Publications (28)

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    [Show abstract] [Hide abstract] ABSTRACT: Irreversible electroporation is a fast-growing liver ablation technique. Although safety has been well documented in small ablations, our aim is to assess its safety and feasibility when a large portion of liver is ablated. Eighty-seven mice were subjected to high voltage pulses directly delivered across parallel plate electrodes comprising around 40% of mouse liver. One group consisted in 55 athymic-nude, in which a tumor from the KM12C cell line was grown and the other thirty-two C57-Bl6 non-tumoral mice. Both groups were subsequently divided into subsets according to the delivered field strength (1000 V/cm, 2000 V/cm) and whether or not they received anti-hyperkalemia therapy. Early mortality (less than 24 hours post-IRE) in the 2000 V/cm group was observed and revealed considerably higher mean potassium levels. In contrast, the animals subjected to a 2000 V/cm field treated with the anti-hyperkalemia therapy had higher survival rates (OR = 0.1, 95%CI = 0.02-0.32, p < 0.001). Early mortality also depended on the electric field magnitude of the IRE protocol, as mice given 1000 V/cm survived longer than those given 2000 V/cm (OR = 4.7, 95%CI = 1.8-11.8, p = 0.001). Our findings suggest that ionic disturbances, mainly due to potassium alterations, should be warned and envisioned when large volume ablations are performed by IRE.
    Full-text Article · Apr 2016 · Scientific Reports
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    [Show abstract] [Hide abstract] ABSTRACT: Purpose: The aim of this study was to assess the capacity of two methods of surgical pancreatic stump closure in terms of reducing the risk of pancreatic fistula formation (POPF): radiofrequency-induced heating versus mechanical stapler. Materials and methods: Sixteen pigs underwent a laparoscopic transection of the neck of the pancreas. Pancreatic anastomosis was always avoided in order to work with an experimental model prone to POPF. Pancreatic stump closure was conducted either by stapler (ST group, n = 8) or radiofrequency energy (RF group, n = 8). Both groups were compared for incidence of POPF and histopathological alterations of the pancreatic remnant. Results: Six animals (75%) in the ST group and one (14%) in the RF group were diagnosed with POPF (p = 0.019). One animal in the RF group and three animals in the ST group had a pseudocyst in close contact with both pancreas stumps. On day 30 post-operation (PO), almost complete atrophy of the exocrine distal pancreas was observed when the main pancreatic duct was efficiently sealed. Conclusions: Our findings suggest that RF-induced heating is more effective at closing the pancreatic stump than mechanical stapler and leads to the complete atrophy of the distal remnant pancreas.
    Full-text Article · Jan 2016 · International Journal of Hyperthermia
  • [Show abstract] [Hide abstract] ABSTRACT: Background: Pancreatic duct ligation (PDL) has been used as a model of chronic pancreatitis and as a model to increase β-cell mass. However, studies in mice have demonstrated acinar regeneration after PDL, questioning the long-term validity of the model. We aim to elucidate whether RF-assisted transection (RFAT) of the main pancreatic duct is a reliable PDL model, both in short (ST, 1-month) and long-term (LT, 6-months) follow-ups. Methods: Eleven pigs were subjected to RFAT. Biochemical (serum/peripancreatic amylase and glucose) and histological changes (including a semiautomatic morphometric study of over 1000 images/pancreas and IHC analysis) were evaluated after ST or LT follow-up and also in fresh pancreas specimens that were used as controls for 1 (n = 4) and 6 months (n = 6). Results: The distal pancreas in the ST was characterized by areas of acinar-to-ductal metaplasia (56%) which were significantly reduced at LT (21%) by fibrotic replacement and adipose tissue. The endocrine mass showed a normal increase. Conclusion: RFAT in the pig seems to be an appropriate PDL model without restoration of pancreatic drainage or reduction of endocrine mass.
    Article · Jan 2016
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    [Show abstract] [Hide abstract] ABSTRACT: Objective. The aim of this study was to find a proper experimental design and to evaluate n-butyl-2-cyanoacrylate (Histoacryl) as a fixation method for a light-weight and large pore PP mesh (Synthetic PP Mesh-1) using the sheep as an animal model. Methods. Posterior vaginal implantation by means of episiotomy was used to implant 8 ewes which were evaluated macroscopically and histologically at 3 months () and 6 months () post-surgery. In previous pilot studies anterior vaginal implantation was evaluated, as well as different synthetic mesh materials, sizes and fixation methods ( to 3) during three weeks. In all cases a clinical evaluation of the animal was performed. Results. A reduction in the mesh size (Synthetic PP Mesh-1) together with precise application of the surgical glue Histoacryl to fix the mesh yielded significantly better histocompatibility results () compared to larger size or other fixation methods. Conclusion. The combination of Synthetic PP Mesh-1 with Histoacryl offered a high degree of graft integration without vaginal ulceration and a minimal foreign body reaction, being the sheep a proper animal model to test these types of medical devices.
    Full-text Article · Jun 2015 · BioMed Research International
  • [Show abstract] [Hide abstract] ABSTRACT: Background: Pancreatic duct ligation (PDL) has been used as a model of chronic pancreatitis and as a model to increase β-cell mass. However, studies in mice have demonstrated acinar regeneration after PDL, questioning the long-term validity of the model. We aim to elucidate whether RF-assisted transection (RFAT) of the main pancreatic duct is a reliable PDL model, both in short (ST, 1-month) and long-term (LT, 6-months) follow-ups. Methods: Eleven pigs were subjected to RFAT. Biochemical (serum/peripancreatic amylase and glucose) and histological changes (including a semiautomatic morphometric study of over 1000 images/pancreas and IHC analysis) were evaluated after ST or LT follow-up and also in fresh pancreas specimens that were used as controls for 1 (n = 4) and 6 months (n = 6). Results: The distal pancreas in the ST was characterized by areas of acinar-to-ductal metaplasia (56%) which were significantly reduced at LT (21%) by fibrotic replacement and adipose tissue. The endocrine mass showed a normal increase. Conclusion: RFAT in the pig seems to be an appropriate PDL model without restoration of pancreatic drainage or reduction of endocrine mass.
    Article · Jun 2015 · Pancreatology
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    Full-text Dataset · Mar 2015
  • M Caminal · D Peris · C Fonseca · [...] · J Vives
    [Show abstract] [Hide abstract] ABSTRACT: Current developments in tissue engineering strategies for articular cartilage regeneration focus on the design of supportive three-dimensional scaffolds and their use in combination with cells from different sources. The challenge of translating initial successes in small laboratory animals into the clinics involves pilot studies in large animal models, where safety and efficacy should be investigated during prolonged follow-up periods. Here we present, in a single study, the long-term (up to 1 year) effect of biocompatible porous scaffolds non-seeded and seeded with fresh ex vivo expanded autologous progenitor cells that were derived from three different cell sources [cartilage, fat and bone marrow (BM)] in order to evaluate their advantages as cartilage resurfacing agents. An ovine model of critical size osteochondral focal defect was used and the test items were implanted arthroscopically into the knees. Evidence of regeneration of hyaline quality tissue was observed at 6 and 12 months post-treatment with variable success depending on the cell source. Cartilage and BM-derived mesenchymal stromal cells (MSC), but not those derived from fat, resulted in the best quality of new cartilage, as judged qualitatively by magnetic resonance imaging and macroscopic assessment, and by histological quantitative scores. Given the limitations in sourcing cartilage tissue and the risk of donor site morbidity, BM emerges as a preferential source of MSC for novel cartilage resurfacing therapies of osteochondral defects using copolymeric poly-D,L-lactide-co-glycolide scaffolds.
    Article · Jan 2015 · Cytotechnology
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    [Show abstract] [Hide abstract] ABSTRACT: Gene therapy is an attractive tool for the treatment of monogenic disorders, in particular for lysosomal storage diseases (LSD) caused by deficiencies in secretable lysosomal enzymes in which neither full restoration of normal enzymatic activity nor transduction of all affected cells are necessary. However, some LSD such as Mucopolysaccharidosis Type IIIB (MPSIIIB) are challenging because the disease's main target organ is the brain and enzymes do not efficiently cross the blood-brain barrier even if present at very high concentration in circulation. To overcome these limitations, we delivered AAV9 vectors encoding for α-N-acetylglucosaminidase (NAGLU) to the Cerebrospinal Fluid (CSF) of MPSIIIB mice with the disease already detectable at biochemical, histological and functional level. Restoration of enzymatic activity in Central Nervous System (CNS) resulted in normalization of glycosaminoglycan content and lysosomal physiology, resolved neuroinflammation, and restored the pattern of gene expression in brain similar to that of healthy animals. Additionally, transduction of the liver due to passage of vectors to the circulation led to whole-body disease correction. Treated animals also showed reversal of behavioural deficits and extended lifespan. Importantly, when the levels of enzymatic activity were monitored in the CSF of dogs following administration of canine NAGLU-coding vectors to animals that were either naïve or had pre-existing immunity against AAV9, similar levels of activity were achieved, suggesting that CNS efficacy would not be compromised in patients seropositive for AAV9. Our studies provide a strong rationale for the clinical development of this novel therapeutic approach as treatment for MPSIIIB. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
    Full-text Article · Dec 2014 · Human Molecular Genetics
  • [Show abstract] [Hide abstract] ABSTRACT: Oocytes from adult goats have better competence to develop up to blastocyst stage than those from prepubertal females. Previous results in our laboratory showed that oocyte developmental competence in prepubertal goats is positively related to follicle diameter, as oocytes from follicles larger than 3mm have the same competence as adult goat ones, demonstrating that follicular environment affects oocyte competence for embryo development. Other studies concluded that fatty acids (FA) in follicular fluid (FF) play an important role in oocyte and embryo development. The aim of this study was to analyse FF composition regarding FA percentage comparing samples from adult (3 years old, forage diet) or prepubertal (1 month suckling) Murciano-Granadina goats. FF samples were recovered from follicles larger and smaller than 3mm by laparoscopy (adult) or aspiration of ovaries collected at slaughterhouse (prepubertal). For the FA analysis, the Sukhija and Palmquist protocol was used with some modifications. Briefly, 200μL of FF sample was vortexed for 60s with 250μL of toluene and 1mL of HCL (5%) and then warmed in water bath for 1h at 70°C. Subsequently, 1.25mL of K2CO3 (12%) and 500μL of toluene was added, vortexed for 30s, and centrifuged for 5min (3000rpm). Finally the supernatant was recovered and dried with Na2SO4. The extracted samples were maintained at -20°C until gas chromatographic analysis (123-2362, Agilent Technologies Inc., Santa Clara, CA). The results are shown in Table 1. The analysis of FA composition in FF showed significantly higher concentrations of C15:0 (pentadecanoic), C16:0 (palmitic), C17:0 (margaric), C18:0 (stearic), linolenic acid (ALA), and EPA (eicosapentanoic) in adult goats compared to prepubertal goats, and C17:0, C18:0 linoleic acid (LA), and ALA in larger follicles of prepubertal females, showing some coincidences in follicular environment that could explain previous studies. These results are in agreement with those of Matoba et al. (2014; Rep. Fertil. Dev. 26, 337) who showed that FF associated with more competent oocytes had a significantly higher concentration in ALA than FF from incompetent oocytes in cattle.
    Article · Dec 2014 · Reproduction Fertility and Development
  • Conference Paper · Nov 2014
  • Daniel Costa · Marta Leiva · Xavier Moll · [...] · Anna Andaluz
    [Show abstract] [Hide abstract] ABSTRACT: The purpose of this randomised trial was to compare the effects of alfaxalone and propofol on tear production (STT-1), intraocular pressure (IOP) and globe position (GP) in healthy dogs. Fourteen Beagles were randomly divided into two groups; dogs in one group received alfaxalone (3 mg/kg) (n=7) and dogs in the other group received propofol (6 mg/kg) (n=7), both administered intravenously. IOP and GP were evaluated at basal time (Tb) and T2,5,10,15,20,25&30 (minutes after complete drug administration). STT-1 was evaluated at Tb and T10,20&30. STT-1 and IOP results were analysed using analysis of variance and GP was analysed using the likelihood ratio χ(2) test. Dogs in the alfaxalone group showed a significant reduction in STT-1 at T10&20 (P<0.05), while the propofol group did not show statistically significant differences in this parameter over time. Both anaesthetic drugs produced a transient elevation of IOP at T2 (P>0.05), which then decreased (P<0.01). While alfaxalone caused a ventral globe deviation that lasted from T2 to T10 (P<0.05) and was fully recovered at T30, propofol induced a ventral globe deviation from T2 to T5 (P<0.05), being restored at T20. These results suggest that both alfaxalone and propofol can be safely used for intraocular surgery, as they significantly reduce IOP. Furthermore, anaesthetic induction with propofol would be especially recommended for dogs with tear deficiencies.
    Article · Oct 2014 · The Veterinary record
  • [Show abstract] [Hide abstract] ABSTRACT: Regenerative therapies using adult stem cells have attracted great interest in the recent years and offer a promising alternative to current surgical practices. In this report, we evaluated the safety and efficacy of an autologous cell-based treatment of osteoarthritis using mesenchymal stromal cells expanded from bone marrow aspirates that were administered intra-articularly. Ten 2-year old ewes were divided in two groups (for analysis at 6 and 12 months, respectively). Full thickness articular cartilage defects of approximately 60 mm2 were created arthroscopically in the medial femorotibial condyles and a meniscal tear in the anterior horn of the medial meniscus in the 20 hind legs. Intra-articular injection of 4 mL of either treatment (a suspension of cells) or control (same as treatment, without cells) were applied one month after generating a chronic condition similar to human pathology. Animals were monitored radiographically, by MRI and ultrasound scanning; and macroscopic and histological analyses were conducted at 6 and 12 months. Furthermore a full necropsy was performed at 12 months post-treatment. The intra-articular injection of autologous MSC was safe, as judged by the lack of local or systemic adverse effects during the clinical follow-up and by a full necropsy performed at 12 months post-treatment. Evidence of regeneration of articular cartilage and meniscus was case-dependent but statistically significant improvement was found in specific macroscopic and histological parameters. Such parameters included colour, rigidity, cell distribution and hyaline quality of the refill tissue as well as the structure of subchondral bone.
    Article · Sep 2014 · New Biotechnology
  • M Caminal · X Moll · D Codina · [...] · J Vives
    [Show abstract] [Hide abstract] ABSTRACT: Clinical translation of emerging technologies aiming at cartilage resurfacing is hindered by neither the appropriate scaffold design nor the optimal cell source having been defined. Here, critical-sized, chondral-only focal defects were created in sheep and treated with clinical-grade, co-polymeric poly-lactide:polyglycolic acid scaffolds either alone or seeded with 3.3 × 10(6) ± 0.4 × 10(6) autologous bone marrow-derived mesenchymal stromal cells and studied over 12 month follow-up. An untreated group was included for comparison. Second-look arthroscopy performed at 4 months post-treatment evidenced the generation of neocartilage of better quality in those defects treated with cells. However, macroscopic scores in the cell-treated group declined significantly from 7.5 ± 2.3 at 4 months to 3.1 ± 2.6 (p = 0.0098) at 12 months post-treatment, whereas the other two experimental groups remained unaltered during 4-12 month post-treatment. The effectiveness of the cell-based approach proposed in this study is thus restricted to between months 1 and 4 post-treatment.
    Article · Jun 2014 · Biotechnology Letters
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    Xavier Moll · Felix García · Rosa Isabel Ferrer · [...] · Anna Andaluz
    [Show abstract] [Hide abstract] ABSTRACT: The aim of the study was to determine the distribution of different volumes of methylene blue solution injected into the epidural space in anaesthetized pregnant and non-pregnant sheep, to evaluate its cranial distribution and to compare between them. Fifteen pregnant and fifteen non-pregnant sheep were included in the study. Sheep were anaesthetized and received 0.05, 0.1, or 0.2 mL/kg of a lumbosacral epidural solution containing 0.12% methylene blue in 0.9% saline. Thirty minutes after the epidural injection, the ewes were euthanized. The extension of the dye within the epidural space was measured, and the correlation between the volume of the dye injected and the number of stained vertebrae was evaluated. The cranial migration of the dye between pregnant and non-pregnant sheep was also compared. The results show that the volume of methylene blue injected epidurally into pregnant and non-pregnant sheep correlated directly with its cephalic distribution into the epidural space; and a volume of 0.1 mL/kg or 0.2 mL/kg stained up to the first lumbar segment in pregnant and non-pregnant sheep, respectively. Also, the results suggest that the volume of drugs administered into the epidural space of pregnant sheep should be half the volume that would be used in non-pregnant sheep.
    Full-text Article · Apr 2014 · PLoS ONE
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    [Show abstract] [Hide abstract] ABSTRACT: The “One world, one health” initiative emphasizes the need for new strategies to control human and animal tuberculosis (TB) based on their shared interface. A good example would be the development of novel universal vaccines against Mycobacterium tuberculosis complex (MTBC) infection. This study uses the goat model, a natural TB host, to assess the protective effectiveness of a new vaccine candidate in combination with Bacillus Calmette-Guerin (BCG) vaccine. Thirty-three goat kids were divided in three groups: Group 1) vaccinated with BCG (week 0), Group 2) vaccinated with BCG and boosted 8 weeks later with a recombinant adenovirus expressing the MTBC antigens Ag85A, TB10.4, TB9.8 and Acr2 (AdTBF), and Group 3) unvaccinated controls. Later on, an endobronchial challenge with a low dose of M. caprae was performed (week 15). After necropsy (week 28), the pulmonary gross pathology was quantified using high resolution Computed Tomography. Small granulomatous pulmonary lesions (< 0.5 cm diameter) were also evaluated through a comprehensive qualitative histopathological analysis. M. caprae CFU were counted from pulmonary lymph nodes. The AdTBF improved the effects of BCG reducing gross lesion volume and bacterial load, as well as increasing weight gain. The number of Ag85A-specific gamma interferon-producing memory T-cells was identified as a predictor of vaccine efficacy. Specific cellular and humoral responses were measured throughout the 13-week post-challenge period, and correlated with the severity of lesions. Unvaccinated goats exhibited the typical pathological features of active TB in humans and domestic ruminants, while vaccinated goats showed only very small lesions. The data presented in this study indicate that multi-antigenic adenoviral vectored vaccines boosts protection conferred by vaccination with BCG.
    Full-text Article · Nov 2013 · PLoS ONE
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    Anna Andaluz · Laura Santos · Félix García · [...] · Xavier Moll
    [Show abstract] [Hide abstract] ABSTRACT: The objective of this study was to determine the pharmacodynamics effects of the anaesthetic alfaxalone in 2-hydroxypropyl- β -cyclodextrin in pregnant sheep after the intravenous injection of a 2 mg/kg weight dose. Six pregnant Ripollesa sheep, weighing 47.1 ± 4.4 kg, were used. Twenty-four hours after instrumentation, sheep were anaesthetized with intravenous alfaxalone in cyclodextrin. Time to standing from anaesthesia was 30.0 ± 10.81 min. Foetal heart rate increased significantly during the first 5 min after alfaxalone administration. Significant differences were observed in maternal diastolic arterial blood pressure between minute 10 and minutes 90, 120, 150, 180, 210, and 240. Significant differences were observed for foetal systolic arterial blood pressure between 5 and 30 min after alfaxalone administration. Significant differences in foetal pH were detected during the entire study period, whereas maternal pH returned to baseline values by 60 min after alfaxalone administration. The present study indicated that alfaxalone in 2-hydroxypropyl- β -cyclodextrin administered as an intravenous bolus at 2 mg/kg body weight produced minimal adverse effects and an uneventful recovery from anaesthesia in pregnant sheep and their foetus.
    Full-text Article · Oct 2013 · The Scientific World Journal
  • Xavier Moll · Laura Santos · Felix García · Anna Andaluz
    [Show abstract] [Hide abstract] ABSTRACT: Alfaxalone in a 2-hydroxypropyl-β-cyclodextrin (HPCD) formulation is an intravenous (IV) hypnotic agent characterised by the stability of cardiorespiratory effects after a single-bolus administration. The objective of this study was to investigate the cardiovascular, respiratory, and acid-base effects of alfaxalone-HPCD administered during a continuous rate infusion in six Ripollesa sheep. After instrumentation, a 2mg/kg IV bolus of alfaxalone followed by a continuous infusion of 10mg/kg/h was administered to the sheep. Heart rate, arterial blood pressure, respiratory rate and arterial blood gases were recorded. Occasional side effects and time to standing were also noted. No significant changes were observed in arterial blood pressure, but during the infusion and the initial stages of recovery, a significant increase in heart rate occurred during the last 120min of the study. Significant respiratory depression was detected during the infusion period and the first 15min of recovery. This study showed that a constant rate infusion alfaxalone in un-premedicated sheep produced clinically acceptable haemodynamic results and a mild respiratory depression that may require intermittent positive pressure ventilation.
    Article · Oct 2012 · The Veterinary Journal
  • M-D Torres · A Andaluz · F García · [...] · X Moll
    [Show abstract] [Hide abstract] ABSTRACT: The objective of this prospective study was to determine the effects of a single intravenous bolus of alfaxalone in 2-hydroxypropyl-β-cyclodextrin and propofol on the intraocular pressure (IOP) in sheep. Ten Ripollesa sheep with a bodyweight of 48.5 (6.8) kg (mean [sd]) were used in the study. Twenty-four hours before the experimental procedure, a complete ophthalmic examination was performed in all animals. The day of the study, intravenous alfaxalone (2 mg/kg) or propofol (6 mg/kg) was randomly administered in a cross-over design, with a washout period of two weeks. Measurements of IOP, globe position and pupil size were obtained at basal time, before induction (time 0) and at two, five, 10, 15, 20, 30, 45, 60, 90 and 120 minutes after drug administration. Occasional side effects and time to standing were also noted. Intravenous administration of alfaxalone and propofol in sheep resulted in no alteration of IOP. Nevertheless, a decrease in the pupil size was observed in both groups. This present study shows that alfaxalone and propofol, administrated as a single intravenous bolus, are good options for maintaining IOP during anaesthesia in sheep, although marked miosis was observed after administration.
    Article · Mar 2012
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    Xavier Moll · Laura Fresno · Félix García · [...] · Anna Andaluz
    Full-text Article · Sep 2011 · The Veterinary Journal
  • X Moll · F García · A Andaluz
    Article · Jun 2011