RESUMO
Oral antibiotic treatment is often applied in animal studies in order to allow establishment of an introduced antibiotic-resistant bacterium in the gut. Here, we compared the application of streptomycin dosed orally in microcontainers to dosage through drinking water. The selective effect on a resistant bacterial strain, as well as the effects on fecal, luminal, and mucosal microbiota composition, were investigated. Three groups of rats (n = 10 per group) were orally dosed with microcontainers daily for 3 days. One of these groups (STR-M) received streptomycin-loaded microcontainers designed for release in the distal ileum, while the other two groups (controls [CTR] and STR-W) received empty microcontainers. The STR-W group was additionally dosed with streptomycin through the drinking water. A streptomycin-resistant Escherichia coli strain was orally inoculated into all animals. Three days after inoculation, the resistant E. coli was found only in the cecum and colon of animals receiving streptomycin in microcontainers but in all intestinal compartments of animals receiving streptomycin in the drinking water. 16S rRNA amplicon sequencing revealed significant changes in the fecal microbiota of both groups of streptomycin-treated animals. Investigation of the inner colonic mucus layer by confocal laser scanning microscopy and laser capture microdissection revealed no significant effect of streptomycin treatment on the mucus-inhabiting microbiota or on E. coli encroachment into the inner mucus. Streptomycin-loaded microcontainers thus enhanced proliferation of an introduced streptomycin-resistant E. coli in the cecum and colon without affecting the small intestine environment. While improvements of the drug delivery system are needed to facilitate optimal local concentration and release of streptomycin, the application of microcontainers provides new prospects for antibiotic treatment. IMPORTANCE Delivery of antibiotics in microcontainer devices designed for release at specific sites of the gut represents a novel approach which might reduce the amount of antibiotic needed to obtain a local selective effect. We propose that the application of microcontainers may have the potential to open novel opportunities for antibiotic treatment of humans and animals with fewer side effects on nontarget bacterial populations. In the current study, we therefore elucidated the effects of streptomycin, delivered in microcontainers coated with pH-sensitive lids, on the selective effect on a resistant bacterium, as well as on the surrounding intestinal microbiota in rats.
Assuntos
Água Potável , Estreptomicina , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bactérias/genética , Colo , Escherichia coli/genética , Humanos , Mucosa Intestinal/microbiologia , RNA Ribossômico 16S , Ratos , Estreptomicina/farmacologiaRESUMO
With the growing popularity and application of microfabricated devices in oral drug delivery (ODD), masking technologies for drug loading and surface modification become highly relevant. Considering the speed of design and fabrication processes and the necessity for continuous alterations of e.g. the shape and sizes of the devices during the optimization process, there is a need for adaptable, precise and low-cost masking techniques. Here, a novel method is presented for masking ODD microdevices, namely microcontainers, using the physical characteristics of polydimethylsiloxane (PDMS). When compared to a rigid microfabricated shadow mask, used for filling drugs in microcontainers, the PDMS masking technique allows more facile and precise loading of higher quantities of an active compound, without the need of alignment. The method provides flexibility and is adjustable to devices fabricated from different materials with various geometries, topologies and dimensions. This user-friendly flexible masking method overcomes the limitations of other masking techniques and is certainly not limited to ODD and is recommended for a wide range of microdevices.
Assuntos
Dimetilpolisiloxanos/química , Sistemas de Liberação de Medicamentos/instrumentação , Dispositivos Lab-On-A-Chip , Fenômenos Mecânicos , Administração Oral , Desenho de EquipamentoRESUMO
The distinction between surface and bulk crystallization of amorphous pharmaceuticals, as well as the importance of surface crystallization for pharmaceutical performance, is becoming increasingly evident. An emerging strategy in stabilizing the amorphous drug form is to utilize thin coatings at the surface. While the physical stability of systems coated with pharmaceutical polymers has recently been studied, the effect on dissolution performance as a function of storage time, as a further necessary step toward the success of these formulations, has not been previously studied. Furthermore, the effect of coating thickness has not been elucidated. This study investigated the effect of these polymer-coating parameters on the interplay between amorphous surface crystallization and drug dissolution for the first time. The study utilized simple tablet-like coated dosage forms, comprising a continuous amorphous drug core and thin polymer coating (hundreds of nanometers to a micrometer thick). Monitoring included analysis of both the solid-state of the model drug (with SEM, XRD, and ATR FTIR spectroscopy) and dissolution performance (and associated morphology and solid-state changes) after different storage times. Stabilization of the amorphous form (dependent on the coating thickness) and maintenance of early-stage intrinsic dissolution rates characteristic for the unaged amorphous drug were achieved. However, dissolution in the latter stages was likely inhibited by the presence of a polymer at the surface. Overall, this study introduced a versatile coated system for studying the dissolution of thin-coated amorphous dosage forms suitable for different drugs and coating agents. It demonstrated the importance of multiple factors that need to be taken into consideration when aiming to achieve both physical stability and improved release during the shelf life of amorphous formulations.
Assuntos
Composição de Medicamentos/métodos , Preparações Farmacêuticas/química , Polímeros/química , Química Farmacêutica/métodos , Cristalização/métodos , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Solubilidade , Propriedades de Superfície , Comprimidos/químicaRESUMO
Cell or tissue stretching and strain are present in any in vivo environment, but is difficult to reproduce in vitro. Here, we describe a simple method for casting a thin (about 500 µm) and soft (about 0.3 kPa) hydrogel of gelatin and a method for characterizing the mechanical properties of the hydrogel simply by changing pressure with a water column. The gelatin is crosslinked with mTransglutaminase and the area of the resulting hydrogel can be increased up 13-fold by increasing the radial water pressure. This is far beyond physiological stretches observed in vivo. Actuating the hydrogel with a radial force achieves both information about stiffness, stretchability, and contractability, which are relevant properties for tissue engineering purposes. Cells could be stretched and contracted using the gelatin membrane. Gelatin is a commonly used polymer for hydrogels in tissue engineering, and the discovered reversible stretching is particularly interesting for organ modeling applications.
Assuntos
Gelatina/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Polímeros/química , Engenharia Tecidual , Gelatina/síntese química , Hidrogel de Polietilenoglicol-Dimetacrilato/síntese química , Fenômenos Mecânicos , Membranas/química , Polímeros/síntese química , Transglutaminases/química , Água/químicaRESUMO
The cell-penetrating peptide (CPP) penetratin has demonstrated potential as a carrier for transepithelial delivery of cargo peptides, such as the therapeutically relevant part of parathyroid hormone, i.e., PTH(1-34). The purpose of the present study was to elucidate the relevance of pH for PTH(1-34)-penetratin conjugates and coadministered penetratin with PTH(1-34) regarding transepithelial permeation of PTH(1-34) and cellular effects. Transepithelial permeation was assessed using monolayers of the Caco-2 cell culture model, and effects on Caco-2 cellular viability kinetics were evaluated by using the Real-Time-GLO assay as well as by microscopy following Tryphan blue staining. Morphological Caco-2 cell changes were studied exploiting the impedance-based xCELLigence system as well as optically using the oCelloscope setup. Finally, the effect of pH on the folding propensity of the PTH(1-34)-penetratin conjugate and its ability to disrupt lipid membranes were assessed by circular dichroism (CD) spectroscopy and the calcein release assay, respectively. The transepithelial PTH(1-34) permeation was not pH-dependent when applying the coadministration approach. However, by applying the conjugation approach, the PTH(1-34) permeation was significantly enhanced by lowering the pH from 7.4 to 5 but also associated with a compromised barrier and a lowering of the cellular viability. The negative effects on the cellular viability following cellular incubation with the PTH(1-34)-penetratin conjugate were moreover confirmed during real-time monitoring of the Caco-2 cell viability as well as by enhanced Tryphan blue uptake. In addition, morphological changes were primarily observed for cells incubated with the PTH(1-34)-penetratin conjugate at pH 5, which was moreover demonstrated to have an enhanced membrane permeating effect following lowering of the pH from 7.4 to 5. The latter observation was, however, not a result of better secondary folding propensity at pH 5 when compared to pH 7.4.
Assuntos
Proteínas de Transporte/química , Nanoconjugados/química , Hormônio Paratireóideo/química , Hormônio Paratireóideo/farmacocinética , Sequência de Aminoácidos , Células CACO-2 , Proteínas de Transporte/farmacocinética , Permeabilidade da Membrana Celular , Sobrevivência Celular , Peptídeos Penetradores de Células , Epitélio/metabolismo , Humanos , Concentração de Íons de Hidrogênio , PermeabilidadeRESUMO
A photonic crystal slab (PCS) sensor is a universal refractive index sensor with possibilities and performance very similar to surface plasmon resonance (SPR), which represents the gold standard of biosensing. Cheap PCS sensors can be made vacuum-free entirely out of polymers, but come with additional challenges, besides those relating to temperature-variations, which must be considered in any refractive index based method: The polymeric waveguide core was found to swell by â¼0.3% as water absorbed into the waveguide core over â¼1.5 h. This was investigated by monitoring the wavelength of resonant reflection during absorption, by monitoring the release of water using ellipsometry, and by rigorous coupled-wave analysis (RCWA). The approach presented here enables monitoring of water uptake and thermal fluctuations, for drift-free, high-performance operation of a polymeric PCS sensor.
RESUMO
Chitosan particles loaded with the antigen ovalbumin (OVA) and the adjuvant Quil-A were produced by electrospray, using mixtures of water/ethanol/acetic acid as a solvent. Three different chitosans designed as HMC+70, HMC+85, and HMC+90 (called as 705010, 855010, and 905010) were tested and its efficacy to be used in oral vaccine delivery applications was investigated. The morphology, size, and zeta potential of the produced particles were investigated, together with the encapsulation efficiency and release of OVA from the three chitosan formulations. Moreover, the mucoadhesion and cytotoxicity of the chitosan microparticles was examined. All the three formulations with OVA and Quil-A were in the micrometer size range and had a positive zeta potential between 46 and 75 mV. Furthermore, all the three formulations displayed encapsulation efficiencies above 80% and the release of OVA over a period of 80 h was observed to be between 38 and 47%. None of the developed formulations exhibited high mucoadhesive properties, either cytotoxicity. The formulation prepared with HMC+70, OVA, and Quil-A had the highest stability within 2 h in buffer solution, as measured by dynamic light scattering. The electrosprayed formulation consisting of HMC+70 with OVA and Quil-A showed to be the most promising as an oral vaccine system.
Assuntos
Química Farmacêutica/métodos , Quitosana/síntese química , Sistemas de Liberação de Medicamentos/métodos , Microesferas , Tamanho da Partícula , Vacinas/síntese química , Administração Oral , Animais , Linhagem Celular , Galinhas , Quitosana/administração & dosagem , Composição de Medicamentos , Humanos , Ovalbumina/administração & dosagem , Ovalbumina/síntese química , Saponinas de Quilaia/administração & dosagem , Saponinas de Quilaia/síntese química , Vacinas/administração & dosagemRESUMO
Many drug candidates today have a low aqueous solubility and, hence, may show a low oral bioavailability, presenting a major formulation and drug delivery challenge. One way to increase the bioavailability of these drugs is to use a supersaturating drug delivery strategy. The aim of this study was to develop a video-microscopic method, to evaluate the effect of a precipitation inhibitor on supersaturated solutions of the poorly soluble drug tadalafil, using a novel video-microscopic small scale setup. Based on preliminary studies, a degree of supersaturation of 29 was chosen for the supersaturation studies with tadalafil in FaSSIF. Different amounts of hydroxypropyl methyl cellulose (HPMC) were predissolved in FaSSIF to give four different concentrations, and the supersaturated system was then created using a solvent shift method. Precipitation of tadalafil from the supersaturated solutions was monitored by video-microscopy as a function of time. Single-particle analysis was possible using commercially available software; however, to investigate the entire population of precipitating particles (i.e., their number and area covered in the field of view), an image analysis algorithm was developed (multiparticle analysis). The induction time for precipitation of tadalafil in FaSSIF was significantly prolonged by adding 0.01% (w/v) HPMC to FaSSIF, and the maximum inhibition was reached at 0.1% (w/v) HPMC, after which additional HPMC did not further increase the induction time. The single-particle and multiparticle analyses yielded the same ranking of the HPMC concentrations, regarding the inhibitory effect on precipitation. The developed small scale method to assess the effect of precipitation inhibitors can speed up the process of choosing the right precipitation inhibitor and the concentration to be used.
Assuntos
Precipitação Química/efeitos dos fármacos , Excipientes/farmacologia , Derivados da Hipromelose/farmacologia , Tadalafila/farmacocinética , Algoritmos , Disponibilidade Biológica , Química Farmacêutica , Desenho de Fármacos , Excipientes/química , Derivados da Hipromelose/química , Microscopia de Vídeo/métodos , Modelos Químicos , Solubilidade , Solventes/química , Tadalafila/química , Água/químicaRESUMO
Microwells fabricated from poly-L-lactic acid (PLLA) were evaluated for their application as an oral drug delivery system using the amorphous sodium salt of furosemide (ASSF) as a model drug. Hot embossing of PLLA resulted in fabrication of microwells with an inner diameter of 240 µm and a height of 100 µm. The microwells were filled with ASSF using a modified screen printing technique, followed by coating of the microwell cavities with a gastro-resistant lid of Eudragit® L100. The release behavior of ASSF from the coated microwells was investigated using a µ-Diss profiler and a UV imaging system, and under conditions simulating the changing environment of the gastrointestinal tract. Biorelevant gastric medium (pH 1.6) was employed, after which a change to biorelevant intestinal release medium (pH 6.5) was carried out. Both µ-Diss profiler and UV imaging release experiments showed that sealing of microwell cavities with an Eudragit® layer prevented drug release in biorelevant gastric medium. An immediate release of the ASSF from coated microwells was observed in the intestinal medium. This pH-triggered release behavior demonstrates the future potential of PLLA microwells as a site-specific oral drug delivery system.
Assuntos
Implantes Absorvíveis , Implantes de Medicamento/síntese química , Furosemida/química , Suco Gástrico/química , Concentração de Íons de Hidrogênio , Ácido Láctico/química , Polímeros/química , Administração Oral , Cápsulas , Difusão , Implantes de Medicamento/administração & dosagem , Furosemida/administração & dosagem , Humanos , Teste de Materiais , PoliésteresRESUMO
Oral vaccination has in the recent years gained a lot of attraction, mainly due to optimized patient compliance and logistics. However, the development of oral vaccines, especially oral subunit vaccines is challenging. Micro technology can be utilized to overcome some of these challenges, by facilitating protection and effective delivery of the vaccine components in the gastrointestinal tract (GI tract). One such technology is Microcontainers (MCs), which can be realized to be mucoadhesive and to target specific regions of the GI tract via oral delivery. Here, we test MCs, for oral delivery of the C. trachomatis vaccine candidate CTH522, in combination with effective mucosal adjuvants. The adjuvants alpha- galactosylceramide (α-GalCer), C-di-GMP and cholera toxin B were compared in vivo, to identify the most prominent adjuvant for formulation with CTH522. Formulations were administered both purely oral and as boosters following a subcutaneous (s.c.) prime with CTH522 in combination with the CAF®01 adjuvant. CTH522 formulated with α-GalCer showed to be the most efficient combination for the oral vaccine, based on the immunological analysis. Lyophilized formulation of CTH522 and α-GalCer was loaded into MCs and these were subsequently coated with Eudragit L100-55 and evaluated in vivo in mice for the ability of MCs to mediate intestinal vaccine delivery and increase immunogenicity of the vaccine. Mice receiving oral prime and boosters did show a significantly enhanced mucosal immune responses compared to naive mice. This indicates the MCs are indeed capable of delivering the vaccine formulation intact and able to stimulate the immune cells. Mice orally boosted with MCs following a s.c. prime with CAF01, demonstrated improved systemic and local Th17 responses, along with increased local IFN-γ and IgA levels compared to both the s.c. prime alone and the homologous oral prime-boost immunization. However, due to the relatively weak observed effect of the MC delivery on the immune responses, it was hypothesized that the MCs are proportionally too large for the GI tract of mice, and thus cleared before an effective immune response can be induced. To investigate this, MCs were loaded with BaSO4, and orally administered to mice. Analysis with X-ray and CT showed a transit time of approximately 1-1.5 h from the stomach to the cecum, corresponding to the standard transit time in mice, and an extremely narrow absorption window. This indicates that mice is not a suitable animal model for evaluation of MCs. These data should be taken into consideration in future in vivo trials with this and similar technologies, where larger animals might be a necessity for proof-of-concept studies.
Assuntos
Galactosilceramidas , Imunidade nas Mucosas , Animais , Camundongos , Galactosilceramidas/farmacologia , Vacinação , Adjuvantes Imunológicos , Adjuvantes Farmacêuticos/farmacologia , Chlamydia trachomatis , Vacinas de Subunidades Antigênicas , Camundongos Endogâmicos BALB CRESUMO
During the past decades, microdevices have been evaluated as a means to overcome challenges within oral drug delivery, thus improving bioavailability. Fabrication of microdevices is often limited to planar or simple 3D designs. Therefore, this work explores how microscale stereolithography 3D printing can be used to fabricate radiopaque microcontainers with enhanced mucoadhesive geometries, which can enhance bioavailability by increasing gastrointestinal retention. Ex vivo force measurements suggest increased mucoadhesion of microcontainers with adhering features, such as pillars and arrows, compared to a neutral design. In vivo studies, utilizing planar X-ray imaging, show the time-dependent gastrointestinal location of microcontainers, whereas computed tomography scanning and cryogenic scanning electron microscopy reveal information about their spatial dynamics and mucosal interactions, respectively. For the first time, the effect of 3D microdevice modifications on gastrointestinal retention is traced in vivo, and the applied methods provide a much-needed approach for investigating the impact of device design on gastrointestinal retention.
Assuntos
Sistemas de Liberação de Medicamentos , Tomografia Computadorizada por Raios X , Sistemas de Liberação de Medicamentos/métodos , Disponibilidade Biológica , Microscopia Eletrônica de Varredura , Impressão TridimensionalRESUMO
For patients with ulcerative colitis (UC), administration of the probiotic E. coli Nissle (EcN) holds promise for alleviation of disease symptoms. The mechanisms are unclear, but it has been hypothesised that a capacity of the probiotic to outcompete potentially detrimental UC-associated E. coli strains plays an important role. However, this could previously not be confirmed in a mouse model of competition between EcN and two UC-associated strains, as reported by Petersen et al. 2011. In the present study, we re-evaluated the idea, hypothesising that delivery of EcN by a micro device dosing system (microcontainers), designed for delivery into the intestinal mucus, could support colonisation and confer a competition advantage compared to classical oral dosing. Six groups of mice were pre-colonised with one of two UC-associated E. coli strains followed by oral delivery of EcN, either in capsules containing microcontainers with freeze-dried EcN powder, capsules containing freeze-dried EcN powder, or as a fresh sucrose suspension. Co-colonisation between the probiotic and the disease-associated strains was observed regardless of dosing method, and no competition advantages linked to microcontainer delivery were identified within this setup. Other approaches are thus needed if the competitive capacity of EcN in the gut should be improved.
Assuntos
Colite Ulcerativa , Probióticos , Humanos , Camundongos , Animais , Colite Ulcerativa/induzido quimicamente , Escherichia coli , PósRESUMO
In the pharmaceutical field, oral drug delivery devices continue to shrink down to the micrometer scale, driving a trending demand to investigate ex vivo mucoadhesive force down to the micro-Newton scale. However, owing to the limitation of measuring sensitivity, conventional methods (e.g., a texture analyzer) lack reliability while measuring forces in this range. Herein, we report on an open-source force analyzer that utilizes an optical-pickup-unit (from a DVD player) to detect cantilever-based force transducers and thereby, achieves a wide force-sensing range from 1.1 N to 0.99 nN. The cantilever force transducers can easily be adjusted to fit different force ranges by adjusting the steel shim, magnets, and 3D printed components. To validate the analyzer, we conducted a preliminary study to investigate the effect of time and humidity of mucoadhesion of porcine intestinal tissues. Besides, we measured the mucoadhesive force of a single oral drug delivery microdevice with an average force of 93.7 µN on the top sides of the device. This analyzer offers the possibility of measuring e.g. mucoadhesion of individual microdevices in the micro-Newton range. Hence, the analyzer can assist in the development of miniaturized oral drug delivery devices but has a much wider field of potential force sensing applications.
RESUMO
The biggest challenge in oral delivery of anti-inflammatory drugs such as 5-aminosalicylic acid (5-ASA) is to (i) prevent rapid absorption in the small intestine and (ii) achieve localized release at the site of inflammation in the lower gut, i.e., the colon. Here, we present an advanced biopolymeric coating comprising of tannic-acid-functionalized zein protein to provide a sustained, colon-targeted release profile for 5-ASA and enhance the mucoadhesion of the dosage form via a mussel-inspired mechanism. To enable localized delivery and provide high local concentration, 5-ASA is loaded into the microfabricated drug carriers (microcontainers) and sealed with the developed coating. The functionality and drug release profile of the coating are characterized and optimized in vitro, showing great tunability, scalability, and stability toward proteases. Further, ex vivo experiments demonstrate that the tannic acid functionalization can significantly enhance the mucoadhesion of the coating, which is followed up by in vivo investigations on the intestinal retention, and pharmacokinetic evaluation of the 5-ASA delivery system. Results indicate that the developed coating can provide prolonged colonic delivery of 5-ASA. Therefore, the here-developed biodegradable coating can be an eco-friendly substitute to the state-of-the-art commercial counterparts for targeted delivery of 5-ASA and other small molecule drugs.
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Now-a-days healthcare systems face great challenges with antibiotic resistance and low efficacy of antibiotics when combating pathogenic bacteria and bacterial biofilms. Administration of an antibiotic in its free form is often ineffective due to lack of selectivity to the infectious site and breakdown of the antibiotic before it exerts its effect. Therefore, polymeric delivery systems, where the antibiotic is encapsulated into a formulation, have shown great promise, facilitating a high local drug concentration at the site of infection, a controlled drug release and less drug degradation. All this leads to improved therapeutic effects and fewer systemic side effects together with a lower risk of developing antibiotic resistance. Here, we review and provide a comprehensive overview of polymer-based nano- and microparticles as carriers for antimicrobial agents and their effect on eradicating bacterial biofilms. We have a main focus on polymeric particulates containing poly(lactic-co-glycolic acid), chitosan and polycaprolactone, but also strategies involving combinations of these polymers are included. Different production techniques are reviewed and important parameters for biofilm treatment are discussed such as drug loading capacity, control of drug release, influence of particle size and mobility in biofilms. Additionally, we reflect on other promising future strategies for combating biofilms such as lipid-polymer hybrid particles, enzymatic biofilm degradation, targeted/triggered antibiotic delivery and future alternatives to the conventional particles.
Assuntos
Antibacterianos/administração & dosagem , Biofilmes/efeitos dos fármacos , Sistemas de Liberação de Medicamentos , Animais , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Portadores de Fármacos/química , Farmacorresistência Bacteriana , Humanos , Microesferas , Nanopartículas , Tamanho da Partícula , Polímeros/químicaRESUMO
Two simple, mechanical modifications are introduced to a consumer-grade inkjet printer to greatly increase its applicability. First, roller isolation bars are added to unlock multiple prints on the same substrate without smearing. This enables printing on a diverse set of substrates (rigid, elastic, liquid, granular, and sticky). Second, spring loadings are added to increase the print precision up to 50-fold, which facilitates alignment to a pre-patterned substrate or between successive prints. Utilizing the expanded substrate compatibility and the increased print precision, we explore tunable loading of drug combinations into microdevices. This loading method has promising applications within point-of-care personalized medication. Furthermore, we show how inkjet printers with array-type printheads (in our case, 6 x 90 nozzles) allow for quasi-simultaneous loading of reactants into microfluidic systems. The ability to do a quasi-simultaneous introduction of chemicals may be particularly useful for studies of rapidly reacting systems of three or more reactants, where premature introduction can shift the initial conditions from the intended. We believe that our modifications to an affordable system will inspire researchers to explore the possibilities of inkjet printing even further.
RESUMO
Numerous beneficial microbes thrive in the oral cavity where they form biofilms on dental and mucosal surfaces to get access to nutrients, and to avoid being carried away with the saliva. However, biofilm formation is also a virulence factor as it also protects pathogenic bacteria, providing them with an environment for proliferation causing oral infections. Oral hygiene relies on mechanical removal of biofilms. Some oral care products also contain antimicrobials, but effective eradication of biofilms with antimicrobials requires both a high concentration and long exposure time. In the present communication, we investigate the potential of using miniaturized drug delivery devices, known as microcontainers (MCs), to deliver the antimicrobial peptide, nisin to an oral multi-species biofilm. MCs are loaded with nisin and X-ray micro-computed tomography reveals a full release of nisin through a chitosan lid within 15 min. Chitosan-coated MCs display substantial bioadhesion to the buccal mucosa compared to non-coated MCs (68.6 ± 14.3% vs 33.8 ± 5.2%). Confocal monitoring of multi-species biofilms reveals antibacterial effects of nisin-loaded chitosan-coated MCs with a faster onset (after 3 h) compared to solution-based delivery (after 9 h). Our study shows the potential of using MCs for treatment of multi-species oral biofilms and is encouraging for further design of drug delivery devices to treat oral diseases.
Assuntos
Antibacterianos/administração & dosagem , Biofilmes/efeitos dos fármacos , Sistemas de Liberação de Medicamentos , Nisina/administração & dosagem , Adesivos , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Quitosana/química , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Humanos , Mucosa Bucal/metabolismo , Nisina/química , Nisina/farmacologia , Tamanho da Partícula , Suínos , Microtomografia por Raio-XRESUMO
Bacterial biofilm-related infections are difficult to eradicate and require repeated treatments with high doses of antibiotics. Thus, there is an urgent need for new treatment strategies that minimize the use of antibiotics while enhancing biofilm eradication. Functionalized reservoir-based microdevices, such as, microcontainers (MCs), offer, high drug loading capacity, mucus embedment, and tuneable drug release. Here, MCs are loaded with the antibiotic ciprofloxacin (CIP), and sealed with a lid consisting of chitosan (CHI) and a mucolytic agent, N-acetylcysteine (NAC). It is found that CHI and NAC work synergistically, showing improved mucoadhesive and mucolytic properties. To better mimic the in vivo habitat of Pseudomonas aeruginosa (P. aeruginosa), the biofilm is grown in a mucin-containing medium on a newly developed centrifugal microfluidic system. The CHI/NAC coated MCs improve eradication of biofilm (88.22 ± 2.89%) compared to CHI-coated MCs (72.68 ± 3.73%) or bolus injection (39.86 ± 13.28%). The findings suggest that MCs are significantly more efficient than a bolus treatment. Furthermore, CHI/NAC functionalized MCs kill most of the biomass already after 5 h (80.75 ± 3.50%), mainly due to a fast drug release. This is the first time that CHI/NAC has been combined as a coating to explore mucolytic properties on bacterial biofilms.
Assuntos
Antibacterianos , Mucinas , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Biofilmes , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosaRESUMO
Microcontainers, which are microfabricated cylindrical devices with a reservoir function, have shown promise as an oral drug delivery system for small molecular drug compounds. However, they have never been evaluated against a relevant control formulation. In the current study, we prepared microcrystalline cellulose (MCC) microspheres as a control for in vitro and in vivo testing of SU-8 microcontainers as an oral drug delivery system. Both dosage forms were loaded with paracetamol and coated with chitosan or polyethylene glycol (PEG) (12 kDa). These coatings were followed by an additional enteric coating of Eudragit® S100. In addition, a control dosage form was coated with Eudragit® alone. The dosage forms were evaluated in vitro, in a physiologically relevant two-step model simulating rat gastrointestinal fluids, and in vivo by oral administration to rats. In vitro, the microcontainers coated with PEG/Eudragit® resulted in a prolonged release of paracetamol compared to the respective microspheres, which was consistent with in vivo observations of a later time (Tmax) for maximum plasma concentration (Cmax) for the microcontainers. For microspheres and microcontainers coated with chitosan/Eudragit®, the time for complete in vitro release of paracetamol was very similar, due to an earlier release from the microcontainers. This trend was supported by very similar Tmax values in vivo. The in vitro in vivo relation was confirmed by a linear regression with R2 = 0.9, when Tmax for each dosage form was plotted as a function of time for 90% paracetamol release in vitro. From the in vivo study, the average plasma concentration of paracetamol 120 min after dosing was significantly higher for microcontainers than for microspheres (0.3 ± 0.1 µg/mL and 0.1 ± <0.1 µg/mL, respectively) - regardless of the coating applied.
Assuntos
Quitosana , Preparações Farmacêuticas , Administração Oral , Animais , Sistemas de Liberação de Medicamentos , Microesferas , Ácidos Polimetacrílicos , RatosRESUMO
Microscale devices are promising tools to overcome specific challenges within oral drug delivery. Despite the availability of advanced high-quality imaging techniques, visualization and tracking of microscale devices in the gastrointestinal (GI) tract is still a challenge. This work explores the possibilities of applying planar X-ray imaging and computed tomography (CT) scanning for visualization and tracking of microscale devices in the GI tract of rats. Microcontainers (MCs) are an example of microscale devices that have shown great potential as an oral drug delivery system. Barium sulfate (BaSO4) loaded into the cavity of the MCs increases their overall X-ray contrast, which allows them to be easily tracked. The BaSO4-loaded MCs are quantitatively tracked throughout the entire GI tract of rats by planar X-ray imaging and visualized in 3D by CT scanning. The majority of the BaSO4-loaded MCs are observed to retain in the stomach for 0.5-2 h, enter the cecum after 3-4 h, and leave the cecum and colon 8-10 h post-administration. The imaging approaches can be adopted and used with other types of microscale devices when investigating GI behavior in, for example, preclinical trials and potential clinical studies.