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Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVIII | Pages 162 - 162
1 Sep 2012
Bernstein M Desy NM Huk OL Zukor DJ Petit A Antoniou J
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Purpose

Metal-on-metal (MoM) articulations in total hip replacement (THR) have become an attractive option for young, active patients. Short-term reports have demonstrated elevated systemic metal ion levels in the blood and urine. Disseminated concentrations of cobalt and chromium have raised concern regarding cellular toxicity, chromosomal damage and adverse local soft tissue reactions.

Long-term studies are required to support the increased use of MoM bearings in younger patients given their potential deleterious effects. The purpose of the current study was to report the seven to 13 year clinical, radiographic, and metal ion results in patients following MoM THR.

Method

We prospectively followed 165 patients (196 hips) after second-generation MoM THR between July 1997 and November 2003. Functional outcome was measured using the Harris Hip Score (HHS) and the University of California Los Angeles (UCLA) Activity Score. Radiographic analysis was performed using Einzel-Bild-Roentgen-Analyse (EBRA) by two of the authors blinded to the study. Cobalt and chromium metal ions were measured from whole blood and analyzed using inductively coupled plasma-mass spectrometry as previously described.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVIII | Pages 165 - 165
1 Sep 2012
Powell JN Beaulé PE Antoniou J Bourne RB Schemitsch EH Vendittoli P Smith F Werle J Lavoie G Burnell C Belzile É Kim P Lavigne M Huk OL O'Connor G Smit A
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Purpose

The purpose of the study was to determine the rate of conversion from RSA to THR in a number of Canadian centers performing resurfacings

Method

Retrospective review was undertaken in 12 Canadian Centers to determine the rate of revision and reason for conversion from RSA to THR. Averages and cross-tabulation with Chi-Squared analysis was performed. kaplan Meier survivorship was calculated.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVII | Pages 341 - 341
1 Sep 2012
Bernstein M Desy N Huk O Zukor D Petit A Antoniou J
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Introduction

Metal-on-metal (MoM) articulations in total hip replacement (THR) have become an attractive option for young, active patients. Short-term reports have demonstrated elevated systemic metal ion levels in the blood and urine. Disseminated concentrations of cobalt and chromium have raised concern regarding cellular toxicity, chromosomal damage and adverse local soft tissue reactions.

Long-term studies are required to support the increased use of MoM bearings in younger patients given their potential deleterious effects. The purpose of the current study was to report the 7–13 year clinical, radiographic, and metal ion results in patients following MoM THR.

Methods

We prospectively followed 165 patients (196 hips) after second-generation MoM THR between July 1997 and November 2003. Functional outcome was measured using the Harris Hip Score (HHS) and the University of California Los Angeles (UCLA) Activity Score. Radiographic analysis was performed using Einzel-Bild-Roentgen-Analyse (EBRA) by two of the authors blinded to the study. Cobalt and chromium metal ions were measured from whole blood and analyzed using inductively coupled plasma-mass spectrometry.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVIII | Pages 60 - 60
1 Sep 2012
Rampersad S Petit A Ruiz JC Wertheimer MR Antoniou J Mwale F
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Purpose

A major drawback of current cartilage and intervertebral disc (IVD) tissue engineering is that human mesenchymal stem cells (MSCs) from osteoarthritic (OA) patients express high levels of type X collagen. Type X collagen is a marker of late stage chondrocyte hypertrophy, linked with endochondral ossification, which precedes bone formation. However, it has been shown that a novel plasma-polymer, called nitrogen-rich plasma-polymerized ethylene (PPE:N), is able to inhibit type X collagen expression in committed MSCs. The aim of this study was to determine if the decreased expression of type X collagen, induced by the PPE:N surfaces is maintained when MSCs are removed from the surface and transferred to pellet cultures in the presence of serum and growth factor free chondrogenic media.

Method

Human MSCs were obtained from aspirates from the intramedullary canal of donors undergoing total hip replacement for OA. Cells were expanded for 2–3 passages and then cultured on polystyrene dishes and on two different PPE:N surfaces: high (H) and low (L) pressure deposition. Cells were transferred for 7 additional days in chondrogenic serum free media (DMEM high glucose supplemented with 2 mM L-glutamine, 20 mM HEPES, 45 mM NaHCO3, 100 U/ml penicillin, 100 ug/ml streptomycin, 1 mg/ml bovine serum albumin, 5 ug/ml insulin, 50 ug/ml ascorbic acid, 5 ng/ml sodium selenite, 5 ug/ml transferrin) in pellet culture or on PS cell culture dishes. RNA was extracted using a standard TRIzol protocol. RT-PCR was realized using Superscript II (RT) and Taq polymerase (PCR) with primers specific for type I and X collagen. GAPDH was used as a housekeeping gene and served to normalize the results.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVIII | Pages 34 - 34
1 Sep 2012
Gawri R Mwale F Ouellet J Steffen T Roughley P Haglund L Antoniou J
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Purpose

Disc degeneration is known to occur early in adult life, but at present there is no medical treatment to reverse or even retard the problem. Development of medical treatments is complicated by the lack of a validated long term organ culture model in which therapeutic candidates can be studied. The objective of this study was to optimize and validate an organ culture system for intact human intervertebral disc (IVD), which could be used subsequently to determine whether synthetic peptide growth factors can stimulate disc cell metabolism and initiate a repair response.

Method

Seventy lumbar IVDs, from 14 individuals, were isolated within 24 h after death. Discs were prepared for organ culture by removing bony endplates but retaining cartilaginous endplates (CEP). Discs were cultured with no external load applied. The effects of glucose and FBS concentrations were evaluated. Dulbeccos Modified Eagle Media (DMEM) was supplemented with glucose, 4.5g/L or 1g/L, referred to as high and low (physiological) glucose, and FBS, 5% or 1%, referred to as high and low FBS, respectively. After a four week culture period, samples were taken across the disc using a 4 mm biopsy punch. Cell viability was analyzed using a live/dead fluorescence assay (Live/Dead, Invitrogen) and visualized by confocal microscopy. CEP discs were also placed in long term culture for four months, and cell viability was assessed. Western bolt analysis for the G1 domain of aggrecan was also performed to assess the effect of nutritional state on disc catabolism.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVIII | Pages 67 - 67
1 Sep 2012
Mwale F Petit A Yao G Antoniou J
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Purpose

Whilst it is known that oxidative stress can cause early degenerative changes observed in experimental osteoarthritis and that a major drawback of current cartilage and intervertebral disc tissue engineering is that human mesenchymal stem cells (MSCs) from osteoarthritis (OA) patients express type X collagen, a marker of late-stage chondrocyte hypertrophy (associated with endochondral ossification), little is known whether the expression of type X collagen in MSCs from OA patients can be related to oxidative stress or inflammatory reactions that occur during this disease.

Method

Human MSCs were obtained from aspirates from the intramedullary canal of donors undergoing total hip replacement for OA. Bone marrow aspirates were processed essentially as previously described. Briefly, non-adherent cells were discarded after 72h of culture and the adherent ones were expanded for 2–3 passages. MSCs from normal donor (control) were obtained from Lonza. Cells were then lysed and protein expression was detected by Western blot using specific antibodies directed against type X collagen, as well as the antioxidant enzymes Mn-superoxide dismutase (MnSOD), catalase (CAT) and glutathione peroxidase-1 (GPx-1) and inflammation related proteins cyclooxygenase-1 (COX-1) and intercellular adhesion molecule-1 (ICAM-1). GAPDH was used as a housekeeping gene and served to normalize the results. Correlations between the expressions of the different proteins were realized using the correlation Z test with StatView (SAS Institute).


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_IV | Pages 567 - 567
1 Nov 2011
Beaulé PE Smith F Powell JN Antoniou J Bourne RB Lavigne M Belzile E Schemitsch EH Garbuz D
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Purpose: Recently, there has been concern raised on the occurrence of pseudotumors after metal on metal hip resurfacing. A pseudotumor is defined as a local soft tissue mass associated with localized bony and/or tissue (muscle) destruction. The primary purpose of this study is to determine the incidence of this complication in several high volume Canadian academic centres.

Method: Nine of the 11 Canadian academic centres who perform metal on metal hip resurfacings were surveyed. The number of metal on metal hip resurfacing arthroplasties performed at each centre was first determined, as were the number of those who have presented with a pseudotumour, and subsequently gone on to revision surgery. The basic demographics of the group were recorded, as were the radiographic and implant design variables for those cases presenting with a pseudotumour.

Results: A sample of 3,400 hip resurfacing arthroplasties performed between 2002 and December 2008 were surveyed. Demographics were tabulated for a sub-sample of these patients. 76% were male, the mean length of follow-up was 3.02 years, mean BMI was 28.65, and mean age was 52.10 years. Three of 3,400 cases presented with a pseudotumour, an incidence of .09%.

Conclusion: Although pseudotumors remain a concern after metal on metal hip resurfacing, the incidence at short to mid term follow-up is very low in this multi-centre academic survey. This information is significantly lower than what other groups have recently reported. Continued close monitoring is required in order to determine what clinical factors are at play.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_IV | Pages 567 - 567
1 Nov 2011
Antoniou J Petit A Nikolaou VS Papanastasiou C Mwale F Zukor DJ Huk OL
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Purpose: Several studies have shown elevated levels of metal ions in blood of patients with metal-on-metal (MM) total hip arthroplasty (THA). The outstanding question that remains is the clinical impact of these elevated ion levels. Even though it is well known that exposure to heavy metals such as lead, copper, mercury, nickel, and cadmium) may lead to significant alterations in human sperm morphology and motility, less is known on the effect of Co and Cr on semen parameters. The aim of the present study was to investigate the effect of metal ions on the semen of males of child fathering age with MM hip arthroplasty.

Method: Semen was collected form 10 patients between 41 and 49 years old (mean = 45±6 years) by masturbation after 2–3 days of abstinence. Samples were examined within 1h after ejaculation for morphology, motility, and number of sperm cells following standard criteria from the World Health Organization (WHO). Co and Cr concentrations were measured in both the seminal plasma and in the blood of patients by inductively coupled plasma-mass spectroscopy (ICP-MS). Since spermatozoa membrane polyunsaturated fatty acids are vulnerable to attack by reactive oxygen species (leading to peroxide formation), peroxide concentrations were measured in both the seminal plasma and the blood of patients.

Results: Results showed that the concentration of both Co and Cr ions was significantly lower in the seminal plasma than in the blood of the patients. Results also showed that the levels of peroxides were lower in the seminal plasma than in the blood plasma of these patients. Importantly, the ejaculate volume, the sperm density, the total sperm count, the pH, and the percentage of cells with normal morphology were in the range of the WHO criteria for fertile population and also in the range of reference patients in the city of measurements. However, the viability was a little bit lower than what was observed in a fertile population without prosthesis.

Conclusion: The presence of Co and Cr ions in the blood of males of child fathering age with MM hip arthroplasty raised concerns about the quality of semen in these patients. Results of the present study strongly suggest that the raised of Co and Cr had no significant effect on sperm parameters of young patients with MM prosthesis. The methods used to identify potential normal and fertile semen samples are still contradictory and not exactly defined. Studies showed for example that only total numbers of sperm with progressive mobility are significantly different in the fertile than in sub-fertile men, while others suggested that the fertile population should be defined by sperm concentration or sperm morphology. In conclusion, results suggest that Co and Cr ions generated from MM prosthesis have no significant effect on the sperm parameters of young patients of child fathering age. Further longitudinal studies are however necessary to conclusively determine the effect of metal ions from MM prosthesis on sperm parameters.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_IV | Pages 563 - 564
1 Nov 2011
Rampersad S Petit A Yao G St-Georges-Robillard A Ruiz J Wertheimer MR Antoniou J Mwale F
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Purpose: Several studies have been directed toward using mesenchymal stem cells (MSCs) from osteoarthritic (OA) patients for cartilage or disc repair because these patients are the ones that will require a source of autologous stem cells if biological repair of tissue lesions is to be a therapeutic option. A major drawback of current cartilage and intervertebral disc tissue engineering repair is that these cells rapidly express type X collagen, a marker of late stage chondrocyte hyperthrophy implicated in endochondral ossification. However, a novel plasma-polymerized thin film material, named nitrogen-rich plasma-polymerized ethylene (PPE:N), is able to inhibit type X collagen expression in committed MSCs. The specific aim of this study was to determine if the suppression of type X collagen by PPE:N is maintained when MSCs are transferred to pellet cultures in chondrogenic defined media.

Method: MSCs were obtained from aspirates from the intramedullary canal of donors undergoing total hip replacement for OA using a protocol approved by the Research Ethics Committee of our institution. Cells were then expanded for 2–3 passages in DMEM high glucose supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin, and finally cultured on polystyrene (PS) cell culture dishes or PPE: N surfaces for 3 and 7 days. Cells were transferred for 3 additional days in a chondrogenic serum free media (DMEM high glucose supplemented with 2 mM L-glutamine, 20 mM HEPES, 45 mM NaHCO3, 100 U/ml penicillin, 100 μg/ml streptomycin, 1 mg/ml bovine serum albumin, 5 μg/ml insulin, 50 μg/ml ascorbic acid, 5 ng/ ml sodium selenite, 5 μg/ml transferrin) in pellet culture or on PS cell culture dishes. Cells were then lysed and proteins were separated on 4–20% acrylamide gels and transferred to nitrocellulose membranes. Type X collagen was detected by Western blot; GAPDH expression was used as an internal control for protein loading.

Results: Results showed that type X collagen protein was expressed in MSCs from OA patients cultured on polystyrene but was suppressed when cultured on PPE: N. Since defined chondrogenic medium are commonly used in pellet culture to promote in vitro chondrogenesis, we then investigated the effect of transferring cells pre-cultured on PPE:N into pellet culture on type X collagen expression. However, the decreased type X collagen expression was not maintained in these conditions and that the expression returned to control values. The decreased type X collagen expression was maintained when the cells were cultured on PS cell culture dishes.

Conclusion: The use of MSCs is promising for tissue engineering of cartilage and intervertebral disc. The present study confirmed the potential of PPE:N surfaces in suppressing type X collagen expression in MSCs from OA patients. However, when MSCs stem cells are transferred to pellet cultures, type X collagen is rapidly re-expressed suggesting that pellet cultures may not be suitable for chondrogenesis of MSCs from OA patients.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_IV | Pages 562 - 562
1 Nov 2011
Mwale F Yao G Petit A Antoniou J
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Purpose: Mesenchymal stem cells (MSCs) from osteoarthritic (OA) patients are not well characterized and little is known of how they are regulated. Recent evidence indicates that a major drawback of current cartilage and intervertebral disc (IVD) tissue engineering is that human MSCs from OA patients express type X collagen (COL10), a marker of late-stage chondrocyte hypertrophy (associated with endochondral ossification). However, the intracellular pathways for transducing signals that regulate hypertrophy in MSCs remain unclear. In chondrocytes, this pathway is mediated by mitogen activated protein kinase (MAPK) p38. The aim of this study was to determine the phosphorylation levels of ERK/p38 MAPK signaling molecules in MSCs from OA patients compared to those from normal patients.

Method: MSCs were obtained from aspirates from the intramedullary canal of donors (60–80 years of age) undergoing total hip replacement for OA. Cells were cultured in DMEM high glucose supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin for 2–3 passages. Cells were then lysed and proteins were separated on 10% acrylamide gels and transferred to nitrocellulose membranes. Protein expression was determined by Western blot using specific antibodies directed against type X collagen, ERK, phosphorylated-ERK, p38, phosphorylated-p38, JNK, phosphorylated-JNK, AKT, and phosphorylated-AKT. GAPDH was used as a housekeeping gene. Proteins were detected using the West Pico Chemiluminescence substrates and analyzed using the Bio-Rad VersaDoc equipped with a cooled CCD 12 bit camera. Normal mesenchymal stem cells from a 22 years old woman were purchased from Lonza (Switzerland).

Results: Results show that the expression of COL10 was markedly increased in MSCs of OA patients compared to control patient. Results also shows that the phosphorylation of all the signal transduction proteins studied was induced in MSCs of patients with OA. Indeed, the phosphorylation of ERK (3.4±0.9 times the control), p38 (1.7±0.3 times the control), JNK (5.40±1.14 times the control), and AKT (4.3±0.8 times the control) was higher in MSCs of OA patients compared to control normal patients.

Conclusion: In the normal donor, MSCs continue to exhibit their in situ behavior in that they expressed very little or no COL10. This may relate to the fact that normal MSCs being multipotent in nature like to maintain an undifferentiated state. In contrast, MSCs from OA patients expressed COL10: this suggests that they are in a situation were they can be preprogrammed not only to replace the degraded articular cartilage but also the damaged subchondral bone. Since the phosphorylation of ERK/p38 MAPK signaling molecules is also lower in normal MSCs, our results also suggest that this signaling pathway is implicated in the control of COL10 expression. This finding is of great importance for the understanding of COL10 regulation in general and may lead to important advances in the comprehension of COL10 related diseases.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_IV | Pages 564 - 564
1 Nov 2011
Mwale F Petit A Yao G Antoniou J
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Purpose: A major drawback of current cartilage and intervertebral disc tissue engineering is that human mesenchymal stem cells (MSCs) from osteoarthritis (OA) patients express type X collagen (COL10), a marker of late-stage chondrocyte hypertrophy (associated with endochondral ossification). Parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHrP) regulate endochondral ossification by inhibiting chondrocyte differentiation toward hypertrophy. In the present study, we investigated the effect of PTH on the expression of COL10 in MSCs from OA patients and analyzed the potential mechanisms related to its effect.

Method: MSCs were obtained from aspirates from the intramedullary canal of donors (60–80 years of age) undergoing total hip replacement for OA. Cells were cultured for 2–3 passages in DMEM high glucose supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin. Cells were then incubated for 0–24h without (Control) or with 100 nM PTH (1–34). Cells were lysed and proteins were separated on 10% acrylamide gels and transferred to nitrocellulose membranes. Protein expression was detected by Western blot using specific antibodies directed against COL10, p38, phosphorylated-p38 (p-p38), SAP/JNK, phosphorylated-SAP/JNK (p-JUNK). GAPDH was used as a housekeeping gene. Protein levels were analyzed using a Bio-Rad VersaDoc equipped with a cooled CCD 12 bit camera.

Results: Results showed that PTH inhibited in a time-dependent manner the expression of COL10 in MSCs from OA patients. The level of expression reached 21% of control (79% inhibition) after 24h. This inhibitory effect of PTH was reversed by Calphostin C, an inhibitor of protein kinase C. To further investigate the mechanism of action related to the effect of PTH on COL10 expression, we measured the phosphorylation of p38 and showed that PTH also inhibited this phosphorylation, which is an indicator of its activity. The level of phosphorylation reached 74% of control after 3h and stayed stable thereafter. Similarly, treatment of MSCs with PTH suppressed the phosphorylation of JNK, another major stress-activated MAP kinase. The level of phosphorylation reached 65% of control after 6h and returned to control values after 24h.

Conclusion: Results of the present study suggested that PTH may be a potential regulator of COL10 expression in MSCs from OA patients. Results also suggested a role for the protein kinase C and the p38/JNK pathways in this regulation. p38 and JNK are serine and threonine protein kinases that are activated by osmotic pressure, stress, and cytokines. It is therefore not surprising that their activities were elevated as OA (degenerative joint disease) is a result of trauma or infection to the joint and is characterized by an up-regulation of cytokines. Further studies are however necessary to better understand the role of these molecules in hypertrophy.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_IV | Pages 575 - 576
1 Nov 2011
Antoniou J Petit A Mwale F Zukor DJ Huk OL
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Purpose: Several studies have shown elevated levels of metal ions in blood of patients with metal-on-metal (MM) total hip arthroplasty (THA). To minimize wear, the elastohydrodynamic theory suggests wear reduction for larger diameter head bearings. This significant reduction in wear has been demonstrated in hip simulators for the 36 mm-head compared to the 28 mm-head prosthesis. However, the survival of larger head MM THAs and the levels of metal ions in the blood of patients having these implants are still to be determined.

Method: Fifty (50) patients (56 hips) who received a DePuy Ultamet™ MM hip bearing (40/44 mm-head) at our Institution between July 1st 2007 and August 31st 2008 were included in the study. Clinical and radiologic data were collected pre-operatively as well as at 6–8 weeks, 4–6 months, and 1 year postoperatively. Results were compared to those of subjects (65 patients, 71 hips) who received a 36 mm-head prosthesis. Cobalt (Co) and chromium (Cr) concentrations were measured at 1 year post-operatively in the blood of patients by ICP-MS. Since Co and Cr ions have the potential to induce irreversible biochemical damage to macromolecules, the levels of oxidative stress markers (total antioxidants and lipid peroxides) were measured in the plasma of these patients.

Results: At their 1-year post-operatively follow-ups, all patients were doing well and no sign of osteolysis was observed on X-rays. Harris Hip Score increased in both groups with a tendency to higher score in the 40–44 mm group compared to the 36 mm group. Activity score also increased in both groups after 1 year without statistical significant differences. Results also show that the levels of Co and Cr ions increased significantly in both groups compared to the Pre-OP control group. The levels of Co were also significantly higher in patients with large head arthroplasty (40 and 44 mm-head) compared to those of the 36 mm-head group (p=0.012). The levels of Cr were similar in both the large head and the 36 mm-head group (p=0.41). Finally, results show that there were no differences in the levels of total antioxidants and peroxides between the 40–44 mm group and the 36 mm group. Moreover, there was no increase in the level of these markers of oxidative stress compared to the Pre-OP control group.

Conclusion: The present study shows that at 1-year postoperatively, patients with large 40–44 mm-head THA had comparable clinical outcomes than those with 36 mm-head prosthesis. However, the levels of Co ions were significantly higher in these patients compared to patients with 36 mm-head THA. This suggests a higher health risk for these patients due to the presence of these ions. However, there were no effects on the levels of oxidative stress markers in the blood of these patients, suggesting that there is no increased risk at short-term. In conclusion, due to the high level of Co ions, longer follow-ups are required to conclusively determine the outcomes of the patients and the survivorship of these new bearings.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_IV | Pages 563 - 563
1 Nov 2011
Mwale F Epure LM Yoshikawa T Hemmad A Bokar M Masuda K Roughley PJ Antoniou J
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Purpose: Intervertebral disc (IVD) degeneration is associated with proteolytic degradation of proteoglycan aggregates present within the extracellular matrix of the disc. Link-N peptide is the N-terminal peptide of link protein, which stabilizes the proteoglycan aggregates. It is generated in vivo by proteolytic degradation during tissue turnover. We have previously shown that this peptide can stimulate the synthesis of proteoglycans and collagens by IVD cells in vitro. However, to date, there have been no reports on the effect of Link-N on the IVD in vivo. The purpose of the present study was to determine the effect of intradiscally administration of Link-N peptide on disc cell survival and extracellular matrix synthesis using a rabbit annular needle puncture model of IVD degeneration.

Method: Twelve New Zealand white rabbits (~3.5 kg; 5–6 months old) received an annular puncture with an 18-gauge needle on 2 non-contiguous discs (L2–L3 and L4–L5). The disc (L3–L4) between the punctured discs and that above (L5–L6) was left intact as internal controls. Two weeks after the initial puncture, the anterior surfaces of the previously punctured discs (L2–L3 and L4–L5) were injected with either saline (10 μl/disc) or Link-N (100 μg in 10μl saline/disc) into the center of the NP. Disc height was radiographically monitored biweekly. After 12 weeks post-injection, all the rabbits were euthanized and the IVDs from both experimental groups were removed from each lumbar spine for biochemical analysis. The nucleus pulposus (NP) was separated from the annulus fibrosus (AF), the specimens weighed (wet weight), the content of DNA measured using PicoGreen, and the total contents of sulfated glycosaminoglycans (GAG) measured by the 1,9-dimethylmethylene blue (DMMB) assay.

Results: Following needle puncture that initiates disc degeneration, the disc height index (DHI) decreased by about 25%. By 6 weeks after Link-N injection, the mean percent DHI of injected discs in the Link-N group was higher than in the saline group. This difference in mean percent DHI was maintained during the rest of the follow-up. Puncturing the IVD also led to a decrease in proteoglycan content in both the NP and the AF in saline-treated discs. Treatment with Link-N stimulated proteoglycan synthesis (GAG) in both the NP and AF by about 20%. Link-N did not cause an increase in the DNA content of the discs.

Conclusion: Results of the present study show that Link-N can stimulate proteoglycan production in vivo when administered to degenerate disc. This stimulation occurs in both the NP and AF of the disc and in the absence of any effect on cell division. The changes observed with Link-N on proteoglycan synthesis are similar to those reported after injection of osteogenic protein-1 (OP-1) Thus, Link-N appears to be equally effective at stimulating repair of the IVD in vivo. One major advantage of Link-N over OP-1 for therapeutic use is the large saving in cost, Link-N being about 400 times cheaper than OP-1.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_III | Pages 247 - 247
1 Jul 2011
Antoniou J Mwale F Zukor DJ Huk OL Petit A
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Purpose: The presence of metal ions in the blood of patients with a metal-on-metal (MM) bearing points to the importance of understanding the long-term effects of these ions. Metal ions have the potential to induce the production of reactive oxygen species (ROS), making them prime suspects for inducing molecular damage in circulating cells. The aim of this study was to analyze the levels of oxidative stress markers in the plasma of patients with hip surface replacement.

Method: Blood was collected up to 3 years after implantation from 66 patients with articular surface replacement (ASRÔ, DePuy Orthopaedics) and 54 patients with 36 mm-head MM THA. Forty (40) pre-operative patients were also assessed as control group. Total anti-oxidant levels were measured by the Oxford Biomedical total antioxidant power assay (Oxford, MI) to obtain an overview of the defense capacity of patient’s oxidative stress. Peroxide concentrations were measured by the Biomedica OxyStat assay (Medicorp, Montreal, QC) to quantify damage to lipids in the systemic circulation. Nitrototyrosine levels were quantified using the BIOXYTECH® Nitrotyrosine-EIA assay (OxisResearch™, Portland OR) to measure damage to proteins. The concentrations of metal ions were analyzed by inductively coupled plasma-mass spectroscopy.

Results: Results showed that there were no statistical differences in the concentrations of total antioxidants, lipid peroxides, and protein nitrotyrosines between the control, the ASR, and the 36 mm-head groups. Furthermore, there was no correlation between the concentrations of these markers and the concentrations of both Co and Cr ions (r2 £ 0.006).

Conclusion: The single most significant obstacle preventing a broader application of metal-on-metal hip arthroplasties and resurfacings continues to be the concerns regarding elevated metal ion levels in the blood and urine of patients. The present results showed that there were no changes in the levels of oxidative stress markers in patients with MM bearings compared to the control group. Given the possible latency periods related to metal ion exposure, longer follow-ups are required to conclusively determine the effects of elevated circulating ions on oxidative stress in the blood of patients with MM bearings.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_III | Pages 244 - 244
1 Jul 2011
Mwale F Wang HT Girard-Lauriault P Wertheimer MR Antoniou J Petit A
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Purpose: Recent evidence indicates that a major drawback of current cartilage and intervertebral disc (IVD) tissue engineering is that human mesenchymal stem cells (MSCs) from osteoarthritic patients rapidly express type X collagen (COL10A1), a marker of late-stage chondrocyte hypertrophy associated with endochondral ossification. We recently discovered that a novel atmospheric-pressure plasma-polymerized thin film substrate, named “nitrogen-rich plasma-polymerized ethylene” (PPE:N), is able to inhibit COL10A1 expression in committed MSCs. However, the cellular mechanisms implicated in the inhibition of COL10A1 expression by PPE:N surfaces are unknown.

Method: Human mesenchymal stem cells (MSCs) were obtained from aspirates from the intramedullary canal of donors (60–80 years of age) undergoing total hip replacement for osteoarthritis. Bone marrow aspirates were processed and MSCs were cultured on commercial polystyrene (PS control) and on PPE:N surfaces in the presence of different kinases and cyclooxygenase inhibitors for 3 days. Total RNA was extracted with TRIzol reagent (Invitrogen, Burlington, ON) and the expression of COL10A1, cyclooxygenase-1 (COX-1), and 5-lipoxygenase (5-LOX) genes was measured by real-time quantitative RT-PCR.

Results: Results showed that a non-specific inhibitor of cyclooxygenases reduced the expression of COL10A1. In contrast, inhibitors of protein kinases stimulated the expression of COL10A1. Furthermore, potent and selective inhibitors of COX-1 and 5-LOX also reduced the expression of COL10A1. However, COX-2 and 12-LOX inhibitors had no significant effect on the expression of COL10A1. COX-1 gene expression was also decreased when MSCs were incubated on “S5” PPE:N surfaces. Interestingly, MSCs did not express 5-LOX.

Conclusion: PPE:N surfaces suppress COL10A1 expression through the inhibition of COX-1 which is directly implicated in the synthesis of prostaglandins. The decreased expression of COX-1 and COL10A1 in human MSCs cultured on PPE:N is therefore in agreement with the induction of the osteogenic capacity of rat bone marrow and bone formation by systemic or local injection of PGE2 in rats. However, PGE2 and other prostaglandins inhibited COL10A1 expression in chick growth plate chondrocytes. This suggests that the effect of prostaglandins on COL10A1 expression may be cell-specific or may be dependent on pre-existing patho-physiological conditions.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_II | Pages 130 - 130
1 May 2011
Nikolaou V Petit A Huk O Zukor D Bergeron S Antoniou J
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Introduction: Several studies have shown the presence of cobalt (Co) and chromium (Cr) ions in blood, urine, and organs of patients after THA using Co-Cr alloy-based implants. Even though it is well known that exposure to heavy metals may lead to significant alterations in human sperm morphology and motility, less is known on the effect of Co and Cr on semen parameters after metal on metal (MOM) hip replacement.

Methods: Semen was collected form 10 patients between 41 and 49 years old (mean=45.9±3.0 years) by masturbation after 2–3 days of abstinence. The time of implantation varied from 1 to 9 years (mean=5.1±3.9 years). Samples were collected in a sterile container and examined within 1h after ejaculation for morphology, motility, and number of sperm cells following standard criteria. All patients were doing well at their follow-up visits (Harris Hip Score=94±4; UCLA activity a score=7±1) and no sign of osteolysis was observed on X-rays.

Co and Cr concentrations were measured in both the seminal plasma and in the blood of patients by inductively coupled plasma-mass spectroscopy (ICP-MS).

Results: results showed that the levels of Co in the seminal plasma and the blood of the patients were not statistically different. However, the level of Cr was significantly lower in the seminal plasma than in the blood of the patients. The ejaculate volume (2.1 ±0.6 ml), the sperm density (66±53 x 106), the total sperm count (151±75 x 106/ml), the pH (8), and the percentage of normal morphology (46±18%) were in the range of the WHO criteria for fertile population and also in the range of reference patients in the city of measurements. However, the viability was lower than that observed in a fertile population without prosthesis (41±19%).

Conclusions: results of the present study strongly suggest that both Co and Cr ions crossover to the semen but that their concentrations were too low to significantly affect sperm parameters of young patients with MM prosthesis. Further longitudinal studies are however necessary to conclusively determine the effect of metal ions from MM prosthesis on sperm parameters.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_II | Pages 129 - 129
1 May 2011
Bernstein M Bergeron S Petit A Huk O Antoniou J
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Introduction: Metal on metal hip implants continue to be successful alternatives to conventional bearings in younger patients with osteoarthritis. Levels of metal ions such as cobalt (Co) and chromium (Cr) increase in patients with metal bearing hip replacements and resurfacings. These particles are cytotoxic, induce bone loss, and lead to malignant tumors in rats. A subset of these patients are considered outliers as they have unusually high levels of Co and Cr ions. Given the increasing prevalence of metal bearings and the potential for cellular toxicity, we attempted to determine whether patient or surgical factors could account for abnormally elevated ion levels.

Methods: We analyzed the Co and Cr levels from whole blood in 661 patients with metal on metal hip bearings. Patient outliers were defined as those who had ion levels ≥ three-fold the mean value. Twenty-four patients (3.6%) had abnormally high metal ion levels, which included 15 patients that underwent total hip replacements and 9 patients following hip resurfacings. These patients were followed prospectively with the Harris Hip Score (HHS) and the University of California Los Angeles (UCLA) activity score. Serial radiographs and ion levels were analyzed at regular intervals. Oxidative stress markers (total anti-oxidants, peroxide, and nitro-tyrosine) were also measured from whole blood to determine if these correlated with an increase ion levels in outlier patients.

Results: Post-operative HHS and UCLA activity scores improved significantly compared to pre-operative values. There was no statistical correlation between outlier ion levels, patient demographics, HHS and UCLA activity scores. Radiologic parameters such as cup inclination and femoral component neck-shaft angle could not account for higher ion levels in these outliers. Oxidative stress markers were similar to the levels observed in the control patients with normal ion values following with metal on metal hip implants.

Conclusion: We could not identify any patient or surgical factors that could explain the abnormally high metal ion levels in the outlier patients. This suggests that the cause of ion level increase is multifactorial. The clinical relevance of such high levels of ions remains unknown given that there was no increase in serum oxidative stress markers. Further studies are necessary to better understand the effect of abnormal elevations in metal ions given the recent concerns of pseudotumours following metal on metal hip implants.


Orthopaedic Proceedings
Vol. 93-B, Issue SUPP_II | Pages 130 - 130
1 May 2011
Nikolaou V Khoury V Huk O Petit A Bergeron S Zukor D Antoniou J
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Aim: To determine the MRI findings in patients with persistent painful metal on metal (MOM) hip arthroplasty and compare the results with a control group of patients with MOM or metal on polyethylene THA without symptoms.

Methods: 20 patients with normal inflammatory markers and normal plain radiographic imaging that had undergone primary THA were enrolled to this study. Patients were chosen to be included in 4 groups;

Patients having metal-on-polyethylene THA or resurfacing without pain (Control group),

Patients having MOM THA or resurfacing with high levels of metal ions (cobalt and chromium) and having pain

Patients having MOM THA or resurfacing with high levels of metal ions but having no pain and

Patients having MOM THA or resurfacing with low levels of metal ions and having no pain.

Operated hips were evaluated with MRI by one musculoskeletal radiologist who was blinded to the radiographic findings and clinical symptoms. All images were assessed for the presence of a juxtaarticular or periprosthetic abnormalities, including fluid collections, soft tissue masses, osseous abnormalities, and patterns of contrast enhancement of lesions.

Results: 5 patients were included in each group. All patients had undergone their THA at least 1 year prior to the MRI examination (mean 18 months). MRI findings including muscle atrophies, joint effusions, stress fractures, bone marrow oedema and muscle avulsions were equally distributed in all groups.

Conclusions: MOM THA or high metal ion levels had no specific MRI findings to explain the hip pain in these groups of patients.


Orthopaedic Proceedings
Vol. 92-B, Issue SUPP_IV | Pages 510 - 510
1 Oct 2010
Bergeron S Antoniou J Chakravertty R Ma B Rudan J
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Introduction: Adequate bone in the femoral head and neck is a prerequisite in ensuring the longevity of a surface arthroplasty. The pistol grip deformity is one of the most common bony abnormalities of the femoral head encountered at the time of resurfacing. Severe flattening results in segmental bone loss requiring adjustments in the alignment of the femoral component to achieve optimal orientation. However, very little is known as to how the femoral implant positioning will be affected by increasing deformity. The purpose of this study was to classify the deformity of the femoral head to better understand how it influences the alignment of the femoral component during surface arthroplasty. This classification was then used to determine whether the femoral implant can be safely inserted with optimal alignment despite progressive deformity of the femoral head and neck.

Methods: The classification was developed using plain radiographs and computer tomography scans from 61 patients (66 hips) who presented with primary osteoarthritis prior to hip resurfacing. Surface arthroplasty simulations were generated with three-dimensional computed tomography to quantify the change in femoral component orientation from the neutral position that would allow optimal alignment. The biomechanical parameters were also calculated to determine the influence of the deformity on the final implant position.

Results: There were 47 men and 14 women, with a mean age of 50.3 years (range, 33 to 63 years). Three categories of femoral head deformity were created using a modified femoral head ratio (Normal ≥0.9, Mild = 0.75 – 0.9 and Severe < 0.75). There were a total of 32 normal hips (48%), 23 hips (35%) with mild deformity and 11 hips (17%) with severe deformity of the proximal femur. A severe deformity required significantly more superior translation of the entry point (p=0.027) and greater reaming depth (p=0.012) to allow safe insertion in relative valgus without notching. This could be achieved while preserving length discrepancy (p=0.17) and minimizing the component-head size difference (p=0.16), although femoral offset was significantly reduced (p=0.025).

Conclusion: A classification of femoral head deformity was created to better understand how progressive deformity influences the alignment of the femoral component during surface arthroplasty. This classification is simple and easily measured using standard AP radiographs of the hip. We found that the femoral component can be safely inserted with optimal alignment during surface arthroplasty by modifying the surgical technique in the face of severe deformity.


Orthopaedic Proceedings
Vol. 92-B, Issue SUPP_I | Pages 4 - 4
1 Mar 2010
Debiparshad K Antoniou J Huk OL Mwale F Zukor DJ Petit A
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Purpose: Metal-on-metal articulations in total hip arthroplasty (THA) have been recently re-introduced for the treatment of osteoarthritis. There have been excellent short-term clinical outcomes reported. The long-term clinical results of these implants are still unknown. In this study, we examined the four to ten -year results of metal-on-metal total hip arthroplasty at our institution.

Method: All patients receiving a metal-on-metal prosthesis were prospectively registered in a computerized database. The period July 1997 – May 2003 was selected for analysis. This group contained 251 cases (207 patients) and had an average follow-up of 6.8 years. Of the 251 implants the femoral head components were either Metasul (203) or Ultamet (48) and the acetubular component Fitek (12), Interop (191) or Pinnacle (48). All patients were assessed with the Harris hip and UCLA activity scores at routinely scheduled follow-up visits. The mean age of patients at surgery was 52 years (18 to 70 years old). The average follow-up was of 6.8 years (4.2–10 years). No patients were lost to follow-up.

Results: The mean UCLA and Harris hip score pre-operatively were 3.6 (1–8) and 42 (15–81), respectively. Values of the UCLA and the Harris hip scores on the most recent follow-up visit were 7.0 (60% of patients had a score equal to or greater than 7) and 85 (70% of patients had a score equal to or greater than 85), respectively. Thirteen hips (5.1%) were revised in total: 1 (0.4%) for early deep infection, 1 (0.4%) for revision of undersized stem and 11 (4.3%) for acetabular revisions due to manufacturing defects. Radiological results showed no osteolysis and 9 non-progressive radiolucency.

Conclusion: At four to ten year follow-up, metal-on-metal total hip replacement provides consistently good clinical and radiographic results with a low revision rate. They provide longstanding pain alleviation and improved function and activity levels in patients suffering from osteoarthritis.