Mimicking the temperature gradient between the sow's oviduct and uterus improves in vitro embryo culture output.

This work was designed to determine temperature conditions within the reproductive tract of the female pig and study their impact on ARTs. Temperatures were recorded using a laparo-endoscopic single-site surgery assisted approach and a miniaturized probe. Sows and gilts were used to address natural cycle and ovarian stimulation treatments, respectively. According to in vivo values, IVF was performed at three temperature conditions (37.0°C, 38.5°C and 39.5°C) and presumptive zygotes were cultured in these conditions for 20 h, while further embryo culture (EC) (21-168 h post-insemination) was maintained at 38.5°C. After 20 h, different fertility parameters were assessed. During EC, cleavage and blastocyst stages were evaluated. Sperm membrane fluidity at the experimental temperatures was studied by using differential scanning calorimetry and fluorescence recovery after photobleaching techniques. An increasing temperature gradient of 1.5°C was found between the oviduct and uterus of sows (P < 0.05) and when this gradient was transferred to pig in vitro culture, the number of poly-nuclear zygotes after IVF was reduced and the percentage of blastocysts was increased. Moreover, the temperature transition phase for the boar sperm membrane (37.0°C) coincided with the temperature registered in the sow oviduct, and sperm membranes were more fluid at 37.0°C compared with those of sperm incubated at higher temperatures (38.5°C and 39.5°C). These data suggest that there may be an impact of physiological temperature gradients on human embryo development.

Mimicking physiological O2 tension in the female reproductive tract improves assisted reproduction outcomes in pig.

STUDY QUESTION: Is O2 tension in the pig oviduct and uterus affected by the estrous cycle stage and the animal's age, and can the outcome of in vitro embryo development be improved by mimicking these physiological values? SUMMARY ANSWER: O2 tension within the pig reproductive organs is affected by the animal's age, and values close to those measured in vivo have a positive impact on embryo development and quality when used during IVF and embryo culture (EC). WHAT IS KNOWN ALREADY: To obtain a healthy embryo in vitro, it is necessary to adopt a culture microenvironment that approximates physiological conditions. Despite advances in surgical procedures and sensitive probes that allow accurate assessment of in vivo O2 tension, few such studies have been conducted recently in mammals. In addition, no reference values of physiological O2 tension in the reproductive tract exist for large animal models such as pig, and the effect of O2 tension on ART outcomes is unknown. STUDY DESIGN, SIZE, DURATION: This study was conducted in pigs. We measured oviductal and uterine O2 tension (n = 29 and 13, respectively) and then examined how the use of the physiological values in pig IVF and EC affected pig ART output (n = 1447 oocytes). PARTICIPANTS/MATERIALS, SETTING, METHODS: The oviductal and uterine O2 tension at the different stages of the estrous cycle was monitored using a laparo-endoscopic single-site surgery (LESS) assisted approach along with a flexible and thin miniaturized luminescent probe. Two groups of pigs, Large-white × Landrace breed, were used: for the first group, 16 pre-pubertal gilts (5 months old and 95 kg) were induced to ovulate with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG); in the second group 13 mature sows (24-48 months and 185 kg) were used. IVF and EC were performed at two different O2 tensions: Atmospheric O2 (20%) and the mean in vivo value measured (7%). At 18-20 h post-insemination (hpi), a small sample of presumptive zygotes were fixed, stained and examined under epifluorescence microscopy to assess the fertilization rates. At 48 hpi, cleavage was evaluated under the stereomicroscope. Finally, at 180 hpi, development to the blastocyst stage was quantified, blastocyst morphology was assessed, and embryos were fixed and stained to count the mean cell number per blastocyst. MAIN RESULTS AND THE ROLE OF CHANCE: The mean O2 content within the pig oviduct and uterus was always lower than in ambient air. The average O2 percentage was higher in gilts (10.0%) than in sows (7.6%) (P < 0.0001). The cleavage rate of porcine in vitro fertilized embryos maintained under 7% O2 during IVF and EC was significantly higher (60.0 ± 2.3) compared with those cultured under 20% O2 (32.0 ± 2.2) (P < 0.05). An increase in the number of cells in embryos cultured under the low O2 concentration (88.9 ± 5.9) was observed compared to those cultured under 20% O2 (59.0 ± 5.0) (P < 0.05). LARGE SCALE DATA: None. LIMITATIONS, REASONS FOR CAUTION: Although minimally invasive surgery was used the effect of anesthesia and manipulations on O2 tension within the organs are unknown. WIDER IMPLICATIONS OF THE FINDINGS: Using physiological oxygen concentrations in IVF/EC could improve ART outcomes. STUDY FUNDING AND COMPETING INTEREST(S): This study was funded by Spanish Ministry of Economy and Competitiveness (MINECO) and European Regional Development Fund (FEDER). Grants AGL2012-40180-C03-01 and AGL2015-66341-R. The authors declare no conflict of interest.

Oocyte holding in the Iberian red deer (Cervus elaphus hispanicus): Effect of initial oocyte quality and epidermal growth factor addition on in vitro maturation.

Current in vitro embryo production protocols in the Iberian red deer (Cervus elaphus hispanicus) need to be optimized; oocyte harvesting in situ followed by overnight holding could reduce the human effort and shipping costs. In our work, post-mortem ovaries were retrieved, and the oocytes were harvested and allocated to G1 group (good quality) or G2 + G3 group (low quality). The oocytes were separately subjected to immediate in vitro maturation (IVM) or held overnight in a holding medium composed of 40% of TCM 199 with Earle's salts, 40% TCM 199 with Hanks' salts and 20% fetal bovine serum (FBS), at room temperature (16 hr). In vitro maturation was carried out in a basal medium supplemented or not with 50 ng/ml of epidermal growth factor (EGF). Our data showed that addition of EGF to the maturation medium increases the percentage of G1 oocytes reaching metaphase II (3.9% vs. 50%, basal vs. EGF; p < .001) and decreased their degeneration rate (69.9% vs. 22.2%, basal vs. EGF; p < .01) when oocytes were immediately matured. Overnight holding increased the meiotic competence of G1 oocytes (37.5% matured in basal medium) and EGF increased prophase arrest in G2 + G3 oocytes (16.1% vs. 38.8% in germinal vesicle [GV] stage in basal medium vs. EGF added medium; p < .05). Our data demonstrate that oocyte holding can be used in Iberian red deer oocytes. Interestingly, EGF addition increases the oocytes' meiotic competence in immediately matured oocytes but not after oocyte holding depending upon initial oocyte quality.

Comparison of minilaparoscopy and single-incision intragastric surgery for gastric submucosal tumor resection in a porcine model.

BACKGROUND: The aim of this study was to develop an easy-to-induce and reproducible model of gastric submucosal tumor in swine to compare minilaparoscopy (ML) with single-incision (SI) intragastric surgery. METHODS: Twelve healthy female pigs (weight 30.94 ± 2.49 kg) underwent a transparietal injection of sterile alginate at the level of Z-line (n = 6) and at the pre-pyloric area (n = 6) creating a model of gastric submucosal pseudotumor. The operative procedures included intragastric resection with ML and SI approaches of cardiac and pre-pyloric lesions, with gastroscopic assistance. After resection, the gastric mucosal layer was closed using intracorporeal sutures. The operative time, complication rate and clinical evolution after 1 month were compared in the four groups that the pigs were arranged. RESULTS: The pseudotumors ranged in size from 3 to 6 cm in diameter. The access of the gastric cavity and resection of the experimental SMP and suturing of the mucosa were performed successfully in 12 animals using both approaches. Mean time to perform the exeresis of gastric cardia tumors was significantly higher in single-incision approach. No significant differences were observed in the surgical time during pyloric surgery. Minilaparoscopic approach reduced significantly the mucosa closure time in esophagogastric and pyloric pseudotumors. One month after, no alterations were shown in the abdominal cavity using exploratory laparotomy. CONCLUSIONS: The technical feasibility of performing safe and efficient intragastric approach of submucosal pseudotumors in swine model was verified in this study. Intragastric ML has advantages over SI, namely regarding the reduction in total surgical times and the fewer technical difficulties.

Objective analysis of surgeons' ergonomy during laparoendoscopic single-site surgery through the use of surface electromyography and a motion capture data glove.

BACKGROUND: Adding to the ergonomic inconveniences already presented by traditional laparoscopy (LAP), laparoendoscopic single-site (LESS) surgery has been found to entail other more specific problems, including greater reduction in movement freedom, in-line vision with loss of triangulation, and greater proximity of instruments. The objective of this study was to evaluate surgeons' ergonomy during LESS surgery, through the study of muscular activity, wrist angle, and hand movements, and compare it with conventional laparoscopy. METHODS: The study group was composed by 14 experienced laparoscopic surgeons, all right-handed. Each one performed dissection tasks on a physical simulator through LAP and LESS approaches. For LAP, straight laparoscopic scissors and dissector were used, whilst for LESS articulating tip scissors and dissector were chosen. During both tasks, muscular activity of biceps brachii, triceps brachii, forearm flexors and extensors, and trapezius muscles was registered through surface electromyography. Simultaneously right-hand movements and wrist angles were obtained through a motion capture data glove (CyberGlove(®)), which allowed for the use of a modified RULA test applied to the recorded angles with subsequent establishment of risk levels for the wrist joint. RESULTS: Muscular activity for trapezius (LAP 6.94 ± 4.12 vs. LESS 11.32 ± 4.68; p ≤ 0.05) and forearm extensor muscles (LAP 9.2 ± 2.45 vs. LESS 37.07 ≤ 16.05; p ≤ 0.001) was significantly lower in conventional laparoscopy compared with LESS approach. No statistical significance was obtained between the different sensors, except in 3 of the 11 analyzed CyberGlove(®) sensors. The modified RULA test showed a score of 3 for laparoscopy (unacceptable), whereas for LESS a score of 2 was obtained (acceptable), with statistically significant differences between them (p ≤ 0.05). CONCLUSIONS: The LESS approach entails greater level of muscular activity in the trapezius and forearm extensor muscles, but we have found evidences of a better wrist position during LESS compared with traditional laparoscopy.

Osteogenic-angiogenic coupled response of cobalt-containing mesoporous bioactive glasses in vivo.

The incorporation of cobalt ions into the composition of bioactive glasses has emerged as a strategy of interest for bone regeneration purposes. In the present work, we have designed a set of bioactive mesoporous glasses SiO2-CaO-P2O5-CoO (Co-MBGs) with different amounts of cobalt. The physicochemical changes introduced by the Co2+ ion, the in vitro effects of Co-MBGs on preosteoblasts and endothelial cells and their in vivo behaviour using them as bone grafts in a sheep model were studied. The results show that Co2+ ions neither destroy mesoporous ordering nor inhibit in vitro bioactive behaviour, exerting a dual role as network former and modifier for CoO concentrations above 3 % mol. On the other hand, the activity of Co-MBGs on MC3T3-E1 preosteoblasts and HUVEC vascular endothelial cells is dependent on the concentration of CoO present in the glass. For low Co-MBGs concentrations (1mg/ml) cell viability is not affected, while the expression of osteogenic (ALP, RUNX2 and OC) and angiogenic (VEGF) genes is stimulated. For Co-MBGs concentration of 5 mg/ml, cell viability decreases as a function of the CoO content. In vivo studies show that the incorporation of Co2+ ions to the MBGs improves the bone regeneration activity of these materials, despite the deleterious effect that this ion has on bone-forming cells for any of the Co-MBG compositions studied. This contradictory effect is explained by the marked increase in angiogenesis that takes place inside the bone defect, leading to an angiogenesis-osteogenesis coupling that compensates for the partial decrease in osteoblast cells. STATEMENT OF SIGNIFICANCE: The development of new bone grafts implies to address the need for osteogenesis-angiogenesis coupling that allows bone regeneration with viable tissue in the long term. In this sense the incorporation of cobalt ions into the composition of bioactive glasses has emerged as a strategy of great interest in this field. Due to the potential cytotoxic effect of cobalt ions, there is an important controversy regarding the suitability of their incorporation in bone grafts. In this work, we address this controversy after the implantation of cobalt-doped mesoporous bioactive glasses in a sheep model. The incorporation of cobalt ions in bioactive glasses improves the bone regeneration ability of these bone grafts, due to enhancement of the angiogenesis-osteogenesis coupling.

[Design and validation of a training model on paediatric and neonatal surgery]. / Diseño y validación de un programa formativo en cirugía laparoscópica pediátrica y neonatal.

We present our experience in the design and development of a training program in paediatric and neonatal laparoscopic surgery, and the determination of face validity by the attendants. Data included in the present study was obtained from five consecutive editions of our Neonatal and Paediatric Laparoscopic Surgery Course. Our training model, with a total duration of 21 hours, begins with acquisition of knowledge in ergonomics and instrument concepts, after which the attendants develop basic laparoscopic dexterity through the performance of hands-on physical simulator tasks. During the second and third days of the course, surgeons undertook various surgical techniques hands-on animal model. At the end of the training program, a subjective evaluation questionnaire was handed out to the attendants, in which different didactic and organizational aspects were considered. We obtained a highly positive score on all questions concerning the different topics and techniques included in the training program (> or = 9 points over 10). 78,5% of the 54 attendants was in accordance with the course total duration, whilst 21,5% considered that it should be of longer duration. Regarding abilities' self assessment, 79,1% considered themselves capacitated to perform trained procedures on live patients. The presented training model has obtained a very positive valuation score, leading to an increase in the attendants' self confidence in the application of learned techniques to their clinical practice.

Injectable mesoporous bioactive nanoparticles regenerate bone tissue under osteoporosis conditions.

The osteogenic capability of mesoporous bioactive nanoparticles (MBNPs) in the SiO2CaO system has been assessed in vivo using an osteoporotic rabbit model. MBNPs have been prepared using a double template method, resulting in spherical nanoparticles with a porous core-shell structure that has a high surface area and the ability to incorporate the anti-osteoporotic drug ipriflavone. In vitro expression of the pro-inflammatory genes NF-κB1, IL-6, TNF-α, P38 and NOS2 in RAW-264.7 macrophages, indicates that these nanoparticles do not show adverse inflammatory effects. An injectable system has been prepared by suspending MBNPs in a hyaluronic acid-based hydrogel, which has been injected intraosseously into cavitary bone defects in osteoporotic rabbits. The histological analyses evidenced that MBNPs promote bone regeneration with a moderate inflammatory response. The incorporation of ipriflavone into these nanoparticles resulted in a higher presence of osteoblasts and enhanced angiogenesis at the defect site, but without showing significant differences in terms of new bone formation. STATEMENT OF SIGNIFICANCE: Mesoporous bioactive glass nanoparticles have emerged as one of the most interesting materials in the field of bone regeneration therapies. For the first time, injectable mesoporous bioactive nanoparticles have been tested in vivo using an osteoporotic animal model. Our findings evidence that MBG nanoparticles can be loaded with an antiosteoporotic drug, ipriflavone, and incorporated in hyaluronic acid to make up an injectable hydrogel. The incorporation of MBG nanoparticles promotes bone regeneration even under osteoporotic conditions, whereas the presence of IP enhances angiogenesis as well as the presence of osteoblast cells lining in the newly formed bone. The injectable device presented in this work opens new possibilities for the intraosseous treatment of osteoporotic bone using minimally invasive surgery.

Emergency transvaginal hybrid natural orifice transluminal endoscopic surgery.

In a clinical series, 10 consecutive female patients with intra-abdominal infections were successfully treated with natural orifice transluminal endoscopic surgery (NOTES) performed transvaginally. The surgery, which consisted of a hybrid NOTES procedure using a transvaginal approach, was performed on an emergency basis by the surgical team on call. The indications for surgery were acute cholecystitis (n = 6), acute appendicitis (n = 2), and pelvic peritonitis (n = 2) with intra-abdominal infection. The procedure was successfully performed in all patients using a dual-channel endoscope and mini-laparoscopy assistance. This is the first clinical series in which NOTES has been performed on an emergency basis to treat intra-abdominal infections. Transvaginal surgery for intra-abdominal infection is a feasible procedure for groups experienced in the elective NOTES approach.

Anatomical changes due to pneumoperitoneum analyzed by MRI: an experimental study in pigs.

PURPOSE: Different effects on cardiovascular and respiratory systems and liver are associated with pneumoperitoneum. This study aimed to determine the morphological changes in the abdominal anatomy as a result of increased intra-abdominal pressure due to pneumoperitoneum using MRI. METHODS: Ten healthy female pigs were used in this study. MRI studies of the abdomen in supine position were made before the creation of pneumoperitoneum and 1 h after increasing the pressure to 14 mmHg. Changes in area, volume, and longitudinal and transverse length of the liver were measured. The diameters of the lumen of the abdominal aorta, the inferior vena cava and portal vein were observed in three positions along the abdominal cavity. The position of the diaphragm after the induction of pneumoperitoneum was also analyzed. RESULTS: After induction of pneumoperitoneum, volume and transverse length of the liver was significantly increased, while peak area was decreased. Stenosis in the aortic lumen was observed (P < 0.05). Longitudinal and transverse diameters of the portal lumen were reduced, but significant differences were only found in the longitudinal diameter. Alterations in the diameter of the inferior vena cava lumen were obtained in three analyzed positions, but differences were significant only in two of them. A mean cranial displacement of the diaphragm equal to 25 mm was also observed. CONCLUSION: Increasing abdominal pressure up to laparoscopic pressure (14 mmHg) provokes morphological changes in the liver, vascular structures and diaphragm. These changes could be related to functional alterations that different organs experience after the induction of pneumoperitoneum.

[Proposed experimental model to provide training in the management of anesthesia in liver transplantation]. / Propuesta de un programa de formación usando un modelo experimental para el manejo anestésico del trasplante hepático.

OBJECTIVE: To propose and evaluate the use of a porcine model for training in how to manage and improve the quality of anesthesia during liver transplantation. MATERIAL AND METHODS: Five trained anesthesiologists who had no previous experience in managing anesthesia during liver transplantation and who did not work in hospitals where the procedure was performed volunteered for the training course and evaluated it. Each trainee anesthetized 5 animals. Surgery was performed under total balanced anesthesia with monitoring of hemodynamics, ventilation, biochemistry, arterial blood gases, and coagulation. Previously set criteria were used to evaluate the trainees' skill in maintaining patient stability. Their work was assessed 7 times: at baseline, 15 minutes into the preanhepatic phase, at the end of the preanhepatic stage, 15 minutes into the anhepatic phase, at the end of the anhepatic phase, 15 minutes into the postanhepatic phase, and at the end of the postanhepatic phase. After completing the course, the trainees filled in a questionnaire to evaluate its usefulness. Analysis of variance was applied to score changes in anesthetic quality criteria. RESULTS: After implantation, changes in metabolic (acidosis) and cardiac (hypotension and bradycardia) status were the most critical abnormalities the trainees faced. Their skill in coping with hemodynamic and metabolic changes improved significantly (P<.05). All participants expressed a preference for receiving training in a porcine model before providing anesthesia in this surgical setting. CONCLUSIONS: Under this training model, the anesthesiologists understood and were able to manage the hemodynamic and physiologic changes that develop during a liver transplant procedure. We believe that training using a porcine model allows an anesthesiologist to acquire experience and skill in this setting.

Development and assessment of an ex-vivo bench model aimed at laparoscopic ureteric reconstructive techniques.

INTRODUCTION/BACKGROUND: Reconstructive and ablative urologic techniques require special technical mastery, especially the intracorporeal suturing. OBJECTIVE: To report the subjective evaluation of a versatile ex-vivo model aimed to practice laparoscopic ureteric reconstructive techniques (LURT) on box-trainer. STUDY DESIGN: The model is a continuous portion of porcine urinary bladder ("dilated pelvis"), the vesico-ureteral joint ("stenosis") and healthy ureter. All 127 participants (n = 119 urologists and n = 8 paediatric surgeons) performed on the model laparoscopic Anderson-Hynes dismembered pyeloplasty, and then, in the animal model, different LURT procedures (ureteroneocystostomy, ureteric reimplantation and/or dismembered pyeloplasty). The model was subjectively evaluated (face and content validity), through a 12 items questionnaire, based on a Likert scale (1-5 points) and a global question (1-10 points). RESULTS: The total mean rating for 11/12 items was very high (>4points). Only one was rated under 3 points. The overall total mean rating from 1 to 10 points was very high (9.19 ± 0.82 points). In 10/12 items, expert's feedback (content validity) prevailed over non-experts (face validity). DISCUSSION/CONCLUSION: The model was highly accepted for the practice of LURT techniques. Additionally, it is cost-effective, easy to assemble, ethically considerate, and realistic.

Silicon substituted hydroxyapatite/VEGF scaffolds stimulate bone regeneration in osteoporotic sheep.

Silicon-substituted hydroxyapatite (SiHA) macroporous scaffolds have been prepared by robocasting. In order to optimize their bone regeneration properties, we have manufactured these scaffolds presenting different microstructures: nanocrystalline and crystalline. Moreover, their surfaces have been decorated with vascular endothelial growth factor (VEGF) to evaluate the potential coupling between vascularization and bone regeneration. In vitro cell culture tests evidence that nanocrystalline SiHA hinders pre-osteblast proliferation, whereas the presence of VEGF enhances the biological functions of both endothelial cells and pre-osteoblasts. The bone regeneration capability has been evaluated using an osteoporotic sheep model. In vivo observations strongly correlate with in vitro cell culture tests. Those scaffolds made of nanocrystalline SiHA were colonized by fibrous tissue, promoted inflammatory response and fostered osteoclast recruitment. These observations discard nanocystalline SiHA as a suitable material for bone regeneration purposes. On the contrary, those scaffolds made of crystalline SiHA and decorated with VEGF exhibited bone regeneration properties, with high ossification degree, thicker trabeculae and higher presence of osteoblasts and blood vessels. Considering these results, macroporous scaffolds made of SiHA and decorated with VEGF are suitable bone grafts for regeneration purposes, even in adverse pathological scenarios such as osteoporosis. STATEMENT OF SIGNIFICANCE: For the first time, the in vivo behavior of scaffolds made of silicon substituted hydroxyapatites (SiHA) has been evaluated under osteoporosis conditions. In order to optimize the bone regeneration properties of these bioceramics, 3D macroporous scaffolds have been manufactured by robocasting and implanted in osteoporotic sheep. Our experimental design shed light on the important issue of the biological response of nano-sized bioceramics vs highly crystalline bioceramics, as well as on the importance of coupling vascularization and bone growth processes by decorating SiHA scaffolds with vascular endothelial growth factor.

Meshes in a mess: Mesenchymal stem cell-based therapies for soft tissue reinforcement.

Surgical meshes are frequently used for the treatment of abdominal hernias, pelvic organ prolapse, and stress urinary incontinence. Though these meshes are designed for tissue reinforcement, many complications have been reported. Both differentiated cell- and mesenchymal stem cell-based therapies have become attractive tools to improve their biocompatibility and tissue integration, minimizing adverse inflammatory reactions. However, current studies are highly heterogeneous, making it difficult to establish comparisons between cell types or cell coating methodologies. Moreover, only a few studies have been performed in clinically relevant animal models, leading to contradictory results. Finally, a thorough understanding of the biological mechanisms of mesenchymal stem cells in the context of foreign body reaction is lacking. This review aims to summarize in vitro and in vivo studies involving the use of differentiated and mesenchymal stem cells in combination with surgical meshes. According to preclinical and clinical studies and considering the therapeutic potential of mesenchymal stem cells, it is expected that these cells will become valuable tools in the treatment of pathologies requiring tissue reinforcement. STATEMENT OF SIGNIFICANCE: The implantation of surgical meshes is the standard procedure to reinforce tissue defects such as hernias. However, an adverse inflammatory response secondary to this implantation is frequently observed, leading to a strong discomfort and chronic pain in the patients. In many cases, an additional surgical intervention is needed to remove the mesh. Both differentiated cell- and stem cell-based therapies have become attractive tools to improve biocompatibility and tissue integration, minimizing adverse inflammatory reactions. However, current studies are incredibly heterogeneous and it is difficult to establish a comparison between cell types or cell coating methodologies. This review aims to summarize in vitro and in vivo studies where differentiated and stem cells have been combined with surgical meshes.

Validation of a new artificial model for simulated training of a laparoscopic vesicourethral anastomosis. / Validación de un nuevo modelo de simulación inorgánica para la anastomosis uretrovesical laparoscópica.

OBJECTIVE: The aim of this study is to prove the effectiveness of a low cost, artificial model for training of a laparoscopic urethrovesical anastomosis. MATERIALS AND METHODS: This study included urologists who attended specialised courses on laparoscopic radical prostatectomy (LRP) held during the period 2015 to 2017. They were divided into 2 groups according to their previous experience in laparoscopic surgery. The tasks performed on the artificial simulator were prostate resection, "task 1", and urethrovesical anastomosis, "task 2". Once these exercises were completed, the study participants filled in an anonymous questionnaire regarding their demographic data and experience level in laparoscopic surgery (LS). In addition, they gave their opinions about the didactic capacity of the artificial organ and evaluated its usefulness as a tool for LRP training. To demonstrate face and content validity, the participants judged the texture, consistency, morphology and evaluated its similarity to the real organ. The assessment was made with a five-point Likert scale. RESULTS: The students were divided into 2groups: 10 experts (Group E) and 12 novices (Group N). The only significant difference between the scores of novices and experts was regarding the inclusion of this tool in the training programs (Group E=5 points versus group N=4.4±0.59, P=.024). The experts' group rated all the items with higher scores than the novices' one. Regarding the general assessment of the simulation model, the novice participants gave an average score of 8.00±0.91 points out of 10, while the experts' group granted higher scores of 9.4±0,51. CONCLUSION: This artificial model has shown to have an elevated face, content and construct validity, as well being an optimal didactic tool for training in the techniques of prostate resection and laparoscopic urethrovesical anastomosis.

Mesoporous bioactive glass/ɛ-polycaprolactone scaffolds promote bone regeneration in osteoporotic sheep.

Macroporous scaffolds made of a SiO2-CaO-P2O5 mesoporous bioactive glass (MBG) and ɛ-polycaprolactone (PCL) have been prepared by robocasting. These scaffolds showed an excellent in vitro biocompatibility in contact with osteoblast like cells (Saos 2) and osteoclasts derived from RAW 264.7 macrophages. In vivo studies were carried out by implantation into cavitary defects drilled in osteoporotic sheep. The scaffolds evidenced excellent bone regeneration properties, promoting new bone formation at both the peripheral and the inner parts of the scaffolds, thick trabeculae, high vascularization and high presence of osteoblasts and osteoclasts. In order to evaluate the effects of the local release of an antiosteoporotic drug, 1% (%wt) of zoledronic acid was incorporated to the scaffolds. The scaffolds loaded with zoledronic acid induced apoptosis in Saos 2 cells, impeded osteoclast differentiation in a time dependent manner and inhibited bone healing, promoting an intense inflammatory response in osteoporotic sheep. STATEMENT OF SIGNIFICANCE: In addition to an increase in bone fragility and susceptibility to fracture, osteoporosis also hinders the clinical success of endosseous implants and grafting materials for the treatment of bone defects. For the first time, macroporous scaffolds made of mesoporous bioactive glass and ε-caprolactone have been evaluated in a sheep model that mimics the osteoporosis conditions in humans. These implants fostered bone regeneration, promoting new bone formation at both the peripheral and the inner parts of the scaffolds, showing thick trabeculae and a high vascularization degree. Our results indicate that macroporous structures containing highly bioactive mesoporous glasses could be excellent candidates for the regenerative treatment of bone defects in osteoporotic patients.

Synergistic effect of Si-hydroxyapatite coating and VEGF adsorption on Ti6Al4V-ELI scaffolds for bone regeneration in an osteoporotic bone environment.

The osteogenic and angiogenic responses to metal macroporous scaffolds coated with silicon substituted hydroxyapatite (SiHA) and decorated with vascular endothelial growth factor (VEGF) have been evaluated in vitro and in vivo. Ti6Al4V-ELI scaffolds were prepared by electron beam melting and subsequently coated with Ca10(PO4)5.6(SiO4)0.4(OH)1.6 following a dip coating method. In vitro studies demonstrated that SiHA stimulates the proliferation of MC3T3-E1 pre-osteoblastic cells, whereas the adsorption of VEGF stimulates the proliferation of EC2 mature endothelial cells. In vivo studies were carried out in an osteoporotic sheep model, evidencing that only the simultaneous presence of both components led to a significant increase of new tissue formation in osteoporotic bone. STATEMENT OF SIGNIFICANCE: Reconstruction of bones after severe trauma or tumors extirpation is one of the most challenging tasks in the field of orthopedic surgery. This scenario is even more complicated in the case of osteoporotic patients, since their bone regeneration capability is decreased. In this work we present a porous implant that promotes bone regeneration even in osteoporotic bone. By coating the implant with osteogenic bioceramics such as silicon substituted hydroxyapatite and subsequent adsorption of vascular endothelial growth factor, these implants stimulate the bone ingrowth when they are implanted in osteoporotic sheep.

Technical feasibility of laparoscopic Finney pyloroplasty examined in a canine model.

The efficacy of laparoscopic surgery in the treatment of chronic duodenal ulcer has been demonstrated using minimally invasive approaches. This study aimed to evaluate the technical feasibility of laparoscopic Finney pyloroplasty in six dogs. Under laparoscopic guidance, the anastomosis was created using mechanical devices (n = 3) and combining a linear stapler device and intracorporeal suturing (n = 3). The operative time and complications were recorded. Evaluation of the anastomosis included studies of intraoperative and postoperative endoscopy and ultrasonography as well as the complete gastric emptying time. The animals were killed 4 weeks after surgery. Measurement of luminal diameter, amount of adhesion formation, degree of healing, and inflammation or fibrosis were evaluated in the postmortem studies. The operation was successfully completed for all the animals. No intraoperative or postoperative complications were observed. Gastric emptying was significantly enhanced in the postoperative period, as compared with the preoperative results. After 1 month, there was no evidence of anastomotic leak dehiscence at the gastroduodenal anastomosis. Luminal diameter was increased, and no abnormal findings were encountered during the postmortem abdominal exploration. The technical feasibility of performing a safe laparoscopic pyloroplasty in a dog model was demonstrated in this study.

Tissue consistency perception in laparoscopy to define the level of fidelity in virtual reality simulation.

BACKGROUND: What degree of fidelity must a laparoscopic simulator have to achieve a training objective? This difficult question is addressed by studying the sensory interaction of surgeons in terms of a surgical skill: tissue consistency perception. METHODS: A method for characterizing surgeon sensory interaction has been defined and applied in an effort to determine the relative importance of three components of perceptual surgical skill: visual cues, haptic information, and previous surgical knowledge and experience. Expert, intermediate, and novel surgeons were enrolled in the study. Users were asked to rank tissue consistency in four different conditions: a description of the tissue alone (Q), visual information alone (VI), tactile information alone (TI), and both visual and tactile information (VTI). Agreement between these stages was assessed by a coefficient of determination (R2). RESULTS: Tissue is a determinant factor (p < 0.001) in the perception of tissue consistency, whereas the expertise of the surgeon is not (p = 0.289). Tissue consistency perception is based mainly on tactile information (TI-VTI agreement is high, R2 = 0.873), although little sensory substitution is present (VI-VTI agreement is low, R2 = 0.509). Agreement of Q-VI increases with experience (R2 = 0.050, 0.290, and 0.573, corresponding with to novel, intermediate, and expert surgeons), which has been associated with the "visual haptics" concept. CONCLUSIONS: Virtual reality simulators need haptic devices with force feedback capability if tissue consistency information is to be delivered. On the other hand, the visual haptics concept has been associated with a kind of tactile memory developed by surgical experience.

[Animal models in urological laparoscopic training]. / Modelos experimentales en la cirugía laparoscópica urológica.

We present the experience of the Minimally Invasive Surgery Centre (MISC) in the development of a modular training model in laparoscopic surgery. The experience analysis includes the description of the training objectives, the learning process of simple and advance laparoscopic urologic techniques, as well as some current and future considerations before applying the laparoscopic techniques. This learning program pretends to optimize the knowledge of the surgeon and the clinical practice of these surgical techniques. The phases of the learning process have been classified in four levels, which include different modules and models and whose application will depend on the experience and surgical skills. This pyramidal training system permits the student to advance step by step through each level depending on her surgical skills. We have presented our experience in twelve courses about laparoscopic urology and four courses of laparoscopic radical prostatectomy, in which more than 300 urologists have assisted. Furthermore, some Spanish Urology Units have been developing special experimental training programs on laparoscopic radical prostatectomy, partial nephrectomy or laparoscopic dismembered pyeloplasty with Anderson-Hynes technique. It has been previously described that laparoscopic modular learning constitutes a very useful concept to avoid problems related to an incomplete and incorrect learning process. Also it seems clear that the laparoscopic training reduces the learning curve in laparoscopic urologic techniques.