Histology and histopathology Vol.32, nº8 (2017)
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- PublicationOpen AccessIntussusceptive angiogenesis and expression of Tie receptors during porcine metanephric kidney development(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Logothetidou, Anastasia; Vandecasteele, Tim; Van Mulken, Els; Vandevelde, Kimberley; Cornillie, PieterIntussusceptive angiogenesis (IA) is required for normal embryonic vascular development. The Tie family of receptors and their ligands, the angiopoietins, play an important role in the growth or regression of blood vessels which are important not only during development but also throughout an organism’s life. The presence of IA was investigated in glomerular capillaries of the fetal porcine metanephros using Mercox II resin casts. The first signs of IA were observed in stage III glomeruli. Stage IV and V glomeruli showed numerous signs of aligned pillar formation and their successive merging to delineate the vascular entities. Furthermore, immunohistochemistry was used to determine the exact locations of the Tie receptors in the developing porcine metanephric kidneys. Tie1 and Tie2 were found in endothelial cells of all glomeruli. Strong expression of the receptors was found in podocytes of stage V glomeruli whereas a weaker expression was observed in the cuboidal epithelial cells of stage III and IV glomeruli. Remarkably, the receptors were also found in the parietal epithelium of Bowman’s capsule. These findings indicate that there might be an association between Tie receptors and IA during porcine metanephric development and during glomerulogenesis in particular.
- PublicationOpen AccessA modified chemical protocol of decellularization of rat sciatic nerve and its recellularization with mesenchymal differentiated Schwann-Like cells: Morphological and functional assessments(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) García Pérez, M.M.; Martínez Rodríguez, H.G.; López Guerra, G.G.; Soto Domínguez, A.; Said Fernández, S.L.; Morales Ávalos, R.; Elizondo Omaña, R.E.; Montes de Oca Luna, R.; Guzmán López, S.; Castillo Galván, M.L.; Mendoza Lemus, O.F.; Vílchez Cavazos, F.The functional reconstruction of large neural defects usually requires the use of peripheral nerve autografts, though these have certain limitations. As a result, interest in new alternatives for autograft development has risen. The acellular peripheral nerve graft is an alternative for peripheral nerve injury repair, but to date there is not a standardized chemical decellularization method widely accepted. The objective of this study was to propose a modified chemical protocol of decellularization of rat sciatic nerve and its recellularization in vitro with mesenchymal differentiated Schwann-like cells. After the transplantation, an evaluation of its regeneration was performed using morphological and functional tests. The study consisted of two phases; in phase 1, different concentrations and times of exposure of rat sciatic nerves to detergents were tested, to establish a modified chemical protocol for nerve decellularization. The chemical treatment with 3% triton X-100 and 4% sodium deoxycholate for 15 days allowed a complete decellularization whilst conserving the extracellular matrix of the harvested nerve. In phase 2, the decellularized and recellularized alografts were compared against autografts. The morphological analysis showed a higher positivity to specific myelin antibodies in the recellularized group compared to the autograft. There were no differences in this parameter between the control limb and the experimental limb (recellularized group). The functional analysis showed no statistical differences at week 15 in the Sciatic Function Index in the autograft group vs the other groups. This study sets the morphological and functional bases for posterior studies about nerve defects regeneration in humans.
- PublicationOpen AccessDistinct presence of the tight junction protein claudin-3 in olfactory bulb and fila olfactoria of the mouse(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Eppler, Elisabeth; Müller Gerbl, Magdalena; Maly, I. PiotrThe tight junction protein claudin-3 is overexpressed in diverse epithelial tumours and is associated with increased survival, progression and motility of tumour cells. Claudin-3 expression profiles are being increasingly used for diagnostic and prognostic tumour classification. Claudin-3 has been identified as a receptor for Clostridium perfringens enterotoxin, which is under consideration for selective lysis of claudin-3- expressing tumours, particularly brain metastases, and other translational medicine uses. However, the localization of claudin-3 in the brain has not been completely elucidated. While claudin-3 in brain tissue adjacent to claudin-3-expressing metastases had been excluded and low or undetectable levels proposed in the CNS, under physiological conditions, in adult human, rat and mouse brains, claudin-3 was exclusively found in choroid plexus epithelium where it is considered an integral component of the blood-cerebrospinal-fluid barrier. We report here the pronounced presence of claudin-3 not only in the nasal region (as described for rat), but also in the mouse olfactory bulb and nerve using immunohistochemistry and Western blot. Claudin-3 was present in the fila olfactoria from the epithelium to the olfactory nerve and in the main and accessory olfactory bulb. We propose that the abundant presence of claudin3 in the olfactory system, particularly in nerve fibres and the olfactory bulb cone, which we present here, may play a role at the interface of the central and peripheral nervous system, both as barrier and for axonal growth and communication. Thus, claudin-3 should be considered and further explored with regards to treatment approaches addressing the olfactory bulb and nasal region.
- PublicationOpen AccessCorrelation between the high expression levels of cancer-germline genes with clinical characteristics in esophageal squamous cell carcinoma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Chen, Xinfeng; Wang, Liping; Yue, Dongli; Liu, Jinyan; Huang, Lan; Yang, Li; Cao, Ling; Qin, Guohui; Li, Anqi; Wang, Dan; Wang, Meng; Qi, Yu; Zhang, Bin; van der Bruggen, Pierre; Zhang, YiAntigens encoded by cancer-germline genes are attractive targets for cancer immunotherapy. In this study, we aimed to evaluate the mRNA expression of cancer-germline genes, expression of the encoded proteins in patients with esophageal squamous cell carcinoma (ESCC) and their correlations with clinical characteristics. In addition, the effects of downregulation cancer-germline genes on ESCC cells were assessed in vitro. Our results showed that cancer-germline genes were frequently expressed in ESCC samples. The positive rates of in ESCC samples were: 87% of MAGEA3, 60% of MAGE-A4, 65% of MAGE-C2, and 20% of NY-ESO-1 at mRNA level. MAGE-A3 expression was associated with age, lymph node metastasis and tumor stage (all P<0.05), while MAGE-C2 expression was only associated with tumor stage (P<0.05). Furthermore, the MAGE-A3 expressing patients had a poorer overall survival (P<0.05). Multivariate analysis identified MAGE-A3 as an independent poor prognostic marker in ESCC. In vitro assay, ESCC cell lines treated with specific siRNAs to down-regulate MAGE-A3 and MAGE-C2 resulted in decreased colony-formation and migration ability (P<0.05). Epithelial marker E-cadherin was up-regulated in siRNA-MAGE-A3/C2 cells compared to controls, whereas mesenchymal markers Vimentin, N-cadherin and Slug were downregulated (all P<0.05), suggesting a role for MAGE-A3/C2 in ESCC metastasis through inducing epithelial-mesenchymal transition. The present study revealed that cancergermline genes and their encoded proteins were frequently expressed in ESCC tumor samples and were related to poor prognosis. Thus, cancer-germline genes may serve as useful biomarkers and potential targets for ESCC patients
- PublicationOpen AccessPuerarin and zinc additively prevent mandibular bone loss through inhibiting osteoclastogenesis in ovariectomized rats(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Liu, Hao; Li, Wei; Jia, Shengnan; Li, BinbinPuerarin and zinc play a key role in preventing osteoporotic-related bone loss. Previous research on puerarin or zinc mainly focused on the antiosteoporotic effects of long bone. However, it is obscure for puerarin or zinc to prevent mandibular osteoporosis. Here, we explore the effects on additive coadministration of puerarin and zinc on preventing mandibular bone loss in ovariectomized rats, and evaluate the underlying mechanisms ex vivo. Rats were ovariectomized and administrated puerarin, zinc or both. After 12 weeks, bone mineral density (BMD) and histomorphometry of mandibles were measured by micro-CT. The mechanical properties were determined using a threepoint bending test. Then, osteogenic differentiation of primary bone marrow stromal cells (BMSCs) and osteoclastogenesis of bone marrow mononuclear were performed ex vivo. The culture supernatant and serum level of bone biochemical markers including osteoprotegerin (OPG), osteopontin (OPN), receptor activator of nuclear factor (NF)-κB ligand (RANKL), and tartrate-resistant acid phosphatase (TRAP) were detected by ELISA. Culture supernatant and serum levels of calcium were measured using a Plasma Emission Spectrometer. One-way ANOVA was used for statistical analyses. The results showed that administration of puerarin plus zinc prevented the decrease in mandibular BMD and bone morphometrical parameters more effectively than single use of puerarin or zinc (p<0.05), which was similar to the biomechanical tests (p<0.05). Furthermore, puerarin and zinc additively up-regulated OPG, OPN protein levels, Ca ion level and down-regulated RANKL, TRAP protein levels. In conclusion, puerarin and zinc additively prevent mandibular bone loss through inhibiting osteoclastogenesis in ovariectomized rats, which will shed more light on the potential use of puerarin and zinc in the prevention/treatment of oral bone loss clinically.