Arunkumar Pitchaimani

Engineered biomimetic nanoabsorbent for cellular detoxification of chemotherapeutics

An approach to reduce the nonspecific cytotoxicity of chemotherapeutics has been put-forth using a biomimetic nanoabsorbent (NAb) as a detoxifying agent. The engineered NAb possesses a tunable drug absorption ability depending on the charge of the absorbing molecule, in which the cooperative absorption ability of the core and shell significantly reduces the cellular toxicity.

Impact of cell adhesion and migration on nanoparticle uptake and cellular toxicity

In vitro cell-nanoparticle (NP) studies involve exposure of NPs onto the monolayer cells growing at the bottom of a culture plate, and assumed that the NPs evenly distributed for a dose-responsive effect. However, only a few proportion of the administered dose reaches the cells depending on their size, shape, surface, and density. Often the amount incubated (administered dose) is misled as a responsive dose. Herein, we proposed a cell adhesion-migration (CAM) strategy, where cells incubated with the NP coated cell culture substrate to maximize the cell-NP interaction and investigated the physiological properties of the cells. In the present study, cell adhesion and migration pattern of human breast cancer cell (MCF-7) and mouse melanoma cell (B16-F10) on cell culture substrate decorated with toxic (cetyltrimethylammonium bromide, CTAB) and biocompatible (poly (sodium 4-styrenesulphonate), PSS) gold nanoparticles (AuNPs) of different sizes (5 and 40nm) were investigated and evaluated for cellular uptake efficiency, proliferation, and toxicity. Results showed enhanced cell adhesion, migration, and nanoparticle uptake only on biocompatible PSS coated AuNP, irrespective of its size. Whereas, cytotoxic NP shows retard proliferation with reduced cellular uptake efficiency. Considering the importance of cell adhesion and migration on cellular uptake and cytotoxicity assessment of nanoparticle, CAM strategy would hold great promises in cell-NP interaction studies.

Synthesis and Characterization of Biomimetic Hydroxyapatite Nanoconstruct Using Chemical Gradient across Lipid Bilayer

In this study, we synthesized biomimetic hydroxyapatite nanoconstruct (nanosized hydroxyapatite, NHAp) using a double emulsion technique combined with a chemical gradient across a lipid bilayer for surface modification of a titanium (Ti) implant. The synthesized NHAp was characterized by dynamic light scattering, X-ray diffraction, transmission electron microscopy, and Fourier transform infrared (FTIR) spectroscopy, and it was further tested for its biocompatibility and in vitro proliferation efficacy using normal human osteoblasts (NHOst). The results showed that the synthesized NHAp had a hydrodynamic diameter of ∼200 nm with high aqueous stability. The chemistry of the NHAp was confirmed by FTIR spectroscopic analysis. Typical FTIR vibrational bands corresponding to the phosphate group (PO4(3-)) present in hydroxyapatite (HAp) were observed at 670, 960, and 1000 cm(-1). A broad band at 3500 cm(-1) confirmed the presence of a structural -OH group in the NHAp. Powder X-ray crystallographic diffraction further confirmed the formation of NHAp with characteristic reflections in (002), (211), (130), and (213) planes at respective 2θ degrees. These reflection planes are similar to those of typical HAp crystallized toward (002) and (211) crystallographic planes. The mechanism of the formation of NHAp was studied using the fluorescence resonance energy transfer (FRET) technique. The FRET study showed the fluorescent recovery of a donor fluorophore and the mechanism of the insertion of lipids into nanodroplets obtained from the first water-in-oil (w/o) emulsion during the formation of the second oil-in-water (o/w) emulsion. With these confirmations, we further studied NHOst cell proliferation on a Ti surface. When NHOst were cultured on the Ti surface coated with the NHAp, a distinct proliferation pattern and cell-cell communication via cytoplasmic extension on the substrate surface were observed. In contrast, a bare Ti surface showed diminished cell size with minimal adherence. This result indicates that our NHAp covered with a phospholipid bilayer provides a proper environment essential for cell adhesion, which is especially important for bone implants, and the inclusion of NHAp on the Ti substrate would be an effective support for long-term sustainability of implants.

Engineered Nanomedicine with Alendronic Acid Corona Improves Targeting to Osteosarcoma

We engineered nanomedicine with the stealth corona made up of densely packed bone seeking ligand, alendronic acid. In a typical nanoconstruct, alendronic acid is conjugated with hydrophilic head moiety of phospholipid that has an ability to self-assemble with hydrophobic polymeric core through its hydrophobic long carbon-chain. Proposed nanomedicine has three distinct compartments namely; poly(l-lactic-co-glycolic acid) polymeric core acting as a drug reservoir and skeleton of the nanoconstruct, phospholipid monolayer covers the core acting as a diffusion barrier, and a densely packed alendronic acid corona acting as a stabilizer and targeting moiety. Thus engineered nanomedicine attain spherical entity with ~90 ± 6 nm having negative zeta potential, −37.7 ± 2 mV, and has an ability to load 7 ± 0.3 wt% of doxorubicin. In-vitro bone targeting efficiency of nanomedicine was studied using hydroxyapatite crystals as a bone model, and found significant accumulation of nanoparticle in the crystals. Moreover, cellular internalization studies with mouse osteosarcoma confirm the selectivity of nanomedicine when compared to its internalization in non-targeted mouse melanoma. This nanomedicine shows prolong stability in serum and deliver the drug into the cell exhibiting an IC50 of 3.7 μM. Given the strong interacting property of alendronic acid with bone, the proposed nanomedicine hold promises in delivering drug to bone microenvironment.

Design and characterization of gadolinium infused theranostic liposomes

Multifunctional theranostic gadolinium infused liposomes containing the chemotherapeutic drug, doxorubicin (DOX), in its core are designed as potential candidates for diagnosis and therapy of various cancers. In these theranostic liposomes, Gd3+ ions are chelated by the macrocyclic chelator 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, which is covalently linked to the hydrophilic head moiety of the phospholipid. Therefore, the Gd3+ labels are an integral part of the lipid bilayer of liposomal construct. The physical properties of the Gd-infused liposomes were characterized including drug loading, drug release kinetics, cellular consequences, and T1 weighted magnetic properties. The theranostic liposomes exhibits uniform size distribution (hydrodynamic size of 150 ± 10 nm) and a high drug loading efficiency with sustained drug release characteristic. Due to stable Gd3+ in their lipid bilayers, the theranostic liposomes displayed an enhanced r1 relaxivity of 12.3 mM−1 s−1 at 14.1 T, which is three times higher than that of currently used Gd-based contrast agent (Magnivest®: 4.1 mM−1 s−1, 1.4 T). Moreover, cellular uptake studies revealed that these theranostic liposomes exhibit higher structural integrities inside B16-F10 melanoma cells. It shows uniform intracellular distribution throughout the cells. In vitro cytotoxicity revealed that Gd-infused liposomes have excellent biocompatibility without significant cytotoxicity, whereas they showed higher cellular toxicity when loaded with DOX against B16-F10. The observed toxicity was equivalent to the same concentration of free DOX. With regard to the ongoing development in theranostic water-dispersible nanoformulations, the current formulation where Gd3+ ions are encapsulated by a macrocyclic chelator and covalently tethered to the hydrophilic head moiety of a lipid, is rather promising and holds great promise.