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Avascular necrosis (AVN) is a severe complication of immunosuppressant therapy or chemotherapy. A beneficial AVN therapy with core decompression (CD) and intraosseous infusion of mesenchymal stromal cells (MSCs) has been described in adult patients, but there are only few data on MSC applications in pediatric and young adult patients (PYAP). Between 2006 and 2015, 14 AVN lesions of 10 PYAP (6 females) with a median age of 16.9 years (range 8.5-25.8 years) received CD and intraosseous application of autologous MSCs. Data of these patients were analyzed regarding efficacy, safety, and feasibility of this procedure as AVN therapy and compared with a control group of 13 AVN lesions of 11 PYAP (5 females) with a median age of 17.9 years (range 13.5-27.5 years) who received CD only. During the follow-up analysis [MSC group: median 3.1 (1.6-5.8) years after CD; CD group: median 2.0 (1.5-8.5) years after CD], relative lesion sizes (as assessed by magnetic resonance imaging) compared with the initial lesion volume, were significantly lower (

Background Limited knowledge of stem cell therapies` mechanisms of action hampers their sustainable implementation into the clinic. Specifically, the interactions of transplanted stem cells with the host vasculature and its implications for their therapeutic efficacy are not elucidated. We tested whether adhesion receptors and chemokine receptors on stem cells can be functionally modulated, and consequently if such modulation may substantially affect therapeutically relevant stem cell interactions with the host endothelium. Methods We investigated the effects of cationic molecule polyethylenimine (PEI) treatment with or without nanoparticles on the functions of adhesion receptors and chemokine receptors of human bone marrow-derived Mesenchymal Stem Cells (MSC). Analyses included MSC functions in vitro, as well as homing and therapeutic efficacy in rodent models of central nervous system´s pathologies in vivo. Findings PEI treatment did not affect viability, immunomodulation or differentiation potential of MSC, but increased the CCR4 expression and functionally blocked their adhesion receptors, thus decreasing their adhesion capacity in vitro. Intravenously applied in a rat model of brain injury, the homing rate of PEI-MSC in the brain was highly increased with decreased numbers of adherent PEI-MSC in the lung vasculature. Moreover, in comparison to untreated MSC, PEI-MSC featured increased tumour directed migration in a mouse glioblastoma model, and superior therapeutic efficacy in a murine model of stroke. Interpretation Balanced stem cell adhesion and migration in different parts of the vasculature and tissues together with the local microenvironment impacts their therapeutic efficacy. Funding Robert Bosch Stiftung, IZEPHA grant, EU grant 7 FP Health

Zusammenfassung Hintergrund Die Indikationsstellung zur Operation bezieht ihre Legitimation zum Eingriff in die körperliche Unversehrtheit eines Patienten im Wesentlichen aus dem zu erwartenden Behandlungserfolg. Um die Wahrscheinlichkeit und das Ausmaß eines Operationserfolgs bereits präoperativ abzuschätzen, muss der Chirurg in kurzer Zeit eine Fülle an Parametern evaluieren, die über die rein technische Dimension des Eingriffs hinausgehen. Trotz höchster Versorgungsstandards zeigt die postoperative patientenseitige Zufriedenheit eine immense Streubreite bei für den Arzt technisch zufriedenstellendem Ergebnis. Ziel der folgenden Studie war es, zu untersuchen, mit welcher Präzision individuelle postoperative Zufriedenheit und Schmerzangabe von Patienten bei elektiven muskuloskeletalen Eingriffen vorhergesagt werden können, und ob diese Vorhersagepräzision abhängig ist vom ärztlichen Ausbildungsstand. Zudem sollte evaluiert werden, ob bei einem hoch standardisierten Eingriff, wie dem endoprothetischen Gelenkersatz, eine höhere Präzision möglich ist als im restlichen Operationsgut. Material und Methoden Die Ärzte der Klinik gaben präoperativ eine Einschätzung über die erwartete Zufriedenheit (Skala 0 – 10) und Schmerzstärke (NRS) der Patienten für den Zeitpunkt 6 Monate postoperativ ab. Zu diesem Zeitpunkt erfolgte dann die Erhebung dieser Parameter bei den Patienten. Die ärztliche Beurteilung erfolgte im Rahmen der präoperativen Visite am Vortag zur Operation. Unterschiedlich erfahrene Kollegen konnten so zeitgleich dieselben Patienten einschätzen. Verglichen wurden die Angaben von Ober- und Fachärzten mit denen der Assistenzärzte in Gegenüberstellung zu den Angaben der Patienten. Ergebnisse Insgesamt wurden 194 Einschätzungen ausgewertet. Durch die Operation konnte eine deutliche Schmerzbesserung erzielt werden (Median NRS prä-OP 6,25, post-OP 2,5, p Schlussfolgerung Auch wenn sich die ärztliche Einschätzung insgesamt mit den Patientenangaben annähernd deckt, so sind auf den Einzelfall bezogen starke Abweichungen zu verzeichnen, die auch in einem hoch standardisierten Kollektiv, wie dem der primären Endoprothetik, bestehen. Eine Zunahme der Einschätzungsqualität mit fortschreitendem ärztlichem Ausbildungsstand war nicht festzustellen. Hier lassen die Daten die Vermutung zu, dass über die technische Dimension hinaus andere Faktoren ebenfalls von entscheidender Bedeutung sind, wie z. B. die präoperative Erwartungshaltung des Patienten. Anspruch eines jeden Operateurs muss es daher sein, eine realistische Erwartung an die Operation mit dem Patienten gemeinsam zu entwickeln und sich dabei der eigenen Grenzen bez. der Vorhersagepräzision des Erfolgs bewusst zu sein.

Rhabdomyosarcomas (RMS) are soft tissue tumours treated with a combination of surgery and chemotherapy. However, mortality rates remain high in case of recurrences and metastatic disease due to drug resistance and failure to undergo apoptosis. Therefore, innovative approaches targeting specific signalling pathways are urgently needed. We analysed the impact of different hedgehog (Hh) pathway inhibitors on growth and survival of six RMS cell lines using MTS assay, colony formation assay, 3D spheroid cultures, flow cytometry and western blotting. Especially the glioma-associated oncogene family (GLI) inhibitor arsenic trioxide (ATO) effectively reduced viability as well as clonal growth and induced cell death in RMS cell lines of embryonal, alveolar and sclerosing, spindle cell subtype, whereas normal skeletal muscle cells were hardly compromised by ATO. Combination of ATO with itraconazole potentiated the reduction of colony formation and spheroid size. These results show that ATO is a promising substance for treatment of relapsed and refractory RMS by directly targeting GLI transcription factors. The combination with itraconazole or other chemotherapeutic drugs has the opportunity to enforce the treatment efficiency of resistant and recurrent RMS.

Objective The objective of this study was to investigate whether total knee arthroplasty (TKA) impairs the ability to perform an emergency stop. Design An automatic transmission brake simulator was developed to evaluate total brake response time. A prospective repeated-measures design was used. Forty patients (20 left/20 right) were measured 8 days and 6, 12, and 52 wks after surgery. Results Eight days postoperative total brake response time increased significantly by 30% in right TKA and insignificantly by 2% in left TKA. Brake force significantly decreased by 35% in right TKA and by 25% in left TKA during this period. Baseline values were reached at week 12 in right TKA; the impairment of outcome measures, however, was no longer significant at week 6 compared with preoperative values. Total brake response time and brake force in left TKA fell below baseline values at weeks 6 and 12. Brake force in left TKA was the only outcome measure significantly impaired 8 days postoperatively. Conclusion This study highlights that categorical statements cannot be provided. This study's findings on automatic transmission driving suggest that right TKA patients may resume driving 6 wks postoperatively. Fitness to drive in left TKA is not fully recovered 8 days postoperatively. If testing is not available, patients should refrain from driving until they return from rehabilitation.

The objective was to determine the role of dose intensive induction chemotherapy in patients with soft tissue sarcomas (STS) that were considered unresectable. Treatment consisted of 2–3 cycles of doxorubicin (Dox) and ifosfamide (Ifo) followed by high dose chemotherapy with ifosfamide, carboplatin, etoposide (HD-ICE) plus peripheral blood stem cell transplantation (PBSCT). 30 out of 631 consecutive patients, median age 46 years (21–62), with high grade STS were included. 29 patients completed at least 2 cycles of Dox/Ifo. HD-ICE was withheld because of progressive disease (PD) in 5 patients, neurotoxicity in 6 cases, insufficient peripheral blood stem cell (PBSC) mobilization, complete remission (CR) and refusal in 1 patient each. HD-ICE was associated with non-haematological grade III toxicity including emesis, mucositis, fever, neurotoxicity, and transaminase level elevation. Two additional patients attained a partial response after HD-ICE. Overall, 24 of 30 (80 %) patients underwent surgery, with complete tumor resections in 19 patients (63 % of all patients, 79 % of the operated subgroup); however, 2 of these required amputation. After a median follow up period of 50 months in surviving patients (range, 26–120), 5-year PFS and OS rates were 39 % and 48 %, respectively. Induction chemotherapy plus consolidation HD-ICE is generally feasible, but is associated with significant neurotoxicity. The advantage of HD-ICE over conventional dose chemotherapy plus external beam radiation therapy (EBRT) in non-resectable disease remains unproven.

As treatment results for high-risk soft tissue sarcoma are still disappointing, treatment intensification is warranted. We performed a retrospective analysis of multimodal preoperative treatment to evaluate the additional effect of concurrent chemotherapy and/or locoregional hyperthermia in comparison to radiotherapy alone. Between 1999 and 2011, 28 patients were treated with neoadjuvant radiotherapy to a median 45 Gy for high-risk soft tissue sarcoma. All tumors were deep-seated and grade 2 or 3, 86% (n = 24) larger than 5 cm. Multimodal treatment (n = 12) consisted of ifosfamide (n = 7), locoregional hyperthermia (n = 3), or both modalities (n = 2) concurrent to radiotherapy. Prognostic factors (grade, size, histology, location) were balanced in the groups with and without concurrent multimodal treatment. There was a significant improvement of disease-specific survival (100% vs. 70% at 3 years, p = 0.03) with multimodal treatment. Distant metastases-free survival was influenced, but was not statistically significant. Local control and disease-free survival did not differ in the two groups. Our data suggest that multimodal treatment with ifosfamide and/or locoregional hyperthermia in combination with neoadjuvant radiotherapy might improve outcome in high-risk soft tissue sarcomas.

Background aims Mesenchymal stromal/stem cells (MSCs) can be isolated from human bone marrow (BM), expanded ex vivo and identified via numerous surface antigens. Despite the importance of these cells in regenerative therapy programs, it is unclear whether the cell membrane signature defining MSC preparations ex vivo is determined during culture or may reflect an in vivo counterpart. BM-MSC phenotype in vivo requires further investigation. Methods To characterize cells in their natural BM environment, we performed multi-parametric immunohistochemistry on trabecular bone biopsy specimens from multiple donors and described cells by different morphology and micro-anatomic localization in relationship to a precise pattern of MSC antigen expression. Results Microscopically examined high-power field marrow sections revealed an overlapping in vivo expression of antigens characterizing ex vivo expanded BM-MSCs, including CD10, CD73, CD140b, CD146, GD2 and CD271. Expanding this panel to proteins associated with pluripotency, such as Oct4, Nanog and SSEA-4, we were able to identify different cellular populations in the human trabecular bone and BM expressing different progenitor cell markers. Conclusions Targeting several multipotency and pluripotency markers, we found that the BM contains identifiable and distinct progenitor cells further justifying their introduction for a wide range of applications in regenerative medicine.

SummaryThe LRRK2 mutation G2019S is the most common genetic cause of Parkinson’s disease (PD). To better understand the link between mutant LRRK2 and PD pathology, we derived induced pluripotent stem cells from PD patients harboring LRRK2 G2019S and then specifically corrected the mutant LRRK2 allele. We demonstrate that gene correction resulted in phenotypic rescue in differentiated neurons and uncovered expression changes associated with LRRK2 G2019S. We found that LRRK2 G2019S induced dysregulation of CPNE8, MAP7, UHRF2, ANXA1, and CADPS2. Knockdown experiments demonstrated that four of these genes contribute to dopaminergic neurodegeneration. LRRK2 G2019S induced increased extracellular-signal-regulated kinase 1/2 (ERK) phosphorylation. Transcriptional dysregulation of CADPS2, CPNE8, and UHRF2 was dependent on ERK activity. We show that multiple PD-associated phenotypes were ameliorated by inhibition of ERK. Therefore, our results provide mechanistic insight into the pathogenesis induced by mutant LRRK2 and pointers for the development of potential new therapeutics.

SummaryBackgroundAcetabular retroversion, excessive acetabular coverage and abnormal head-neck-junction with a so-called “pistol-grip-deformity” were added to the classical description of hip dysplasia to describe pathological hip morphology. The aim of the current study was the detection of pathological acetabular geometry in patients with an abnormal head-neck-junction.HypothesisFemoroacetabular impingement and hip dysplasia features are frequent in patients with end-stage osteoarthritis before 60 years of age.Materials and methodsWe analysed our data bank retrospectively for all patients who received a Total Hip Arthroplasty (THA) due to end-stage osteoarthritis before the age of 60 years. The pelvic-views and the Dunn-view of these patients were screened for an abnormal head-neck-junction by measuring the head-ratio and the alpha-angle. An orthopaedic surgeon and a radiologist did this independently. These radiographies were measured for signs of acetabular dysplasia, excessive acetabular coverage and crossing sign.ResultsA consecutive series of 135 total hip arthroplasties were performed in patients aged less or equal to 60 years because of end-stage osteoarthritis. From these, 81 patients were classified as having an abnormal head-neck-junction. The mean head-ratio in these 81 patients was 1.52±0.35, the mean alpha-angle was 62.5°±9.3°. The mean CE-angle of these 81 patients was 35.8°±10.4°, the mean CA-angle was 36.7°±5.7°, the mean depth-width ratio was 49.1±10, the mean extrusion index was 19.1±9.2 and the mean CCD-angle was 131.7°±7.3°. Of these 81hips, 14 had isolated pistol-grip-deformity, while 11 hips had associated dysplasia, 38 had excessive acetabular coverage, and 14 had crossing sign. In addition, a crossing sign was identified in four of the 11 dysplastic hips and 19 of the 38 of the hips having excessive acetabular coverage. There was no statistically significant difference in regard to the age between the four groups (P=0.087). In contrast, the hips that had excessive acetabular coverage had increased CE-angle (44.6°±7.2°) and decreased extrusion index (12.6±6.5) (P

Ewing sarcomas (ES) are rare mesenchymal tumours, most commonly diagnosed in children and adolescents. Arsenic trioxide (ATO) has been shown to efficiently and selectively target leukaemic blasts as well as solid tumour cells. Since multidrug resistance often occurs in recurrent and metastatic ES, we tested potential additive effects of ATO in combination with the cytostatic drugs etoposide and doxorubicin. The Ewing sarcoma cell lines A673, RD-ES and SK-N-MC as well as mesenchymal stem cells (MSC) for control were treated with ATO, etoposide and doxorubicin in single and combined application. Viability and proliferation (MTS assay, colony formation, 3D spheroid culture) as well as cell death induction (western blot analysis, flow cytometry) were analysed. In the MTS viability assays ATO treatment significantly reduced the metabolic activity of all three ES cell lines (A673, RD-ES and SK-N-MC) examined. Moreover, all ES cell lines were sensitive to etoposide, whereas MSC remained unaffected by the drug concentrations used. With the exception of ATO in RD-ES cells, all drugs induced apoptosis in the ES cell lines, indicated by caspase-3 and PARP cleavage. Combination of the agents potentiated the reduction of viability as well as the inhibitory effect on clonal growth. In addition, cell death induction was obviously enhanced in RD-ES and SK-N-MC cells by a combination of ATO and etoposide compared to single application. Summarised, the combination of low dose, physiologically easily tolerable ATO with commonly used etoposide and doxorubicin concentrations efficiently and selectively suppressed viability and colony formation in ES cell lines, whereas a combination of ATO and etoposide was favourable for cell death induction. In addition to an increase of the effectiveness of the cytostatic drugs and prevention of potential drug resistance, this approach may also reduce toxicity effects, since the individual doses can be reduced.

Stem cell therapies can promote neural repair and regeneration, yet controversy regarding optimal cell source and mechanism of action has slowed clinical translation, potentially due to undefined cellular heterogeneity. Single-cell resolution is needed to identify clinically relevant subpopulations with the highest therapeutic relevance. We combine single-cell microfluidic analysis with advanced computational modeling to study for the first time two common sources for cell-based therapies, human NSCs and MSCs. This methodology has the potential to logically inform cell source decisions for any clinical application.

Despite their paracrine activites, cardiomyogenic differentiation of bone marrow (BM)-derived mesenchymal stem cells (MSCs) is thought to contribute to cardiac regeneration. To systematically evaluate the role of differentiation in MSC-mediated cardiac regeneration, the cardiomyogenic differentiation potential of human MSCs (hMSCs) and murine MSCs (mMSCs) was investigated in vitro and in vivo by inducing cardiomyogenic and noncardiomyogenic differentiation. Untreated hMSCs showed upregulation of cardiac tropopin I, cardiac actin, and myosin light chain mRNA and protein, and treatment of hMSCs with various cardiomyogenic differentiation media led to an enhanced expression of cardiomyogenic genes and proteins; however, no functional cardiomyogenic differentiation of hMSCs was observed. Moreover, co-culturing of hMSCs with cardiomyocytes derived from murine pluripotent cells (mcP19) or with murine fetal cardiomyocytes (mfCMCs) did not result in functional cardiomyogenic differentiation of hMSCs. Despite direct contact to beating mfCMCs, hMSCs could be effectively differentiated into cells of only the adipogenic and osteogenic lineage. After intramyocardial transplantation into a mouse model of myocardial infarction, Sca-1(+) mMSCs migrated to the infarcted area and survived at least 14 days but showed inconsistent evidence of functional cardiomyogenic differentiation. Neither in vitro treatment nor intramyocardial transplantation of MSCs reliably generated MSC-derived cardiomyocytes, indicating that functional cardiomyogenic differentiation of BM-derived MSCs is a rare event and, therefore, may not be the main contributor to cardiac regeneration.

Disseminierte osteoblastische Knochenherde –Probleme bei der Diagnose der Osteopoikilie - Probleme bei der Diagnose der Osteopoikilie

IPACH I., MITTAG F., KOPP H.-G., KUNZE B., WOLF P. & KLUBA T. (2012) European Journal of Cancer Care21, 412–420 Clear-cell sarcoma of the soft tissue – a rare diagnosis with a fatal outcome Clear-cell sarcomas account for less than 1% of all soft tissue tumours. They most often occur in middle-aged adults as a deeply located lesion with predilection to the tendons and aponeuroses. The aim of the present study was to show possible influencing factors on the outcome after surgical treatment in a detailed case series. We reviewed the medical records of 11 patients with the diagnosis of a clear-cell sarcoma of the soft tissue. These cases were analysed with regard to age, gender, localisation, tumour size, recurrence free survival and overall survival. A minimum follow up of 12 months was achieved. The mean age at the point of diagnosis was 47.9 years. Metastases occurred after a mean of 19.2 months. In the cases with a tumour diameter >5 cm, metastases occurred earlier. When treated in a specialist centre, metastases occurred later. Patients died a mean of 18.4 months after developing metastatic disease. Patients with tumour size >5 cm at the point of primary diagnosis died earlier than patients with a tumour size

Early detection of pistol-grip-deformity may be beneficial in optimising the outcome and the cost effectiveness of surgery. It is important to identify reliable radiographic parameters in assessing femoro-acetabular impingement (FAI) to develop a treatment algorithm.Radiographs of 47 patients ranging from “pistol grip deformity” to a normal head-neck-junction were measured for alpha angle and head ratio, and then classified by two different observers. The Bland-Altman plot was used for inter- and intraobserver agreement of alpha angle and head ratio. Inter- and intraobserver agreement for classification of “pistol grip deformity” was determined using weighted Cohen's kappa coefficient.Observer I achieved a kappa coefficient of 0.97. Observer II achieved a kappa coefficient of 0.92. An interobserver kappa coefficient between 0.87 and 0.92 was achieved by using a classification system. By testing for interobserver agreement, a bias of –0.004 with an upper limit of 0.461 and a lower limit of –0.47 was seen for the ratio and a bias of –3.7 with an upper limit of 17.2 and a lower limit of –24.6 for the alpha angle.Therefore, poor results were seen for intra- and interobserver reliability by using only a single plane for classification of “pistol grip deformity”. The strength of agreement could be improved by using a classification system (based on two planes).

Incompatible blood group antigens are highly immunogenic and can cause graft rejections. Focusing on distinct carbohydrate- and protein-based membrane structures, defined by blood group antigens, we investigated human bone marrow-derived mesenchymal stem cells (MSCs) cultured in human serum. The presence of H (CD173), ABO, RhD, RhCE, RhAG, Kell, urea transporter type B (SLC14A1, previously known as JK), and Duffy antigen receptor of chemokines (DARC) was evaluated at the levels of genome, transcriptome and antigen. Fucosyltransferase-1 (FUT1), RHCE, KEL, SLC14A1 (JK) and DARC mRNA were transcribed in MSCs. FUT1 mRNA transcription was lost during differentiation. The mRNA transcription of SLC14A1 (JK) decreased during chondrogenic differentiation, while that of DARC increased during adipogenic differentiation. All MSCs synthesized SLC14A1 (JK) but no DARC protein. However, none of the protein antigens tested occurred on the surface, indicating a lack of associated protein function in the membrane. As A and B antigens are neither expressed nor adsorbed, concerns of ABO compatibility with human serum supplements during culture are alleviated. The H antigen expression by GD2dim+ MSCs identified two distinct MSC subpopulations and enabled their isolation. We hypothesize that GD2(dim+) H(+) MSCs retain a better 'stemness'. Because immunogenic blood group antigens are lacking, they cannot affect MSC engraftment in vivo, which is promising for clinical applications.

Purpose Two types of femoroacetabular impingement (FAI) are described as reasons for the early development of osteoarthritis of the hip. Cam impingement develops from contact between an abnormal head-neck junction and the acetabular rim. Pincer impingement is characterized by local or general overcoverage of the femoral head by the acetabular rim. Both forms might cause early osteoarthritis of the hip. A decreased head/neck offset has been recognized on AP pelvic views and labeled as "pistol grip deformity". The aim of the study was to develop a classification for this deformity with regard to the stage of osteoarthritis of the hip. Materials and methods 76 pelvic and axial views were analyzed for alpha angle and head ratio. 22 of them had a normal shape in the head-neck region and no osteoarthritis signs, 27 had a "pistol grip deformity" and osteoarthritis I and 27 had a "pistol grip deformity" and osteoarthritis II°-IV°. The CART method was used to develop a classification. Results There was a statistically significant correlation between alpha angle and head ratio. A statistically significant difference in alpha angle and head ratio was seen between the three groups. Using the CART method, we developed a three-step classification system for the "pistol grip deformity" with very high accuracy. This deformity was aggravated by increasing age. Conclusion Using this model it is possible to differentiate between normal shapes of the head-neck junction and different severities of the pistol grip deformity.

Brucella spp. are a main contributor to morbidity in endemic countries, especially in the Near and Middle East and the Mediterranean. However, in most western industrialised countries, cases are rare. Here, we report the case of a previously unrecognised chronic brucellosis with subsequent laboratory-acquired infection. A 64-year-old female presented to the orthopaedic department with loosening of her knee prosthesis after total knee arthroplasty five years earlier. Bacterial cultures were obtained in a preoperative puncture of the affected joint. About two weeks after replacement of the prosthesis bacterial growth was detected by chance in the preoperative culture and Brucella melitensis was diagnosed, whereas the intraoperative swab was negative. Serology revealed a chronic brucellosis while blood cultures were negative. Antibiotic treatment with rifampin and doxycycline was initiated. Two months later, a 32-year-old male physician who was involved in identifying the bacterial pathogen began to suffer from night sweats, weight loss and elevated temperatures. Another 4 weeks later, he developed hip pain and synovialitis. At that time, blood cultures and urine specimens were positive for Brucella melitensis. The serological results were consistent with acute brucellosis. In many European and North American countries, especially Germany, Brucella spp. are rarely isolated from clinical microbiological specimens and present a considerable risk for laboratory-acquired infections.

Osteosarcoma is one of the most common primary malignant bone tumours in childhood, mainly affecting the metaphysis of long extremity bones. In rare cases, patients present at time of diagnosis with multiple bone lesions, sometimes in the absence of pulmonary metastases. The pathology pattern of these multifocal osteosarcomas occurring with a rare incidence of 0.5-4% is not yet clear, and in spite of investigations in diagnosis and therapy, the prognosis is still poor. We report two cases of multifocal synchronous osteosarcoma. The age of both children at the time of tumour detection was 14 years. A synchronous or metachronous occurrence of multiple bone lesions, initially in the absence of pulmonary metastases was seen. In both cases, treatment consisted of neoadjuvant chemotherapy, oncologic surgery and adjuvant chemotherapy. Tumour response to chemotherapy was good in one patient, and poor in the other case. In both patients initial R0-resection of the tumours was performed. The disease-free time was 1 year before detection of pulmonary metastases or relapse. By the combination of chemotherapy and aggressive surgery the prognosis in multifocal osteosarcoma has been improved over the last years. Nevertheless, the survival time is still short and seems to be correlated with the initial histological tumour response to chemotherapy.

Ziel: Die monozentrische Studie vergleicht prospektiv die Beurteilbarkeit der Lage von Pedikelschrauben der LWS in der Multidetektor-CT mit einem 3-D-Bildwandler. Der Einfluss des Verfahrens sowie des Ausbildungsstands der Untersucher auf die Reproduzierbarkeit der Befunde wurden untersucht. Material und Methoden: Da die Untersuchung von Patienten vergleichend mit beiden Methoden durch die zustandige Ethikkommission abgelehnt wurde, mussten 2 Gruppen gebildet werden. Bei 23 Patienten mit 102 Pedikelschrauben wurden mit einem Arcadis-Orbic-3-D-Bildwandler intraoperativ untersucht und unabhangig voneinander von 3 Untersuchern befundet. Weitere 100 identische Schrauben wurden mit einem Multidetektor-CT postoperativ bei weiteren 16 Patienten uberpruft und wiederum in Bezug auf die korrekte Implantation hin von den 3 Untersuchern uberpruft. Der Kappa-Wert wurde zur Einschatzung der Reproduzierbarkeit der Auswertungen fur beide Techniken bestimmt. Daruber hinaus werteten die 3 Untersucher 24 Schrauben wiederholt aus und der Einfluss der Erfahrung auf die Haufigkeit des Auftretens von abweichenden Befunden wurde bestimmt. Ergebnisse: Die Befundung der CT-Untersuchungen wichen weniger voneinander ab im Vergleich zu denen mit dem Arcadis-Orbic-3-D. Die Ubereinstimmung aller 3 Untersucher betrug hierbei 79,4 vs. 65,1 %. Die entsprechenden Kappa-Werte betrugen 0,56 vs. 0,29. Besonders schlecht fiel die Ubereinstimmung fur die mediale Pedikelwand aus (70,0 vs. 50,0 %). Ein Einfluss des Ausbildungsstands lies sich statistisch nicht nachweisen. Schlussfolgerung: Die subjektiv vergleichend als schlechter empfundene Bildqualitat der ISO-C3D-Rekonstruktionen spiegelt sich in einer nachweisbar hoheren Abweichung der Befundung zwischen verschiedenen Untersuchern wider. Die Wertigkeit der Multidetektor-CT-Untersuchung bei der Beurteilung von lumbal implantierten Pedikelschrauben ist als uberlegen einzuschatzen. Purpose: This study was conducted to evaluate the reproducibility of the reading of lumbar pedicle screw scans using a C-arm-based imaging system in comparison to computed tomography. The influence of the technique and the experience of the rater should be determined. Materials and Methods: The lumbar spines of 23 patients were stabilized using 102 pedicle screws. The position of the screws was controlled intraoperatively using an Arcadis Orbic 3D scanner. All scans were evaluated independently by three raters. The position of the implants in reference to the pedicle walls was described. Additionally, another 100 lumbar pedicle screws in 16 patients were evaluated postoperatively with a multirow CT. Kappa according to Fleiss was calculated for the reproducibility of the rater statements. Each rater repeated the analysis of 24 screws to assess the intraobserver variance. Results: The reports of the CT scans showed significantly less variation. The consent of all 3 raters was achieved in 79.4 vs. 65.1 % of cases. The Kappa values were 0.56 and 0.29, respectively. Poor results were obtained especially for the medial pedicle wall (consent 70.0 vs. 50.0 %). The influence of the experience of the rater was not able to be verified. Conclusion: The image quality of the ISO C 3D is worse than that of multirow CT scans for the evaluation of lumbar pedicle screws. This causes greater variance among the rater reports. We stopped using the ISO C 3D technique intraoperatively for the implantation of lumbar pedicle screws.

Background aims Labeling of stem cells is crucial to allow tracking of stem cell homing and engraftment after transplantation. In this study we evaluated the influence of cell labeling procedures using clinically approved small particles of iron oxide (SPIO) with or without transfection reagents (TA) on functional parameters of human mesenchymal stem cells (MSC). Methods The study was approved by the institutional review board of the University of Tubingen, Germany. Seven populations of bone marrow (BM)-derived human mesenchymal stem cells (MSC) were labeled with SPIO alone or in combination with various TA. Directly after labeling and two passages after labeling migration assays, quantification of colony-forming units and quantitative evaluation of the differentiation potential were performed. Quantification of the cellular total iron load (TIL), determination of the cellular viability and electron microscopy were also performed. Results Labeling of mesenchymal stem cells with SPIO with or without TA did not affect cell viability and differentiation potential significantly. SPIO in combination with TA coated the cellular surface directly after labeling but was incorporated into the cells after two passages. Labeling of mesenchymal stem cells with TA led to a significant decrease of migration capacity. This effect was abolished after two passages. Labeling with and without TA led to a significant decrease in colony formation ability. This effect could also be observed after two passages. Conclusions The observed decrease of migration capacity and colony-formation ability was not associated with either TIL or localization of particles of iron oxide. SPIO labeling with and without TA had functional effects on human mesenchymal stem cells by decreasing the migration capacity and colony-formation ability of the stem cells.

PURPOSE Evaluation of low-dose full spine radiographs using fluoroscopic images for the assessment of the Cobb angle measurement in patients with scoliosis. MATERIAL AND METHODS Twenty-one consecutive patients (aged 10 - 27 years, mean age 14 years) with a conventional full spine examination (film speed class 800) underwent a follow-up exam using digital pulsed fluoroscopy (Multi Diagnost 4, Philips Medical Systems, Eindhoven, The Netherlands). The mean follow-up was 9 months. During a synchronized scan with a C-arm speed of 4 cm/sec fluoroscopic images were stored with a pulsed frequency of 3 images per second. The single images were merged and reconstructed to one image with the software Easy Spine (Philips medical Systems, Eindhoven, The Netherlands). The corresponding dose-area product values (DAP) of both methods were compared. Three independent observers assessed Cobb angles and image quality for each technique. RESULTS The mean DAP values for conventional imaging was 94.9 cGy x cm (2) and for fluoroscopy 7.8 cGy x cm (2), respectively. A significant dose reduction of 91.8 % (CI 91 % to 95 %) was calculated. The average absolute angle difference between the observers was found to be 2.7 degrees for conventional imaging and 2.4 degrees for the fluoroscopic method. Interobserver standard deviation of 2.9 degrees was lower than the 5.3 degrees for conventional images. Image quality was better in the conventional images. CONCLUSION Using the scanning method, we could achieve a mean reduction of the radiation dose of 92 %, while the accuracy of the Cobb angle measurements was comparable for both techniques despite of reduced image quality of digital fluoroscopy.

AIM A prospective analysis was performed to evaluate the relation between sagittal spinal morphology, gravity line position and morphological parameters of the pelvis and lumbar spine. METHOD 25 patients with a mean follow-up of 23 (15-37) years after thoracic Harrington distraction spondylodesis were included. Standing lateral full spine radiographs were performed while patients were standing on a force plate. Pelvic angles according to Duval-Beaupere were measured. RESULTS Pelvic incidence varied from 30 degrees to 76 degrees (mean 52.1 degrees + 12.5 degrees ). Mean sacral slope was 36.4 degrees (+ 7.8 degrees ). The average thoracic kyphosis (23.6 degrees + 14.3 degrees ) and lumbar lordosis (36.4 degrees + 9.2 degrees ) were diminished. The gravity line position was on average 10.9 mm (+ 21.6mm) posterior to the center of the femoral heads. CONCLUSION Our method quantifies the relationship between the gravity line position and pelvic parameters according to Duval-Beaupere. The homogenous study group of patients with AIS after Harrington spondylodesis is characterized by decreased lumbar and thoracic profile and posterior displacement of the gravity line position.

We studied prospectively the clinical and radiological course of 36 patients with localised spinal instability treated with vertebrectomy. A distractible titanium cage filled with polymethylmethacrylate(PMMA) in combination with transpedicular fixation was used to restore spinal stability. Mean follow-up was 16 (8-55) months. Average correction of the segmental kyphosis was 15.2 degrees (-2-29 degrees). During follow-up, a loss of correction of 0.9 degrees (-2-15 degrees) was seen. Segmental height was increased by 7.6 (0-14) mm on average with a mean loss of 1.6 (0-2) mm. Twenty-one patients without pre-operative neurological deficits were ambulatory without orthosis at discharge. Patients presenting with neurological deficits were unchanged (n=3) or improved (n=12).

Rhabdomyosarcomas (RMS) are the most prevalent soft tissue sarcomas affecting children and adolescents. Despite intensive treatment consisting of multimodal chemotherapy and surgery RMS patients diagnosed with metastatic disease expect long term survival rates of only 20%. Often multidrug resistance arises upon initial response emphasizing the need for new therapeutic drugs to improve treatment efficiency. Previously, we demonstrated the efficacy of the FDA approved drug arsenic trioxide (ATO) specifically inhibiting viability and clonal growth as well as inducing cell death in human RMS cell lines of different subtypes. In this study, we combined low dose ATO with lithium chloride (LiCl), which is approved as mood stabilizer for the treatment of bipolar disorder, but also inhibits growth and survival of different cancer cell types in pre-clinical research. Indeed, we could show additive effects of LiCl and ATO on viability reduction, decrease of colony formation as well as cell death induction. In the course of this, LiCl induced inhibitory glycogen synthase kinase-3β (GSK-3β) serine 9 phosphorylation, whereas glioma associated oncogene family 1 (GLI1) protein expression was particularly reduced by combined ATO and LiCl treatment in RD and RH-30 cell lines, showing high rates of apoptotic cell death. These results imply that combination of ATO with LiCl or another drug targeting GSK-3 is a promising strategy to enforce the treatment efficiency in resistant and recurrent RMS.

Myositis proliferans is a soft tissue neoplasia with rare incidence. In the most cases, it is localized in the region of the neck, shoulder, pelvis and thigh. Due to its rapid growth and histological picture, the tumour may appear as a malignant neoplasia. We report the case of a 29-year-old woman suffering from an increasing painful swelling of the left proximal lower leg. Performed biopsies and histological examinations provided the diagnosis of myositis proliferans adjacent to the fibula, which responded to local resection and did not recur after 2 years. We show the importance of adequate diagnostic and therapeutic approach to avoid unnecessary and probably radical overtherapy of the patient.

The genetic basis of many optic neuropathies remains unclear. Bonifert et al. show that deep intronic OPA1 mutations can account for the disease in a number of previously unsolved cases. Moreover, an OPA1 modifier variant can generate syndromic 'optic atrophy plus' phenotypes if combined in trans with a loss-of-function OPA1 mutation.The genetic diagnosis in inherited optic neuropathies often remains challenging, and the emergence of complex neurological phenotypes that involve optic neuropathy is puzzling. Here we unravel two novel principles of genetic mechanisms in optic neuropathies: deep intronic OPA1 mutations, which explain the disease in several so far unsolved cases; and an intralocus OPA1 modifier, which explains the emergence of syndromic 'optic atrophy plus' phenotypes in several families. First, we unravelled a deep intronic mutation 364 base pairs 3' of exon 4b in OPA1 by in-depth investigation of a family with severe optic atrophy plus syndrome in which conventional OPA1 diagnostics including gene dosage analyses were normal. The mutation creates a new splice acceptor site resulting in aberrant OPA1 transcripts with retained intronic sequence and subsequent translational frameshift as shown by complementary DNA analysis. In patient fibroblasts we demonstrate nonsense mediated messenger RNA decay, reduced levels of OPA1 protein, and impairment of mitochondrial dynamics. Subsequent site-specific screening of > 360 subjects with unexplained inherited optic neuropathy revealed three additional families carrying this deep intronic mutation and a base exchange four nucleotides upstream, respectively, thus confirming the clinical significance of this mutational mechanism. Second, in all severely affected patients of the index family, the deep intronic mutation occurred in compound heterozygous state with an exonic OPA1 missense variant (p.I382M; NM_015560.2). The variant alone did not cause a phenotype, even in homozygous state indicating that this long debated OPA1 variant is not pathogenic per se, but acts as a phenotypic modifier if it encounters in trans with an OPA1 mutation. Subsequent screening of whole exomes from > 600 index patients identified a second family with severe optic atrophy plus syndrome due to compound heterozygous p.I382M, thus confirming this mechanism. In summary, we provide genetic and functional evidence that deep intronic mutations in OPA1 can cause optic atrophy and explain disease in a substantial share of families with unsolved inherited optic neuropathies. Moreover, we show that an OPA1 modifier variant explains the emergence of optic atrophy plus phenotypes if combined in trans with another OPA1 mutation. Both mutational mechanisms identified in this study-deep intronic mutations and intragenic modifiers-might represent more generalizable mechanisms that could be found also in a wide range of other neurodegenerative and optic neuropathy diseases

A 67-year-old male patient developed progressive renal failure following successful treatment of a soft tissue sarcoma that comprised surgical resection after neoadjuvant radiochemotherapy with the application of doxorubicin (cumulative dose 180 mg/m²) and ifosfamide (cumulative dose 33 g/m²).Plasma creatinine concentration was elevated to 4.5 mg/dl. Upon detection of glucosuria and α1-microglobulinuria renal biopsy was performed.Histologic analysis revealed massively injured tubules that could be explained by a toxic effect of ifosfamide. Glomeruli were not affected and appeared normal. After two months of conservative therapy, the patient developed an uremic syndrome requiring hemodialysis. Ever since kidney function did not recover albeit preserved diuresis.Ifosfamide can cause end-stage renal disease by a tubulotoxic effect that may be the result of a selective intracellular uptake into the proximal tubule via the human organic cation transporter 2 (OCT2).

Introduction: Mesenchymal stromal cells (MSC) are a promising therapy for wound healing and regeneration of inflamed tissues. They are used clinically for different symptoms and diseases and are being investigated in clinical trials world wide at an increasing rate. However, depending on the application protocol and site of treatment, MSC may face an inflammatory environment. Objective: Nitric oxide (NO) is one of the soluble factors produced in acute and chronic inflammation and influences growth, apoptosis, proliferation and differentiation of cells. NO therefore my have an influence on MSC injected into inflamed sites. Thus we investigated the effects of NO radicals on human MSC. Methods: Human MSC were expanded and characterized. Expression of the mesenchymal linage markers was determined by flow cytometry and their tri-lineage differentiation was explored in vitro. MSC were incubated with the NO-donor sodium nitroprusside (SNP) at different concentrations (5 μM-5 mM) and over different periods of time (15 min–24 hrs), and analyzed for their respiratory activity, gene expression responses, cell signalling pathways, and differentiation potential. Results: Human MSC expressed the mesenchymal marker proteins CD73, CD90, CD105, CD146, but failed to express the hematopoietic markers CD11b, CD14, CD34, and CD45. Activation of the MSC in vitro by nitric oxide activated c-Raf-, p-38-MAPK, and p-JNK- mediated signalling in a dose dependent manner, and also significantly regulated genes involved in cellular proliferation (cyclin D1, GAS1), apoptotis (p53), and induced an intense nuclear factor E2-related factor (NRF2)-associated stress response. Moreover, NO inhibited the entry of MSC in the osteogenic differentiation pathway and NO-treated MSC expressed less of the transcription factor Runx2. In contrast, the expression of the adipogenic marker gene PPARγ2 remained unchanged. Conclusion: We conclude that NO modulates the metabolism of MSC and compromises their osteogenic differentiation potential, which may have detrimental consequences for bone remodelling or bone regeneration.

To show the possible effect of left- and right-side total hip arthroplasty (THA) on the ability to perform an emergency stop when driving a car.Inception cohort.A driving simulator using an actual car cabin, specifically developed for the experiment, was used for testing driving ability.Patients (N=40; 20 left-side THA/20 right-side THA) were tested preoperatively and in increments of 8 days and 6, 12, and 52 weeks after surgery.Left- and right-side THA.Reaction time, movement time, total brake response time (TBRT), and maximum brake force.Eight days postoperatively, measurements on driving performance indicated a slight worsening for all outcome parameters in patients after left-side THA and considerably more worsening in patients after right-side THA. For both patient groups, significant improvements in outcome measures were noted during the 1-year follow-up. Brake force declined significantly in patients with left-side THA (P=.012) and in patients after right-side THA (P.001). A total of 35% of the patients with right-side THA and 15% with left-side THA could not meet the 600 ms TBRT threshold 6 weeks postoperatively.Most patients who underwent right-side THA reached their preoperative baseline 6 weeks after surgery. Most of the patients with left-side THA showed no TBRT limitations 8 days postoperatively. Because of the patients' highly individual rehabilitation course and considering the possible consequences of the premature resumption of driving a motor vehicle, individual examination and recommendation are necessary.

Background Advising patients about when they can drive after surgery is common practice after arthroplasty of the knee or hip. In the literature, the preoperative braking performance values of the patients are frequently taken as the “safe” landmark. We hypothesised that osteoarthritis (OA), the most frequent reason for arthroplasty, already compromises the ability to perform an emergency stop. We expected that both Reaction Time (RT) and Movement Time (MT) as components of the Total Brake Response Time (TBRT), would be prolonged in patients with OA of the knee or hip in comparison with healthy subjects. We also expected maximum pressure levels on the brake pedal to be reduced in such cases. Methods A real car cabin was equipped with pressure sensors on the accelerator and brake pedals to measure RT, MT, TBRT and maximum Brake Force (BF) under realistic spatial constraints. Patients with OA of the knee (right n = 18, left n = 15) or hip (right n = 20, left n = 19) were compared with a healthy control group (n = 21). Results All measured values for TBRT in the control group remained below 600 ms. OA of the right hip or knee significantly prolonged the braking performance (right hip: TBRT p = 0.025, right knee: TBRT p

Introduction V.A.C. ® GranuFoam™ therapy is regularly used in the surgical therapy of infected wounds and soft tissue injuries. Silver nanoparticles can destroy bacterial cell walls and inhibit enzymes for cell replication. Silver dressings are therefore successfully used for many indications in wound therapy. In this study, we investigated the antimicrobial potency of ionic silver released from the silver-coated V.A.C. ® GranuFoam™ during vacuum therapy. Silver dressing was exposed to agar plates populated with bacteria to measure silver release. Materials and methods A total of 15 agar plates colonised with either Staphylococcus aureus populations or with Staphylococcus epidermidis , were loaded with V.A.C. GranuFoam Silver ® Dressing polyurethane foam (KCI, San Antonio, Texas). Each of 13 pieces of silver-coated foam was applied to an agar plate. Two plates were loaded with conventional black foam without any coating. After connecting to a vacuum pump, the vacuum therapy of the 15 plates lasted 5 days. The zone of inhibition of bacterial growth around the foam was measured daily. Silver release was also determined as a function of time. Results At each time point, there was evidence of silver in the agar independent of bacterial colonisation. The S. aureus agar showed a consecutive increase in silver concentration from baseline upon 48h after exposure to the negative pressure of V.A.C. therapy. An increasing mean silver level after 48, 72 and 96h was measured under V.A.C. therapy with a peak value after 120h. In contrast, the results from the S. epidermidis plates did not follow a linear pattern. At the beginning of vacuum therapy, we documented a rise in silver concentration. After 48–96h, the silver levels fluctuated. A maximum zone of inhibition in both bacterial colonised plates ( S. aureus and S. epidermidis ) was found 39h after the start of the V.A.C. GranuFoam Silver ® therapy. Conclusion From our results, we confirmed the antimicrobial effect of the silver ions against S. aureus and S. epidermidis under continuous V.A.C. GranuFoam ® Silver therapy with a negative pressure of 25mmHg. Furthermore we could quantify the amounts of silver, which were released from the foam under negative pressure as a function of time.

Advanced adult soft-tissue sarcomas (STSs) are rare tumors with a dismal prognosis and limited systemic treatment options. STSs may originate from mesenchymal stem cells (MSCs); the latter have mainly been isolated from adult bone marrow as plastic-adherent cells with differentiation capacity into mesenchymal tissues. Recently, a panel of antibodies has been established that allows for the prospective isolation of primary MSCs with high selectivity. Similar to cancer stem cells in other malignancies, sarcoma stem cells may bear immunophenotypic similarity with the corresponding precursor, that is, MSCs. We therefore set out to establish the expression pattern of MSC markers in sarcoma cell lines and primary tumor samples by flow cytometry. In addition, fibroblasts from different sources were examined. The results document a significant amount of MSC markers shared by sarcoma cells. The expression pattern includes uniformly expressed markers, as well as MSC markers that only stained subpopulations of sarcoma cells. Expression of W5C5, W8B2 (tissue nonspecific alkaline phosphatase [TNAP]), CD344 (frizzled-4), and CD271 marked subpopulations displaying increased proliferation potential. Moreover, CD271+ cells displayed in vitro doxorubicin resistance and an increased capacity to form spheres under serum-free conditions. Interestingly, another set of antigens, including the bona fide progenitor cell markers CD117 and CD133, were not expressed. Comparative expression patterns of novel MSC markers in sarcoma cells, as well as fibroblasts and MSCs, are presented. Our data suggest a hierarchical cytoarchitecture of the most common adult type sarcomas and introduce W5C5, TNAP, CD344, and CD271 as potential sarcoma progenitor cell markers.

Background Mesenchymal stromal cells (MSCs) are attractive for cell-based therapies ranging from regenerative medicine and tissue engineering to immunomodulation. However, clinical efficacy is variable and it is unclear how the phenotypes defining bone marrow (BM)-derived MSCs as well as donor characteristics affect their functional properties. Methods BM-MSCs were isolated from 53 (25 female, 28 male; age: 13 to 80 years) donors and analyzed by: (1) phenotype using flow cytometry and cell size measurement; (2) in vitro growth kinetics using population doubling time; (3) colony formation capacity and telomerase activity; and (4) function by in vitro differentiation capacity, suppression of T cell proliferation, cytokines and trophic factors secretion, and hormone and growth factor receptor expression. Additionally, expression of Oct4, Nanog, Prdm14 and SOX2 mRNA was compared to pluripotent stem cells. Results BM-MSCs from younger donors showed increased expression of MCAM, VCAM-1, ALCAM, PDGFRβ, PDL-1, Thy1 and CD71, and led to lower IL-6 production when co-cultured with activated T cells. Female BM-MSCs showed increased expression of IFN-γR1 and IL-6β, and were more potent in T cell proliferation suppression. High-clonogenic BM-MSCs were smaller, divided more rapidly and were more frequent in BM-MSC preparations from younger female donors. CD10, β1integrin, HCAM, CD71, VCAM-1, IFN-γR1, MCAM, ALCAM, LNGFR and HLA ABC were correlated to BM-MSC preparations with high clonogenic potential and expression of IFN-γR1, MCAM and HLA ABC was associated with rapid growth of BM-MSCs. The mesodermal differentiation capacity of BM-MSCs was unaffected by donor age or gender but was affected by phenotype (CD10, IFN-γR1, GD2). BM-MSCs from female and male donors expressed androgen receptor and FGFR3, and secreted VEGF-A, HGF, LIF, Angiopoietin-1, basic fibroblast growth factor (bFGF) and NGFB. HGF secretion correlated negatively to the expression of CD71, CD140b and Galectin 1. The expression of Oct4, Nanog and Prdm14 mRNA in BM-MSCs was much lower compared to pluripotent stem cells and was not related to donor age or gender. Prdm14 mRNA expression correlated positively to the clonogenic potential of BM-MSCs. Conclusions By identifying donor-related effects and assigning phenotypes of BM-MSC preparations to functional properties, we provide useful tools for assay development and production for clinical applications of BM-MSC preparations.

Mesenchymal stromal cells (MSC) are differentiation competent cells and may generate, among others, mature osteoblasts or chondrocytes in vitro and in vivo. Laminin-5 and type I collagen are important components of the extracellular matrix. They are involved in a variety of cellular and extracellular activities including cell attachment and osteogenic differentiation of MSC. MSC were isolated and expanded using media conforming good medical practice (GMP)-regulations for medical products. Cells were characterized according to the defined minimal criteria for multipotent MSC. MTT- and BrdU-assays were performed to evaluate protein-dependent (laminin-5, laminin-1, type I collagen) metabolic activity and proliferation of MSC. MSC-attachment assays were performed using protein-coated culture plates. Osteogenic differentiation of MSC was measured by protein-dependant mineralization and expression of osteogenic marker genes (osteopontin, alkaline phophatase, Runx2) after three, seven and 28 days of differentiation. Marker genes were identified using quantitative reverse-transcription polymerase chain reaction. Expansion of MSC in GMP-conforming media yielded vital cells meeting all minimal criteria for MSC. Attachment assay revealed a favorable binding of MSC to laminin-5 and type I collagen at a protein concentration of 1-5 fmol/mL. Compared to plastic, osteogenic differentiation was significantly increased by laminin-5 after 28 days of culture (P

Local adjuvant treatment of giant cell tumours (GCTs) of the bone with phenol has led to a significant reduction in recurrence rates. In the current study, the optimal phenol concentration and duration of intralesional exposure were evaluated. Specimens of GCTs were exposed to various concentrations of phenol solution (6, 60 and 80%) for either 1 or 3 min. Following embedding in glutaraldehyde, the tumour cell layers were examined by transmission electron microscopy. Destroyed cell organelles indicated the penetration depth as a sign of denaturation. Incubation of GCT specimens with 6% phenol solution for 3 min resulted in the most tissue damage and the deepest tissue penetration of ∼200 μm. Incubation with 60 and 80% phenol solution reached a penetration depth of only ∼100 μm. Phenol instillation may be used for the treatment of small scattered cellular debris following intralesional curettage; however, it is not suitable for treatment of remaining solid tumour tissue of GCT. The use of high phenol concentrations has no benefit and increases the risk of local or systemic intoxication.

Osteogenesis imperfecta (OI) is characterized by different signs including increased bone fragility, short stature, blue sclera, abnormal tooth growth and often secondary immobility. No curative therapy has been found for this rare disease up to now, and different pharmacological substances have been tried as treatment for severe forms of OI. Promising results were seen with intravenous bisphosphonates in the treatment of patients with OI. The aim of present study was to show the effect of intravenous ibandronate therapy on bone density and bone metabolism markers. We analyzed the data of 27 patients with the diagnosis of OI who were treated off-label with intravenous ibandronate. Ibandronate was administered by intravenous infusion every three months at a dosage of 0.3-2 mg. Bone turnover markers and bone density were measured before starting therapy and every three months during treatment. Bone density was measured by using an ultrasound imaging system providing an accurate image of the calcaneus and by evaluating broadband ultrasound attenuation (BUA). Twenty-seven patients were treated with intravenous iban- dronate during the observation period. 18 were female. The mean age of all patients was 23.9 years ± 19.6 (range 4-63). Seventeen patients were categorized to have OI Type I, 5 patients to have OI Type III and 5 patients to have OI Type IV. There was a statistically significant decrease in total alkaline phosphatase (P