Testing of aerosolized ciprofloxacin nanocarriers on cystic fibrosis airway cells infected with P. aeruginosa biofilms

AbstractThe major pathogen found in the lungs of adult cystic fibrosis (CF) patients is Pseudomonas aeruginosa, which builds antibiotic-resistant biofilms. Pulmonary delivery of antibiotics by inhalation has already been proved advantageous in the clinic, but the development of novel anti-infective aerosol medicines is complex and could benefit from adequate in vitro test systems. This work describes the first in vitro model of human bronchial epithelial cells cultivated at the air–liquid interface (ALI) and infected with P. aeruginosa biofilm and its application to demonstrate the safety and efficacy of aerosolized anti-infective nanocarriers. Such a model may facilitate the translation of novel therapeutic modalities into the clinic, reducing animal experiments and the associated problems of species differences. A preformed biofilm of P. aeruginosa PAO1 was transferred to filter-grown monolayers of the human CF cell line (CFBE41o-) at ALI and additionally supplemented with human tracheobronchial mucus. This experimental protocol provides an appropriate time window to deposit aerosolized ciprofloxacin-loaded nanocarriers at the ALI. When applied 1 h post-infection, the nanocarriers eradicated all planktonic bacteria and reduced the biofilm fraction of the pathogen by log 6, while CFBE41o- viability and barrier properties were maintained. The here described complex in vitro model approach may open new avenues for preclinical safety and efficacy testing of aerosol medicines against P. aeruginosa lung infection. Graphical abstract

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Essential genome ofPseudomonas aeruginosain cystic fibrosis sputum

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1419677112 ◽

2015 ◽

Vol 112 (13) ◽

pp. 4110-4115 ◽

Cited By ~ 164

Author(s):

Keith H. Turner ◽

Aimee K. Wessel ◽

Gregory C. Palmer ◽

Justine L. Murray ◽

Marvin Whiteley

Keyword(s):

Cystic Fibrosis ◽

In Vitro Model ◽

Core Genome ◽

Growth Conditions ◽

Sequencing Data ◽

Orthologous Genes ◽

Statistical Precision ◽

Simulation Based ◽

Vitro Model

Defining the essential genome of bacterial pathogens is central to developing an understanding of the biological processes controlling disease. This has proven elusive forPseudomonas aeruginosaduring chronic infection of the cystic fibrosis (CF) lung. In this paper, using a Monte Carlo simulation-based method to analyze high-throughput transposon sequencing data, we establish theP. aeruginosaessential genome with statistical precision in laboratory media and CF sputum. Reconstruction of the global requirements for growth in CF sputum compared with defined growth conditions shows that the latter requires several cofactors including biotin, riboflavin, and pantothenate. Comparison ofP. aeruginosastrains PAO1 and PA14 demonstrates that essential genes are primarily restricted to the core genome; however, some orthologous genes in these strains exhibit differential essentiality. These results indicate that genes with similar molecular functions may have distinct genetic roles in differentP. aeruginosastrains during growth in CF sputum. We also show that growth in a defined growth medium developed to mimic CF sputum yielded virtually identical fitness requirements to CF sputum, providing support for this medium as a relevant in vitro model for CF microbiology studies.

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Acute drug transfer and particle release of drug coated balloons within a porcine in-vitro model

Current Directions in Biomedical Engineering ◽

10.1515/cdbme-2017-0098 ◽

2017 ◽

Vol 3 (2) ◽

pp. 465-468

Author(s):

Christoph Brandt-Wunderlich ◽

Lucas Almstädt ◽

Sebastian Kaule ◽

Thomas Reske ◽

Wolfram Schmidt ◽

...

Keyword(s):

In Vitro Model ◽

Test Methods ◽

In Vivo Studies ◽

In Vitro Test ◽

Drug Transfer ◽

Particle Release ◽

Artery Disease ◽

Vitro Model

AbstractDrug coated balloons (DCB) are used in the therapy of coronary as well as peripheral artery disease. The success of drug transfer to the vessel wall depends on the excipient used in combination with paclitaxel as antiproliferative drug. Although in-vivo studies show very good results with this technology, there is a lack of in-vitro test methods for characterization of various DCB available on the market. This study describes a method to gain information about the drug transfer and the particle release of three different DCB based on cetylpyridinium salycate (Cetpyrsal), hyaluronic acid and iopromide within a porcine in-vitro model. The Cetpyrsal-based DCB showed promising results with the highest drug transfer while producing the lowest number of particles.

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The Coexistence of Cystic Fibrosis and Celiac Disease

PEDIATRICS ◽

10.1542/peds.57.5.715 ◽

1976 ◽

Vol 57 (5) ◽

pp. 715-721

Author(s):

Aubrey J. Katz ◽

Z. Myron Falchuk ◽

Harry Shwachman

Keyword(s):

Cystic Fibrosis ◽

Celiac Disease ◽

Organ Culture ◽

Intestinal Mucosa ◽

In Vitro Model ◽

Duodenal Mucosa ◽

Common Type ◽

The Other ◽

Vitro Model

Two patients with cystic fibrosis (CF) who subsequently developed celiac disease (CD) are described. Since organ culture of intestinal mucosa has been used to establish an in vitro model for the study of CD, we utilized this opportunity to determine whether duodenal mucosa obtained from each of these two patients and their immediate families differed in its organ culture behavior from mucosa obtained from patients with CD alone. Additionally, as specific HL-A types are associated with CD, we used HL-A typing to determine whether the two patients with CF-CD differed genetically from patients with CD alone. One of our patients was HL-A8, the most common type associated with CD; the other was HL-A12, as are many of the non-HL-A8 celiac patients. The response in organ culture of the mucosa of these two patients was the same as the response in organ culture of the mucosa from patients with CD alone. These and other data suggest that CD occurring in patients with CF is no different than CD occurring alone.

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Therapeutic time window for the effects of erythropoietin on astrogliosis and neurite outgrowth in an in vitro model of spinal cord injury

Medicine ◽

10.1097/md.0000000000009913 ◽

2018 ◽

Vol 97 (9) ◽

pp. e9913 ◽

Cited By ~ 4

Author(s):

Hea Nam Hong ◽

Ju Hee Shim ◽

You Jin Won ◽

Jong Yoon Yoo ◽

Chang Ho Hwang

Keyword(s):

Spinal Cord ◽

Spinal Cord Injury ◽

Neurite Outgrowth ◽

In Vitro Model ◽

Time Window ◽

Cord Injury ◽

Vitro Model ◽

Therapeutic Time Window

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Critical Evaluation of an in Vitro Model for Evaluation of Platelet Reactivity of Biomaterials

MRS Proceedings ◽

10.1557/proc-110-325 ◽

1987 ◽

Vol 110 ◽

Author(s):

Raymond Connolly ◽

Norman Shoenfeld ◽

Karen Ramberg ◽

Allan D. Callow

Keyword(s):

In Vitro Model ◽

Platelet Reactivity ◽

Platelet Rich Plasma ◽

Critical Evaluation ◽

The Body ◽

Test Material ◽

In Vitro Test ◽

Vitro Model

AbstractAn in vitro model for measuring platelet reactivity to a variety of biomaterial candidates for vascular grafts is described. A model consisting of a standard area of test material exposed to freshly labeled In platelets in plasma was evaluated. The platelets were isolated from ACD anticoagulated blood and resuspended in ACD plasma. It has been previously demonstrated that platelets so treated circulate in the body and will deposit on biomaterials exposed to the blood in vivo. The in vitro test consisted of an incubation of the platelets and materials at 37°C for one hour. At the end of the incubation, the platelet rich plasma was removed and the materials washed and removed for gamma counting. Platelet reactivity was normalized as a percentage of the counts on the material to counts in an aliquot of the platelet-plasma incubation media. The maximum uptake of platelets occurred within one hour. Platelets from three species, human, baboon, and dog were tested. Platelet uptake by Dacron and PTFE were in the range of 30–40% and 1–5% respectively. This is in accord with the known reactivity of these two vascular graft materials in vivo.A second series of studies were conducted with physically and pharmacologically inactivated platelets and inert particles. Those studies suggest that the initial results do not represent a biologic event but may reflect the porosity of the materials. This emphasizes the necessity of adequately defining an in vitro model against known in vivo activity.

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