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Biomedical subjects

M van Griensven

Publications and source records attributed to M van Griensven.

At least 19 recordsLinked to original sources

Influence of cyclic mechanical strain and heat of human tendon fibroblasts on HSP-72.

Heat shock protein 72 (HSP-72) is a member of a superfamily of different proteins that are synthesized as a cytoprotective response following cellular stress. Mechanical strain is an important component in ligament and tendon healing. Up to the present point of time, the influence of mechanical strain on the expression of HSP-72 is unknown. Tendon fibroblasts from the patellar tendons of nine individuals were isolated and amplified in vitro. First, the effect of 15 or 60 min of heat exposition was studied immunohistochemically and by Western blotting. In a second experiment, the effects of 15 and 60 min of cyclic longitudinal stretching were investigated. Samples were taken after 2, 4 and 8 h. The heat exposition experiments indicate that HSP-72 accumulates in the nucleus and that there is a transient upregulation. This effect is more prominent after 60 min of heat exposure. The same reaction was found after stretching stimulation, however, to a lesser extent. There was a transient up regulation of HSP-72 after short-term stretching and a biphasic increase after 60 min of stretching. Upregulation of HSP-72 by heat and mechanical stress is a response in human fibroblasts which involves a nuclear translocation. The response differs with regard to the time points beyond 2 h after the application of either stress.

Adolescent↗

A comparison of two types of free bone grafts as transport discs in segmental distraction for reconstruction of calvarial bone defects: an experimental study.

INTRODUCTION: Several investigations over the past few years have shown that the importance of preserving the blood supply of the transport disc during segmental distraction has been overestimated. It was clearly demonstrated that distraction osteogenesis by local bone transportation could be achieved even with free bone grafts. So far, there have been no systematic investigations into how different types of free bone grafts vary in their function and suitability as transport discs in segmental craniofacial distraction. The aim of this study was to develop criteria to aid the clinician in selecting the most suitable free bone graft for distraction purposes. MATERIALS AND METHODS: Under general anesthesia, calvarial defects measuring 6 x 5 cm were created in 12 adult black head sheep in order to reconstruct them by segmental distraction using two different types of bone grafts as transport discs. Group 1 (n=6) received autotopical membranous bone of the calvarium as the transport segment, while group 2 (n=6) received heterotopical enchondral material from the iliac crest. Distraction was started postoperatively after 5 days at 0.6 mm/day and continued for 7 weeks. After a further 6 weeks of consolidation, the animals were killed and specimens examined macroscopically, radiographically and histologically. RESULTS: The significantly better quality of bone regenerates of group 1 was clearly influenced by the significantly better stability of the transport disc's connection to the distraction device, the calvaria grafts showing better volume stability and better mechanical resistance during transport than the iliac bone. The influence of both types of bone grafts on the macro- and microstructure of the newly formed bone was clearly evident. The mineralization density of group 2 regenerates was significantly lower than that of group 1 specimens. CONCLUSION: High mechanical stability of the bone graft is a very important point to consider when selecting a free graft for use as transport disc in local bone transport, especially if transport is necessary over long distances.

Animals↗

Effects of cyclic longitudinal mechanical strain and dexamethasone on osteogenic differentiation of human bone marrow stromal cells.

The aim of the study was to investigate the effect of cyclic mechanical strain on differentiation markers in the presence or absence of dexamethasone. Human bone marrow stromal cells (BMSC) from seven donors (32.5+/-6.2 years) were cultivated with (D+) or without (D-) dexamethasone. A cyclic mechanical strain with an elongation of 2% (D+2; D-2) or 8% (D+8; D-8) was applied for three days with a stimulation time of three times two hours each day. Levels of alkaline phosphatase (ALP) and osteocalcin (OC) were compared after time intervals of four and seven days. mRNA expression of Collagen I, III and Cbfa1 was investigated after one, four, and seven days. ALP levels were significantly increased in the D+8 group after four and seven days (147.1+/-6.3%; p<0.05 and 168.6+/-6,5%; p<0.03) and in the D-8 group after 7 days (197.4+/-10.4; p<0.04). Cyclic strain had a significant influence on ALP-secretion (F=7.5; p<0.01). In the D-8 group there was a significant increase in OC secretion after 4 days (140.9+/-12.5%; p<0.05).; p<0.01). The effect of stretching was significantly stronger than that of dexamethasone (F=17.2 vs. 1.8). Collagen I (Col I) expression was upregulated in D+8 cultures after 4 days (215.0+/-53.3 p<0.04) and after seven days (166.7+/-55.7; p<0.04). Collagen III (Col III) expression was upregulated in D+2 and D+8 cultures after 4 days (200.7+/-16.3 and 185.9+/-12.7; p<0.04) and after seven days (154.4+/-10.1 and 118.8+/-16.4; p<0.04). There was a significant increase of Cbfa1 expression in D+8 cultures at all investigated time intervals (day 1: 105.5+/-3.7%; day 4: 104.7+/-3.0%; day 7: 104.4+/-2.1%; p<0.03). Stretching (F=20.0; p<0.01) was a stronger contributor to Cbfa-1 expression than dexamethasone (F=12.1; p<0.01). Cyclical mechanical stimulation with 8% elongation increases ALP and OC levels and upregulates Col I and III synthesis and Cbfa1 expression. In the short term, cyclical stretching is a stronger differentiation factor than dexamethasone. Cyclical stretching and dexamethasone both enhance the osteogenic commitment of hBMSC.

Adult↗

Scaphotrapeziotrapezoid (STT)-arthrodesis in Kienbock's disease.

This study reviews the results of 59 of 84 patients with severe Kienbock's disease who were treated with STT fusion. The average follow-up period was 4 (ranges: 2-8 years). The average arc of wrist extension and flexion was 67 degrees (60% of the contralateral side, 81% of pre-operative range) and that of ulnar and radial deviation was 31 degrees (52% of the contralateral side, 56% of pre-operative range). Pre-operative pain values (VAS) were 56 (non-stress) and 87 (stress) and were significantly higher than the postoperative values of 12 (non-stress) and 41 (stress). Grip strength improved from 45kPa pre-operatively to 52kPa postoperatively. The mean modified Mayo wrist score was 63 points. The patients reported low disability in the DASH scores, with an average of 28 points. Our data show that STT fusion is a reliable and effective treatment for pain relief and offers a good functional result in advanced stages of Kienbock's disease. However the long-term effect of this procedure on radioscaphoid and other intercarpal joints is yet to be determined.

Adolescent↗

Are alterations of lymphocyte subpopulations in polymicrobial sepsis and DHEA treatment mediated by the tumour necrosis factor (TNF)-alpha receptor (TNF-RI)? A study in TNF-RI (TNF-RI(-/-)) knock-out rodents.

Sepsis is associated with depression of T cell-dependent immune reactivity with proinflammatory cytokines, such as tumour necrosis factor (TNF)-alpha, playing an important role. Recent investigations describe an association between these immunological alterations and disturbances of the endocrine system, related most frequently to sex steroid hormones. Dehydroepiandrosterone (DHEA), one of the most abundant adrenal sex steroid precursors, seems to have a protective immunological effect towards septic insults. In this study, both the role of TNF-receptor I (RI) and possible interactions in the protective role of DHEA were investigated in a murine model of polymicrobial sepsis. Polymicrobial sepsis was induced by caecal ligation and puncture (CLP) in a murine model. The effects of DHEA on survival, clinical parameters and cellular immunity (T lymphocytes and natural killer (NK) cells) were investigated. CLP was performed in genetically modified TNF-RI knock-out (TNF-RI(-/-)) and genetically unmodified (wild-type, WT) mice. DHEA application was associated with a decrease in the mortality rate in WT animals. A mortality rate of 91.7% was observed in TNF-RI(-/-) mice after CLP. This mortality rate was reduced to 37.5% by the application of DHEA. In sham-operated TNF-RI(-/-) animals, a significantly higher proportion of NK cells within the lymphocyte population was measured compared with the corresponding WT group. After CLP, a significant increase in the percentage cell count of NK cells was recorded in WT mice. Overall, following DHEA application in WT mice, an alteration in the cellular immune response was characterized by a reduction in the percentage counts of CD4(+), CD8(+) and NK cells. In the group of TNF-RI(-/-) mice treated with DHEA, no increase in the percentage cell count of NK cells was observed after CLP. No data for cell analysis were available from the CLP-TNF-RI(-/-) mice treated with saline, due to the high mortality rate in these animals. DHEA reduces the complications of sepsis in a TNF-RI-independent manner. Our study suggests that NK cells are involved in the protective mechanism of DHEA in WT mice. It would therefore seem that DHEA represents a feasible alternative therapy for the dysregulated immune system in sepsis.

Animals↗

Comparison of human bone marrow stromal cells seeded on calcium-deficient hydroxyapatite, beta-tricalcium phosphate and demineralized bone matrix.

The aim of this study was to compare three resorbable biomaterials regarding seeding efficacy with human bone marrow stromal cells (BMSCs), cell penetration into the matrix, cell proliferation and osteogenic differentiation. Calcium-deficient hydroxyapatite (CDHA), beta-tricalcium phosphate (beta-TCP), and demineralized bone matrix (DBM) were seeded with human BMSCs and kept in human serum and osteogenic supplements for 3 weeks. Morphologic and biochemical evaluations were performed on day 1, 7, 14 and 21. The allograft DBM and CDHA exhibited both an excellent seeding efficacy while the performance of beta-TCP was lower when compared. The total protein content and the values for specific alkaline phosphatase (ALP) increased on all matrices and no significant difference was found for these two markers. BMSCs in monolayer had a significant increase of protein, but not of ALP. Osteocalcin (OC) values increased significantly higher for BMSC in cultures on DBM when compared to CDHA and beta-TCP. The OC levels decreased significantly in the BMSC monolayer culture. BMSCs were found inconsistently within the synthetic materials, whereas in DBM they were found more homogeneously distributed throughout the matrix. All three matrices promoted BMSC proliferation and differentiation to osteogenic cells. DBM allografts seem to be more favorable with respect to cell ingrowth tested by histology, and osteogenic differentiation ascertained by an increase of OC. CDHA with its high specific surface area showed more favorable properties than beta-TCP regarding reproducibility of the seeding efficacy.

Biocompatible Materials↗

[Effect of cyclical stretch on matrix synthesis of human patellar tendon cells].

The significance of mobilization and loading for healing ligaments and tendons is generally accepted today. Local deformation of cells thereby represents the key stimulus for the cellular response. Less is known, however, about the effects of cyclic strain on the cellular and molecular level. The aim of the in vitro investigation was to determine the effect of cyclic mechanical strain on collagen type I and III and on fibronectin formation in human patellar tendon derived fibroblasts. Human patellar tendon derived fibroblasts from 5 donors (mean age 29.2 years) were cultured under standard conditions. Monolayers of subconfluently grown 3rd passage cells were stretched in rectangular silicone dishes with cyclic movement along their longitudinal axes. Cyclic strain (5%, 1 Hz) was applied for 30 min and 60 min, respectively. Carboxyterminal procollagen type I propeptide (P-I-CP) and aminoterminal procollagen type III propeptide (P-III-NP) release was measured by a radio-immunoassay 6 h and 12 h after stretching. The release of fibronectin was measured by nephelometry following immunoreaction with a specific antiserum. Cells from each donor without any cyclic stretching served as controls. Compared with the controls, only cyclic stretching for 60 min resulted in a significantly increased release of P-I-CP and fibronectin after 6 h. The release of P-III-NP was significantly increased 12 h after 30 min of cyclic stretching as well as 6 h and 12 h after 60 min of cyclic stretching, respectively. We conclude that cyclic stretching causes a time-dependent, differential regulation of formation of fibronectin and collagen type I and III. This may effect the quality and thus the mechanical properties of healing tendon and ligament tissues. In order to improve current treatment protocols and to enlarge our knowledge of tissue healing, it is necessary to understand the cellular response to cyclic strain.

Adult↗

Pulmonary microbial infection in mice: comparison of different application methods and correlation of bacterial numbers and histopathology.

Many investigations have been performed in characterising experimental bacterial infections in the lung of mice using several pathogens. Robust experimental pulmonary infection models require a reproducible method of application with defined numbers of pathogens to the respiratory tract without contaminating extrapulmonary tissues. At the same time trauma due to the experimental procedure should be kept to a minimum. So far several routes of administration have been used but a systematic comparison of these methods is still missing. Here we provide a comprehensive evaluation of view controlled i.t. instillation, tracheotomy, intranasal application, blind instillation and aerosol infection. An infection dose of up to 5 x 10(4) bacteria (L. monocytogenes) was applied to a group of ten mice by each technique and the animals were killed after 1 h or 24h. The number of viable bacteria was estimated by plating homogenates of the lungs and spleens. In addition, pathological effects on lung tissue were examined by histology 24h after infection. The highest reproducibility was achieved after applying Listeria directly in the trachea under view or by tracheotomy. However, mice were severely affected in their vitality after tracheotomy. Thus, for topical application of bacterial suspension into the lung the view controlled i.t. instillation is most appropriate.

Animals↗

Adenoviral gene transfer in a rat fracture model.

For the enhancement of fracture healing, either purified proteins or vectors for expression of growth factors in situ may be used. Adenoviral vectors directly convert cells to express a transgene. However, the cell types which are preferentially infected and the time of expression during fracture healing are currently not known. The adenoviral type 5 vectors used in this study are replication incompetent viruses, one encoding beta-galactosidase (beta-GAL) and one green fluorescent protein. Femora of 35 Sprague-Dawley rats were fractured. Three days after stabilization with Kirschner wire, 10(12) pfu viral suspension were injected into the fracture zone. As a control, five animals received injections of adenovirus type 2. Animals were sacrificed after 3 days, 1, 2 and 4 weeks. Fractures healed radiographically within 2-3 weeks. All specimens were examined for beta-GAL and green fluorescent protein (GFP) expression. Fibroblast and osteoblasts within callus tissue displayed a high transgene expression (week 1). A decrease of expression was observed during the observation period. In this experimental study, we have demonstrated that all cells of the primary callus can be transfected using adenoviral vectors, which provide a tool to further investigate adenoviral transfer of growth factors such as bone morphogenetic protein-2 (BMP-2).

Adenoviridae↗

[Pronation and supination of the forearm. With special reference to the humero-ulnar articulation].

The development of a kinematic model of the pro- and supination, that can be used to predict the influence of angulations of ulna and radius on the pronation and supination is based on the precise knowledge of the pronation and supination movement. We performed two parallel studies for examining the pronation and supination motion of the human forearm. The first experiment dealt with MRI-studies on 18 probands (36 examined forearms). As a result we observed an evasive movement of the ulna during the rotation of 7, 14 degrees medial. In order to prove whether the evasive movement was caused by a rotation of the humerus or by an evasion in the articulatio humeroulnaris, we carried out a second experiment, using 30 preparations. The measurement of the pro- and supination motion with a fixed humerus was expedited using a special experimental setup which guaranteed that the ulna could move freely. In all cases we found the same magnitude of the evasive motion of the ulna. Therefore we demonstrated, that the ulna performs an evasive motion during the pro- and supination motion of the forearm that influences the kinematic behavior of the pro- and supination motion significantly.

Adult↗

Cyclic mechanical stretching enhances secretion of Interleukin 6 in human tendon fibroblasts.

Accelerated rehabilitation after tendon and ligament injuries is widely accepted to avoid adverse effects of immobilization. However, progressive rehabilitation may also lead to an excessive inflammatory soft tissue response. To investigate the amount of loading necessary to accelerate the healing process without causing damage to the healing tissue, we experimentally stretched human tendon fibroblasts of healthy tendons 15 and 60 min with 1 Hz and an elongation of 5% and measured the secretion of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta1 (TGF-beta1), platelet-derived growth factor (PDGF), and fibroblast growth factor basic (bFGF). Secretion of IL-6 was significantly induced by 15 min of cyclic biaxial mechanical stretching after 4 and 8 h observation time and by 60 min stretching and 2 h observation time. The growth factors TGF-beta1, bFGF, and PDGF were secreted by human tendon fibroblasts both in stretched cells and controls; however, no increases were related to mechanical stretching. There was no measurable secretion of TNF-alpha in human tendon fibroblasts. These findings suggest that the inflammatory reaction often seen during physiotherapy after tendon and ligament injuries is caused in part by secretion of IL-6 from the stretched human tendon fibroblasts. IL-6 may cause exaggerated proliferation of fibroblasts and synovial cells as seen in rheumatoid arthritis and arthrofibrosis. However, physiological proliferative reactions leading to repair of injured tissue are also possible. IL-6 measured in the synovial fluid may be an important predictor for monitoring and improving therapeutic strategies in terms of tendon/ligament healing.

Cell Line↗

Arthrofibrosis is the result of a T cell mediated immune response.

It is thought that an excessive fibrotic healing response with diffuse intra-articular scarring leads to arthrofibrosis after trauma and surgery around joints. To clarify the specific cellular mechanism of arthrofibrosis during arthrolysis we took fibrotic tissue samples from 18 patients at varying periods after knee trauma or surgery. Sections were stained with hematoxylin and eosin to study the overall histopathological changes. Major histocompatibility complex (MHC) class II expressing cells as well as CD3, CD4, CD25, CD28, CD68, CD80, and CD83 positive cells were localized immunohistologically. The results demonstrated synovial hyperplasia with fibrotic enlargement of the subintima and infiltration of inflammatory cells. The number of MHC class II expressing cells was increased. Mainly, intimal macrophages and dendritic cells showed positive immunostaining for MHC class II antigens. In the subintima moderate infiltration of T cells including activated T cells (CD25), CD4+ T helper (Th) cells and Th1 and Th2 subsets was detected. There was a slight polarization of the Th1/Th2 balance towards Th1 differentiation. Positive immunostaining for CD80/CD28 indicated the costimulatory signal for T cell activation and clonal expansion. These findings strongly support an immune response as the cause of capsulitis leading to formation of diffuse scar tissue within the knee joint. Based on our immunohistological study we conclude that a T cell mediated immune response plays a crucial role in the mechanism of arthrofibrosis.

Adolescent↗

Cyclic mechanical stretching modulates secretion pattern of growth factors in human tendon fibroblasts.

The objective of the study was to investigate whether the response profile of the growth factor of human tendon fibroblasts could be beneficially influenced through the application of mechanical stretch. It was considered that this would elucidate structural and functional problems, often seen after tendon and ligament healing. The secretion pattern of transforming growth factor-beta (TGF-beta), platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF) was determined in mechanically stretched fibroblasts and compared to non-stretched controls. Human tendon fibroblasts were experimentally stretched for 15 and 60 mm at a frequency of 1 Hz and an amplitude of 5%. The secretion of TGF-beta PDGF and bFGF was measured by enzyme-linked immuno-sorbent assay. All the growth factors investigated were indeed secreted by human tendon fibroblasts both in stretched cells and controls. Mechanical stretch increased the secretion pattern of the growth factors. The increased concentrations of TGF-beta bFGF and PDGF after cyclical mechanical stretching may have a positive influence on tendon and ligament healing through stimulation of cell proliferation, differentiation and matrix formation.

Adult↗

Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis.

OBJECTIVE: Sepsis is associated with a marked depression of cellular immune function. The steroid hormone dehydroepiandrosterone (DHEA) is proposed to have immunoenhancing activities. We, therefore, investigated the effect of DHEA on the mortality rate and cellular immune functions in an experimental model of sepsis. DESIGN: Randomized animal study. SETTING: Level I trauma center, university research laboratory. SUBJECTS: Male NMRI mice. INTERVENTIONS: Mice were subjected to laparotomy (sham) or cecal ligation and puncture (CLP). Mice were treated with (sham/DHEA; CLP/DHEA) or without (sham; CLP) the steroid hormone DHEA (30 mg/kg sc). Animals were killed 48 hrs after the onset of sepsis. MEASUREMENTS AND MAIN RESULTS: The survival rate of septic mice was determined 24 and 48 hrs after onset of sepsis. Forty-eight hours after the septic challenge, a white blood cell count was performed and serum tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta concentrations were monitored using ELISA. Furthermore, the delayed type of hypersensitivity (DTH) reaction was evaluated on the basis of ear pinna swelling after dinitrofluorobenzene (DNFB) administration, and clinical variables (body weight, temperature, heart rate, fluid input/output, food intake) were monitored using metabolic cages. DHEA administration improved the survival rate (87% vs. 53% after 48 hrs; p <.001). This was accompanied by a restoration of the depressed DTH reaction and a reduction in TNF-alpha serum concentrations (20.7 +/- 1.4 pg/mL vs. 32.4 +/- 6.6 pg/mL). CONCLUSIONS: These results demonstrate that DHEA administration leads to an increased survival following a septic challenge. The immunoenhancing effect of DHEA is accompanied by a reduction of TNF-alpha release and an improved activity of T-cellular immunity. DHEA administration may, therefore, be beneficial in systemic inflammation.

Adjuvants, Immunologic↗

Heterotopic ossifications in patients after severe blunt trauma with and without head trauma: incidence and patterns of distribution.

OBJECTIVE: To investigate the incidence and distribution of heterotopic ossifications in patients with blunt multiple trauma with and without associated head trauma. DESIGN: Retrospective. SETTING: Level I trauma center. PATIENTS: Patients were included if they were treated between August 1987 and September 1995. Inclusion criteria included age between 16 and 65 years, injury severity score (ISS) of more than twenty points, and clinical reexamination performed more than three years after the initial injury. METHODS: The records of each patient were abstracted to determine the ISS, the Glasgow coma score (GCS), and parameters describing the course of intensive care. For each patient, a reexamination was performed between January and September 1998. Patients with multiple trauma and associated head trauma (Group PTH, polytrauma, GCS less than nine points, and head computed tomography scan abnormalities) and patients with multiple trauma without associated head trauma (Group PT, polytrauma, GCS of at least nine points, and normal head computed tomography scans) were compared. A clinical reexamination was performed to evaluate functional outcome. RESULTS: Sixty-four patients belonged to Group PTH and 124 patients belonged to Group PT. There were no differences in the age (Group PTH, 28.9 +/- 1.6 years; Group PT, 29.2 +/- 2.1 years) or severity of injury (ISS Group PTH, 31.0 +/- 5.3 points; ISS Group PT, 33.0 +/- 6.1 points) among patients in the two groups. The overall incidence of periarticular heterotopic ossification was comparable in patients with multiple trauma with and without head injury (Group PTH, 30 of 64 patients [46.9 percent]; Group PT, 53 of 124 patients [42.7 percent]). The duration of ventilation was significantly higher in Group PT (Group PTH, 9.3 +/- 2.4 days; Group PT, 14.2 +/- 3.1 days; p = 0.02). In the subgroups in which heterotopic ossification developed (PT-HO and PTH-HO), patients in PT-HO had a significantly higher incidence of heterotopic ossification, as compared with patients in PTH-HO at initially uninjured joints (Group PTH-HO, 1 of 30 patients [3.3 percent]; Group PT-HO, 10 of 53 patients [18.9 percent]; p = 0.04). CONCLUSIONS: There was a high incidence of heterotopic ossification around those joints that were initially classified as uninjured in patients without head trauma. This finding suggests that pathogenic pathways independent of head trauma, such as long-term ventilation, play a main role. Causative factors for the development of heterotopic ossification at initially uninjured joints in long-term ventilated patients with multiple trauma with and without head trauma remain to be elucidated.

Adult↗

Major secondary surgery in blunt trauma patients and perioperative cytokine liberation: determination of the clinical relevance of biochemical markers.

BACKGROUND: The aim of this study is to assess the associations between the timing of secondary definitive fracture surgery on inflammatory changes and outcome in the patient with multiple injuries. The study population consists of a series of patients with multiple injuries who were managed using a strategy of primary temporary skeletal stabilization followed by delayed definitive fracture fixation. METHODS: In a prospective cohort study performed at a Level I trauma center, the patients' injuries and operative details as well as immune markers and clinical outcomes were studied. The patients were split into an early secondary surgery group (group ESS, surgery at days 2-4) and a late secondary surgery group (group LSS, surgery at days 5-8). During the posttraumatic course, inflammatory markers (interleukin [IL]-6, tumor necrosis factor-alpha) were determined on a daily basis. Perioperatively, these markers were additionally evaluated at 30 minutes, 7 hours, and 24 hours after initiation of surgery. RESULTS: Secondary surgery on days 2 to 4 was associated with a higher incidence of postoperative organ dysfunction (n = 33 [46.5%]) than secondary surgery on days 5 to 8 (n = 9 [15.7%], p = 0.01). A significant association between the combination of initial IL-6 values > 500 pg/dL plus surgery on days 2 to 4 and the development of multiple organ failure (r = 0.96, p < 0.001) occurred. A correlation between the initial IL-6 values > 500 pg/dL and surgery on days 5 to 8 (r = 0.57, p < 0.07) could not be found. IL-6 also demonstrated a predictive value for the development of multiple organ failure: IL-6 > 500 pg/dL in group ESS, r = 0.96, p < 0.001; IL-6 > 500 pg/dL in group LSS, r = 0.57, p < 0.07. CONCLUSION: According to our data, no distinct clinical advantage in carrying out secondary definitive fracture fixation early could be determined. In contrast, in patients who demonstrated initial IL-6 values above 500 pg/dL, it may be advantageous to delay the interval between primary temporary fracture stabilization and secondary definitive fracture fixation for more than 4 days. In patients with blunt multiple injuries undergoing primary temporary fixation of major fractures, the timing of secondary definitive surgery should be carefully selected, because it may act as a second hit phenomenon and cause a deterioration of the clinical status.

Adolescent↗

The effect of trauma on neutrophil L-selectin expression and sL-selectin serum levels.

Among identified adhesion molecules, the L-selectin on neutrophils enables the first step of leukocyte adherence to activated endothelial cells. To allow firm adhesion of neutrophils, L-selectin is then split off the cell membrane. It was hypothetized that an increase of the constitutively high serum level of soluble L-selectin may indicate an ongoing pathological neutrophil sequestration to the endothelial cells associated with activation and injury of the cells. To evaluate this hypothesis, sL-selectin serum levels and neutrophil L-selectin expression of healthy volunteers (group A, n = 15), as well as of surgical patients, were investigated. Group B (n = 26) included patients subjected to elective limb surgery (mean operation time, 122 min), and group C (n = 45) comprised trauma patients. sL-selectin serum levels were measured daily over a 14-day period. Neutrophil L-selectin expression was evaluated by FACS analysis using the humanized anti-L-selectin antibody HuDreg 55 over a period of 3 days at minimum in both experimental groups. The binding of sL-selectin to endothelial cells was also examined in vitro. Elective limb surgery resulted in lower pre- and post-operative sL-selectin plasma levels (800-1,000 ng/mL) compared to healthy volunteers (1,100-1,200 ng/mL) with insignificant changes throughout the study period. Trauma patients revealed even lower sL-selectin levels (400-600 ng/mL). When these patients were discriminated by the multiple organ dysfunction (MOD) score of Moore in +MOD (n = 9, ISS = 31.7) and -MOD (n = 36, ISS = 25.0), a significant difference became evident. In +MOD patients sL-selectin levels remained on a low basis of 350 ng/mL, whereas in -MOD patients the initial low sL-selectin level subsequently rose to 800 ng/mL, similar to that of elective surgery patients. FACS analysis revealed a significant drop in neutrophil L-selectin expression 24 h after trauma compared to normal. Also, +MOD and -MOD patients were significantly discriminated by the L-selectin expression at this time. The in vitro studies revealed evidence for binding of sL-selectin to endothelial cells independently on the presence of neutrophils. According to our data, increasing severity of the post-operative/posttraumatic course is associated with decreasing sL-selectin serum levels and also reduced neutrophil L-selectin expression. In view of the in vitro results, this probably indicates competitive enhanced binding of sL-selectin to endothelial cells, thus masking the elevated activation of neutrophils and their ability for endothelial adherence.

Adolescent↗