Rapid Green Synthesis of Biogenic Silver Nanoparticles Using Cinnamomum tamala Leaf Extract and its Potential Antimicrobial Application Against Clinically Isolated Multidrug-Resistant Bacterial Strains.

Cinnamomum tamala is Indian bay leaves also known as Tej patta commonly used in the preparation of delicious food for its sweet aroma and tremendous medicinal values. In this study, the significant concentration-dependent free radical scavenging and antioxidant efficacy of the aqueous extracts of bay leaves has been determined using DPPH (2, 2-diphenyl-l-picrylhydrazyl) radical scavenging, ferric ion-reducing power assay, and hydrogen peroxide radical scavenging assay. The leaf extract has also been utilized in the rapid synthesis of silver nanoparticles (AgNPs) under mild conditions (30 min reaction time at 70 °C) without the addition of extra stabilizing or capping agents. Mostly spherical shaped particles were formed with diameter ranging from 10 to 12 nm as evident by HRTEM imaging. The silver nanoparticles were also characterized using FTIR, XRD, and UV-visible spectroscopic techniques. The antibacterial effect of the synthesized AgNPs was studied against three clinically isolated multidrug-resistant bacterial strains (Escherichia coli (EC-1), Klebsiella pneumonia (KP-1), and Staphylococcus aureus (SA-1)). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of AgNPs against EC-1 were 12.5 and 15 µg/mL and in SA-1 were 10 and 50 µg/mL, and in the case of KP-1, both values were 12.5 µg/mL. It was also noted that 8 h treatment duration using AgNPs was sufficient to eliminate all types of bacterial growth as evidenced by time-dependent killing kinetic assays. The biocompatibilities of AgNPs were also tested against human health RBCs, and it was observed that it did not show any significant toxicity up to 50 µg/mL concentration.

Hierarchical Self-Assembly of a Renewable Nanosized Pentacyclic Dihydroxy-triterpenoid Betulin Yielding Flower-Like Architectures.

Betulin, a naturally occurring 6-6-6-6-5 pentacyclic dihydroxy-triterpenoid, is extractable from the bark of white birch (Betula papyrifera). We report the first self-assembly properties of betulin in different liquids. The molecule spontaneously self-assembled in different media, yielding flower-like architectures of nano- to micrometers diameters via the formation of fibrillar networks. The self-assemblies have been characterized by optical microscopy, scanning electron microscopy, transmission electron microscopy, dynamic light scattering, FTIR, and X-ray diffraction studies. The porous microstructure of the self-assemblies has been utilized for the entrapment of fluorophore such as rhodamine-B and the anticancer drug doxorubicin. Moreover, the removal of toxic dyes such as rhodamine 6G, crystal violet, methylene blue, and cresol red has also been demonstrated.

Self assembled nano fibers of betulinic acid: A selective inducer for ROS/TNF-alpha pathway mediated leukemic cell death.

The main complication in betulinic acid (BA) based drug delivery has been observed due to its bulk structure. The present study demonstrates the potential effects of self assembled nano size betulinic acid (SA-BA) by treating human leukemic cell lines (KG-1A and K562) and its normal counterpart. Self assembly property of BA was investigated using SEM and DLS study which showed that the BA forms fibrous structure having few nanometers in diameter. Selective anti-leukemic efficacy study of SA-BA was investigated by cell viability assay. Mode of leukemic cell death and probable pathway of apoptosis was monitored by measuring ROS level, pro and anti-inflammatory cytokine status and expression of caspase-8 and caspage-3 by immunocytochemistry. Higher efficacy of SA-BA over non-assemble BA was monitored toward cancer cells, with no relevant toxicity to normal blood cells. SA-BA facilitated reactive oxygen species (ROS) mediated leukemic cell death, confirmed by pre-treatment of N-acetyl-L-cysteine. Induction of apoptosis by SA-BA treatment increased pro-inflammatory cytokines, specifically potentiated TNF-α mediated cell death, confirmed by expression of caspase-8 and caspage-3 by immunocytochemistry. This study explored the better anti-leukemic efficacy of SA-BA over BA and this modification will enrich the use of BA in cancer therapy.

Self-assembled betulinic acid augments immunomodulatory activity associates with IgG response.

Studies relating to the adjuvanic role of self assembly, nanosized betulinic acid (SA-BA) are relatively limited. The concept of immunostimulatory activity of SA-BA is based on the activation of immune system against cancer antigen. This study showed that SA-BA, a pentacyclic triterpene isolated from the bark of the Ziziphus jujube tree, elevated the immunological functions of cancer antigen in anticancer immunotherapy. We found that, SA-BA pulsed human macrophages secreted elevated level of pro-inflammatory cytokines with an increased CD4(+) cell population. Pulse macrophages were also significantly arrested the KG-1A and K562 cell growth in vitro setup at 1:10 ratio for 48h. The use of TNF-α inhibitors confirmed the association between SA-BA with TNF-α function. SA-BA pulsed macrophages displayed substantial T cell allostimulatory capacity and promoted the generation of cytotoxic T lymphocytes (CTLs). The adjuvanticity of SA-BA was proved by the generation of in vivo IgG response. Collectively, these findings will enrich the biomedical applications of SA-BA as a potent immune stimulating agent. Moreover, the macrophage stimulating efficacy of SA-BA might be an effective way in the cancer immunotherapy.

Self-assembled betulinic acid protects doxorubicin induced apoptosis followed by reduction of ROS-TNF-α-caspase-3 activity.

Doxorubicin (DOX) is a well-known drug used to treat a wide range of solid tumor and hematological malignancies, but the use of this drug is now restricted owing to its severe side effects, including normal cellular toxicity. This study was conducted to evaluate the potency of self-assembled betulinic acid (SA-BA) against DOX induced chemotherapeutic toxicity in human peripheral blood lymphocytes (PBLs). The isolated betulinic acid from the bark of Ziziphus jujuba tree was purified by column chromatography and characterized by FT-IR, XRD, (1)H NMR and self-assembly property was investigated by SEM imaging. DOX treatment produced significant reduction of viability of PBLs mainly by lowering cellular anti-oxidant pool and elevating the reactive oxygen species level. Pre-treatment with SA-BA followed by DOX exposure for 24h protected the PBLs from DOX induced oxidative stress. Potent anti-apoptotic role of SA-BA was also confirmed by FACS analysis and western blot assay. Severe inflammation is one of the major concerns in DOX treatment. We found that pre-treatment with SA-BA on PBLs significantly protected the PBLs from DOX induced inflammation. Thus, our finding confirms that SA-BA can be used to ameliorate the cytotoxic effects of DOX, which can be a helpful strategy during DOX mediated chemotherapy in cancer patients.

First self-assembly study of betulinic acid, a renewable nano-sized, 6-6-6-6-5 pentacyclic monohydroxy triterpenic acid.

Self-assembly of betulinic acid, a renewable nano-sized 6-6-6-6-5 pentacyclic triterpenic acid studied in twenty two organic liquids and alcohol-water mixtures showed that it self-assembled in all the liquids studied affording strong gels in nineteen organic liquids and also in alcohol-water mixtures. Optical and electron microscopy and atomic force microscopy studies revealed fibrillar networks having fibers of nano- to micro-metre cross sections and micrometre lengths.