disease in patients with chronic infections and inflammatory disorders. Because oxidative modification of lipoproteins plays a major role in atherosclerosis, the present study was designed to test the hypothesis that the host response to infection and inflammation induces lipoprotein oxidation in vivo. Lipoprotein oxidation was measured in 3 distinct models of infection and inflammation. Syrian hamsters were injected with bacterial lipopolysaccharide (LPS), zymosan, or turpentine to mimic acute infection, acute systemic inflammation, and acute localized inflammation, respectively. Levels of oxidized fatty acids in serum and lipoprotein fractions were measured by determining levels of conjugated dienes, thiobarbituric acid-reactive substances, and lipid hydroperoxides. Our results demonstrate a significant increase in conjugated dienes and thiobarbituric acid-reactive substances in serum in all 3 models. Moreover, LPS and zymosan produced a 4-fold to 6-fold increase in conjugated diene and lipid hydroperoxide levels in LDL fraction. LPS also produced a 17-fold increase in LDL content of lysophosphatidylcholine that is formed during the oxidative modification of LDL. Finally, LDL isolated from animals treated with LPS was significantly more susceptible to ex vivo oxidation with copper than LDL isolated from saline-treated animals, and a 3-fold decrease occurred in the lag phase of oxidation. These results demonstrate that the host response to infection and inflammation increases oxidized lipids in serum and induces LDL oxidation in vivo. Increased LDL oxidation during infection and inflammation may promote atherogenesis and could be a mechanism for increased incidence of coronary artery disease in patients with chronic infections and inflammatory disorders.

In Vivo Oxidation of [13C]Galactose in Patients with Galactose-1-Phosphate Uridyltransferase Deficiency

We developed an intravenous and oral [13C]galactose breath test for the in vivo study of galactose metabolism. Following an intravenous bolus of 7 mg/kg of [1-13C]galactose in the fasting state, normal children and adults eliminated 3-6% and 21-47% of the bolus as 13CO2 in expired air collected over 1 and 5 h, respectively, Comparable fractional elimination was seen when the dose was given orally. Patients with galactosemia who have barely detectable or absent galactose-l-phosphate uridyltransferase (GALT) activity in erythrocytes and are homoallelic for the Q188R gene mutation, when given a 7 mg/kg intravenous bolus had barely detectable 13CO2 in air samples in the first hour, but eventually eliminated as much as 3.6% of the dose in 5 h. A galactosemia/Duarte (Q188R/N314D) compound heterozygote and a homozygous Duarte subject, as well as a subject with one normal allele and one Q188R allele, showed normal in vivo oxidation. An assessment of whole body galactose metabolism can be made with this procedure. Further use of this in vivo modality in patients with different genetic backgrounds should increase our understanding of genotype-phenotype relationships in hereditary galactosemia.

In Vivo Oxidation of Ronnel in the Madeira Cockroach1

The oxygen analog of ronnel, O,O -dimethyl O -(2,4,5-trichlorophenyl) phosphate, was identified in the extracts of ronnel-treated Madeira cockroaches, Leucophaea maderae (F.). The extracts were analyzed by cholinesterase assay for inhibitors on intact paper chromatograms developed by normal and reverse phase solvent systems. It was established that the phosphate was produced by in vivo oxidation of ronnel, O,O -dimethyl O -(2,4,5-trichlorophenyl) phosphorothioate, in the cockroaches and not during subsequent analytical steps or by administration of the phosphate as a contaminant of technical ronnel. The enzyme method is highly sensitive, easily detecting 0.01 µg. or less of the phosphate and when combined with several chromatographic systems should be widely applicable in detecting and identifying anticholinesterase metabolites formed in biological systems.