RESUMO
Medical device biocompatibility testing is used to evaluate the risk of adverse effects on tissues from exposure to leachates/extracts. A battery of tests is typically recommended in accordance with regulatory standards to determine if the device is biocompatible. In vitro cytotoxicity, a key element of the standards, is a required endpoint for all types of medical devices. Each validated cytotoxicity method has different methodology and acceptance criteria that could influence the selection of a specific test. In addition, some guidances are more specific than others as to the recommended test methods. For example, the International Organization for Standardization (ISO1) cites preference for quantitative methods (e.g., tetrazolium (MTT/XTT), neutral red (NR), or colony formation assays (CFA)) over qualitative methods (e.g., elution, agar overlay/diffusion, or direct), while a recent ISO standard for contact lens/lens care solutions specifically requires a qualitative direct test. Qualitative methods are described in United States Pharmacopeia (USP) while quantitative CFAs are listed in Japan guidance. The aim of this review is to compare the methodologies such as test article preparation, test conditions, and criteria for six cytotoxicity methods recommended in regulatory standards in order to inform decisions on which method(s) to select during the medical device safety evaluation.
Assuntos
Materiais Biocompatíveis/farmacologia , Segurança de Equipamentos , Teste de Materiais , Animais , Sobrevivência Celular/efeitos dos fármacos , HumanosRESUMO
Purpose: A new class of ocular steroids designed to mitigate steroid-induced intraocular pressure (IOP) elevation while maintaining anti-inflammatory activity was developed. Herein is described the discovery and preclinical characterization of ROCK'Ster compound 1. Methods: Codrugs consisting of a Rho kinase inhibitor (ROCKi) and a corticosteroid were synthesized. Compounds were initially screened in vitro for ROCKi activity and anti-inflammatory activity against the proinflammatory interleukin 23 and bacterial lipopolysaccharide (LPS) pathways. Selected compounds were then screened for solubility, chemical stability, and ex vivo corneal metabolism. Lead compound 1 was evaluated for IOP lowering in the Dutch Belted rabbit and for anti-inflammatory efficacy in both a postcataract surgery model and an allergic eye disease (AED) mouse model. Results: Several ROCK'Sters were found to be potent inhibitors of ROCK (Kis < 50 nM), have high anti-inflammatory activity in vitro (IC50s < 50 nM), display sufficient stability in topical ophthalmic formulations, and have a moderate rate of corneal metabolism. Compound 1 (0.1% and 0.25%, quater in die [QID]-4 times a day) demonstrated IOP-lowering capability without inducing hyperemia in our rabbit model. When compared with the marketed steroids, Durezol® and Pred Forte®, compound 1 (0.1%, 0.25%) demonstrated noninferiority in clinical scoring in a rabbit model of inflammation after surgery. In addition, anti-inflammatory outcomes were observed with compound 1 (0.1%) relative to Lotemax® or vehicle control in an AED mouse model. Conclusion: ROCK'Ster compound 1 is a novel compound suitable for topical ocular dosing that possesses IOP-lowering capability along with similar anti-inflammatory activity compared with marketed steroids.
Assuntos
Oftalmopatias , Pressão Intraocular , Animais , Camundongos , Coelhos , Quinases Associadas a rho , Oftalmopatias/tratamento farmacológico , Modelos Animais de Doenças , Corticosteroides , Córnea , Soluções Oftálmicas/farmacologia , Anti-HipertensivosRESUMO
An unmet medical need remains for patients suffering from dry eye disease (DED). A fast-acting, better-tolerated noncorticosteroid anti-inflammatory eye drop could improve patient outcomes and quality of life. Herein, we describe a small-molecule drug discovery effort to identify novel, potent, and water-soluble JAK inhibitors as immunomodulating agents for topical ocular disposition. A focused library of known 3-(4-(2-(arylamino)pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitriles was evaluated as a molecular starting point. Structure-activity relationships (SARs) revealed a ligand-efficient (LE) JAK inhibitor series, amenable to aqueous solubility. Subsequent in vitro analysis indicated the potential for off-target toxicity. A KINOMEscan selectivity profile of 5 substantiated the likelihood of widespread series affinity across the human kinome. An sp2-to-sp3 drug design strategy was undertaken to attenuate off-target kinase activity while driving JAK-STAT potency and aqueous solubility. Tactics to reduce aromatic character, increase fraction sp3 (Fsp3), and bolster molecular complexity led to the azetidin-3-amino bridging scaffold in 31.
Assuntos
Inibidores de Janus Quinases , Humanos , Janus Quinase 1 , Janus Quinase 2 , Janus Quinase 3 , Inibidores de Janus Quinases/farmacologia , Janus Quinases , Fosforilação , Inibidores de Proteínas Quinases/farmacologia , SolubilidadeRESUMO
We used published accounts to describe the known statewide distribution of the lone star tick, Amblyomma americanum, in New Jersey and field surveys to characterize the geographical range of A. americanum and selected A. americanum-transmitted pathogens in Monmouth County, the hypothesized northern limit of the species distribution. Ticks were collected using standardized methods from 50 widely dispersed public access areas within 18 municipalities to produce estimates of relative abundance among sites. Collected A. americanum adults were stored at -80 degrees C until processed for DNA extraction. Individual ticks were subjected to polymerase chain reaction analyses to detect the presence of Ehrlichia chaffeensis, E. ewingii, Rickettsia amblyommii, and Borrelia lonestari. The range of A. americanum was generally limited to the southern half of New Jersey. Within Monmouth County, we collected A. americanum from 9 of 18 municipalities (50%) and 24 (48%) of the surveyed properties. We found at least 1 pathogen at 17 (70.8%) sites located within 6 of 9 municipalities, while all 4 target pathogens were detected in 5 of those 6 (83.3%) municipalities. The geographical distribution of A. americanum and its associated pathogens appeared to be restricted to the southern portion of the county. Possible factors governing the distribution are discussed.
Assuntos
Borrelia/isolamento & purificação , Ehrlichia/isolamento & purificação , Ixodidae/microbiologia , Rickettsia/isolamento & purificação , Animais , Infecções por Borrelia/epidemiologia , Infecções por Borrelia/transmissão , Ehrlichiose/epidemiologia , Ehrlichiose/transmissão , Geografia , New Jersey/epidemiologia , Densidade Demográfica , Prevalência , Infecções por Rickettsia/epidemiologia , Infecções por Rickettsia/transmissãoRESUMO
Vitamin A plays an essential role in vertebrate embryogenesis. In the present study, pregnant vitamin A-deficient (VAD) rats were maintained during early pregnancy on the short half-life vitamin A metabolite, all-trans retinoic acid (atRA), in an amount sufficient to support normal development to E10.5, with a higher level of atRA (250 microg atRA/g diet) provided from embryonic day (E) 8.5-10.5 to prevent mid-gestational resorption. When limiting amounts of atRA (1.5 or 12 microg/g diet) were provided after E10.5, a highly reproducible and penetrant state of late fetal vitamin A deficiency (late VAD) was induced in the organs of developing fetuses. In addition, late VAD fetuses displayed both anteriorization of cervical regions and novel posteriorization events at the thoracic and sacral levels of the skeleton, and showed sternal and pelvic malformations not previously observed in early VAD or genetic models. The expression of several Hox genes (Hoxd3 and Hoxb4) was altered in late VAD embryos, with a reduction in Hoxd3 noted as early as 1 day after instituting deficiency. All late VAD-induced malformations were prevented by the addition of retinol starting at E10.5, whereas provision of a high level of atRA throughout pregnancy improved but could not completely rescue the development of all organ systems. This work defines a nutritional model in which vitamin A deficiency can be induced during fetal development, and reveals new functions for the vitamin in the development of the axial and appendicular skeleton.
Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Desenvolvimento Ósseo/fisiologia , Desenvolvimento Fetal/fisiologia , Organogênese/fisiologia , Deficiência de Vitamina A/embriologia , Animais , Modelos Animais de Doenças , Gástrula/fisiologia , Hibridização In Situ , Ratos , Ratos Sprague-Dawley , Tretinoína/uso terapêutico , Deficiência de Vitamina A/tratamento farmacológicoRESUMO
The effects of soil moisture and temperature on the reproduction of twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), were examined in laboratory and field tests in strawberries, Fragaria x ananassa Duchesne, in Florida. Different soil moisture levels (low, moderate, and high) were compared to determine how soil moisture affects the reproduction and development of twospotted spider mite. In addition to soil moisture, different irrigation techniques (drip versus drip/overhead) were compared to determine their effects on twospotted spider mite reproduction as well as the incidence of angular leaf spot, Xanthomonas fragaria Kennedy & King disease. Similar studies were conducted to determine how different temperatures (18, 27, and 35 degrees C) affect the reproduction and development of twospotted spider mites. In the laboratory, low soil moisture as well as temperatures >27 degrees C promoted twospotted spider mite development. A similar trend was observed in a field study with low soil moisture promoting twospotted spider mite reproduction during the early season (11 November--8 December). Irrespective of moisture levels, a significantly higher incidence of X. fragaria was recorded in treatments with drip/overhead irrigation systems compared with drip irrigation. Implications for management of soil moisture levels are discussed with respect to the abundance of twospotted spider mite and X. fragaria in strawberries.
Assuntos
Fragaria , Solo/análise , Temperatura , Tetranychidae/crescimento & desenvolvimento , Água/análise , Agricultura/métodos , Animais , Fragaria/microbiologia , Controle de Insetos/métodos , Doenças das Plantas/microbiologia , Reprodução , XanthomonasRESUMO
Disposable diapers are primarily composed of polymers, such as cellulose, polypropylene, polyester, and polyethylene, which are biologically inert and not bioavailable. They are used in clothes, fabrics, personal hygiene products, and other materials that are commonly in contact with the skin. Each component used throughout the production process must undergo rigorous safety evaluations and assessments and are proven to be well tolerated and safe for their intended uses. No materials are incorporated into a diaper until their safety is confirmed through robust assessments, and additional factors are integrated into the process to compensate for the uncertainty associated with extrapolating toxicity data. After a thorough assessment of the materials and final product, extensive skin compatibility evaluations are conducted as appropriate. This rigorous safety process provides reassurance that consumers can rely on the safety of these diapers.
RESUMO
Skin in the diapered area is continuously threatened by exposure to changes in pH levels, overhydration, mechanical friction, and fecal enzymes, making diaper rash a common occurrence among babies. Up to one third of infants may exhibit clinical symptoms of diaper rash at any time, and more than half of babies between the ages of 4 and 15 months develop diaper rash at least once in a 2-month period. Despite misperceptions that disposable diapers are related to an increase in diaper rash, the incidence of diaper dermatitis is on the decline, largely due to significant improvements in disposable diaper construction and materials. Modern-day disposable diapers are specifically designed to limit exposure to irritants in the diaper area, reduce overhydration, inhibit skin barrier compromise, and help maintain normal skin pH levels and have been thoroughly evaluated for safety and skin compatibility.