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开发用于使用 cAMP 作为模型物质表征肿瘤内注射剂的 UV 成像的方法学考虑因素。

Methodological Considerations in Development of UV Imaging for Characterization of Intra-Tumoral Injectables Using cAMP as a Model Substance.

机构信息

Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.

Bristol Myers Squibb Company, Drug Product Development, 1 Squibb Drive, New Brunswick, NJ 08901, USA.

出版信息

Int J Mol Sci. 2022 Mar 25;23(7):3599. doi: 10.3390/ijms23073599.

DOI:10.3390/ijms23073599
PMID:35408971
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8998202/
Abstract

A UV imaging release-testing setup comprising an agarose gel as a model for tumorous tissue was developed. The setup was optimized with respect to agarose concentration (0.5% (/)), injection procedure, and temperature control. A repeatable injection protocol was established allowing injection into cavities with well-defined geometries. The effective resolution of the SDi2 UV imaging system is 30-80 µm. The linear range of the imaging system is less than that of typical spectrophotometers. Consequently, non-linear cAMP calibration curves were applied for quantification at 280 nm. The degree of deviation from Beer's law was affected by the background absorbance of the gel matrix. MATLAB scripts provided hitherto missing flexibility with respect to definition and utilization of quantification zones, contour lines facilitating visualization, and automated, continuous data analysis. Various release patterns were observed for an aqueous solution and in situ forming Pluronic F127 hydrogel and PLGA implants containing cAMP as a model for STING ligands. The UV imaging and MATLAB data analysis setup constituted a significant technical development in terms of visualizing behavior for injectable formulations intended for intra-tumoral delivery, and, thereby, a step toward establishment of a bio-predictive in vitro release-testing method.

摘要

开发了一种包含琼脂糖凝胶作为肿瘤组织模型的 UV 成像释放测试装置。该装置在琼脂糖浓度（0.5% (/))、注射程序和温度控制方面进行了优化。建立了可重复的注射方案，允许在具有明确定义几何形状的腔室内进行注射。SDi2 UV 成像系统的有效分辨率为 30-80 µm。成像系统的线性范围小于典型分光光度计的线性范围。因此，在 280nm 处应用了非线性 cAMP 校准曲线进行定量。偏离比尔定律的程度受凝胶基质的背景吸收的影响。MATLAB 脚本提供了迄今为止在定义和利用定量区域、便于可视化的等高线以及自动、连续数据分析方面缺失的灵活性。观察到水溶液以及原位形成的 Pluronic F127 水凝胶和含有 cAMP 的 PLGA 植入物的各种释放模式，cAMP 作为 STING 配体的模型。UV 成像和 MATLAB 数据分析装置在可视化用于肿瘤内递送的可注射制剂的行为方面取得了重大技术进展，从而朝着建立生物预测性体外释放测试方法迈出了一步。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f2/8998202/c137fa886367/ijms-23-03599-g009.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f2/8998202/c137fa886367/ijms-23-03599-g009.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f2/8998202/e01a8ffa5f80/ijms-23-03599-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f2/8998202/17b9de645048/ijms-23-03599-g007.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04f2/8998202/c137fa886367/ijms-23-03599-g009.jpg

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