Natural biopolymer nanoparticles (NPs), including nanocrystalline cellulose (CNC) and lignin, have shown potential as scaffolds for targeted drug delivery systems due to their wide availability, cost‐efficient preparation, and anticipated biocompatibility. As both CNC and lignin can potentially cause complications in cell viability assays because of their ability to scatter the emitted light and absorb the assay reagents, we investigated the response of bioluminescent (CellTiter‐Glo®), colorimetric (MTT® and AlamarBlue®), and fluorometric (LIVE/DEAD®) assays for the determination of the biocompatibility of the multimodal CNC and lignin constructs in murine RAW 264.7 macrophages and 4T1 breast adenocarcinoma cell lines. Here, we have developed multimodal CNC and lignin NPs harboring the radiometal chelator 1,4,7,10‐tetraazacyclododecane‐1,4,7,10‐tetraacetic acid and the fluorescent dye cyanine 5 for the investigation of nanomaterial biodistribution in vivo with nuclear and optical imaging, which were then used as the model CNC and lignin nanosystems in the cell viability assay comparison. CellTiter‐Glo® based on the detection of ATP‐dependent luminescence in viable cells revealed to be the best assay for both nanoconstructs for its robust linear response to increasing NP concentration and lack of interference from either of the NP types. Both multimodal CNC and lignin NPs displayed low cytotoxicity and favorable interactions with the cell lines, suggesting that they are good candidates for nanosystem development for targeted drug delivery in breast cancer and for theranostic applications. Our results provide useful guidance for cell viability assay compatibility for CNC and lignin NPs and facilitate the future translation of the materials for in vivo applications.

中文翻译：

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多峰纤维素纳米晶体和木质素纳米颗粒的系统体外生物相容性研究。

天然生物聚合物纳米颗粒 (NPs)，包括纳米晶纤维素 (CNC) 和木质素，由于其广泛的可用性、成本效益的制备和预期的生物相容性，已显示出作为靶向给药系统支架的潜力。由于 CNC 和木质素具有散射发射光和吸收测定试剂的能力，因此它们都可能导致细胞活力测定的并发症，因此我们研究了生物发光 (CellTiter-Glo®)、比色法 (MTT® 和 AlamarBlue®) 的响应，和荧光 (LIVE/DEAD®) 测定法，用于确定小鼠 RAW 264.7 巨噬细胞和 4T1 乳腺癌细胞系中多模式 CNC 和木质素构建体的生物相容性。在这里，我们开发了含有放射性金属螯合剂 1,4,7,10-四氮杂环十二烷-1,4,7 的多峰 CNC 和木质素 NP，10-四乙酸和荧光染料花青 5 用于通过核和光学成像研究体内纳米材料的生物分布，然后在细胞活力测定比较中用作模型 CNC 和木质素纳米系统。CellTiter-Glo® 基于对活细胞中 ATP 依赖性发光的检测表明是两种纳米结构的最佳检测方法，因为它对增加的 NP 浓度具有稳健的线性响应，并且不受任何一种 NP 类型的干扰。多模式 CNC 和木质素 NPs 均显示出低细胞毒性和与细胞系良好的相互作用，表明它们是用于乳腺癌靶向药物递送和治疗诊断应用的纳米系统开发的良好候选者。