Lumbar reconstruction with hyperlordotic cages: Prediction of neuroforaminal height in comparison to established age and sex dependent reference values.

Interbody cages are routinely used in lumbar reconstruction surgery of deformity cases for restoration of lordosis and sagittal balance of the spine. However, if hyperlordotic implants are inserted into the intervertebral space, special consideration has to be taken concerning the height of the neural foramen during cage implantation. The greater the lordotic angle of the cage is, the higher the posterior size of the cage needs to be in order to avoid neuroforaminal nerve root impingement. In this technical communication, we propose and clinically validate a stepwise mathematic model to predict neuroforaminal height in patients undergoing lumbar reconstruction with hyperlordotic cages. The length of the superior and inferior vertebral end plates including the height of the neural foramen are measured before implantation of the cage in standing sagittal view x-rays. By assumption of an isosceles triangle in combination with the posterior height and the lordotic angle of the cage, the neuroforaminal height after cage implantation can be estimated. By comparison of the predicted neuroforaminal height with age and sex dependent reference values, nerve root impingement can be avoided by selection of the necessary posterior height of the hyperlordotic cage while still gaining sufficient lumbar lordosis.

Development and validation of a practical solution for detecting motion artefacts in the EOS X-ray system.

The EOS™2D/3D system is a low-dose, 3D imaging system that utilizes two perpendicular X-ray beams to create simultaneous frontal and lateral images of the body. This is a useful modality to assess spinal pathologies. However, due to the slow imaging acquisition time up to 25 s, motion artifacts (MA) frequently occur. These artifacts may not be distinguishable from pathological findings, such as scoliosis, and may impair the diagnostic process. The aim of this study was to design a method to detect MA in EOS X-ray. We retrospectively analyzed EOS imaging from 40 patients wearing a radiopaque reference device during imaging. We drew a straight vertical line along the reference device. We measured deviations from it to quantify MA, presenting these findings through descriptive statistics. For a subset of patients with high MA, acquisitions were repeated after giving specific instructions to stand still. For these patients, we compared MA between the two acquisitions. In our study, a substantial proportion of patients exhibited MA ≥ 1 mm, with 80% in frontal projections and 87.9% in lateral projections. In the subjects who received a second acquisition, MA was significantly lower in the second images. Our method allows for a precise detection of MA on EOS images through a simple, yet reliable solution. Our method may improve the reliability of spine measurements, and reduce the risk of wrong diagnosis due to low imaging quality.

Multi-rod fixation in spinal neuroarthropathy: a novel surgical technique.

Spinal cord injury (SCI) leads to compromised biomechanical stability due to impaired neuroprotection. This may trigger deformity and destruction of multiple segments of the spine which is known as spinal neuroarthropathy (SNA) or Charcot arthropathy. Surgical treatment of SNA is highly demanding in terms of reconstruction, realignment, and stabilization. In particular, construct failure due to the combination of high shear forces and reduced bone mineral density in the lumbosacral transition zone is a frequent complication in SNA. Notably, up to 75% of SNA patients need multiple revisions within the first year after surgery in order to achieve successful bony fusion. The purpose of this technical report is to present a novel surgical approach with higher overall construct stability to efficiently treat SNA and avoiding repetitive revisions. The new technique of triple rod stabilisation of the lumbosacral transition zone in combination with the introduction of tricortical laminovertebral (TLV) screws is demonstrated in three patients with complete SCI of the thoracic spinal cord. After surgery all patients reported an improvement of the Spinal Cord Independence Measure III (SCIM III) and none of the reported cases showed construct failure within an at least 9 months follow up period. Although TLV screws violate the integrity of the spinal canal, there were no complications with regard to cerebral spinal fluid fistulas and/or arachnopathies so far. The new concept of triple rod stabilization in combination with TLV screws provides improved construct stability in patients with SNA and thus could help to reduce revision and complications rates and improve patient outcome in this disabling degenerative disease.

Spinal cord tethering and syringomyelia after trauma: impact of age and surgical outcome.

Posttraumatic spinal cord tethering and syringomyelia frequently lead to progressive neurological loss. Although several studies demonstrated favourable outcome following spinal cord detethering with/without shunting, additional research is required as no clear consensus exists over the ideal treatment strategy and knowledge about prognostic demographic determinants is currently limited. In this investigation, we retrospectively investigated 67 patients (56 men, 11 women) who were surgically treated and followed for symptomatic spinal cord tethering and syringomyelia from 2012 to 2022 at our center. Age (B-coefficient 0.396) and severity of trauma to the spinal cord (B-coefficient - 0.462) have been identified as independent predictors for the rate of development of symptomatic spinal cord tethering and syringomyelia (p < 0.001). Following untethering surgery including expansion duraplasty with/without shunting, 65.9% of patients demonstrated an improvement of neurological loss (p < 0.001) whereas 50.0% of patients displayed amelioration of spasticity and/or neuropathic pain (p < 0.001). Conclusively, active screening for symptomatic spinal cord tethering and syringomyelia, particularly in younger patients with severe spinal trauma, is crucial as surgical untethering with/without shunting is able to achieve favourable clinical outcomes. This knowledge may enable clinicians to tailor treatment strategies in spinal cord injury patients suffering from progressive neurological loss towards a more optimal and personalized patient care.

Autofluorescence is a Reliable in vitro Marker of Cellular Senescence in Human Mesenchymal Stromal Cells.

Mesenchymal stromal cells (MSC) are used in cell therapies, however cellular senescence increases heterogeneity of cell populations and leads to uncertainty in therapies' outcomes. The determination of cellular senescence is time consuming and logistically intensive. Here, we propose the use of endogenous autofluorescence as real-time quantification of cellular senescence in human MSC, based on label-free flow cytometry analysis. We correlated cell autofluorescence to senescence using senescence-associated beta-galactosidase assay (SA-ß-Gal) with chromogenic (X-GAL) and fluorescent (C12FDG) substrates, gene expression of senescence markers (such as p16INK4A, p18INK4C, CCND2 and CDCA7) and telomere length. Autofluorescence was further correlated to MSC differentiation assays (adipogenesis, chondrogenesis and osteogenesis), MSC stemness markers (CD90/CD106) and cytokine secretion (IL-6 and MCP-1). Increased cell autofluorescence significantly correlated with increased SA-ß-Gal signal (both X-GAL and C12FDG substrates), cell volume and cell granularity, IL-6/MCP-1 secretion and with increased p16INK4A and CCND2 gene expression. Increased cell autofluorescence was negatively associated with the expression of the CD90/CD106 markers, osteogenic and chondrogenic differentiation potentials and p18INK4C and CDCA7 gene expression. Cell autofluorescence correlated neither with telomere length nor with adipogenic differentiation potential. We conclude that autofluorescence can be used as fast and non-invasive senescence assay for comparing MSC populations under controlled culture conditions.

VEGF vascularization pathway in human intervertebral disc does not change during the disc degeneration process.

OBJECTIVE: During degeneration of the intervertebral disc ingrowth of blood vessels and nerves into the disc are associated with back pain. Vascular endothelial growth factors promote vasculogenesis by binding to the membrane vascular endothelial growth factor receptor 1, while shorter soluble forms of this receptor can inhibit vascularization. We hypothesized that membrane and soluble receptor forms might change between stages of intervertebral disc degeneration. RESULTS: Expression of soluble and membrane forms of vascular endothelial growth factor receptor 1 in human degenerated intervertebral discs and healthy bovine caudal discs was assessed by qRT-PCR and immunoblot. Comparative microarray meta-analysis across disc degeneration grades showed that membrane and soluble forms of this receptor, together with other components of classic vascularization pathways, are constitutively expressed across human disc degeneration stages. Contrary to our hypothesis, we observed that expression of the classic vascularization pathway is stable across degeneration stages and we assume that soluble vascular endothelial growth factor receptor 1 does not contribute to prevent disc degeneration. However, we observed increased expression levels of genes involved in alternative vascularization signalling pathways in severely degenerated discs, suggesting that abnormal vascularization is part of the pathological progression of disc degeneration.

Oxidative status predicts quality in human mesenchymal stem cells.

BACKGROUND: Human bone marrow-derived mesenchymal stem cells (MSC) are adult progenitor cells with great potential for application in cell-based therapies. From a cell-based therapy perspective, there are two limitations to MSC use: (1) these therapies require large numbers of cells, and long-term expansion of MSC in vitro promotes replicative senescence; and (2) patient variability is a challenge for defining MSC quality standards for transplantation. This study aimed to determine whether low or high oxidative status of MSC correlate with changes in cell expansion and differentiation potentials. METHODS: We investigated functional aspects of mitochondria, such as cell metabolic activity indicators and expression of antioxidant enzymes. Furthermore, we tested if senescence-induced changes in oxidative status of MSC could be counteracted by methylene blue (MB), an alternative mitochondrial electron transfer known to enhance cell bioenergetics. RESULTS: MSC isolated from donors of the same age showed distinctive behavior in culture and were grouped as weak (low colony-forming units (CFU) and a short life in vitro) and vigorous MSC (high CFU and a long life in vitro). In comparison to weak MSC, vigorous MSC had oxidative status characterized by lower mitochondrial membrane potential, lower mitochondrial activity, and fewer reactive oxygen species production, as well as reduced mitochondrial biogenesis. Vigorous MSC had a significantly higher expansion potential compared to weak MSC, while no differences were observed during differentiation. MB treatment significantly improved expansion and differentiation potential, however only in vigorous MSC. CONCLUSIONS: Together, these results demonstrate the importance of mitochondrial function in MSC in vitro, and that cells with low oxidative status levels are better candidates for cell-based therapies.

Increased motility of mesenchymal stem cells is correlated with inhibition of stimulated peripheral blood mononuclear cells in vitro.

Immunomodulatory properties of mesenchymal stem cells (MSC) are key components of their successful applications in clinical setting. However, treatments based on MSC immunomodulation need understanding of cell characteristics before cell transplantation. We used live-imaging to test the suitability of the MSC motility as a parameter for quick prediction of the immunomodulatory potential of human MSC in regulating the activity of stimulated peripheral blood mononuclear cells (PBMC) in vitro. Bone marrow MSC, from various donors and in vitro passages, were cultured with or without stimulated PBMC. After seven days, immunomodulation was assessed by measuring PBMC proliferation, IgG production and cytokine secretion in MSC and PBMC monocultures and co-cultures, and results were correlated to MSC motility. In co-culture, we observed that MSC successfully inhibited PBMC activity, reducing PBMC proliferation and IgG production compared to PBMC monoculture. MSC modulated PBMC to reduce the secretion of TNFα and IL-10, increase IL-6, G-CSF and MCP-1, while GM-CSF was not affected. By live-imaging tracking of cell trajectories, we observed that fast moving MSC were inhibiting more efficiently stimulated PBMC compared to slow ones. In co-culture, fast MSC were more effective in inhibiting IgG production (˜30% less IgG), and secreted higher levels of IL-10 (˜10% increase) and GM-CSF (˜20% increase) compared to slower cells. Furthermore, fast MSC in monocultures produced 2.3-fold more IL-6, 1.5-fold MCP-1 and 1.2-fold G-CSF in comparison to slower cells. In conclusion, live-imaging cell tracking allowed us to develop an indicative assay of the immune-regulatory potential of MSC prior to in vivo administration. Key Words: Human mesenchymal stem cells, Immunomodulatory potential, In vitro cell motility, Stem cell transplantation.

Charcot arthropathy of the spine in spinal cord injured individuals with sacral deafferentation and anterior root stimulator implantation.

AIMS: To investigate the occurrence of Charcot spinal arthropathy (CSA) after sacral deafferentation (SDAF) and sacral anterior root stimulation (SARS) of the bladder in patients suffering from neurogenic lower urinary tract dysfunction (NLUTD) as a result of spinal cord injury (SCI). METHODS: Retrospective evaluation of patients who had undergone SDAF/SARS at a single SCI rehabilitation centre. The occurrence rate of stimulation dysfunction was determined, and the medical records and radiological images of the included patients were examined for CSA. The diagnosis of CSA was based on radiological criteria. The occurrence rate of CSA was estimated for all SARS patients and for those with SARS dysfunction, and the odds ratios (OR) for the occurrence of CSA were calculated. RESULTS: In 11/130 SARS patients (8%), CSA was observed a median 8 years (95% CI 5-16 years) after SDAF/SARS or a median 21 years (95% CI 9-41 years) after SCI had occurred. The median follow-up time was 14 years (range 6-25 years). The proportion of patients with CSA was significantly (P = 0.036) greater in patients with SARS dysfunction (7/41) than in patients without SARS dysfunction (4/89). The odds of CSA were four times greater (OR 4.3, 95% CI 1.0-21.5) in patients with SARS dysfunction compared to those without. Furthermore, the odds of CSA were 20 times greater (OR 20.2, 95% CI 8.4-47.0) in patients with SARS compared to those without. CONCLUSIONS: Charcot spinal arthropathy should be considered a potential long-term complication of SDAF/SARS, and spinal instability is a possible reason for SARS dysfunction.

Growth Factors Cross-Linked to Collagen Microcarriers Promote Expansion and Chondrogenic Differentiation of Human Mesenchymal Stem Cells.

Tissue engineering is a field in progressive expansion and requires constant updates in methods and devices. One of the central fields is the development of biocompatible, biodegradable, and injectable scaffolds, such as collagen microcarriers. To enhance cell attachment and produce a cost-effective cell culture solution with local stimulation of cells, basic fibroblast growth factor (bFGF) or transforming growth factor-ß1 (TGF-ß1) was covalently immobilized on microcarriers either by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) or riboflavin/UV (RB/UV) light-mediated cross-linking. Collagen microcarriers cross-linked with bFGF or TGF-ß1 were used for expansion and chondrogenic differentiation of human mesenchymal stem cells (MSCs). Evaluation methods included cell viability test, chondrogenic marker expression (aggrecan and collagen type I and type II), histological detection of proteoglycans, and immunohistochemical analysis. Cross-linking strengthened the collagen structure of the microcarriers and reduced collagenase-mediated degradation. MSCs effectively proliferated on microcarriers cross-linked with bFGF, especially by EDC/NHS cross-linking. Chondrogenic differentiation of MSCs was induced by TGF-ß1 cross-linked on microcarriers, promoting gene expression and protein accumulation of aggrecan and collagen type I and type II, as well as proteoglycans. Cross-linking by RB/UV enhanced chondrogenesis more than any other group. In addition, cross-linking reduced scaffold shrinkage exerted by MSCs during chondrogenesis, a desirable feature for microcarriers if used as tissue defect filler. In conclusion, cross-linking of bFGF or TGF-ß1 to collagen microcarriers supported in vitro proliferation and chondrogenesis, respectively. If translated in vivo and in clinical practice, such approach might lead a step closer to development of a cost-effective and locally acting device for cell-based therapy.

In vitro cell motility as a potential mesenchymal stem cell marker for multipotency.

Mesenchymal stem cells (MSCs) are expected to have a fundamental role in future cell-based therapies because of their high proliferative ability, multilineage potential, and immunomodulatory properties. Autologous transplantations have the "elephant in the room" problem of wide donor variability, reflected by variability in MSC quality and characteristics, leading to uncertain outcomes in the use of these cells. We propose life imaging as a tool to characterize populations of human MSCs. Bone marrow MSCs from various donors and in vitro passages were evaluated for their in vitro motility, and the distances were correlated to the adipogenic, chondrogenic, and osteogenic differentiation potentials and the levels of senescence and cell size. Using life-image measuring of track lengths of 70 cells per population for a period of 24 hours, we observed that slow-moving cells had the higher proportion of senescent cells compared with fast ones. Larger cells moved less than smaller ones, and spindle-shaped cells had an average speed. Both fast cells and slow cells were characterized by a low differentiation potential, and average-moving cells were more effective in undergoing all three lineage differentiations. Furthermore, heterogeneity in single cell motility within a population correlated with the average-moving cells, and fast- and slow-moving cells tended toward homogeneity (i.e., a monotonous moving pattern). In conclusion, in vitro cell motility might be a useful tool to quickly characterize and distinguish the MSC population's differentiation potential before additional use.

Characteristics and surgical management of neuropathic (Charcot) spinal arthropathy after spinal cord injury.

BACKGROUND CONTEXT: Neuropathic (Charcot) spinal arthropathy (CSA) is a rare but progressive and severe degenerative disease that develops in the absence of deep sensation, for example, after spinal cord injury. The diagnosis of CSA is often delayed as a result of the late onset or slow progression of the disease and the nonspecific nature of the reported clinical signs. Considering risk factors of CSA in combination with the common clinical signs may facilitate timely diagnosis and prevent severe presentation of the disease. However, there is a lack of data concerning the early signs and risk factors of CSA. Furthermore, the complications and outcomes after surgical treatment are documented insufficiently. PURPOSE: To investigate the early signs and risk factors of CSA after spinal cord injury, as well as the complications and outcome after surgical treatment. STUDY DESIGN: Retrospective case series from a single center. PATIENT SAMPLE: Twenty-eight patients with 39 Charcot joints of the spine. OUTCOME MEASURES: Clinical signs, radiological signs, risk factors, and complications. METHODS: The case histories and radiological images of patients suffering from CSA were investigated. RESULTS: The first clinical symptoms included spinal deformity, sitting imbalance, and localized back pain. Long-segment stabilization, laminectomy, scoliosis, and excessive loading of the spine were identified as risk factors for the development of the disease. Postoperative complications included implant loosening, wound healing disturbance, and development of additional Charcot joints. All patients were able to return to their previous levels of activities. CONCLUSIONS: Radiological follow-up of the entire thoracic and lumbar spine should be performed in paraplegic patients. Risk factors in combination with typical symptoms should be considered to facilitate early detection. Functional restoration can be achieved with appropriate surgical techniques.

Outcome analysis of retrograde nailing and less invasive stabilization system in distal femoral fractures: A retrospective analysis.

BACKGROUND: TWO MAJOR THERAPEUTIC PRINCIPLES CAN BE EMPLOYED FOR THE TREATMENT OF DISTAL FEMORAL FRACTURES: retrograde intramedullary (IM) nailing (RN) or less invasive stabilization on system (LISS). Both operative stabilizing systems follow the principle of biological osteosynthesis. IM nailing protects the soft-tissue envelope due to its minimally invasive approach and closed reduction techniques better than distal femoral locked plating. The purpose of this study was to evaluate and compare outcome of distal femur fracture stabilization using RN or LISS techniques. MATERIALS AND METHODS: In a retrospective study from 2003 to 2008, we analyzed 115 patients with distal femur fracture who had been treated by retrograde IM nailing (59 patients) or LISS plating (56 patients). In the two cohort groups, mean age was 54 years (17-89 years). Mechanism of injury was high energy impact in 57% (53% RN, 67% LISS) and low-energy injury in 43% (47% RN, 33% LISS), respectively. Fractures were classified according to AO classification: there were 52 type A fractures (RN 31, LISS 21) and 63 type C fractures (RN 28, LISS 35); 32% (RN) and 56% (LISS) were open and 68% (RN) and 44% (LISS) were closed fractures, respectively. Functional and radiological outcome was assessed. RESULTS: Clinical and radiographic evaluation demonstrated osseous healing within 6 months following RN and following LISS plating in over 90% of patients. However, no statistically significant differences were found for the parameters time to osseous healing, rate of nonunion, and postoperative complications. The following complications were treated: hematoma formation (one patient RN and three patients LISS), superficial infection (one patient RN and three patients LISS), deep infection (2 patients LISS). Additional secondary bone grafting for successful healing 3 months after the primary operation was required in four patients in the RN (7% of patients) and six in the LISS group (10% of patients). Accumulative result of functional outcome using the Knee and Osteoarthritis Outcome (KOOS) score demonstrated in type A fractures a score of 263 (RN) and 260 (LISS), and in type C fractures 257 (RN) and 218 (LISS). Differences between groups for type A were statistically insignificant, statistical analysis for type C fractures between the two groups are not possible, since in type C2 and C3 fractures only LISS plating was performed. CONCLUSION: Both retrograde IM nailing and angular stable plating are adequate treatment options for distal femur fractures. Locked plating can be used for all distal femur fractures including complex type C fractures, periprosthetic fractures, as well as osteoporotic fractures. IM nailing provides favorable stability and can be successfully implanted in bilateral or multisegmental fractures of the lower extremity as well as in extra-articular fractures. However, both systems require precise preoperative planning and advanced surgical experience to reduce the risk of revision surgery. Clinical outcome largely depends on surgical technique rather than on the choice of implant.

Targeted disruption of LIGHT causes defects in costimulatory T cell activation and reveals cooperation with lymphotoxin beta in mesenteric lymph node genesis.

The recently described tumor necrosis factor (TNF) family member LIGHT (herpes virus entry mediator [HVEM]-L/TNFSF14), a ligand for the lymphotoxin (LT)beta receptor, HVEM, and DcR3, was inactivated in the mouse. In contrast to mice deficient in any other member of the LT core family, LIGHT(-/-) mice develop intact lymphoid organs. Interestingly, a lower percentage of LIGHT(-/-)LTbeta(-/-) animals contain mesenteric lymph nodes as compared with LTbeta(-/-) mice, whereas the splenic microarchitecture of LIGHT(-/-)LTbeta(-/-) and LTbeta(-/-) mice shows a comparable state of disruption. This suggests the existance of an additional undiscovered ligand for the LTbeta receptor (LTbetaR) or a weak LTalpha(3)-LTbetaR interaction in vivo involved in the formation of secondary lymphoid organs. LIGHT acts synergistically with CD28 in skin allograft rejection in vivo. The underlying mechanism was identified in in vitro allogeneic MLR studies, showing a reduced cytotoxic T lymphocyte activity and cytokine production. Detailed analyses revealed that proliferative responses specifically of CD8+ T cells are impaired and interleukin 2 secretion of CD4+ T cells is defective in the absence of LIGHT. Furthermore, a reduced 3[H]-thymidine incorporation after T cell receptor stimulation was observed. This for the first time provides in vivo evidence for a cooperative role for LIGHT and LTbeta in lymphoid organogenesis and indicates important costimulatory functions for LIGHT in T cell activation.