scholarly journals Haemophilin-Producing Strains of Haemophilus haemolyticus Protect Respiratory Epithelia from NTHi Colonisation and Internalisation

Pathogens ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 29
Author(s):  
Brianna Atto ◽  
Dale Kunde ◽  
David A Gell ◽  
Stephen Tristram
Keyword(s):  
Inhibitory Activity ◽  
Therapeutic Potential ◽  
Epithelial Cell Line ◽  
The Novel ◽  
Nontypeable Haemophilus Influenzae ◽  
Respiratory Epithelia ◽  
Culture Models ◽  
Host Cell Interactions ◽  
Cell Culture Models

Nontypeable Haemophilus influenzae (NTHi) is a significant respiratory tract pathogen responsible for infections that collectively pose a substantial health and socioeconomic burden. The clinical course of these infections is largely dictated by NTHi interactions with host respiratory epithelia, and thus, approaches that disrupt colonisation and invasion may have significant therapeutic potential. Survival, successful host–cell interactions, and pathogenesis are reliant on NTHi’s ability to sequester host-derived haem. Previously, we demonstrated the therapeutic potential of exploiting this haem-dependence using a closely related competitor bacterium, Haemophilus haemolyticus (Hh). Hh strains capable of producing the novel haem-binding protein haemophilin (Hpl) possessed potent inhibitory activity by restricting NTHi access to haem in a broth co-culture environment. Here, we extend this work to cell culture models that more closely represent the human respiratory epithelium and show that Hh strains with high levels of hpl expression protect epithelial cell line monolayers against adhesion and invasion by NTHi. Inhibitory activity was dependent on the level of Hpl production, which was stimulated by NTHi challenge and nasopharyngeal cell exposure. Provided these protective benefits translate to in vivo applications, Hpl-producing Hh may have probiotic utility against NTHi infections by inhibiting requisite nasopharyngeal colonisation.

scholarly journals Dynamics of Internalization and Intracellular Interaction of Tau Antibodies and Human Pathological Tau Protein in a Human Neuron-Like Model

2020 ◽  
Vol 11 ◽  
Author(s):  
Dov B. Shamir ◽  
Yan Deng ◽  
Qian Wu ◽  
Swananda Modak ◽  
Erin E. Congdon ◽  
...  
Keyword(s):  
Tau Protein ◽  
Therapeutic Potential ◽  
Ex Vivo ◽  
Human Model ◽  
Paired Helical Filament ◽  
Isotype Control ◽  
Culture Models ◽  
Cellular Compartments ◽  
Cell Culture Models

We and others have shown in various in vivo, ex vivo and cell culture models that several tau antibodies interact with pathological tau within neurons. To further clarify this interaction in a dynamic human model, we differentiated SH-SY5Y cells with retinoic acid and BDNF to create a neuron-like model. Therein, tau antibodies were primarily taken up by receptor-mediated endocytosis, and prevented toxicity of human brain-derived paired helical filament-enriched tau (PHF). Subsequently, we monitored in real-time the interaction of antibodies and PHF within endocytic cellular compartments. Cells were pre-treated with fluorescently-tagged PHF and then incubated with tau antibodies, 4E6, 6B2, or non-specific isotype control IgG1 labeled with a pH sensitive dye. The uptake and binding of the efficacious antibody, 4E6, to PHF occurred mainly within the soma, whereas the ineffective antibody, 6B2, and ineffective control IgG1, were visualized via the processes and showed limited colocalization with PHF within this period. In summary, we have developed a neuron-like model that clarifies the early intracellular dynamics of the interaction of tau antibodies with pathological tau, and identifies features associated with efficacy. Since the model is entirely human, it is suitable to verify the therapeutic potential of humanized antibodies prior to extensive clinical trials.

scholarly journals The Communication between Ocular Surface and Nasal Epithelia in 3D Cell Culture Technology for Translational Research: A Narrative Review

2021 ◽  
Vol 22 (23) ◽  
pp. 12994
Author(s):  
Malik Aydin ◽  
Jana Dietrich ◽  
Joana Witt ◽  
Maximiliane S. C. Finkbeiner ◽  
Jonas J.-H. Park ◽  
...  
Keyword(s):  
Cell Culture ◽  
Nasal Cavity ◽  
Ocular Surface ◽  
Measles Virus ◽  
3D Culture ◽  
3D Cell Culture ◽  
Narrative Review ◽  
Culture Models ◽  
Cell Culture Models

There is a lack of knowledge regarding the connection between the ocular and nasal epithelia. This narrative review focuses on conjunctival, corneal, ultrastructural corneal stroma, and nasal epithelia as well as an introduction into their interconnections. We describe in detail the morphology and physiology of the ocular surface, the nasolacrimal ducts, and the nasal cavity. This knowledge provides a basis for functional studies and the development of relevant cell culture models that can be used to investigate the pathogenesis of diseases related to these complex structures. Moreover, we also provide a state-of-the-art overview regarding the development of 3D culture models, which allow for addressing research questions in models resembling the in vivo situation. In particular, we give an overview of the current developments of corneal 3D and organoid models, as well as 3D cell culture models of epithelia with goblet cells (conjunctiva and nasal cavity). The benefits and shortcomings of these cell culture models are discussed. As examples for pathogens related to ocular and nasal epithelia, we discuss infections caused by adenovirus and measles virus. In addition to pathogens, also external triggers such as allergens can cause rhinoconjunctivitis. These diseases exemplify the interconnections between the ocular surface and nasal epithelia in a molecular and clinical context. With a final translational section on optical coherence tomography (OCT), we provide an overview about the applicability of this technique in basic research and clinical ophthalmology. The techniques presented herein will be instrumental in further elucidating the functional interrelations and crosstalk between ocular and nasal epithelia.

Characterizing the Phenotypic Response of Endothelial Cells Cultured on Pro-Angiogenic In Vitro Tumor Mimics

2012 ◽  
Author(s):  
Christopher S. Szot ◽  
Cara F. Buchanan ◽  
Joseph W. Freeman ◽  
Marissa Nichole Rylander
Keyword(s):  
Cancer Research ◽  
Cancer Progression ◽  
Tumor Development ◽  
New Drugs ◽  
Preclinical Models ◽  
Fda Approval ◽  
Culture Models ◽  
Cell Culture Models

Despite the 200 billion dollars invested in cancer therapy research and development since 1971, only 5% of new drugs entering clinical trials successfully obtain FDA approval [1, 2]. There is a growing concern in the cancer research community that this slow movement in progress stems from the need for improved preclinical models for testing new therapeutic agents [1]. A burgeoning interface between cancer research and tissue engineering is transforming how tumor development is being studied in vitro. As a result, complex 3D cancer cell culture models are beginning to be developed with phenotypes representative of in vivo cancer progression [3].

scholarly journals PPAR-gamma Fun(gi) With Prostaglandin

2020 ◽  
Vol 17 ◽  
pp. 155076291989964
Author(s):  
Robert J. Evans ◽  
Simon A. Johnston
Keyword(s):  
Partial Agonist ◽  
Ppar Gamma ◽  
Host Cells ◽  
Culture Models ◽  
Important Regulator ◽  
Inflammatory Phenotypes ◽  
First Time ◽  
Cell Culture Models

In our recent publication, we show for the first time that the fungal pathogen Cryptococcus neoformans is able to manipulate host cells by producing eicosanoids that mimic those found in the host. Using complementary in vivo zebrafish and in vitro macrophage cell culture models of Cryptococcus infection, we found that these eicosanoids manipulate host innate immune cells by activating the host receptor PPAR-gamma which is an important regulator of macrophage inflammatory phenotypes. We initially identified PGE2 as the eicosanoid species responsible for this effect; however, we later found that a derivative of PGE2—15-keto-PGE2—was ultimately responsible and that this eicosanoid acted as a partial agonist to PPAR-gamma. In this commentary, we will discuss some of the concepts and conclusions in our original publication and expand on their implications and future directions.

scholarly journals Optimizations of In Vitro Mucus and Cell Culture Models to Better Predict In Vivo Gene Transfer in Pathological Lung Respiratory Airways: Cystic Fibrosis as an Example

Pharmaceutics ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 47
Author(s):  
Rosy Ghanem ◽  
Véronique Laurent ◽  
Philippe Roquefort ◽  
Tanguy Haute ◽  
Sophie Ramel ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Gene Therapy ◽  
Cell Culture ◽  
Gene Transfer ◽  
Gene Delivery ◽  
Culture Models ◽  
Transfer Strategies ◽  
Cell Culture Models

The respiratory epithelium can be affected by many diseases that could be treated using aerosol gene therapy. Among these, cystic fibrosis (CF) is a lethal inherited disease characterized by airways complications, which determine the life expectancy and the effectiveness of aerosolized treatments. Beside evaluations performed under in vivo settings, cell culture models mimicking in vivo pathophysiological conditions can provide complementary insights into the potential of gene transfer strategies. Such models must consider multiple parameters, following the rationale that proper gene transfer evaluations depend on whether they are performed under experimental conditions close to pathophysiological settings. In addition, the mucus layer, which covers the epithelial cells, constitutes a physical barrier for gene delivery, especially in diseases such as CF. Artificial mucus models featuring physical and biological properties similar to CF mucus allow determining the ability of gene transfer systems to effectively reach the underlying epithelium. In this review, we describe mucus and cellular models relevant for CF aerosol gene therapy, with a particular emphasis on mucus rheology. We strongly believe that combining multiple pathophysiological features in single complex cell culture models could help bridge the gaps between in vitro and in vivo settings, as well as viral and non-viral gene delivery strategies.

scholarly journals Selenomethionine-Dominated Selenium-Enriched Peanut Protein Ameliorates Alcohol-Induced Liver Disease in Mice by Suppressing Oxidative Stress

Foods ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2979
Author(s):  
Lin Gao ◽  
Jiawei Yuan ◽  
Yuhuan Cheng ◽  
Mengling Chen ◽  
Genhua Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Liver Disease ◽  
Therapeutic Agents ◽  
Peanut Protein ◽  
Culture Models ◽  
Therapeutic Properties ◽  
Cell Culture Models ◽  
Dependent Mechanism

Numerous natural compounds are considered as potential therapeutic agents against alcohol-induced liver disease (ALD). Research shows that selenium (Se) has a variety of bioactivities, including liver protecting ability. The present study based on in vitro cell culture models and in vivo mouse models was aimed at examining the contribution of selenomethionine (SeMet)-dominated Se-enriched peanut protein (SePP) to liver protection. SeMet and especially SePP reversed cell viability and cell death, inhibited ethanol induced CYP2E1 activation, decreased reactive oxygen species level, and restored GSH level. Hence, SeMet-dominated SePP alleviates alcohol-induced AML-12 cytotoxicity by suppressing oxidative stress. The p38-dependent mechanism was found to be responsible for SePP-induced Nrf-2 activation. Furthermore, supplementation with SePP and SeMet regulated lipid metabolism and reduced oxidative stress, minimizing liver damage in mice. Selenomethionine-dominated SePP possesses potential therapeutic properties and can be used to treat ALD through the suppression of oxidative stress.

scholarly journals A biomimetic liver model recapitulating bio-physical properties and tumour stroma interactions in hepatocellular carcinoma

2020 ◽  
Author(s):  
C. Calitz ◽  
N. Pavlovic ◽  
J. Rosenquist ◽  
C. Zagami ◽  
A. Samanta ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Culture ◽  
Liver Disease ◽  
Physical Properties ◽  
Chronic Liver Disease ◽  
3D Model ◽  
Stromal Compartment ◽  
Culture Models ◽  
Cell Culture Models

ABSTRACTHepatocellular carcinoma (HCC) is a primary liver tumor developing in the wake of chronic liver disease. Chronic liver disease and inflammation leads to a fibrotic environment actively supporting and driving hepatocarcinogenesis. Insight into hepatocarcinogenesis in terms of the interplay between the tumor stroma micro-environment and tumor cells is thus of considerable importance. Three-dimensional (3D) cell culture models are proposed as the missing link between current in vitro 2D cell culture models and in vivo animal models. Our aim was to design a novel 3D biomimetic HCC model with accompanying fibrotic stromal compartment and vasculature. Physiologically relevant hydrogels such as collagen and fibrinogen were incorporated to mimicking the bio-physical properties of the tumor ECM. In our model LX2 and HepG2 cells embedded in a hydrogel matrix were seeded onto the inverted insert membrane of a Transwell™ system. HUVEC cells were then seeded onto the opposite side of the membrane. Three formulations consisting of ECM-hydrogels embedded with cells were prepared and the bio-physical properties determined by rheology. Cell viability was determined by the AlamarBlue® assay over 21-days. The effect of the chemotherapeutic drug doxorubicin was evaluated in both a 2D co-culture and our 3D model for a period of 72h. We show that this model is viable for 25-days and gives rise to metastatic tumor nodules after 17 days in culture. Rheology results show that bio-physical properties of a fibrotic, cirrhotic and HCC liver can be successfully mimicked. Overall, results indicate that this 3D model is more representative of the in vivo situation compared to traditional 2D cultures. Our 3D tumor model showed a decreased response to chemotherapeutics, mimicking drug resistance typically seen in HCC patients. This model could in future provide a valuable new platform to study multifocal HCC or to identify mechanisms that contribute to early stages of metastasis.SUMMARYA protocol for a novel 3D biomimetic HCC model with accompanying fibrotic stromal compartment and vasculature, to study endocrine and paracrine signaling in liver cancer. The model uses physiological relevant hydrogels in ratios mimicking the bio-physical properties of the stromal extracellular matrix, which is an active mediator of cellular interactions, tumor growth and metastasis.

scholarly journals In Vivo Airway Surface Liquid Cl− Analysis with Solid-State Electrodes

2001 ◽  
Vol 119 (1) ◽  
pp. 3-14 ◽  
Author(s):  
Ray A. Caldwell ◽  
Barbara R. Grubb ◽  
Robert Tarran ◽  
Richard C. Boucher ◽  
Michael R. Knowles ◽  
...  
Keyword(s):  
Solid State ◽  
Airway Surface Liquid ◽  
Culture Models ◽  
Normal Human ◽  
Surface Liquid ◽  
Multiple Species ◽  
Cell Culture Models ◽  
Made In

The pathogenesis of cystic fibrosis (CF) airways disease remains controversial. Hypotheses that link mutations in CFTR and defects in ion transport to CF lung disease predict that alterations in airway surface liquid (ASL) isotonic volume, or ion composition, are critically important. ASL [Cl−] is pivotal in discriminating between these hypotheses, but there is no consensus on this value given the difficulty in measuring [Cl−] in the “thin” ASL (∼30 μm) in vivo. Consequently, a miniaturized solid-state electrode with a shallow depth of immersion was constructed to measure ASL [Cl−] in vivo. In initial experiments, the electrode measured [Cl−] in physiologic salt solutions, small volume (7.6 μl) test solutions, and in in vitro cell culture models, with ≥93% accuracy. Based on discrepancies in reported values and/or absence of data, ASL Cl− measurements were made in the following airway regions and species. First, ASL [Cl−] was measured in normal human nasal cavity and averaged 117.3 ± 11.2 mM (n = 6). Second, ASL [Cl−] measured in large airway (tracheobronchial) regions were as follows: rabbit trachea and bronchus = 114.3 ± 1.8 mM; (n = 6) and 126.9 ± 1.7 mM; (n = 3), respectively; mouse trachea = 112.8 ± 4.2 mM (n = 13); and monkey bronchus = 112.3 ± 10.9 mM (n = 3). Third, Cl− measurements were made in small (1–2 mm) diameter airways of the rabbit (108.3 ± 7.1 mM, n = 5) and monkey (128.5 ± 6.8 mM, n = 3). The measured [Cl−], in excess of 100 mM throughout all airway regions tested in multiple species, is consistent with the isotonic volume hypothesis to describe ASL physiology.

Sign in / Sign up

Export Citation Format

Share Document