Surface proteins and osteoblast markers: characterization of human adipose tissue-derived osteogenic cells.

INTRODUCTION: In adipose tissue-derived osteogenic cells (ADOC), osteoblast markers and surface proteins were determined and compared with osteoblasts harvested from cancellous bone (OB). METHOD: Osteocalcin (OC), core binding factor 1 (CBFA1), collagen type 1 (Coll1), alkaline phosphatase (ALP), nucleostemin (NS), and surface proteins CD 10, CD44, CD 59 and CD 105 were analyzed using RT-PCR, immunofluorescence and Western blot. RESULTS: Osteocalcin expression was more distinct in OB than in ADOC, but the other markers and surface proteins showed no differences. CONCLUSION: These data support the use of adipose tissue for future regenerative medicine; however, further studies are necessary to establish the role of long-term differentiation.

Effect of low molecular weight heparin (dalteparin) and fondaparinux (Arixtra) on human osteoblasts in vitro.

BACKGROUND: The prolonged administration of heparin for prevention and treatment of venous thromboembolism has been associated with a risk of heparin-induced osteoporosis. Fondaparinux is a new antithrombotic drug that specifically inhibits factor Xa. Because of the known interactions of other antithrombotic agents with bone remodelling, the effects of fondaparinux on human osteoblasts were analysed in vitro. METHODS: Primary human osteoblast cell cultures were incubated with either the low molecular weight heparin dalteparin at concentrations of 30, 300 and 900 microg/ml or with fondaparinux at concentrations of 25, 50, 100, 150, 200 and 250 microg/ml. Cellular proliferation rate and protein synthesis were measured. Expression of genes encoding osteocalcin, collagen type I and alkaline phosphatase was examined by reverse transcriptase-polymerase chain reaction. RESULTS: Incubation with dalteparin led to a significant, dose-dependent inhibition of osteoblast proliferation, inhibition of protein synthesis, and inhibited expression of phenotype markers (osteocalcin and alkaline phosphatase genes) after 3 and 7 days. No inhibitory effects were observed in the fondaparinux-treated cells. CONCLUSION: Fondaparinux did not inhibit osteoblast proliferation in vitro and may reduce the risk of heparin-induced osteoporosis associated with long-term heparin administration.

[Response of osteoblast cultures to titanium, steel and hydroxyapatite implants]. / Verhalten von Osteoblastenkulturen auf Titan-, Stahl- und Hydroxylapatit-Implantaten.

The biocompatibility of bone implants and substitutes is usually tested on cell-lines only, despite a different and more relevant behaviour on primary osteoblasts as final targets can be expected. "Osteoblast-like" cell line (MC3T3-E1) and fresh human osteoblasts (HOB), cultured from cancellous bone grafts from the iliac crest were used for the study. Three different clinically used biomaterials were compared regarding biocompatibility: titanium, steel and hydroxyapatite (Bio-Oss). "Osteoblast-like" cell line and fresh human osteoblasts (5x104) were seeded on the three bone implants. Cell proliferation and osteocalcin synthesis were determined 1, 3, 7, and 10 days after the cells were plated on the biomaterials. All experiments were performed in five times (pro culture double measurements). HOB proliferation on hydroxyapatite was decreasing after 3 days, whereas cells from "Osteoblast-like" cell line showed comparable proliferation to the control group. The most interesting observation was the significant decrease of the osteocalcin levels (in conditioned medium) of "osteoblast like" cells and HOB on HA (Bio-Oss). We conclude that HA disturbs the proliferation and osteocalcin synthesis of HOB.

[Testing bone implants in cell lines and human osteoblasts]. / Testung von Knochenimplantaten auf Zellinien und humanen Osteoblasten.

Two different types of cell-line and fresh human osteoblasts, cultured from cancellous bone grafts from the iliac crest, were used for the study. Three different biomaterials were compared regarding biocompatibility: titanium, steel and hydroxyapatite. The cells were fibroblast cell line (3T3), "osteoblast-like" cell line (MC3T3-E1), and fresh human osteoblasts (HOB) which we cultured in our laboratory. 5 x 10(4) cells of each type were seeded on the three different bone implants. All experiments were performed in triplicate. Cell proliferation and alkaline phosphatase activity were determined 24 and 72 h after the cells were plated on the biomaterials. Human osteoblast growth was better on titanium than on steel and hydroxyapatite. The most remarkable observation was the continuously decreasing alkaline phosphatase activity (ALP) of "osteoblast-like" cells (MC3T3-E1) and human osteoblasts (HOB) on hydroxyapatite. In conclusion, our in vitro observations suggest that the best cell/material interactions were with human osteoblasts (HOB) and titanium.

[Roentgenographic pulmonary findings in long term ventilated multiple trauma patients (author's transl)]. / Röntgenologische Lungenveränderungen bei polytraumatisierten Patienten während Dauerbeatmung.

An analysis of roentgenographic pulmonary findings was done in 46 multiple trauma patients on controlled long term ventilation. 67% of the patients sustained thoracic trauma. The most common lung changes were interstitial and intraalveolar pulmonary edema and inflammatory pneumonia related to contusions, atelectasis and aspiration. The pulmonary lesions predominantly occurred in the middle and lower portions of each lung. The thoracic trauma group had four times as many lung changes than the non-thoracic trauma group. Daily roentgenographic controls are necessary in multiple trauma patients to explain a deterioration in pulmonary gas exchange leading to respiratory failure. To demonstrate discrete lesions in lung parenchyma, we recommend the high voltage technique for chest X-rays in intensive care units.

[Treatment of the polytraumatic shock (author's transl)]. / Behandlung des polytraumatischen Schocks.

The survival rate of patients with multiple injuries can be improved by means of the modern possibilities of rescue services. The timespan preceding treatment can be shortened. A consequent symptomatic treatment of hemodynamic and respiratory disorders plays an important role. A causal therapy of the impaired permeability of the capillaries, which causes the shock, is possible only to some extent. The therapy should concentrate on the solution resulting from shock.

[Haemodynamic, gas exchange and lung X-rays in multiple injuries with blunt chest trauma (author's transl)]. / Hämodynamik, Gasaustausch und radiologische Lungenbefunde bei Mehrfachverletzten mit stumpfem Thoraxtrauma.

In a controlled prospective study of 50 multiple trauma patients special cardio-respiratory monitoring was performed from the moment of admission to the hospital up to seven days. We compared haemodynamic and respiratory data and roentgenograms from 34 patients with blunt thoracic trauma with those of 16 patients without thoracic trauma. The mortality of the thoracic trauma group was higher (56%) than in the control group (19%). Long term intubation was necessary in the first group for an average of 17 days and in the second group for 6.2 days. In the thoracic trauma group there was a higher total pulmonary resistance and a lower cardiac index. Despite mechanical ventilation the pulmonary gas exchange was impaired in the thoracic trauma cases, especially due to an increased shunt fraction. In 16 of the thoracic-trauma patients roentgenograms detected at least temporary pulmonary oedema simultaneously with the increase of shunt fraction and the impairment of pulmonary gas exchange of those patients.

[Hemodynamics and pulmonary gas exchange in radiographically observed posttraumatic lung edema (author's transl)]. / Hämodynamik und pulmonaler Gasaustausch bei radiologisch beobachteten posttraumatischen Lungenödemen.

A prospective study of 46 multiple trauma patients was undertaken using standard intensive care techniques with special circulatory and respiratory monitoring. In 19 cases roentgenograms detected at least temporary pulmonary edema; only 3 of these patients sustained no discernible lung contusions. The hemodynamic data did not explain the pulmonary edema. Despite ventilatory support of all patients, pulmonary gas exchange was abnormal prior to radiological findings of pulmonary edema. The most significant explanation of the investigated data is early capillary leakage in the lung inducing an enlarged transit space at the interstitial site.

[Extravascular lung water in traumatic shock of the dog (author's transl)]. / Das extravasculäre Lungenwasser im traumatischen Schock beim Hund.

Fourteen dogs underwent standardized traumatic hemorrhagic shock. Periodically, measurement of extravascular lung water (EVLW) was taken by thermo-green-dye dilution technique and related to microvascular pressure (MVP) values. An initial drop in EVLW, corresponding to MVP was seen. During shock time the EVLW rose again, while the MVP remained at low levels. This shows a permeability leak during shock time.

[Misinterpretation of chest roentgenogram in intensive care patients (author's transl)]. / Fehlermöglichkeiten bei der Interpretation der Röntgenthoraxaufnahme in der Intensivmedizin.

Misinterpretation of chest roentgenograms in intensive care patients can be largely obviated by a skilled technique and a thorough knowledge on the part of the radiologist of the structural changes in the lungs and their differential diagnostic significance. Correct interpretation is facilitated by relating the observed changes to the pathophysiological clinical findings.

Coagulation and fibrinolysis in multiple trauma after early heparinizing.

In 27 multiple trauma patients receiving standard shock management and intensive care, coagulation and fibrinolysis were investigated after early heparinization. The general coagulation tests did not imply any impaired clotting function. Platelets and factors I, II, and V decreased without induction of hypocoagulability. There was considerable decrease in plasminogen, whereas FDP ranged within normal; thus, a hyperplasminemia can be excluded. Antithrombin III remained within normal range; even in nonsurvivors there was no depletion, although their antithrombin III activity was significantly lower. In comparison to 50 trauma patients - a comparable group with regard to trauma patterns, shock management, and intensive care - there were no significant differences in volume requirements or mortality rate. Whether early heparinization is effective in preventing disseminated intravascular coagulation (DIC) related organ failure remains to be seen.

Early cardiorespiratory prognostic patterns in multiple trauma patients.

Hemodynamic parameters were monitored for one week ( in greatest detail during the first 24 hours) in 50 severely injured people who had been admitted to the hospital for treatment within one hour after the accident and, as judged by the type of injury, had probably lost more than 1 1/2 liters of blood. The estimations included not only blood pressure, heart rate (HR) and shock index but also the cardiac index (CI), pulmonary arterial pressure (PAP), pulmonary capillary pressure (PCP), and parameters of oxygen transport were determined with the aid of Swan-Ganz catheters, CI, SI, and left ventricular stroke work index (LVSWI) were decreased, PAP, total pulmonary resistance (TPR), and total systemic resistance (TSR), were increased from the very beginning in the nonsurviving group, with a high prediction of vital outcome. The Swan-Ganz catheter is valuable in such cases in obtaining early prognostic evaluation and in controlling therapy - ie, volume replacement, vasodilation, and adrenergic therapy.

Cardiorespiratory and metabolic patterns in multiple trauma patients.

During the last 3 years we have performed a detailed study in 50 patients using the Swan-Ganz catheter to provide prognostic haemodynamic and metabolic values at an early stage. There was a total of 320 severe injuries in these 50 patients with a statistical mean of six to seven. The severity of injuries is documented by the volume replacement necessary within the first 24 hr. On average more than 8 litres of whole blood, albumin and dextran were given intravenously. Out of 50 patients, 28 survived. During the first 2h there was a significant difference between survivor and non-survivor in systolic blood pressure and shock index. From the very beginning the surviving patients demonstrated a higher cardiac index than non-survivors. Pulmonary artery pressure and total pulmonary resistance were also elevated from the very beginning. The right ventricular stroke work index was increased; the left ventricular stroke work index was decreased in the group of non-survivors. The arteriovenous oxygen content difference as well as oxygen extraction ratio was elevated, and the oxygen availability was decreased in the group of the non-surviving patients. Of the metabolic parameters, an increase in the lactate/pyruvate ratio as well as in serum glucose and a decrease in base excess values permit an early prediction of a favourable outcome. There were significant differences in the enzymes SGOT, SGPT, LDH, CHE, as well as bilirubin concentrations between survivors and non-survivors from sixth day on.

[Predictive value of metabolic profiles in multiple trauma]. / Prädiktiver Wert metabolischer Profile bei Polytraumen.

An increase in the lactate:pyruvate ratio as well as in serum gluocse levels and a decrease in base excess values permit an early prediction of vital outcome in severely traumatized patients. Further laboratory data such as enzymes, bilirubin, and amino-acid concentrations, however, demonstrate significant differences between survivors and nonsurvivros at a later stage, i.e., from the 6th day on.

[Monitoring of cardio-pulmonary parameters in cases of severe multiple injuries (author's transl)]. / Monitoring kardiopulmonaler Parameter nach schwerem Polytrauma.

Haemodynamic parameters were monitored for one week (in greatest detail during the first 24 hours) in 35 severely injured persons who had been admitted for treatment within one hour after the accident and, as judged by the type of injury, had probably lost more than 1 1/2 litres of blood. The estimations included not only blood pressure, heart rate and shock index; but the cardiac index, pulmonary arterial pressure, pulmonary capillary pressure and parameters of oxygen transport were also determined with the aid of Swan-Ganz catheters. The importance of the data as guide-lines for the correct type of intensive therapy and their prognostic significance are discussed. The evidence provided by the cardiac index, pulmonary arterial and capillary pressure and the parameters of oxygen transport are of particular importance in these cases. A list is added of the values of some haemodynamic factors which should be taken into account in the initial treatment of severely injured persons.

[Clinical and biometric studies of the adaptive response of shock parameters in patients with multiple injuries and its prognostic significance]. / Klinische und biometrische Untersuchungen über das Regelungsverhalten von Schockparametern bei Mehrfachverletzten und ihre prognostische Bedeutung.

In 35 patients with multiple injuries hemodynamic and renal function parameters measured early after trauma were analysed by biometric methods. Statistically significant prognostic results and differences in the cybernetic behaviour could be demonstrated.

[Cardiovascular parameters following severe multiple injuries]. / Herz-Kreislauf-Parameter nach schweren Polytrauma.

Cardiovascular parameters of 35 patients with multiple injuries are monitored over a period of 7 days. Pulmonary artery pressure, pulmonary vascular resistance, intrapulmonary shunts and cardiac output appear to have a significant prognostic value.