Calmodulin Regulated Spectrin Associated Protein 1 mRNA is Directly Regulated by miR-126 in Primary Human Osteoblasts.

Vascularization is essential for bone development, fracture healing, and bone tissue engineering. We have previously described that coculture of primary human osteoblasts (hOBs) and human umbilical vein endothelial cells (HUVECs) improves differentiation of both cell types. Investigating the role of microRNAs (miRNAs) in this system, we found that miR-126 is highly upregulated in hOBs following coculturing with HUVECs. In this study we performed miR-126 gain-of-function and loss-of-function experiments in hOBs followed by microarray analysis in order to identify targets of miR-126. The transcript cluster IDs were sieved by applying cut-off criteria and by selecting transcripts which were upregulated following miR-126 downregulation and vice versa. The calmodulin regulated spectrin associated protein 1 (CAMSAP1) mRNA was confirmed to be differentially regulated by miR-126. Using the luciferase reporter assay it was demonstrated that CAMSAP1 is directly targeted by miR-126. In this study, we show that miR-126 and CAMSAP1 directly interact in hOBs. This finding has potential implications for tissue engineering applications. J. Cell. Biochem. 118: 1756-1763, 2017. © 2016 Wiley Periodicals, Inc.

The lipoprotein receptor LRP1 modulates sphingosine-1-phosphate signaling and is essential for vascular development.

Low density lipoprotein receptor-related protein 1 (LRP1) is indispensable for embryonic development. Comparing different genetically engineered mouse models, we found that expression of Lrp1 is essential in the embryo proper. Loss of LRP1 leads to lethal vascular defects with lack of proper investment with mural cells of both large and small vessels. We further demonstrate that LRP1 modulates Gi-dependent sphingosine-1-phosphate (S1P) signaling and integrates S1P and PDGF-BB signaling pathways, which are both crucial for mural cell recruitment, via its intracellular domain. Loss of LRP1 leads to a lack of S1P-dependent inhibition of RAC1 and loss of constraint of PDGF-BB-induced cell migration. Our studies thus identify LRP1 as a novel player in angiogenesis and in the recruitment and maintenance of mural cells. Moreover, they reveal an unexpected link between lipoprotein receptor and sphingolipid signaling that, in addition to angiogenesis during embryonic development, is of potential importance for other targets of these pathways, such as tumor angiogenesis and inflammatory processes.

miR-126 modulates angiogenic growth parameters of peripheral blood endothelial progenitor cells.

Vascularization plays an important role in tissue engineering applications. It is known that implantation of differentiated endothelial cells or endothelial progenitor cells (EPCs) from cord blood (cbEPCs) gives rise to the formation of a complex functional neovasculature, whereas EPCs isolated from peripheral blood (pbEPCs) have a limited capability to form blood vessels upon implantation. MicroRNA-126 (miR-126) has been shown to have pro-angiogenic effects in vivo. In this study, we investigated whether modulation of miR-126 expression in pbEPCs may alter their angiogenic properties. Gain of function and loss of function experiments revealed that miR-126 has anti-angiogenic effects in pbEPCs. Overexpression of miR-126 resulted in decreased proliferation, migration, invasion and tube formation, while inhibition of miR-126 induced the opposite effects. However, modulation of miR-126 expression did not influence apoptotic susceptibility of pbEPCs. This study provides evidence that inhibition of miR-126 improves angiogenesis-related growth parameters in pbEPCs and may represent a therapeutic option to ameliorate the angiogenic and vasculogenic properties of pbEPCs.

Endothelial progenitor cells from peripheral blood support bone regeneration by provoking an angiogenic response.

Neovascularization is crucial for fracture healing and plays an important role in long-time graft survival in tissue engineering applications. Endothelial progenitor cells (EPCs) can be isolated from peripheral blood avoiding donor site morbidity, which makes them attractive for autologous cell-based engineering of neovessels. However, contradictory results are published concerning the vasculogenic potential of this cell type. We could previously show that implanted human endothelial vein cells (HUVECs) gave rise to the formation of a complex functional human neovasculature in a heterotopic (subcutaneous) as well as in an orthotopic (calvarial defect) model of severe combined immunodeficiency (SCID) mice, where vessel formation could even be increased by coimplanting mesenchymal stem cells (MSCs) functioning as perivascular cells. In this study, we investigated whether coimplantation of MSCs which have been predifferentiated in vitro into SMCs (SMC-MSCs) may enable pbEPCs to form blood vessels upon implantation and, if this would be the case, whether the resulting enhanced vascularization may support bone regeneration. For this purpose, pbEPCs and SMC-MSCs were mono- or cocultured in collagen matrices and seeded into scaffolds consisting of decalcified processed bovine cancellous bone (PBCB, Tutobone). Neovascularization and osteogenesis were evaluated using a calvarial bone defect-model in SCID mice. Our experiments could show that the missing vasculogenic potential of pbEPCs is not rescued by coimplantation of SMCs derived from MSCs predifferentiated along the vascular smooth muscle lineage. However, implantation of both cell types alone, or in combination induced an angiogenic response, which correlated in a positive manner with bone formation within the implants.

The quality of aesthetic surgery training in plastic surgery residency: a survey among residents in Germany.

BACKGROUND: The importance of providing high-quality exposure and training in aesthetic and reconstructive surgery during residency has come to the forefront of plastic surgery education. Adequate training in aesthetic surgery, however, has traditionally been challenging. The authors were interested in how these challenges were met abroad. METHODS: A 17-item online survey was developed to assess the quality of training in aesthetic surgery in Germany. The survey had 3 distinct sections: demographic information, current state of aesthetic surgery training, and residents' opinions about the perceived quality of aesthetic surgery training. Only responses of senior residents were included in the final analysis. RESULTS: A total of 112 residents responded (30% response rate), of which 88 were senior plastic surgery residents. Ninety percent (n = 79) reported that a resident aesthetic surgery clinic was not part of their training experience. Eighty-eight percent (n = 77) reported that they did not have a dedicated aesthetic surgery rotation during their residency training. According to 69.3% (n = 61), no didactic training in aesthetic surgery was provided. Fifty-six percent (n = 49) of senior plastic surgery residents had performed only a maximum of 10 aesthetic surgery procedures at the time of the survey. Although only 43.2% of senior residents claimed to be interested in a predominantly aesthetic surgery practice, 90.9% (n = 80) felt that they require further training in aesthetic surgery (ie, fellowship). CONCLUSIONS: Deficiencies exist with respect to aesthetic surgery training among senior plastic surgery residents in Germany. Structural improvements in residency training with formal exposure and teaching in aesthetic surgery are warranted. The German Society of Plastic, Reconstructive and Aesthetic Surgeons is actively addressing deficiencies identified with the goal of improving the quality of training.

Novel variant of reversed midgut rotation - retro-arterial proximal jejunum and transverse colon: a case report and review of the literature.

BACKGROUND: Reversed rotation of the midgut is the rarest variation of midgut malrotations, which are congenital disorders that result from aberrant rotation and fixation of the midgut during embryological development. Common complications of these disorders are small bowel obstruction by volvulus or peritoneal bands, usually occurring in early infancy. CASE PRESENTATION: A 23-year-old Caucasian woman presented with recurrent abdominal pain. A contrast-enhanced multidetector computed tomography study revealed a novel variant of reversed rotation of the midgut. Besides the specific finding of a retro-arterial transverse colon, we also found the proximal jejunum to cross posterior to the mesenteric root, a variation that has not been reported in the literature so far. In this case, substantial symptomatic relief was achieved with conservative management. CONCLUSIONS: The hypothesis of a double reversed rotation of the pre-arterial segment of the umbilical loop around the superior mesenteric artery axis provides a possible explanation for this anomaly. There is no evidence-based consensus on the management of patients presenting with non-symptomatic or mildly symptomatic intestinal malrotations. In this case, radiologic and clinical presentations excluded acute small bowel obstruction, and surgical intervention was avoided.

Comparison of Diagnostic Accuracy of Radiation Dose-Equivalent Radiography, Multidetector Computed Tomography and Cone Beam Computed Tomography for Fractures of Adult Cadaveric Wrists.

PURPOSE: To compare the diagnostic accuracy of radiography, to radiography equivalent dose multidetector computed tomography (RED-MDCT) and to radiography equivalent dose cone beam computed tomography (RED-CBCT) for wrist fractures. METHODS: As study subjects we obtained 10 cadaveric human hands from body donors. Distal radius, distal ulna and carpal bones (n = 100) were artificially fractured in random order in a controlled experimental setting. We performed radiation dose equivalent radiography (settings as in standard clinical care), RED-MDCT in a 320 row MDCT with single shot mode and RED-CBCT in a device dedicated to musculoskeletal imaging. Three raters independently evaluated the resulting images for fractures and the level of confidence for each finding. Gold standard was evaluated by consensus reading of a high-dose MDCT. RESULTS: Pooled sensitivity was higher in RED-MDCT with 0.89 and RED-MDCT with 0.81 compared to radiography with 0.54 (P = < .004). No significant differences were detected concerning the modalities' specificities (with values between P = .98). Raters' confidence was higher in RED-MDCT and RED-CBCT compared to radiography (P < .001). CONCLUSION: The diagnostic accuracy of RED-MDCT and RED-CBCT for wrist fractures proved to be similar and in some parts even higher compared to radiography. Readers are more confident in their reporting with the cross sectional modalities. Dose equivalent cross sectional computed tomography of the wrist could replace plain radiography for fracture diagnosis in the long run.

Comparison of Multidetector Computed Tomography and Flat-Panel Computed Tomography Regarding Visualization of Cortical Fractures, Cortical Defects, and Orthopedic Screws: A Phantom Study.

To compare the visualization of cortical fractures, cortical defects, and orthopedic screws in a dedicated extremity flat-panel computed tomography (FPCT) scanner and a multidetector computed tomography (MDCT) scanner.We used feet of European roe deer as phantoms for cortical fractures, cortical defects, and implanted orthopedic screws. FPCT and MDCT scans were performed with equivalent dose settings. Six observers rated the scans according to number of fragments, size of defects, size of defects opposite orthopedic screws, and the length of different screws. The image quality regarding depiction of the cortical bone was assessed. The gold standard (real number of fragments) was evaluated by autopsy.The correlation of reader assessment of fragments, cortical defects, and screws with the gold standard was similar for FPCT and MDCT. Three readers rated the subjective image quality of the MDCT to be higher, whereas the others showed no preferences.Although the image quality was rated higher in the MDCT than in the FPCT by 3 out of 6 observers, both modalities proved to be comparable regarding the visualization of cortical fractures, cortical defects, and orthopedic screws and of use to musculoskeletal radiology regarding fracture detection and postsurgical evaluation in our experimental setting.

[Application possibilities and initial experience with digital volume tomography in hand and wrist imaging]. / Digitale Volumentomografie (DVT) des knöchernen Handskeletts: Erste Erfahrungen und Anwendungsmöglichkeiten.

During the last decade, DVT (digital volume tomography) imaging has become a widely used standard technique in head and neck imaging. Lower radiation exposure compared to conventional computed tomography (MDCT) has been described. Recently, DVT has been developed as an extremity scanner and as such represents a new imaging technique for hand surgery. We here describe the first 24 months experience with this new imaging modality in hand and wrist imaging by presenting representative cases and by describing the technical background. Furthermore, the method's advantages and disadvantages are discussed with reference to the given literature.

Comparing the image quality of a mobile flat-panel computed tomography and a multidetector computed tomography: a phantom study.

OBJECTIVES: The aim of this study was to compare the image quality of a compact mobile flat-panel computed tomography (FPCT) capable of extremity imaging and a multidetector computed tomography (MDCT) in examinations with the same radiation dose. MATERIAL AND METHODS: Imaging with the FPCT was performed with default settings. Monte Carlo simulations were used to calculate equivalent dose settings for the 320-row MDCT. Simulations were based on and validated by dose measurements. Homogeneity, geometric distortion, artifacts, accuracy of Hounsfield values, contrast, and spatial resolution were evaluated in different imaging phantoms. Whitney-Mann U Test and Spearman ρ were used for statistical analysis. RESULTS: Homogeneity reached 2.5% for the FPCT and 0.5% for the MDCT. Hounsfield values were more accurate and contrast to noise ratios were higher for the MDCT than the FPCT (P ≤ 0.001). The MDCT depicted more rod inserts than the FPCT did. No significant geometric distortion was detected in either modality. The FPCT was more prone to artifacts around Krischner wires with a diameter of 2 mm (P = 0.05-0.001), whereas the MDCT showed a higher amount of artifacts around wires with a diameter of 0.8 mm (P ≤ 0.001). Spatial resolution was 1 lp/mm (xy), 1.7 lp/mm (z) for the FPCT and 1 lp/mm (xy), less than 1 lp/mm (z) for the MDCT. CONCLUSIONS: We compared a mobile FPCT and a 320-row MDCT by using the same radiation dose for scans. We found the spatial resolution to be higher in the FPCT. Hounsfield units were more accurate and homogeneity and contrast resolution were better in MDCT. The MDCT was also less prone to artifacts from thick Kirschner wires but showed comparably more artifacts around thin wires.

An integrated therapy concept for reduction of postoperative complications after resection of a panniculus morbidus.

BACKGROUND: Panniculus morbidus is characterized by an edematous, painful hanging abdominal mass, due to laxity and redundancy of the abdominal skin in morbid obesity, particularly after massive weight loss. Panniculectomy, by wedge resection, is a salvage procedure with high satisfaction rates though associated with high complication rates. Here we investigated the effects of perioperative complex decongestive physical therapy (CDP) on outcome and complication rates. METHODS: We retrospectively analyzed the clinical course and outcome of 24 patients receiving panniculectomy between 1998 and 2009 in our department of plastic surgery. Sixteen patients received perioperative CDP, and eight patients did not receive any form of decongestive treatment. We analyzed the incidence of complications, reoperation, blood transfusions, and length of hospital stay based on chart reviews. Complications were categorized as minor or major according to the necessity of readmission or reoperation. CDP was performed for 4-6 weeks preoperatively and 2 weeks postoperatively. RESULTS: The incidence of major complications (p = 0.001), the rate of postoperative blood transfusions (p = 0.028), wound healing disorders (p = 0.021), and the incidence for complications (p = 0.001), whether minor or major, were significantly reduced in the CDP group. In summary, 12 of 16 patients within the CDP group had an uneventful course, whereas all non-CDP patients had at least one complication. CONCLUSIONS: Adequate perioperative CDP treatment in a lymphological clinic may reduce the rate of early postoperative complications after resection of panniculus morbidus.

Human endothelial progenitor cells induce extracellular signal-regulated kinase-dependent differentiation of mesenchymal stem cells into smooth muscle cells upon cocultivation.

Neovascularization represents an important issue in tissue-engineering applications, since survival of implanted cells strongly relies on sufficient oxygen and nutrient supply. We have recently observed that human bone marrow-derived mesenchymal stem cells (MSCs) support neovessel formation originating from coimplanted endothelial cells (ECs) in vivo, suggesting that MSCs may function as perivascular cells by investing and stabilizing nascent EC-derived neovessels. In this study, we investigated EC-induced mural cell differentiation of MSCs in vitro. For this purpose, endothelial progenitor cells (EPCs) from two different origins, namely adult peripheral blood (pbEPCs) and neonatal cord blood (cbEPCs), or human umbilical vein endothelial cells (HUVECs), were cocultured with human MSCs to analyze the effect on MSC differentiation toward a smooth muscle cell (SMC)/pericyte phenotype. EPCs as well as HUVECs increased alpha-smooth muscle actin expression in MSCs upon cocultivation in a time-dependent manner. This effect was strongly dependent on direct cell-to-cell contact and extracellular signal-regulated kinase (ERK) signaling, but was not mediated by heterotypic gap junction communication. Beyond enhanced SMC marker gene expression in MSCs, EPCs also enhanced the functional characteristics of cocultured MSCs by increasing their ability to attach to EC tubes in vitro. In conclusion, our study has shown that EPCs from adult peripheral blood as well as from cord blood commit cocultivated MSCs toward an SMC/pericyte phenotype in a cell-contact- and ERK-dependent manner.