Re-endothelialization of punctured ePTFE graft: an in vitro study under pulsed perfusion conditions.

BACKGROUND: When used as arteriovenous (AV) shunts for haemodialysis, small diameter expanded polytetrafluoroethylene (ePTFE) grafts have a high failure rate in vivo. Attempts to improve graft patency are various, and focus on either improvement of implantation techniques or graft tissue engineering. The tissue engineering approach attempts to reproduce in grafts the properties of pristine vasculature. As shown in previous experiments, it is possible to grow on ePTFE grafts under shear stress in vitro an autologous endothelial cell layer, which will withstand physiological stress under in vivo conditions of blood flow. The aim of this study was to investigate in an in vitro model the regenerative potency of a tissue-engineered prosthetic vascular graft after repeated cannulation with a haemodialysis cannula. METHODS: Pig endothelial cells were harvested from an external jugular vein. Following processing of the endothelial cells, seven ePTFE grafts were coated with an inner cell layer and were kept under pulsed perfusion. Each graft was then cannulated three times with a standard shunt needle. The endothelium was then left to regenerate for a maximum of 48 h. The grafts were stained with haematoxylin/eosin before histological study. RESULTS: All grafts were endothelialized over the puncture sites within 48 h. Histological analysis revealed a confluent endothelial cell lining at each puncture site. Cell morphology and cell pattern over puncture sites were not different from randomly picked locations over the graft lumen. CONCLUSION: Our results underline the potential of endothelial tissue engineering in vascular shunt surgery. Vascular bio-hybrids that have the properties of pristine vascular endothelium may be a key step forward in maintaining angio-access in patients who require haemodialysis.

Intraperitoneal administration of the angiogenesis inhibitor thalidomide does not impair anastomotic healing following large bowel resection in a rabbit model.

The effects of thalidomide after intraperitoneal instillation on the healing of colonic anastomoses are not known. A series of 40 New Zealand White rabbits underwent an end-to-end colonic anastomosis. The animals were randomized into four groups. Groups 1 (n = 10) and 2 (n = 10) were treated with dissolved thalidomide 200 mg/kg intraperitoneally, whereas groups 3 (n = 10) and 4 (n = 10) were treated only with the dissolver. Animals were sacrificed at day 3 (groups 1, 3) and day 7 (groups 2, 4). Anastomotic healing was tested by measuring the bursting pressure in vitro. Immunohistochemical staining of the anastomotic site was performed with polyclonal antibodies against CD31 and Mib-1, to determine a possible antiangiogenic or antiproliferative effect. Statistical analysis was performed using Spearman's log rank correlation and paired t-test. On postoperative day 3 (p > 0.19) and postoperative day 7 (p > 0.73), there was no difference in bursting pressure in the treatment and the control groups. Angiogenesis scores were reduced at day 3 in the thalidomide group (p < 0.05), but did not differ between the groups at day 7. White blood cell counts were decreased in the treatment groups at day 3 (p < 0.01) and day 7, compared to control groups (p < 0.01). There was no difference in the expression of Mib-1 in either group at day 3 or day 7. The intraperitoneal administration of thalidomide does not interfere with the healing of colonic anastomosis. Although the angiogenesis score is diminished at day 3, this did not lead to a reduced bursting pressure at day 3 or day 7.

Surgical treatment of fractures of the distal clavicle with polydioxanone suture tension band wiring: an alternative osteosynthesis.

Fracture of the distal clavicle type II (Neer) is an indication for surgical intervention. We report our experience in 12 patients with acute clavicular fractures and operative treatment with polydioxanone suture (PDS) tension band wiring. The patients were assessed 6 and 12 weeks postoperatively by radiological and clinical evaluation and with the Constant Murley score. All 12 patients had an excellent functional result 12 weeks postoperatively. The Constant Murley score was excellent in all patients. The PDS band can be considered as an alternative osteosynthesis. In the context of the current literature, the advantages and disadvantages of this new procedure are discussed.

Serum vascular endothelial growth factor levels correlate better with tumour stage in small cell lung cancer than albumin, neuron-specific enolase or lactate dehydrogenase.

OBJECTIVE: Vascular endothelial growth factor (VEGF) is an important cytokine in the process of angiogenesis. Elevated serum levels of the cytokine may determine cancer patients who will benefit from adjuvant anti-angiogenic therapy in the future. To correlate serum levels of VEGF with tumour stage and established prognostic markers in patients with small cell lung cancer (SCLC), a prospective study was performed on 70 patients. METHODOLOGY: From August 1999 to May 2000, 70 consecutive patients (51 male, 19 female) with histologically proven SCLC were enrolled into the study. Staging of the disease included clinical investigation, bronchoscopy, chest X-ray, thoracic computed tomography and ultrasound. The patients were grouped into five stages according to the Marburg classification (very limited disease (VLD), limited disease (LD), extensive disease I (EDI), extensive disease II (EDII) and extensive disease III (EDIII)). Prior to treatment, a 10 mL serum sample from each patient was examined by ELISA to quantify levels of VEGF and neuron-specific enolase (NSE). Lactate dehydrogenase (LDH) and albumin levels were determined by photomorphometric analysis. Statistical analysis was performed using the Waller-Duncan k ratio t-test and Pearson's correlation test. RESULTS: Serum VEGF levels correlated well with tumour stage (P < 0.0001). Albumin levels were not correlated with tumour stage, but levels of NSE and LDH increased with stage progression. When patients were divided into two groups (VLD and LD vs EDI-III), VEGF levels were significantly lower in the initial stages of the disease compared with extensive disease (P < 0.0001). Serum levels of VEGF correlated better with tumour stage than did concentrations of NSE, LDH or albumin. CONCLUSION: Serum VEGF levels may serve as an additional prognostic marker in the course of patients with SCLC. Further studies are needed to determine whether these patients may benefit from additional anti-angiogenic therapy in the future.

In vivo measurement of oxygen-derived free radicals during reperfusion injury.

By use of an optimized cytochrome c-based biosensor, superoxide radical production was measured continuously in vivo. The aim of this study was the online detection of superoxide concentration during reperfusion after a variable time of ischemia. Measurements were performed by placing the detecting sensor into gastrocnemius muscle tissue. Ischemia was induced by clamping the vena and arteria femoralis. Current response of the sensor was recorded continuously as an equivalent for superoxide concentration. Ischemia times varied from 5 to 120 minutes. The minimum ischemia time to record superoxide production was 10 minutes. By inducing longer periods of ischemia, an increase in superoxide concentration reached its highest levels at 2 hours. Furthermore, the total time of superoxide production after reperfusion depended on the total time of ischemia.

Thalidomide given intraperitoneally reduces the number of postoperative adhesions after large bowel resection in rabbits.

OBJECTIVE: To investigate the effect of thalidomide given intraperitoneally on the formation of adhesions after colonic resection in rabbits. DESIGN: Controlled, randomised prospective study. SETTING: University hospital, Germany. ANIMALS: 40 female New Zealand White rabbits. INTERVENTIONS: After an end-to-end colonic anastomosis the animals were given thalidomide 200 mg/kg in 1% carboxymethylcellulose intraperitoneally or carboxymethylcellulose alone (n = 20 in each group). OUTCOME MEASURES: The adhesion score according to Tyrell on day 3 (n = 20) and day 7 (n = 20) postoperatively, weight, behaviour, and white cell count (WCC). RESULTS: There was no difference in behaviour or weight. On the third postoperative day WCCs and on the seventh postoperative day WCCs and adhesion scores, were lower in the thalidomide group (p < 0.01). CONCLUSION: The number of postoperative adhesions was reduced in the group given thalidomide intraperitoneally. Our results suggest that thalidomide may be helpful in the prevention of postoperative adhesions.