Evaluation of the influence of platelet-rich plasma (PRP), platelet lysate (PL) and mechanical loading on chondrogenesis in vitro.

The aim of this work is to investigate the capability of PRP as an adjuvant therapy to autologous chondrocyte implantation (ACI) in combination with multi-axial load with respect to cartilage regeneration. Articular cartilage shows poor repair capacity and therapies for cartilage defects are still lacking. Well-established operative treatments include ACI, and growing evidence shows the beneficial effects of PRP. Platelets contain numerous growth factors, among them transforming growth factor beta (TGF-ß). Dynamic mechanical loading is known to be essential for tissue formation, improving extracellular matrix (ECM) production. For our ACI model monolayer expanded human chondrocytes were seeded into polyurethane scaffolds and embedded in fibrin (hChondro), in PRP-Gel (PRP), or in fibrin with platelet lysate (PL), which was added to the media once a week with a concentration of 50 vol%. The groups were either exposed to static conditions or multi-axial forces in a ball-joint bioreactor for 1 h per day over 2 weeks, mimicking ACI under physiological load. The culture medium was collected and analyzed for glycosaminoglycan (GAG), nitrite and transforming growth factor beta 1 (TGF-ß1) content. The cell-scaffold constructs were collected for DNA and GAG quantification; the expression of chondrogenic genes, TGF-ß and related receptors, as well as inflammatory genes, were analyzed using qPCR. Loading conditions showed superior chondrogenic differentiation (upregulation of COL2A1, ACAN, COMP and PRG4 expression) than static conditions. PRP and PL groups combined with mechanical loading showed upregulation of COL2A1, ACAN and COMP. The highest amount of total TGF-ß1 was quantified in the PL group. Latent TGF-ß1 was activated in all loaded groups, while the highest amount was found in the PL group. Load increased TGFBR1/TGFBR2 mRNA ratio, with further increases in response to supplements. In general, loading increased nitrite release into the media. However, over time, the media nitrite content was lower in the PL group compared to the control group. Based on these experiments, we conclude that chondrogenic differentiation is strongest when simulated ACI is performed in combination with dynamic mechanical loading and PRP-gel or PL supplementation. An inflammatory reaction was reduced by PRP and PL, which could be one of the major therapeutic effects. Loading presumably can enhance the action of TGF-ß1, which was predominantly activated in loaded PL groups. The combination of load and PRP represents an effective and promising synergy concerning chondrocyte-based cartilage repair.

Vertebral osteomyelitis in patients with Staphylococcus aureus bloodstream infection: Evaluation of risk factors for treatment failure.

OBJECTIVES: Staphylococcus aureus is the most common cause of pyogenic vertebral osteomyelitis (VO). Studies indicate that S. aureus VO results in poor outcome. We aimed to investigate risk factors for treatment failure in patients with Staphylococcus aureus bloodstream infection (SAB) and VO. METHODS: We conducted a post hoc-analysis of data from a German bi-center prospective SAB cohort (2006-2014). Patients were followed-up for one year. Primary outcome was treatment failure defined as relapse and/or death within one year. RESULTS: A total of 1069 patients with SAB were analyzed, with 92 VO patients. In addition to antibiotic treatment, surgery was performed in 60/92 patients. Treatment failed in 44/92 patients (death, n = 42; relapse, n = 2). Multivariable analysis revealed higher age (HR 1.04 [per year], 95%CI 1.01-1.07), Charlson comorbidity index (HR 1.20, 95%CI 1.06-1.36), presence of neurologic deficits (HR 2.53, 95%CI 1.15-5.53) and local abscess formation (HR 3.35, 95%CI 1.39-8.04) as independent risk factors for treatment failure. In contrast, surgery seemed to be associated with a favourable outcome (HR 0.45 (95%CI 0.20-0.997)). CONCLUSION: SAB patients with VO exhibit a high treatment failure rate. Red flags are older age, comorbidities, neurologic deficits and local abscess formation. Whether these patients benefit from intensified treatment (e.g. radical surgery, prolongation of antibiotics) should be investigated further.

The tissue-renin-angiotensin-system of the human intervertebral disc.

Symptomatic intervertebral disc (IVD) degeneration accounts for significant socioeconomic burden. Recently, the expression of the tissue renin-angiotensin system (tRAS) in rat and bovine IVD was demonstrated. The major effector of tRAS is angiotensin II (AngII), which participates in proinflammatory pathways. The present study investigated the expression of tRAS in human IVDs, and the correlation between tRAS, inflammation and IVD degeneration. Human IVD tissue was collected during spine surgery and distributed according to principal diagnosis. Gene expression of tRAS components, proinflammatory and catabolic markers in the IVD tissue was assessed. Hydroxyproline (OHP) and glycosaminoglycan (GAG) content in the IVD tissue were determined. Tissue distribution of tRAS components was investigated by immunohistochemistry. Gene expression of tRAS components such as angiotensin-converting enzyme (ACE), Ang II receptor type 2 (AGTR2), angiotensinogen (AGT) and cathepsin D (CTSD) was confirmed in human IVDs. IVD samples that expressed tRAS components (n = 21) revealed significantly higher expression levels of interleukin 6 (IL-6), tumour necrosis factor α (TNF-α), a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) 4 and 5 compared to tRAS-negative samples (n = 37). Within tRAS-positive samples, AGT, matrix-metalloproteinases 13 and 3, IL-1, IL-6 and IL-8 were more highly expressed in traumatic compared to degenerated IVDs. Total GAG/DNA content of non-tRAS expressing IVD tissue was significantly higher compared to tRAS positive tissue. Immunohistochemistry confirmed the presence of AngII in the human IVD. The present study identified the existence of tRAS in the human IVD and suggested a correlation between tRAS expression, inflammation and ultimately IVD degeneration.

Biomechanical investigation of a minimally invasive posterior spine stabilization system in comparison to the Universal Spinal System (USS).

BACKGROUND: Although minimally invasive posterior spine implant systems have been introduced, clinical studies reported on reduced quality of spinal column realignment due to correction loss. The aim of this study was to compare biomechanically two minimally invasive spine stabilization systems versus the Universal Spine Stabilization system (USS). METHODS: Three groups with 5 specimens each and 2 foam bars per specimen were instrumented with USS (Group 1) or a minimally invasive posterior spine stabilization system with either polyaxial (Group 2) or monoaxial (Group 3) screws. Mechanical testing was performed under quasi-static ramp loading in axial compression and torsion, followed by destructive cyclic loading run under axial compression at constant amplitude and then with progressively increasing amplitude until construct failure. Bending construct stiffness, torsional stiffness and cycles to failure were investigated. RESULTS: Initial bending stiffness was highest in Group 3, followed by Group 2 and Group 1, without any significant differences between the groups. A significant increase in bending stiffness after 20'000 cycles was observed in Group 1 (p = 0.002) and Group 2 (p = 0.001), but not in Group 3, though the secondary bending stiffness showed no significant differences between the groups. Initial and secondary torsional stiffness was highest in Group 1, followed by Group 3 and Group 2, with significant differences between all groups (p ≤ 0.047). A significant increase in initial torsional stiffness after 20'000 cycles was observed in Group 2 (p = 0.017) and 3 (p = 0.013), but not in Group 1. The highest number of cycles to failure was detected in Group 1, followed by Group 3 and Group 2. This parameter was significantly different between Group 1 and Group 2 (p = 0.001), between Group 2 and Group 3 (p = 0.002), but not between Group 1 and Group 3. CONCLUSIONS: These findings quantify the correction loss for minimally invasive spine implant systems and imply that unstable spine fractures might benefit from stabilization with conventional implants like the USS.

Relationship between autologous bone graft osteointegration and correction loss after antero-posterior spondylodesis of traumatic vertebral body fracture.

BACKGROUND: A common method to restore the sagittal alignment and stabilize the spinal column is a dorso-ventral spondylodesis. It is assumed that correction loss after posttraumatic spondylodesis results from inadequate incorporation of the autologous iliac crest graft. MATERIALS AND METHODS: Retrospective documentation of patients with unstable vertebral body fractures of the thoracic or lumbar spine with concomitant rupture of at least one adjacent intervertebral disk who received surgical treatment at our institution from 2000 to 2006. Followed by analysis of the computer tomography documentation of a total of 142 patients with unstable vertebral body fracture stabilized by posterior internal fixator and anterior iliac crest spondylodesis. RESULTS: The following mean angle changes were derived from the second series of CT scans performed on average 283 days after anterior spondylodesis: vertebral wedge angle (VWA): 2.1°; segmental kyphotic angle: 4.9°; adjusted-SKA: 4.8°; sagittal index (SI): -0.04; segmental-scoliotic-angle (SSA): 0°; adjusted-SSA: 0°. Changes in VWA, both SKAs and SI postoperatively and prior to ME, were statistically significant (P<0.05). The McAfee fusion assessment of the graft showed: full fusion: cranial 64%, caudal 47%; partial fusion: cranial 20.5%, caudal 29%; lysis: cranial 8.5%, caudal 17%; graft resorption: 7%. No correlation was found between the above-mentioned angle changes and fusions grade. DISCUSSION: The importance of radiological evidence of fusion deficiency is questionable, because the extent of fusion only has a minimal effect on correction loss. LEVEL OF EVIDENCE: Level IV.

Distal tibial fractures and pilon fractures.

Fractures of the distal tibia are often very severe injuries that generally occur in the context of high-energy trauma and present with significant concomitant soft tissue involvement. Open fractures and extensive destruction of the articular surfaces are important challenges to the treating surgeon. In consequence the outcome for distal meta- and epiphyseal tibial fractures depends largely on the severity of the soft tissue injury and its management. Conventionally, tibial pilon fractures require surgical intervention. Conservative treatment would only be considered in some exceptional cases, for example, inoperability of the patient. Controversial discussion of optimal surgical technique and optimal timing of surgery is ongoing. There is broad consensus that soft tissue consolidation must have first priority as this is the basis for both fracture healing and good long-term outcomes. Surgical intervention can be managed as a one-stage or multi-stage procedure to achieve internal or external fracture fixation.

Anatomical assessment of iliac crest graft size for anterior spondylodesis.

Standard procedure for monosegmental anterior spondylodesis often combines anterior stabilization with autologous iliac crest graft. Recent publications defined a minimum size of the graft as a technical specification for this procedure. The cross sectional area of the graft to be transplanted should be at least 23.9% of the cross sectional area of the vertebral bodies to be fused. We investigated whether the required minimum size of autologous graft, as identified both experimentally and clinically, is compatible with the anatomical conditions in central european patients. Computed tomography scans (n = 348) of polytraumatized patients were obtained in the course of initial diagnosis. The scans were evaluated for vertebral body size and the possible size of autologous bone graft in the region of the anterior superior iliac crest. The evaluation of 348 CT scans demonstrated that 95% of the quantified iliac crest grafts would achieve the size recommended for anterior spinal fusion between T10 and T12. In 90% of all cases the planned iliac crest graft exceeded the size limit of 23,9% between concerning the endplates T10 and L2. In 85% the planned iliac crest graft exceeded the size limit of 23,9% between T10 and L3. The recommendation to take this value into account for monosegmental anterior spondylodesis should gain in importance in clinical practice.

Biomechanical comparison of three types of bone graft for anterior spondylodesis.

BACKGROUND: Anterior spondylodesis with bone grafting for fusion of spinal motion segments is a common procedure in clinical routine. Bone grafts for fusion include autologous, allogenic and xenogenic grafts that are inserted in the unstable spinal motion segment. Nevertheless, biomechanical data for autologous, allogenic and xenogenic grafts are rare. OBJECTIVE: The purpose of this study was to conduct biomechanical comparison of native and conserved bone grafts used in spondylodesis of the spine. METHODS: Grafts examined were native ovine tricortical iliac crest grafts, bovine cancellous bone blocks and ovine, tricortical iliac crest grafts, conserved in the same way as the bovine cancellous bone blocks. The grafts were tested biomechanically to failure. Compared parameters were maximum torque, maximum rotation angle and graft stiffness at failure. The Mann-Whitney-U test with Bonferroni adjustment was used for statistical analysis. RESULTS: Maximum torque at failure of the bovine cancellous bone graft did not differ significantly from that of the native ovine tricortical graft. Comparison of the conserved ovine tricortical graft revealed significantly lower values compared to the native ovine tricortical bone graft.Maximum rotation angle at failure of the untreated ovine tricortical bone grafts was significantly higher compared to the other grafts tested. Regarding graft stiffness no significant differences were found. CONCLUSIONS: Based on the functional demands exerted on the spinal motion segment, our results suggest that torsional strength and deformational behavior of the bone graft influence the stability of the spondylodesis. The native tricortical graft best fulfills this requirement.

[Radiological comparison between two procedures for ventral spondylodesis: autologous iliac crest bone graft vs bovine bone graft]. / Radiologischer Vergleich zweier Verfahren zur ventralen Spondylodese: Autologer trikortikaler Beckenkammspan vs. boviner Spongiosablock.

BACKGROUND: The objective of this study was the radiological evaluation of osseous integration of autologous iliac crest graft and bovine bone graft after spondylodesis based on a standardized score. MATERIAL AND METHODS: Spondylodesis was performed on 18 sheep, divided into 2 groups, 1 with an autologous iliac crest graft and the other with a bovine bone graft. Computed tomography was performed 12 and 24 weeks postoperatively. The osseous integration was assessed by the Tübinger Score. RESULTS: The evaluation of the CT scans demonstrated a significantly better osseous integration of the autologous iliac crest graft compared to the bovine bone graft. CONCLUSIONS: Based on our results, the bovine bone graft as a transplant for spondylodesis is inadvisable.

[Therapy of humeral shaft fractures]. / Therapie der Humerusschaftfraktur.

Fractures of the humeral shaft are less frequent than those of the proximal humerus. The formerly recommended treatment of humeral shaft fractures was conservative according to Böhler. This still remains an adequate concept of treatment but according to a change in the technical possibilities and the demands of patients and physicians on fast restoration of function and low pain, there is a trend towards surgical stabilization of humeral shaft fractures. The implant of choice is discussed controversially and consists of various types of nails versus plating. The technique of nailing is antegrade or retrograde and depends on the localization of the fracture. In our opinion good indications for plating are combined fractures of the proximal humerus and the shaft as well as very distal humeral shaft fractures. A primary lesion of the radial nerve is no imperative indication for exploration and different studies have shown the same results for exploration after 2 or 3 months if there is no spontaneous remission.

[Position Paper of the Youth Forum DGOU 2010 - part 2]. / Klausurtagung - Positionspapier Junges Forum der DGOU 2010 - Teil 2.

The aim of the "Youth Forum of the DGOU" during the Convention 2010 in Heidelberg was to place a statement concerning the professional politics in the field of Orthopaedic and Trauma Surgery. The Bologna Process realizes a standardization of the academic training within the European Union. For medicine this concept would raise the opportunity to opt out after three years with a bachelor degree applying for alternative occupations within the health care system. However, these alternative occupations are rarely defined and, in addition, the current structure of medical school in Germany provides the highest possible education for doctors in a direct and very well established way. Thus, reforming medical school in Germany into a Bachelor-master's system is an ambivalent approach, which considers a thorough reappraisal. There is currently no necessity for an speciality training in emergency medicine. The rapid and qualified treatment by the specific subspecialty provides a high standard of care for the patient. The high frequency exposure of the trauma and orthopaedic resident with emergency cases is an essential part of the current professional training.The "Junge Forum der DGOU" continues to understand the speciality training "Facharzt für Orthopädie und Unfallchirurgie" as the basic module of the profession. After that it should be possible to continuing subspecialty training and obtaining "spezielle Unfallchirurgie" bzw "spezielle orthopädische Chirurgie". After that further subspecialty training should be encouraged.

[Position Paper of the Youth Forum DGOU 2010 - part 1]. / Klausurtagung - Positionspapier Junges Forum der DGOU 2010 - Teil 1.

The aim of the "Youth Forum of the DGOU" during the Convention 2010 in Heidelberg was to place a statement concerning the professional politics in the field of Orthopaedic and Trauma Surgery. The emigration of young German physicians, the occupational image of the Physician Assistant and the quality of the German residency programs in Orthopaedic and Trauma Surgery we discussed as main topics. The main reason for young German physicians to go abroad is, besides better work conditions and less bureaucracy, the better structured education during residency. Therefore the "Youth Forum" generally supports the concept of the "physician assistant" as "a relief from non-physician duties rather than discussing the delegation of true physician duties". The "Youth Forum" is looking forward to collaborating on the improvement of the ongoing education of residents. In this regard, Orthopaedic and Trauma Surgery needs to become more attractive for young academics. We also support a uniform and nationwide curriculum, which guarantees a structured education to improve the theoretic, practical and academic skills of the future specialist in orthopaedic and trauma surgery. Additional surveys and interviews among the current generation of residents are needed to further specify the potential goals of such a curriculum. We would like to discuss the future of our speciality with our colleagues. Therefore different communication platforms including our website http://www.jf-dgou.de have been created.

[Graft integration in the lumbar spine of bovine cancellous bone compared to autologous iliac crest in a sheep model]. / Das Einwachsverhalten von trikortikalen Beckenkammspänen und lösungsmittelkonservierter, boviner Spongiosa bei ventraler Spondylodese der Lendenwirbelsäule im Schafsmodell.

BACKGROUND: Spinal injuries are common and a standard procedure for the stabilisation of spinal injuries is ventral spondylodesis with an autograft from the iliac crest. Because of the high incidence of harvesting complications there is a need to search for alternative materials. The aim of our study was to evaluate graft integration in the lumbar spine of bovine cancellous bone compared to autologous iliac crest material. MATERIAL AND METHODS: Two groups of eight female adult sheep (median age 3 years, range 2.4-3.8 years) received surgical treatment in the form of anterior monosegmental spondylodesis. The spondylodesis was performed in all animals in the motion segment L3/4 through a lateral approach with the animals lying on their right sides. To produce serial sections, the explanted vertebral segments were implanted in methyl methacrylate. On one side the histological preparation was examined qualitatively and in addition we analysed the quantity of the bone structure with special software. RESULTS: The bone structure in both groups did not differ significantly and demonstrated integration of the grafts in the adjacent vertebral bodies. Fractures and lysis occurred in the region of the intervertebral disc and were more frequent in the group with the bovine graft. CONCLUSION: The bony integration of the grafts of both groups was not significantly different and showed good results. Almost all of the bovine grafts fractured or presented regions of lysis. In our opinion bovine cancellous bone graft is not a good alternative to autologous iliac crest.