119th General Meeting of the KCS

Type Oral Presentation
Area Oral Presentation of Young Analytical Chemists Ⅱ
Room No. 303호
Time FRI 10:00-:
Code ANAL2.O-27
Subject Flow cytometry study of the influence of agglomeration on effective in vitro dosimetry and cellular association of silver nanoparticles
Authors HA KIEU MY, 심유진, 윤태현*
한양대학교 화학과, Korea
Abstract Nanoparticles (NPs) are becoming increasingly important in various biological and medical applications, from imaging and diagnostics to therapeutics and regenerative medicine, which requires comprehensive knowledge on how NPs interact with biological systems. In vitro assays are simple, fast and inexpensive approach to investigate these interactions and have been very useful in providing insights into the cellular interaction of NPs. However, they still face several challenges regarding their accuracy and reproducibility. One of the challenges is the effective dosimetry for inhomogeneously dispersed chemicals, such as NP dispersions. The actual NP concentrations that reach the adherent cells’ surface are often different from the administered value, and one of the reasons for this discrepancy is the agglomeration and sedimentation of NPs. Exposure of NPs to cell culture media typically induce agglomeration and cause changes in their effective size and density. These parameters play important roles in their sedimentation and diffusion processes, which strongly influence their effective dose and cellular associations. Therefore, it is necessary to study how agglomeration of NPs affects their effective dose and cellular association, so that accurate and reproducible assay results can be generated. The objective of this study is to investigate the influence of Ag NP agglomeration on their effective in vitro dosimetry and their association with human lung cancer cells (A549). To address the influences of NP agglomeration on effective in vitro dosimetry, hydrodynamic size and effective density of Ag NP agglomerates in cell culture media were measured by dynamic light scattering (DLS) and volumetric centrifugation method (VCM) respectively, and were then used to estimate the effective in vitro dose of NP dispersions using a computational model developed by DeLoid et al. Moreover, to evaluate the impact of NP agglomeration on the sedimentation and diffusion processes, A549 cells were exposed to Ag NPs in upright and inverted configurations. Flow cytometry (FCM) side scattering (SSC) intensity measurement was used to quantify cellular association of Ag NPs. In the FCM technique, the SSC signal, which is related to the inner complexity of the cells, was combined with inductively coupled plasma mass spectrometry (ICPMS) technique and used to evaluate the association level of Ag NPs in A549 cells. On the basis of these FCM SSC and ICPMS measurements, we demonstrated the influence of agglomeration on the sedimentation and diffusion processes of NPs, and thus on effective in vitro dosimetry and cellular association of Ag NPs.
E-mail hakieumy12@gmail.com