Assessing biologic/toxicologic effects of extractables from plastic contact materials for advanced therapy manufacturing using cell painting assay and cytotoxicity screening

Abstract

Plastic components are essential in the pharmaceutical industry, encompassing container closure systems, laboratory handling equipment, and single-use systems. As part of their material qualification process, studies on interactions between plastic contact materials and process solutions or drug products are conducted. The assessment of single-use systems includes their potential impact on patient safety, product quality, and process performance. This is particularly crucial in cell and gene therapy applications since interactions with the plastic contact material may result in an adverse effect on the isolated therapeutic human cells. We utilized the cell painting assay (CPA), a non-targeted method, for profiling the morphological characteristics of U2OS human osteosarcoma cells in contact with chemicals related to plastic contact materials. Specifically, we conducted a comprehensive analysis of 45 common plastic extractables, and two extracts from single-use systems. Results of the CPA are compared with a standard cytotoxicity assay, an osteogenesis differentiation assay, and in silico toxicity predictions. The findings of this feasibility study demonstrate that the device extracts and most of the tested compounds do not evoke any measurable biological changes on the cells (induction ≤ 5%) among the 579 cell features measured at concentrations ≤ 50 µM. CPA can serve as an important assay to reveal unique information not accessible through quantitative structure–activity relationship analysis and vice versa. The results highlight the need for a combination of in vitro and in silico methods in a comprehensive assessment of single-use equipment utilized in advanced therapy medicinal products manufacturing.

Parylene C Coating Efficacy Studies: Enhancing Biocompatibility of 3D Printed Polyurethane Parts for Biopharmaceutical and CGT Applications

“…47 It should be noted that TPO was recently flagged as a compound to show an induction to U2OS human cells detected by a cell painting assay. 48 The low EC 50 value for CHO cells, together with reported cytotoxicity of the compound in literature, supports the hypothesis that TPO, and not possible other extractables compounds (e.g., monomers), is the primary contributor to reduced cell growth in cell cultures in contact with components made from photopolymers that incorporate TPO as a photoinitiator. 3.7.…”

“…This value is significantly lower, almost an order of magnitude, compared to those reported for other cell lines, such as human oral keratinocytes and V79 cells, or MRC-5 human fetal lung fibroblasts, when tested with different assay systems. , The higher sensitivity observed in the ASTM E3231 test is likely due to the use of serum-free cell culture media, a phenomenon that has been documented in other tests as well . It should be noted that TPO was recently flagged as a compound to show an induction to U2OS human cells detected by a cell painting assay . The low EC 50 value for CHO cells, together with reported cytotoxicity of the compound in literature, supports the hypothesis that TPO, and not possible other extractables compounds (e.g., monomers), is the primary contributor to reduced cell growth in cell cultures in contact with components made from photopolymers that incorporate TPO as a photoinitiator.…”