Histology and histopathology Vol.31, nº9 (2016)
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- PublicationOpen AccessDamage to pancreatic acinar cells and preservation of islets of Langerhans in a rat model of acute pancreatitis induced by Karwinskia humboldtiana (buckthorn)(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Carcano Diaz, Katya; Garcia Garcia, Aracely; Segoviano Ramirez, Juan Carlos; Rodriguez Rocha, Humberto; Loera Arias, Maria de Jesus; Garcia Juarez, JaimeKarwinskia humboldtiana (Kh) is a poisonous plant that grows in some regions of the American continent. Consuming large amounts of Kh fruit results in acute intoxication leading to respiratory failure, culminating in death within days. There is evidence of histological damage to the lungs, liver, and kidneys following accidental and experimental Kh intoxication. To date, the microscopic effect of Kh consumption on the pancreas has not been described. We examined the early effects of Kh fruit on pancreatic tissue at different stages of acute intoxication in the Wistar rat. We found progressive damage confined to the exocrine pancreas, starting with a reduction in the number of zymogen granules, loss of acinar architecture, the presence of autophagy-like vesicles, apoptosis and inflammatory infiltrate. The pancreatic pathology culminated in damaged acini characterized by necrosis and edema, with a complete loss of lobular architecture. Interestingly, the morphology of the islets of Langerhans was conserved throughout our evaluations. Taken together, our results indicate the damage induced by a high dose of Kh fruit in the Wistar rat is consistent with an early acute necrotizing pancreatitis that exclusively affects the exocrine pancreas. Therefore, this system might be useful as an animal model to study the treatment of pancreatic diseases. More importantly, as the islets of Langerhans were preserved, the active compounds of Kh fruit could be utilized for the treatment of acinar pancreatic cancer. Further studies might provide insight into the severity of acute Kh intoxication in humans and influence the design of treatments for pancreatic diseases and acinar pancreatic cancer.
- PublicationOpen AccessTransfer of mesenchymal stem cells and cyclosporine A on alkali-injured rabbit cornea using nanofiber scaffolds strongly reduces corneal neovascularization and scar formation(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Cejka, Čestmír; Cejkova, Jitka; Trosan, Peter; Zajicova, Alena; Sykova, Eva; Holan, VladimirThe aim of this study was to examine whether nanofiber scaffolds seeded with rabbit bone marrow mesenchymal stem cells (MSCs nanofibers) transferred onto the damaged corneal surface and covered with cyclosporine A (CsA)-loaded nanofiber scaffolds (CsA nanofibers) enable healing of the rabbit cornea injured with 1N NaOH. The healing of damaged corneas was examined morphologically, immunohistochemically and biochemically on day 24 after the injury. Compared to untreated injured corneas, where corneal ulceration or large corneal thinning or even perforation were developed, injured corneas treated with drug free nanofibers healed without profound disturbances in a majority of cases, although with fibrosis and scar formation. In injured corneas treated with CsA nanofibers or MSCs nanofibers, the development of scar formation was reduced. Best healing results were obtained with a combination of MSCs and CsA nanofibers (MSCs-CsA nanofibers). Corneas healed with highly restored transparency. Neovascularization highly expressed in untreated injured corneas and reduced in corneas treated with CsA nanofibers or MSCs nanofibers, was suppressed in corneas treated with MSCs-CsA nanofibers. The levels of matrix metalloproteinase 9, inducible nitric oxide synthase, interleukin 6, α-smooth muscle actin, tumor growth factor β and vascular endothelial growth factor were significantly decreased in these corneas as compared to untreated corneas, where the levels of the above mentioned markers were high. In conclusion, MSCsCsA nanofibers were effective in the treatment of severe alkali-induced corneal injury.
- PublicationOpen AccessBlind aspiration biopsy versus a guided hysteroscopic technique for investigation of the endometrium in infertile women(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Ejzenberg, Dani; de Jesus Simões, Manuel; Pinheiro, Walter; Soares Júnior, José Maria; Serafini, Paulo Cesar; Chada Baracat, EdmundEmbryo implantation failure and recurrent abortion are common indications for endometrial evaluation to determine the implantation window and diagnose endometrial anomalies. There are few research studies comparing the efficacy of different techniques used for endometrial sampling in infertile females during the luteal phase. Likewise, morphometric studies of the endometrium through aspiration biopsy are scant. A cross-sectional study of 30 infertile and 10 fertile females was carried out. The study participants underwent hysteroscopic and aspiration biopsies (pipelle) at the midluteal phase. Computer-assisted morphometric and pathological anatomy analyses were conducted independently by two pathologists blinded to the study. The two endometrial sampling biopsy techniques were compared through morphometric and pathological anatomy analyses using three parameters: a) the amount of material collected for the endometrial studies; b) the scope and origin of sampled materials; and c) the quality of the sample. Both biopsy techniques produced sufficient material for analysis. The directed biopsies yielded higher quality samples from targeted segments of the uterine cavity because samples were homogeneous and had no architectural distortion (p<0.05). Blood was present only in the samples obtained through a Pipelle. Endometritis was detected in 10% of the infertile women. Our findings suggest that hysteroscopic biopsies are superior to blinded aspiration biopsies
- PublicationOpen AccessThe regulation mechanism of apoptosis by visfatin in the mesenteric lymph nodes of LPS-treated rats(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Xiao, Ke; Zhou, Ying; Yuan, Huai Rui; Cui, Lu; Rehman, Zia ur; Ansari, Abdur Rahman; Yang, Zhi; Peng, Ke Mei; Song, HuiVisfatin is an adipocytokine displaying multiple functional properties, which plays a role in the regulation of cell apoptosis and inflammation by an as yet unidentified mechanism. The aim of the present study was to determine if visfatin is involved in apoptosis pathway induced by LPS in rat Mesenteric lymph nodes (MLNs). Experimental rats were divided into four groups and MLNs samples were collected from each group. The morphological changes of the MLNs were examined by histological imaging. CD68 and ENPP1 were detected with immunohistochemistry and Western Blot. Apoptosis was evaluated with TUNEL and Flow Cytometry, the mRNA levels of the apoptosisrelated genes were detected by qRT-PCR, and the protein levels of the apoptotic-related factors were detected by western blot. The main results showed that visfatin could significantly increase the macrophages in MLNs and prevent cell apoptosis from LPS-induced mesenteric lymph nodes, activate apoptotic signaling pathways and regulate the mRNA levels of the apoptosis-related genes. Visfatin had a pro-apoptotic effect on normal MLNs, whereas it exerted an anti-apoptotic effect during LPSinduced cell apoptosis in rat MLNs. In short, visfatin plays a dual role in the apoptosis in rat MLNs, which is mediated by both the mitochondrial apoptotic pathway and the death-receptor apoptotic pathway.
- PublicationOpen AccessAltered expression of human endogenous retroviruses syncytin-1, syncytin-2 and their receptors in human normal and gestational diabetic placenta(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Soygur, Bikem; Sati, Leyla; Demir, RamazanIntroduction: Syncytins belong to the Human Endogenous Retrovirus family. The syncytin-1 receptor, SLC1A5, and syncytin-2 receptor, MFSD2, interact with their respective syncytin proteins to induce syncytiotrophoblast formation. However, there is no information about syncytins in gestational diabetic placenta. Therefore, we studied the expression and localization of syncytins and their receptors during normal placental development and in gestational diabetic placenta. Methods: Immunohistochemistry and Western-blot methods were performed with antibodies against syncytin-1, syncytin-2, SLC1A5 and MFSD2 in human first trimester placental tissues, normal term and gestational diabetic placentas. Syncytin-1, syncytin-2 and MFSD2 mRNA transcripts were determined by qRTPCR in normal and diabetic term placentas. Results: Cytoplasmic syncytin-1, syncytin-2, SLC1A5 and MFSD2 immunoreactions were observed in the trophoblastic layers in all placental samples. Some of the stromal cells showed strong cytoplasmic punctate staining. There were significantly weak syncytin-2 and MFSD2 immunoreaction intensities in diabetic placentas by ImageJ analysis, in parallel with decreased syncytin-2 and MFSD2 proteins in diabetic placentas by Westernblot. Protein expression of SLC1A5 increased dramatically in early pregnancy compared to term placenta. Syncytin-1, syncytin-2 and MFSD2 mRNA transcripts showed similar relative expression pattern by qRT-PCR. Discussion: Syncytins were localized not only in cytotrophoblast cells and the basement membrane of the syncytiotrophoblast but also in the apical microvillous membrane, cytoplasm of syncytiotrophoblast, some of the stromal cells and endothelium. Decreased syncytin-2 and MFSD2 proteins in gestational diabetic placentas might cause abnormal syncytiotrophoblast formation and possibly be involved in the pathology. Therefore, our study highlights an important potential relationship between syncytins and gestational diabetic placenta.
- PublicationOpen AccessTolerogenic responses of CD206+, CD83+, FOXP3+, and CTLA-4 to sericin/polyvinyl alcohol/glycerin scaffolds relevant to IL-33 and HSP60 activity(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Ampawong, Sumate; Aramwit, PornanongSilk sericin-releasing (sericin/polyvinyl alcohol (PVA)/glycerin) scaffolds have been designed for wound dressing applications using different fabrication techniques that influence scaffold antigenicity. The immunological tolerance of scaffolds depends on the balance of immunogenic and tolerogenic responses modulated by dendritic cells (DCs). An in vivo skin implantation model was used to compare the tolerogenic effect of sericin/PVA/glycerin scaffolds prepared by freeze-drying versus salt-leaching techniques, using an Allevyn® scaffold as a control. Immunohistochemical and histopathological studies were performed to evaluate tolerogenic DCs (CD206+), immunogenic DCs (CD83+), regulatory T-cells (FOXP3+ and CTLA-4), a proinflammatory cytokine (interleukin 33: IL-33), a stress marker (heat shock protein 60; HSP60), histopathological changes and related inflammatory cells. It was found that both sericin/PVA/ glycerin scaffolds were tolerogenic and induced early activated Treg functions, while the Allevyn® scaffold was immunogenic. However, the tolerance of the freezedried sericin/PVA/glycerin scaffolds was not as consistent as the salt-leached sericin/PVA/glycerin scaffolds, indicated by the low level of CTLA-4 expression. This was probably due to molecular crosslinking and the morphological and mechanical properties of the freeze-drying technique, which would enhance the immune response. Severe inflammatory responses (including mast cell degranulation and foreign body giant cell accumulation) and histopathological changes (including fat infiltration and fibrosis formation) were mainly found with the Allevyn® scaffold, presumably from its architecture and chemical composition, especially polyurethane. The up-regulation of IL-33 and HSP60 with the Allevyn® scaffold was correlated with the inflammatory and pathological levels. Our findings suggested that salt-leached sericin/PVA/glycerin scaffolds were tolerogenic, induced a low inflammatory response and were appropriate for wound dressing applications.
- PublicationOpen AccessTWEAK/Fn14 signaling: a promising target in intervertebral disc degeneration(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Liu, Yu-Ping; Yuan, Chong-Ming; Zhang, Shuai-Gong; Hao, Qing-Hai; Wang, Ming-Ming; Zhang, Zhong; Meng, Qian; Li, Ming; Hao, Yue-Dong
- PublicationOpen AccessMammalian target of rapamycin complex (mTOR) pathway modulates blood-testis barrier (BTB) function through F-actin organization and gap junction(2016) Li, Nan; Yan Cheng, C.mTOR (mammalian target of rapamycin) is one of the most important signaling molecules in mammalian cells which regulates an array of cellular events, ranging from cell metabolism to cell proliferation. Based on the association of mTOR with the core component proteins, such as Raptor (regulatoryassociated protein of mTOR) or Rictor (rapamycinintensive companion of mTOR), mTOR can become the mTORC1 (mammalian target of rapamycin complex 1) or mTORC2, respectively. Studies have shown that during the epithelial cycle of spermatogenesis, mTORC1 promotes remodeling and restructuring of the bloodtestis barrier (BTB) in vitro and in vivo, making the Sertoli cell tight junction (TJ)-permeability barrier “leaky”; whereas mTORC2 promotes BTB integrity, making the Sertoli cell TJ-barrier “tighter”. These contrasting effects, coupled with the spatiotemporal expression of the core signaling proteins at the BTB that confer the respective functions of mTORC1 vs. mTORC2 thus provide a unique mechanism to modulate BTB dynamics, allowing or disallowing the transport of biomolecules and also preleptotene spermatocytes across the immunological barrier. More importantly, studies have shown that these changes to BTB dynamics conferred by mTORC1 and mTORC2 are mediated by changes in the organization of the actin microfilament networks at the BTB, and involve gap junction (GJ) intercellular communication. Since GJ has recently been shown to be crucial to reboot spermatogenesis and meiosis following toxicant-induced aspermatogenesis, these findings thus provide new insightful information regarding the integration of mTOR and GJ to regulate spermatogenesis.
- PublicationOpen AccessExpression of peroxisome proliferator activation receptors (PPARs) and TNFα in placenta tissues in unexplained recurrent pregnancy loss: An immunohistochemical study(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Papamitsou, Theodora; Toskas, Alexandros; Papadopoulou, Kyriaki; Economou, Zinon; Sioga, AntoniaIntroduction. PPAR expression in placenta tissues regulates proinflammatory cytokine production and preserves the quiescence of the uterus during pregnancy. PPAR-γ regulates inflammatory response during gestation while PPAR-δ and TNFα play a central role at implantation, decidualization and placentation. However, their expression levels affect normal pregnancy and may cause gestational complications and miscarriage. The aim of this report is to investigate the relationship of these molecules with unexplained recurrent miscarriage. Materials-methods. The miscarriage group was obtained from 12 women, between the ages of 35 to 42 years, who miscarried during the 1st trimester of gestation and controls consisted of 12 healthy women, between the ages of 27 to 39 years, who had electively terminated their pregnancies, during the 1st trimester of gestation. The abortion material was processed and specimens taken were studied using immunohistochemical methods. Specimens were taken from decidua basalis and decidua parietalis. Monoclonal antibodies were used against PPAR-γ (Peroxisome Proliferator Activation Receptor γ), PPAR-δ and TNFα (Tumor Necrosis Factor alpha). The results were statistically analyzed with Mann-Whitney test. Results. Our research identified PPAR-γ expression in decidua basalis and decidua parietalis from control group and decidua basalis from miscarriage group. PPAR-δ expression was also identified in both deciduas from both groups. Statistically, no significant change in PPAR-γ and PPAR-δ expression was observed between recurrent miscarriage group and controls. On the contrary, a statistically significant upregulation of TNFα was identified in both deciduas between miscarriage group and controls (p<0.05). Conclusions. Our evidence did not support a possible role of PPARs expression in recurrent pregnancy loss. However, a potential involvement of TNFα in the syndrome was reported. Further research should be performed due to insufficient bibliographic data.
- PublicationOpen AccessImmunomodulatory functions of mesenchymal stem cells and possible mechanisms(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Wang, Qing; Ding, Gang; Xu, XinIn addition to their well-studied self-renewal capabilities and multipotent differentiation properties, mesenchymal stem cells (MSCs) have been reported to possess profound immunomodulatory functions both in vitro and in vivo. More and more studies have shown that MSCs are capable of interacting closely with almost all subsets of immune cells, such as T cells, B cells, dendritic cells, natural killer cells, macrophages, and neutrophils etc. The immunomodulatory property of MSCs may shed light on the treatment of a variety of autoimmune and inflammation-related diseases. In this article, we will review the studies on the immunomodulatory and anti-inflammatory functions of MSCs and the mechanisms responsible for the interaction between immune cells and MSCs, which could improve the development of promising approaches for cellmediated immune therapies.