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Orthopaedic Proceedings
Vol. 106-B, Issue SUPP_19 | Pages 64 - 64
22 Nov 2024
Mbuku RB Poilvache H Van Bambeke F Cornu O
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Aim

The management of PJIs is slowed down by the presence of bacteria forming biofilms where they may withstand antibiotic therapy. The use of adjuvant strategies, such as hydrolytic enzymes cocktail targeting biofilm matrices and facilitating their dispersion, is a promising option to limit impact of biofilms. Our aim was to evaluate the effect of enzymes cocktail combined with antibiotic dual therapy of rifampicin and vancomycin in a relevant in-vitro model.

Method

Mature methicillin-resistant Staphylococcus aureus biofilms were grown on Ti-6Al-4V coupons by adding 1mL of a 8Log10 ATCC 33591 suspension in TGN (TSB + 1% glucose + 2% NaCl) to 24-wells plates containing the coupons and incubating the plates for 24h at 37°C with a continuous 50rpm agitation. The samples were rinsed and placed in 6 wells plates containing 1ml of the enzymatic cocktail (C.D.D.) solution (tris-buffered (pH 7.0) solution of 400 U/ml of aspecific DNA/RNA endonuclease, 50 U/ml of endo-1,4-b-D-glucanase, and 0.06 U/ml of β-N-acetylhexosaminidase). 9ml of TGN or TGN containing antibiotics RIF/VAN (rifampicin 5µg/mL + vancomycin 8µg/mL) at clinically relevant concentrations found locally in bone or joints, was then added and the samples were incubated in identical conditions for 24h. The samples were then recovered and rinsed. CFU counts were obtained by recovering the bacteria with sonication, serial dilutions, and TSA plating. Biomass was determined via crystal violet staining, followed by dye solubilization in acetic acid, and absorbance measurement using a spectrophotometer.


Orthopaedic Proceedings
Vol. 106-B, Issue SUPP_19 | Pages 86 - 86
22 Nov 2024
Lentini A Djoko J Putineanu D Tribak K Coyette M Yombi J Cornu O
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Aim

Bone infections often manifest with soft tissue complications such as severe scarring, fistulas, or ulcerations. Ideally, their management involves thorough debridement of infected bone and associated soft tissues, along with achieving stable bone structure, substantial tissue coverage, and long-term antibiotic therapy. The formation of a multidisciplinary team comprising orthopedic surgeons, plastic surgeons, and infectious disease specialists is essential in addressing the most complex cases.

Method

We conducted a retrospective study during six years (2018-2023) at our university center. Focusing on the most challenging cases, we included patients with bone infections in the leg and/or foot requiring free flap reconstruction. Each patient underwent simultaneous bone debridement and reconstruction by the orthopedic team, alongside soft tissue debridement and free flap reconstruction by the plastic surgery team. Targeted antibiotic therapy for either 6 weeks (acute) or 12 weeks (chronic osteitis) was initiated based on intraoperative cultures. Additional procedures such as allografts, arthrodesis, or autografts were performed if necessary. We analyzed the rates of bone union, infection resolution, and limb preservation.


Orthopaedic Proceedings
Vol. 106-B, Issue SUPP_18 | Pages 110 - 110
14 Nov 2024
Vettese J Julie M Evrard R Fieve L Lengelé B Cornu O
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Introduction

In tissue engineering, the establishment of sufficient vascularization is essential for tissue viability and functionality. Inadequate vascularization disrupts nutrients and oxygen supply. Nonetheless, regenerating intricate vascular networks represents a significant challenge. Consequently, research efforts devoted to preserving and regenerating functional vascular networks in engineered tissues are of paramount importance. The present work aims to validate a decellularisation process with preservation of the vascular network and extracellular matrix (ECM) components in fasciocutaneous flaps.

Method

Five vascularized fasciocutaneous flaps from cadaveric donors were carefully harvested from the anterolateral thigh (ALT), preserving the main perforator of the fascia lata. The entire ALT flap underwent decellularization by perfusion using a clinically validated chemical protocol. Fluoroscopy and computed tomography (CT) were used to analyze the persistence of the vascular network within the flap, pre- and post-decellularization. Histological analysis, including hematoxylin and eosin staining, and quantitative DNA assessment evaluated decellularization efficacy. Further qualitative (immunohistochemistry, IHC) and quantitative analyses were conducted to assess the preservation of ECM components, such as collagen, glycosaminoglycans, and elastin.


Orthopaedic Proceedings
Vol. 106-B, Issue SUPP_2 | Pages 125 - 125
2 Jan 2024
Mbuku R Sanchez C Evrard R Englebert A Manon J Henriet V Nolens G Duy K Schubert T Henrotin Y Cornu O
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To design slow resorption patient-specific bone graft whose properties of bone regeneration are increased by its geometry and composition and to assess it in in-vitro and in-vivo models.

A graft composed by hydroxyapatite (HA) and β-TCP was designed as a cylinder with 3D gyroid porosities and 7 mm medullary space based on swine's anatomy. It was produced using a stereolithography 3D-printing machine (V6000, Prodways).

Sterile bone grafts impregnated with or without a 10µg/mL porcine BMP-2 (pBMP-2) solution were implanted into porcine femurs in a bone loss model. Bone defect was bi-weekly evaluated by X-ray during 3 months. After sacrifice, microscanner and non-decalcified histology analysis were conducted on biopsies.

Finally, osteoblasts were cultured inside the bone graft or in monolayer underneath the bone graft. Cell viability, proliferation, and gene expression were assessed after 7 and 14 days of cell culture (n=3 patients).

3D scaffolds were successfully manufactured with a composition of 80% HA and 20% β-TCP ±5% with indentation compressive strength of 4.14 MPa and bending strength of 11.8MPa.

In vivo study showed that bone regeneration was highly improved in presence of pBMP-2. Micro-CT shows a filling of the gyroid sinuses of the implant (Figure 1).

In vitro, the presence of BMP2 did not influence the viability of the osteoblasts and the mortality remained below 3%. After 7 days, the presence of BMP2 in the scaffold significantly increased by 85 and 65% the COL1A1 expression and by 8 and 33-fold the TNAP expression by osteoblasts in the monolayer or in the scaffold, respectively. This BMP2 effect was transient in monolayer and did not modify gene expression at day 14.

BMP2-impregnated bone graft is a promising patient-personalized 3D-printed solution for bone defect regeneration, by promoting neighboring host cells recruitment and solid new bone formation.

For any figures and tables, please contact the authors directly.


Orthopaedic Proceedings
Vol. 106-B, Issue SUPP_2 | Pages 124 - 124
2 Jan 2024
Manon J Evrard R Fievé L Xhema D Maistriaux L Schubert T Lengelé B Behets C Cornu O
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Decellularization techniques have advanced to reduce the risk of immune rejection in transplantation. Validation of these protocols typically relies on Crapo's criteria1, which include the absence of visible nuclei and low DNA content. In our study, five decellularization protocols were compared to determine the optimal approach for human fascia lata (HFL) samples. However, our findings raised questions as to why recipients can still develop immunity despite meeting validation criteria.

HFL samples were decellularized using four protocols with SDS-Triton X100-DNase (D1 to D4-HFL) and one protocol using solvent-detergent-based baths (D5-HFL). The decellularized samples (D-HFL) were compared to native samples (N-HFL) using histology, and DNA content was measured. The human leukocyte antigen (HLA) content within the matrix was assessed using western blot analysis. Both D-HFL and N-HFL samples, along with negative control patches, were implanted in the backs of 28 Wistar rats. Anti-human IgG serum levels were evaluated after one month.

H&E and Hoechst staining revealed the absence of residual cells in all decellularization protocols. DNA content was consistently below the critical threshold (p<0.05). All implanted D-HFL samples resulted in significantly lower anti-human IgG levels compared to N-HFL (p<0.01). However, 2.5 out of 4 rats developed immunity after being implanted with D1 to D4-HFL, with varying levels of anti-human IgG. Only rats implanted with D5-HFL showed undetectable levels of IgG and were considered non-immunized. Western blot analysis indicated that only D5-HFL had a residual HLA content below 1%.

The literature on decellularization has primarily relied on Crapo's criteria, which do not consider the role of HLA mismatch in acute immune rejection. Our results suggest that a residual HLA content below 1% should also be considered to prevent immunization, even if other validation criteria are met. Further research is needed to evaluate the impact of residual HLA levels on human allotransplantation outcomes.


Orthopaedic Proceedings
Vol. 105-B, Issue SUPP_17 | Pages 45 - 45
24 Nov 2023
Dendoncker K Putzeys G Cornu O Nieuwenhuizen T Bertrand M Valster H Croes K
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Aim

Local antibiotics released through a carrier is a commonly used technique to prevent infection in orthopaedic procedures. An interesting carrier in aseptic bone reconstructive surgery are bone chips impregnated with AB solution. Systemically administered Cefazolin (CFZ) is used for surgical site infection prophylaxis however in vitro study showed that fresh frozen and processed bone chips impregnated with CFZ solution completely release the CFZ within a few hours. On the other hand irradiated freeze-dried bone chips, treated with supercritical CO2 (scCO2) have been shown to be an efficient carrier for the antibiotics vancomycine or tobramycine.

With this pilot study we wanted to investigate if CFZ solution impregnation of bone chips treated with scCO2 shows a more favorable release pattern of CFZ.

Method

The bone chips were prepared using the standard scCO2 protocol and were impregnated with 100 mg/ml cefazolin at different timepoints during the process: before freeze drying (BC type A), after freeze drying (BC type B) and after gamma-irradiation. 0.5g of the impregnated bone grafts were incubated with 5ml of fetal calf serum (FCS) at 37°C. At 2, 4, 6, 8 and 24h of incubation 200µl of eluate was taken for analysis. After 24h the remaining FCS was removed, bone grafts were washed and new FCS (5ml) was added. Consecutive eluate samples were taken at 48, 72 and 96h of incubation.

The concentration of CFZ in the eluates was measured with the validated UPLC-DAD method. Analysis was performed in triplicate.


Orthopaedic Proceedings
Vol. 105-B, Issue SUPP_7 | Pages 40 - 40
4 Apr 2023
Evrard R Maistriaux L Manon J Rafferty C Cornu O Gianello P Lengelé B Schubert T
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The purpose of this study is to enhance massive bone allografts osseointegration used to reconstruct large bone defects. These allografts show >50% complication rate requiring surgical revision in 20% cases. A new protocol for total bone decellularisation exploiting the vasculature can offer a reduction of postoperative complication by annihilating immune response and improving cellular colonization/ osseointegration.

The nutrient artery of 18 porcine bones - humerus/femur/radius/ulna - was cannulated. The decellularization process involved immersion and sequential perfusion with specific solvents over a course of one week. Perfusion was realized by a peristaltic pump (mean flow rate: 6ml/min). The benefit of arterial perfusion was compared to a control group kept in immersion baths without perfusion.

Bone samples were processed for histology (HE, Masson's trichrome and DAPI for cell detection), immunohistochemistry (IHC : Collagen IV/elastin for intraosseous vascular system evaluation, Swine Leukocyte Antigen – SLA for immunogenicity in addition to cellular clearance) and DNA quantification. Sterility and solvent residues in the graft were also evaluated with thioglycolate test and pH test respectively.

Compared to native bones, no cells could be detected and residual DNA was <50ng/mg dry weight. Intramedullary spaces were completely cleaned. IHC showed the preservation of intracortical vasculature with channels bounded by Collagen IV and elastin within Haversian systems. IHC also showed a significant decrease in SLA signaling. All grafts were sterile at the last decellularization step and showed no solvent residue. The control group kept in immersion baths, paired with 6 perfused radii/ulnae, showed that the perfusion is mandatory to ensure complete decellularisation.

Our results prove the effectiveness of a new concept of total bone decellularisation by perfusion. These promising results could lead to a new technique of Vascularized Composite Allograft transposable to pre-clinical and clinical models.


Orthopaedic Proceedings
Vol. 104-B, Issue SUPP_10 | Pages 38 - 38
1 Oct 2022
Coppeaux M Popescu D Julie M Poilvache H Mbuku RB Maïte VC Yombi J Cornu O
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Introduction

The surgical management of late PJI was usually done in two stages with the placement of a temporary cement spacer. The development of one-stage surgical care raises questions about the two-stage strategy. The objectives of this study are to identify the complications related to the presence of the cement spacer within a two-stage strategy. The septic recurrence rate is also evaluated after a minimum follow-up of two years.

Material and methods

Medical files of 208 patients (101 knees and 107 hips) who underwent a two-stage revision for late PJI prosthesis infection were retrospectively reviewed. Antibiotic loaded articulated homemade cement spacers were used. Second stage was usually planned on average 4 to 6 weeks after the first stage. Patients were allowed to walk without loading. The success rate was defined as the absence of septic recurrence after a minimum follow-up of two years. Descriptive statistics and uni- and multivariate analysis were conducted.


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 74 - 74
1 Dec 2019
Pastor I Poilvache H Morcillo D van Cauter M Rodriguez-Villalobos H Yombi J Cornu O
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Aim

We wonder what the results of two stage procedures were in terms of morbidity (amputation, dead) and infection recurrence. We also seek to identify risk factors for failure and see if the results of a second two stage surgery were not even worse.

Material and Methods

We retrospectively reviewed 140 prosthetic joint infection (PJI) treated with a two stage procedure. Patient data has been reviewed to determine which factors would be predictive for failure.


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 38 - 38
1 Dec 2019
Kouamé JK Manon J Fonkoue L Kodo M Detrembleur C Cornu O
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Introduction

The management of patients with open leg fracture in Ivory Coast does not meet the standards of developed countries due to socio-economic conditions, accessibility and organization of care. However, is this care problematic? Is it associated with more post-traumatic infection or mechanical complications and are these correlated with the delay for treatment and the method of treatment?

Material and Methods

This is a single-center prospective study conducted on between January 2018 and May 2018 at Bouaké University Hospital. The observed parameters included factors related to patient, fracture and treatment conditions and were correlated with the rates of complications by multivariate analysis.


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 52 - 52
1 Dec 2019
Yildiz H Cornu O d'Abadie P Yombi J
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Aim

Staphylococcus aureus bacteremia (SAB) is associated with significant morbidity and mortality, 20–30 % risk of infection in patient with implant related infection (IRI) .18F-FDG PET/CT is helpful in the management of SAB, leading to detection of more metastatic foci and treatment modification and finally decrease relapses and mortality rate. Our objective was to analyze mortality in high risk SAB patients undergoing 18F-FDG PET/CT and to see whether it's use in patients with IRI reduced their mortality.

Method

We performed a retrospective study at a university hospital in Belgium. All cases of high risk adult SAB between January 2014 and June 2017 were reviewed. We collected the clinical characteristics including presence of metastatic foci on 18F-FDG PET/ CT, mortality at 1 year.


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 45 - 45
1 Dec 2019
Huard M Detrembleur C Poilvache H van Cauter M Driesen R Yombi J Neyt J Cornu O
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Aim

Apart from other biomarkers isolated in the synovial fluid, alpha-defensin appears to be a promising diagnostic tool to confirm a periprosthetic joint infection (PJI) in the hip or knee. The purpose of this study was to evaluate the sensitivity and specificity of an alpha defensin lateral flow (ADLF) test compared to usual standard classifications in the diagnostic management of PJI.

Method

This investigation was set up as a multicenter prospective cohort study. Synovial fluid was obtained by means of joint aspiration or intra-operative tissue biopsies. A presumptive PJI diagnosis was made according to criteria outlined by the Musculoskeletal Infection Society (MSIS), the Infectious Diseases Society of America (IDSA) and the European Bone and Joint Infection Society (EBJIS). The intention to treat by the surgeon was logged. Sensibility and specificity for the ADLF test was plotted for each aforementioned diagnostic algorithm. Spearman correlations between all scores were analyzed. Multiple logistic regression was used to determine the contribution of independent variables to the probability of PJI.


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 35 - 35
1 Dec 2019
Orioli L Hammer F Berg BV Putineanu DC Maiter D Yombi J Cornu O Bernard V
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Aim

Our study aimed to analyze 1) the prevalence of peripheral arterial disease (PAD) and infection in diabetic patients with and without Charcot foot (CF), 2) the characteristics of PAD in these 2 groups, 3) the prognosis of patients with CF and PAD and/or infection.

Method

We retrospectively reviewed the medical and radiological records of 172 hospitalized patients in our diabetic foot unit between 2010 and 2014. These patients were identified using the ICD-9-CM. The CF group and the diabetic foot (DF) group included 56 and 116 patients, respectively. All statistical analyses were performed using SPSS 25.0.01. A p <0.05 was considered as statistically significant.


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 23 - 23
1 Dec 2019
Poilvache H Ruiz-Sorribas A Rodriguez-Villalobos H Sakoulas G Cornu O van Bambeke F
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Aim

Irrigation is a major step during debridement surgery in the context of Prosthetic Joint Infections (PJI), but its effects on biofilms are poorly described.

The present study aims at evaluating the effect of PW alone or followed by antibiotics on MSSA and MRSA biofilms grown on Ti6Al4V coupons in-vitro.

Method

Strains: 1 reference (MSSA: ATCC25923; MRSA: ATCC33591) and 2 clinical MSSA and MRSA isolated from PJI.

Biofilm culture: Coupons were incubated for 24h at 37°C with bacteria (starting inoculum ∼6.6Log10CFU/mL in TGN [TSB + 1% glucose + 2% NaCl]), under shaking at 50rpm.

Treatment: Half of the coupons were irrigated with 50mL physiological serum from 5cm using a Stryker Interpulse; the coupons were then either analysed (ControlT0 and PWT0) or reincubated for 24h in TGN or TGN containing flucloxacillin (MSSA) or vancomycin (MRSA) at MIC or 20mg/L.

Analysis: Coupons were rinsed twice with PBS. Biomass was measured by crystal violet (CV) assay. CFUs were counted after recovering bacteria from coupons using sonication and TSA plating.


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 8 - 8
1 Dec 2019
Morcillo D Pastor I Detrembleur C Poilvache H van Cauter M Yombi J Cornu O
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Background

DAIR is an attractive treatment for PJI. The purpose of this study is to determine predictive factors of failure.

Materials and Methods

We reviewed all DAIR procedures for hip PJI performed between 2002–2017 (n=69). Data recorded included all factors correlated with treatment failure. KLIC score and an adapted McPherson score (sum of three criteria where the lower score is three for 3 A and the higher is 9 for 3 C) were analyzed.


The Bone & Joint Journal
Vol. 101-B, Issue 9 | Pages 1122 - 1128
1 Sep 2019
Yombi JC Putineanu DC Cornu O Lavand’homme P Cornette P Castanares-Zapatero D

Aims

Low haemoglobin (Hb) at admission has been identified as a risk factor for mortality for elderly patients with hip fractures in some studies. However, this remains controversial. This study aims to analyze the association between Hb level at admission and mortality in elderly patients with hip fracture undergoing surgery.

Patients and Methods

All consecutive patients (prospective database) admitted with hip fracture operated in a tertiary hospital between 2012 and 2016 were analyzed. We collected patient characteristics, time to surgery, duration and type of surgery, comorbidities, Hb at admission, nadir of Hb after surgery, the use and amount of red blood cells (RBCs) transfusion products, postoperative complications, and death. The main outcome measures were mortality at 30 days, 90 days, 180 days, and one year after surgery.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 73 - 73
1 Dec 2017
Pierret F Migaud H Loiez C Valette M Beltrand E Yombi J Cornu O Senneville E Cauter MV
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Aim

The treatment of a chronic prosthetic joint infection (PJI) is a combination of the bacteria's identification, a «carcinological» surgery and an appropriate antibiotherapy. In case of gram positive cocci infection, rifampicin is often used.

The aim of this study is to determine which factors are responsible for the development of resistance to rifampicine.

Method

All patients had a total hip (THA) or knee (TKA) arthroplasty with a chronic infection. They were treated with a two-time surgery. All of them received a bi-antibiotic treatment. In case of gram positive cocci infection, and according to the susceptibility test, they received rifampicin. The 221 patients were operated from July 1997 to November 2013 in 3 university centers (one Belgian and two French) and were retrospectively analysed. The demographical, clinical and bacterial data as well as the antibiotic treatment were collected. The healing was defined as the absence of recurrence during the 2 years following surgery.


The Bone & Joint Journal
Vol. 99-B, Issue 9 | Pages 1176 - 1182
1 Sep 2017
Berger P Van Cauter M Driesen R Neyt J Cornu O Bellemans J

Aims

The purpose of this current multicentre study is to analyse the presence of alpha-defensin proteins in synovial fluid using the Synovasure lateral flow device and to determine its diagnostic reliability and accuracy compared with the prosthetic joint infection (PJI) criteria produced by the Musculoskeletal Infection Society (MSIS).

Patients and Methods

A cohort of 121 patients comprising 85 total knee arthroplasties and 36 total hip arthroplasties was prospectively evaluated between May 2015 and June 2016 in three different orthopaedic centres. The tests were performed on patients with a chronically painful prosthesis undergoing a joint aspiration in a diagnostic pathway or during revision surgery.


Orthopaedic Proceedings
Vol. 98-B, Issue SUPP_23 | Pages 30 - 30
1 Dec 2016
Cornu O Van Cauter M Kaminski L Jean-Cyr Y Rodriguez-Villalobos H
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Aim

Bacterial identification in musculoskeletal infection is sometimes difficult and treatment strategy difficult facing unknown pathogen agent. We wonder if the delay of incubation and the preservation conditions of the samples between surgical procurement and subculture on plates have an influence.

Method

25 cm³ bone fragments were obtained from femoral heads retrieved during hip arthroplasty and excluded for bone transplant donation. Informed consent was obtained from the donor for research purpose. The study was approved by the Ethic Committee (N°B403201317725). Bone fragments were immersed for 30 minutes under gently agitation (140 RPM) at 35°C in a physiologic solution (negative control) or two solutions with two concentrations of staphylococcus epidermidis (0.5 Mc Farland or 1.5× 108 bacteria and 7.5×102 bacteria). Bone samples were separated and preserved at room temperature or at 4°C until seeded on Petri Plates to observe the influence of preservation conditions. Samples were plated after different delays (T0, T30min, T1H, T2H, T4H, T6H, T8H, T12H, T16H, T24H et T48H) to observe the influence of delay of culture. Experiments were repeated 5 times. When culture was positive, results were expressed with the number of colony.


Orthopaedic Proceedings
Vol. 98-B, Issue SUPP_4 | Pages 66 - 66
1 Jan 2016
Thienpont E Schwab P Forthomme JP Cornu O
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Introduction

Patient satisfaction after total hip arthroplasty (THA) has been reported to be significantly better than after total knee arthroplasty (TKA). The same has been observed for the capacity to forget during daily life activities about the operated joint. Recently a new patient reported outcome score, the Forgotten Joint Score (FJS-12) a twelve item questionaire, has been used to evaluate postoperative outcome in joint arthroplasty. A better FJS-12 score was measured in THA than in TKA objectivating the intuitive feeling that the joint was more forgotten in THA than in TKA.

Hypothesis

A higher preoperative FJS-12 score is the reason for a higher postoperative FJS-12 score in THA compared to TKA