Spray-Congealing and Wet-Sieving as Alternative Processes for Engineering of Inhalation Carrier Particles: Comparison of Surface Properties, Blending and In Vitro Performance

Abstract Purpose Traditionally, α-lactose monohydrate is the carrier of choice in dry powder inhaler (DPI) formulations. Nonetheless, other sugars, such as D-mannitol, have emerged as potential alternatives. Herein, we explored different particle engineering processes to produce D-mannitol carriers for inhaled delivery. Methods Wet-sieving and spray-congealing were employed as innovative techniques to evaluate the impact of engineering on the particle properties of D-mannitol. To that end, the resulting powders were characterized concerning their solid-state, micromeritics and flowability. Afterwards, the engineered carrier particles were blended with inhalable size beclomethasone dipropionate to form low dose (1 wt%) DPI formulations. The in vitro aerosolization performance was evaluated using the NEXThaler®, a reservoir multi-dose device. Results Wet-sieving generated D-mannitol particles with a narrow particle size distribution and spray-congealing free-flowing spherical particles. The more uniform pumice particles with deep voids and clefts of wet-sieved D-mannitol (Pearl300_WS) were beneficial to drug aerosolization, only when used in combination with a ternary agent (10 wt% of ‘Preblend’). When compared to the starting material, the spray-congealed D-mannitol has shown to be promising in terms of the relative increase of the fine particle fraction of the drug (around 100%), when used without the addition of ternary agents. Conclusions The wet-sieving process and the related aerosolization performance are strongly dependent on the topography and structure of the starting material. Spray-congealing, has shown to be a potential process for generating smooth spherical particles of D-mannitol that enhance the in vitro aerosolization performance in binary blends of the carrier with a low drug dose.

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Precise, high-throughput production of multicellular spheroids with a bespoke 3D bioprinter

10.1101/2020.04.06.028548 ◽

2020 ◽

Cited By ~ 1

Author(s):

Robert H. Utama ◽

Lakmali Atapattu ◽

Aidan P. O’Mahony ◽

Christopher M. Fife ◽

Jongho Baek ◽

...

Keyword(s):

Cell Cultures ◽

High Throughput ◽

Cell Number ◽

Drug Dose ◽

3D Bioprinting ◽

Multicellular Spheroids ◽

Drop On Demand ◽

3D Cell Cultures ◽

The Impact

Abstract3D in vitro cancer models are important therapeutic and biological discovery tools, yet formation of multicellular spheroids in a throughput and highly controlled manner to achieve robust and statistically relevant data, remains challenging. Here, we developed an enabling technology consisting of a bespoke drop-on-demand 3D bioprinter capable of high-throughput printing of 96-well plates of spheroids. 3D-multicellular spheroids are embedded inside a tissue-like matrix with precise control over size and cell number. Application of 3D bioprinting for high-throughput drug screening was demonstrated with doxorubicin. Measurements showed that IC50 values were sensitive to spheroid size, embedding and how spheroids conform to the embedding, revealing parameters shaping biological responses in these models. Our study demonstrates the potential of 3D bioprinting as a robust high-throughput platform to screen biological and therapeutic parameters.Significance StatementIn vitro 3D cell cultures serve as more realistic models, compared to 2D cell culture, for understanding diverse biology and for drug discovery. Preparing 3D cell cultures with defined parameters is challenging, with significant failure rates when embedding 3D multicellular spheroids into extracellular mimics. Here, we report a new 3D bioprinter we developed in conjunction with bioinks to allow 3D-multicellular spheroids to be produced in a high-throughput manner. High-throughput production of embedded multicellular spheroids allowed entire drug-dose responses to be performed in 96-well plate format with statistically relevant numbers of data points. We have deconvoluted important parameters in drug responses including the impact of spheroid size and embedding in an extracellular matrix mimic on IC50 values.

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Advanced Microparticulate/Nanoparticulate Respirable Dry Powders of a Selective RhoA/Rho Kinase (Rock) Inhibitor for Targeted Pulmonary Inhalation Aerosol Delivery

Pharmaceutics ◽

10.3390/pharmaceutics13122188 ◽

2021 ◽

Vol 13 (12) ◽

pp. 2188

Author(s):

Priya Muralidharan ◽

Don Hayes ◽

Jeffrey R. Fineman ◽

Stephen M. Black ◽

Heidi M. Mansour

Keyword(s):

Pulmonary Hypertension ◽

Physicochemical Characterization ◽

Rho Kinase ◽

Drug Dose ◽

Particle Engineering ◽

Rock Inhibitor ◽

Medical Needs ◽

Human Lung Cells ◽

Aerosol Dispersion

Pulmonary hypertension (PH) is a progressive disease that eventually leads to heart failure and potentially death for some patients. There are many unique advantages to treating pulmonary diseases directly and non-invasively by inhalation aerosols and dry powder inhalers (DPIs) possess additional unique advantages. There continues to be significant unmet medical needs in the effective treatment of PH that target the underlying mechanisms. To date, there is no FDA-approved DPI indicated for the treatment of PH. Fasudil is a novel RhoA/Rho kinase (ROCK) inhibitor that has shown great potential in effectively treating pulmonary hypertension. This systematic study is the first to report on the design and development of DPI formulations comprised of respirable nanoparticles/microparticles using particle engineering design by advanced spray drying. In addition, comprehensive physicochemical characterization, in vitro aerosol aerosol dispersion performance with different types of human DPI devices, in vitro cell-drug dose response cell viability of different human respiratory cells from distinct lung regions, and in vitro transepithelial electrical resistance (TEER) as air-interface culture (AIC) demonstrated that these innovative DPI fasudil formulations are safe on human lung cells and have high aerosol dispersion performance properties.

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Wear of different materials for total hip replacement under adverse stop-dwell-start in vitro wear simulation conditions

Proceedings of the Institution of Mechanical Engineers Part H Journal of Engineering in Medicine ◽

10.1177/0954411918813385 ◽

2018 ◽

Vol 232 (12) ◽

pp. 1261-1270 ◽

Cited By ~ 1

Author(s):

Megan Hadley ◽

Catherine Hardaker ◽

Graham Isaac ◽

John Fisher

Keyword(s):

Total Hip Arthroplasty ◽

Hip Arthroplasty ◽

Activity Cycle ◽

Relative Increase ◽

Total Hip ◽

Metal On Metal ◽

Metal Metal ◽

The Impact ◽

Ceramic On Ceramic

Hip simulation is a common technique for pre-clinical evaluation of wear performance of total hip arthroplasty. Standard techniques replicate kinematics of walking patterns of a typical patient. Attention has focussed in developing simulations of other typical patient daily activities to improve accuracy of wear predictions. A method for simulating stop-dwell-start motion during patient walking and the effect on 36-mm metal-on-metal total hip arthroplasty was previously presented by the authors. This study sought to extend the previous work to look at the effect of these conditions on ceramic-on-ceramic, metal-on-polyethylene and ceramic-on-polyethylene bearings. Two stop-dwell-start protocols were used: one reproducing average patient movement patterns and one examining more severe conditions. For all materials tested, no significant increase in wear was observed under average stop-dwell-start conditions, suggesting the bearing types tested are robust to this type of activity. A significant increase in wear was observed for metal-on-metal, metal-on-polyethylene and ceramic-on-polyethylene bearings under severe stop-dwell-start conditions, this was attributed to depletion of lubricant in the bearing during the dwell period. A greater relative increase in wear was observed for metal-on-metal bearings compared with metal-on-polyethylene and ceramic-on-polyethylene bearings. This may be explained by the contributions of the different lubrication mechanisms in each bearing type. Wear of ceramic-on-ceramic was very low in all tests, suggesting normal measurement variation was masking any effect of the adverse conditions. It was not possible to determine any effect of the different activities. These results emphasise the importance of exploring adverse patient activity simulations. The increase in wear rate associated with an adverse activity such as seen in stop-dwell-start motion, has to be considered in the context of the frequency of the adverse activity cycle relative to other activities such as standard continuous walking, to determine the impact on the total wear in a given time period.

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Evaluation of the Impact of Excipients on Albendazole Concentrations in Upper Small Intestine Using an In Vitro Biorelevant Gastrointestinal Transfer (BioGIT) System

10.26226/morressier.57d6b2bbd462b8028d88e13f ◽

2016 ◽

Author(s):

Maria Vertzoni

Keyword(s):

Small Intestine ◽

The Impact

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Development and Characterization Colchicine-Loaded PEGylated Gelatin Nanoparticles for Targeted Delivery to Tumor

International Journal of Pharmaceutical Sciences and Nanotechnology ◽

10.37285/ijpsn.2013.6.2.8 ◽

2013 ◽

Vol 6 (2) ◽

pp. 2058-2063

Author(s):

G D Chandrethiya ◽

P K Shelat ◽

M N Zaveri

Keyword(s):

Drug Delivery ◽

Targeted Delivery ◽

High Performance ◽

Entrapment Efficiency ◽

Stability Study ◽

Spherical Particles ◽

Precipitation Method ◽

Antitumoral Activity ◽

Gelatin Nanoparticles

PEGylated gelatin nanoparticles loaded with colchicine were prepared by ethanol precipitation method. Poly-(ethylene glycol)-5000-monomethylether (MPEG 5000), a hydrophilic polymer, was used to pegylate gelatin. Gluteraldehyde was used as cross-linking agent. To obtain a high quality product, major formulation parameters were optimized. Spherical particles with mean particles of 193 nm were measured by a Malvern particle size analyzer. Entrapment efficiency was found to be 71.7 ± 1.4% and determined with reverse phase high performance liquid charomatography (RP-HPLC). The in vitro drug release study was performed by dialysis bag method for a period of 168 hours. Lyophilizaton study showed sucrose at lower concentrations proved the best cryoprotectant for this formulation. Stability study revealed that lyophilized nanoparticles were equally effective (p < 0.05) after one year of storage at 2-8°C with ambient humidity. In vitro antitumoral activity was accessed using the MCF-7 cell line by MTT assay. The IC50 value was found to be 0.034 μg/ml for the prepared formulation. The results indicate that PEGylated gelatin nanoparticles could be utilized as a potential drug delivery for targeted drug delivery of tumors.

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The Predominant microRNAs in β-cell Clusters for Insulin Regulation and Diabetic Control

Current Drug Targets ◽

10.2174/1389450121666191230145848 ◽

2020 ◽

Vol 21 (7) ◽

pp. 722-734

Author(s):

Adele Soltani ◽

Arefeh Jafarian ◽

Abdolamir Allameh

Keyword(s):

Mirna Expression ◽

Expression Profiles ◽

Expression Patterns ◽

Diabetic Control ◽

In Vitro Proliferation ◽

Cell Functions ◽

Mirna Expression Profiles ◽

Multiple Processes ◽

The Impact

micro (mi)-RNAs are vital regulators of multiple processes including insulin signaling pathways and glucose metabolism. Pancreatic β-cells function is dependent on some miRNAs and their target mRNA, which together form a complex regulative network. Several miRNAs are known to be directly involved in β-cells functions such as insulin expression and secretion. These small RNAs may also play significant roles in the fate of β-cells such as proliferation, differentiation, survival and apoptosis. Among the miRNAs, miR-7, miR-9, miR-375, miR-130 and miR-124 are of particular interest due to being highly expressed in these cells. Under diabetic conditions, although no specific miRNA profile has been noticed, the expression of some miRNAs and their target mRNAs are altered by posttranscriptional mechanisms, exerting diverse signs in the pathobiology of various diabetic complications. The aim of this review article is to discuss miRNAs involved in the process of stem cells differentiation into β-cells, resulting in enhanced β-cell functions with respect to diabetic disorders. This paper will also look into the impact of miRNA expression patterns on in vitro proliferation and differentiation of β-cells. The efficacy of the computational genomics and biochemical analysis to link the changes in miRNA expression profiles of stem cell-derived β-cells to therapeutically relevant outputs will be discussed as well.

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The Impact of Antioxidants from the Diet on Breast Cancer Cells Monitored by Raman Microspectroscopy

Letters in Drug Design & Discovery ◽

10.2174/1570180815666180502120804 ◽

2018 ◽

Vol 16 (2) ◽

pp. 127-137

Author(s):

Paula Sofia Coutinho Medeiros ◽

Ana Lúcia Marques Batista de Carvalho ◽

Cristina Ruano ◽

Juan Carlos Otero ◽

Maria Paula Matos Marques

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Cell Line ◽

Breast Cancer Cells ◽

Triple Negative ◽

Breast Adenocarcinoma ◽

Coumaric Acid ◽

Human Breast ◽

The Impact

Background: The impact of the ubiquitous dietary phenolic compound p-coumaric acid on human breast cancer cells was assessed, through a multidisciplinary approach: Combined biological assays for cytotoxicity evaluation and biochemical profiling by Raman microspectroscopic analysis in cells. </P><P> Methods: Para-coumaric acid was shown to exert in vitro chemoprotective and antitumor activities, depending on the concentration and cell line probed: a significant anti-invasive ability was detected for the triple-negative MDA-MB-231 cells, while a high pro-oxidant effect was found for the estrogen- dependent MCF-7 cells. A striking cell selectivity was obtained, with a more noticeable outcome on the triple-negative MDA-MB-231 cell line. Results: The main impact on the cellular biochemical profile was verified to be on proteins and lipids, thus justifying the compound´s anti-invasive effect and chemoprotective ability. Conclusion: p-Coumaric acid was thus shown to be a promising chemoprotective/chemotherapeutic agent, particularly against the low prognosis triple-negative human breast adenocarcinoma.

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VDAC1 Mediated Anticancer Activity of Gallic Acid in Human Lung Adenocarcinoma A549 Cells

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520617666170912115441 ◽

2018 ◽

Vol 18 (2) ◽

pp. 255-262 ◽

Cited By ~ 5

Author(s):

Aikebaier Maimaiti ◽

Amier Aili ◽

Hureshitanmu Kuerban ◽

Xuejun Li

Keyword(s):

Western Blot ◽

Cell Viability ◽

Gallic Acid ◽

Molecular Mechanisms ◽

A549 Cells ◽

Dependent Manner ◽

Lung Carcinoma Cells ◽

Induced Apoptosis ◽

The Impact

Aims: Gallic acid (GA) is generally distributed in a variety of plants and foods, and possesses cell growth-inhibiting activities in cancer cell lines. In the present study, the impact of GA on cell viability, apoptosis induction and possible molecular mechanisms in cultured A549 lung carcinoma cells was investigated. Methods: In vitro experiments showed that treating A549 cells with various concentrations of GA inhibited cell viability and induced apoptosis in a dose-dependent manner. In order to understand the mechanism by which GA inhibits cell viability, comparative proteomic analysis was applied. The changed proteins were identified by Western blot and siRNA methods. Results: Two-dimensional electrophoresis revealed changes that occurred to the cells when treated with or without GA. Four up-regulated protein spots were clearly identified as malate dehydrogenase (MDH), voltagedependent, anion-selective channel protein 1(VDAC1), calreticulin (CRT) and brain acid soluble protein 1(BASP1). VDAC1 in A549 cells was reconfirmed by western blot. Transfection with VDAC1 siRNA significantly increased cell viability after the treatment of GA. Further investigation showed that GA down regulated PI3K/Akt signaling pathways. These data strongly suggest that up-regulation of VDAC1 by GA may play an important role in GA-induced, inhibitory effects on A549 cell viability.

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Effects of Transport Medium Composition on in vitro Drug Permeation across Excised Pig Intestinal Tissue

Drug Delivery Letters ◽

10.2174/2210303110999201005214114 ◽

2020 ◽

Vol 10 ◽

Author(s):

Bianca Peterson ◽

Henrico Heystek ◽

Josias H. Hamman ◽

Johan D. Steyn

Keyword(s):

Medium Composition ◽

Screening Tests ◽

Intestinal Tissue ◽

Predictive Values ◽

Transport Medium ◽

Intestinal Fluids ◽

Lower Permeability ◽

The Impact ◽

Simulated Intestinal Fluids

Background:: Knowledge of the permeation characteristics of new chemical entities across biological membranes is essential to drug research and development. Transport medium composition may affect the absorption of compounds during in vitro drug transport testing. To preserve the predictive values of screening tests, the possible influence of transport media on the solubility of model drugs, and on the activities of tight junctions and efflux transporter proteins (e.g. P-glycoprotein) must be known. Objective:: The aim of this study was to compare the impact of different transport media on the bi-directional transport of standard compounds, selected from the four classes of the Biopharmaceutical Classification System (BCS), across excised pig intestinal tissue. Methods:: The Sweetana-Grass diffusion apparatus was used for the transport studies. Krebs-Ringer bicarbonate (KRB) buffer and simulated intestinal fluids in the fed (FeSSIF) and fasted (FaSSIF) states were used as the three transport media, while the chosen compounds were abacavir (BCS class 1), dapsone (BCS class 2), lamivudine (BCS class 3) and furosemide (BCS class 4). Results:: Abacavir exhibited lower permeability in both the simulated intestinal fluids than in the KRB buffer. Dapsone showed similar permeability in all media. Lamivudine exhibited lower permeability in FaSSIF than in the other two media. Furosemide exhibited improved transport with pronounced efflux in FaSSIF. Conclusion:: Different permeation behaviors were observed for the selected drugs in the respective media, which may have resulted from their different physico-chemical properties, as well as from the effects that dissimilar transport media components had on excised pig intestinal tissue.

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