RESUMO
Three-dimensional (3D) cell culture based on polymer scaffold provides a promising tool to mimic a physiological microenvironment for drug testing; however, the next-generation cell activity monitoring technology for 3D cell culture is still challenging. Conventionally, drug efficacy evaluation and cell growth heavily rely on cell staining assays, using optical devices or flow cytometry. Here, we report a dual-function polymer scaffold (DFPS) composed of thermosensitive, silver flake- and gold nanoparticle-decorated polymers, enabling conductance change upon cell proliferation or death for in situ cell activity monitoring and drug screening. The cell activity can be quantitatively monitored via measuring the conductance change induced by polymeric network swelling or shrinkage. This novel dual-function system (1) provides a 3D microenvironment to enable the formation and growth of tumor spheroid in vitro and streamlines the harvesting of tumor spheroids through the thermosensitive scaffold and (2) offers a simple and direct quantitative method to monitor 3D cell culture in situ for drug responses. As a proof of concept, we demonstrated that a breast cancer stem cell line MDA-MB-436 was able to form cell spheroids in the scaffold, and the conductance change of the sensor exhibited a linear relationship with cell concentration. To examine its potential in drug screening, cancer spheroids in the cell sensor were treated with paclitaxel (PTX) and docetaxel (DTX), and predicted quantitative evaluation of the cytotoxic effect of drugs was established. Our results indicated that this cell sensing system may hold promising potential in expanding into an array device for high-throughput drug screening.
Assuntos
Nanopartículas Metálicas , Preparações Farmacêuticas , Ouro , Polímeros , Esferoides CelularesRESUMO
Ultra-small gold nanoclusters (AuNCs) have unique size-dependent optical, electrical and chemical properties. They have emerged as a new nanomaterial with broad applications in optoelectronics, catalysis, biosensing, and bioimaging. Several strategies have been exploited to prepare AuNCs of different "magic number" sizes, using different templates e.g. dendrimers, polyethyleneimines, peptides, and more recently, proteins. Notwithstanding, almost all bio-template-protected AuNCs reported so far exhibit fairly low fluorescence quantum yields (QYs), typically <5%, which is especially true for AuNCs prepared using the protein templates. In this paper, we report a facile, one-pot aqueous synthesis of highly fluorescent AuNCs using bovine pancreatic ribonuclease A (RNase-A) as the bio-template. The as-prepared AuNCs not only fluoresce strongly at the near-infrared (NIR) region (λ(em) = 682 nm), but also exhibit an elevated QY of â¼12%. Additionally, the RNase-A-encapsulated AuNC (RNase-A-AuNC) displays an exceptionally large Stokes shift of â¼210 nm as well as a single dominant fluorescence lifetime of â¼1.5 µs, about three orders of magnitude longer than biological autofluorescence. Furthermore, by coupling vitamin B(12) (VB(12)) to the RNase-A-AuNC, we develop a multifunctional nanoplatform that is suitable for simultaneous targeting and imaging of cancer at the cellular level using Caco-2 cell lines as an in vitro model. Since VB(12) has effective uptake pathways in the digestive system, this nanoplatform may have potential for targeted oral drug delivery in vivo.