In vivo biocompatibility testing of nanoparticle-functionalized alginate-chitosan scaffolds for tissue engineering applications.

Background: There is a strong interest in designing new scaffolds for their potential application in tissue engineering and regenerative medicine. The incorporation of functionalization molecules can lead to the enhancement of scaffold properties, resulting in variations in scaffold compatibility. Therefore, the efficacy of the therapy could be compromised by the foreign body reaction triggered after implantation. Methods: In this study, the biocompatibilities of three scaffolds made from an alginate-chitosan combination and functionalized with gold nanoparticles (AuNp) and alginate-coated gold nanoparticles (AuNp + Alg) were evaluated in a subcutaneous implantation model in Wistar rats. Scaffolds and surrounding tissue were collected at 4-, 7- and 25-day postimplantation and processed for histological analysis and quantification of the expression of genes involved in angiogenesis, macrophage profile, and proinflammatory (IL-1ß and TNFα) and anti-inflammatory (IL-4 and IL-10) cytokines. Results: Histological analysis showed a characteristic foreign body response that resolved 25 days postimplantation. The intensity of the reaction assessed through capsule thickness was similar among groups. Functionalizing the device with AuNp and AuNp + Alg decreased the expression of markers associated with cell death by apoptosis and polymorphonuclear leukocyte recruitment, suggesting increased compatibility with the host tissue. Similarly, the formation of many foreign body giant cells was prevented. Finally, an increased detection of alpha smooth muscle actin was observed, showing the angiogenic properties of the elaborated scaffolds. Conclusion: Our results show that the proposed scaffolds have improved biocompatibility and exhibit promising potential as biomaterials for elaborating tissue engineering constructs.

Embryonic Hyperglycemia Disrupts Myocardial Growth, Morphological Development, and Cellular Organization: An In Vivo Experimental Study.

Hyperglycemia during gestation can disrupt fetal heart development and increase postnatal cardiovascular disease risk. It is therefore imperative to identify early biomarkers of hyperglycemia during gestation-induced fetal heart damage and elucidate the underlying molecular pathomechanisms. Clinical investigations of diabetic adults with heart dysfunction and transgenic mouse studies have revealed that overexpression or increased expression of TNNI3K, a heart-specific kinase that binds troponin cardiac I, may contribute to abnormal cardiac remodeling, ventricular hypertrophy, and heart failure. Optimal heart function also depends on the precise organization of contractile and excitable tissues conferred by intercellular occlusive, adherent, and communicating junctions. The current study evaluated changes in embryonic heart development and the expression levels of sarcomeric proteins (troponin I, desmin, and TNNI3K), junctional proteins, glucose transporter-1, and Ki-67 under fetal hyperglycemia. Stage 22HH Gallus domesticus embryos were randomly divided into two groups: a hyperglycemia (HG) group, in which individual embryos were injected with 30 mmol/L glucose solution every 24 h for 10 days, and a no-treatment (NT) control group, in which individual embryos were injected with physiological saline every 24 h for 10 days (stage 36HH). Embryonic blood glucose, height, and weight, as well as heart size, were measured periodically during treatment, followed by histopathological analysis and estimation of sarcomeric and junctional protein expression by western blotting and immunostaining. Hyperglycemic embryos demonstrated delayed heart maturation, with histopathological analysis revealing reduced left and right ventricular wall thickness (-39% and -35% vs. NT). Immunoexpression levels of TNNI3K and troponin 1 increased (by 37% and 39%, respectively), and desmin immunofluorescence reduced (by 23%). Embryo-fetal hyperglycemia may trigger an increase in the expression levels of TNNI3K and troponin I, as well as dysfunction of occlusive and adherent junctions, ultimately inducing abnormal cardiac remodeling.

Human Heart Morphogenesis: A New Vision Based on In Vivo Labeling and Cell Tracking.

Despite the extensive information available on the different genetic, epigenetic, and molecular features of cardiogenesis, the origin of congenital heart defects remains unknown. Most genetic and molecular studies have been conducted outside the context of the progressive anatomical and histological changes in the embryonic heart, which is one of the reasons for the limited knowledge of the origins of congenital heart diseases. We integrated the findings of descriptive studies on human embryos and experimental studies on chick, rat, and mouse embryos. This research is based on the new dynamic concept of heart development and the existence of two heart fields. The first field corresponds to the straight heart tube, into which splanchnic mesodermal cells from the second heart field are gradually recruited. The overall aim was to create a new vision for the analysis, diagnosis, and regionalized classification of congenital defects of the heart and great arteries. In addition to highlighting the importance of genetic factors in the development of congenital heart disease, this study provides new insights into the composition of the straight heart tube, the processes of twisting and folding, and the fate of the conus in the development of the right ventricle and its outflow tract. The new vision, based on in vivo labeling and cell tracking and enhanced by models such as gastruloids and organoids, has contributed to a better understanding of important errors in cardiac morphogenesis, which may lead to several congenital heart diseases.

Sodium Alginate/Chitosan Scaffolds for Cardiac Tissue Engineering: The Influence of Its Three-Dimensional Material Preparation and the Use of Gold Nanoparticles.

Natural biopolymer scaffolds and conductive nanomaterials have been widely used in cardiac tissue engineering; however, there are still challenges in the scaffold fabrication, which include enhancing nutrient delivery, biocompatibility and properties that favor the growth, maturation and functionality of the generated tissue for therapeutic application. In the present work, different scaffolds prepared with sodium alginate and chitosan (alginate/chitosan) were fabricated with and without the addition of metal nanoparticles and how their fabrication affects cardiomyocyte growth was evaluated. The scaffolds (hydrogels) were dried by freeze drying using calcium gluconate as a crosslinking agent, and two types of metal nanoparticles were incorporated, gold (AuNp) and gold plus sodium alginate (AuNp+Alg). A physicochemical characterization of the scaffolds was carried out by swelling, degradation, permeability and infrared spectroscopy studies. The results show that the scaffolds obtained were highly porous (>90%) and hydrophilic, with swelling percentages of around 3000% and permeability of the order of 1 × 10−8 m2. In addition, the scaffolds proposed favored adhesion and spheroid formation, with cardiac markers expression such as tropomyosin, troponin I and cardiac myosin. The incorporation of AuNp+Alg increased cardiac protein expression and cell proliferation, thus demonstrating their potential use in cardiac tissue engineering.

Sources, Characteristics, and Therapeutic Applications of Mesenchymal Cells in Tissue Engineering.

Tissue engineering (TE) is a therapeutic option within regenerative medicine that allows to mimic the original cell environment and functional organization of the cell types necessary for the recovery or regeneration of damaged tissue using cell sources, scaffolds, and bioreactors. Among the cell sources, the utilization of mesenchymal cells (MSCs) has gained great interest because these multipotent cells are capable of differentiating into diverse tissues, in addition to their self-renewal capacity to maintain their cell population, thus representing a therapeutic alternative for those diseases that can only be controlled with palliative treatments. This review aimed to summarize the state of the art of the main sources of MSCs as well as particular characteristics of each subtype and applications of MSCs in TE in seven different areas (neural, osseous, epithelial, cartilage, osteochondral, muscle, and cardiac) with a systemic revision of advances made in the last 10 years. It was observed that bone marrow-derived MSCs are the principal type of MSCs used in TE, and the most commonly employed techniques for MSCs characterization are immunodetection techniques. Moreover, the utilization of natural biomaterials is higher (41.96%) than that of synthetic biomaterials (18.75%) for the construction of the scaffolds in which cells are seeded. Further, this review shows alternatives of MSCs derived from other tissues and diverse strategies that can improve this area of regenerative medicine.

Antitumor Effects of Natural Compounds Derived from Allium sativum on Neuroblastoma: An Overview.

Garlic (Allium sativum) has been used in alternative medicine to treat several diseases, such as cardiovascular and neurodegenerative diseases, cancer, and hepatic diseases. Several publications have highlighted other features of garlic, including its antibacterial, antioxidative, antihypertensive, and antithrombotic properties. The properties of garlic result from the combination of natural compounds that act synergistically and cause different effects. Some garlic-derived compounds have been studied for the treatment of several types of cancer; however, reports on the effects of garlic on neuroblastoma are scarce. Neuroblastoma is a prevalent childhood tumor for which the search for therapeutic alternatives to improve treatment without affecting the patients' quality of life continues. Garlic-derived compounds hold potential for the treatment of this type of cancer. A review of articles published to date on some garlic compounds and their effect on neuroblastoma was undertaken to comprehend the possible therapeutic role of these compounds. This review aimed to analyze the impact of some garlic compounds on cells derived from neuroblastoma.

Incorporation of the first and second heart fields and prospective fate of the straight heart tube via in vivo labeling of chicken embryos.

Recent discoveries of at least two heart fields and dynamic nature of cardiac development as well as controversies regarding the participation of heart fields in development of different heart structures led us to investigate the dynamics of incorporation of the first and second heart fields and prospective fate of the straight heart tube by labeling chicken embryos in vivo with the fluorescent lipophilic dye DiI. The cephalic and caudal limits of the anterior and posterior segments of the straight heart tube were labeled in two groups of embryos. Labels were tracked along the "C," "S," and "U" loops up to the tetracavitary or mature heart (n = 30 embryos/group; torsion and looping stage). To determine whether the atria and atrioventricular canal are derived from the first heart field the straight heart tube was cultured in vitro and immunodetection of Sox-9 and troponin I was performed to identify the mesenchymal and myocardial lineages respectively. Proliferating cell nuclear antigen (PCNA) immunodetection was used to determine the involvement of cell proliferation in heart tube development during torsion and looping. Embryological constitution of the straight heart tube and heart looping (C, S, and U) were not consistent with current descriptions. In fact, right ventricle precursors were absent in the straight heart tube derived from the first heart field. During torsion and looping, the cephalic segment of the straight heart tube gradually shifted into the heart tube until it was located at the myocardial interventricular septum in the tetracavitary heart. In contrast, the caudal segment of the straight heart tube was elongated and remodeled to become the first heart field derived left ventricle and the proximal part of the ventricular inlets. The ventricular outflows, right ventricle, distal part of the ventricular inlets, and atria developed from the second heart field.

Burkholderia cenocepacia Induces Macropinocytosis to Enter Macrophages.

Burkholderia cenocepacia is an opportunistic pathogen that infects individuals with cystic fibrosis, chronic granulomatous disease, and other immunocompromised states. B. cenocepacia survives in macrophages in membrane-bound vacuoles; however, the mechanism by which B. cenocepacia gains entry into macrophages remains unknown. After macrophage internalization, survival of B. cenocepacia within a bacteria-containing membrane vacuole (BcCV) is associated with its ability to arrest the maturation of the BcCV. In this study, we show that B. cenocepacia induces localized membrane ruffling, macropinocytosis, and macropinosomes-like compartments upon contact with the macrophage. The Type 3 Secretion System (T3SS) of B. cenocepacia contributes to macrophage entry and macropinosome-like compartment formation. These data demonstrate the ability of Burkholderia to enter macrophages through the induction of macropinocytosis.

Validity and reliability of the Mexican resilience measurement scale in families of children with chronic conditions.

BACKGROUND: The resilience to face disease is a process of positive adaptation despite the loss of health. It involves developing vitality and skills to overcome the negative effects of adversity, risks, and vulnerability caused by disease. In Mexico, the Mexican Resilience Measurement Scale (RESI-M) has been validated with a general population and has a five-factor structure. However, this scale does not allow evaluation of resilience in specific subpopulations, such as caregivers. METHOD: This study investigated the psychometric properties of RESI-M in 446 family caregivers of children with chronic diseases. A confirmatory factor analysis (CFA) was performed, internal consistency values were calculated using Cronbach's alpha coefficient, and mean comparisons were determined using t-tests. RESULTS: The expected five-factor model showed an adequate fit with the data based on a maximum likelihood test. The internal consistency for each factor ranged from .76 to .93, and the global internal consistency was .95. No average difference in RESI-M and its factors was found between women and men. CONCLUSION: The RESI-M showed internal consistency and its model of five correlated factors was valid among family caregivers of children with chronic diseases.

Desarrollo temprano de las cavidades cardiacas ventriculares: Importancia de la Neuregulina 1 / Early development of cardiac ventricles: Importance of neuregulin 1

Introducción. Los defectos septales ventriculares graves, asociados con un ventrículo hipoplásico y otro hiperplásico o con la ausencia total del tabique interventricular, suelen ser incompatibles con la vida embrionaria y fetal. Pese a su gravedad, no se conocen sus causas. Por marcaje in vivo en embriones de pollo se confirmó la importancia de las trabéculas en la morfogénesis del tabique interventricular. Por manipulación genética en embriones de ratón y aves se determinó que la ausencia de la Neuregulina 1 (NRG1) o sus receptores, además de provocar escasa diferenciación de los miocitos ventriculares y deficiente formación de las trabéculas, determina la muerte prematura del embrión. Con base en estos antecedentes, el objetivo fue determinar el papel real de NRG1 en la trabeculogénesis temprana y su importancia en la regulación de la proliferación y apoptosis. Métodos. Se estableció un modelo de órgano cultivo de corazón de embrión de pollo previo al inicio de la trabeculogénesis. Se realizaron ensayos de inhibición total de la actividad de NRG1 endógena y posterior adición de la proteína exógena a diferentes concentraciones, determinando la actividad cíclica de los miocitos ventriculares con el antígeno nuclear de proliferación celular y la apoptosis con lisotraker. Resultados. El suero fetal bovino promueve la proliferación, pero impacta negativamente la trabeculogénesis. La adición de NRG1 a concentraciones bajas y períodos cortos de incubación no induce trabeculogénesis. En contraste, a concentraciones medias y períodos de cultivo no mayores a 24 horas, tiene un efecto positivo sobre este proceso. También promueve la proliferación y evita la apoptosis del miocardio ventricular. El incremento en la concentración de NRG1 posiblemente provoca un desbalance molecular que favorece la proliferación desordenada pero no la trabeculogénesis. Conclusiones. El entendimiento del papel de NRG1 en la trabeculogénesis aporta datos para conocer las redes moleculares involucradas también en el desarrollo del tabique interventricular, información indispensable para entender el origen de los defectos septales ventriculares graves.

Background. Serious ventricular septal defects associated with hypoplastic and hyperplasic ventricles or total absence of the interventricular septum (IVS) are usually incompatible with embryonic and fetal life. Despite the importance of these cardiac diseases, their causes are not yet known. Using in vivo labeling in the chick embryo, the importance of ventricular trabeculation was confirmed in IVS morphogenesis. Using knockout mice and retrovirus in birds, it was determined that lack of function of neuregulin 1 (NRG1) or their ErbB receptors not only causes deficient differentiation of ventricular myocytes and poor formation of trabeculae, but also determines premature death of the embryos. Based on this background, the aim of this work was to determine the actual role of NRG1 in early trabeculogenesis and its importance in proliferation and apoptosis regulation. Methods. An embryonic chicken heart organ culture system at the age prior to the beginning of the trabeculogenesis process was established. Endogenous activity of NRG1 was inhibited in the organ cultures that were then stimulated with NRG1 at different concentrations. Myocyte proliferation was determined using the proliferating cell nuclear antigen and apoptosis with LysoTracker (LTR). Results. Fetal bovine serum promotes proliferation but negatively impacts trabeculogenesis. Low concentration of NRG1 and short periods of incubation do not induce trabeculogenesis. In contrast, average NRG1 concentrations and cultivation periods not exceeding 24 h have a positive effect on the onset of this process. This also promotes myocardial proliferation but avoids apoptosis. Higher concentrations of NRG1 possibly cause a molecular imbalance that favors untidy proliferation but not trabeculogenesis. Conclusions. Understanding of the role of NRG1 on ventricular trabeculogenesis provides valuable information for the molecular pathways also involved in IVS development. This information is essential for understanding the origin of serious ventricular septal defects.

Salmonella infects B cells by macropinocytosis and formation of spacious phagosomes but does not induce pyroptosis in favor of its survival.

We have previously reported that Salmonella infects B cells and survives within endosomal-lysosomal compartments. However, the mechanisms used by Salmonella to enter B cells remain unknown. In this study, we have shown that Salmonella induces its own entry by the induction of localized ruffling, macropinocytosis, and spacious phagosome formation. These events were associated with the rearrangement of actin and microtubule networks. The Salmonella pathogenesis island 1 (SPI-1) was necessary to invade B cells. In contrast to macrophages, B cells were highly resistant to cell death induced by Salmonella. These data demonstrate the ability of Salmonella to infect these non-professional phagocytic cells, where the bacterium can find an ideal intracellular niche to support persistence and the possible dissemination of infection.

Identification of Fructose-1,6-bisphosphate aldolase cytosolic class I as an NMH7 MADS domain associated protein.

We are interested in identifying proteins that interact with the MADS domain protein NMH7 of Medicago sativa. We use an affinity column with a synthetic peptide derived from the MADS domain of NMH7 which has been reported to mediate protein-protein interaction with non-MADS domain interacting proteins. We identified approximately 40 and approximately 80kDa specifically bound proteins as the monomeric and dimeric forms of Fructose-1,6-bisphosphate aldolase cytosolic class I. NiNTA pull down assays revealed that K- and C-terminus regions of NMH7 are not required for the interaction with aldolase. Aldolase enzymatic activity is not required for the interaction with NMH7. NMH7 and aldolase were coimmunoprecipitated from non-inoculated seed and seedlings extracts. Colocalization studies using confocal microscopy showed that aldolase and NMH7 are localized in the cytoplasm and the nucleus of the cortical cells. These data together show that M. sativa aldolase is a novel MADS domain binding protein, and suggest a broader functional repertory for this enzyme, as has been proposed for other glycolytic enzymes.

Los segmentos cardíacos primitivos, su implicación en la cardiogénesis normal aplicada a la cardiología pediátrica / Involvement of the primitive cardiac segments in the normal cardiogenesis. Its importance in the pediatric cardiology

The ample development of diagnostic echocardiography in pediatric cardiology has demanded precise knowledge of the abnormal anatomy of hearts that present congenital cardiac diseases. As a result, the information on morphologic and molecular aspects of cardiac embryogenesis has become fundamental to understand the anomalous anatomy of the malformed hearts. Based on these facts, in this paper we reviewed normal cardiogenesis, integrating the new information obtained experimentally in the chick embryo and from classic descriptive knowledge in humans. The age at which each cardiac segment appears is specified. At the same time, the changes in shape, relationships and position of these cardiac segments are detailed. Some implications of this process in the production of congenital cardiac defects and the importance of some specific genes are also discussed. This information is useful in the diagnosis of congenital cardiac diseases, as well as in discussing their embryogenesis. It is also beneficial in studying the possible mechanisms and genes implicated in normal morphogenesis of cardiac chambers, septa and valves. All this knowledge is important to plan strategies to avoid the production of this type of congenital pathologies.

[Involvement of the primitive cardiac segments in the normal cardiogenesis. Its importance in the pediatric cardiology]. / Los segmentos cardíacos primitivos, su implicación en la cardiogénesis normal aplicada a la cardiología pediátrica.

The ample development of diagnostic echocardiography in pediatric cardiology has demanded precise knowledge of the abnormal anatomy of hearts that present congenital cardiac diseases. As a result, the information on morphologic and molecular aspects of cardiac embryogenesis has become fundamental to understand the anomalous anatomy of the malformed hearts. Based on these facts, in this paper we reviewed normal cardiogenesis, integrating the new information obtained experimentally in the chick embryo and from classic descriptive knowledge in humans. The age at which each cardiac segment appears is specified. At the same time, the changes in shape, relationships and position of these cardiac segments are detailed. Some implications of this process in the production of congenital cardiac defects and the importance of some specific genes are also discussed. This information is useful in the diagnosis of congenital cardiac diseases, as well as in discussing their embryogenesis. It is also beneficial in studying the possible mechanisms and genes implicated in normal morphogenesis of cardiac chambers, septa and valves. All this knowledge is important to plan strategies to avoid the production of this type of congenital pathologies.

Histological study of the proximal and distal segments of the embryonic outflow tract and great arteries.

The normal development of the ventricular outlets and proximal region of the great arteries is a controversial subject. It is known that the conus, truncus arteriosus (truncus), and aortic sac participate; however, there are some doubts as to the actual prospective fate of the truncus. Some authors propose that it gives origin to the proximal region of the great arteries and that the myocardial cells of its wall become smooth muscle. Nevertheless, others think that the truncus only forms the arterial valve apparatus and that therefore the myocardial cells transform into fibroblasts. As a first approach to beginning to elucidate which process occurs, the aim of this article was to study the histological changes in the wall of these components of the developing heart in chick embryos whose hearts had been labeled at the truncoconal boundary at stage 22HH, tracing the changes up to stage 36HH. Also, the histological constitution of the wall of the pulmonary arterial trunk and its valve apparatus were studied in the posthatching and adult hearts of chickens and rats. The conus and truncus walls were always encircled by a myocardial sleeve from the outset of their development. Between stages 26HH to 28HH, the truncal myocardial cells adjacent to the mesenchymal tissue of the ridges began to lose cell-to-cell contacts and invaded the extracellular matrix. At stage 24HH, the aortic sac began to project into the pericardial cavity and became divided into two channels by the aortic-pulmonary septum at stage 26HH. The wall of the aortic sac is mostly constituted by a compact mesenchymal tissue. Initially, it does not have smooth muscle but this starts to appear at stage 30HH. The insertion ring of the valves, a broad structure, was formed by mesenchymal tissue. Both structures were always covered by a myocardial sleeve. The leaflets developed from the truncal ridges, the segment immediately proximal to the aortic sac. Our results indicate that the proximal region of the pulmonary and aortic arteries do not originate from the truncus arteriosus; rather, we found that they take origin from the aortic sac. Thus, our findings agree with the proposal that the myocardial cells of the external sleeve of the truncus become fibroblastic and suggest that the insertion ring of the arterial valves has a dual origin: fibroblasts produced by truncal myocardial transdiferentiation and the mesenchymal tissue of the proximal region of the truncal ridges, while the leaflets have their origin from the truncal ridges. We discuss the fact that, because the truncus arteriosus does not give origin to the trunks of the aortic and pulmonary arteries, it may be necessary to modify terminology. Based on our results, together with the new findings obtained by in vivo labeling, immunostaining, a chimeric approach, and ultrastructural studies, we propose a developmental model that correlates the fate of the conus, truncus, and aortic sac with the normal morphogenesis of the ventricular outlet tracts and the trunks of the great arteries. (c) 2005 Wiley-Liss, Inc.

Consideraciones embriológicas y anatómicas sobre la septación cardiaca normal y patológica: II. Septum atrioventricular e interventricular / Embriologycal and anatomycal conciderations on normal and pathological cardiac septation. II atrioventricular and interventricular septa (AU)

Se define el septum atrioventricular y sus características anatomicas. Este septum se divide en dos regiones: muscular y membranosa. Se describen los tipos de comunciación atrioventricular aisladas. Se discute la embriogénesis normal y patológica del septum atrioventricular. Se define el septum interventricular y se describen sus características anatómicas. Este septum se divide en dos regiones: fibrosa y muscular. La muscular se divide en tres regiones: porción de la cámara de salida o infundibular (outlet). Las comunicaciones inteventriculares se clasifican de acuerdo con las regiones septales en que están ubicadas. Se establecen las características anatómicas de identidad de cada una de ellas. Se hace un breve comentario sobre la morfogénesis normal y patológica del septum interventricular. Septum atrioventricular; septum interventricular