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Background aims The discovery of regenerative and immunosuppressive capacities of mesenchymal stromal cells (MSCs) raises hope for patients with tissue-damaging or severe, treatment-refractory autoimmune disorders. We previously presented a method to expand human MSCs in a bioreactor under standardized Good Manufacturing Practice conditions. Now we characterized the impact of critical treatment conditions on MSCs with respect to immunosuppressive capabilities and proliferation. Methods MSC proliferation and survival after γ irradiation were determined by 5-carboxyfluorescein diacetate N-succinimidyl ester and annexinV/4′,6-diamidino-2-phenylindole (DAPI) staining, respectively. T-cell proliferation assays were used to assess the effect of γ irradiation, passaging, cryopreservation, post-thaw equilibration time and hypoxia on T-cell suppressive capacities of MSCs. Quantitative polymerase chain reaction and β-galactosidase staining served as tools to investigate differences between immunosuppressive and non-immunosuppressive MSCs. Results γ irradiation of MSCs abrogated their proliferation while vitality and T-cell inhibitory capacity were preserved. Passaging and long cryopreservation time decreased the T-cell suppressive function of MSCs, and postthaw equilibration time of 5 days restored this capability. Hypoxic culture markedly increased MSC proliferation without affecting their T-cell-suppressive capacity and phenotype. Furthermore, T-cell suppressive MSCs showed higher CXCL12 expression and less β-galactosidase staining than non-suppressive MSCs. Discussion We demonstrate that γ irradiation is an effective strategy to abrogate MSC proliferation without impairing the cells' immunosuppressive function. Hypoxia significantly enhanced MSC expansion, allowing for transplantation of MSCs with low passage number. In summary, our optimized MSC expansion protocol successfully addressed the issues of safety and preservation of immunosuppressive MSC function after ex vivo expansion for therapeutic purposes.

Purpose The aim of this study was to assess the reproducibility of using magnetic resonance imaging (MRI) to analyze the preserved anatomic knee compartments following unicondylar knee arthroplasty (UKA) with zirconium femoral components. It was hypothesized that evaluation of the cartilage, ligaments, meniscus, and tendons would result in a high rate of inter-observer reliability. Scope Ten patients underwent MRI of the knee tailored to reduce metallic artifact following medial UKA with zirconium femoral implants. Cartilage, external meniscus, collateral and cruciate ligaments, the quadriceps and patellar tendons, and the presence of joint effusion were evaluated by two independent investigators. The reviewers provided degrees of confidence with their evaluation of each parameter through the use of a five-point scale. Inter-observer agreement was calculated and inter-observer reliability was determined by use of Cohen's Kappa. Artifacts originating from the implants were rarely observed. There was excellent inter-observer reliability (i.e., high Cohen's Kappa) for all assessed structures, and a high level of observer confidence for the evaluation of the cartilage, meniscus, tendons, ligaments, and joint effusion. Conclusion In this study tailored MRI allows for reproducible analysis of the preserved knee joint compartment after UKA for zirconium implants. This technique might prove helpful in the assessment of painful knee joints after UKA with other metallic materials as new MRI software programs, which suppress metal artifacts, are developed.

Ultrasonography of the hip has gained wide acceptance as reliable method for diagnosing developmental hip dysplasia in infants. This blinded study was conducted to investigate the influence of two linear probes with different frequency on diagnostic accuracy and reproducibility. In 206 consecutive newborns, the bony roof angle (α-angle) and cartilage roof angle (β-angle) were assessed by three investigators according to Graf. The hips were measured twice both with a 7.5 MHz and a 12 MHz linear transducer (Sonoline G60S ® ultrasound system; Siemens, Erlangen, Germany). Each investigator evaluated his sonograms 6 weeks later. Depending on the mean values for α-angle and β-angle, the hips were classified. Reproducibility of the Graf classification was not found to be influenced. In two of three investigators, we observed significant improvement concerning variation of the β-angle with the 12 MHz probe. The use of high-resolution transmitters may improve diagnostic accuracy in ultrasonography of the newborn’s hip.

Grice-Green extra-articular subtalar arthrodesis is a treatment option for advanced valgus rearfoot deformity of neuromuscular origin in young patients. The purpose of this study was to evaluate long-term results of the procedure in ambulatory cerebral palsy patients. From January 1975 to December 1993, 57 operations were performed in 39 patients (22 males, 17 females) with a mean age of 7.8 ± 2.7 (range 3.9–14.4) years for excessive symptomatic hindfoot valgus. Thirty-five patients (51 feet) were followed for a mean 22.6 ± 4.6 (range 16.0–32.3) years. Preoperative GMFCS score, ambulation, hindfoot position, podoscopic view of the weight-bearing feet, and radiological examinations, along with complications and orthotic use, were compared. Based on a clinical rating scale, 39 results were excellent or good, 8 were fair, and 4 were poor. The poor results were attributed to hindfoot valgus recurrence or varus overcorrection. One case needed revisional surgery owing to slippage of the graft. Overall, the GMFCS score, level of ambulatory distance, and use of foot orthoses improved. Grice-Green subtalar arthrodesis did not result in early degenerative changes at the midterm follow-up. The procedure offers safe and long-lasting correction for severe and symptomatic hindfoot disorders in patients with cerebral palsy. Together with physiotherapy and orthotic use, it can improve ambulation. Whereas a slight hindfoot valgus in patients with cerebral palsy is tolerable without lack of function, overcorrection should be avoided.

Evaluation of gait performance and muscle activity patterns as well as clinical efficacy and safety after intraarticular injections with hyaluronan (Ostenil) compared with triamcinolone (Volon A10) injections in patients with knee osteoarthritis. This was a prospective, randomised, double-blind clinical trial evaluating the influence of five injections of hyaluronan or triamcinolone on gait pattern and muscle activity. For the clinical evaluation visual analogue scale, Lequesne index, and Knee Society Score were used. Quality of life was estimated with the SF-36. The definitive analysis was performed on the population who received all five injections and were examined in the two follow-up visits. Fifteen patients were treated with triamcinolone and 20 with hyaluronan. Significant improvement could be demonstrated for pain scale and clinical scores in both groups. Gait patterns showed significant differences only for Knee Abduction Moment (p=0.007) in the hyaluronan group and for Maximum Vertical Force 1 and 2 between the both groups in the follow up visit (p=0.018) (p=0.019). In both groups there was no significant difference regarding to muscle activity. Quality of life showed no changes in all visits between the groups. Hyaluronan was not superior to triamcinolone. The results suggest that treatment with hyaluronan can reduce pain and improve knee function. A significant short term improvement in gait and muscle activity patterns, however, was not observed, in either hyaluronan or triamcinolone.