Characterization of the in vitro, ex vivo, and in vivo Efficacy of the Antimicrobial Peptide DPK-060 Used for Topical Treatment

AbstractThe in vitro and in vivo characterization of intravitreal injections plays an important role in developing innovative therapy approaches. Using the established vitreous model (VM) and eye movement system (EyeMoS) the distribution of contrast agents with different molecular weight was studied in vitro. The impact of the simulated age-related vitreal liquefaction (VL) on drug distribution in VM was examined either with injection through the gel phase or through the liquid phase. For comparison the distribution was studied ex vivo in the porcine vitreous. The studies were performed in a magnetic resonance (MR) scanner. As expected, with increasing molecular weight the diffusion velocity and the visual distribution of the injected substances decreased. Similar drug distribution was observed in VM and in porcine eye. VL causes enhanced convective flow and faster distribution in VM. Confirming the importance of the injection technique in progress of VL, injection through gelatinous phase caused faster distribution into peripheral regions of the VM than following injection through liquefied phase. VM and MR scanner in combination present a new approach for the in vitro characterization of drug release and distribution of intravitreal dosage forms.

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In vitro , ex vivo and in vivo characterization of PLGA nanoparticles loading pranoprofen for ocular administration

International Journal of Pharmaceutics ◽

10.1016/j.ijpharm.2016.07.055 ◽

2016 ◽

Vol 511 (2) ◽

pp. 719-727 ◽

Cited By ~ 28

Author(s):

Cristina Cañadas ◽

Helen Alvarado ◽

Ana C. Calpena ◽

Amélia M. Silva ◽

Eliana B. Souto ◽

...

Keyword(s):

Ex Vivo ◽

Plga Nanoparticles ◽

In Vivo Characterization

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Erratum to “A first-in-class CDK4 inhibitor demonstrates in vitro, ex-vivo and in vivo efficacy against ovarian cancer” [Gynecologic Oncology 159 (2020) 827–838]

Gynecologic Oncology ◽

10.1016/j.ygyno.2021.07.018 ◽

2021 ◽

Author(s):

Laychiluh Bantie ◽

Solomon Tadesse ◽

Jimma Likisa ◽

Mingfeng Yu ◽

Benjamin Noll ◽

...

Keyword(s):

Ovarian Cancer ◽

Ex Vivo ◽

Gynecologic Oncology ◽

In Vivo Efficacy ◽

Cdk4 Inhibitor

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Formulation Strategies, Characterization, and In Vitro Evaluation of Lecithin-Based Nanoparticles for siRNA Delivery

Journal of Drug Delivery ◽

10.1155/2012/986265 ◽

2012 ◽

Vol 2012 ◽

pp. 1-9 ◽

Cited By ~ 4

Author(s):

Sebastián Ezequiel Pérez ◽

Yamila Gándola ◽

Adriana Mónica Carlucci ◽

Lorena González ◽

Daniel Turyn ◽

...

Keyword(s):

Ex Vivo ◽

Sirna Delivery ◽

Physicochemical Characterization ◽

Human Breast ◽

Efficient Delivery ◽

Interfering Rna ◽

Mcf 7

The aim of the present work was to take advantage of lecithin’s biocompatibility along with its physicochemical properties for the preparation of lecithin-based nanocarriers for small interfering RNA (siRNA) delivery. Water lecithin dispersions were prepared in different conditions, loaded with siRNA at different N/P ratios, and evaluated for loading capacity. The most appropriate ones were then assayed for cytotoxicity and characterized in terms of particle size distribution, zeta potential, and morphology. Results demonstrated that formulations prepared at pH 5.0 and 7.0 were able to load siRNA at broad N/P ratios, and cellular uptake assays showed an efficient delivery of oligos in MCF-7 human breast cancer cells; fluorescent-labeled dsRNA mainly located next to its target, near the nucleus of the cells. No signs of toxicity were observed for broad compositions of lecithin. The physicochemical characterization of the siRNA-loaded dispersions exhibited particles of nanometric sizes and pH-dependant shapes, which make them suitable for ex vivo and in vivo further evaluation.

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Novel 2,4-diaminopyrimidines bearing tetrahydronaphthalenyl moiety against anaplastic lymphoma kinase (ALK): Synthesis, in vitro, ex vivo, and in vivo efficacy studies

Bioorganic & Medicinal Chemistry Letters ◽

10.1016/j.bmcl.2016.02.052 ◽

2016 ◽

Vol 26 (7) ◽

pp. 1720-1725 ◽

Cited By ~ 2

Author(s):

Dawn Song ◽

Minji Lee ◽

Chi Hoon Park ◽

Sunjoo Ahn ◽

Chang Soo Yun ◽

...

Keyword(s):

Anaplastic Lymphoma Kinase ◽

Ex Vivo ◽

In Vivo Efficacy ◽

Anaplastic Lymphoma ◽

Efficacy Studies

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The Antimicrobial Peptide, Bactenecin 5, Supports Cell-Mediated but Not Humoral Immunity in the Context of a Mycobacterial Antigen Vaccine Model

Antibiotics ◽

10.3390/antibiotics9120926 ◽

2020 ◽

Vol 9 (12) ◽

pp. 926

Author(s):

Tulika Munshi ◽

Adam Sparrow ◽

Brendan W. Wren ◽

Rajko Reljic ◽

Samuel J. Willcocks

Keyword(s):

Antimicrobial Peptide ◽

Ex Vivo ◽

Mycobacterial Antigen ◽

Vaccine Adjuvants ◽

Human Tonsil ◽

Mhc I ◽

Surface Marker Expression ◽

Bacillus Subtilis Spore

Bactenecin (Bac) 5 is a bovine antimicrobial peptide (AMP) capable of killing some species of bacteria through the inhibition of protein synthesis. Bac5 and other AMPs have also been shown to have chemotactic properties and can induce inflammatory cytokine expression by innate immune cells. Recently, AMPs have begun to be investigated for their potential use as novel vaccine adjuvants. In the current work, we characterise the functionality of Bac5 in vitro using murine macrophage-like cells, ex vivo using human tonsil tissue and in vivo using a murine model of vaccination. We report the effects of the peptide in isolation and in the context of co-presentation with mycobacterial antigen and whole, inert Bacillus subtilis spore antigens. We find that Bac5 can trigger the release of nitric oxide from murine macrophages and upregulate surface marker expression including CD86, MHC-I and MHC-II, in the absence of additional agonists. When coupled with mycobacterial Ag85 and B. subtilis spores, Bac5 also enhanced IFNγ secretion. We provide evidence that B. subtilis spores, but not the Bac5 peptide, act as strong adjuvants in promoting antigen-specific immunoglobulin production in Ag85B-vaccinated mice. Our findings suggest that Bac5 is an important regulator of the early cell-mediated host immune response.

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Characterization, Stability, and In Vivo Efficacy Studies of Recombinant Human CNTF and Its Permeation into the Neural Retina in Ex Vivo Organotypic Retinal Explant Culture Models

Pharmaceutics ◽

10.3390/pharmaceutics12070611 ◽

2020 ◽

Vol 12 (7) ◽

pp. 611 ◽

Cited By ~ 1

Author(s):

Jaakko Itkonen ◽

Ada Annala ◽

Shirin Tavakoli ◽

Blanca Arango-Gonzalez ◽

Marius Ueffing ◽

...

Keyword(s):

Ex Vivo ◽

Disease Model ◽

Neural Retina ◽

Biologically Active ◽

Target Cells ◽

Therapeutic Effects ◽

Duration Of Action ◽

In Vivo Efficacy

Ciliary neurotrophic factor (CNTF) is one of the most studied neuroprotective agents with acknowledged potential in treating diseases of the posterior eye segment. Although its efficacy and mechanisms of action in the retina have been studied extensively, it is still not comprehensively understood which retinal cells mediate the therapeutic effects of CNTF. As with therapeutic proteins in general, it is poorly elucidated whether exogenous CNTF administered into the vitreous can enter and distribute into the retina and hence reach potentially responsive target cells. Here, we have characterized our purified recombinant human CNTF (rhCNTF), studied the protein’s in vitro bioactivity in a cell-based assay, and evaluated the thermodynamic and oligomeric status of the protein during storage. Biological activity of rhCNTF was further evaluated in vivo in an animal model of retinal degeneration. The retinal penetration and distribution of rhCNTF after 24 h was studied utilizing two ex vivo retina models. Based on our characterization findings, our rhCNTF is correctly folded and biologically active. Moreover, based on initial screening and subsequent follow-up, we identified two buffers in which rhCNTF retains its stability during storage. Whereas rhCNTF did not show photoreceptor preservative effect or improve the function of photoreceptors in vivo, this could possibly be due to the used disease model or the short duration of action with a single intravitreal injection of rhCNTF. On the other hand, the lack of in vivo efficacy was shown to not be due to distribution limitations; permeation into the retina was observed in both retinal explant models as in 24 h rhCNTF penetrated the inner limiting membrane, and being mostly observed in the ganglion cell layer, distributed to different layers of the neural retina. As rhCNTF can reach deeper retinal layers, in general, having direct effects on resident CNTF-responsive target cells is plausible.

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Adult human circulating CD34−Lin−CD45−CD133− cells can differentiate into hematopoietic and endothelial cells

Blood ◽

10.1182/blood-2010-10-316596 ◽

2011 ◽

Vol 118 (8) ◽

pp. 2105-2115 ◽

Cited By ~ 18

Author(s):

Elisa Ciraci ◽

Silvia Della Bella ◽

Ombretta Salvucci ◽

Cristina Rofani ◽

Marta Segarra ◽

...

Keyword(s):

Endothelial Cells ◽

Ex Vivo ◽

Functional Characterization ◽

Adult Life ◽

Hematopoietic Tissue ◽

Developmental Potential ◽

Adult Human

Abstract A precise identification of adult human hemangioblast is still lacking. To identify circulating precursors having the developmental potential of the hemangioblast, we established a new ex vivo long-term culture model supporting the differentiation of both hematopoietic and endothelial cell lineages. We identified from peripheral blood a population lacking the expression of CD34, lineage markers, CD45 and CD133 (CD34−Lin−CD45−CD133− cells), endowed with the ability to differentiate after a 6-week culture into both hematopoietic and endothelial lineages. The bilineage potential of CD34−Lin−CD45−CD133− cells was determined at the single-cell level in vitro and was confirmed by transplantation into NOD/SCID mice. In vivo, CD34−Lin−CD45−CD133− cells showed the ability to reconstitute hematopoietic tissue and to generate functional endothelial cells that contribute to new vessel formation during tumor angiogenesis. Molecular characterization of CD34−Lin−CD45−CD133− cells unveiled a stem cell profile compatible with both hematopoietic and endothelial potentials, characterized by the expression of c-Kit and CXCR4 as well as EphB4, EphB2, and ephrinB2. Further molecular and functional characterization of CD34−Lin−CD45−CD133− cells will help dissect their physiologic role in blood and blood vessel maintenance and repair in adult life.

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