Outcome of Conservative Therapy of Adolescent Idiopathic Scoliosis (AIS) with Chêneau-Brace.

Chêneau-brace (C-Brace) is a potential tool for the treatment of adolescent idiopathic scoliosis (AIS) with a Cobb angle between 20° and 45° for the primary curve. The aim of the present study was (1) to estimate study cohorts with C-brace therapy success and therapy failure and (2) to analyze possible factors that influence the therapy outcome. Seventy-eight patients with AIS were assessed before the initiation of C-brace treatment. Each patient underwent radiography examinations before the brace, in-brace, and at the therapy end. Cobb angle was considered as increased when the value at the end of therapy was increased more than 5° (Δ > 5°), unchanged-when the value was unchanged within ± 5° and decreased- when the value was decreased more than 5° (Δ < -5°). The study cohort was stratified due to curve topography in the thoracic, thoracolumbar, and lumbar scoliosis groups. Global analysis revealed no statistically significant modification of the Cobb angle (Cobb angle pre-brace vs. Cobb angle post-brace: 30.8° ± 8.2 vs. 29.3° ± 15.2, p = 0.26). However, at the end of C-brace therapy, the primary Cobb angle was decreased by more than 5° in 27 patients (35%), unchanged (Δ within the range of ±5°) in 36 patients (46%), and increased more than 5° in 15 patients (19%). Sub-group analysis due to curve topography and skeletal maturity has shown higher rates of brace therapy failure in thoracic curves and in younger patients (Risser grade 0). Patients with higher Cobb angle correction with C-brace had lower rates of therapy failure. The C-brace can be useful for the prevention of scoliotic curve progression in patients with AIS. However, many factors influence the therapy effect.

Return to Sport after Adolescent Idiopathic Scoliosis (AIS) Correction Surgery: A Retrospective Data Analysis.

Sports are relevant to younger populations in society. Adolescent idiopathic scoliosis (AIS) patients who undergo surgical correction of the spine are often intensively involved in sports. For that, returning to the sport is often an important concern for the patients and their families. To the best of our knowledge, there is still a lack of scientific data indicating established recommendations about the time of returning to sport activities after surgical spinal correction. The aim of this study was to investigate (1) when AIS patients return to athletic activities after a posterior fusion, and (2) if they change their activities postoperatively. Furthermore, another question was (3) if the length of the performed posterior fusion or (4) fusion to the lower lumbar spine could have an influence on the rates or time of returning to sport activities postoperatively. Data collection was performed using questionnaires assessing patients' contentment and athletic activity. Athletic activities were categorized into three categories: (1) contact, (2) contact/non-contact and (3) non-contact sports. The intensity of exercised sports, the time of returning to the sport and changes in sport habits were documented. Radiographs were evaluated pre- and postoperatively to determine the Cobb angle and the length of the posterior fusion via the identification of the upper (UIV) and lower instrumented vertebra (LIV). Stratification analysis due to the fusion length was performed to answer a hypothetical question. This retrospective survery of 113 AIS patients treated with a posterior fusion revealed that, on average, returning to sport activities required 8 months of postoperative rest. The preoperative to postoperative rate of patients participating in sport activities increased from 88 (78%) to 94 (89%). Furthermore, postoperatively, a relevant shift of exercised activities from contact to non-contact sports was noted. Further subanalysis revealed that only 33 subjects were able to return to exactly the same athletic activities as before surgery (10 months postoperatively). The assessment of radiographs revealed that in this study group, the length of the performed posterior fusion and fusions to the lower lumbar spine had no influence on the time of return to athletic activities. The results of this study might shed some light on postoperative recommendations for sport activities after AIS treatment with a posterior fusion and may be beneficial for surgeons treating patients.

[Anesthesia in spinal surgery]. / Anästhesie bei Wirbelsäulenoperationen.

Over the past 20 years improvements in surgical techniques and perioperative patient care have led to a considerable increase in surgical procedures of the spine worldwide. Therefore, the spectrum was extended from minimally invasive procedures up to complex operations over several segments of the spinal column with high loss of blood and complex perioperative management. This article presents the principal pillars of preoperative, intraoperative and postoperative management relating to spinal surgery. Furthermore, procedure-specific features, such as airway management in cervical spine instability or implementation of intraoperative neuromonitoring are dealt with in detail.

Influence of Chêneau-Brace Therapy on Lumbar and Thoracic Spine and Its Interdependency with Cervical Spine Alignment in Patients with Adolescent Idiopathic Scoliosis (AIS).

Chêneau-brace is an effective therapy tool for treatment in adolescent idiopathic scoliosis (AIS). Data on potential interdependent changes of the sagittal profile including the cervical spine are still sparse. The purpose of this study was to evaluate in-brace changes of the thoracic and lumbar spine and their influence on the pelvis and the cervical spine and apical vertebral rotation was reported. Ninety-three patients with AIS undergoing Chêneau-bracing were included. Patients were stratified by lumbar, thoracic and global spine alignment into normolordotic vs. hyperlordotic or normokyphotic vs. hypokyphotic or anteriorly aligned vs. posteriorly aligned groups. The coronal Cobb angle was significantly decreased in all groups indicating good correction while in-brace therapy. Sagittally, in-brace treatment led to significant flattening of lumbar lordosis (LL) in all stratified groups. Thoracic kyphosis (TK) was significantly flattened in the normokyphotic group, but no TK changes were noticed in the hypokyphotic group. Pelvic tilt (PT) stayed unchanged during the in-brace therapy. Chêneau-brace showed marginal changes in the lower cervical spine but had no influence on the upper cervical spine. The apical vertebral axis in primary and secondary curves was unchanged during the first radiological follow-up. Results from this study contribute to better understanding of initial spine behavior in sagittal and axial plane in the context of bracing.

Investigation of Risk Factors for Pain Chronification in Patients Suffering from Infections of the Spine.

BACKGROUND: Spinal infections represent a therapeutic challenge. The often protracted course of the disease is accompanied by pain, which can lead to a chronic pain experience even after the infectious disease has been treated successfully. The aim of this study was to investigate possible risk factors of pain chronification. METHODS: In a prospective study, 14 patients with spinal infections were examined at admission (T1), at discharge from inpatient therapy (T2), and three to eight months postoperatively (T3) byquestionnaires on risk factors for pain chronification and by quantitative sensory testing (QST). RESULTS: In-patient treatment lasted on average 45.3 days (±33.13). The patients complained of pain for 3.43 months (±2.77) prior to inpatient treatment. The visual analogue scale (VAS) for pain (0-10) and the Oswestry Disability Index detected significant improvement in the course of the study. However, patients also reported catastrophic thinking, as well as fear of movement and (re)-injury. CONCLUSION: In summary, our results demonstrate that patients with spinal infections did not suffer from pain chronification, but might benefit from an interdisciplinary therapeutic approach, which emphasizes promoting active pain-coping strategies, as well as addressing fear of movement and catastrophic thinking.

[Battle: Indication for surgery in hip-spine syndrome-Hip or spine first? : The spine surgeon's view]. / Battle: Operationsindikation beim Hip-Spine-Syndrom ­ Die Hüfte oder die Wirbelsäule zuerst? : Die Sicht des Wirbelsäulenchirurgen.

Pathologies of the hip, spine and the lower extremity are often concomitant due to their three-dimensional anatomic and physiological interrelation. The real challenge lies in defining which of the pathologies is most relevant for the patient in terms of clinical symptoms and which organ should be treated first. The purpose of this review article is two-fold: Firstly, to explain the treatment dilemma of hip-spine syndrome to the treating physician. Secondly, to highlight the significance of spinal pathology in this context.

Three-Dimensional Analysis of Initial Brace Correction in the Setting of Adolescent Idiopathic Scoliosis.

The three-dimensional nature of adolescent idiopathic scoliosis (AIS) necessitates a tridimensional assessment and management. Bracing constitutes the mainstay conservative treatment for mild adolescent idiopathic scoliosis. In the literature hitherto, there has been uncertainty regarding the behavior of the spine, pelvis, and vertebral orientations in the context of bracing, especially in the transverse plane. This poses a challenge to healthcare providers, patients, and their families, as brace treatment, although not as invasive as surgery, is laden with medical and psychological complications and could be considered traumatizing. Hence, a thorough understanding of initial three-dimensional spinal behavior in the context of bracing is important. The purpose of this retrospective study was to investigate the immediate 3D impact of Chêneau-type brace. Thirty-eight patients with AIS undergoing Chêneau-type bracing were included. Patients were stratified according to their structural curve topography into thoracic, thoracolumbar, and lumbar groups. 3D reconstruction of the spine using a dedicated biplanar stereoradiography software with and without the brace was performed. The examined anthropometric radiographic measures were pre- to in-brace variations and differences of spinopelvic parameters and vertebral orientations in the coronal, sagittal, and transverse planes. The complex impact of the Chêneau-type brace on different curves in three planes was delineated. In the coronal plane, the Cobb angle was significantly decreased in all types of curves, and the coronal tilt correction was concentrated in specific segments. The impact of the brace in this study on the sagittal profile was variable, including the loss of thoracic kyphosis and lumbar lordosis. In the transverse plane, an axial vertebral rotation change and detorsion above the apex occurred in the thoracolumbar curves. The results from this exploratory study could shed some light on the initial 3D spinal behavior in the context of bracing and may be of beneficial for treating physicians and brace makers.

Impact of Electrocautery on Fatigue Life of Spinal Fusion Constructs-An In Vitro Biomechanical Study.

Instrumentation failure in the context of spine surgery is attributed to cyclic loading leading to formation of fatigue cracks, which later propagate and result in rod fracture. A biomechanical analysis of the potential impact of electrocautery on the fatigue life of spinal implants has not been previously performed. The aim of this study was to assess the fatigue life of titanium (Ti) and cobalt-chrome (CoCr) rod-screw constructs after being treated with electrocautery. Twelve spinal constructs with CoCr and Ti rods were examined. Specimens were divided into four groups by rod material (Ti and CoCr) and application of monopolar electrocautery on the rods' surface (control-group and electrocautery-group). Electrocautery was applied on each rod at three locations, then constructs were cyclically tested. Outcome measures were load-to-failure, total number of cycles-to-failure, and location of rod failure. Ti-rods treated with electrocautery demonstrated a significantly decreased fatigue life compared to non-treated Ti-rods. Intergroup comparison of cycles-to-failure revealed a significant mean decrease of almost 9 × 105 cycles (p = 0.03). No CoCr-rods failed in this experiment. Electrocautery application on the surface of Ti-rods significantly reduces their fatigue life. Surgeons should exercise caution when using electrocautery in the vicinity of Ti-rods to mitigate the risk of rod failure.

Sagittal alignment of the cervical spine in the setting of adolescent idiopathic scoliosis.

OBJECTIVEThe goal of this study was to investigate the impact of thoracic and lumbar alignment on cervical alignment in patients with adolescent idiopathic scoliosis (AIS).METHODSEighty-one patients with AIS who had a Cobb angle > 40° and full-length spine radiographs were included. Radiographs were analyzed using dedicated software to measure pelvic parameters (sacral slope [SS], pelvic incidence [PI], pelvic tilt [PT]); regional parameters (C1 slope, C0-C2 angle, chin-brow vertical angle [CBVA], slope of line of sight [SLS], McRae slope, McGregor slope [MGS], C2-7 [cervical lordosis; CL], C2-7 sagittal vertical axis [SVA], C2-T3, C2-T3 SVA, C2-T1 Harrison measurement [C2-T1 Ha], T1 slope, thoracic kyphosis [TK], lumbar lordosis [LL], and PI-LL mismatch); and global parameters (SVA). Patients were stratified by their lumbar alignment into hyperlordotic (LL > 59.7°) and normolordotic (LL 39.3° to 59.7°) groups and also, based on their thoracic alignment, into hypokyphotic (TK < -33.1°) and normokyphotic (TK -33.1° to -54.9°) groups. Finally, they were grouped based on their global alignment into either an anterior-aligned group or a posterior-aligned group.RESULTSThe lumbar hyperlordotic group, in comparison to the normolordotic group, had a significantly larger LL, SS, PI (all p < 0.001), and TK (p = 0.014) and a significantly smaller PI-LL mismatch (p = 0.001). Lumbar lordosis had no influence on local cervical parameters.The thoracic hypokyphotic group had a significantly larger PI-LL mismatch (p < 0.002) and smaller T1 slope (p < 0.001), and was significantly more posteriorly aligned than the normokyphotic group (-15.02 ± 8.04 vs 13.54 ± 6.17 [mean ± SEM], p = 0.006). The patients with hypokyphotic AIS had a kyphotic cervical spine (cervical kyphosis [CK]) (p < 0.001). Furthermore, a posterior-aligned cervical spine in terms of C2-7 SVA (p < 0.006) and C2-T3 SVA (p < 0.001) was observed in the thoracic hypokyphotic group.Comparing patients in terms of global alignment, the posterior-aligned group had a significantly smaller T1 slope (p < 0.001), without any difference in terms of pelvic, lumbar, and thoracic parameters when compared to the anterior-aligned group. The posterior-aligned group also had a CK (-9.20 ± 1.91 vs 5.21 ± 2.95 [mean ± SEM], p < 0.001) and a more posterior-aligned cervical spine, as measured by C2-7 SVA (p = 0.003) and C2-T3 SVA (p < 0.001).CONCLUSIONSAlignment of the cervical spine is closely related to thoracic curvature and global alignment. In patients with AIS, a hypokyphotic thoracic alignment or posterior global alignment was associated with a global cervical kyphosis. Interestingly, upper cervical and cranial parameters were not statistically different in all investigated groups, meaning that the upper cervical spine was not recruited for compensation in order to maintain a horizontal gaze.

Surgical site infections following instrumented stabilization of the spine.

BACKGROUND: Implant-associated infections are still a feared complication in the field of orthopedics. Bacteria attach to the implant surface and form so-called biofilm colonies that are often difficult to diagnose and treat. Since the majority of studies focus on prosthetic joint infections (PJIs) of the hip and knee, current treatment options (eg, antibiotic prophylaxis) of implant-associated infections have mostly been adapted according to these results. OBJECTIVE: The aim of this study was to evaluate patients with surgical site infections following instrumented stabilization of the spine with regard to detected bacteria species and the course of the disease. PATIENTS AND METHODS: We performed a retrospective single-center analysis of implant-associated infections of the spine from 2010 to 2014. A total of 138 patients were included in the study. The following parameters were evaluated: C-reactive protein serum concentration, microbiological evaluation of tissue samples, the time course of the disease, indication for instrumented stabilization of the spine, localization of the infection, and the number of revision surgeries required until cessation of symptoms. RESULTS: Coagulase-negative Staphylococcus spp. were most commonly detected (n=69, 50%), followed by fecal bacteria (n=46, 33.3%). In 23.2% of cases, no bacteria were detected despite clinical suspicion of an infection. Most patients suffered from degenerative spine disorders (44.9%), followed by spinal fractures (23.9%), non-degenerative scoliosis (20.3%), and spinal tumors (10.1%). Surgical site infections occurred predominantly within 3 months (64.5%), late infections after 2 years were rare (4.3%), in particular when compared with PJIs. Most cases were successfully treated after 1 revision surgery (60.9%), but there were significant differences between bacteria species. Fecal bacteria were more difficult to treat and often required more than 1 revision surgery. CONCLUSION: In summary, we were able to demonstrate significant differences between spinal implant-associated infections and PJIs. These aspects should be considered early on in the treatment of surgical site infections following instrumented stabilization of the spine.

A systematic review of the clinical applications and complications of bone marrow aspirate concentrate in management of bone defects and nonunions.

PURPOSE: Fracture healing encompasses a succession of dynamic multifactorial metabolic events, which ultimately re-establishes the integrity of the biomechanical properties of the bone. Up to 10% of the fractures occurring annually will need additional surgical procedures because of impaired healing. The aim of this article is to review the current literature regarding the use of bone marrow aspirate concentrate (BMAC) and its effectiveness in the management of bone defects. METHODS: We have included all published clinical literature investigating the development, techniques and applications of BMAC. Language, design and risk of bias did not deter the initial inclusion of any study. Our search was exclusively limited to studies involving human subjects. A PRISMA compliant search was carried out as published in 2009. This included the online databases: PubMed, EMBASE, clinical trial.gov and the Cochrane library from 1960 to the end of May 2015. MeSH terms used included: "Bone" AND "Marrow" AND "Aspirate" AND "Concentrate" AND "Bone Defects" AND "NONUNION". Eligible studies were independently appraised by two authors using the Critical Appraisal Skills Program checklist. For the purpose of narrative review, relevant studies were included irrespective of methodology or level of evidence. RESULTS: Thirty-four of the 103 (48 PubMed and 55 EMBASE) results yielded by the preliminary search were included. Exclusions included three duplicate records, six letters, 17 non-orthopaedics related studies and four records irrelevant to our search topic. The CASP appraisal confirmed a satisfactory standard of 31 studies. They all had clearly defined objectives, were well designed and conducted appropriately to meet them. The published studies reported the use of BMAC in non-union and fracture healing (15 studies), bone defects (nine studies), spine fusion (two studies), distraction osteogensis (two studies) and complications related to the use of BMAC (seven studies). CONCLUSIONS: Stem cells found in BMAC have the potential to self-renew, undertake clonal expansion and differentiate into different musculoskeletal tissues. The commercial processing of BMAC needs to be optimized in order to achieve a consistent end product, which will provide predicable and translatable results. The future potential of cell characterization in order to determine the optimum cell for repair/regeneration of bone also needs to be explored. LEVEL OF EVIDENCE: Systematic Review of minimum level IV studies.

3D-modeling of the spine using EOS imaging system: Inter-reader reproducibility and reliability.

OBJECTIVES: To retrospectively assess the interreader reproducibility and reliability of EOS 3D full spine reconstructions in patients with adolescent idiopathic scoliosis (AIS). METHODS: 73 patients with mean age of 17 years and a moderate AIS (median Cobb Angle 18.2°) obtained low-dose standing biplanar radiographs with EOS. Two independent readers performed "full spine" 3D reconstructions of the spine with the "full-spine" method adjusting the bone contour of every thoracic and lumbar vertebra (Th1-L5). Interreader reproducibility was assessed regarding rotation of every single vertebra in the coronal (i.e. frontal), sagittal (i.e. lateral), and axial plane, T1/T12 kyphosis, T4/T12 kyphosis, L1/L5 lordosis, L1/S1 lordosis and pelvic parameters. Radiation exposure, scan-time and 3D reconstruction time were recorded. RESULTS: Interclass correlation (ICC) ranged between 0.83 and 0.98 for frontal vertebral rotation, between 0.94 and 0.99 for lateral vertebral rotation and between 0.51 and 0.88 for axial vertebral rotation. ICC was 0.92 for T1/T12 kyphosis, 0.95 for T4/T12 kyphosis, 0.90 for L1/L5 lordosis, 0.85 for L1/S1 lordosis, 0.97 for pelvic incidence, 0.96 for sacral slope, 0.98 for sagittal pelvic tilt and 0.94 for lateral pelvic tilt. The mean time for reconstruction was 14.9 minutes (reader 1: 14.6 minutes, reader 2: 15.2 minutes, p<0.0001). The mean total absorbed dose was 593.4µGy ±212.3 per patient. CONCLUSION: EOS "full spine" 3D angle measurement of vertebral rotation proved to be reliable and was performed in an acceptable reconstruction time. Interreader reproducibility of axial rotation was limited to some degree in the upper and middle thoracic spine due the obtuse angulation of the pedicles and the processi spinosi in the frontal view somewhat complicating their delineation.

Core Decompression and Autologous Bone Marrow Concentrate for Treatment of Femoral Head Osteonecrosis: A Randomized Prospective Study.

The aim of this study was to investigate the safety of injection of bone marrow aspirate concentrate during core decompression and to study its clinical (visual analogue scale; Harris-Hip-score) and radiological outcomes (magnetic resonance imaging). In this prospective and randomized clinical trial we evaluated 24 consecutive patients with non-traumatic femoral head necrosis (FHN) during a period of two years after intervention. In vitro analysis of mesenchymal stem cells was performed by evaluating the fibroblast colony forming units (CFU-Fs). Postoperatively, significant decrease in pain associated with a functional benefit lasting was observed. However, there was no difference in the clinical outcome between the two study groups. Over the period of two years there was no significant difference between the head survival rate between both groups. In contrast to that, we could not perceive any significant change in the volume of FHN in both treatment groups related to the longitudinal course after treating. The number of CFU showed a significant increase after centrifugation. This trial could not detect a benefit from the additional injection of bone marrow concentrate with regard to bone regeneration and clinical outcome in the short term.

Primary stability of the Fitmore stem: biomechanical comparison.

PURPOSE: After clinical introduction of the Fitmore stem (Zimmer), we noticed the formation of cortical hypertrophies in a few cases. We questioned whether (1) the primary stability or (2) load transfer of the Fitmore stem differs from other stems unassociated with the formation of hypertrophies. We compared the Fitmore stem to the well-established CLS stem. METHODS: Four Fitmore and four CLS stems were implanted in eight synthetic femurs. A cyclic torque around the stem axis and a mediolateral cyclic torque were applied. Micromotions between stems and femurs were measured to classify the specific rotational implant stability and to analyse the bending behaviour of the stem. RESULTS: No statistical differences were found between the two stem designs with respect to their rotational stability (p = 0.82). For both stems, a proximal fixation was found. However, for the mediolateral bending behavior, we observed a significantly (p < 0.01) higher flexibility of the CLS stem compared to the Fitmore stem. CONCLUSION: Hip stem implantation may induce remodelling of the periprosthetic bone structure. Considering the proximal fixation of both stems, rotational stability of the Fitmore® stem might not be a plausible explanation for clinically observed formation of hypertrophies. However, bending results support our hypothesis that the CLS stem presumably closely follows the bending of the bone, whereas the shorter Fitmore stem acts more rigidly. Stem rigidity and flexibility needs to be considered, as they may influence the load transfer at the implant-bone interface and thus possibly affect bone remodelling processes.

Regulation of pro-angiogenic tissue factor expression in hypoxia-induced human lung cancer cells.

Alternative splicing is a key regulatory mechanism for cellular metabolism controlling cell proliferation and angiogenesis, both of which are crucial processes for tumorigenesis under hypoxia. Human cells express two tissue factor (TF) isoforms, alternatively spliced TF (asTF) and 'full length' TF (flTF). flTF is the major source of thrombogenicity whereas, the function of soluble asTF, particularly in cancer, is widely unknown. In the present study, we examined the impact of alternative splicing on the pro-angiogenic potential and the TF expression pattern of A549 cells under hypoxia. We focused our efforts toward alternative splicing factors, such as Clk1, and pro-angiogenic proliferation-regulating factors, such as Cyr61. We further examined the influence of asTF overexpression on the expression of MCP-1, Cyr61 and VEGF, as well as on cell number and pro-angiogenic properties of A549 cells. Notably, we found hypoxia to induce the expression of alternative splicing factors (Clk1 and Clk4) as well as proliferation- and angiogenesis-promoting factors (Cyr61 and flTF). asTF overexpression in A549 cells also increased both cell number and tube formation. These effects were mediated by the induction of Cyr61, MCP-1 and VEGF, as well as by integrin α(v)ß(3). Taken together, our results suggest that the pro-angiogenic potential of A549 lung cancer cells is modulated under hypoxic conditions via modulation of TF isoform expression which in turn is controlled by alternative splicing.

Haematogenous infection of a total knee arthroplasty with Klebsiella pneumoniae.

This case report describes a prosthetic joint infection due to the haematogenous spread of Klebsiella pneumoniae from a genitourinary focus. Prior to the infection, the patient was diagnosed with early stage prostatic carcinoma, which had been successfully treated with surgery. However, in the time period following surgical treatment, the patient suffered recurring urinary tract infections. During the course of these recurring infections, he developed a concurrent bacterial infection of his total knee arthroplasty. Two sequential joint aspirates revealed K pneumoniae to be the cause. Therefore, two-stage revision total knee arthroplasty was performed. This case reiterates the fact that invasive therapeutic procedures can cause bacteraemia resulting in infection of a joint replacement. We would therefore like to emphasise the importance of prophylactic antibiotic treatment prior to invasive therapies, particularly in organs with potentially large counts of bacteria.

Bivalirudin inhibits periprocedural platelet function and tissue factor expression of human smooth muscle cells.

AIM: A major concern of stent implantation after percutaneous coronary intervention (PCI) is acute stent thrombosis. Effective inhibition of periprocedural platelet function in patients with coronary artery disease (CAD) leads to an improved outcome. In this study, we examined the periprocedural platelet reactivity after administrating bivalirudin during PCI compared to unfractionated heparin (UFH) administration. Further, the effect of bivalirudin on induced tissue factor (TF) expression in smooth muscle cells (SMC) was determined. METHODS: Patients with CAD (n = 58) and double antithrombotic medication were treated intraprocedural with UFH (n = 30) or bivalirudin (n = 28). Platelet activation markers were flow cytometrically measured before and after stenting. The expression of TF in SMC was determined by real-time PCR and Western blotting. The thrombogenicity of platelet-derived microparticles and SMC was assessed via a TF activity assay. RESULTS: Bivalirudin significantly diminished the agonist-induced platelet reactivity post-PCI. Compared to UFH treatment, the adenosine diphosphate (ADP) and thrombin receptor-activating peptide (TRAP)-induced thrombospondin expression post-PCI was reduced when bivalirudin was administrated during intervention. In contrast to UFH, bivalirudin reduced the P-selectin expression of unstimulated and ADP-induced platelets post-PCI. Moreover, bivalirudin inhibited the thrombin-, but not FVIIa- or FVIIa/FX-induced TF expression and pro-coagulant TF activity of SMC. Moreover, bivalirudin reduced the TF activity of platelet-derived microparticles postinduction with TRAP or ADP. CONCLUSIONS: Bivalirudin is better than UFH in reducing periprocedural platelet activation. Moreover, thrombin-induced TF expression is inhibited by bivalirudin. Thus, bivalirudin seems to be a better anticoagulant during PCI than UFH.

Prolonged clopidogrel application reduces tissue factor expression after percutaneous coronary intervention in the porcine model.

BACKGROUND: Late thrombotic events are important complications associated with intracoronary brachytherapy (ICBT) using ionizing radiation (IR) or with antiproliferative treatment modalities such as drug-eluting stents (DES). The mechanism mediating these thrombotic events is not well understood. This study assessed the effect of prolonged clopidogrel treatment on tissue factor (TF) expression in coronary arteries and on the circulating TF level after percutaneous transluminal coronary angioplasty /ICBT in a porcine coronary model. METHODS: Pigs were treated with aspirin plus a 300 mg loading dose of clopidogrel one day before percutaneous coronary intervention (PCI), followed by a daily dose of clopidogrel and aspirin. During PCI one of the two balloon-injured arteries was treated by brachytherapy. Animals were sacrificed at different time points. The pigs, which were sacrificed 3 months post-PCI, were divided into two groups (Group I: clopidogrel for 3 months; Group II: clopidogrel for 1 month). Plasma TF was measured by enzyme-linked immunosorbent assay in blood samples taken from all pigs before and immediately after intervention and before sacrifice. Morphometric analysis was performed on digitalized images employing the software LUCIA G for TF staining. Vascular TF expression levels were assessed by quantitative real-time polymerase chain reaction. RESULTS: Prolonged clopidogrel application significantly reduced coronary TF at the protein (Group I vs. II, 8.975 ± 3.947% vs. 26.44 ± 5.375%, P = .007) and mRNA level [Group I vs. II, (0.3501 ± 0.0519) × 10(-3) vs. (0.7073 ± 0.0436) × 10(-3), P<.0005]. Circulating TF protein tended to be lower after 3 months than after 1 month clopidogrel treatment post-PCI (Group I vs. Group II, 488.3 ± 35.37 pg/ml vs. 572.3 ± 39.9 pg/ml, P = .130). CONCLUSIONS: Prolonged clopidogrel treatment reduced coronary TF expression and tended to reduce the blood TF level post-PCI, thus possibly modulating the risk of late thrombosis.

Role of the phosphatidylinositol 3-kinase/protein kinase B pathway in regulating alternative splicing of tissue factor mRNA in human endothelial cells.

BACKGROUND: Tissue factor (TF) is the primary initiator of blood coagulation. In response to tumor necrosis factor (TNF)-alpha human umbilical vein endothelial cells (HUVECs) express 2 TF isoforms: a soluble alternatively spliced isoform (asHTF) and membrane-bound "full length" (fl)TF. How the differential TF isoform expression is regulated is still unknown. This study compared the impact of PI3K/Akt pathway inhibition on alternative splicing of TF in HUVECs, to the influence of transcriptional regulation by inhibiting nuclear factor kappa B (NFkappaB). METHODS AND RESULTS: The mRNA expression of TF isoforms was assessed by real-time PCR, the thrombogenic activity was measured by a chromogenic TF activity assay and the phosphorylation state of serine/arginine-rich (SR) proteins was analyzed by western blotting. Transfection of HUVECs was done 72 h before the inhibition experiments were performed. PI3K/Akt pathway inhibition reduced the mRNA expression of asHTF but not flTF. Inhibition of NFkappaB reduced the expression of both isoforms. Moreover, the PI3K/Akt pathway inhibition, but not that of NFkappaB, modified the phosphorylation of the SR proteins SRp75, SRp55 and SF2/ASF. Additionally, overexpression of SF2/ASF and SRp75 influenced the differential TF-isoform expression in HUVECs. CONCLUSIONS: The PI3K/Akt pathway modulates alternative splicing of TF in HUVECs, distinct from transcriptional regulation.