Relevance of angiogenesis in autoimmune testis inflammation.

Experimental autoimmune orchitis (EAO) is a useful model to study organ-specific autoimmunity and chronic testicular inflammation. This model reflects testicular pathological changes reported in immunological infertility in men. Progression of EAO in rodents is associated with a significantly increased percentage of testicular endothelial cells and interstitial testicular blood vessels, indicating an ongoing angiogenic process. Vascular endothelial growth factor A (VEGFA), the main regulator of physiological and pathological angiogenesis, can stimulate endothelial cell proliferation, chemotaxis and vascular permeability. The aim of this study was to explore the role of VEGFA in the pathogenesis of testicular inflammation. Our results found VEGFA expression in Leydig cells, endothelial cells and macrophages in testis of rats with autoimmune orchitis. VEGFA level was significantly higher in testicular fluid and serum of rats at the end of the immunization period, preceding testicular damage. VEGF receptor (VEGFR) 1 is expressed mainly in testicular endothelial cells, whereas VEGFR2 was detected in germ cells and vascular smooth muscle cells. Both receptors were expressed in testicular interstitial cells. VEGFR2 increased after the immunization period in the testicular interstitium and VEGFR1 was downregulated in EAO testis. In-vivo-specific VEGFA inhibition by Bevacizumab prevented the increase in blood vessel number and reduced EAO incidence and severity. Our results unveil relevance of VEGFA-VEGFR axis during orchitis development, suggesting that VEGFA might be an early marker of testicular inflammation and Bevacizumab a therapeutic tool for treatment of testicular inflammation associated with subfertility and infertility.

Histamine H4 receptor as a novel therapeutic target for the treatment of Leydig-cell tumours in prepubertal boys.

Leydig-cell tumours (LCTs) are rare endocrine tumours of the testicular interstitium, with recent increased incidence. Symptoms include precocious puberty in children; and erectile dysfunction, infertility and/or gynaecomastia, in adults. So far, scientific evidence points to aromatase (CYP19) overexpression and excessive oestrogen and insulin-like growth factor (IGF) -1 production as responsible for Leydig-cell tumourigenesis. LCTs are usually benign; however, malignant LCTs respond poorly to chemo/radiotherapy, highlighting the need to identify novel targets for treatment. Herein, we investigated the potential role of the histamine receptor H4 (HRH4) as a therapeutic target for LCTs using R2C rat Leydig tumour cells, a well-documented in vitro model for Leydigioma. Also, we studied for the first time the expression of CYP19, IGF-1R, oestrogen receptor (ER) α, ERß, androgen receptor (AR) and HRH4 in human prepubertal LCTs versus normal prepubertal testes (NPTs). HRH4 agonist treatment inhibited steroidogenesis and proliferation in R2C cells and also negatively affected their pro-angiogenic capacity in vitro and in vivo, as assessed by evaluating the proliferative activity of human umbilical vein endothelial cells and by means of the quail chorioallantoic membrane assay, respectively. Moreover, E2 and IGF-1 inhibited HRH4 mRNA and protein levels. In human prepubertal LCTs, CYP19, IGF-1R, ERα and ERß were overexpressed compared with NPTs. In contrast, HRH4 staining was weak in LCTs, but moderate/strong and confined to the interstitium in NPTs. Importantly, HRH4 was absent or barely detectable in seminiferous tubules or germ cells. Overall, our results point to HRH4 as a novel therapeutic target in LCTs.

In Vitro Human Umbilical Vein Endothelial Cells Response to Ionic Dissolution Products from Lithium-Containing 45S5 Bioactive Glass.

Since lithium (Li⁺) plays roles in angiogenesis, the localized and controlled release of Li⁺ ions from bioactive glasses (BGs) represents a promising alternative therapy for the regeneration and repair of tissues with a high degree of vascularization. Here, microparticles from a base 45S5 BG composition containing (wt %) 45% SiO2, 24.5% Na2O, 24.5% CaO, and 6% P2O5, in which Na2O was partially substituted by 5% Li2O (45S5.5Li), were obtained. The results demonstrate that human umbilical vein endothelial cells (HUVECs) have greater migratory and proliferative response and ability to form tubules in vitro after stimulation with the ionic dissolution products (IDPs) of the 45S5.5Li BG. The results also show the activation of the canonical Wnt/ß-catenin pathway and the increase in expression of proangiogenic cytokines insulin like growth factor 1 (IGF1) and transforming growth factor beta (TGFß). We conclude that the IDPs of 45S5.5Li BG would act as useful inorganic agents to improve tissue repair and regeneration, ultimately stimulating HUVECs behavior in the absence of exogenous growth factors.

Efecto terapéutico del extracto etanólico de las hojas de Oenothera rosea A. "Chupasangre", en forma de crema farmacéutica / Therapeutic effect of the ethanolic extract of the leaves of Oenothera rosea A. "Chupasangre", in the form of a pharmaceutical cream

El presente trabajo de investigación tuvo como objetivo evaluar el efecto antiinflamatorio y cicatrizante de la crema farmacéutica a base del extracto etanólico de las hojas de Oenothera rosea "chupasangre" procedente del Departamento de Ancash (Huaraz). Se determinaron los metabolitos secundarios mediante marcha fitoquímica (flavonoides, alcaloides, taninos, saponinas, fenoles, glucósidos y otros). Se evaluó el efecto contra la inflamación y su actividad en las cicatrices en 3 grupos poblacionales (contusiones leves, contusiones moderadas y heridas leves cerradas) de 20 a 50 años de edad, de ambos sexos, los cuales se subdividieron en grupos experimentales y controles, en el Centro de Salud Ganimedes DISA IV LIMA ESTE ­ MINSA del distrito de San Juan de Lurigancho. Se evaluó el estado general para un diagnóstico médico; para luego iniciar el uso tópico por medio de controles de observación y medición de la zona afectada hasta su completa recuperación. Los datos fueron procesados mediante el análisis (ANOVA), Tukey y análisis de varianza, dándonos como resultado que las cremas al 3 y 5 % mostraron buen efecto antiinflamatorio (contusiones leves y contusiones moderadas) y regular efecto cicatrizante (heridas leves cerradas), mientras, que la crema al 1 % no tiene efecto. Además, la crema al 5 % fue sometida a estabilidad acelerada a una temperatura de 40 °C durante 90 días teniendo como parámetros los análisis organolépticos (aspecto, color y olor), fisicoquímicos (pH, viscosidad) y carga microbiológica total; obteniendo como resultado una crema estable y que cumple con los criterios de aceptación.

Nanocomposite scaffolds with tunable mechanical and degradation capabilities: co-delivery of bioactive agents for bone tissue engineering.

Novel multifunctional nanocomposite scaffolds made of nanobioactive glass and alginate crosslinked with therapeutic ions such as calcium and copper were developed for delivering therapeutic agents, in a highly controlled and sustainable manner, for bone tissue engineering. Alendronate, a well-known antiresorptive agent, was formulated into microspheres under optimized conditions and effectively loaded within the novel multifunctional scaffolds with a high encapsulation percentage. The size of the cation used for the alginate crosslinking impacted directly on porosity and viscoelastic properties, and thus, on the degradation rate and the release profile of copper, calcium and alendronate. According to this, even though highly porous structures were created with suitable pore sizes for cell ingrowth and vascularization in both cases, copper-crosslinked scaffolds showed higher values of porosity, elastic modulus, degradation rate and the amount of copper and alendronate released, when compared with calcium-crosslinked scaffolds. In addition, in all cases, the scaffolds showed bioactivity and mechanical properties close to the endogenous trabecular bone tissue in terms of viscoelasticity. Furthermore, the scaffolds showed osteogenic and angiogenic properties on bone and endothelial cells, respectively, and the extracts of the biomaterials used promoted the formation of blood vessels in an ex vivo model. These new bioactive nanocomposite scaffolds represent an exciting new class of therapeutic cell delivery carrier with tunable mechanical and degradation properties; potentially useful in the controlled and sustainable delivery of therapeutic agents with active roles in bone formation and angiogenesis, as well as in the support of cell proliferation and osteogenesis for bone tissue engineering.

Inhibition of angiopoietin-1 (ANGPT1) affects vascular integrity in ovarian hyperstimulation syndrome (OHSS).

Ovarian hyperstimulation syndrome (OHSS) is a complication of ovarian stimulation with gonadotrophins following human chorionic gonadotrophin (hCG) administration. The relationship between hCG and OHSS is partly mediated via the production of angiogenic factors, such as vascular endothelial growth factor A (VEGFA) and angiopoietins (ANGPTs). Here, we investigated the effect of ANGPT1 inhibition on ovarian angiogenesis in follicular fluid (FF) from women at risk of OHSS, using the chorioallantoic membrane (CAM) of quail embryos as an experimental model. We also analysed cytoskeletal changes and endothelial junction protein expression induced by this FF in the presence or absence of an ANGPT1-neutralising antibody in endothelial cell cultures. The presence of this antibody restored the number of vascular branch points and integrin αvß3 levels in the CAMs to control values. ANGPT1 inhibition in FF from OHSS patients also restored the levels of claudin-5, vascular endothelial cadherin and phosphorylated ß-catenin and partially reversed actin redistribution in endothelial cells. Our findings suggest that ANGPT1 increases pathophysiological angiogenesis in patients at risk of OHSS by acting on tight and adherens junction proteins. Elucidating the mechanisms by which ANGPT1 regulates vascular development and cell-cell junctions in OHSS will contribute to identifying new therapeutic targets for the treatment of human diseases with aberrant vascular leakage.

Angiogenic effects of ionic dissolution products released from a boron-doped 45S5 bioactive glass.

In regenerative medicine of vascularized tissues, there is a great interest in the use of biomaterials that are able to stimulate angiogenesis, a process necessary for rapid revascularization to allow the transport and exchange of oxygen, nutrients, growth factors and cells that take part in tissue repair and/or regeneration. An increasing number of publications have shown that bioactive glasses stimulate angiogenesis. Because it has been established that boron (B) may play a role in angiogenesis, the aim of this study was to assess the in vivo angiogenic effects of the ionic dissolution products that from a bioactive glass (BG) in the 45S5 system doped with 2 wt% B2O3 (45S5.2B). The pro-angiogenic capacity of 45S5.2B BG was assessed on the vasculature of the embryonic quail chorioallantoic membrane (CAM). Ionic dissolution products from 45S5.2B BG increased angiogenesis. This is quantitatively evidenced by the greater expression of integrin αvß3 and higher vascular density in the embryonic quail CAM. The response observed at 2 and 5 days post-treatment was equivalent to that achieved by applying 10 µg mL-1 of basic fibroblast growth factor. These results show that the ionic dissolution products released from the bioactive glass 45S5.2B stimulate angiogenesis in vivo. The effects observed are attributed to the presence the ionic dissolution products, which contained 160 ± 10 µM borate.

In vitro endothelial cell response to ionic dissolution products from boron-doped bioactive glass in the SiO2-CaO-P2O5-Na2O system.

As it has been established that boron (B) may perform functions in angiogenesis and osteogenesis, the controlled and localized release of B ions from bioactive glasses (BGs) is expected to provide a promising therapeutic alternative for regenerative medicine of vascularized tissues, such as bone. The aim of this study was to assess the in vitro angiogenic effects of the ionic dissolution products (IDPs) from BGs in the SiO2-CaO-Na2O-P2O5 (45S5) system and of those from 45S5 BG doped with 2 wt% B2O3 (45S5.2B). The results show, for the first time, the IDPs from 45S5.2B BG stimulated human umbilical vein endothelial cell (HUVEC) proliferation and migration that were associated with phosphorylation of extracellular signal-related kinase (ERK) 1/2, focal adhesion kinase (FAK) and p38 protein. It was also shown that IDPs from 45S5.2B BG could enhance in vitro HUVEC tubule formation and secretion of interleukin 6 (IL6) and the basic fibroblast growth factor (bFGF). The effects observed are attributed to the presence of B in the IDPs. These findings are relevant to bone tissue engineering and regeneration because the IDPs from 45S5.2B BG may act as inexpensive inorganic angiogenic agents providing a convenient alternative to the application of conventional angiogenic growth factors.

Effect of nano-sized bioactive glass particles on the angiogenic properties of collagen based composites.

Angiogenesis is essential for tissue regeneration and repair. A growing body of evidence shows that the use of bioactive glasses (BG) in biomaterial-based tissue engineering (TE) strategies may improve angiogenesis and induce increased vascularization in TE constructs. This work investigated the effect of adding nano-sized BG particles (n-BG) on the angiogenic properties of bovine type I collagen/n-BG composites. Nano-sized (20-30 nm) BG particles of nominally 45S5 Bioglass® composition were used to prepare composite films, which were characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The in vivo angiogenic response was evaluated using the quail chorioallantoic membrane (CAM) as an model of angiogenesis. At 24 h post-implantation, 10 wt% n-BG containing collagen films stimulated angiogenesis by increasing by 41 % the number of blood vessels branch points. In contrast, composite films containing 20 wt% n-BG were found to inhibit angiogenesis. This experimental study provides the first evidence that addition of a limited concentration of n-BG (10 wt%) to collagen films induces an early angiogenic response making selected collagen/n-BG composites attractive matrices for tissue engineering and regenerative medicine.

Short esophagus: selection of patients for surgery and long-term results.

BACKGROUND: After improvements in antireflux surgery (ARS), a percentage of reherniations still has cause of failure attributed to a reopening of the hiatal closure or to an untreated short esophagus. However, the existence of short esophagus and its treatment results still are matters of debate. METHODS: The consecutive medical records containing prospective collective data for patients with gastroesophageal reflux disease (GERD) during the period 2001-2009 were analyzed retrospectively. Every patient considered to be a candidate for ARS was studied with a dynamic contrast radiologic study (DCRxS) in which the esophageal length was evaluated. The choice of surgical technique takes into account the motility status of the esophagus and its estimated length. In the postoperative period, every patient had a DCRxS and an endoscopy 1 year after surgery and then after 3 years. Satisfaction with the procedure was surveyed. RESULTS: The consecutive medical records of 437 GERD patients showed that 171 underwent ARS. During the preoperative DCRxS, a short esophagus was suspected in 26 patients. A short esophagus was confirmed for 11 patients (6.4% of the surgically treated patients), and a Collis procedure plus a funduplication was performed. At the preoperative endoscopy, two patients had a normal mucosa, four patients had esophagitis, and five patients had Barrett's esophagus (BE). In the postoperative period, seven patients presented with a healthy mucosa, one BE had disappeared, and the remaining four BEs remained unchanged. During an average follow-up period of 43 months, no reherniations occurred. The 11 patients achieved good symptoms control and would choose surgery again. CONCLUSIONS: Short esophagus can be suspected during preoperative studies, and in this series, it was confirmed in 6.4% of the patients who had surgery. A Collis fundoplication procedure seems to be an adequate operation to control reflux symptoms and to avoid reherniation over the long-term follow-up period.

Histomorphometric and microchemical characterization of maturing dental enamel in rats fed a boron-deficient diet.

Few reports are available in the literature on enamel formation under nutritional deficiencies. Thus, we performed a study to determine the effects of boron (B) deficiency on the maturing dental enamel, employing the rat continuously erupting incisor as the experimental model. Male Wistar rats, 21 days old, were used throughout. They were divided into two groups, each containing ten animals: +B (adequate; 3-mg B/kg diet) and -B (boron deficient; 0.07-mg B/kg diet). The animals were maintained on their respective diets for 14 days and then euthanized. The mandibles were resected, fixed, and processed for embedding in paraffin and/or methyl methacrylate. Oriented histological sections of the continuously erupting incisor were obtained at the level of the mesial root of the first molar, allowing access to the maturation zone of the developing enamel. Dietary treatment did not affect food intake and body weight. Histomorphometric evaluation using undecalcified sections showed a reduction in enamel thickness (hypoplasia), whereas microchemical characterization by energy-dispersive X-ray spectrometry did not reveal alterations in enamel mineralization.

Retraction and triangulation with neodymium magnetic forceps for single-port laparoscopic cholecystectomy.

INTRODUCTION: There have been attempts to minimize the invasiveness of laparoscopic cholecystectomy by reducing the size and/or the number of the operating ports and instruments. These attempts create technical challenges related principally to retraction and triangulation necessary to expose the surgical field for a safe surgery. A new technique based on retraction and triangulation with magnetic instruments for single port laparoscopic surgery is presented. METHODS: Between March 2007 and December 2008, 40 laparoscopic cholecystectomies were performed with single-port laparoscopic surgery with the assistance of magnetic forceps (IMANLAP project). The surgical technique is described, and the intraoperative and postoperative course of the patients is assessed. RESULTS: There were no intraoperative complications, no need to convert to open surgery, and no need to add a second port. Depending on the patient's anatomy, a 1-mm needle was added in some cases. There were no interactions observed between the magnetic devices and the anesthetic monitoring and the rest of the devices of the operation room. CONCLUSIONS: This new procedure is feasible and safe. The main goal is control of the magnetic field, allowing enough controlled strength for retraction and sufficient triangulation for adequate exposure of the surgical field. This allows for the use of a single port through which an optic device with a working channel can perform the operation with safety. Finally, the procedure can be performed in a manner similar to the traditional laparoscopic cholecystectomy, and it also appears to be simple to learn.

Empleo de prótesis expandibles metálicas en el tubo digestivo / Use of metalic expandible prothesis in digestive tract

Se presenta la experiencia en el empleo de prótesis expandibles metálicas en patología neoplásica del tubo digestivo. Entre enero de 1993 y agosto de 1996 colocamos 35 prótesis expandibles metálicas en 33 pacientes, la edad promedio fue de 76 años (50-94). Veintiseis pacientes se presentaron con disfagia, uno con fístula traqueoesofágica con neumopatía y seis con obstrucción intestinal por estenosis colorrectal. Observamos 33 por ciento de complicaciones mayores; hemorragia, migración de la prótesis y oclusión.La mortalidad dentro de los 30 días fue del 7,4 por ciento. En las estenosis altas se obtuvo una adecuada paliación de la disfagia y en las estenosis colorrectales permitió resolver la oclusión intestinal como tratamiento definitivo o prequirúrgico, evitando las intervenciones complejas de urgencia y favoreciendo operaciones en un tiempo. Futuros estudios prospectivos deberán evaluar el costo beneficio de estos nuevos procedimientos

Empleo de prótesis expandibles metálicas en el tubo digestivo / Use of metalic expandible prothesis in digestive tract

Se presenta la experiencia en el empleo de prótesis expandibles metálicas en patología neoplásica del tubo digestivo. Entre enero de 1993 y agosto de 1996 colocamos 35 prótesis expandibles metálicas en 33 pacientes, la edad promedio fue de 76 años (50-94). Veintiseis pacientes se presentaron con disfagia, uno con fístula traqueoesofágica con neumopatía y seis con obstrucción intestinal por estenosis colorrectal. Observamos 33 por ciento de complicaciones mayores; hemorragia, migración de la prótesis y oclusión.La mortalidad dentro de los 30 días fue del 7,4 por ciento. En las estenosis altas se obtuvo una adecuada paliación de la disfagia y en las estenosis colorrectales permitió resolver la oclusión intestinal como tratamiento definitivo o prequirúrgico, evitando las intervenciones complejas de urgencia y favoreciendo operaciones en un tiempo. Futuros estudios prospectivos deberán evaluar el costo beneficio de estos nuevos procedimientos (AU)