Through-thickness patterns of shear strain evolve in early osteoarthritis.

OBJECTIVE: Given the structural changes associated with the progression of Osteoarthritis (OA), we hypothesized that patterns of through-thickness, large-strain shear evolve with early-stage OA. We therefore aimed to determine whether and how patterns of shear strains change during early-stage OA to 1) gain insight into the progression of OA by quantifying changes in local deformations; 2) gauge the potential of patterns in shear strain to serve as image-based biomarkers of early-stage OA; and 3) provide high-resolution, through-thickness data for proposing, fitting, and validating constitutive models for cartilage. DESIGN: We completed displacement-driven, large-strain shear tests (5, 10, 15%) on 44 specimens of variably advanced osteoarthritic human articular cartilage as determined by both Osteoarthritis Research Society International (OARSI) grade and PLM-CO score. We recorded the through-thickness deformations with a stereo-camera system and processed these data using digital image correlation (DIC) to determine full-thickness patterns of strains and relative zonal recruitments, i.e., the average shear strain in a through-thickness zone weighted by its relative thickness and normalized by the applied strain. RESULTS: We observed three general shapes for the curves of averaged through-thickness, Green-Lagrange shear strains during progression of OA. We also observed that during the progression of OA only the deep zone is recruited differently under shear in a statistically significant way. CONCLUSIONS: We propose that changes in through-thickness patterns of shear strain could provide sensitive biomarkers for early clinical detection of OA. The relative zonal recruitment of the deep zone decreases with progressing OA (OARSI grade) and microstructural remodeling (PLM-CO score), which do not consistently affect recruitment of the superficial and middle zones.

The evolving large-strain shear responses of progressively osteoarthritic human cartilage.

OBJECTIVE: The composition and structure of articular cartilage evolves during the development and progression of osteoarthritis (OA) resulting in changing mechanical responses. We aimed to assess the evolution of the intrinsic, large-strain mechanics of human articular cartilage-governed by collagen and proteoglycan and their interactions-during the progression of OA. DESIGN: We completed quasi-static, large-strain shear tests on 64 specimens from ten donors undergoing total knee arthroplasty (TKA), and quantified the corresponding state of OA (OARSI grade), structural integrity (PLM score), and composition (glycosaminoglycan and collagen content). RESULTS: We observed nonlinear stress-strain relationships with distinct hystereses for all magnitudes of applied strain where stiffnesses, nonlinearities, and hystereses all reduced as OA advanced. We found a reduction in energy dissipation density up to 80% in severely degenerated (OARSI grade 4, OA-4) vs normal (OA-1) cartilage, and more importantly, we found that even cartilage with a normal appearance in structure and composition (OA-1) dissipated 50% less energy than healthy (control) load-bearing cartilage (HL0). Changes in stresses and stiffnesses were in general less pronounced and did not allow us to distinguish between healthy load-bearing controls and very early-stage OA (OA-1), or to distinguish consistently among different levels of degeneration, i.e., OARSI grades. CONCLUSIONS: Our results suggest that reductions in energy dissipation density can be detected by bulk-tissue testing, and that these reductions precede visible signs of degeneration. We highlight the potential of energy dissipation, as opposed to stress- or stiffness-based measures, as a marker to diagnose early-stage OA.

Aspirin reduces blood cholesterol in copper-deficient rats: a potential antioxidant agent?

The purpose of this study was to examine whether the hypocholesterolemic effect of aspirin is to due to its antioxidant properties. Oxidative stress was induced in rats by feeding them a copper-deficient diet. Copper deficiency reduced the activity of the enzyme superoxide dismutase (SOD) and lowered liver copper concentration but elevated liver iron. The combination of reduced SOD activity, high liver iron, and low liver copper resulted in an oxidative stress assessed by increased liver lipid peroxidation compared with copper-adequate controls. In addition, copper-deficient rats exhibited elevation of blood cholesterol. The administration of aspirin lowered both liver lipid peroxidation and blood cholesterol. It is suggested that the hypocholesterolemic properties of aspirin could be due to its ability to reduce oxidative stress.

Intestinal beta-carotene 15,15'-dioxygenase activity is markedly enhanced in copper-deficient rats fed on high-iron diets and fructose.

The purpose of the present work was to examine effects of the Cu-Fe interaction on intestinal beta-carotene 15,15'-dioxygenase activity when a wide range of dietary Fe (deficiency to excess) was used in relation to Cu status of rats. The effect of dietary carbohydrates was also examined since they play a role in the Cu-Fe interaction in vivo. Weanling male Sprague-Dawley rats (n 72) were divided into twelve dietary groups, which were fed on either low-, normal-, or high-Fe levels (0.9, 9.0, and 90.0 mmol Fe/kg diet respectively) combined with Cu-adequate or -deficient levels (0.94 and 0.09 mmol Cu/kg diet respectively) and with starch or fructose in the diets. The data showed that both Fe concentration and beta-carotene 15,15'-dioxygenase activity in small intestinal mucosa were enhanced with increasing dietary Fe and with Cu deficiency v. Cu adequacy. Dietary fructose did not aggravate the Fe-enhancement, related to Cu deficiency, in the small intestine; however, fructose increased the intestinal dioxygenase activity in rats fed on normal- or high-Fe diets when compared with starch controls. Thus, the highest intestinal dioxygenase activity associated with the lowest hepatic retinol (total) concentration was found in rats fed on the Cu-deficient, high-Fe, fructose-based diet. Finally, a positive linear relationship was found between the dioxygenase activity and Fe concentration in intestinal mucosa. In conclusion, the data indicate that beta-carotene 15,15'-dioxygenase activity requires Fe as cofactor in vivo and the enzyme is modulated by the three dietary components: Cu, Fe, and fructose.

Dietary fructose but not starch is responsible for hyperlipidemia associated with copper deficiency in rats: effect of high-fat diet.

OBJECTIVE: To test the hypothesis that copper deficiency in rats may be hyperlipidemic only when the diets consumed contain nutrients which contribute to blood lipids such as fructose and high fat. METHODS: Weanling male Sprague Dawley rats were fed diets which contained either starch or fructose as their sole carbohydrate source. The diets were either inadequate (0.6 microg Cu/g) or adequate (6.0 microg Cu/g) in copper and contained either high (300 g/kg) or low (60 g/kg) fat. At the end of the 4th week the rats were killed. Livers were analyzed for copper content. Plasma was analyzed for cholesterol and triglyceride concentrations. RESULTS: High-fat diet did not increase blood lipids in rats fed a copper-deficient diet containing starch. In contrast, the combination of high-fat diet with fructose increased blood triglycerides and fructose with copper deficiency resulted in a significant increases in blood cholesterol. CONCLUSIONS: Hyperlipidemia of copper deficiency in rats is dependent on synergistic effects between dietary fructose and copper deficiency and fructose and amount of dietary fat. Hyperlipidemia does not develop if starch is the main source of dietary carbohydrate in a copper-deficient diet even if a high-fat diet is fed.

Cholesterol-lowering nature of unsaturated fat in rats may be due to its inability to increase hepatic iron.

The present investigation was conducted to determine whether the cholesterol-raising properties of saturated fat and cholesterol-lowering properties of unsaturated fat are associated with levels of hepatic iron. The magnitude of hepatic iron retention was manipulated by feeding rats diets that were either copper-deficient or -adequate, iron-adequate or -supplemented, and contained either beef tallow or corn oil. Weanling male Sprague-Dawley rats were randomly divided into eight dietary groups according to the type of dietary fat (beef tallow or corn oil) and level of dietary copper (0.74 or 6.9 microg Cu/g diet) or iron (44.4 or 86.7 microg Fe/g diet). Beef tallow and copper deficiency alone increased hepatic iron levels, which in turn were associated with increased plasma cholesterol. When the three dietary factors were combined, ie, iron, beef tallow, and copper deficiency, they induced the highest magnitude of hepatic iron retention, which in turn was associated with the highest concentration of plasma cholesterol. In contrast, when hepatic iron retention was not increased, such as by feeding a diet containing corn oil or by consumption of a copper-adequate diet, plasma cholesterol was not elevated. Based on these data, it is suggested that nutrients that have the ability to increase hepatic iron have the potential to increase plasma cholesterol.

Beta-carotene 15,15'-dioxygenase activity is responsive to copper and iron concentrations in rat small intestine.

OBJECTIVE: Previous in vitro studies have suggested that beta-carotene 15,15'-dioxygenase is an iron-dependent enzyme. However, in vivo, it is difficult to alter iron tissue concentration by varying dietary iron because of homeostatic control. On the other hand, an interaction between iron and copper has been shown, i.e., copper-deficiency results in an increase of iron in rat liver. Therefore, we hypothesized that intestinal iron concentration could be increased by copper-deficiency. Our objective was to examine the effects of iron as affected by dietary copper on beta-carotene 15,15'-dioxygenase activity in the small intestine. METHODS: Weanling male Sprague-Dawley rats (40 to 45g) were divided into four dietary groups: two copper-adequate groups (6.0 microg Cu/g diet) and two copper-deficient groups (0.6 microg Cu/g) combined with either normal iron (44 microg Fe/g) or high iron (87 microg Fe/g). Iron and copper concentrations were determined by atomic absorption spectrophotometry and the dioxygenase activity by reverse phase HPLC. RESULTS: Intestinal copper concentration was significantly reduced (40%) by the consumption of the copper-deficient diets, but intestinal iron was not changed by doubling dietary iron in rats fed either copper-adequate or copper-deficient diets. However, as hypothesized, the two copper-deficient groups exhibited higher intestinal iron concentration (> or =137%, p<0.001) than the copper-adequate controls. In addition, intestinal beta-carotene 15,15'-dioxygenase activity was increased by 27% and 106%, respectively, for copper-deficient rats fed either normal or high iron diets, compared to the respective copper-adequate controls (p<0.01). The dioxygenase activity was not significantly affected by dietary iron in either copper-adequate or copper-deficient groups. Finally, the enzyme activity was positively correlated (r=0.67, p<0.0001) with iron concentration and negatively correlated (r=-0.49, p<0.01) with copper concentration in small intestine. CONCLUSIONS: Intestinal beta-carotene 15,15'-dioxygenase may be an iron-dependent enzyme sensitive to copper status in vivo.

Level of dietary iron, not type of dietary fat, is hyperlipidemic in copper-deficient rats.

OBJECTIVE: This study was conducted to determine whether high dietary iron will negate the protective effect of unsaturated fat against hyperlipidemia. METHODS: Forty-eight weanling, male Sprague Dawley rats were randomly assigned to eight dietary groups differing in the levels of copper and iron and type of dietary fat (saturated or unsaturated). The diets were either deficient (0.6 microg Cu/g) or adequate (6.8 microg Cu/g) copper and either adequate (53 microg Fe/g) or high (506 microg Fe/g) iron. All diets contained starch as the sole source of dietary carbohydrate. RESULTS: Regardless of the type of dietary fat, three copper-deficient rats fed the high levels of dietary iron died prematurely due to ruptured hearts. Surviving rats belonging to the copper deficiency and high-dietary iron regimen developed severe anemia, enlarged hearts and livers, and exhibited the highest levels of liver iron. These rats also developed hypercholesterolemia. Triglycerides were elevated by the consumption of high iron diets. CONCLUSION: Data show that levels of dietary iron, not the type of dietary fat, are potential inducers of hypertriglyceridemia. Data also show that the combination of high iron intake and dietary copper deficiency is responsible for elevating blood cholesterol.

Functional outcome of arthrodesis for failed total knee arthroplasty.

To establish the ability of a salvage procedure to restore an independent lifestyle, the SF-36 functional outcome instrument and the pain, mobility, and physical activity subscales of the Arthritis Impact Measurement Scale (AIMS) was used to assess patient function. Nine patients (10 knees) who had undergone arthrodesis for failed total knee arthroplasty were compared with a control group of successful primary total knee arthroplasty patients. Average clinical follow-up was 42 months (minimum: 24 months). For six of the eight SF-36 categories, the average scores for the arthrodesis and arthroplasty groups were similar. The average global scores for the two groups were nearly identical. The arthroplasty patients scored better on the AIMS physical activity and mobility subscales than the arthrodesis group, although the latter group fared better on the pain subscale. Overall, global scores favored the arthroplasty patients. The only subscales to show a statistically significant difference between the arthrodesis and arthroplasty groups were the SF-36 physical functioning and the AIMS physical activity subscales. This pilot study demonstrated the ability of a salvage procedure to allow for an independent lifestyle with minimal complications. Furthermore, despite its popularity, the SF-36 does not appear as sensitive as the AIMS to differences in functional status or health outcomes between total knee arthroplasty and arthrodesis patients.

Effect of hepatic iron on hypercholesterolemia and hypertriacylglycerolemia in copper-deficient fructose-fed rats.

The purpose of this investigation was to establish whether plasma cholesterol and triacylglycerol(s) in copper deficiency can be increased or decreased by hepatic iron levels. Weanling male Sprague-Dawley rats were randomly divided into six dietary groups based on levels of dietary copper and iron. They were fed from weaning their respective diets for 6 wk. Forty percent of the copper-deficient rats fed a 15.7 mumol Fe/g diet died; 22% of those fed a diet containing 8.6 mumol Fe/g died; and there were no deaths in the 3.4 mumol Fe/g diet group. Rats belonging to the group fed the high-iron diet also exhibited the highest levels of liver iron, liver glutathione, and plasma cholesterol and triacylglycerol(s) compared with those fed either the adequate or low levels of dietary iron. There was a direct correlation (r = 0.82 and 0.77, respectively) between levels of cholesterol and triacylglycerol(s) in plasma and hepatic iron concentrations. These results provide strong evidence that points to a major involvement of iron in the lipemia of copper deficiency. These data may be important to those individuals who consume large quantities of fortified iron foods and supplement with iron but whose intake of copper is suboptimal.

Benfluorex, a hypotriglyceridemic drug, reduces lipid peroxidation and alleviates adverse metabolic complications of copper deficiency.

The pathologies associated with copper deficiency in rats fed fructose may be induced, in part, by hypertriglyceridemia and lipid peroxidation. Reducing triacylglycerol levels in plasma may result in lowering lipid peroxidation, which in turn could ameliorate metabolic effects resulting from the combination of fructose feeding and copper deficiency. Benfluorex, a hypolipidemic factor able to reduce hypertriglyceridemia, was administered to weanling male rats fed either copper-deficient (0.6 microgram Cu/g) or adequate (6.0 micrograms Cu/g) diets containing fructose as the sole dietary carbohydrate. In copper-deficient rats, benfluorex (50 micrograms.kg-1.d-1) reduced plasma triacylglycerols from 45 to 31 mg/dL, reduced lipid peroxidation by approximately 50%, and prevented the enlargements of heart and liver size and the atrophy of the pancreas, and ameliorated anemia. It is suggested that lipid peroxidation associated with hypertriglyceridemia may be responsible for the pathologies induced by the combination of fructose consumption and copper deficiency.

Characterization of the rpoC gene of Streptomyces coelicolor A3(2) and its use to develop a simple and rapid method for the purification of RNA polymerase.

The Streptomyces coelicolor rpoC gene, that encodes the beta' subunit of RNA polymerase, was isolated using the Escherichia coli rpoC gene as a hybridization probe. Comparison of the predicted amino acid sequence of the S. coelicolor beta' subunit to those characterized from other bacteria revealed three distinct subfamilies of beta' subunits, one of which consists of the S. coelicolor subunit and those from Mycobacterium leprae and Mycoplasma genitalium. Using site-directed mutagenesis, the carboxy terminus of the S. coelicolor beta' subunit was modified to contain six histidine residues. The histidine-tagged gene, rpoCHIS, was used to replace the wild-type allele in the chromosome of S. coelicolor and S. lividans. These strains were unaffected in growth and sporulation, demonstrating that the histidine-tagged RNA polymerase was competent to carry out all essential in-vivo functions. During a 1-day procedure, highly purified RNA polymerase was obtained by nickel-NTA agarose affinity chromatography followed by heparin-sepharose chromatography. Using in-vitro run-off transcription assays, the affinity purified RNA polymerase was shown to initiate transcription correctly from the S. lividans galP1 and galP2 promoters, and the Bacillus subtilus veg and ctc promoters. An extension of this procedure yielded highly-purified core RNA polymerase. To facilitate introduction of the rpoCHIS allele into other genetic backgrounds, a mutation in the adjacent gene, rpoB (rifA), conferring rifampin-resistance, was isolated in S. coelicolor to provide a genetic marker to follow transfer of the rpoCHIS allele. The use of this affinity chromatography procedure, in combination with the ability to introduce the rpoCHIS allele into different Streptomyces strains by transformation, will greatly facilitate the in-vitro analysis of transcription in members of this genus.

Impaired endocrine and exocrine pancreatic functions in copper-deficient rats: the effect of gender.

OBJECTIVE: To examine the effect of gender on endocrine and exocrine pancreatic function in female and male rats fed from weaning a copper-deficient diet. METHODS: Weanling male and female rats were fed a copper-deficient or adequate diet for 4 weeks. Rats were sacrificed after an overnight fast. Livers and pancreata were removed, weighed and the concentrations of copper and iron were determined. In addition, insulin was measured in pancreatic tissue and plasma. Lipase and amylase activities were measured in pancreas. Lipid peroxidation was assessed in liver. RESULTS: Copper deficiency in the male resulted in a profound reduced glandular mass of the pancreas. The pancreas continued low activities of lipase and amylase but excessive levels of insulin. Iron retention in the pancreas of the copper-deficient male rat was greater than in the female counterpart. Effects of copper deficiency in female rats on pancreas mass and endocrine pancreas were of lesser magnitude compared with males. Plasma insulin in the female rat was much higher than in the male rat. Hepatic lipid peroxidation was increased by copper deficiency in the male rat but was unaffected in the female. CONCLUSIONS: Data show that pancreatic atrophy is more pronounced in males compared with females, and the endocrine pancreas of the male is more susceptible to dietary copper deprivation than the female rat. The greater degree of pancreatic atrophy and associated abnormalities in males compared with females may be related to the greater retention of pancreatic iron and subsequent peroxidative damage.

Effects of grafting on porous metal ingrowth. A canine model.

Twenty-five mongrel dogs were studied using implantation of autograft, fresh-frozen allograft, and beta-tricalcium phosphate around a porous-coated chrome-cobalt plug in the distal femoral metaphysis; interference-fit and overreamed control specimens were also studied. Over the course of this 4-month study, bone ingrowth through the grouting materials into the center plug was noted for autologous, allograft, and ceramic specimens. Quantitatively, in terms of push-out strength and histology, there were no significant differences between grafted groups; significantly higher push-out strengths were attained in each grafted subgroup compared with nongrafted, overreamed control subjects. In the setting of uncemented revision total hip arthroplasty, bone-grafting is frequently required. Because of the limited availability of autogenous bone and the potential liabilities of allograft material, attention has been given to bone substitutes. On the basis of this preliminary study, bone ingrowth into a porous metal substrate has been documented to occur through autograft, allograft, or ceramic grouting agents. Within the limits of this nonloaded experimental model, it appears that these materials are comparable in terms of their osteoconductive capability. Even in the optimal laboratory situation, bone ingrowth does not appear to occur in a canine model across a nongrafted 2-mm gap with regularity over a 16-week period.

Hepatic iron overload may contribute to hypertriglyceridemia and hypercholesterolemia in copper-deficient rats.

The present study was conducted in order to determine whether hepatic iron retention in rats fed a copper-deficient diet containing fructose is associated with hypertriglyceridemia and hypercholesterolemia, and whether a reduction of iron intake will prevent elevation of blood triglycerides and cholesterol. Rats were fed from weaning either a copper-deficient (0.6 microgram Cu/g) or copper-adequate (6.0 micrograms Cu/g) diet for 4 weeks. Half the rats consumed either an adequate level of iron (50 micrograms Fe/g) or a low level (17 micrograms Fe/g). Reduction of iron intake reduced blood levels of both triglycerides and cholesterol in rats fed a copper-deficient diet containing fructose. In addition, hepatic lipid peroxidation was also decreased. The combination of high iron, low copper, and fructose may be responsible for increased levels of risk-factor metabolites associated with heart disease.

Metal carcinogenesis in total joint arthroplasty. Animal models.

As early as 1956, laboratory investigations into the carcinogenicity of modern dental and orthopaedic alloys were undertaken. Such studies were prompted by the observation that workers, particularly in nickel and chromate refining, had increased risks of nasal and lung tumors. For the past 25 years, sporadic case reports have documented the development of malignant neoplasms proximate to an orthopaedic implant. Although the results of epidemiologic studies have not shown an excessive number of tumors in patients receiving stainless steel or superalloy implants, the possibility of carcinogenesis, given the corrosive environment in which metal implants exist, has prompted ongoing laboratory studies. Leaching of metal ions from implants, the synovial processing of metallic wear debris, and the effects of exposure to intraarticular metal particles have been the subjects of numerous laboratory studies. The results of these studies are summarized and recommended parameters for future laboratory investigations are given.

Oerskovia xanthineolytica infection of a prosthetic joint: case report and review.

Oerskovia spp. are gram-positive, Nocardia-like bacilli which inhabit the soil and rarely cause human infections. Previously reported cases of Oerskovia infection have been characterized by a nonaggressive course and an association with foreign bodies. We report the first case of a patient with a prosthetic joint infection due to Oerskovia xanthineolytica. Our patient presented with a prolonged, indolent course and was thought to have aseptic loosening of his prosthesis until the time of surgery. He was cured of his infection by removal of the prosthesis, antibiotic therapy, and delayed reimplantation. Review of the previous 10 reported cases of Oerskovia infection in humans supports the recommendation that foreign-body-associated infections should be treated with a strategy that includes removal of the foreign material.

Responses of insulin to oral glucose and fructose loads in marginally copper-deficient rats fed starch or fructose.

The purpose of this study was to assess the effects of dietary fructose either alone or in combination with marginal copper deficiency in weanling male rats exposed to their respective diets for only 2 wk. This short duration of exposure to inadequate copper intake prevents progressive morbidity brought about by increasing periods of exposure to dietary copper deprivation. Weanling male rats were fed a copper-deficient (0.6 microgram Cu/g) or a copper-adequate (6.0 micrograms Cu/g) diet containing 62% fructose or 62% starch for 2 wk. Either an oral glucose or an oral fructose tolerance test was conducted after an overnight fast. Insulin levels were elevated by either oral glucose or oral fructose at fasting and at 30 min postload in rats fed fructose compared with those fed starch. Despite high levels of plasma, insulin blood glucose was not reduced. Marginal copper deficiency had no effect on either plasma insulin or blood glucose. Data identify fructose as the sole agent responsible for inducing adverse changes in glucose metabolism. Two weeks of fructose consumption was sufficient to produce these changes.

Antioxidant defense system in lung of male and female rats: interactions with alcohol, copper, and type of dietary carbohydrate.

Male and female rats were used to investigate the effects of type of dietary carbohydrate (CHO), copper, and ethanol consumption on lung antioxidant enzyme activities and levels of phosphorylated compounds in whole blood. Copper-deficient female rats exhibited a greater degree of copper deficiency than males, as assessed by hepatic copper concentration and hepatic copper superoxide dismutase (CuSOD) activity. However, copper-deficient male rats fed fructose-containing diets exhibited greater growth retardation, anemia, and heart hypertrophy than females consuming the same diets and males fed starch. In addition, one of 10 copper-deficient male rats that ate a fructose-based diet and drank water and one of 10 copper-deficient male rats that ate a starch-based diet and drank ethanol died. Copper-deficient, starch-fed males exhibited the highest activities of glutathione peroxidase (GSH-Px) and catalase as compared with fructose-fed rats. Ethanol consumption elevated the activities of GSH-Px and catalase. Copper-deficient female rats exhibited higher catalase but lower GSH-Px activities than males. It is suggested that in copper deficiency, the ability to increase antioxidant enzyme activities in rats consuming starch is greater than in rats consuming fructose. Rats fed starch are provided with a greater degree of protection against oxidative damage than rats fed fructose. In addition, polyphosphorylated compounds in blood were reduced in copper-deficient male rats that consumed fructose-based diets. This may impair supply of oxygen to tissues.