scholarly journals Ex Vivo and in Vivo Study of Sucrosomial® Iron Intestinal Absorption and Bioavailability

2018 ◽  
Vol 19 (9) ◽  
pp. 2722 ◽  
Author(s):  
Angela Fabiano ◽  
Elisa Brilli ◽  
Letizia Mattii ◽  
Lara Testai ◽  
Stefania Moscato ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Laser Scanning ◽  
Time Course ◽  
Ex Vivo ◽  
Iron Bioavailability ◽  
Laser Scanning Microscopy ◽  
Raw Material ◽  
Confocal Laser ◽  
Ussing Chambers

The present study aimed to demonstrate that Sideral® RM (SRM, Sucrosomial® Raw Material Iron) is transported across the excised intestine via a biological mechanism, and to investigate the effect that this transport route may produce on oral iron absorption, which is expected to reduce the gastrointestinal (GI) side effects caused by the bioavailability of non-absorbed iron. Excised rat intestine was exposed to fluorescein isothiocyanate (FITC)-labeled SRM in Ussing chambers followed by confocal laser scanning microscopy to look for the presence of fluorescein-tagged vesicles of the FITC-labeled SRM. To identify FITC-labeled SRM internalizing cells, an immunofluorescence analysis for macrophages and M cells was performed using specific antibodies. Microscopy analysis revealed the presence of fluorescein positive particulate structures in tissues treated with FITC-labeled SRM. These structures do not disintegrate during transit, and concentrate in macrophage cells. Iron bioavailability was assessed by determining the time-course of Fe3+ plasma levels. As references, iron contents in liver, spleen, and bone marrow were determined in healthy rats treated by gavage with SRM or ferric pyrophosphate salt (FP). SRM significantly increased both area under the curve (AUC) and clearance maxima (Cmax) compared to FP, thus increasing iron bioavailability (AUCrel = 1.8). This led to increased iron availability in the bone marrow at 5 h after single dose gavage.

The fate of medial edge epithelial cells during palatal fusion in vitro: an analysis by DiI labelling and confocal microscopy

Development ◽  
1992 ◽  
Vol 114 (2) ◽  
pp. 379-388 ◽  
Author(s):  
M.J. Carette ◽  
M.W. Ferguson
Keyword(s):  
Laser Scanning ◽  
Time Course ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Palatal Fusion ◽  
Morphological Criteria ◽  
Medial Edge ◽  
Mesenchymal Transformation

Fusion of bilateral shelves, to form the definitive mammalian secondary palate, is critically dependent on removal of the medial edge cells that constitute the midline epithelial seam. Conflicting views suggest that programmed apoptotic death or epithelial-mesenchymal transformation of these cells is predominantly involved. Due in part to the potentially ambiguous interpretation of static images and the notable absence of fate mapping studies, the process by which this is achieved has, however, remained mechanistically equivocal. Using an in vitro mouse model, we have selectively labelled palatal epithelia with DiI and examined the fate of medial edge epithelial (MEE) cells during palatal fusion by localisation using a combination of conventional histology and confocal laser scanning microscopy (CLSM). In dynamic studies using CLSM, we have made repetitive observations of the same palatal cultures in time-course investigations. Our results concurred with the established morphological criteria of seam degeneration; however, they provided no evidence of MEE cell death or transformation. Instead we report that MEE cells migrate nasally and orally out of the seam and are recruited into, and constitute, epithelial triangles on both the oral and nasal aspects of the palate. Subsequently these cells become incorporated into the oral and nasal epithelia on the surface of the palate. We hypothesize an alternative method of seam degeneration in vivo which largely conserves the MEE population by recruiting it into the nasal and oral epithelia.

scholarly journals Clinical Applications of In Vivo and Ex Vivo Confocal Microscopy

2021 ◽  
Vol 11 (5) ◽  
pp. 1979
Author(s):  
Stefania Guida ◽  
Federica Arginelli ◽  
Francesca Farnetani ◽  
Silvana Ciardo ◽  
Laura Bertoni ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Laser Scanning ◽  
Ex Vivo ◽  
Real Life ◽  
Laser Scanning Microscopy ◽  
Pathological Examination ◽  
Confocal Laser ◽  
Fluorescence Confocal Microscopy ◽  
Skin Cancers

Confocal laser scanning microscopy (CLSM) has been introduced in clinical settings as a tool enabling a quasi-histologic view of a given tissue, without performing a biopsy. It has been applied to many fields of medicine mainly to the skin and to the analysis of skin cancers for both in vivo and ex vivo CLSM. In vivo CLSM involves reflectance mode, which is based on refractive index of cell structures serving as endogenous chromophores, reaching a depth of exploration of 200 μm. It has been proven to increase the diagnostic accuracy of skin cancers, both melanoma and non-melanoma. While histopathologic examination is the gold standard for diagnosis, in vivo CLSM alone and in addition to dermoscopy, contributes to the reduction of the number of excised lesions to exclude a melanoma, and to improve margin recognition in lentigo maligna, enabling tissue sparing for excisions. Ex vivo CLSM can be performed in reflectance and fluorescent mode. Fluorescence confocal microscopy is applied for “real-time” pathological examination of freshly excised specimens for diagnostic purposes and for the evaluation of margin clearance after excision in Mohs surgery. Further prospective interventional studies using CLSM might contribute to increase the knowledge about its application, reproducing real-life settings.

scholarly journals Design and Engineering of “Green” Nanoemulsions for Enhanced Topical Delivery of Bakuchiol Achieved in a Sustainable Manner: A Novel Eco-Friendly Approach to Bioretinol

2021 ◽  
Vol 22 (18) ◽  
pp. 10091
Author(s):  
Agnieszka Lewińska ◽  
Marta Domżał-Kędzia ◽  
Ewa Maciejczyk ◽  
Marcin Łukaszewicz ◽  
Urszula Bazylińska
Keyword(s):  
Human Skin ◽  
Laser Scanning ◽  
Ex Vivo ◽  
Rapeseed Meal ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Biological Investigation ◽  
Vis Spectroscopy

In the present work, we establish novel “environmentally-friendly” oil-in-water nanoemulsions to enhance the transdermal delivery of bakuchiol, the so-called “bioretinol” obtained from powdered Psoralea corylifolia seeds via a sustainable process, i.e., using a supercritical fluid extraction approach with pure carbon dioxide (SC-CO2). According to Green Chemistry principles, five novel formulations were stabilized by “green” hybrid ionic surfactants such as coco-betaine—surfactin molecules obtained from coconut and fermented rapeseed meal. Preliminary optimization studies involving three dispersion stability tests, i.e., centrifugation, heating, and cooling cycles, indicated the most promising candidates for further physicochemical analysis. Finally, nanoemulsion colloidal characterization provided by scattering (dynamic and electrophoretic light scattering as well as backscattering), microscopic (transmission electron and confocal laser scanning microscopy), and spectroscopic (UV–Vis spectroscopy) methods revealed the most stable nanocarrier for transdermal biological investigation. In vitro, topical experiments provided on human skin cell line HaCaT keratinocytes and normal dermal NHDF fibroblasts indicated high cell viability upon treatment of the tested formulation with a final 0.02–0.2 mg/mL bakuchiol concentration. This excellent biocompatibility was confirmed by ex vivo and in vivo tests on animal and human skin tissue. The improved permeability and antiaging potential of the bakuchiol-encapsulated rich extract were observed, indicating that the obtained ecological nanoemulsions are competitive with commercial retinol formulations.

scholarly journals Meibomian Glands or Not? Identification of In Vivo and Ex Vivo Confocal Microscopy Features and Histological Correlates in the Eyelid Margin

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Yu-jing Wang ◽  
Min Ke
Keyword(s):  
Confocal Microscopy ◽  
Laser Scanning ◽  
Ex Vivo ◽  
Meibomian Gland ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Eyelid Margin ◽  
Healthy Humans ◽  
Skin Appendages

Purpose. In vivo confocal laser scanning microscopy (CLSM) is an emerging diagnostic tool allowing fast and easy microscopic tissue examination. For the diagnostics of pathological eyelid margin lesions, the knowledge of the normal eyelid margin is essential. Methods. We examined 18 eyelid margins of healthy humans using the in vivo CLSM device and 10 samples of healthy eyelid margins from donor sites with ex vivo CLSM and compared the findings to the corresponding histological sections of donor sites. Cross-section images of different depths and depths of different skin appendages were measured. Results. The depth observed by in vivo CLSM is less than 150 μm into the eyelid. Images of the epidermis and superficial dermis skin, appendages including hair follicle, and sebaceous catheters can be captured associated with histopathology and ex vivo confocal microscopy. In correlation with histopathology, we identified different layers of the eyelid margin, different layers of the epidermis, and skin appendages by ex vivo confocal microscopy. Conclusions. The study offers an overview of the in vivo confocal microscopy human eyelid margin characteristics in comparison to the standard histological examination and confirms that in vivo CLSM could not observe the meibomian gland acini structure.

Vacuoles Induced in Mammalian Cells by Helicobacter Cytotoxin are Acidic Organelles

1990 ◽  
Vol 48 (3) ◽  
pp. 168-169
Author(s):  
M. H. Chestnut ◽  
C. E. Catrenich
Keyword(s):  
Acridine Orange ◽  
Mammalian Cells ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ulcer Disease ◽  
Gastroduodenal Disease ◽  
H Pylori

Helicobacter pylori is a non-invasive, Gram-negative spiral bacterium first identified in 1983, and subsequently implicated in the pathogenesis of gastroduodenal disease including gastritis and peptic ulcer disease. Cytotoxic activity, manifested by intracytoplasmic vacuolation of mammalian cells in vitro, was identified in 55% of H. pylori strains examined. The vacuoles increase in number and size during extended incubation, resulting in vacuolar and cellular degeneration after 24 h to 48 h. Vacuolation of gastric epithelial cells is also observed in vivo during infection by H. pylori. A high molecular weight, heat labile protein is believed to be responsible for vacuolation and to significantly contribute to the development of gastroduodenal disease in humans. The mechanism by which the cytotoxin exerts its effect is unknown, as is the intracellular origin of the vacuolar membrane and contents. Acridine orange is a membrane-permeant weak base that initially accumulates in low-pH compartments. We have used acridine orange accumulation in conjunction with confocal laser scanning microscopy of toxin-treated cells to begin probing the nature and origin of these vacuoles.

scholarly journals Alien Wood Species as a Resource for Wood-Plastic Composites

2020 ◽  
Vol 11 (1) ◽  
pp. 44
Author(s):  
Sergej Medved ◽  
Daša Krapež Tomec ◽  
Angela Balzano ◽  
Maks Merela
Keyword(s):  
Wood Species ◽  
Dimensional Stability ◽  
Laser Scanning ◽  
Large Scale ◽  
Laser Scanning Microscopy ◽  
Raw Material ◽  
Confocal Laser ◽  
Alien Plant ◽  
Wood Plastic Composites ◽  
Plastic Composites

Since invasive alien species are one of the main causes of biodiversity loss in the region and thus of changes in ecosystem services, it is important to find the best possible solution for their removal from nature and the best practice for their usability. The aim of the study was to investigate their properties as components of wood-plastic composites and to investigate the properties of the wood-plastic composites produced. The overall objective was to test the potential of available alien plant species as raw material for the manufacture of products. This would contribute to sustainability and give them a better chance of ending their life cycle. One of the possible solutions on a large scale is to use alien wood species for the production of wood plastic composites (WPC). Five invasive alien hardwood species have been used in combination with polyethylene powder (PE) and maleic anhydride grafted polyethylene (MAPE) to produce various flat pressed WPC boards. Microstructural analyses (confocal laser scanning microscopy and scanning electron microscopy) and mechanical tests (flexural strength, tensile strength) were performed. Furthermore, measurements of density, thickness swelling, water absorption and dimensional stability during heating and cooling were carried out. Comparisons were made between the properties of six WPC boards (five alien wood species and mixed boards). The results showed that the differences between different invasive alien wood species were less obvious in mechanical properties, while the differences in sorption properties and dimensional stability were more significant. The analyses of the WPC structure showed a good penetration of the polymer into the lumens of the wood cells and a fine internal structure without voids. These are crucial conditions to obtain a good, mechanically strong and water-resistant material.

scholarly journals Targeted Liposomal Chemotherapies to Treat Triple-Negative Breast Cancer

Cancers ◽  
2021 ◽  
Vol 13 (15) ◽  
pp. 3749
Author(s):  
Yingnan Si ◽  
Ya Zhang ◽  
Hanh Giai Ngo ◽  
Jia-Shiung Guan ◽  
Kai Chen ◽  
...  
Keyword(s):  
Targeted Delivery ◽  
Laser Scanning ◽  
Triple Negative ◽  
Imaging System ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Surface Binding ◽  
Synthesis Inhibitor ◽  
Tnbc Cell

Triple-negative breast cancers (TNBCs) are highly aggressive and recurrent. Standard cytotoxic chemotherapies are currently the main treatment options, but their clinical efficacies are limited and patients usually suffer from severe side effects. The goal of this study was to develop and evaluate targeted liposomes-delivered combined chemotherapies to treat TNBCs. Specifically, the IC50 values of the microtubule polymerization inhibitor mertansine (DM1), mitotic spindle assembly defecting taxane (paclitaxel, PTX), DNA synthesis inhibitor gemcitabine (GC), and DNA damage inducer doxorubicin (AC) were tested in both TNBC MDA-MB-231 and MDA-MB-468 cells. Then we constructed the anti-epidermal growth factor receptor (EGFR) monoclonal antibody (mAb) tagged liposomes and confirmed its TNBC cell surface binding using flow cytometry, internalization with confocal laser scanning microscopy, and TNBC xenograft targeting in NSG female mice using In Vivo Imaging System. The safe dosage of anti-EGFR liposomal chemotherapies, i.e., <20% body weight change, was identified. Finally, the in vivo anti-tumor efficacy studies in TNBC cell line-derived xenograft and patient-derived xenograft models revealed that the targeted delivery of chemotherapies (mertansine and gemcitabine) can effectively inhibit tumor growth. This study demonstrated that the targeted liposomes enable the new formulations of combined therapies that improve anti-TNBC efficacy.

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