Abstracts of the 21 st Congress of the Spanish Society for Surgical Research , Madrid , October 2015 ADIPOSE MESENCHYMAL STEM CELLS IN THE MANAGEMENT OF HIND LIMB CHRONIC ISCHAEMIA

s of the 21st Congress of the Spanish Society for Surgical Research, Madrid, October 2015 ADIPOSE MESENCHYMAL STEM CELLS IN THE MANAGEMENT OF HIND LIMB CHRONIC ISCHAEMIA B. Herrero de la Parte1, I. García-Alonso1, A. González Bada1, M.D. García-Vázquez2, M.C. Morales2 1Dpt of Surgery, Faculty of Medicine, University of The Basque Country (UPV/EHU). 2BioCruces Health Research Institute, Barakaldo, Spain. Background: Current treatments for chronic hind limb ischaemia benefit just 30% of all the patients, with a 25% failure rate. It has been proposed that adipose mesenchymal stem cells (AMCS) could improve revascularization of ischaemich tissues. Methods: In 20 athymic mice (Hsd:Athymic Nude-Foxn1nu) 5 mm of the femoral artery were removed. Half of the animals received 6*105 AMCS in three punctures of 50 μl in the semimembranous muscle. The other 10 animals received the same amount of saline (150 μl). Capillary blood flow was assessed by laser Doppler (Moor Instruments, Devon, UK) both in the ischaemic and the normal limb (7th, 14th and 21st days). After sacrifice, gastrocnemius muscle was removed and the number of capillary vessels was assessed. Results: After 7 days, blood flow in control limbs was reduced to 25% while in treated limbs it was 35% (p< 0.05). After 14 days, the figures were 43% vs 61% (p< 0.05), without further recovery on day 21st. In treated muscles regenerating muscle fibres could be found on 21st day, while in control muscles only neutrophil infiltration with some grade of necrosis could be observed. Capillary count showed 25 in treated muscles against only 16 in control samples. Conclusions: AMCS inoculated into ischaemic muscle increases vascular restoration (both capillary blood flow and capillary counts). ANESTHETIC EFFECT IN MECHANICAL CIRCULATORY ASSIST DEVICES. EXPERIMENTAL STUDY B. Gómez-Tapia, P. Morillas-Sendin, M. Barranco, JF Del Cañizo, B. Quintana-Villamandos. Dpt. of Anaesthesiology & Intensive Care, Gregorio Marañon University General Hospital, Madrid, Spain Objectives: The main purpose of ventricular assist devices (VAD) is to maintain cardiac output and thus the perfusion of vital organs. Nowadays, the VAD can act as a bridge to transplantation, as a bridge to recovery and as definitive therapy. The aim of our study was to compare the effect of sevoflurane and propofol in organ blood flow in a porcine model with a VAD. Methods: 10 minipigs were divided into two groups depending on the anaesthetic used: sevoflurane (SEVO group, n= 5) and propofol (PROP group, n= 5). A centrifugal pump Biomedicus 540 was implanted; the output cannula of the device was anastomosed to the ascending aorta, and the input cannula was placed through the apex of the left ventricle. To measure organ blood flow, we used coloured microspheres injected through the left atrium before assistance (basal moment) and after 30 minutes of partial support. Once the experience was finished, the animal was sacrificed and biopsies of brain, heart, liver, lung and kidney were taken to measure organ blood flow. Results: Blood flow was significantly higher in the brain (frontal lobes), heart (right and left ventricle), and liver after 30 minutes in the SEVO group, although no significant differences were recorded for the lung, and kidneys. Conclusions: In our study, sevoflurane showed higher blood flow in the brain, heart and liver compared with propofol after implantation of a continuous-flow VAD in an experimental porcine model. If these results were confirmed through clinical trials, sevoflurane would be the optimal option for anaesthetic maintenance for patients with a VAD. ANTIPROLIFERATIVE EFFECT OF HYPERTHERMIA & CHEMOTHERAPY ON CC-531 COLORECTAL CANCER CELLS IN VITRO B. Herrero de la Parte1, D. Sáenz de Urturi1, J.J. Echevarría-Uraga2, C. Acebal Montero1, A. González Bada1, A. Santamaría López1, I. García-Alonso1 1Dpt of Surgery, Faculty of Medicine, University of The Basque Country (UPV/EHU).2 Dpt. of Radiology, Hospital de Galdakao-Usánsolo. Spain. Background: Despite increasingly better results of hepatic surgery for colorectal liver metastases, new therapies are needed to reduce/avoid recurrence. Hyperthermia has been proposed as a possible complement to increase the efficiency of adjuvant chemotherapy. This experience was designed to check whether hyperthermia modifies cell response to chemotherapy. Methods: All-Trans-Retinoic Acid 10 μM (ATRA), 5-Fluorouracil 0.25 μg/ml (5-FU) were added alone or combined to cultures of CC-531 cells. After 24, 48 or 72 hours cultures were placed in an incubator at 47 ∘C for 45’. Cell viability was assessed by MTT just after hyperthermia and 24, 48 and 72 hours later. Data were analysed with ANOVA (p< 0.05 regarded as significant). Results: Cultures treated with drugs showed the deepest drop at 24 h (ATRA 45%, 5-FU 30%, ATRA&5-FU 34%). Following hyperthermia, cell counts were progressively reduced, dropping to just 13% of the initial count after 48 h. When hyperthermia and chemotherapy were combined, cell counts continued falling for 24 h more, with a mean count of 5% at 72 h, if compared to non treated cultures. If we analyse the effect of adding chemotherapy to cells exposed to hyperthermia, it may be observed that cell numbers are halved (ATRA 48%, 5-FU 47%, ATRA&5-FU 42%). Conclusions: Hyperthermia has significantly reduced colorectal cancer cell proliferation in vitro. Chemotherapy prior to hyperthermia has avoided proliferative restart seen after 48 hours. BIOCHEMICAL EVALUATION OF FOLIC ACID TREATMENT OF ISCHAEMIA-REPERFUSION INJURY IN LOWER LIMBS, AFTER TOURNIQUET USE I. Cearra1, T. Chaoui-El-Kaid2, F. Aguayo2, F. Silio1, B. Herrero de la Parte3, I. Garcia-Alonso3 1Dpt of Traumatology and Orthopaedic Surgery, 2Clinical Chemistry Laboratory; Basurto University Hospital. 3Laboratory of Experimental Surgery, University of the Basque Country (UPV/EHU), Spain. Background: There is increasing evidence about the clinical relevance of the injury following tourniquet use in extremity surgery, especially in muscular tissue. This injury is mostly attributable to the so called ischaemia-reperfusion syndrome. Up to date, there is not any established prophylactic treatment. This experiment was designed to evaluate the effect of Folic Acid as antioxidant in the treatment of lower limb ischaemia-reperfusion injury. Methods: Using a self-developed device, we subjected 12 WAG male rats to a 3 hours non-invasive right lower limb ischemia. Half of the animals (n= 6) did not receive any treatment and were used as controls. The other half (n= 6) received Folic Acid (Folidan® 2,5 mg/kg i.p.) starting 20 minutes before reperfusion. After 3 hours of reperfusion, blood samples were collected. Na, K, Cl, Urea, Creatinine, CK, LD, ALP, ALT and AST were measured. Student’s T test was made. Results: CK was significantly decreased in the Folic group (6456± 1662 UI/l vs 9459± 1467 UI/l; p< 0.01). LD was also significantly decreased in the treated group (832± 204 UI/l vs 1395± 262 UI/l; p< 0.01). There were no relevant changes in the other blood parameters. Conclusions: Folic Acid minimizes the increase of CK and LD after 3 hours lower limb ischaemia. © 2016 The Authors BJS 2016; 103 (S1): 1–10 BJS © 2016 BJS Society Ltd