A semiautomated microfluidic platform for real-time investigation of nanoparticles' cellular uptake and cancer cells' tracking

Aims: develop a platform composed of labeled dendrimer nanoparticles and a microfluidic device for real-time monitoring of cancer cells fate. Materials and Methods: The physicochemical and biological characterization of the developed Carboxymethyl-chitosan/poly(amidoamine) (CMCht/PAMAM) dendrimer nanoparticles were performed using TEM, AFM, Zeta Sizer, DSC and cytotoxicity screening. Cancer cell lines derived from different tumor types, including HeLa (Cervical Carcinoma), HCT-116 (Colon Carcinoma) and U87MG (Glioblastoma), were exposed to different concentrations of CMCht/PAMAM dendrimer nanoparticles over a period of 3 days (MTS/DNA). Results: Nanoparticles were successfully modified with an average size of 50 nm. Internalization levels go from 87% to 100% in static and from 95% to 100% in dynamic conditions. Viability levels range from 95% to 100% in static and from 90% to 100% in dynamic conditions, being HCT the most sensitive to the presence of the NP. Conclusions: the results show different responses to the presence of 0.5 mg.mL-1 dendrimer nanoparticles when comparing static to dynamic conditions, with a tendency towards higher sensitivity when subjected to confinement. This work demonstrated that the proposed microfluidic-based platform allows real-time cell monitoring, which, upon more studies, namely the assessment of the drug release effect, could be used for cancer theranostics.FR Maia acknowledges ERC-2012-ADG 20120216–321266 (ComplexiTE) for her Postdoc scholarship. JM Oliveira thanks Portuguese Foundation for Science and Technology (FCT) for his distinction attributed under the FCT Investigator program (IF/00423/2012). BM Costa also thanks Portuguese Foundation for Science and Technology (PTDC/SAU-GMG/113795/2009 and IF/00601/2012 to BM Costa), Fundação Calouste Gulbenkian (BM Costa) and Liga Portuguesa Contra o Cancro (BM Costa). MR Carvalho also thanks the funding through the LA ICVS/3Bs project (UID/Multi/50026/2013). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.