Abstract: Ultrasound (US) is widely used in medical field for
a variety diagnostic techniques but, in recent years, it has also
been creating great interest for therapeutic aims. Regarding drug
delivery, the use of US as an activation source provides better
spatial delivery confinement and limits the undesired side effects.
However, at present there is no complete characterization
at a fundamental level of the different signals produced by
sono-activated nanocarriers. Therefore, the aim of this study
is to obtain a metrological characterization of the cavitation
phenomena induced by US through three parallel investigation
approaches. US was focused into a channel of a customized
phantom in which a solution with oxygen-loaded nanodroplets
(OLNDs) was led to flow and the cavitation activity was
monitored. Both quantitative and qualitative real-time analysis
were performed giving information about the dynamics of
bubble formation, oscillation and final implosion with respect
to the working acoustic pressure and the type of nanodroplets,
compared with pure water. From this analysis a possible
interpretation of the observed results is proposed.
Abstract: Early detection of cancer could save human life and quality in insidious cases by advanced biomedical imaging techniques. Designing targeted detection system is necessary in order to protect of healthy cells. Electrospun nanofibers are efficient and targetable nanocarriers which have important properties such as nanometric diameter, mechanical properties, elasticity, porosity and surface area to volume ratio. In the present study, indocyanine green (ICG) organic dye was stabilized and encapsulated in polymer matrix which polyethylene oxide (PEO) and chitosan (CHI) multilayer nanofibers via co-axial electrospinning method at one step. The co-axial electrospun nanofibers were characterized as morphological (SEM), molecular (FT-IR), and entrapment efficiency of Indocyanine Green (ICG) (confocal imaging). Controlled release profile of PEO/CHI/ICG nanofiber was also evaluated up to 40 hours.
Abstract: In this study, we developed and simulated nano-drug delivery systems efficacy in compare to free drug prescription. Computational models can be utilized to accelerate experimental steps and control the experiments high cost. Molecular dynamics simulation (MDS), in particular NAMD was utilized to better understand the anti-cancer drug interaction with cell membrane model. Paclitaxel (PTX) and dipalmitoylphosphatidylcholine (DPPC) were selected for the drug molecule and as a natural phospholipid nanocarrier, respectively. This work focused on two important interaction parameters between molecules in terms of center of mass (COM) and van der Waals interaction energy. Furthermore, we compared the simulation results of the PTX interaction with the cell membrane and the interaction of DPPC as a nanocarrier loaded by the drug with the cell membrane. The molecular dynamic analysis resulted in low energy between the nanocarrier and the cell membrane as well as significant decrease of COM amount in the nanocarrier and the cell membrane system during the interaction. Thus, the drug vehicle showed notably better interaction with the cell membrane in compared to free drug interaction with the cell membrane.
Abstract: Drug delivery to target human acute myeloid leukemia (AML) using a nanoparticulate chemotherapeutic formulation that can deliver drugs selectively to AML cancer is hugely needed. In this work, we report the development of a nanoformulation made of polymeric-stabilized multifunctional magnetic iron oxide nanoparticles (PMNP) loaded with the anticancer drug Doxorubicin (Dox) as a promising drug carrier to treat AML. Dox@PMNP conjugates simultaneously exhibited high drug content, maximized fluorescence, and excellent release properties. Nanoparticulate uptake and cell death following addition of Dox@PMNPs were then evaluated in different types of human AML target cells, as well as on normal human cells. While the unloaded MNPs were not toxic to any of the cells, Dox@PMNPs were found to be highly toxic to the different AML cell lines, albeit at different inhibitory concentrations (IC50 values), but showed very little toxicity towards the normal cells. In comparison, free Dox showed significant potency concurrently to all the cell lines, suggesting huge potentials for the use of Dox@PMNPs as selective AML anticancer cargos. Live confocal imaging, fluorescence and electron microscopy confirmed that Dox is indeed delivered to the nucleus in relatively short periods of time, causing apoptotic cell death. Importantly, this targeted payload may potentially enhance the effectiveness of the drug in AML patients and may further allow physicians to image leukemic cells exposed to Dox@PMNPs using MRI.
Abstract: Various nanomaterials can be used as a drug delivery
vehicles in nanomedicine, called nanocarriers. They can either be
organic or inorganic, synthetic or natural-based. Although synthetic
nanocarriers are easier to produce, they can often be toxic for the
organism and thus not suitable for use in treatment. From naturalbased
nanocarriers, the most commonly used are protein cages or
viral capsids. In this work, virus bacteriophage λ was used for
delivery of different cytotoxic drugs (cisplatin, carboplatin,
oxaliplatin and doxorubicin). Large quantities of phage λ were
obtained from phage λ-producing strain of E. coli cultivated in
medium with 0.2% maltose. After killing of E. coli with chloroform
and its removal by centrifugation, the phage was concentrated by
ultracentrifugation at 130 000×g and 4°C for 3 h. The encapsulation
of the drugs was performed by infusion method and four different
concentrations of the drugs were encapsulated (200; 100; 50; 25
μg·mL-1). Free drug molecules were removed by filtration. The
encapsulation was verified using the absorbance for doxorubicin and
atomic absorption spectrometry for platinum cytostatics. The amount
of encapsulated drug linearly increased with the increasing
concentration of applied drug with the determination coefficient
R2=0.989 for doxorubicin; R2=0.967 for cisplatin; R2=0.989 for
carboplatin and R2=0.996 for oxaliplatin. The overall encapsulation
efficiency was calculated as 50% for doxorubicin; 8% for cisplatin;
6% for carboplatin and 10% for oxaliplatin.
Abstract: To ensure targeting of apoferritin nanocarrier with
encapsulated doxorubicin drug, we used a peptide linker based on a
protein G with N-terminus affinity towards Fc region of antibodies.
To connect the peptide to the surface of apoferritin, the C-terminus of
peptide was made of cysteine with affinity to gold. The surface of
apoferritin with encapsulated doxorubicin (APODOX) was coated
either with gold nanoparticles (APODOX-Nano) or gold(III) chloride
hydrate reduced with sodium borohydride (APODOX-HAu). The
reduction with sodium borohydride caused a loss of doxorubicin
fluorescent properties and probably accompanied with the loss of its
biological activity. Fluorescent properties of APODOX-Nano were
similar to the unmodified APODOX; therefore it was more suited for
the intended use. To evaluate the specificity of apoferritin modified
with antibodies, ELISA-like method was used with the surface of
microtitration plate wells coated by the antigen (goat anti-human IgG
antibodies). To these wells, the nanocarrier was applied. APODOX
without the modification showed 5× lower affinity to the antigen than
APODOX-Nano modified gold and targeting antibodies (human IgG
antibodies).
Abstract: Aim of this work was to compare the efficacy of two
loading methods of proteins onto polymeric nanocarriers: adsorption
and encapsulation methods. Preliminary studies of protein loading
were done using Bovine Serum Albumin (BSA) as model protein.
Nanocarriers were prepared starting from polylactic co-glycolic acid
(PLGA) polymer; production methods used are two different variants
of emulsion evaporation method. Nanoparticles obtained were
analyzed in terms of dimensions by Dynamic Light Scattering and
Loading Efficiency of BSA by Bradford Assay. Loaded
nanoparticles were then submitted to in-vitro protein dissolution test
in order to study the effect of the delivery system on the release rate
of the protein.
Abstract: Identifying the nature of protein-nanoparticle
interactions and favored binding sites is an important issue in
functional characterization of biomolecules and their physiological
responses. Herein, interaction of silver nanoparticles with lysozyme
as a model protein has been monitored via fluorescence spectroscopy.
Formation of complex between the biomolecule and silver
nanoparticles (AgNPs) induced a steady state reduction in the
fluorescence intensity of protein at different concentrations of
nanoparticles. Tryptophan fluorescence quenching spectra suggested
that silver nanoparticles act as a foreign quencher, approaching the
protein via this residue. Analysis of the Stern-Volmer plot showed
quenching constant of 3.73 μM−1. Moreover, a single binding site in
lysozyme is suggested to play role during interaction with AgNPs,
having low affinity of binding compared to gold nanoparticles.
Unfolding studies of lysozyme showed that complex of lysozyme-
AgNPs has not undergone structural perturbations compared to the
bare protein. Results of this effort will pave the way for utilization of
sensitive spectroscopic techniques for rational design of
nanobiomaterials in biomedical applications.