Perinatal outcome and placental apoptosis in patients with late-onset pre-eclampsia and abnormal uterine artery Doppler at diagnosis.

OBJECTIVE: To determine the rate of placental apoptosis and adverse perinatal outcome in patients with late-onset pre-eclampsia (PE) and abnormal uterine artery (UtA) Doppler at diagnosis. METHODS: This was a prospective cohort study of women with singleton pregnancy diagnosed with late PE, performed between August 2011 and January 2014 at the Maternal-Fetal Medicine Unit of Hospital Carlos Van Buren. Patients were stratified according to UtA Doppler status at diagnosis (pulsatility index (PI) ≤ or > 95th percentile). Logistic regression analysis was performed to identify associations between abnormal UtA Doppler and adverse maternal and perinatal outcomes. In a subset of this cohort for whom placental samples were available, immunohistochemical analysis of the placenta was performed to identify the rate of apoptosis and its association with UtA Doppler by comparing samples from those with normal and those with abnormal UtA Doppler and normotensive controls. Non-parametric linear trend analysis was performed for assessment of the apoptotic index. RESULTS: Eighty-six patients were included in the final analysis. Of these, UtA-PI was above the 95th percentile in 33 (38.4%) patients. Gestational age at diagnosis and delivery were significantly lower in this group compared with patients with normal UtA Doppler. Abnormal UtA Doppler was associated with increased risk of severe PE (odds ratio (OR) = 7.5; 95% CI, 2.76-20.46; P < 0.001), late preterm delivery (OR = 13.7; 95% CI, 4.53-41.46; P < 0.001), small-for-gestational age at birth (OR = 12.3; 95% CI, 3.17-47.57; P < 0.001) and admission to the neonatal intensive care unit (OR = 12.8; 95% CI, 2.61-62.36; P = 0.002). Moreover, UtA Z-score demonstrated a significant inverse correlation with birth-weight Z-score (r = -0.34; P = 0.0013). Mean placental apoptotic index demonstrated an ascending linear trend according to UtA Doppler status (P = 0.04). CONCLUSIONS: In patients with late PE, UtA Doppler was useful for clinical classification and as an indicator of placental histological findings. Correlation between UtA Doppler and the apoptotic index provides new evidence of a subgroup of late PE with a placental origin. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

The human lung during the embryonic period: vasculogenesis and primitive erythroblasts circulation.

Vascularization and blood cell circulation are crucial steps during lung development. However, how blood vessels are generated and when lung circulation is initiated is still a matter of debate. A morpho-functional analysis of pulmonary vasculature was done using human lung samples between 31 and 56 days post-fertilization (pf). The immunolocalization and expression of CD31, CD34, FLT-1, KDR and the vascular growth factor (VEGF) were investigated. The results showed that at day 31 pf, a capillary plexus is already installed, and a few primitive erythroblasts were seen for the first time within the lumen of some blood vessels. Around day 45 pf, an increase in the amount of primitive erythroblasts was detected in the parenchyma surrounding the distal segment of the bronchial tree. The expression of FLT-1, KDR, CD31 and CD34 was observed in endothelial cells of the capillary plexus and the VEGF was detected in the endodermal epithelium. Our results support the hypothesis that the initial formation of the capillary plexus around the tip of the growing airway bud occurs by vasculogenesis, probably regulated by VEGF and KDR. We also showed a very early onset of blood circulation, starting from day 34 pf, concomitant with the generation of new lung buds. In addition, the increasing number of primitive erythroblasts from week 6 onward, associated with a change in the shape of the blood vessels, suggests a remodeling process and that the generation of new distal vessels at the tip of the lung bud occurs mainly by a process of angiogenesis.

Effect of normobaric hypoxia on the testis in a murine model.

High-altitude hypoxia generates spermiogram impairment due to germinal epithelium, Leydig cells, sperm and seminal plasma alterations, but precise mechanisms involved are unknown. The objective of this work was to analyse the effect of normobaric hypoxia on the morphology of testicular interstitium and some associated molecular and hormonal factors. Twenty-four mice were exposed to normobaric hypoxia (8.1% inspired oxygen fraction) during 20 days. The effects on body weight, testicular weight, vascularisation, testosterone, HIF1-α and VEGF were analysed at different periods of exposure and compared to controls. Hypoxic mice had lower body weight than mice kept in normoxia. Testicular weight raised significantly the 1st day, but remained normal during the rest of experiment. Number of blood vessels per field and mean diameter of vessels were higher in hypoxic mice. Plasmatic and testicular testosterone raised during first 24 h of hypoxia, but decreased on the 5th day. Vascular/interstitial ratio (proportion of interstice occupied by blood vessels) duplicated at the end of the experiment. Most substantial early effects of hypoxia were testicular oedema, increase in number and diameter of blood vessels and elevation of plasmatic and testicular testosterone. Normobaric hypoxia generates similar effects to those induced by hypobaric hypoxia.

Prognostic value of plasma pentraxin 3 levels in patients with septic shock admitted to intensive care.

OBJECTIVE: To evaluate the usefulness of a new marker, pentraxin, as a prognostic marker in septic shock patients. MATERIALS AND METHODS: Single-centre prospective observational study that included all consecutive patients 18 years or older who were admitted to the intensive care unit (ICU) with septic shock. Serum levels of procalcitonin (PCT), C-reactive protein (CRP) and pentraxin (PTX3) were measured on ICU admission. RESULTS: Seventy-five septic shock patients were included in the study. The best predictors of in-hospital mortality were the severity scores: SAPS II (AUC = 0.81), SOFA (AUC = 0.79) and APACHE II (AUC = 0.73). The ROC curve for PTX3 (ng/mL) yielded an AUC of 0.70, higher than the AUC for PCT (0.43) and CRP (0.48), but lower than lactate (0.79). Adding PTX3 to the logistic model increased the predictive capacity in relation to SAPS II, SOFA and APACHE II for in-hospital mortality (AUC 0.814, 0.795, and 0.741, respectively). In crude regression models, significant associations were found between in-hospital mortality and PTX3. This positive association increased after adjusting for age, sex and immunosuppression: adjusted OR T3 for PTX3 = 7.83, 95% CI 1.35-45.49, linear P trend = 0.024. CONCLUSION: Our results support the prognostic value of a single determination of plasma PTX3 as a predictor of hospital mortality in septic shock patients.

[Multicenter study on the monitoring of in vitro susceptibility to tigeeyeline in Santiago, Chile]. / Estudio multicéntrico de la vigilancia de la susceptibilidad in vitro a tigeciclina en Santiago de Chile.

The objective of this multicenter study was to determine tigecycline susceptibility rates, measured by agar diffusion, in nine hospitals in Santiago and to compare these rates with other antimicrobials. Each center studied 20 strains per month. All intermediate and fully resistant strains as well as 10% of susceptibile strains were also studied by the broth microdilution method. Overall, 2301 strains were studied displaying the following susceptibility rates for tigecycline: 100% for Streptococcus sp, Enterococcus sp, and E. coli respectively, 99.8% for Staphylococcus sp, 93% for Klebsiella and 80% for Acinetobacter baumarmii. For Proteus, Providencia and Morganella the susceptibility rates were 4%. For cefotaxime-resistant Klebsiella and imipenem-resistant A. baumarmii susceptibility rates were 95% and 80% respectively. The agar diffusion and broth dilution method were 100% concordant for tigecycline susceptible strains but only 27% for resistant or intermediate strains represented mostly by Acinetobacter baumannii. The majority of these strains (57/59) proved to be susceptible after retesting. The great majority (96,6%) of strains tested from nine Chilean hospitals proved to be susceptible to tigecycline with exception for Proteus, Providencia and Morganella (66% resistance). Using the agar diffusion method for measuring tigecycline susceptibility to A. baumannii may be misleading.

Maternal diabetes affects specific extracellular matrix components during placentation.

During embryo implantation, invasive trophoblast cells mediate embryo invasion into the decidualized stroma, forming a rich network of lacunae that connect the embryonic tissues to the maternal blood vessels. Placentation is probably guided by the composition and organization of the endometrial extracellular matrix. Certain pathological conditions that occur during pregnancy, including diabetes, have been linked to abnormal placental morphology and consequent fetal morbidity. We used immunoperoxidase techniques to identify members of the collagen, proteoglycan and glycoprotein families in the various compartments of the rat placenta and to determine whether experimentally induced diabetes affects placental morphology and alters the distribution of these molecules during pregnancy. Single injections of alloxan (40 mg kg(-1) i.v.) were used to induce diabetes on day 2 of pregnancy in Wistar rats. Placentas were collected on days 14, 17, and 20. Type I and III collagen, as well as the proteoglycans decorin and biglycan, were found to be distributed throughout the placentas of control and diabetic rats. In both groups, laminin expression decreased at the end of pregnancy. In contrast, fibronectin was detected in the labyrinth region of diabetic rats at all gestational stages studied, whereas it was detected only at term pregnancy in the placentas of control rats. These results show for the first time that some extracellular matrix molecules are modulated during placental development. However, as diabetic rats presented increased fibronectin deposition exclusively in the labyrinth region, we speculate that diabetes alters the microenvironment at the maternal-fetal interface, leading to developmental abnormalities in the offspring.

Transplantation of Human Amniotic Membrane over the Liver Surface Reduces Hepatic Fibrosis in a Cholestatic Model in Young Rats.

PURPOSE: Biliary atresia precedes liver cirrhosis and liver transplantation. Amniotic membrane (AM) promotes tissue regeneration, inhibits fibrosis, and reduces inflammation. Here, we test amniotic membrane potential as a therapeutic tool against cholestatic liver fibrosis. METHODS: Three groups of rats were used: sham surgery (SS), bile duct ligature (BDL), and bile duct ligature plus human amniotic membrane (BDL + AM). After surgery, animals were sacrificed at different weeks. Biochemical and histopathological analyses of liver tissue were performed. Collagen was expressed as a percentage of total liver tissue area. qPCR was performed to analyse gene expression levels of transforming growth factor-ß1 (Tgfb1) and apelin (Apln). Statistical analysis performed considered p < 0.05 was significant. RESULTS: Groups undergoing BDL developed cholestasis. Biochemical markers from BDL + AM group improved compared to BDL group. Ductular reaction, portal fibrosis, and bile plugs were markedly reduced in the BDL + AM group compared to BDL group. Collagen area in BDL + AM group was statistically decreased compared to BDL group. Finally, expression levels of both Apln and Tgfb1 mRNA were statistically downregulated in BDL + AM group versus BDL group. CONCLUSION: AM significantly reduces liver fibrosis in a surgical animal model of cholestasis. Our results suggest that AM may be useful as a therapeutic tool in liver cirrhosis.

Presence of tumor DNA in plasma of breast cancer patients: clinicopathological correlations.

Using different molecular techniques, DNA has been detected in the plasma of cancer patients with various types of tumors. We undertook the present study to investigate the presence of plasma DNA, before mastectomy, in patients with breast cancer at diagnosis and to analyze the clinicopathological spectrum of this subgroup of patients with respect to patients without DNA with tumor characteristics. We studied 62 patients with breast cancer, who were selected sequentially after mastectomy and diagnosis of breast carcinomas. Genomic DNA extracted from tumor and normal tissues, normal blood cells, and plasma was used for molecular studies. Alterations in polymorphic markers selected because they had been found to show a high rate of alterations in breast cancer in previous studies (D17S855, D17S654, D16S421, TH2, D10S197, and D9S161), as well as mutations in the p53 gene and aberrant methylation at the first exon of p16INK4a, were used to identify and characterize tumor and plasma DNA. Thirteen clinicopathological parameters were analyzed in each patient. We identified 56 cases (90%) with at least one molecular event in tumor DNA, and 41 cases (66%) with a similar alteration in plasma DNA. Comparison of the clinicopathological parameters between patients with and without plasma DNA revealed significant differences in the axillary involvement, rate of invasive ductal carcinoma, high proliferative index, and the parameter comprised of lymph node metastases, histological grade II, and peritumoral vessel involvement. A high proportion of breast cancer patients exhibited plasma DNA at diagnosis similar to tumor DNA, and its presence correlated significantly with pathological parameters associated with a poor prognosis.

[Fusarium species of the Quebrada de Lozano, Jujuy, Argentina]. / Especies de Fusarium de la Quebrada de Lozano, Jujuy, Argentina.

A soil sampling was made in 50 cultivated places with horticultural and floral species where there were fusariosis records. For the isolation Bengal rose agar, peptone dichloran chloramphenicol agar and cayote dichloran chloramphenicol agar were used, being this last one, the more convenient. The isolates were identified as F. acuminatum, F. avenaceum, F. graminearum, F. merismoides, F. oxysporum, F. poae, F. proliferatum, F. semitectum, F. solani, F. subglutinans and F. sporotrichioides. It was proven the pathogenesis of F. acuminatum and F. proliferatum strains isolated from cultivated places with Cucurbita ficifolia and Vicia faba respectively, and of the strains of F. oxysporum obtained of soils with cultivations of Chrysanthemum leucanthemum, Ocimum basilicum and Zea mays. The F. merismoides, F. semitectum and F. subglutinans strains coming from places with these three vegetable species, were recovered as secondary invaders.

A study of the electric field distribution in erythrocyte and rod shape cells from direct RF exposure.

This paper shows the importance of using realistic cell shapes with the proper geometry and orientation to study the mechanisms of direct cellular effects from radiofrequency (RF) exposure. For this purpose, the electric field distribution within erythrocyte, rod and ellipsoidal cell models is calculated by using a finite element technique with adaptive meshing. The three cell models are exposed to linearly polarized electromagnetic plane waves of frequencies 900 and 2450 MHz. The results show that the amplification of the electric field within the membrane of the erythrocyte shape cell is more significant than that observed in other cell geometries. The results obtained show the dependence of the induced electric field distribution on frequency, electrical properties of membrane and cytoplasm and the orientation of the cell with respect to the applied field. The analysis of the transition of an erythrocyte shape to an ellipsoidal one shows that a uniformly shelled ellipsoid model is a rough approximation if a precise simulation of bioeffects in cells is desired.

Modelling the internal field distribution in human erythrocytes exposed to MW radiation.

This paper studies the internal electric field distribution in human erythrocytes exposed to MW radiation. For this purpose, an erythrocyte cell model is exposed to linearly polarized electromagnetic (EM) plane waves of frequency 900 MHz and the electric field within the cell is calculated by using a finite element (FE) technique with adaptive meshing. The results obtained show the dependence of the induced electric field distribution on the main modelling parameters, i.e., the electrical properties (permittivity and conductivity) of the membrane and cytoplasm and the orientation of the cell with respect to the applied field. It is found that for certain orientations, the field amplification within the membrane of the erythrocyte shape cell can be higher than the one observed in an equivalent simple spheroidal geometry cell, commonly used in bioelectromagnetism. The present work shows that a better insight of the interaction of electromagnetic fields with basic biological structures is obtained when the most possible realistic cell shape is used.

Distribution of versican and hyaluronan in the mouse uterus during decidualization.

Preparation for embryo implantation requires extensive adaptation of the uterine microenvironment. This process consists of cell proliferation and cell differentiation resulting in the transformation of endometrial fibroblasts into a new type of cell called decidual cell. In the present study, we followed the space-time distribution of versican and hyaluronan (HA) in different tissues of the uterus before and after embryo implantation. Fragments of mouse uteri obtained on the fourth, fifth, sixth and seventh days of pregnancy were fixed in Methacarn, embedded in Paraplast and cut into 5-microm thick sections. HA was detected using a biotinylated fragment of the proteoglycan aggrecan, which binds to this glycosaminoglycan with high affinity and specificity. Versican was detected by a polyclonal antibody. Both reactions were developed by peroxidase methods. Before embryo implantation, both HA and versican were present in the endometrial stroma. However, after embryo implantation, HA disappeared from the decidual region immediately surrounding the implantation chamber, whereas versican accumulated in the same region. The differences observed in the expression of HA and versican suggest that both molecules may participate in the process of endometrial decidualization and/or embryo implantation.

[The most significative parameters for the diagnosis of focal neuropathy of the cubital nerve in the elbow]. / Parámetros más significativos para el diagnóstico de la neuropatía focal del nervio cubital en el codo.

OBJECTIVE: Determine the diagnostic validity of the most usual neurophysiological procedures in the examination of compressive neuropathies and/or ulnar nerve entrapment at the elbow. PATIENTS AND METHODS: A prospective observational study was carried out over a period of one year which included 19 patients with clinical features compatible with entrapment/compression of the ulnar nerve at the elbow, their neurophysiological diagnosis and surgical confirmation. We also studied 49 patients who were attended in our clinic for other reasons not related to disorders of the ulnar nerve. The variables studied were: maximum rate of motor conduction above the elbow; the amplitudes between spikes and the area between the first negative deflection and the last positive spike of the responses at the wrist and above the elbow. RESULTS: In the group of persons without disorders of the ulnar nerve the average rate of motor conduction was 58.07 m/s, with a confidence interval of 53-65. In the group of patients with specific clinical features the average rate of motor conduction was 47.26 m/s, with a confidence interval of 43.74-50.77. No relation was found between age and pathology, although rate of motor conduction was related to the presence of pathology. CONCLUSIONS: It was seen on logistical regression that the main variables for prediction of the presence or absence of pathology in a person were firstly motor conduction and secondly the amplitude index.

[Stage II non-seminoma cancer of the testis: combined treatment with surgery and chemotherapy]. / Cáncer testicular no seminoma etapa II: tratamiento combinado con cirugía y quimioterapia.

Fifty-one patients with non-seminoma testicular cancer who underwent retroperitoneal ganglion dissection and 3 support cycles of chemotherapy with Cys-Platin, Vinblastine and Bleomycin. Four patients presented relapses (7.8%). Actuarial survival by 8 years was 87%, with a median of 38 months. The median survival was 43 months. Three of the 4 patients with relapses are currently free of disease after rescue treatment, showing in global terms a control of the disease of 98% during this clinical stage.

Lección MOODLE de anatomía e histología de la placenta humana / MOODLE lesson of the anatomy and histology of the human placenta

RESUMEN: La falta de muestras biológicas humanas existentes, debido principalmente a las limitaciones ético-morales relacionadas con su obtención, ponen en relieve la necesidad de buscar otras alternativas de enseñanza y aprendizaje de las ciencias morfológicas. En este sentido, la implementación de lecciones a través de la plataforma MOODLE proporciona la oportunidad al estudiante de interactuar en un entorno que simula una situación de aprendizaje propio del laboratorio tradicional. El objetivo del presente trabajo fue generar una lección MOODLE sobre la anatomía e histología placentaria humana, como complemento a la clase teórica presencial, para estudiantes de la carrera de Obstetricia y Puericultura. Para tal cometido, se realizó búsqueda de información, imágenes y recursos TIC en bibliotecas e internet. Paralelamente, se llevó a cabo un proceso de captura fotográfica de muestras histológicas de placenta, así como también la grabación de un alumbramiento. Posteriormente, se procedió a la articulación y montaje de las actividades en la plataforma MOODLE con un enfoque constructivista. Además, se elaboró una encuesta de satisfacción, la cual fue validada por 3 expertos. La muestra estuvo constituida por 137 estudiantes de la carrera de Obstetricia. Se confeccionó un laboratorio virtual MOODLE de anatomía e histología de la placenta humana, el cual esta constituido por múltiples actividades con orientación clínica, las cuales permiten autoevaluarse. El laboratorio virtual nos ha ayudado ha subsanar la carencia de muestras humanas y los resultados de la encuesta de satisfacción aplicada a los estudiantes señalan una valoración positiva de la iniciativa.

SUMMARY: The lack of existing human biological samples, mainly due to the ethical-moral restrictions related to obtaining these, highlights the need to search for other teaching and learning alternatives in morphological science. In this sense, the implementation of lessons by means of the MOODLE platform provides the students with the opportunity to interact in a setting that simulates a learning situation that belongs to traditional laboratories. The purpose of this work was to generate a MOODLE lesson on the anatomy and histology of the human placenta, as a complement of the traditional theoretical classroom for students of Obstetrics. To that end, TIC information, images, and resources were sought in libraries and in the Internet, and at the same time a set of histological photographs of placenta samples was made, as well as a video recording of a placental delivery. Later, the coordination and set up of activities was made in the MOODLE platform with a constructivist approach. Furthermore, a satisfaction survey was prepared which was validated by three experts. The total sample consisted of 137 students in the 2th year of obstetrics. A virtual MOODLE laboratory of the anatomy and histology of the human placenta was made, which is constituted by multiple activities with a clinical orientation that allow self-evaluation. The virtual laboratory has helped overcome the lack of human samples, and results of the satisfaction survey applied to the students indicate a positive evaluation of this initiative.

Increased placental angiogenesis in late and early onset pre-eclampsia is associated with differential activation of vascular endothelial growth factor receptor 2.

INTRODUCTION: Placentas from both early-onset (EOPE) and late-onset pre-eclampsia (LOPE) exhibit signs of underperfusion, which in turn, may be associated with altered angiogenesis. Tyrosine 951 (Y951) and Y1175 phosphorylation of the vascular endothelial growth factor receptor 2 (VEGFR2) induced by VEGF triggers the angiogenesis process. Endothelial markers such as CD31 and CD34 have been used for estimating angiogenic processes in several tissues, including placenta. We asked whether vascular density in placental villi was related to Y951/Y1175 phosphorylation of VEGFR2 in LOPE or EOPE. METHODS: We obtained placental samples from women with normal pregnancies (n = 22), LOPE (n = 13), EOPE (n = 15) and preterm deliveries (n = 10). Slices from placental tissue were used for CD31 immunostaining. We estimated the expression of CD31, CD34, VEGF, and VEGFR2 by western blot and quantitative PCR. Y951 phosphorylation of VEGFR2 was estimated by western blot, whereas Y1175 phosphorylation was analyzed by ELISA. RESULTS: Vessel density in terminal villi and CD31 and CD34 protein abundance were increased in LOPE and EOPE compared to normal pregnancy. However, mRNA levels for CD31 and CD34 were lower in LOPE than in normal pregnancy and VEGF mRNA was higher in EOPE. VEGFR2 protein concentration was not different among the studied groups. Y951 and Y1175 phosphorylation of VEGFR2 was higher in LOPE than in the normotensive group, but only Y951 exhibited greater phosphorylation in EOPE compared to normal pregnancy. DISCUSSION: Changes in vessel formation in the pre-eclamptic placenta are controversial. Our study suggests a pro-angiogenic state in both LOPE and EOPE. These changes are however, associated with differential expression of endothelial markers and VEGFR2 activation. CONCLUSION: There is evidence of increased placental angiogenesis in LOPE and EOPE that is associated with differential activation of VEGFR2.

Stat3 and Socs3 expression patterns during murine placenta development.

Signal transducers and activators of transcription 3 (Stat3) has been identified as an important signal transducer in the invasive phenotype of the trophoblasts cells in in vitro studies. However, the in situ distribution and patterns of expression of this molecule in trophoblast cells during the development of the placenta are still under-elucidated. Mice uteri of gestational ages between 7 and 14 days of pregnancy (dop) were fixed in methacarn and processed with immunoperoxidase techniques for detection of Stat3 and its phosphorylation at serine (p-ser727) residues, as well as the suppressor of cytokine signaling 3 (Socs3) expression. Stat3 was observed at 7 through 9 dop in both the antimesometrial and mesometrial deciduas, while continued immunoreactivity between 10 and 13 dop was seen only in the mesometrial decidua. In the placenta, Stat3 was detected in the cytotrophoblast cells of labyrinth and giant trophoblast cells between 10 and 14 dop. Immunoreactivity for Stat3 was also seen in trophoblast cells surrounding the maternal blood vessels. On days 10 and 11 of pregnancy, p-ser727 was detectable in the mesometrial decidua and in giant trophoblasts, while during 12-14 dop in the spongiotrophoblast region. In addition, Socs3 was immunodetected in maternal and placental tissues, principally in the giant trophoblast cells during the whole period of the study. The present in situ study shows the distribution of Stat3, its serine activation and Socs3 in different maternal and fetal compartments during murine placental development, thus further supporting the idea that they play a role during physiological placentation in mice. 

The effects of fibrin and fibrin-agarose on the extracellular matrix profile of bioengineered oral mucosa.

Several studies have developed efficient oral mucosa constructs using different types of scaffold. However, the changes in the morphology and gene and protein expression profile that could occur in these artificial constructs remain unknown. This study compared the histology and expression of several extracellular matrix molecules in human artificial oral mucosa developed using two different types of scaffolds: fibrin and fibrin-agarose. To that end, bioengineered oral mucosa stromas were constructed from biopsy samples of human oral mucosa and the substitute generated was analyzed at different periods of time in culture. Histological analysis was carried out by light and transmission electron microscopy and the expression of collagen types I, III, and VI, the proteoglycans decorin and biglycan, and the different chains of laminin, were assessed by immunoperoxidase technique. This study found that fibrin scaffolds accelerated fibroblast growth and remodeling of the scaffold, thus enhancing collagen fibrillogenesis. In the fibrin-agarose scaffold, the morphology and organization of the fibroblasts did not change during the culture period. All extracellular matrix proteins analyzed were expressed in both scaffolds. However, in fibrin scaffolds, these proteins were widely distributed and replaced the scaffold during the follow-up period. These results show that the substitutes generated showed histological and molecular similarities with native human oral mucosa stroma. In addition, it was observed that the nature of the biomaterial influenced the behaviour of the oral stromal fibroblasts, thereby modulating their growth, protein synthesis, and collagen fibrillogenesis.

Proteoglycan and collagen expression during human air conducting system development.

The lung is formed from a bud that grows and divides in a dichotomous way. A bud is a new growth center which is determined by epithelial-mesenchymal interactions where proteins of the extracellular matrix (ECM) might be involved. To understand this protein participation during human lung development, we examined the expression and distribution of proteoglycans in relation to the different types of collagens during the period in which the air conducting system is installed. Using light microscopy and immunohistochemistry we evaluate the expression of collagens (I, III and VI) and proteoglycans (decorin, biglycan and lumican) between 8 to 10 weeks post fertilization and 11 to 14 weeks of gestational age of human embryo lungs. We show that decorin, lumican and all the collagen types investigated were expressed at the epithelium-mesenchymal interface, forming a sleeve around the bronchiolar ducts. In addition, biglycan was expressed in both the endothelial cells and the smooth muscle of the blood vessels. Thus, the similar distribution pattern of collagen and proteoglycans in the early developmental stages of the human lung may be closely related to the process of dichotomous division of the bronchial tree. This study provides a new insight concerning the participation of collagens and proteoglycans in the epithelial-mesenchymal interface during the period in which the air conducting system is installed in the human fetal lung.