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    Low-cost, rapidly-developed, 3D printed in vitro corpus callosum model for mucopolysaccharidosis type I
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    Abstract:
    The rising prevalence of high throughput screening and the general inability of (1) two dimensional (2D) cell culture and (2)in vitrorelease studies to predictin vivoneurobiological and pharmacokinetic responses in humans has led to greater interest in more realistic three dimensional (3D) benchtop platforms. Advantages of 3D human cell culture over its 2D analogue, or even animal models, include taking the effects of microgeometry and long-range topological features into consideration. In the era of personalized medicine, it has become increasingly valuable to screen candidate molecules and synergistic therapeutics at a patient-specific level, in particular for diseases that manifest in highly variable ways. The lack of established standards and the relatively arbitrary choice of probing conditions has limitedin vitrodrug release to a largely qualitative assessment as opposed to a predictive, quantitative measure of pharmacokinetics and pharmacodynamics in tissue. Here we report the methods used in the rapid, low-cost development of a 3D model of a mucopolysaccharidosis type I patient’s corpus callosum, which may be used for cell culture and drug release. The CAD model is developed fromin vivobrain MRI tracing of the corpus callosum using open-source software, printed with poly (lactic-acid) on a Makerbot Replicator 5X, UV-sterilized, and coated with poly (lysine) for cellular adhesion. Adaptations of material and 3D printer for expanded applications are also discussed.
    Цель исследования провести экспериментальную оценку гемостатической активности локальных покрытий в форме губки на основе растворов хитозана in vivo и in vitro. Материал и методы. Для изготовления исследуемых покрытий использовались растворы хитозана с массовой долей 0,5, 1,0, 1,5 и 2,0, в качестве растворителя применяли 0,5 уксусную или 0,5 молочную кислоту. На основе данных растворов были изготовлены экспериментальные образцы покрытий в форме губки. Гемостатическую активность определяли посредством измерения времени остановки кровотечения и объема кровопотери. Влияние взаимодействия контактной поверхности покрытий в форме губки с кровью in vitro оценивали по результатам подсчета числа тромбоцитов и определения уровня фибриногена в плазме крови. Интегральную оценку системы гемостаза проводили с помощью тромбоэластометрии стабилизированной цитратом натрия крови и теста генерации тромбина в плазме крови. Результаты. Высокой гемостатической активностью в экспериментах in vivo обладали локальные покрытия в форме губки на основе 0,5 и 2,0 растворов хитозана в 0,5 уксусной кислоте (67,6524,23 и 65,659,68 соответственно). Результаты исследований in vitro данных покрытий подтвердили ускорение процесса образования первичного тромба при взаимодействии их с кровью. Покрытия на основе изученных растворов хитозана в 0,5 молочной кислоте и 1,0 и 1,5 растворов хитозана в 0,5 уксусной кислоте не проявили гемостатической активности в экспериментах in vivo и практически не изменяли параметры гемостаза in vitro. Экспериментальные исследования 59 Клиническая физиология кровообращения. 2020 17 (1). DOI: 10.24022/1814-6910-2020-17-1-58-69 Заключение. Выявлена взаимосвязь между значениями гемостатической активности раневых покрытий на основе растворов хитозана в уксусной кислоте in vivo при их локальной аппликации на раневую поверхность печени кролика и особенностями реагирования крови при контакте с этими покрытиями in vitro. Проведение совместных исследований по изучению системы гемостаза как in vivo, так и in vitro представляется перспективным направлением для оценки взаимосвязи структурного состояния и механизма действия новых гемостатических средств.
    A qualitative comparison of the in vitro and in vivo activity on β-hemo1ytic streptococcus of 126 compounds consisting of sulfanilamide derivatives, sulfonamides, suifones, sulfoxides, sulfides and certain miscellaneous compounds has been made. It has been shown that no compound is active in vivo unless it is active in vitro or can be decomposed in the animal body to a compound which would be active in vitro , and that compounds can be active in vitro but inactive in vivo.
    Sulfanilamide
    Sulfonamide
    Active compound
    The endpoints of in vivo and in vitro assays applied to cells after exposure to a potential oncogenic transforming agent are cellular tumorigenicity and transformation. Tumors are failures of in vivo growth control; transformations are failures of in vitro growth control. Many agents cause tumors in vivo, many agents transform normal cultured cells, and some agents do both. However, even when caused by a single agent, the in vivo and in vitro endpoint assays show only a partial overlap. That is, some but not all tumors will grow as transformed cells in culture, and some but not all in vitro transformants will be tumorigenic on injection into susceptible animals (Shin et al., 1975). Recently, we have described a subset of in vitro phenotypic changes that correlate with in vivo tumorigenicity (Steinberg et al., 1979; Barrett et al., 1979; Pollack, 1981). In this chapter, we will describe recent studies on one of the in vitro changes linked to tumorigenicity, the disruption in organization of cyto-skeletal actin.
    To determine the optimal condition of the clonogenic assay, the antitumor activities of 5-fluorouracil (5-FU) in vitro and in vivo were investigated from a pharmacodynamic viewpoint using 8 human carcinoma xenografts. The clonogenic assay was performed by the continuous exposure method, and the antitumor effects were evaluated by the colony survival rates (T/Cs). The in vivo experimental chemotherapy was also performed by the nude mouse system, and the results were evaluated by the T/C ratios of the tumor weights. For pharmacokinetic analyses, the area under the concentration curves (AUCs) of 5-FU in vitro and in vivo were computed. The T/Cs of 5-FU were highly correlated to the AUCs both in vitro and in vivo. By using these AUC-T/C correlations, the concentration of 5-FU in the clonogenic assay to predict the T/C of the maximum tolerated dose in mouse was calculated to be 3 micrograms/ml mathematically. This concentration was then verified by the clonogenic assay, where T/Cs in vitro could successfully correspond to the T/Cs in vivo. (The predictable rate was 87.5%) From these results, this pharmacodynamic comparison between in vitro and in vivo chemosensitivities was thought to be a promising method for determining the conditions of the clonogenic assay.
    Clonogenic assay
    Pharmacodynamics
    In vitro toxicology
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    Selective tumor targeting strategies based on cell surface molecules enable new personalized diagnosis and treatments, potentially lowering adverse effects and increasing efficacy. Radio-immunotargeting generally relies on a molecule binding to a cancer-specific target. It is therefore important to understand the properties of molecular interactions in their working environment and how to translate these properties measured in vitro into the in vivo molecular imaging situation.Time resolved interaction analysis in vitro was compared with a corresponding in vivo xenograft mouse model. The antibody fragment AbD15179 was labeled with (125)I or (111)In, and analyzed on cell lines with differing CD44v6 expression in vitro, and in a dual tumor xenograft model derived from the same cell lines. In vitro LigandTracer measurements were analyzed with TraceDrawer and Interaction Map. Conjugate sensitivity, kinetics, and signal-to-background ratios were assessed for both tumor cells in vitro and xenograft tumors in vivo.In vitro results revealed a general biphasic appearance of a high- and a low-affinity interaction event. The (111)In-labeled fragment displayed the largest proportion of the high-affinity interaction with increased sensitivity and retention compared to (125)I-Fab. In vivo results were in agreement with in vitro data, with increased retention, higher sensitivity and better contrast for the (111)In-labeled fragment compared to (125)I.Time resolved binding characteristics measured in vitro largely matched the in vivo performance for the conjugates, which is promising for future studies. In vitro time-resolved LigandTracer assays are efficient, rapid, and in this study shown to be able to predict in vivo outcomes.Further studies are needed to confirm these findings, but the method is promising considering the ethical need to reduce the use of laboratory animals, as well as reducing costs for the development of tumor targeting compounds in the future.
    Conjugate
    The ointments with marked insulin J125 were prepared on suitable ointment bases. The dynamics of the released insulin from ointment in vitro and in vivo was analyzed. It has been noticed that the most of the marked insulin J125 was released in vitro and in vivo from eucerin and that there is a correlation between the amount of released insulin from ointment basis in vitro and in vivo.
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    SUMMARY Highly specific antibodies to mouse macrophages can be obtained in rabbits following immunization with pure cultures of mouse macrophages. Antimacrophage serum prepared in the manner described reacted specifically with macrophages in vitro and interfered with the initiation of the immune response to trinitrophenol sullonate in vivo.
    An in vitro method and an in vivo method of excystation were compared to determine the most useful method for the retrieval of Giardia duodenalis isolates. Cysts from 11 Giardia strains were used. In vitro excystation produced motile trophozoites in 16 sets, while in vivo excystation produced trophozoites in all of the 21 comparative sets of excystations. Few cultures were lost because of contamination by either method (17% of in vitro-derived trophozoites versus 23% of in vivo-derived trophozoites; P greater than 0.05). Both methods demonstrated comparable isolate retrieval rates (15% of in vitro-derived trophozoites adapting to culture compared with 29% of in vivo-derived trophozoites; P greater than 0.05), although analysis of the strains retrieved showed that two isolates were retrieved from in vitro excystation alone, compared with four from in vivo excystation. Analysis that included results of extra in vivo cultures showed that a total of nine isolates were retrieved by using this type of excystation. Despite the disadvantages of cost and labor, in vivo excystation appears to be more useful than in vitro excystation for isolate retrieval at the present time.
    Giardia
    Giardia lamblia