Wall teichoic acid substitution with glucose governs phage susceptibility of Staphylococcus epidermidis.

The species- and clone-specific susceptibility of Staphylococcus cells for bacteriophages is governed by the structures and glycosylation patterns of wall teichoic acid (WTA) glycopolymers. The glycosylation-dependent phage-WTA interactions in the opportunistic pathogen Staphylococcus epidermidis and in other coagulase-negative staphylococci (CoNS) have remained unknown. We report a new S. epidermidis WTA glycosyltransferase TagE whose deletion confers resistance to siphoviruses such as ΦE72 but enables binding of otherwise unbound podoviruses. S. epidermidis glycerolphosphate WTA was found to be modified with glucose in a tagE-dependent manner. TagE is encoded together with the enzymes PgcA and GtaB providing uridine diphosphate-activated glucose. ΦE72 transduced several other CoNS species encoding TagE homologs, suggesting that WTA glycosylation via TagE is a frequent trait among CoNS that permits interspecies horizontal gene transfer. Our study unravels a crucial mechanism of phage-Staphylococcus interaction and horizontal gene transfer, and it will help in the design of anti-staphylococcal phage therapies.IMPORTANCEPhages are highly specific for certain bacterial hosts, and some can transduce DNA even across species boundaries. How phages recognize cognate host cells remains incompletely understood. Phages infecting members of the genus Staphylococcus bind to wall teichoic acid (WTA) glycopolymers with highly variable structures and glycosylation patterns. How WTA is glycosylated in the opportunistic pathogen Staphylococcus epidermidis and in other coagulase-negative staphylococci (CoNS) species has remained unknown. We describe that S. epidermidis glycosylates its WTA backbone with glucose, and we identify a cluster of three genes responsible for glucose activation and transfer to WTA. Their inactivation strongly alters phage susceptibility patterns, yielding resistance to siphoviruses but susceptibility to podoviruses. Many different CoNS species with related glycosylation genes can exchange DNA via siphovirus ΦE72, suggesting that glucose-modified WTA is crucial for interspecies horizontal gene transfer. Our finding will help to develop antibacterial phage therapies and unravel routes of genetic exchange.

Invasive Staphylococcus epidermidis uses a unique processive wall teichoic acid glycosyltransferase to evade immune recognition.

Staphylococcus epidermidis expresses glycerol phosphate wall teichoic acid (WTA), but some health care-associated methicillin-resistant S. epidermidis (HA-MRSE) clones produce a second, ribitol phosphate (RboP) WTA, resembling that of the aggressive pathogen Staphylococcus aureus. RboP-WTA promotes HA-MRSE persistence and virulence in bloodstream infections. We report here that the TarM enzyme of HA-MRSE [TarM(Se)] glycosylates RboP-WTA with glucose, instead of N-acetylglucosamine (GlcNAc) by TarM(Sa) in S. aureus. Replacement of GlcNAc with glucose in RboP-WTA impairs HA-MRSE detection by human immunoglobulin G, which may contribute to the immune-evasion capacities of many invasive S. epidermidis. Crystal structures of complexes with uridine diphosphate glucose (UDP-glucose), and with UDP and glycosylated poly(RboP), reveal the binding mode and glycosylation mechanism of this enzyme and explain why TarM(Se) and TarM(Sa) link different sugars to poly(RboP). These structural data provide evidence that TarM(Se) is a processive WTA glycosyltransferase. Our study will support the targeted inhibition of TarM enzymes, and the development of RboP-WTA targeting vaccines and phage therapies.

UmetaFlow: an untargeted metabolomics workflow for high-throughput data processing and analysis.

Metabolomics experiments generate highly complex datasets, which are time and work-intensive, sometimes even error-prone if inspected manually. Therefore, new methods for automated, fast, reproducible, and accurate data processing and dereplication are required. Here, we present UmetaFlow, a computational workflow for untargeted metabolomics that combines algorithms for data pre-processing, spectral matching, molecular formula and structural predictions, and an integration to the GNPS workflows Feature-Based Molecular Networking and Ion Identity Molecular Networking for downstream analysis. UmetaFlow is implemented as a Snakemake workflow, making it easy to use, scalable, and reproducible. For more interactive computing, visualization, as well as development, the workflow is also implemented in Jupyter notebooks using the Python programming language and a set of Python bindings to the OpenMS algorithms (pyOpenMS). Finally, UmetaFlow is also offered as a web-based Graphical User Interface for parameter optimization and processing of smaller-sized datasets. UmetaFlow was validated with in-house LC-MS/MS datasets of actinomycetes producing known secondary metabolites, as well as commercial standards, and it detected all expected features and accurately annotated 76% of the molecular formulas and 65% of the structures. As a more generic validation, the publicly available MTBLS733 and MTBLS736 datasets were used for benchmarking, and UmetaFlow detected more than 90% of all ground truth features and performed exceptionally well in quantification and discriminating marker selection. We anticipate that UmetaFlow will provide a useful platform for the interpretation of large metabolomics datasets.

The influence of total disc arthroplasty with Mobidisc prosthesis on lumbar spine and pelvic parameters: a prospective in vivo biomechanical study with a minimum 3 year of follow-up.

BACKGROUND: This study examined the impact of Mobidisc implant on spinopelvic parameters, with particular focus on the preservation of the lumbar lordosis (LL) and on the segmental lordosis (SL) of the treated and adjacent segments. METHODS: A prospective study was conducted on 63 consecutive patients with symptomatic degenerative disc disease who underwent Mobidisc implantation at the Clinic for Spinal Diseases in Strasbourg, France. Based on the profile images of the whole, the following static spinopelvic parameters were measured and analysed: lumbar lordosis L1-S1 (LL), SL for L3-L4, L4-L5 and L5-S1, sacral slope (SS), pelvic tilt (PT) and pelvic incidence. In the lumbar spine images, the anterior (ADH) and posterior disc height (PDH) were measured prior to surgery and at the different follow-up appointments. The preoperative and postoperative values were compared and statistically analysed at different time intervals. RESULTS: Sixty-three patients were included in the study. The average age of the patients was 41.4 years (range 27-59 years). The mean follow-up was 44 months (range 36-71 months). Overall, total disc replacement (TDR) led to an increase in LL which increased TED over time. The preoperative LL measured 48.9° ± 10.1° and 53.4° ± 9.9° at 3 years follow-up (p < 0.0001). In the cohort of patients who underwent TDR at L4-5, the LL increased from 51.6° ± 10° to 56.2° ± 9.2° at the last FU (p = 0.006). All other spinopelvic parameters remained stable between the preoperative values and the last follow-up. In the patients who underwent L5-S1 TDR, a significant increase in LL was also observed between preoperative data and at the last FU (from 47.8° ± 10.1° to 53.3° ± 10.1°, p < 0.0001). Following L5-S1 TDR, the SS increased from 32.9° ± 8.3° to 35.6° ± 7.4° (p = 0.05) and the PT decreased from 15.4° ± 6.2° to 11.6° ± 5.7° between preoperative values and the last follow-up. Considering the entire cohort, the SL L5-S1 increased significantly from 5.9° ± 4.2° preoperatively to 8.1° ± 4.4° (p < 0.01) at the last FU, while at the L4-L5 level, the SL remained stable from 9.9 ± 4.5° to 10.7° ± 3.8° (p = 0.3). After L4-5 TDR, an increase in ADH and PDH at the treated level was observed, while these parameters progressively decreased in the adjacent segment. In patients who underwent L5-S1 TDR, a significant increase in L5-S1 ADH and PDH was observed from 18.8 ± 9.1 to 28.4 ± 11.1 and from 9.5 ± 3.8 to 17.6 ± 9.5 pixels, respectively. ADH and PDH at the proximal adjacent levels L3-4 and L4-5 were reduced. We did not observe any case of implant failure or damage to the bone/implant interface. CONCLUSION: TDR with Mobidisc allows for an improvement of LL and SL at the treated level. An increase in both anterior and posterior disc height was observed at the treated level. While disc height decreased at the adjacent level, further studies are required to investigate whether these changes are clinically relevant.

Staphylococcus epidermidis clones express Staphylococcus aureus-type wall teichoic acid to shift from a commensal to pathogen lifestyle.

Most clonal lineages of Staphylococcus epidermidis are commensals present on human skin and in the nose. However, some globally spreading healthcare-associated and methicillin-resistant S. epidermidis (HA-MRSE) clones are major causes of difficult-to-treat implant or bloodstream infections. The molecular determinants that alter the lifestyle of S. epidermidis have remained elusive, and their identification might provide therapeutic targets. We reasoned that changes in surface-exposed wall teichoic acid (WTA) polymers of S. epidermidis, which potentially shape host interactions, may be linked to differences between colonization and infection abilities of different clones. We used a combined epidemiological and functional approach to show that while commensal clones express poly-glycerolphosphate WTA, S. epidermidis multilocus sequence type 23, which emerged in the past 15 years and is one of the main infection-causing HA-MRSE clones, contains an accessory genetic element, tarIJLM, that leads to the production of a second, Staphylococcus aureus-type WTA (poly-ribitolphosphate (RboP)). Production of RboP-WTA by S. epidermidis impaired in vivo colonization but augmented endothelial attachment and host mortality in a mouse sepsis model. tarIJLM was absent from commensal human sequence types but was found in several other HA-MRSE clones. Moreover, RboP-WTA enabled S. epidermidis to exchange DNA with S. aureus via siphovirus bacteriophages, thereby creating a possible route for the inter-species exchange of methicillin resistance, virulence and colonization factors. We conclude that tarIJLM alters the lifestyle of S. epidermidis from commensal to pathogenic and propose that RboP-WTA might be a robust target for preventive and therapeutic interventions against MRSE infections.

Chitin, Chitin Oligosaccharide, and Chitin Disaccharide Metabolism of Escherichia coli Revisited: Reassignment of the Roles of ChiA, ChbR, ChbF, and ChbG.

Escherichia coli is unable to grow on polymeric and oligomeric chitin, but grows on chitin disaccharide (GlcNAc-GlcNAc; N,N'-diacetylchitobiose) and chitin trisaccharide (GlcNAc-GlcNAc-GlcNAc; N,N',N''-triacetylchitotriose) via expression of the chb operon (chbBCARFG). The phosphotransferase system (PTS) transporter ChbBCA facilitates transport of both saccharides across the inner membrane and their concomitant phosphorylation at the non-reducing end, intracellularly yielding GlcNAc 6-phosphate-GlcNAc (GlcNAc6P-GlcNAc) and GlcNAc6P-GlcNAc-GlcNAc, respectively. We revisited the intracellular catabolism of the PTS products, thereby correcting the reported functions of the 6-phospho-glycosidase ChbF, the monodeacetylase ChbG, and the transcriptional regulator ChbR. Intracellular accumulation of glucosamine 6P-GlcNAc (GlcN6P-GlcNAc) and GlcN6P-GlcNAc-GlcNAc in a chbF mutant unraveled a role for ChbG as a monodeacetylase that removes the N-acetyl group at the non-reducing end. Consequently, GlcN6P- but not GlcNAc6P-containing saccharides likely function as coactivators of ChbR. Furthermore, ChbF removed the GlcN6P from the non-reducing terminus of the former saccharides, thereby degrading the inducers of the chb operon and facilitating growth on the saccharides. Consequently, ChbF was unable to hydrolyze GlcNAc6P-residues from the non-reducing end, contrary to previous assumptions but in agreement with structural modeling data and with the unusual catalytic mechanism of the family 4 of glycosidases, to which ChbF belongs. We also refuted the assumption that ChiA is a bifunctional endochitinase/lysozyme ChiA, and show that it is unable to degrade peptidoglycans but acts as a bona fide chitinase in vitro and in vivo, enabling growth of E. coli on chitin oligosaccharides when ectopically expressed. Overall, this study revises our understanding of the chitin, chitin oligosaccharide, and chitin disaccharide metabolism of E. coli.

Phosphoglycerol-type wall and lipoteichoic acids are enantiomeric polymers differentiated by the stereospecific glycerophosphodiesterase GlpQ.

The cell envelope of Gram-positive bacteria generally comprises two types of polyanionic polymers linked to either peptidoglycan (wall teichoic acids; WTA) or to membrane glycolipids (lipoteichoic acids; LTA). In some bacteria, including Bacillus subtilis strain 168, both WTA and LTA are glycerolphosphate polymers yet are synthesized through different pathways and have distinct but incompletely understood morphogenetic functions during cell elongation and division. We show here that the exolytic sn-glycerol-3-phosphodiesterase GlpQ can discriminate between B. subtilis WTA and LTA. GlpQ completely degraded unsubstituted WTA, which lacks substituents at the glycerol residues, by sequentially removing glycerolphosphates from the free end of the polymer up to the peptidoglycan linker. In contrast, GlpQ could not degrade unsubstituted LTA unless it was partially precleaved, allowing access of GlpQ to the other end of the polymer, which, in the intact molecule, is protected by a connection to the lipid anchor. Differences in stereochemistry between WTA and LTA have been suggested previously on the basis of differences in their biosynthetic precursors and chemical degradation products. The differential cleavage of WTA and LTA by GlpQ reported here represents the first direct evidence that they are enantiomeric polymers: WTA is made of sn-glycerol-3-phosphate, and LTA is made of sn-glycerol-1-phosphate. Their distinct stereochemistries reflect the dissimilar physiological and immunogenic properties of WTA and LTA. It also enables differential degradation of the two polymers within the same envelope compartment in vivo, particularly under phosphate-limiting conditions, when B. subtilis specifically degrades WTA and replaces it with phosphate-free teichuronic acids.

Bacteria's different ways to recycle their own cell wall.

The ability to recover components of their own cell wall is a common feature of bacteria. This was initially recognized in the Gram-negative bacterium Escherichia coli, which recycles about half of the peptidoglycan of its cell wall during one cell doubling. Moreover, E. coli was shown to grow on peptidoglycan components provided as nutrients. A distinguished recycling enzyme of E. coli required for both, recovery of the cell wall sugar N-acetylmuramic acid (MurNAc) of the own cell wall and for growth on external MurNAc, is the MurNAc 6-phosphate (MurNAc 6P) lactyl ether hydrolase MurQ. We revealed however, that most Gram-negative bacteria lack a murQ ortholog and instead harbor a pathway, absent in E. coli, that channels MurNAc directly to peptidoglycan biosynthesis. This "anabolic recycling pathway" bypasses the initial steps of peptidoglycan de novo synthesis, including the target of the antibiotic fosfomycin, thus providing intrinsic resistance to the antibiotic. The Gram-negative oral pathogen Tannerella forsythia is auxotrophic for MurNAc and apparently depends on the anabolic recycling pathway to synthesize its own cell wall by scavenging cell wall debris of other bacteria. In contrast, Gram-positive bacteria lack the anabolic recycling genes, but mostly contain one or two murQ orthologs. Quantification of MurNAc 6P accumulation in murQ mutant cells by mass spectrometry allowed us to demonstrate for the first time that Gram-positive bacteria do recycle their own peptidoglycan. This had been questioned earlier, since peptidoglycan turnover products accumulate in the spent media of Gram-positives. We showed, that these fragments are recovered during nutrient limitation, which prolongs starvation survival of Bacillus subtilis and Staphylococcus aureus. Peptidoglycan recycling in these bacteria however differs, as the cell wall is either cleaved exhaustively and monosaccharide building blocks are taken up (B. subtilis) or disaccharides are released and recycled involving a novel phosphomuramidase (MupG; S.aureus). In B. subtilis also the teichoic acids, covalently bound to the peptidoglycan (wall teichoic acids; WTAs), are recycled. During phosphate limitation, the sn-glycerol-3-phosphate phosphodiesterase GlpQ specifically degrades WTAs of B. subtilis. In S. aureus, in contrast, GlpQ is used to scavenge external teichoic acid sources. Thus, although bacteria generally recover their own cell wall, they apparently apply distinct strategies for breakdown and reutilization of cell wall fragments. This review summarizes our work on this topic funded between 2011 and 2019 by the DFG within the collaborative research center SFB766.

Osteoarthritis and spontaneous fusion of facet joints after percutaneous instrumentation in thoracolumbar fractures.

PURPOSE: This retrospective study determined the rate of osteoarthritis and spontaneous facet joint fusion and analyzed risk factors related to patient characteristics, fracture type or surgical technique on pre- and postoperative CT after percutaneous instrumentation in thoracolumbar fractures. METHODS: 1050 facet joints adjacent to screws in 148 patients (15-85 years) with thoracolumbar fractures were analyzed with an average time between CTs of 12.3 months. Screw diameters, lengths and cement augmentation were recorded. Facet joint violation by screw trajectory and by insertion depth was classified in three grades. Pre- and postoperative osteoarthritis was graded as absent, minor or severe and postoperative facet joint fusion as absent, partial or complete. RESULTS: The facet violation rate was moderate in 15.4% and severe in 0.6% according to screw trajectory, and 11.0 and 0.6%, respectively, according to insertion depth. Osteoarthritis was preoperatively rated moderate in 9.6% and severe in 1.2%. A progression was evidenced in 79 facet joints (7.5%). Screw cement augmentation was the main predictive factor (p < 0.0001). Partial fusion was evidenced in 2.6% and complete fusion in 1% of facet joints. Risk factors were: BMI (p = 0.0002), age (p = 0.0013), preoperative osteoarthritis (p = 0.0005), time between 2 CTs (p = 0.0001), B-type fractures (p = 0.0005), concomitant anterior fusion (p = 0.0034). CONCLUSIONS: Occurrence or worsening of osteoarthritis was mainly observed in elderly patients with cement-augmented screws and spontaneous facet fusion in elderly patients with high BMI and preoperative osteoarthritis, or in anteriorly fused B-type injuries. Thus, percutaneous instrumentation can safely be removed after fracture consolidation in younger patients while preserving facet joints.

Open versus percutaneous instrumentation in thoracolumbar fractures: magnetic resonance imaging comparison of paravertebral muscles after implant removal.

OBJECTIVE Percutaneous instrumentation in thoracolumbar fractures is intended to decrease paravertebral muscle damage by avoiding dissection. The aim of this study was to compare muscles at instrumented levels in patients who were treated by open or percutaneous surgery. METHODS Twenty-seven patients underwent open instrumentation, and 65 were treated percutaneously. A standardized MRI protocol using axial T1-weighted sequences was performed at a minimum 1-year follow-up after implant removal. Two independent observers measured cross-sectional areas (CSAs, in cm2) and region of interest (ROI) signal intensity (in pixels) of paravertebral muscles by using OsiriX at the fracture level, and at cranial and caudal instrumented pedicle levels. An interobserver comparison was made using the Bland-Altman method. Reference ROI muscle was assessed in the psoas and ROI fat subcutaneously. The ratio ROI-CSA/ROI-fat was compared for patients treated with open versus percutaneous procedures by using a linear mixed model. A linear regression analyzed additional factors: age, sex, body mass index (BMI), Pfirrmann grade of adjacent discs, and duration of instrumentation in situ. RESULTS The interobserver agreement was good for all CSAs. The average CSA for the entire spine was 15.7 cm2 in the open surgery group and 18.5 cm2 in the percutaneous group (p = 0.0234). The average ROI-fat and ROI-muscle signal intensities were comparable: 497.1 versus 483.9 pixels for ROI-fat and 120.4 versus 111.7 pixels for ROI-muscle in open versus percutaneous groups. The ROI-CSA varied between 154 and 226 for open, and between 154 and 195 for percutaneous procedures, depending on instrumented levels. A significant difference of the ROI-CSA/ROI-fat ratio (0.4 vs 0.3) was present at fracture levels T12-L1 (p = 0.0329) and at adjacent cranial (p = 0.0139) and caudal (p = 0.0100) instrumented levels. Differences were not significant at thoracic levels. When adjusting based on age, BMI, and Pfirrmann grade, a significant difference between open and percutaneous procedures regarding the ROI-CSA/ROI-fat ratio was present in the lumbar spine (p < 0.01). Sex and duration of instrumentation had no significant influence. CONCLUSIONS Percutaneous instrumentation decreased muscle atrophy compared with open surgery. The MRI signal differences for T-12 and L-1 fractures indicated less fat infiltration within CSAs in patients who received percutaneous treatment. Differences were not evidenced at thoracic levels, where CSAs were smaller. Fat infiltration was not significantly different at lumbar levels with either procedure in elderly patients with associated discopathy and higher BMI. In younger patients, there was less fat infiltration of lumbar paravertebral muscles with percutaneous procedures.

Temporary Percutaneous Instrumentation and Selective Anterior Fusion for Thoracolumbar Fractures.

STUDY DESIGN: Prospective clinical trial in thoracolumbar trauma with 5-year follow-up. OBJECTIVE: To analyze clinical and radiographic outcomes of minimal invasive surgery, and the rational of circumferential fracture treatment with regard to age, degenerative changes, bone mineral density, and global sagittal balance. SUMMARY OF BACKGROUND DATA: Non-neurologic fractures with anterior column defect can be treated by posterior percutaneous instrumentation and selective anterior fusion. After consolidation, instrumentation can be removed at 1 year to provide mobility in non-fused segments. METHODS: Fifty-one patients, 47 (18-75) years, were operated for A2, A3, or B-type fractures. Visual analog scale (VAS) for back pain and Oswestry Disability Index (ODI) were assessed. Radiographic measurements were: sagittal index, regional kyphosis, T4-T12 kyphosis, L1-S1 lordosis, pelvic incidence, pelvic tilt, sacral slope, and T9 tilt. Anterior fusion and facet joints were analyzed on computed tomography (CT) at 1 year. RESULTS: The ODI was 8.8 before accident, 35.4 at 3 months, 17.8 at 2 years, 14.4 at 5 years. The VAS was 2.0 at 3 months and 1.0 at 5 years. The sagittal index was 18.0° preoperatively and 1.0° at 3 months (P < 0.0001). A loss of reduction of 1.1° occurred after implant removal (P = 0.009). Global sagittal balance remained unchanged. Ten patients with osteopenia or osteoporosis had a worse ODI: 24.7 versus 11.9 (P = 0.016), and a greater loss of correction: 4.9° versus 1.3° (P = 0.007). Cages filled with cancellous bone from the fractured vertebra fused regularly. Spontaneous facet joint fusions were observed in two patients at the fracture level in B-type injuries. CONCLUSION: Percutaneous instrumentation and selective anterior fusion using autologous bone and mesh cages lead to high fusion rates, which provided good long-term clinical results in younger patients with thoracolumbar fractures. Sagittal alignment was maintained after instrumentation removal without damaging paravertebral muscles. Outcomes were worse in elderly patients presenting osteopenia or osteoporosis. LEVEL OF EVIDENCE: 3.

Surgical-site infection in spinal injury: incidence and risk factors in a prospective cohort of 518 patients.

PURPOSE: To investigate the incidence of surgical-site infection (SSI) and determinate the risk factors of SSI in the context of spinal injury. METHODS: From February 1, 2011 to July 31, 2011, for a multicentre cohort of patients with acute spinal injury, we prospectively censored those with SSI for at least 12 months. We recorded epidemiologic characteristics and details of surgical procedure and postoperative care for each patient. We calculated the incidence of SSI at 1, 3 and 12 months after surgery. Univariate and multivariate analysis were used to establish the association of risk factors and SSI. We studied clinical outcomes by a visual analog scale for pain and physical and mental component summaries (PCS and MCS) of the Medical Outcomes Survey 36-Item Short Form (SF-36). RESULTS: At 1 year, among 518 patients, we recorded 25 SSI events, with median occurrence at 16 days (25-75 % quartile: 13-44 days). Incidence of SSI was 3.2 % (95 % confidence interval [1.9-5.3 %]) at 1 month, 3.7 % (95 % [2.2-5.8 %]) at 3 months and 4.6 % (95 % CI [3-6.9 %]) at 12 months. On multivariate analysis, age, presence of diabetes and surgical duration were predictors of SSI (p = 0.009, p = 0.047, and p = 0.015 respectively). At 12 months, infected and non-infected patients did not differ in pain (p = 0.58) or SF-36 PCS (p = 0.8) or MCS (p = 0.68). CONCLUSIONS: In this large prospective multicentre study in the context of spinal injury, we obtained an equivalent incidence rate and risk factors of SSI as found in the literature for elective spinal surgery.

Fusion Rates of Different Anterior Grafts in Thoracolumbar Fractures.

STUDY DESIGN: Retrospective CT analysis of anterior fusion in thoracolumbar trauma. OBJECTIVE: The aim of this study was to compare fusion rates of different bone grafts and to analyze risk factors for pseudarthrosis. SUMMARY OF BACKGROUND DATA: Interbody fusion is indicated in anterior column defects. Different grafts are used: autologous iliac crest, titanium mesh cages filled with cancellous bone, and autologous ribs. It is not clear which graft offers the most reliable fusion. MATERIAL AND METHODS: Radiologic data of 116 patients (71 men, 45 women) operated for type A2, A3, B, or C fractures were analyzed. The average age was 44.6 years (range, 16-75 y) and follow-up was 2.7 years (range, 1-9 y). All patients were treated by posterior instrumentation followed by an anterior graft: 53 cases with iliac crest, 43 cases with mesh cages, and 20 with rib grafts. Fusion was evaluated on CT and classified into complete fusion, partial fusion, unipolar pseudarthrosis, and bipolar pseudarthrosis. RESULTS: Iliac crest fused in 66%, cages in 98%, and rib grafts in 90%. The fusion rate of cages filled with bone was significantly higher as the iliac graft fusion rate (P=0.002). The same was applied to rib grafts compared with iliac crest (P=0.041). Additional bone formation around the main graft, bridging both vertebral bodies, was observed in 31 of the 53 iliac crests grafts. Pseudarthrosis occurred more often in smokers (P=0.042). A relationship between fracture or instrumentation types, sex, age, BMI, and fusion could not be determined. CONCLUSIONS: Tricortical iliac crest grafts showed an unexpected high pseudarthrosis rate in thoracolumbar injuries. Their cortical bone is dense and their fusion surface is small. Rib grafts led to a better fusion when used in combination with the cancellous bone from the fractured vertebral body. Titanium mesh cages filled with cancellous bone led to the highest fusion rate and built a complete bony bridge between vertebral bodies. Smoking seemed to influence fusion. LEVEL OF EVIDENCE: Case control study, Level III.

Thoracolumbar fracture reduction by percutaneous in situ contouring.

PURPOSE: Percutaneous in situ contouring is based on bilateral bending of rods on the spine, thus increasing lordosis at the fracture. It was analyzed if this technique would provide a better reduction than prone positioning and how sagittal alignment would behave. METHODS: Twenty-nine patients were operated using in situ contouring and selective anterior fusion for non-neurologic A2, A3 or B2 fractures. Clinical results were assessed prospectively using visual analog scale (VAS) and Oswestry Disability Index (ODI). The radiographic deformity correction was measured by sagittal index and regional kyphosis. Sagittal balance was assessed using kyphosis, lordosis, T9 tilt, pelvic incidence, pelvic tilt and sacral slope. Posterior wall fragment reduction was evaluated by computed tomography. RESULTS: After 2 years, VAS and ODI were comparable to the status prior to the accident. The sagittal index was 19.7° preoperatively, 5.3° after prone positioning and -1.1° after in situ contouring (p < 0.001). The loss of correction was 2.4°, mainly during the first 3 months. Similar observations were made for regional kyphosis. The sagittal spino-pelvic alignment was stable postoperatively. A preoperative canal obstruction ≥50 % was observed in 16 patients, and the fragments migrated anteriorly in all patients. CONCLUSIONS: Percutaneous instrumentation and anterior fusion provides good clinical results. In situ contouring increases lordosis obtained by prone positioning. Anterior column lengthening and ligamentotaxis reduce posterior wall fragments, which decompress the canal without laminectomy. The fusion of anterior defects prevents the loss of correction and provides a stable sagittal profile. The instrumentation may be removed without damaging the paravertebral muscles and loss of correction.

Sagittal balance correction of idiopathic scoliosis using the in situ contouring technique.

PURPOSE: Idiopathic scoliosis can lead to sagittal imbalance. The relationship between thoracic hyper- and hypo-kyphotic segments, vertebral rotation and coronal curve was determined. The effect of segmental sagittal correction by in situ contouring was analyzed. METHODS: Pre- and post-operative radiographs of 54 scoliosis patients (Lenke 1 and 3) were analyzed at 8 years follow-up. Cobb angles and vertebral rotation were determined. Sagittal measurements were: kyphosis T4-T12, T4-T8 and T9-T12, lordosis L1-S1, T12-L2 and L3-S1, pelvic incidence, pelvic tilt, sacral slope, T1 and T9 tilt. RESULTS: Thoracic and lumbar curves were significantly reduced (p = 0.0001). Spino-pelvic parameters, T1 and T9 tilt were not modified. The global T4-T12 kyphosis decreased by 2.1° on average (p = 0.066). Segmental analysis evidenced a significant decrease of T4-T8 hyperkyphosis by 6.6° (p = 0.0001) and an increase of segmental hypokyphosis T9-T12 by 5.0° (p = 0.0001). Maximal vertebral rotation was located at T7, T8 or T9 and correlated (r = 0.422) with the cranial level of the hypokyphotic zone (p = 0.003). This vertebra or its adjacent levels corresponded to the coronal apex in 79.6 % of thoracic curves. CONCLUSIONS: Lenke 1 and 3 curves can show normal global kyphosis, divided in cranial hyperkyphosis and caudal hypokyphosis. The cranial end of hypokyphosis corresponds to maximal rotation. These vertebrae have most migrated anteriorly and laterally. The sagittal apex between segmental hypo- and hyper-kyphosis corresponds to the coronal thoracic apex. A segmental sagittal imbalance correction is achieved by in situ contouring. The concept of segmental imbalance is useful when determining the levels on which surgical detorsion may be focused.

Could an osteoinductor result in degeneration of a neurofibroma in NF1?

We report a case of fatal evolution of neurofibromatosis in a young boy. A laminectomy was performed when he was 9 years old. A secondary hyperkyphosis led to many surgeries resulting in recurrent malunions. When he was 23 years old, a breakage of his rods was treated by a new instrumentation and a T12-L1 interbody cage fitted with rh-BMP. Five months later, he developed a huge posterior tumour on his back. The biopsy diagnosed a neurofibrosarcoma. The growth of the tumour was extremely rapid. He died after several months from a septic shock. NF1 is characterised by neurofibromas that have a possibility of malign degeneration and conversion to a sarcoma. However, the chronology, rapidity of evolution and the exceptional volume of the tumour made us wonder whether the BMP had a part of responsibility as osteoinductor in the malignant degeneration, in this particular case, of neurofibromatosis. It seemed important to point out this case to the medical community.