85
Thomas Mavromoustakos et al. (eds.), Supramolecules in Drug Discovery and Drug Delivery: Methods and Protocols,
Methods in Molecular Biology, vol. 2207, https://doi.org/10.1007/978-1-0716-0920-0_7,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
Chapter 7
Multisensitive Polymeric Nanocontainers as Drug Delivery
Systems: Biological Evaluation
Maria Theodosiou, Theodora Koutsikou, and Eleni K. Efthimiadou
Abstract
This chapter focuses on the in vitro biological evaluation of multisensitive nanocontainers as drug delivery
systems for cancer treatment. Cancer tissues possess some unique characteristics such as increased temperature due to inflammation, thermal vulnerability (40–45 °C), low cellular pH, and redox instabilities. The
employment of polymers bearing pH, thermo, and/or redox sensitivities in the synthesis of hollow polymeric nanostructures has led to the formulation of a variety of drug delivery vehicles that are capable of
targeted delivery and trigger specific drug release. The cavity in the structure allows for the encapsulation
of anticancer drugs as well as other moieties with anticancer activity, like iron oxide magnetic nanoparticles. The drug loading and release capability of the nanocontainers is evaluated prior to biological studies
in order to determine the concentration of the drug in the structure. The in vitro assessment includes
cytotoxicity studies, quantitatively through the colorimetric MTT assay as well as qualitatively via the
scratch-wound healing assay, on both cancer and healthy cell lines. The cellular localization of the studied
drug-loaded and unloaded nanocontainers is determined through confocal fluorescence microscopy.
Key words Polymeric nanocontainers, Nanomedicine, MTT assay, Scratch-wound healing assay,
Multisensitive drug delivery systems, Triggered drug release
1 Introduction
Nanomedicine is the field of medicine where organic, inorganic,
and/or hybrid materials at the nanoscale (nanomaterials) can be
used in therapeutic and diagnostic (theranostic) applications for
various diseases [1]. Great focus has been given in the application
of nanomaterials as site-specific and sustainable drug delivery systems (DDS) [2, 3].
One of the main nanomaterials in DDS research are the stimuli-“sensitive” or -“responsive” polymers [4]. Polymers, depending on their physicochemical characteristics, display structural
changes upon endogenous or exogenous triggers and can be classified accordingly into three main categories: (i) pH-sensitive polymers incorporating weak acidic or basic ionic functional groups
which respond to endogenous tissue pH variations [5], (ii) ther-
Thomas Mavromoustakos et al. (eds.), Supramolecules in Drug Discovery and Drug Delivery: Methods and Protocols,
Methods in Molecular Biology, vol. 2207, https://doi.org/10.1007/978-1-0716-0920-0_7,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
Chapter 7
Multisensitive Polymeric Nanocontainers as Drug Delivery
Systems: Biological Evaluation
Maria Theodosiou, Theodora Koutsikou, and Eleni K. Efthimiadou
Abstract
This chapter focuses on the in vitro biological evaluation of multisensitive nanocontainers as drug delivery
systems for cancer treatment. Cancer tissues possess some unique characteristics such as increased temperature due to inflammation, thermal vulnerability (40–45 °C), low cellular pH, and redox instabilities. The
employment of polymers bearing pH, thermo, and/or redox sensitivities in the synthesis of hollow polymeric nanostructures has led to the formulation of a variety of drug delivery vehicles that are capable of
targeted delivery and trigger specific drug release. The cavity in the structure allows for the encapsulation
of anticancer drugs as well as other moieties with anticancer activity, like iron oxide magnetic nanoparticles. The drug loading and release capability of the nanocontainers is evaluated prior to biological studies
in order to determine the concentration of the drug in the structure. The in vitro assessment includes
cytotoxicity studies, quantitatively through the colorimetric MTT assay as well as qualitatively via the
scratch-wound healing assay, on both cancer and healthy cell lines. The cellular localization of the studied
drug-loaded and unloaded nanocontainers is determined through confocal fluorescence microscopy.
Key words Polymeric nanocontainers, Nanomedicine, MTT assay, Scratch-wound healing assay,
Multisensitive drug delivery systems, Triggered drug release
1 Introduction
Nanomedicine is the field of medicine where organic, inorganic,
and/or hybrid materials at the nanoscale (nanomaterials) can be
used in therapeutic and diagnostic (theranostic) applications for
various diseases [1]. Great focus has been given in the application
of nanomaterials as site-specific and sustainable drug delivery systems (DDS) [2, 3].
One of the main nanomaterials in DDS research are the stimuli-“sensitive” or -“responsive” polymers [4]. Polymers, depending on their physicochemical characteristics, display structural
changes upon endogenous or exogenous triggers and can be classified accordingly into three main categories: (i) pH-sensitive polymers incorporating weak acidic or basic ionic functional groups
which respond to endogenous tissue pH variations [5], (ii) ther-