Stéphane Boisgard

Mesenchymal content of fresh bone marrow: a proposed quality control method for cell therapy.

The scarcity of mesenchymal stem cells (MSC) in bone marrow (BM) has justified their ex vivo expansion before therapeutic use, but a method to evaluate the quality of initial mesenchymal content and track the modifications induced by graft processing has not yet been proposed. The aim of this study was to establish such a procedure. Flow cytometric and functional assay methods were modified to count CD45− CD14−/CD73+ subsets containing all MSC and used them to study BM from spongy bone (SB) and iliac crest aspirate (ICA). These methods detected the target subsets in all BM suspensions derived from SB (n = 154) and ICA, (n = 44) with a satisfactory correlation between immuno‐phenotyping and functional tests by low‐density plating. We noted a higher overall MSC frequency in SB cell suspensions but a lower plating efficiency of CD45− CD14−/CD73+ SB cells under standard culture conditions.

Determinants of satisfaction 1 year after total hip arthroplasty: the role of expectations fulfilment

BackgroundBetween 7% and 15% of patients are dissatisfied after total hip arthroplasty (THA). To assess predictors and postoperative determinants of satisfaction and expectation fulfilment one year after (THA).MethodsBefore THA surgery, 132 patients from three tertiary care centres and their surgeons were interviewed to assess their expectations using the Hospital for Special Surgery Total Hip Replacement Expectations Survey (THR survey). One year after surgery, patients (n = 123) were contacted by phone to complete a questionnaire on expectation fulfilment (THR survey), satisfaction, functional outcome (Womac), and health-related quality of life (SF 12). Univariate and multivariate analyses were performed.ResultsPreoperative predictors of satisfaction were a good mental wellbeing (adjusted OR 1.09 [1.02; 1.16], p = 0.01) and optimistic surgeons expectations (1.07 [1.01; 1.14], p = 0.02). The main postoperative determinant of satisfaction was the fulfilment of patient’s expectations (1.08 [1.04; 1.12], p < 0.001). Expectation fulfilment could be predicted before surgery by young age (regression coefficient −0.55 [−0.88; -0.21], p = 0.002), good physical function (−0.96 [−1.82; -0.10], p = 0.03) and good mental wellbeing (0.56 [0.14; 0.99], p = 0.01). Postoperative determinants of expectation fulfilment were functional outcome (−2.10 [−2.79; -1.42], p <0.001) and pain relief (−14.83 [−22.38; -7.29], p < 0.001).ConclusionTo improve patient satisfaction after THA, patients’ expectations and their fulfilment need to be carefully addressed. Patients with low mental wellbeing or physical function should be identified and specifically informed on expected surgical outcome. Surgeons’ expectations are predictive of satisfaction and information should aim to lower discrepancy between surgeons’ and patients’ expectations.

Complex primary total hip arthroplasty.

Although total hip arthroplasty is now a classic procedure that is well controlled by orthopedic surgeons, some cases remain complex. Difficulties may be due to co-morbidities: obesity, skin problems, muscular problems, a history of neurological disease or associated morphological bone deformities. Obese patients must be informed of their specific risks and a surgical approach must be used that obtains maximum exposure. Healing of incisions is not a particular problem, but adhesions must be assessed. Neurological diseases may require tenotomy and the use of implants that limit instability. Specific techniques or implants are necessary to respect hip biomechanics (offset, neck-shaft angle) in case of a large lever arm or coxa vara. In case of arthrodesis, before THA can be performed, the risk of infection must be specifically evaluated if the etiology is infection, and the strength of the gluteal muscles must be determined. Congenital hip dysplasia presents three problems: the position and coverage of the cup, placement of a specific or custom made femoral stem, with an osteotomy if necessary, and finally lowering the femoral head into the cup by freeing the soft tissues or a shortening osteotomy. Acetabular dysplasia should not be underestimated in the presence of significant bone defect (BD), and reconstruction with a bone graft can be proposed. Sequelae from acetabular fractures presents a problem of associated BD. Internal fixation hardware is rarely an obstacle but the surgical approach should take this into account. Treatment of acetabular protrusio should restore a normal center of rotation, and prevent recurrent progressive protrusion. The use of bone grafts and reinforcement rings are indispensible. Femoral deformities may be congenital or secondary to trauma or osteotomy. They must be evaluated to restore hip biomechanics that are as close to normal as possible. Fixation of implants should restore anteversion, length and the lever arm. Most problems that can make THA a difficult procedure may be anticipated with proper understanding of the case and thorough preoperative planning.

Glucocerebrosidase deficiency dramatically impairs human bone marrow haematopoiesis in an in vitro model of Gaucher disease

One of the cardinal symptoms of type 1 Gaucher Disease (GD) is cytopenia, usually explained by bone marrow (BM) infiltration by Gaucher cells and hypersplenism. However, some cases of cytopenia in splenectomized or treated patients suggest possible other mechanisms. To evaluate intra‐cellular glucocerebrosidase (GlcC) activity in immature progenitors and to prove the conduritol B epoxide (CBE)‐induced inhibition of the enzyme, we used an adapted flow cytometric technique before assessing the direct effect of GlcC deficiency in functional assays. Among haematopoietic cells from healthy donors, monocytes showed the highest GlcC activity but immature CD34+ and mesenchymal cells also had significant GlcC activity. CBE greatly inhibited the enzyme activity of all cell categories. GlcC‐deficient CD34+ cells showed impaired ability to proliferate and differentiate in the expansion assay and had lower frequency of erythroid burst‐forming units, granulocyte colony‐forming units (CFU) and macrophage CFU progenitors, but the effect of GlcC deficiency on megakaryocyte CFU lineage was not significant. GlcC deficiency strongly impaired primitive haematopoiesis in long‐term culture. Furthermore, GlcC deficiency progressively impaired proliferation of mesenchymal progenitors. These data suggest an intrinsic effect of GlcC deficiency on BM immature cells that supplements the pathophysiology of GD and opens new perspectives of therapeutic approach.

The chronic lymphocytic leukemia clone disrupts the bone marrow microenvironment.

The systematic localization of chronic lymphocytic leukemia (CLL) B-cells in the bone marrow (BM), together with the ex vivo protective effect of stromal cells on their spontaneous apoptosis, both indicate a specific role of the BM microenvironment. In vivo, the impact of CLL cells on mesenchymal stromal cells (MSCs) remains a source of debate. Here, we quantified and expanded colony forming unit-fibroblasts (CFU-Fs) from CLL-BM under standard conditions, analyzed the expression of selected genes, and studied secretion profiles. We observed failing of CLL-BM cultures in standard conditions (45.5% vs. <0.1%), and even after adding basic fibroblast growth factor (bFGF), there were fewer CFU-F than from normal BM (1.3 vs. 40/10(6) cells respectively; P<0.01). Furthermore, their polygonal aspect and low proliferative capacity, together with the expression of 384 selected genes and a secreted set of molecules related to senescence-associated secretory phenotype indicated a state of senescence, further confirmed by the higher proportion of senescence-associated β-galactosidase (SA-βGAL)-positive cells and p16INK4a overexpression. In our hands, hypoxic conditions (5% O2) did not rescue CFU-Fs. Given the role of MSC in BM tissue organization, we studied hematons that are generally considered to be elementary BM units. These structures were rare or had even disappeared completely. When hematons were present, we systematically observed nodular B-CLL cell invasion only. These data confirm that the B-CLL clone has a marked impact on MSC and disrupts BM organization in vivo, raising new questions about in vivo pathophysiology.

Assessment of the risk factors for impending fractures following radiotherapy for long bone metastases using CT scan-based virtual simulation: a retrospective study.

BackgroundRadiotherapy for long bone metastases (RTLB) can be complicated by fractures, which considerably increase morbidity and mortality. The aim of this study was to analyze the risk factors for impending fractures following radiotherapy for long bone metastases (RTLB) using CT scan-based virtual simulation.MethodsForty-seven (47) patients were treated with RTLB (18 lung, 11 breast, 10 prostate and 8 other cancers) for a period of 18 months. Two doctors analyzed the CT images prior to radiation therapy. The impending fractures were then monitored and the correlation between bone scan parameters and fracture occurrence was analyzed.ResultsThe male gender ratio was 0.57 and the mean age 62.8 (33–93) years. The average size of the metastatic lesions was 32 (8–87) x 2 (6–81) x 52 (7–408) mm with cortical involvement (CI) in 66% of cases. The site was in the upper third of the bone in 92% of cases (28 femoral, 17 humeral and two tibial).Ten fractures occurred: two during RTLB, seven after one month and one after 6.6 months. The fractured lesions measured 48 (17–87) x 34 (12–66) x 76 (38–408) mm. The predictive parameters for fracture were osteolytic (39% vs. 10%; p = 0.02) and permeative lesions (42% vs. 0%; p < 0.0005), a Mirels score ≥9 (42% vs. 0%; p < 0.0005), circumferential CI ≥30% (71% vs. 0%, p < 0.00001), CI ≥45 mm in height (67% vs. 0%, p < 0.00001) and CI in thickness =100% (40% vs. 0%; p = 0.0008). In the multivariate analysis, circumferential CI ≥30% was the only predictive parameter for fracture (p = 0.00035; OR = 62; CI 95%: 6.5-595). Overall survival was 91% and 40% at one month and twelve months respectively.ConclusionsProphylactic primary fixation surgery should always be considered when the circumferential CI ≥30%.

Randomized blinded trial of standardized written patient information before total knee arthroplasty

Background The effect of patient education before total knee arthroplasty (TKA) is controversial. No consensus exists about the optimal content of educational interventions. In a previous study, we developed and validated an educational booklet on the peri-TKA management of knee osteoarthritis. Purposes Our primary purpose was to evaluate the impact of the educational booklet on knowledge among patients awaiting TKA. Patients and methods This randomized controlled single-blind trial evaluated standard information by the surgeon with or without delivery of the educational booklet 4–6 weeks before primary noncomplex TKA in patients aged 55–75 years with incapacitating knee osteoarthritis. Patients were enrolled at a French surgical center between June 2011 and January 2012. A patient knowledge score was determined at baseline, on the day before TKA, and 3–6 weeks after TKA, using a self-administered questionnaire developed for our previous study. The assessor was blinded to group assignment. Results Of 44 eligible patients, 42 were randomized, 22 to the intervention and 20 to the control group, all of whom were included in the analysis. The groups were comparable at baseline. The intervention was associated with significantly better patient knowledge scores. Conclusions An educational booklet improves knowledge among patients awaiting TKA. A study assessing the impact of the booklet combined with a exercise program would be helpful. Level of evidence Level I, randomized controlled double-blind trial; see S1 CONSORT Checklist. Trial registration clinicaltrials.gov #NCT01747759

Opening-wedge high tibial osteotomy with a secure bone allograft (Osteopure™) and locked plate fixation: Retrospective clinical and radiological evaluation of 69 knees after 7.5 years follow-up

BACKGROUND Medial opening-wedge high tibial osteotomy (OWHTO) provides reliable and long-lasting benefits, despite the wide range of wedge-filling and internal-fixation techniques used. The purpose of this work was to assess the clinical and radiological outcomes in a case-series of OWHTO performed using a secure bone allograft and locked plate fixation. HYPOTHESIS The clinical and radiological outcomes of OWHTO with a high-safety bone allograft and locked plate fixation are similar to those reported in previous case-series studies. MATERIALS AND METHODS A single-centre retrospective design was used to study 69 knees in 64 patients with a mean age of 51.8years (31-53years) and a preoperative hip-knee-ankle (HKA) angle of 173° (165°-180°). The wedge was filled with secure Osteopure™ bone allograft and fixation was achieved using an Integra Surfix(®) locked plate. Mean follow-up was 7.5years (5-9.3years). Clinical and functional outcomes were assessed by determining the IKS and KOOS-PS scores and recording complications related or unrelated to the allograft. The main criterion for assessing OWHTO survival was the time to revision surgery for symptom recurrence. Radiological assessment criteria were the HKA angle, tibial slope, patellar height, and osteoarthritis grade. GESTO criteria were used to evaluate the behaviour of the allograft. RESULTS Of the 69 knees, 64 (92.8%) were re-evaluated. The survival rate after 9.3years was 95%±2.7% (3 failures managed with arthroplasty). The functional IKS score improved significantly, by 20 points (P<0.001). Mean increases of 7.8° for the HKA angle and 3.5° for tibial slope were recorded. Bone healing without radiological abnormalities was consistently achieved within 6months. There were no complications related to the allograft (infections, allergies; local or systemic toxicity). DISCUSSION The clinical, radiological, and safety outcomes documented in our study were similar to those reported in earlier work. LEVEL OF EVIDENCE IV, retrospective case-series study.

Radiographic stereometry for non-metal-backed acetubular cups: 3D wear estimation and related uncertainty

The aim of this work was to assess the 3D wear of non-metal-backed acetubular cups from two conventional digitized radiographs. The centers of the femoral head and the metal ring in the equatorial plane of the cup are located using 3D pose techniques for spheres and circles from sampled points in the images. The method used to locate these points of interest and also estimate their covariance is fully described in previous work. The covariance is used to decrease the bias of the pose estimation, while bootstrapping decreases its variance and gives access to the directions of minimal variability between the two centers for each image. These directions are used to produce the final distance after reconstruction. Results are compared with the 2D technique working on a single anteroposterior (AP) radiograph and assuming null lateral wear. Validation is performed on acetubular cups: (a) simulated by Monte-Carlo, (b) implanted on a pelvic model, and (c) acquired on patients just after arthroplasty. The accuracy in wear for simulation increases from about one to four hundredths of a millimeter as caudal and cranial absolute angulations decreases from 45 degrees to 30 degrees . It is more difficult to assess for real prostheses, but results are shown to lie within the manufacturers dimensional tolerances. Globally, the access to the lateral wear is obtained at the expense of confidence in the global linear wear (0.06 mm), which is doubled with regard to 2D technique but still satisfying for clinical use, whereas the bias of the measurement is decreased.

Collection and reconstruction after harvesting donor tissues from the musculoskeletal system: Technique specific to the lower limbs

The lack of available musculoskeletal grafts in France forces us to import a very large quantity of these tissues to use in complex reconstruction procedures. The goal of this article is to describe methods for collecting donor tissues from the musculoskeletal system and for reconstructing the harvested areas. We also provide a summary of the collection procedures performed, harvested grafts and available tissues. While tissue collection requires a significant time investment, the emergence of dedicated teams may be a solution for increasing the number and quality of human musculoskeletal allograft tissues.