Biology subjects

Kansara, K.

Publications and source records attributed to Kansara, K..

9 recordsLinked to original sources

bioRxiv2024

Neurotransmitter loaded DNA nanocages as potential therapeutics for α-synuclein based neuropathies in cells and in vivo

Parkinsons disease is one of the neuropathies characterized by accumulation of -synuclein protein, leading to motor dysfunction. Levodopa is the gold standard treatment, however, in long term usage, it leads to levodopa induced dyskinesia (LID). New therapeutic options are need of the hour to treat the -synuclein based neuropathies. The role of imbalance of neurotransmitters other than dopamine has been underestimated in -synuclein based neuropathies. Here, we explore the role of serotonin, epinephrine and norepinephrine as a therapeutic moiety. For the efficient in vivo delivery, we use DNA nanotechnology-based DNA tetrahedra that has shown the potential to cross the biological barriers. In this study, we explore the use of DNA nanodevices, particularly DNA tetrahedron functionalized with neurotransmitters, as a novel therapeutic approach for MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) induced Parkinsons disease in PC12 cellular system. We first establish the effect of these nanodevices on clearance of -synuclein protein in cells. We follow the study by understanding the various cellular processes like ROS, iron accumulation and lipid peroxidation. We also explore the effect of the neurotransmitter loaded nanodevices in in vivo zebrafish model. We show that neurotransmitter loaded DNA nanocages can potentially clear the MPTP induced -synuclein aggregates in cells and in vivo. The findings of these work open up new avenues for use of DNA nanotechnology by functionalizing it with neurotransmitters for future therapeutics in treatment of neurodegenerative diseases such as Parkinsons disease. TOC O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=131 SRC="FIGDIR/small/626934v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@94c347org.highwire.dtl.DTLVardef@a08756org.highwire.dtl.DTLVardef@1153704org.highwire.dtl.DTLVardef@1cefebb_HPS_FORMAT_FIGEXP M_FIG C_FIG TD:NT can clear -synuclein by targeting the ferroptosis pathway.

bioengineering↗

bioRxiv2024

Peptide modified, programmable DNA tetrahedra to modulate autophagy in biological systems

Autophagy is a critical cellular pathway for degrading and recycling damaged components, essential for maintaining cellular homeostasis. Dysregulation of autophagy contributes to various diseases, including neurodegenerative disorders, cancers, and metabolic syndromes, highlighting the therapeutic potential of controlled autophagy induction. However, current autophagy inducers often lack specificity and may inadvertently trigger apoptosis, limiting their clinical utility. Here, we present a DNA tetrahedron-BH3 peptide nanosystem (Tdpep) engineered to selectively induce autophagy by disrupting the Beclin 1-Bcl2 interaction, a pivotal regulatory point in autophagy initiation. Tdpep, functionalized with a BH3 peptide targeting Bcl2, demonstrated efficient cellular uptake and minimal cytotoxicity in HeLa cells at concentrations up to 200nM. Autophagy induction was confirmed by increased LC3B puncta formation and fluorescence intensity comparable to that induced by rapamycin. Autophagy flux analysis of Tdpep with bafilomycin A1 validated enhanced autophagic activity rather than flux inhibition. Furthermore, Tdpep treatment significantly reduced cellular ROS levels, indicating effective autophagic turnover. Apoptosis assays showed that Tdpep did not induce apoptosis, confirming its selective autophagy induction. Furthermore, Tdpep nanosystem also induced autophagy in Danio rerio larvae in vivo model. Thus, this targeted DNA tetrahedron nanosystem provides a precise autophagy modulation platform with minimized off-target effects, offering a promising therapeutic strategy for diseases associated with autophagy dysfunction. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/621781v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@193cde5org.highwire.dtl.DTLVardef@b2a041org.highwire.dtl.DTLVardef@13711eaorg.highwire.dtl.DTLVardef@792bb5_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2023

DNA Tetrahedral Nanocages as a Promising Nanocarrier for Dopamine Delivery in Neurological Disorders

Dopamine is a neurotransmitter in the central nervous system that is essential for many bodily and mental processes, and a lack of it can cause Parkinsons disease. DNA tetrahedral (TD) nanocages are promising in bio-nanotechnology, especially as a nanocarrier. TD is highly programmable, biocompatible, and capable of cell differentiation and proliferation. It also has tissue and blood-brain barrier permeability, making it a powerful tool that could overcome potential barriers in treating neurological disorders. In this study, we used DNA-TD as a carrier for Dopamine to cells and zebrafish embryos. We investigated the mechanism of complexation between TD and dopamine hydrochloride using gel electrophoresis, fluorescence and circular dichroism (CD) spectroscopy, atomic force microscopy (AFM), and molecular dynamic (MD) simulation tools. Further, we demonstrate these Dopamine-loaded DNA tetrahedral nanostructures cellular uptake and differentiation ability in SH-SY5Y neuroblastoma cells. Furthermore, we extended the study to zebrafish embryos as a model organism to examine survival and uptake. The research provides valuable insights into the complexation mechanism and cellular uptake of dopamine-loaded DNA tetrahedral nanostructures, paving the way for further advancements in nanomedicine for Parkinsons disease and other neurological disorders.

bioengineering↗

bioRxiv2023

DNA tetrahedron as a carrier of doxorubicin for metastatic breast cancer treatment

Metastatic breast cancer is a significant clinical challenge calling for novel and efficient therapeutic approaches. DNA tetrahedron, a highly programmable nanocage, offers some promising attributes including biocompatibility, stability, and functionalization making it an attractive candidate for drug delivery. In this study, we have explored the potential of DNA tetrahedron as a carrier of doxorubicin, a DNA and RNA synthesis-inhibiting chemotherapy drug. We have encapsulated doxorubicin in DNA tetrahedron (TD: Dox) and subsequently focused on metastatic breast cancer cells for the effect of the same. We showed that TD: Dox has the potential to inhibit the migration of cancerous cells in the 2D model and inhibit the invasion of tumor cells in the 3D model as well. This system also can be uptaken in in vivo zebrafish model as well. Overall, this study promises the TD: Dox system as an ideal drug delivery model and a viable approach for metastatic breast cancer treatment.

bioengineering↗

bioRxiv2023

Dopamine functionalized, red carbon quantum dots for in vivo bioimaging, cancer therapeutics, and neuronal differentiation

One of the crucial requirements of quantum dots for biological applications is their surface modifications for very specific and enhanced biological recognition and uptake. Toward this, we present the green synthesis of bright, red-emitting carbon quantum dots derived from mango leaf extract (mQDs). These mQDs are conjugated electrostatically with dopamine to form mQDs-dopamine (mQDs: DOPA) bioconjugates. Bright red fluorescence of mQDs was used for bioimaging and uptake in multiple cell lines, tissues, and in vivo models like zebrafish. mQDs exhibited the highest uptake in brain tissue as compared to others. mQD:DOPA conjugate induced cellular toxicity only in cancer cells while showing increased uptake in epithelial cells and zebrafish. Additionally, the mQDs: DOPA promoted neuronal differentiation of SH-SY5Y cells to complete neurons. Both mQDs and mQDs: DOPA exhibited potential for higher collective cell migrations implicating their future potential as next-generation tools for advanced biological and biomedical applications. TOCmQDs were electrostatically conjugated with dopamine (DOPA) to form the mQDs: DOPA bioconjugate. mQDs are used to image cells, tissues, and zebrafish embryos. mQDs: DOPA kills cancer cells, differentiates neuronal cells, and increases the uptake of mQDs in zebrafish embryos. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/545347v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@f521a4org.highwire.dtl.DTLVardef@173a8f0org.highwire.dtl.DTLVardef@585271org.highwire.dtl.DTLVardef@8c959d_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2023

Red Fluorescent Carbon Nanoparticles Derived from Spinacia oleracea L.: A Versatile Tool for Bioimaging & Biomedical Applications

Carbon-based fluorescent quantum dots are an emerging class of nanoparticles for targeted bioimaging and biomedical applications. We present a facile microwave-assisted approach for synthesizing carbon nanoparticles with bright red fluorescence using ethanolic extracts of Spinacia oleracea leaves, with a quantum yield of 94.67%. These nanoparticles, called CNPs, ranging from 15-50 nm, demonstrated fluorescence emission in the near-infrared (NIR) region between 650 and 700 nm, independent of excitation wavelength. Upon excitation at a wavelength of 410 nm, they exhibit an emission maxima peak at 672 nm. The significant uptake of CNPs in mammalian cells and zebrafish larvae highlights their potential as bioimaging agents in diverse biomedical applications in vivo. Further, these quantum dots enhance cellular proliferation and migration as observed by wound healing assay in mammalian cells, indicating their possible application in tissue engineering and regenerative medicine. These findings suggest that biosynthesized carbon nanoparticles possess significant potential for biomedical activities, which can serve as a robust benchmark for researchers towards promoting sustainability.

bioengineering↗

bioRxiv2023

Lipid modification of DNA nanocages enhances cellular uptake, migration, and in vivo uptake

The extraordinary self-assembling nature of DNA nanostructures and high functionality enables the formulation of DNA nanostructures with multiple chemical and biological molecules. How the whole organisms in native as well as modified; and how stable they are inside take up exactly these modified DNA nanostructures the organisms still remains to be explored. Here we report the fabrication and evaluation of a new conjugate of a cationic lipid, N-[one-(two, 3-dioleyloxy) propyl]-N, N, N-trimethylammonium chloride (DOTMA) and DNA tetrahedron nanostructure (TdN) for the enhanced uptake, stability, bioimaging, and biotherapeutics in cells and zebrafish (Danio rerio) eleuthero embryos as a model organism. We summarise the enhanced uptake potential of TdN-DOTMA conjugate for futuristic biomedical applications such as drug delivery, bioimaging, biosensing, and therapeutics.

bioengineering↗

bioRxiv2022

Novel class of yellow emitting carbon dots stimulate collective cell migration and 3D uptake in vivo

We present a new class of nitrogen-doped yellow fluorescent carbon dots, synthesized using a one-step hydrothermal method. These bright fluorescent nanoparticles have excitation and emission spectra near the red region of the visible light spectrum that are quite useful for bioimaging applications. Using organic molecules like ortho- phenylenediamine (OPDA), L-ascorbic acid and urea, yellow fluorescent carbon dots (CDs) were synthesized. We obtained a scalable number of CDs having an average size of 3 nm. The CDs show significant emission spectra in the yellow fluorescence region ({lambda}em= 557 nm). The CDs show remarkable stability in their fluorescence in different pH conditions, ionic stability, photostability as well as thermal stability. These CDs are efficiently uptaken by mammalian cells through clathrin-mediated pathway. Apart from in vitro studies we have also used zebrafish larvae as a 3D in vivo model, and showed that CDs were uptaken efficiently by larvae showing maximum accumulation and fluorescence in the yolk sac region and the notochord region. The CDs also offer enhancement in cell proliferation, hence showing the application in wound healing. The fluorescence of CDs is quite robust and is not affected by most external stimuli, hence can be explored as a promising bioimaging tool for targeted bioimaging and biomedical applications.

bioengineering↗

bioRxiv2022

Green emitting carbon quantum dots (GCQDs) to probe endocytic pathways in cells; for tissue and in vivo bioimaging

Small sized, carbon-based organic nanoparticles have recently gained attention due their advantage of biocompatibility, photostability and biological non-toxicity as compared to their inorganic counterparts. Herein, a new class of small (5-8 nm), green emitting fluorescent carbon quantum dots (GCQDs) were synthesized using organic substrates like citric acid and ascorbic acid in aqueous solvent containing water and ethanol. The very small size and bright green photoluminescence prompted their use for both in vitro and in vivo bioimaging. GCQDs were uptaken via clathrin mediated pathways in mouse kidney and liver primary cells. Similarly, they showed active uptake and distribution in the zebrafish embryo model system. The optical tunability and surface modification properties of these GCQDs provide a platform to be explored for them to emerge as a new class of targeted bioimaging entities, as well as tools for biomedical applications.

bioengineering↗