Histology and histopathology Vol.34, nº3 (2019)
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- PublicationOpen AccessDouble immunofluorescence labeling for CD31 and CD105 as a marker for polyether polyurethane-induced angiogenesis in mice(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Figueiredo, Camila Couto; Braga Pereira, Núbia; Pereira, Luciana Xavier; Machado Oliveira, Laser Antônio; Peixoto Campos, Paula; Passos Andrade, Silvia; Moro, LucianaA crucial component of the integration between foreign implants and the host is angiogenesis. However, to date, none of the available techniques and/or endothelial markers employed to assess angiogenesis in the implant/host interface seems to be able to highlight vascular structures convincingly. In the present study we investigated and compared the expression of two endothelial cell markers: platelet endothelial cell adhesion molecule (PECAM-1) (CD31) and endoglin (CD105) using immunohistochemistry (IHC) and immunofluorescence (IF) to identify and quantify newly formed blood vessels in subcutaneous implants of polyether-polyurethane sponge of formalin-fixed paraffin-embedded tissue. At day 14 post implantation the discs of the synthetic matrix were removed and processed for histological and morphometric analysis. In IHC staining for CD31 antibody the number of vessels was 2.27±0.69 and 5.25±0.46 for CD105. In IF for CD31 the number of vessels was 15.36±1.295 and 10.54±0.8213 for CD105. The level of cross-reaction was lesser in IF images compared with IHC images. Colocalization of CD31/CD105 using confocal images showed positive correlation (Pearson's co-relation and Manders’ equation). The double labeling for blood vessels using the IF technique for CD31/CD105 may be an important tool for evaluation of angiogenesis in biomaterial/host integration.
- PublicationOpen AccessMMP-1/TIMP-1 expressions in rectal submucosa of females with obstructed defecation syndrome associated with internal rectal prolapse(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Cao, Long-Lei; Yu, Jie; Yang, Zhang-Ling; Qiao, Xu; Ye, Hui; Xi, Chang-Lei; Zhou, Qi-Chang; Hu, Cheng-Cheng; Zhao, Chao-Jun; Gong, Zhi-LinObjective. To explore the MMP-1/TIMP-1 expressions rectal submucosa of females with obstructed defecation syndrome (ODS) associated with internal rectal prolapse (IRP). Methods. Fifty-six female patients with ODS associated with IRP were enrolled as Case group, and 43 female hemorrhoids of stages III-IV without constipation and IRP were enrolled as Control group. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry were performed to test the expressions of MMP1/TIMP-1 in the rectal submucosa. Western blotting was used to examine protein expressions of MMP-1/TIMP-1 and pro-inflammatory cytokines (IL-6 and TNF-α) in the rectal submucosa. EVG staining was conducted to detect collagen and elastic fibers in rectal submucosa. Results. The increased expression of MMP-1 was negatively linked to the decreased TIMP-1 level in the rectal submucosa of patients with ODS associated with IRP. Besides, the expressions of IL-6 and TNF-α were increased in the Case group as compared with the Control group. Additionally, ODS severity and the proinflammatory cytokines was positively linked to MMP1, but negatively related to TIMP-1 in Case group. EVG staining showed that the area ratios of collagen and elastic fibers were lower in Case group than Control group. Through Pearson’s correlation analysis, the area ratios of collagen and elastic fibers were positively associated with MMP-1 expression, but negatively correlated with TIMP-1 expression in rectal submucosa of patients with ODS associated with IRP. Conclusion. Elevated MMP-1 and reduced TIMP-1 were found in ODS associated with IRP, which was related to the ODS severity, inflammation and contents of collagen and elastic fibers.
- PublicationOpen AccessComparison of histological characteristics and expression of CD3 and CD79a among the hemal nodes, lymph nodes and spleens of yaks (Bos grunniens)(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Huang, Yufeng; Cui, Yan; Yu, Sijiu; He, Junfeng; He, Yanyu; Zhang, Qian; Liu, Penggang; Pu, Yanhuan; Sun, Juan; Kang, XinhuaThis experiment compared the histological characteristics and distribution of CD3 and CD79a among yak lymph nodes, hemal nodes and spleen. The three organs from ten clinically healthy newborn and adult yaks were studied using histology and immunohistochemistry. The yak hemal nodes, which consisted of blood sinuses, lymphoid follicles, diffuse lymphoid tissue and lymphoid cords, appeared to share the histological characteristics of the spleen and lymph nodes: the lymphoid follicles of the hemal nodes were much like those of the lymph nodes, which were not surrounded by the central artery and periarteriolar lymphoid sheath. The lymphoid cords of the hemal node, which contained many erythrocytes, were much like the splenic cords. The sinuses of the hemal nodes had a similar structure to the lymph sinuses of the lymph nodes but were engorged with erythrocytes rather than lymph as in the lymph nodes. Interestingly, the splenic sinuses of yak were of two different types: the sinuses with obvious endothelial cells or those consisting of reticular cells. The CD3+ cells were mainly located in the paracortex area and medulla of the lymph nodes, the diffuse lymphoid tissues of the hemal nodes, and the periarteriolar lymphoid sheaths and red pulp of the spleen. Most CD79a+ cells were mainly detected in the lymphoid follicles of all examined lymphoid organs. The results suggested that although the three organs had specific characteristics, in some respects, they had similar organizational structural characteristics and immune functions. These may be useful to better understand the relationship between the morphology and function of these organs and provide useful references for normal yak lymphoid organs.
- PublicationOpen AccessThe effect of chronic stress and obesity on sperm quality and testis histology in male rats; a morphometric and immunohistochemical study(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Demirci, Tuba; Sahin, ElvanObjective. Stress and obesity lead structural and functional abnormalities in many organs. This research investigated the effect of these two risk factors on sperm parameters and histologic structure of testis in rat model. Materials and methods. Twenty-four male rats were divided into four groups with six rats in each group as control, stress, obesity, and stress-obesity groups. The rats of obesity and stress-obesity groups were fed highfat diet for 12-week and obesity was created. Control and stress groups were given standard rat chow for the same time. Through last 4-week, stress and stressobesity groups were applied to chronic mild stress procedure. At the end of the experiment, epididymal sperm was collected from vas deferens and testes were harvested from sacrificed animals. Sperm samples were evaluated in terms of concentration and motility by using Makler Chamber. Sperm smears were stained with Eosin-Y stain for morphological evaluation, and also histochemically for GABA transporter-1 (GAT1) expression assessment. Testis sections were dyed with Hematoxylin-Eosin and Johnsen scores were assessed. GAT1 expression was detected in testis sections by immunohistochemistry, and TUNEL method was used for determining apoptosis in testis. Results. In comparison with the control samples in stress, obesity, and stress-obesity groups sperm concentration and motility decreased, and also the number of sperm with abnormal morphology increased. Stress, obesity, and stress-obesity groups showed a significantly decreased in sperm concentration and motility in comparison with the control group, and also in these groups had significantly increased number of abnormal sperm compared to control. Additionally, the testicular structure was deteriorated, and Johnsen scores decreased. And also GAT1 expression and apoptosis were prominent. These negative results, especially, testicular weight, sperm concentration, and Johnsen score were more observed in the stress-obesity group. Conclusions. Stress and obesity may induce male infertility by disrupting both sperm quality and testis histology. When stress and obesity are coexisting, these adverse effects are more severe. And also, increased GAT1 expression may be associated with these effects.
- PublicationOpen AccessMultichromatic TTF staining characterizes cartilage matrix in osteoarthritis and bone development(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Feng, Yu; Cai, Zhe; Cheung, Wing ki; Yang, Kedi; Xu, Lei; Weijia Lu, William; Yang, Haibo; Chiu, Kwong-YuenVarious histological staining methods have been explored to detect the joint lesions in osteoarthritis (OA), but these histological stains cannot comprehensively present the comparatively complex structures of articular cartilage in knee OA. In addition, no integrated histological staining method can be used to evaluate efficiently both the subzone region and matrix composition in cartilage containing tissues. Therefore, in this study, a novel multichromatic staining method termed TTF staining, using Toluidine Blue (T), Tartrazine (T) and Fast Green (F) sequential combined staining for histological analysis, has been exploited to characterize the changes of matrix components and contents in cartilage during OA and in the bone development. This specific TTF staining profile can be used to differentiate the major compartments of knee joint region, including the synovium, meniscus, multiple subzones of cartilage and subchondral bone. An anterior cruciate ligament transection induced OA model in rat has been established to profoundly present the alterations of glycosaminoglycans in cartilage degeneration by TTF staining profile. The changes of TTF staining profile in the chondrification and ossification centers of the postnatal rat knee joint indicate the developmental features of cartilage matrix during the growth of bone. In summary, we have developed an effective histological staining method that enables us to identify the subzones of cartilage in detail and to define the matrix features of bone development. Therefore, finally using this new TTF staining method may help us to exploit a histopathological grading system to assess cartilage lesions in clinical disease.