Evaluation of Virulence of Salmonella Infantis and Salmonella Enteritidis with In Vitro and In Vivo Models.

Salmonella Infantis and Enteritidis serovars have been reported as important causes of salmonellosis in humans worldwide. However, the virulence of these two serovars has yet to be compared. To evaluate the virulence of Salmonella Infantis (n = 23) and Salmonella Enteritidis (n = 7), we used two models: the Caco2 cells model (in vitro) and the Galleria mellonella model (in vivo). Additionally, the virulence genes of all tested strains were contrasted with phenotypic outcomes. Results showed that adhesion means were 18.2% for Salmonella Enteritidis and 38.2% for Salmonella Infantis strains. Invasion means were 77.1% for Salmonella Enteritidis and 56.2% for Salmonella Infantis strains. Significant differences were found between serovars in adherence and invasion assays. Mortality rates (58% for Salmonella Enteritidis and 62.6% for Salmonella Infantis) were not significantly different between serotypes. The distribution of virulence genes showed that genes fae (fimbrial adherence determinants) and shdA (nonfimbrial adherence determinants) were only found in Salmonella Infantis strains. On the other hand, the rck gene (invasion) and Plasmid-encoded fimbriae genes (pef A, B, C, D) were present in Salmonella Enteritidis exclusively. In conclusion, this study shows that Salmonella Enteritidis has a higher virulence potential under experimental conditions than Salmonella Infantis. However, more studies are needed to determine the risk that Salmonella Infantis could represent compared with Salmonella Enteritidis. Moreover, other in vivo models should be considered to assess the virulence of these serovars.

The impact of type 2 diabetes on bone metabolism and growth after spinal fusion.

BACKGROUND CONTEXT: Some clinical reports suggest diabetes may have a negative effect on spinal fusion outcomes, although no conclusive experimental research has been conducted to investigate the causality, impact, and inherent risks of this growing patient population. PURPOSE: To analyze the hypothesis that type 2 diabetes (T2DM) inhibits the formation of a solid bony union after spinal fusion surgery by altering the local microenvironment at the fusion site through a reduction in growth factors critical for bone formation. STUDY DESIGN/SETTING: In vivo rodent model of type 2 diabetes. METHODS: Twenty control (Sprague Dawley, SD) and 30 diabetic (Zucker Diabetic Sprague Dawley, ZDSD) rats underwent posterolateral and laminar fusion surgery using a tailbone autograft implanted onto the L4/L5 transverse processes. A subset of animals was sacrificed 1-week postsurgery for growth factor analysis. Remaining rats were sacrificed 3-month postsurgery for fusion evaluation via manual palpation, micro-CT, and histology. RESULTS: There was no significant difference in the manual palpation fusion rate between ZDSD rats and SD control rats. Growth factor assay of fusion site explants at early sacrifice demonstrated PDGF was upregulated in the ZDSD rats. TGFB, IGF, and VEGF were not statistically different between groups. Bone mineral density as determined by micro-CT was significantly lower in ZDSD rats compared to SD controls and was a significant function of HbA1c. CONCLUSIONS: Data generated in this in vivo rat model of T2DM demonstrate that the metabolic dysregulation associated with the diabetic condition negatively impacts the quality and density of the formed fusion mass. Increased measures of diabetic status, as determined by blood glucose and HbA1c, were correlated with decreased quality of formed fusion, highlighting the importance of diabetic status monitoring and regulation to bone health, particularly during bone healing. CLINICAL RELEVANCE: T2DM rats demonstrated increased rates of infection, metabolic dysregulation, and a reduction in spinal fusion consolidation. Clinicians should consider these negative effects during preoperative care and treatment of this growing patient population.

Biomechanical Analysis of a Novel Buried Fixation Technique Using Headless Compression Screws for the Treatment of Patella Fractures.

The traditional technique for patella fracture fixation utilizes prominent hardware. Prominent hardware use, however, results in a high rate of reoperation for symptomatic implant removal. This biomechanical study evaluates the effectiveness of a novel patella fixation technique that minimizes implant prominence. Patellar transverse osteotomies were created in 13 pairs of cadaveric knees. Paired knees were assigned to either standard fixation (SF) using cannulated partially threaded screws and stainless steel wire tension band, or buried fixation (BF) using headless compression screws with a No. 2 FiberWire tension band and a No. 5 FiberWire cerclage suture. Quadriceps tendons were cyclically loaded to full extension followed by load to failure. The gap across the fracture site, stiffness, and load to failure were measured. The differences in stiffness and load to failure between the 2 groups were not statistically significant. During cyclic loading, significantly greater gapping was observed across the fracture site in the BF group compared with SF group (P < .05). Both constructs failed under loads that exceeded typical loads experienced during the postoperative rehabilitation period. Nevertheless, the BF technique demonstrated larger gap formation and a reduced load to failure than the SF technique. Further clinical studies are therefore underway to determine whether the use of constructs with decreased stability but increased patient comfort could improve clinical outcomes and reduce reoperation rates.

Effect of modulating dietary vitamin D on the general bone health of rats during posterolateral spinal fusion.

Vitamin D plays a significant role in musculoskeletal health by regulating calcium, phosphate, and promoting new bone mineralization. The purpose of this study was to understand the effect of dietary vitamin D on general bone health during peri-operative bone healing via an in vivo dosing study of vitamin D in a rat posterolateral fusion model using autograft. Vitamin D Deficient (DD), vitamin D Insufficient (ID), Control vitamin D (CD), and Hyper-vitamin D (HD) groups were studied. Increasing dietary vitamin D improved quantitative measures of femoral geometry, including femoral strength, stiffness, and density. Femoral biomechanics, cortical thickness, moment of inertia, cross-sectional area, and measures from bone ashing were all greater in the HD group versus the CD. This suggests that additional dietary vitamin D above normal levels during spinal fusion may lead to improvement in bone health. Serum vitamin D levels were also observed to decrease during fusion healing. These results demonstrate that dietary vitamin D improves general bone health in the femur of a rat model during posterolateral spinal fusion. This suggests a role for further clinical evaluation of vitamin D dietary intake during the peri-operative period, with the possibility of avoiding adverse consequences to general bone health. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1435-1443, 2018.

The effect of a dynamic PCL brace on patellofemoral compartment pressures in PCL-and PCL/PLC-deficient knees.

BACKGROUND: The natural history of posterior cruciate ligament (PCL) deficiency includes the development of arthrosis in the patellofemoral joint (PFJ). The purpose of this biomechanical study was to evaluate the hypothesis that dynamic bracing reduces PFJ pressures in PCL- and combined PCL/posterolateral corner (PLC)-deficient knees. STUDY DESIGN: Controlled Laboratory Study. METHODS: Eight fresh frozen cadaveric knees with intact cruciate and collateral ligaments were included. PFJ pressures and force were measured using a pressure mapping system via a lateral arthrotomy at knee flexion angles of 30°, 60°, 90°, and 120° in intact, PCL-deficient, and PCL/PLC-deficient knees under a combined quadriceps/hamstrings load of 400 N/200 N. Testing was then repeated in PCL- and PCL/PLC-deficient knees after application of a dynamic PCL brace. RESULTS: Application of a dynamic PCL brace led to a reduction in peak PFJ pressures in PCL-deficient knees. In addition, the brace led to a significant reduction in peak pressures in PCL/PLC-deficient knees at 60°, 90°, and 120° of flexion. Application of the dynamic brace also led to a reduction in total PFJ force across all flexion angles for both PCL- and PCL/PLC-deficient knees. CONCLUSION: Dynamic bracing reduces PFJ pressures in PCL- and combined PCL/PLC-deficient knees, particularly at high degrees of knee flexion.

Biomechanical analysis of lateral interbody fusion strategies for adjacent segment degeneration in the lumbar spine.

BACKGROUND CONTEXT: Surgical treatment of symptomatic adjacent segment disease (ASD) typically involves extension of previous instrumentation to include the newly affected level(s). Disruption of the incision site can present challenges and increases the risk of complication. Lateral-based interbody fusion techniques may provide a viable surgical alternative that avoids these risks. This study is the first to analyze the biomechanical effect of adding a lateral-based construct to an existing fusion. PURPOSE: The study aimed to determine whether a minimally invasive lateral interbody device, with and without supplemental instrumentation, can effectively stabilize the rostral segment adjacent to a two-level fusion when compared with a traditional posterior revision approach. STUDY DESIGN/SETTING: This is a cadaveric biomechanical study of lateral-based interbody strategies as add-on techniques to an existing fusion for the treatment of ASD. METHODS: Twelve lumbosacral specimens were non-destructively loaded in flexion, extension, lateral bending, and torsion. Sequentially, the tested conditions were intact, two-level transforaminal lumbar interbody fusion (TLIF) (L3-L5), followed by lateral lumbar interbody fusion procedures at L2-L3 including interbody alone, a supplemental lateral plate, a supplemental spinous process plate, and then either cortical screw or pedicle screw fixation. A three-level TLIF was the final instrumented condition. In all conditions, three-dimensional kinematics were tracked and range of motion (ROM) was calculated for comparisons. Institutional funds (<$50,000) in support of this work were provided by Medtronic Spine. RESULTS: The addition of a lateral interbody device superadjacent to a two-level fusion significantly reduced motion in flexion, extension, and lateral bending (p<.05). Supplementing with a lateral plate further reduced ROM during lateral bending and torsion, whereas a spinous process plate further reduced ROM during flexion and extension. The addition of posterior cortical screws provided the most stable lateral lumbar interbody fusion construct, demonstrating ROM comparable with a traditional three-level TLIF. CONCLUSIONS: The data presented suggest that a lateral-based interbody fusion supplemented with additional minimally invasive instrumentation may provide comparable stability with a traditional posterior revision approach without removal of the existing two-level rod in an ASD revision scenario.

Estrogen preferentially promotes the differentiation of CD11c+ CD11b(intermediate) dendritic cells from bone marrow precursors.

Sex biases in autoimmunity and infection suggest that steroid sex hormones directly modulate immune cells. We show in this study that 17-beta-estradiol (E2) promotes the differentiation of functional dendritic cells (DC) from murine bone marrow precursor cells. Remarkably, ex vivo DC differentiation was inhibited in steroid hormone-deficient medium, and was restored by addition of physiological amounts of E2, but not dihydrotestosterone. DC differentiation was inhibited by the estrogen receptor (ER) antagonists ICI 182,780 and tamoxifen, and from ERalpha(-/-) bone marrow cells, indicating that E2 acted via ERs. E2 addition was most effective in promoting DC differentiation immediately ex vivo, but did not increase DC proliferation. E2 treatment specifically promoted differentiation of a CD11c(+) CD11b(int) DC population that displayed high levels of cell surface MHC class II and CD86, suggesting that E2 could augment numbers of potent APC. DC that differentiated in E2-supplemented medium were fully functional in their capability to mediate presentation of self and foreign Ags and stimulate the proliferation of naive CD4(+) T cells. The requirement for estrogen during DC differentiation suggests a mechanism by which E2 levels in peripheral tissues might modulate both the number and functional capabilities of DC in vivo, thereby influencing immune responses.