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Background Aseptic implant loosening and periprosthetic bone loss are major problems after total hip arthroplasty (THA). We present an in vivo method of computed tomography (CT) assisted osteodensitometry after THA that differentiates between cortical and cancellous bone density (BD) and area around the femoral component. Method Cortical and cancellous periprosthetic femoral BD (mg CaHA/mL), area (mm2) and contact area between the prothesis and cortical bone were determined prospectively in 31 patients 10 days, 1 year, and 6 years after uncemented THA (mean age at implantation: 55 years) using CT-osteodensitometry. Results 6 years postoperatively, cancellous BD had decreased by as much as 41% and cortical BD by up to 27% at the metaphyseal portion of the femur; this decrease was progressive between the 1-year and 6-year examinations. Mild cortical hypertrophy was observed along the entire length of the diaphysis. No statistically significant changes in cortical BD were observed along the diaphysis of the stem. Interpretation Periprosthetic CT-assisted osteodensitometry has the technical ability to discriminate between cortical and cancellous bone structures with respect to strain-adapted remodeling. Continuous loss of cortical and cancellous BD at the femoral metaphysis, a homeostatic cortical strain configuration, and mild cortical hypertrophy along the diaphysis suggest a diaphyseal fixation of the implanted stem. CT-assisted osteodensitometry has the potential to become an effective instrument for quality control in THA by means of in vivo determination of periprosthetic BD, which may be a causal factor in implant loosening after THA. [ABSTRACT FROM AUTHOR]

Hypothesis And/or Background: Extensive, irreparable rotator cuff tears remain a surgical challenge and multiple treatment options are proposed and currently in use. To biomechanically compare superior glenohumeral translation, subacromial contact pressures, and area in a box-shaped reconstruction using the long head of the biceps tendon (LHBT) in an irreparable supraspinatus tendon tear model., Methods: Seven cadaveric shoulders (mean age 61 years; range 32-84 years; standard deviation 22.3) were tested with a custom testing rig used to evaluate superior translation; subacromial contact pressures; and areas at 0°, 30°, and 60° of glenohumeral abduction. Conditions tested included the native state; a complete tear of the supraspinatus tendon; a wide box-shaped, double-bundle LHBT superior capsular reconstruction (BS-SCR); and a narrow BS-SCR., Results: Compared with the wide BS-SCR, the narrow BS-SCR had statistically significantly lower median contact pressure at 30° and 60°. The subacromial contact area showed a statistically significant difference at 0° (P = .001) and 30° (P = .004) for the narrow BS-SCR compared with the wide BS-SCR. At an abduction angle of 0°, the narrow BS-SCR could restore superior translation significantly better compared with the wide construct. For all angles, the wide and narrow BS-SCR increased the median subacromial distance statistically significantly. The contact areas in 30° and 60° of abduction were higher for all scenarios, both peaking in the intact state in 30° with approximately 600 mm 2 ., Conclusions: In comparison to a wide BS-SCR, a narrow BS-SCR using the LHBT has biomechanical advantages in regard to subacromial contact pressures, the subacromial contact areas, as well as the acromiohumeral distance. The width of the reconstruction, therefore, has a direct influence on the success of the technique., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Introduction: The aim of this study was to evaluate the range of motion (ROM), elbow function and predictors for good elbow function after conservative treatment of non-displaced radial head fractures., Material and Methods: All patients with non-displaced radial head fractures (displacement < 2 mm), that were diagnosed between January 1st 2017 and December 31st 2021 in a level I trauma center, were included in this retrospective case series and the charts were evaluated for ROM and elbow function. Elbow function was categorized as "good" or "bad" depending on the ROM measured defined by Morrey et al. Overall, 73 patients (33 male, 40 female) with an average age of 38 years (+/- 13 years) could be included., Results: Conservative treatment had good clinical results for ROM and elbow function. After 6 weeks mean flexion was 131° (SD 13°), extension 8° (SD 7°), Pronation 83° (SD 11°) and Supination 83° (SD 13). Patients with a good elbow function after one week showed a good elbow function after completing the treatment., Conclusions: A clinical assessment after one week should always be performed and the study showed that it is a good predictor for good elbow function. In cases of bad elbow function further controls should be considered., (© 2024. The Author(s).)

Introduction: Osteochondrosis dissecans (OCD) at the capitellum is a common pathology in young patients. Although arthroscopic interventions are commonly used, there is a lack of information about the accessibility of the defects during elbow arthroscopy by using standard portals., Materials and Methods: An elbow arthroscopy using the standard portals was performed in seven fresh frozen specimens. At the capitellum, the most posterior and anterior cartilage surface reachable was marked with K-wires. Using a newly described measuring method, we constructed a circular sector around the rotational center of the capitellum. The intersection of K-wire "A" and "B" with the circular sector was marked, and the angles between the K-wires and the Rogers line, alpha angle for K-Wire "A" and beta angle for K-wire "B", and the corridor not accessible during arthroscopy was digitally measured., Results: On average, we found an alpha angle of 53° and a beta angle of 104°. Leaving a sector of 51° which was not accessible via the standard portals during elbow arthroscopy., Conclusion: Non-accessible capitellar lesions during elbow arthroscopy should be considered preoperatively, and the informed consent discussion should always include the possibility of open procedures or the use of flexible instruments., (© 2024. The Author(s).)

Colorectal cancer (CRC) is a heterogeneous disease. More insight into the biological diversity of CRC is needed to improve therapeutic outcomes. Established CRC cell lines are frequently used and were shown to be representative models of the main subtypes of CRC at the genomic and transcriptomic level. In the present work, we established stable, luciferase expressing derivatives from 10 well-established CRC cell lines, generated spheroids and subcutaneous xenograft tumors in nude mice, and performed comparative characterization of these model systems. Transcriptomic analyses revealed the close relation of cell lines with their derived spheroids and xenograft tumors. The preclinical model systems clustered with patient tumor samples when compared to normal tissue thereby confirming that cell-line-based tumor models retain specific characteristics of primary tumors. Xenografts showed different differentiation patterns and bioluminescence imaging revealed metastatic spread to the lungs. In addition, the models were classified according to the CMS classification system, with further sub-classification according to the recently identified two intrinsic epithelial tumor cell states of CRC, iCMS2 and iCMS3. The combined data showed that regarding primary tumor characteristics, 3D-spheroid cultures resemble xenografts more closely than 2D-cultured cells do. Furthermore, we set up a bioluminescence-based spheroid cytotoxicity assay in order to be able to perform dose-response relationship studies in analogy to typical monolayer assays. Applying the established assay, we studied the efficacy of oxaliplatin. Seven of the ten used cell lines showed a significant reduction in the response to oxaliplatin in the 3D-spheroid model compared to the 2D-monolayer model. Therapy studies in selected xenograft models confirmed the response or lack of response to oxaliplatin treatment. Analyses of differentially expressed genes in these models identified CAV1 as a possible marker of oxaliplatin resistance. In conclusion, we established a combined 2D/3D, in vitro/in vivo model system representing the heterogeneity of CRC, which can be used in preclinical research applications.

Tumor-associated carbonic anhydrases IX (CAIX) and XII (CAXII) have long been in the spotlight as potential new targets for anti-cancer therapy. Recently, CAIX/CAXII specific inhibitor SLC-0111 has passed clinical phase I study and showed differential response among patients with colorectal cancer (CRC). CRC can be classified into four different consensus molecular subgroups (CMS) showing unique expression patterns and molecular traits. We questioned whether there is a CMS-related CAIX/CAXII expression pattern in CRC predicting response. As such, we analyzed transcriptomic data of tumor samples for CA9/CA12 expression using Cancertool. Protein expression pattern was examined in preclinical models comprising cell lines, spheroids and xenograft tumors representing the CMS groups. Impact of CAIX/CAXII knockdown and SLC-0111 treatment was investigated in 2D and 3D cell culture. The transcriptomic data revealed a characteristic CMS-related CA9/CA12 expression pattern with pronounced co-expression of both CAs as a typical feature of CMS3 tumors. Protein expression in spheroid- and xenograft tumor tissue clearly differed, ranging from close to none (CMS1) to strong CAIX/CAXII co-expression in CMS3 models (HT29, LS174T). Accordingly, response to SLC-0111 analyzed in the spheroid model ranged from no (CMS1) to clear (CMS3), with moderate in CMS2 and mixed in CMS4. Furthermore, SLC-0111 positively affected impact of single and combined chemotherapeutic treatment of CMS3 spheroids. In addition, combined CAIX/CAXII knockdown and more effective treatment with SLC-0111 reduced clonogenic survival of CMS3 modelling single cells. In conclusion, the preclinical data support the clinical approach of targeted CAIX/CAXII inhibition by showing linkage of expression with response and suggest that patients with CMS3-classified tumors would most benefit from such treatment.

Background Aims: The immunomodulating capacity of multipotent mesenchymal stromal cells (MSCs) qualifies them as a therapeutic tool in several diseases. However, repeated transplantation with products of reproducible characteristics may be required. This could be achieved with cryopreserved aliquots of Good Manufacturing Practice (GMP)-grade MSCs. However, the impact of cryopreservation on the characteristics of GMP-MSCs is ill defined., Methods: We produced fresh and cryopreserved MSCs from human donors with a xenogen-free GMP protocol. Immunogenicity and immunomodulating capacity were tested in co-culture with putative recipient-specific peripheral blood mononuclear cells (PBMCs). Risk of malignant transformation was assessed in vitro and in vivo., Results: Cryopreservation had no impact on viability and consensus criteria of MSCs. In co-culture with PBMCs, MSCs showed low immunogenicity and suppressed mitogen-stimulated proliferation of PBMC irrespective of cryopreservation. Cytogenetic aberrations were not observed consistently in fresh and cryopreserved products, and no signs of malignant transformation occurred in functional assays. MSC products from an elderly pretreated donor showed reduced functional quality, but imminent failure of functional criteria could be detected by an increased population doubling time in early passages., Discussion: This study is the first systematic analysis on cryopreservation of xenogen-free human bone marrow-derived GMP-MSCs. The data support that cryopreservation does not alter the characteristics of the cells and thus may allow the generation of products for serial transplantation. In addition, the protocol allowed early detection of MSC products with low functional capacity., (Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)

Purpose: Intramedullary nailing and locked plating for fixation of olecranon fractures has recently gained popularity. However, these two new technologies have not been compared for their biomechanical efficacy. The aim of this study was to evaluate the biomechanical stability of two newly designed fracture fixation devices for treating olecranon fractures during dynamic continuous loading: the ION intramedullary locking nail and the LCP precontoured locking compression plate., Methods: Simulated oblique olecranon fractures were created in eight pairs of fresh-frozen cadaver ulnae and stabilised using either the LCP or ION. Specimens were then subjected to continuous dynamic loading (from 25 to 200 N), with a continuous angle alteration between 0° and 90° of flexion, to perform a matched-pairs comparison. Significant differences in the distance between markers surrounding the fracture gap was determined using the Wilcoxon test after four and 300 loading cycles., Results: The ION resulted in significantly less displacement in the fracture gap at 0° extension (P = 0.036), 45° flexion (P = 0.035) and 90° flexion (P = 0.017) after 300 cycles of continuous loading. The measured displacements were small and were probably not of clinical significance. No mechanical failure or hardware migration was seen with either fixation technique., Conclusion: This study shows significantly less micromotion for the ION than for the LCP in treating oblique olecranon fractures after 300 cycles of dynamic loading. Both implant types could be appropriate surgical techniques for fixation of selected olecranon fractures and osteotomies.

Radial head fractures represent the most common elbow fractures. Undisplaced fractures usually occur in isolation and can be treated nonsurgically. Displaced fractures should be treated surgically. Simple two-part fractures can easily be handled by osteosynthesis, but comminuted fractures pose a major problem for reconstruction. As the radial head is an important stabilizer of the elbow joint-especially in the context of concomitant ligamentous injuries-its resection may lead to pain, limited range of motion, and instability. Therefore, radial head resection is not recommended for the acute situation and open reduction internal fixation (ORIF) or prosthetic replacement should be aimed for. Complications such as secondary loss of fixation, radial head necrosis, and nonunion due to insufficient stability of the osteosynthesis have often been described. Therefore, prosthetic replacement is recommended if stable reconstruction is impossible. With the development of new locking plates especially designed for the maintenance of radial head fractures, the indications for osteosynthesis may be extended. As radial head fractures are complicated by a high percentage of ligamentous injuries and concomitant elbow fractures such as the coronoid, capitellum, and proximal ulna, these additional injuries have to be taken into account. The current treatment concepts are discussed within this paper.

Background: The proximal radius features a complex anatomy. Several studies have been published on the anatomy using different technical approaches; however, most of these studies were conducted with a special focus on parameters relevant to radial prosthetic design. The purpose of our study was to explore the complex geometry of the proximal radius with regard to fracture implant design., Methods: Our computed tomography-based measurements of 78 multiplanar reformatted radii allow for exact assessment of its geometry and offer a scientific rationale towards the design of fracture implants. We conducted measurements on the radial head, the radial neck, the radial tuberosity, the radial head-to-neck angle, and the safe zone., Results: A wide range of normal anatomy has been demonstrated for all parameters. Sex differences are statistically significant in all registered parameters, except the radial head-to-neck angle. Although measurements of maximum vs minimum radial head, neck, and tuberosity diameters show close correlation, diameter-to-length correlations, such as radial head diameter vs radial head height and radial neck diameter vs radial neck length, are low., Conclusions: Besides the wide range in size, intraindividual parameter variations have to be taken into account in the design of anatomically precontoured plates. The results of this study indicate that these plates will still need to offer the ability of "bend to match.", (Copyright © 2012 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Mosby, Inc. All rights reserved.)

A continuous monitoring of the whole tumor burden of individuals in orthotopic tumor models is a desirable aim and requires non-invasive imaging methods. Here we investigated whether quantification of a xenograft tumor intrinsic fluorescence signal can be used to evaluate tumor growth and response to chemotherapy. Stably fluorescence protein (FP) expressing cell clones of colorectal carcinoma and germ cell tumor lines were generated by lentiviral transduction using the FPs eGFP, dsRed2, TurboFP635, and mPlum. Applying subcutaneous tumor models in different experimental designs, specific correlations between measured total fluorescence intensity (FI) and the tumor volume (V) could be established. The accuracy of correlation of FI and V varied depending on the cell model used. The application of deep-red FP expressing xenografts (TurboFP635, mPlum) was observed to result in improved correlations. This was also reflected by the results of a performed error analysis. In a model of visceral growing mPlum tumors, measurements of FI could be used to follow growth and response to chemotherapy. However, in some cases final necropsy revealed the existence of additional, deeper located tumors that had not been detected in vivo by their mPlum signal. Consistently, only the weights of the tumors that were detected in vivo based on their mPlum signal correlated with FI. In conclusion, as long as tumors are visualized by their fluorescence signal the FI can be used to evaluate tumor burden. Deep-red FPs are more suitable for in vivo applications as compared to eGFP and dsRed2.

Tumor-integrating multipotent mesenchymal stromal cells (MSC) expressing transgenes with anti-tumor activity may serve as vehicles for tumor therapy. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) represents such a factor; however, TRAIL-resistant tumor cells exist. Based on our previous work, here we investigated whether MSC with lentiviral TRAIL expression (TRAIL-MSC) inhibit the growth of TRAIL-resistant colorectal carcinoma (CRC) cells. Our data show that TRAIL-MSC induce apoptosis in selected TRAIL-resistant CRC cell lines and effectively inhibit the growth of TRAIL-resistant HCT8 cells. This sensitization to TRAIL-induced apoptosis required the presence of MSC-expressed TRAIL. However, for the first time we show that selected CRC cells are resistant to TRAIL-MSC. In the cell line HT29, this resistance could be overcome by concomitant subapoptotic genotoxic damage in vitro. However, such sensitization was not achieved in vivo as treatment of mixed HT29/TRAIL-MSC xenografts with 5-FU rather resulted in enhanced growth. Taken together, our data prove that TRAIL-MSC overcome TRAIL resistance in selected CRC cells through direct intercellular interaction and may, therefore, represent a clinical tool to overcome TRAIL resistance. However, such potential clinical use requires further preclinical studies as our data also prove that TRAIL-MSC-resistant CRC cells exist. Our data add to the notion that TRAIL resistance of CRC cells is conferred by different mechanisms.

We evaluated the feasibility and toxicity of bevacizumab in combination with sequential high-dose (HD) ifosfamide, carboplatin and etoposide refractory to standard chemotherapy in patients with sarcoma and germ cell cancer (GCC). Sixteen patients (13 sarcomas, 3 GCC) received SD-ICE followed by 4 cycles of HD-ICE, qd22 with stem cell support in combination with bevacizumab. All 16 patients were evaluable for toxicity and efficacy, and received 51 cycles (median 3.3). There was no increase in toxicity except of a relatively high incidence of ifosfamide encephalopathy in 17 cycles when compared with previous HD-ICE protocols. One almost complete response in the patient with GCC, previously progressive with three preceding protocols, was observed. Six patients had a partial response (sarcoma 4/13 patients; GCC 2/3 patients), and five patients stable disease (sarcoma 5/13 patients). The median PFS/OS for sarcoma was 5 months (confidence interval (CI): 3.1-6.9) and 13 months (CI: 3.6-24.4), respectively. To our knowledge, this is the first report of the addition of bevacizumab to HD-ICE. This combination did not show new unexpected toxicities except for a relatively high rate of ifosfamide encephalopathy. The efficacy in these heavily pretreated patients including possible reversal of chemotherapy resistance by the addition of bevacizumab indicates a possible potential of bevacizumab in this combination.

Owing to the lack of fresh human bones, formalin-fixed specimens are frequently used in biomechanical testing. However, formalin fixation is assumed to affect the biomechanical properties of bone. The aim of this study was to compare axial and torsional stiffness and bone mineral density in fresh and embalmed human bones. The subtrochanteric regions of 12 pairs of fresh human femora were randomised into two groups for paired comparison. After bone mineral density measurement, one group was preserved in 4% formalin. After 6 weeks, bone mineral density was remeasured and each specimen underwent axial and torsional loading. The formalin group showed significant higher stiffness values for torsional and axial loads than the fresh group. Axial stiffness increased by 14.1%, whereas torsional stiffness increased by 14.3%. These differences were not reflected in bone mineral density values. Formalin fixation significantly influences the stiffness of human cadaveric bones. Fresh bones represent the in vivo conditions better than formalin fixed bones.

Background: The purpose of this study was to compare the stability of a 2.4mm palmar locking compression plate and a new intramedullary nail-plate-hybrid Targon DR for dorsally comminuted distal radius fractures., Methods: An extraarticular 10mm dorsally open wedge osteotomy was created in 8 pairs of fresh frozen human radii to simulate an AO-A3-fracture. The fractures were stabilized using one of the fixation methods. The specimens were loaded axially with 200 N and dorsal-excentrically with 80 N. 2000cycles of dynamic loading and axial loading-to-failure were performed., Findings: Axial loading revealed that intramedullary osteosynthesis (Targon DR: 369 N/mm) was significantly (p=0.017) stiffer than plate osteosynthesis (Locking compression plate: 131 N/mm). With 214 N/mm the intramedullary nail also showed higher stability during dorsal excentric loading than the Locking compression plate with 51 N/mm (p=0.012). After 2000 cycles of axial loading with 80 N the Targon DR-group was significantly stiffer than the Locking compression plate-group under both loading patterns. Neither group showed significant changes in stiffness after 2000 cycles. Under dorsal excentric loading the Targon DR-group was still significantly stiffer with 212 N/mm than the Locking compression plate-group with 45 N/mm (p=0.012). The load to failure tests demonstrated higher stability of intramedullary nailing (625 N) when compared to plate osteosynthesis (403 N) (p<0.025)., Interpretation: The study shows that intramedullary fixation of a distal AO-A3 radial fracture is biomechanically more stable than volar fixed-angle plating under axial and dorsal-excentric loading in an experimental setup., (2010 Elsevier Ltd. All rights reserved.)

Colorectal carcinoma (CRC) constitutes a common malignancy with limited therapeutic options in metastasized stages. Mesenchymal stem cells (MSC) home to tumours and may therefore serve as a novel therapeutic tool for intratumoral delivery of antineoplastic factors. Tumour necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) which promises apoptosis induction preferentially in tumour cells represents such a factor. We generated TRAIL-MSC by transduction of human MSC with a third generation lentiviral vector system and analysed their characteristics and capacity to inhibit CRC growth. (1) TRAIL-MSC showed stable transgene expression with neither changes in the defining MSC characteristics nor signs of malignant transformation. (2) Upon direct in vitro coculture TRAIL-MSC induced apoptosis in TRAIL-sensitive CRC-cell lines (DLD-1 and HCT-15) but also in CRC-cell lines resistant to soluble TRAIL (HCT-8 and SW480). (3) In mixed subcutaneous (s.c.) xenografts TRAIL-MSC inhibited CRC-tumour growth presumably by apoptosis induction but a substantial proportion of TRAIL-MSC within the total tumour cell number was needed to yield such anti-tumour effect. (4) Systemic application of TRAIL-MSC had no effect on the growth of s.c. DLD-1 xenografts which appeared to be due to a pulmonary entrapment and low rate of tumour integration of TRAIL-MSC. Systemic TRAIL-MSC caused no toxicity in this model. (5) Wild-type MSC seemed to exert a tumour growth-supporting effect in mixed s.c. DLD-1 xenografts. These novel results support the idea that lentiviral TRAIL-transgenic human MSC may serve as vehicles for clinical tumour therapy but also highlight the need for further investigations to improve tumour integration of transgenic MSC and to clarify a potential tumour-supporting effect by MSC., (© 2009 The Authors Journal compilation © 2010 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd.)

The biological basis for manifestation of chemotherapy resistance in metastatic testicular germ cell tumors (GCT) remains obscure and is of particular clinical interest. In nonseminomatous GCT (NSGCT) the pluripotent embryonal carcinoma (EC) cells are the precursors of the manifold differentiated structures but also drive the malignant growth. They are known to be hypersensitive towards DNA-damaging agents and to express the embryonal transcription factor OCT4. We recently characterized EC cells that lack OCT4 expression and show cisplatin resistance. In the present, immunohistochemical study we analyzed the composition of NSGCT with the focus on such OCT4-negative EC cells using a NSGCT xenograft model as well as patient-derived NSGCT samples. In the xenograft model, the cisplatin-sensitive cell line H12.1 gives rise to xenografts where EC structures are mainly composed of OCT4-positive cells, whereas xenografts from the resistant cell line 1411HP exclusively comprise OCT4-negative EC areas. We found that post-chemotherapy residual metastatic tumors of patients can be comprised of exclusively OCT4-negative EC, whereas the matched testicular primary tumor harbors OCT4-positive EC. Thorough histological analyses revealed a few examples of such OCT4-negative EC cells also in the testicular primary tumor as well as in xenografts from the cisplatin-sensitive NSGCT-cell line. For these cells we propose an identity as early extraembryonal progenitor cells directly derived from OCT4-expressing EC cells. This challenges the use of the term EC cell. The data also support our hypothesis that malignant growth of resistant NSGCT may be driven by this cell type.

Background: This phase 2 trial was designed to evaluate the efficacy and toxicity of a combination of docetaxel, gemcitabine, and oxaliplatin for platinum- and paclitaxel-pretreated epithelial ovarian cancer., Patients and Methods: Heavily pretreated patients (N = 30; median age, 61 years) received docetaxel, 55 mg/m2; gemcitabine, 500 mg/m2 (day 1); and oxaliplatin, 70 mg/m2 (day 2) biweekly. Twelve patients had platinum-sensitive disease, and 18 patients had platinum-resistant disease., Results: Median follow-up was 18.6 months. No differences in patient characteristics were observed between patients with carboplatinum-sensitive and carboplatinum-resistant disease. In patients with carboplatin-sensitive disease, an overall response (OR) of 83.3%, a progression-free survival of 10.6 months, and an overall survival of 18.9 months were observed. In patients with carboplatinum-resistant disease, an OR was seen in 38.9% with a progression-free survival of 5.3 months and an overall survival of 16.3 months. Patients with platinum-refractory disease (progression under previous carboplatinum therapy, n = 13) had an OR of 23%, whereas patients with objective response but relapse less than 6 months after carboplatinum therapy had an OR of 80.0%. Grade 3 and 4 toxicities were only observed for anemia (6.7%), neutropenia (20.0%), thrombopenia, peripheral neuropathy, and diarrhea (16.7%). No neutropenic fever or treatment-related death occurred., Conclusions: In comparison with current standard protocols, a combination of docetaxel, gemcitabine, and oxaliplatin showed considerably higher efficacy without remarkable increased toxicity; particularly for patients with early relapse after a platinum-containing therapy.

Differentiation and malignant transformation of stem cells are regulated by epigenetic mechanisms. We analyzed promoter methylation and expression of the stem cell determining genes Brachyury, DPPA5, FGF4, FOXD3, LIN28, NESTIN and ZFP42 depending on the differentiation state in human mesenchymal stem cells (MSC), human embryonal carcinoma cells (ECC) and somatic tumor cells. Differentiation of MSC into osteoblasts and adipocytes was accompanied with a loss of expression of the Brachyury gene and downregulation of LIN28. Inactivation of Brachyury was associated with progressive methylation of its CpG island promoter. In ECC promoter methylation of stem cell markers was more frequent in the differentiated subgroup (71%) compared to undifferentiated ECC (29%) and this was associated with downregulation of Brachyury, DPPA5, FGF4, FOXD3, LIN28 and ZFP42. DPPA5 was methylated and NESTIN was unmethylated in most tumor cells. In somatic tumor cells, methylation of stem cell markers (Brachyury, DPPA5, FGF4, FOXD3, LIN28 and ZFP42) was frequently observed (85%). Treatment of cell lines with an inhibitor of DNA methyltransferase reactivated the expression of DPPA5, FGF4, FOXD3, LIN28 and ZFP42, indicating that aberrant promoter methylation is a crucial event that results in their silencing. Our results suggest that epigenetic inactivation of stem cell associated genes is mediated by promoter methylation and that this may represent a fundamental mechanism during normal differentiation processes.

OCT4 is considered a main regulator of embryonic stem cell pluripotency and self renewal capacity. It was shown that relevant OCT4 expression only occurs in cells of embryonic pluripotent nature. However, several recent publications claimed to have demonstrated OCT4 expression in human somatic tumor cells, human adult stem or progenitor cells and differentiated cells.We analysed 42 human tumor cell lines from 13 entities and human bone marrow-derived mesenchymal stem cells (MSC). To validate OCT4 expression we used germ cell tumor (GCT) cell lines, derived xenografts and GCT samples. Analysis by RT-PCR, western blotting, immunocytochemistry and immunohistochemistry was performed. With exception of typical embryonal carcinoma cells, we did not observe reliable OCT4 expression in somatic tumor cell lines and MSC. We suggest that a high level of expression of the OCT4 protein together with its nuclear localization still remains a reliable and definitive feature of cells with embryonic pluripotent nature.

Introduction: The aim of the present study was to develop a test setup with continuous angle alteration to imitate elbow joint motion for the mechanical evaluation of tension band wiring and a newly designed intramedullary nail., Materials and Methods: The servo-pneumatical test stand worked with a rotational angle-adjusted and a linear force-adjusted engine. The fracture model was dynamically tested under cyclic loading imitating elbow joint motion. In total, 14 fresh cadaver upper extremities underwent olecranon fracture by means of transverse osteotomy and were assigned to two groups: tension band wiring and intramedullary nailing. There was a continuous angle alteration between 0 and 1000 of flexion, with continuous changing pull force between 25 N and 150 N. Two steel pins were placed in the proximal, two in the distal olecranon fragment for video analysis of the motion between the two pairs of pins. Displacement in the fracture gap was determined after 4 and 300 cycles., Results: After 300 cycles, the displacement in the fracture fixation model was significantly higher in the tension band wiring group than in the intramedullary nailing group., Discussion: Other studies evaluating biomechanical properties of olecranon osteosyntheses with joint involvement did not change the force direction dynamically. We introduced a test setup with continuous angle alteration to imitate joint motion. This is an important step for accurate biomechanical evaluation of the treatment of different fixation methods in olecranon fractures. The tested nailing system showed significant advantages in loosening under cyclic loading compared to tension band wiring.

Introduction: The aim of this prospective study was to evaluate load-transfer mechanisms and stress patterns of periacetabular cortical and cancellous bone after cemented total hip arthroplasty (THA) in vivo using computed tomography (CT) assisted osteodensitometry. In addition we analyzed the efficacy of CT in detecting radiolucent lines around the acetabular component compared to plain radiography., Materials and Methods: Twenty-two cemented acetabular cups were investigated using conventional sequential axial CT scans (Ø 8 days and 26 months post-OP) and plain radiography (Ø 5 days and 40 months post-OP). CT assisted osteodensitometry was used to determine cancellous and cortical bone bone density (BD). Radiolucent lines were evaluated using both CT and plain radiography., Results: Significant BD loss at the time of follow-up was only detectable ventral to the cup (cortical bone: -16%, P = 0.001; cancellous bone: -31%, P = 0.001). The BD changes dorsal and cranial to the cup were not significant. Postoperatively no radiolucent lines were observed in the cement-bone interface by CT, while on plain radiography acetabular lucent lines were seen in 12 out of 22 cases., Conclusion: CT-osteodensitometry has the technical ability to discriminate between cortical and cancellous bone structures with respect to strain-adapted remodeling: sufficient cancellous and cortical bone stock remained dorsal and cranial to the cup indicative of a balanced load transfer to these regions. CT-osteodensitometry has the potential to become an effective instrument for quality control in THA and the method of choice for in vivo determination of periprosthetic BD. In contrast, plain radiography is more suitable for the early detection of radiolucent lines compared to axial CT scans.

Compartment syndrome of the upper extremity is rare, but happens frequently. It most often affects the forearm, compartment syndromes of the upper arm and hand are seen much more seldom. Early diagnosis and efficient fasciotomy is of highest importance to achieve good outcome and prevent development of Volkmann's ischemic contracture.

Purpose: Open reduction and internal fixation of radial neck fractures can lead to secondary loss of reduction and nonunion due to insufficient stability. Nevertheless, there are only a few biomechanical studies about the stability achieved by different osteosynthesis constructs., Methods: Forty-eight formalin-fixed, human proximal radii were divided into 6 groups according to their bone density (measured by dual-energy x-ray absorptiometry). A 2.7-mm gap osteotomy was performed to simulate an unstable radial neck fracture, which was fixed with 3 nonlocking implants: a 2.4-mm T plate, a 2.4-mm blade plate, and 2.0-mm crossed screws, and 3 locking plates: a 2.0-mm LCP T plate, a 2.0-mm 6x2 grid plate, and a 2.0-mm radial head plate. Implants were tested under axial (N/mm) and torsional (Ncm/ degrees ) loads with a servohydraulic materials testing machine., Results: The radial head plate was significantly stiffer than all other implants under axial as well as under torsional loads, with values of 36 N/mm and 13 Ncm/ degrees . The second-stiffest implant was the blade plate, with values of 20 N/mm and 6 Ncm/ degrees . The weakest implants were the 2.0-mm LCP, with values of 6 N/mm and 2 Ncm/ degrees , and the 2.0-mm crossed screws, with values of 18 N/mm and 2 Ncm/ degrees . The 2.4-mm T plate, with values of 14 N/mm and 4 Ncm/ degrees , and the 2.0-mm grid plate, with values of 8 N/mm and 4 Ncm/ degrees came to lie in the midfield., Conclusions: The 2.0-mm angle-stable plates-depending on their design-allow fixation with comparable or even higher stability than the bulky 2.4-mm nonlocking implants and 2.0-mm crossed screws.

Objective: Toxicity related to the infusion of dimethylsulfoxide (DMSO)-cryopreserved peripheral blood stem cells (DMSO-PBSC) mainly comprises cardiovascular events. Fatal neurotoxicity has been reported in a few cases. DMSO represents the putative causative agent of such rare toxicities and elaborate strategies to replace DMSO would benefit from the identification of predisposing factors for DMSO-related toxicities., Methods: Here, we report on DMSO-related neurotoxicity in a series of patients (n = 51) receiving DMSO-PBSC. The analyzed patient-series included eight patients with pre-existing cerebral disease, partially with a history of epileptic seizures., Results: Neurotoxicity was observed in only one patient who suffered from a generalized tonic seizure upon infusion of DMSO-PBSC and for which the clinical course is reported herein. No neurotoxicity was observed in the group of patients with pre-existing neurological disease. Furthermore, no neurotoxicity was observed in patients who received particularly large volumes of DMSO. In all patients, mild non-neurological side effects occurred but besides the reported seizure no other severe adverse events were observed upon PBSC-infusion., Conclusions: To our knowledge, this is the first report addressing the identification of predisposing factors for DMSO-related neurotoxicty. We conclude that infusion of DMSO-PBSC can be performed safely in patients with pre-existing cerebral disease despite the rare occurrence of severe neurotoxicity. Retrospective multicenter studies are warranted to identify patients who would benefit from elaborate methods of DMSO-replacement.

Aims: At present, clinical success of hepatocyte transplantation as an alternative to whole liver transplantation is hampered by the limited availability of suitable donor organs for the isolation of transplantable hepatocytes. Hence, novel cell sources are required to deliver hepatocytes of adequate quality for clinical use. Mesenchymal stem cells (MSCs) from human bone marrow may have the potential to differentiate into hepatocytes in vitro and in vivo., Methods: Isolated MSCs were selected by density gradient centrifugation and plastic adherence, differentiated in the presence of human hepatocyte growth medium and transplanted in immunodeficient Pfp/Rag2 mice., Results: Here, we demonstrate that human MSCs gain in vitro the characteristic morphology and function of hepatocytes in response to specified growth factors. Specifically, preconditioned MSCs store glycogen, synthesise urea and feature the active hepatocyte-specific gene promoter of phosphoenolpyruvate carboxykinase (PCK1). After transplantation into livers of immunodeficient mice, preconditioned MSCs engraft predominantly in the periportal portion of the liver lobule. In situ, the cells continue to store glycogen and express PCK1, connexin32, albumin and the human hepatocyte-specific antigen HepPar1, indicating that the transplanted cells retain prominent qualities of hepatocytes after their regional integration., Conclusion: MSCs derived from human bone marrow may serve as a novel source for the propagation of hepatocyte-like cells suitable for cell therapy in liver diseases.

For various potential clinical applications, the use of autologous human MSCs (hMSCs) would be favorable. In vitro observations suggested that hMSCs are resistant for chemotherapeutic substances; however, no data exist on the characteristics of hMSCs from bone marrow (BM) of chemotherapeutically treated patients. Here, we analyzed the character of hMSCs derived from chemotherapy-exposed BM and the in vitro response of hMSCs to chemotherapeutic substances. Colony-forming units-fibroblast (CFU-Fs) were isolated from BM aspirates of patients after high-dose or standard chemotherapy and of donors with unaffected BM. CFU-Fs from chemotherapy-exposed and unaffected BM contained hMSCs with similar phenotype, proliferation capacity, and differentiation potential. No obvious influence of age, sex, or time since chemotherapy exposure on the presence and characteristics of hMSCs was observed. In vitro, hMSCs showed a significant resistance for cisplatin, vincristine, and etoposide compared with sensitive tumor cell lines, particularly at apoptosis-inducing doses. The phenotype and differentiation potential of hMSCs was not altered by genotoxic treatment under clinically relevant conditions in vitro. hMSCs showed an elevated threshold for cisplatin-induced apoptosis, which was characterized by a lack of caspase-9 activity in apoptotic cells. In vitro exposure of hMSCs to cisplatin, vincristine, and etoposide resulted in an increased p53 expression, independent of apoptosis induction. We conclude that hMSCs can be isolated from chemotherapy-exposed BM in sufficient number and quality for potential clinical applications in chemotherapeutically treated patients. Our data suggest that an elevated apoptotic threshold contributes not only to the persistence of hMSCs in the BM after chemotherapy but also to their lifelong presence in the adult BM.

Study Design: Computed tomography aided evaluation of spinal decompression by ultrasound-guided spinal fracture repositioning, ligamentotaxis, and remodeling after thoracolumbar burst fractures., Objectives: To determine the necessity of spinal canal widening by ultrasound-guided fracture repositioning for fractures with and without neurologic deficit., Summary of Background Data: Ultrasound-guided spinal fracture repositioning is an alternative new approach. Reports have varied concerning ligamentotaxis and remodeling., Methods: Computed tomography aided planimetry of the spinal canal (64 consecutive burst fractures) and neurologic evaluation by Frankel grades., Results: Ultrasound-guided spinal fracture repositioning (n = 37) reduced the stenosis of the spinal canal area from 45% before surgery to 20% after surgery of the estimated original area. Fifteen patients had a primary neurologic deficit, which improved markedly in 11 cases after treatment. Patients with neurologic symptoms had a greater preoperative spinal stenosis than those without. No correlation was seen between the degree of pretreatment spinal stenosis, fracture type, and severity of the neurologic deficit. Ligamentotaxis (n = 27) reduced the stenosis from 30% before surgery to 18% after surgery and remodeling (n = 11) from 25% after surgery to 13% after metal removal., Conclusion: Ultrasound-guided fracture repositioning is an efficient method for spinal canal decompression of burst fractures with neurologic symptoms. The marked degree of widening of the spinal canal due to the effects of ligamentotaxis and remodeling may render the reposition of retropulsed fragments unnecessary in cases of fractures without a neurologic deficit.

Thirty-six consecutive patients with burst fractures of the thoracolumbar spine and with a fractured posterior vertebral surface dislocated into the spinal canal without neurological symptoms were treated with the AO internal fixator. Computed tomography-aided planimetry of the spinal canal was undertaken preoperatively and within 1 week postoperatively to elucidate the effect of kyphosis correction and distraction on spinal canal widening (ligamentotaxis). The stenosis of the spinal canal area (SCA) was reduced from 29% preoperatively to 19% postoperatively (+10%) of the estimated original area, and the stenosis of the mid-sagittal diameter (MSD) reduced from 31 to 23% (+8%). The widening of the SCA was greater at the level of L1/L2 (+13%) than at L3/L4 (+6%). High preoperative canal compromise was associated with greater absolute spinal canal widening. Large trapezoid-shaped fragments resisted reduction by ligamentotaxis. Even though the effect of ligamentotaxis after operative treatment with the internal fixator was proven, a certain stenosis of the spinal canal remains in most cases. Especially for patients with fracture-related neurological symptoms, ligamentotaxis alone does not seem sufficient for the requested spinal decompression. Even an exact analysis of preoperative CT scans under consideration of the fracture level will not always allow an exact prognosis of the expected effect of ligamentotaxis.

Although the majority of testicular germ cell tumors (TGCTs) are curable by cisplatin-based chemotherapy, in a few cases, the occurrence of cisplatin resistance results in a poor outcome. The biological basis of this differential cisplatin sensitivity in TGCTs remains largely unexplained. Embryonal carcinoma (EC) cells represent the presumptive tumor stem cells in nonseminomatous TGCTs and are known to express the embryonal transcription factor Oct-3/4 and to be hypersensitive to cisplatin. In the present study, we analyzed TGCT cell lines and nude mouse xenografts showing differential cisplatin sensitivity. Here we demonstrate that a lack of expression of Oct-3/4 in TGCT cells is associated with a higher apoptotic threshold and cisplatin resistance which is accompanied by an impaired caspase-9 activation, reduced caspase-3 activity and altered p53 accumulation. We were able to induce loss of Oct-3/4 in a cisplatin-sensitive EC cell line resulting in a secondary cisplatin-resistant cell type with retained EC cell characteristics and changes in apoptotic signaling identical to those in primary resistant cells. Furthermore, we show that EC cells are retained in their undifferentiated state by Oct-3/4 and that a complete and ultimate loss of Oct-3/4 followed by an early differentiation step is necessary to establish the cisplatin-resistant state. Our data suggest that loss of Oct-3/4 expression leads to induction of a higher apoptotic threshold and to cisplatin resistance in EC cells of nonseminomatous TGCTs. We hypothesize that in refractory TGCTs the original tumor stem cell population of Oct-3/4-positive, cisplatin-sensitive EC cells could be replaced by an Oct-3/4-negative, resistant population in a selection process. In contrast, the presence of the Oct-3/4-positive, highly sensitive EC cells as the tumor stem cell component in most TGCTs could explain the general high chemosensitivity and curability of these tumors.

The etiology of chronic lymphocytic leukemia (CLL) appears to be influenced by genetic factors which may contribute to its differential, gender- and age-specific incidence. The presented study is the first, which investigated the frequencies of DNA-typed alleles of all relevant human leukocyte antigens (HLA) loci in CLL patients with regard to gender and age at disease onset. The most remarkable result was the higher frequency of homozygosity for MHC class II loci in female patients. Particularly, an increased frequency of overall homozygosity for the DRB3/4/5 loci was observed in female patients compared to gender matched controls (RR = 2.8) and male patients. The previously demonstrated association of DQB1 homozygosity with CLL in general was found to be specific for female patients (RR = 4.4). Considering the lack of an a priori hypothesis which made it virtually impossible to obtain statistical significance it was not unexpected that none of the observed differences remained significant after correction for multiple comparisons. However, these results suggest a recessive, gender-specific susceptibility factor for CLL within or in vicinity of the human MHC class II region and should serve as an a priori hypothesis for future studies focusing on these gene loci. Furthermore, an increased frequency for HLA-Cw*06 was seen in patients with an early onset age (RR = 2.7) but lost significance after correction for multiple comparisons. The previously reported association of HLA-DRB4*0103 with CLL in general was observed in all groups irrespective of gender and age. Conclusively, our study supports the concept, that CLL represents a disease with a complex etiology and genetic susceptibility which appears to be influenced by the human MHC.

During septic shock, circulating levels of anti-inflammatory mediators are increased relative to those of pro-inflammatory. The reduced capacity of septic shock blood leukocytes in expressing pro-inflammatory genes in response to bacterial lipopolysaccharide endotoxin (LPS) may contribute to reductions in these mediators, but the reasons for persistent increases in circulating anti-inflammatory mediators are unknown. We determined whether septic shock leukocytes that have adapted to LPS induction of the IL-1beta gene could continue to express sIL-1RA in response to LPS. Septic shock whole-blood leukocytes and neutrophils (PMNs) selectively maintained production of sIL-1RA after treatment with LPS while limiting that of IL-1beta. Repressed transcription of IL-1beta and rapid decay of IL-1beta mRNA in septic shock neutrophils correlated with reductions in levels of IL-1beta after stimulation with LPS. Transcription of sIL-1RA mRNA was also suppressed, but the ability of LPS to stimulate events that lead to efficient translation of a stable sIL-1RA mRNA appeared responsible for maintaining sIL-1RA production. We conclude that LPS adaptation of septic shock leukocytes selectively influences signaling pathways that regulate transcription, mRNA processing, and translation, leading to changes in the balance of production of pro- and anti-inflammatory mediators.