Publications by authors named "Chitta Ranjan Patra"

82 Publications

Biologically synthesized gold nanoparticles as a near-infrared-based bioimaging agent.

Nanomedicine (Lond) 2021 04 4;16(8):613-616. Epub 2021 Mar 4.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad, Telangana 500007, India.

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http://dx.doi.org/10.2217/nnm-2021-0027DOI Listing
April 2021

Therapeutic angiogenesis using zinc oxide nanoflowers for the treatment of hind limb ischemia in rat model.

Biomed Mater 2021 Mar 3. Epub 2021 Mar 3.

Biomaterials Group, LST Division, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, AP, Hyderabad, Andhra Pradesh, 500007, INDIA.

Critical limb ischemia (CLI) is considered as a severe type of peripheral artery diseases (PADs) which occurs due to the inadequate supply of blood to the limb extremities. CLI patients often suffer from extreme cramping pain, impaired wound healing, immobility, cardiovascular complications, amputation of the affected limb and even death. The conventional therapy for the treatment of CLI includes surgical revascularization as well as restoring angiogenesis using growth factor therapy. However, surgical revascularization is suitable for only a minor percentage of CLI patients and it is associated with high perioperative mortality rate. The use of growth factors is also limited in terms of their poor therapeutic angiogenesis potential as observed by the earlier clinical studies, which could be attributed to their poor bio-availability and non-specificity issues. Therefore, to outweigh the aforesaid disadvantages of the conventional strategies, there is an utmost need for the advancement of new alternative therapeutic biomaterials to treat CLI. Since past few decades, various research groups including ours have been involved in developing different pro-angiogenic nanomaterials. Among them, zinc oxide nanoflowers (ZONF), established in our laboratory, are considered as one of the potent nanoparticles to induce therapeutic angiogenesis. In our earlier studies, we have depicted that ZONF promote angiogenesis by inducing the formation of reactive oxygen species (ROS) and nitric oxide (NO) as well as activating Akt/MAPK/eNOS cell signaling pathways in the endothelial cells. Recently, we have also reported the therapeutic potential of ZONF to treat cerebral ischemia through their neuritogenic and neuroprotective properties, exploiting angio-neural cross talk. Considering the excellent pro-angiogenic properties of ZONF and importance of revascularization for the recovery of CLI, in this present study, we have comprehensively explored the therapeutic potential of ZONF in a rat hind limb ischemia model (established by ligating the femoral artery of hind limb), an animal model that mimics CLI in humans. The behavioural studies, laser Doppler perfusion imaging, histopathology, immunofluorescence as well as estimation of serum NO level depicted that the administration of ZONF could ameliorate the ischemic conditions in rats at a faster rate by promoting therapeutic angiogenesis to the ischemic sites. Altogether, the present study offers an alternative nanomedicine approach employing ZONF for the treatment of PADs.
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http://dx.doi.org/10.1088/1748-605X/abebd1DOI Listing
March 2021

Ag[Fe(CN)NO]-Fabricated Hydrophobic Cotton as a Potential Wound Healing Dressing: An Approach.

ACS Appl Mater Interfaces 2021 Mar 23;13(9):10689-10704. Epub 2021 Feb 23.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, India.

There have been reports of different types of wound dressings for various functions and purposes. Cotton being one of the most widely used wound dressing material due to its non-toxic, biodegradable, and other properties is used for fabrication as well as in the form of scaffolds for faster and effective wound closure. Our research team has already demonstrated the role of silver nitroprusside nanoparticles (SNPNPs) for wound healing and antibacterial activity. In the current study, we have developed cotton fabric impregnated with SNPNPs (SNPCFs) which remain photo inert and displayed long-term antimicrobial activity due to the surface modification with the silver nitroprusside complex. These SNPCFs were characterized by various analytical techniques (XRD, FTIR, UV spectroscopy, TGA, TEM, FESEM, EDAX, ICP-OES). The fabricated cotton dressings with nanoparticles showed an improved water contact angle (113-130°) than that of bare cotton gauze (60°) and exhibited more antibacterial property in case of both Gram-negative bacteria ( and ) and Gram-positive bacteria ( and ) even after several washings. The biocompatible nature of SNPCFs was assessed by chorioallantoic membrane assay that showed no obstruction in the formation of blood vessels. The SNPCFs exhibited better wound healing activity compared to the bare cotton and AgCFs as observed in the C57BL6/J mouse. The histopathological investigation reveals increase in re-epithelialization and deposition of connective tissue. The macrophage (M2) counts in SNPCF-treated skin tissues were supportive of more wound healing activity than mice treated with cotton fabric impregnated with chemically synthesized silver nanoparticles. Based on biodistribution analysis using ICP-OES, the data illustrated that a significant amount of silver is absorbed in the skin tissues of mice as compared to the blood and kidney. Furthermore, the absence of silver from the vital organs (heart, liver, and kidney) corroborates our hypothesis that the SNPCFs can act excellently in treating wounds when topically applied over skin. Thereafter, all these results highlight a strong possibility that SNPCFs exemplify the potential as a new antimicrobial and wound healing agent in future times.
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http://dx.doi.org/10.1021/acsami.0c19904DOI Listing
March 2021

Nanoparticle-based angiogenesis for the recovery of heavy metal-induced vascular toxicity.

Nanomedicine (Lond) 2021 02 18;16(5):351-354. Epub 2021 Feb 18.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad, Telangana 500007, India.

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http://dx.doi.org/10.2217/nnm-2021-0028DOI Listing
February 2021

Silver Prussian Blue Analogue Nanoparticles: Rationally Designed Advanced Nanomedicine for Multifunctional Biomedical Applications.

ACS Biomater Sci Eng 2020 01 30;6(1):690-704. Epub 2019 Dec 30.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana State, India.

The development of simple, cost-effective, and advanced multifunctional technology is the need of the hour to combat cancer as well as bacterial infections. There have been reports of silver nanoparticles (AgNPs), silver salts, and Prussian blue (PB) being used for medicinal purposes which are clinically approved. In this context, in the present communication, we incorporated PB and silver salts (silver nitrate) to develop silver PB analogue nanoparticles (SPBANPs), a new nanomedicine formulation as a safer and effective mode of treatment strategy (2-in-1) for both cancer and bacterial infections. Considering all fundamental issues of nanomedicine, along with understanding of the biological impact of PB, we designed a simple, fast, efficient, cheap, and eco-friendly method for the synthesis of [poly(-vinyl-2-pyrrolidone)]-stabilized silver hexacyanoferrate nanoparticles (silver PB analogue: Ag[Fe(CN)] abbreviated as SPBANPs). Various analytical tools were used to analyze and characterize the nanomaterials (SPBANPs). The SPBANPs were highly stable for several weeks in various phosphate buffers with a range of physiological pH conditions (pH = 6-8). The nanoparticles showed biocompatibility in vivo in C57BL6/J mice that encouraged us to screen the nanoparticles for various biomedical applications. The SPBANPs themselves exhibited remarkable inhibition of cancer cell proliferation (B16F10, A549, MCF-7, and SK-OV-3) in vitro. Substantial inhibition of melanoma tumor growth was observed in the C57BL6/J mouse model (aggressive murine melanoma model: B16F10) after intraperitoneal administration of the SPBANPs without any anticancer drug. Additionally, the SPBANPs exhibited excellent antibacterial activity in various Gram-negative (, , and ) and Gram-positive () bacteria. Interestingly, this nanoformulation itself works as a drug delivery vehicle, as well as an anticancer and antibacterial agent. The in vitro and in vivo results together demonstrate that this biocompatible nanoformulation (SPBANPs) without an anticancer drug or antibiotic could be explored to develop as a multifunctional therapeutic agent (2-in-1) for the treatment of cancer and bacterial infections in the near future.
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http://dx.doi.org/10.1021/acsbiomaterials.9b01693DOI Listing
January 2020

Silver Prussian blue analogue nanomedicine for future cancer therapy.

Future Oncol 2021 Jan 27;17(2):119-122. Epub 2020 Nov 27.

Department of Applied Biology, Council of Scientific & Industrial Research-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad, 500007, Telangana State, India.

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http://dx.doi.org/10.2217/fon-2020-0736DOI Listing
January 2021

Zinc oxide nanoparticles: future therapy for cerebral ischemia.

Nanomedicine (Lond) 2020 12 20;15(28):2729-2732. Epub 2020 Oct 20.

Department of Applied Biology, CSIR Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad, Telangana 500007, India.

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http://dx.doi.org/10.2217/nnm-2020-0322DOI Listing
December 2020

Vanadium pentoxide nanomaterials and their role in anti-angiogenesis for cancer treatment.

Nanomedicine (Lond) 2020 11 20;15(27):2643-2646. Epub 2020 Oct 20.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad, Telangana 500007, India.

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http://dx.doi.org/10.2217/nnm-2020-0321DOI Listing
November 2020

Fibro-porous PLLA/gelatin composite membrane doped with cerium oxide nanoparticles as bioactive scaffolds for future angiogenesis.

J Mater Chem B 2020 Sep 15. Epub 2020 Sep 15.

Biomedical Engineering and Technology Laboratory, Discipline of Mechanical Engineering, PDPM-Indian Institute of Information Technology Design and Manufacturing, Jabalpur, Dumna Airport Road, Jabalpur-482005, MP, India.

Functionalized cerium oxide nanoparticle (CeNP)-loaded fibro-porous poly-l-lactic acid (PLLA)/gelatin composite membranes were prepared via an electrospinning technology. Considering the importance of such membrane scaffolds for promoting angiogenesis in tissue engineering and drug screening, a series of PLLA/gelatin composite fiber membranes loaded with different doses of CeNPs was prepared. The prepared composite membranes demonstrated hydrophilicity, water absorption, and improved mechanical properties compared to a PLLA and PLLA/gelatin membrane. Also, cell viability assay using somatic hybrid endothelial cells (EA.hy926) proved the biocompatible nature of the scaffolds. The biocompatibility was further supported by in vivo chick embryo angiogenesis assay using fertilized eggs. Our initial results support that these membrane scaffolds could be useful for angiogenesis-related disease treatment after further investigations.
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http://dx.doi.org/10.1039/d0tb01715aDOI Listing
September 2020

Attenuation of cadmium-induced vascular toxicity by pro-angiogenic nanorods.

Mater Sci Eng C Mater Biol Appl 2020 Oct 21;115:111108. Epub 2020 May 21.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana State, India; Academy of Scientific & Innovative Research (AcSIR), Ghaziabad 201002, U.P., India. Electronic address:

Cadmium (Cd) is a common heavy metal that causes major environmental pollution with adverse effects on human health and well-being. Exposure to Cd is known to exhibit detrimental consequences on all the vital organ systems of the body, especially the vascular system. Certain approaches using anti-oxidants and chelating agents have been demonstrated previously to mitigate Cd-induced toxicity. However, these approaches are associated with their own limitations. In this context, there is a critical need for the development of alternative treatment strategies to address the conditions associated with Cd-poisoning. One such novel approach is the application of nanomedicine which is well-known to resolve several health complications by improving disease therapy. Recently, our group demonstrated the role of europium hydroxide nanorods (EHN) in promoting vascular growth using in vitro and in vivo assay systems. Therefore, in the present study, we have evaluated the effect of EHN on health of endothelial cells (EA.hy926) and fibroblasts (NIH 3T3) intoxicated by Cd. The results revealed that EHN significantly improved the viability of EA.hy926 and NIH 3T3 cells, reduced apoptotic cell population, increased nitric oxide (NO) production and promoted blood vasculature development in the chick embryo model, which were hampered due to Cd insult. Molecular studies demonstrated the reduced expression of tumor suppressor (p53) and elevated anti-apoptotic protein (Bcl-xL) levels along with enhanced NO production through endothelial nitric oxide synthase (eNOS) activation as the plausible mechanisms underlying protective role of EHN against Cd-induced vascular toxicity. Considering the above observations, we strongly believe that EHN could be a potential nanomedicine approach for overcoming Cd-induced toxicity by improving vascular health and functioning.
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http://dx.doi.org/10.1016/j.msec.2020.111108DOI Listing
October 2020

Anti-angiogenic vanadium pentoxide nanoparticles for the treatment of melanoma and their in vivo toxicity study.

Nanoscale 2020 Apr 31;12(14):7604-7621. Epub 2020 Mar 31.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad - 500007, Telangana State, India.

In recent days, vanadium complexes and nanoparticles have received sustainable attention owing to their vast applications in different fields. In the present study, we report a facile approach for the synthesis of irregular dumbbell shaped vanadium pentoxide nanoparticles (VO NPs: 30-60 nm) via the polyol-induced microwave irradiation process along with calcination. The as-synthesized nanoparticles were characterized using various physico-chemical techniques (e.g. XRD, TEM, FT-IR, DLS and XPS). The cell viability assay showed that VO NPs could efficiently inhibit the proliferation of different cancer cells (B16F10, A549, and PANC1), depicting their anti-proliferative activity. However, VO NPs did not exert significant cytotoxicity to the normal cells (CHO, HEK-293 and NRK-49F), suggesting their biocompatible nature. Interestingly, these nanoparticles inhibited the proliferation and migration of the endothelial cells (HUVECs and EA.hy926) and disrupted the blood vasculature in a chick embryo model, indicating their anti-angiogenic properties. The mechanistic study revealed that the effective internalization of VO NPs generated intracellular reactive oxygen species (ROS) which in turn up-regulated p53 protein and down-regulated survivin protein in cancer cells, leading to the apoptosis process. Furthermore, the administration of VO NPs to melanoma bearing C57BL6/J mice significantly increased their survivability as compared to the control untreated tumor bearing mice, exhibiting the therapeutic potential of the nanoparticles against melanoma. Additionally, the in vivo toxicity study demonstrated no toxic effect in mice upon sub-chronic exposure to VO NPs. Altogether, we strongly believe that VO NPs could intrinsically provide a new direction for alternative therapeutic treatment strategies for melanoma and other cancers by employing their anti-angiogenic properties in the future.
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http://dx.doi.org/10.1039/d0nr00631aDOI Listing
April 2020

Potential Therapeutic Application of Zinc Oxide Nanoflowers in the Cerebral Ischemia Rat Model through Neuritogenic and Neuroprotective Properties.

Bioconjug Chem 2020 03 25;31(3):895-906. Epub 2020 Feb 25.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, India.

Neuritogenesis, a complex process of the sprouting of neurites, plays a vital role in the structural and functional restoration of cerebral ischemia-injured neuronal tissue. Practically, there is no effective long-term treatment strategy for cerebral ischemia in clinical practice to date due to several limitations of conventional therapies, facilitating the urgency to develop new alternative therapeutic approaches. Herein, for the first time we report that pro-angiogenic nanomaterials, zinc oxide nanoflowers (ZONF), exhibit neuritogenic activity by elevating mRNA expression of different neurotrophins, following PI3K/Akt-MAPK/ERK signaling pathways. Further, ZONF administration to global cerebral ischemia-induced Fischer rats shows improved neurobehavior and enhanced synaptic plasticity of neurons via upregulation of Neurabin-2 and NT-3, revealing their neuroprotective activity. Altogether, this study offers the basis for exploitation of angio-neural cross talk of other pro-angiogenic nano/biomaterials for future advancement of alternative treatment strategies for cerebral ischemia, where neuritogenesis and neural repair are highly critical.
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http://dx.doi.org/10.1021/acs.bioconjchem.0c00030DOI Listing
March 2020

Improved delivery of doxorubicin using rationally designed PEGylated platinum nanoparticles for the treatment of melanoma.

Mater Sci Eng C Mater Biol Appl 2020 Mar 5;108:110375. Epub 2019 Nov 5.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad, 500007, Telangana State, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India. Electronic address:

Efficient delivery of chemotherapeutic drugs to tumor cells is one of the crucial issues for modern day cancer therapy. In this article, we report the synthesis of poly ethylene glycol (PEG) assisted colloidal platinum nanoparticles (PtNPs) by borohydride reduction method at room temperature. PtNPs are stable at room temperature for more than 2 years and are stable in serum and phosphate buffer (pH = 7.4) solution for one week. PtNPs show biocompatibility in different normal cell lines (in vitro) and chicken egg embryonic model (ex vivo). Further, we designed and fabricated PtNPs-based drug delivery systems (DDS: PtNPs-DOX) using doxorubicin (DOX), a FDA approved anticancer drug. Various analytical techniques were applied to characterize the nanomaterials (PtNPs) and DDS (PtNPs-DOX). This DDS exhibits inhibition of cancer cell (B16F10 and A549) proliferation, observed by different in vitro assays. PtNPs-DOX induces apoptosis in cancer cells observed by annexin-V staining method. Intraperitoneal (IP) administration of PtNPs-DOX shows substantial reduction of tumor growth in subcutaneous murine melanoma tumor model compared to control group with free drug. Up-regulation of tumor suppressor protein p53 and down regulation of SOX2 and Ki-67 proliferation markers in melanoma tumor tissues (as observed by immunofluorescence and western blot analysis) indicates probable molecular mechanism for the anticancer activity of DDS. Considering the in vitro and pre-clinical (in vivo) results in murine melanoma, it is believed that platinum nanoparticle-based drug delivery formulation could be exploited to develop an alternative therapeutic nanomedicine for cancer therapy in the near future.
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http://dx.doi.org/10.1016/j.msec.2019.110375DOI Listing
March 2020

Biosynthesized Gold Nanoparticles: In Vivo Study of Near-Infrared Fluorescence (NIR)-Based Bio-imaging and Cell Labeling Applications.

ACS Biomater Sci Eng 2019 Oct 23;5(10):5439-5452. Epub 2019 Sep 23.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana, India.

Near infrared (NIR) fluorescence imaging is a striking imaging modality for biomedical and clinical applications due to its deep tissue penetration and low phototoxicity. The major issue with NIR dyes is their non-specific distribution and requirement of tagging with biomolecules for specific tissue localization. Till now, there have been no imaging agents available that can distribute into a specific organ without the need for targeted ligands, which remains as an unmet clinical need. In the present study, we demonstrate that the plant extract (abbreviated as ZE) assisted synthesis of highly biocompatible gold nanoparticles (AuZE), leading to their non-invasive bio-imaging applications in the NIR region (red at 820 nm emission: NIR region). AuZE and ZE both exhibited green fluorescence at 350 nm excitation and red fluorescence in the NIR region (710 nm). We verified the source of this fluorescence, which originates from the fluorescent molecules present in the ZE extract. After intraperitoneal administration in C57BL6 mice, very interestingly, AuZE is distributed into the brain of C57BL6 mice without the need for any targeted ligand and exhibited bright red fluorescence in the NIR region (710 nm excitation, 820 nm emission) as evidenced by non-invasive imaging as well as ICPOES techniques. We further explored the activity of ZE and AuZE as cell labeling agents (B16F10 cells were pre-incubated with AuZE and implanted into mice, and the fluorescence was monitored), which could be applicable for graft transplantation biology. To the best of our knowledge, this is the first report that demonstrates the versatile applications of green synthesized gold nanoparticles using a ZE extract. Considering these exciting results and fruitful outcomes, the ZE and AuZE NPs would stand as an alternative imaging agent to commercially available NIR dyes and change the conventional fluorescence-based bio-imaging strategies. Therefore, the biosynthesized AuNPs open new directions for future research to explore these latest observations in the field of disease diagnosis and therapy.
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http://dx.doi.org/10.1021/acsbiomaterials.9b00721DOI Listing
October 2019

Design of DNA-intercalators based copper(II) complexes, investigation of their potential anti-cancer activity and sub-chronic toxicity.

Mater Sci Eng C Mater Biol Appl 2019 Dec 14;105:110079. Epub 2019 Aug 14.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana State, India. Electronic address:

In the present paper, we synthesized and characterized four N-donor polypyridyl copper(II) complexes (C1-C4); [Cu(mono-CN-PIP)] (C1), [Cu(tri-OMe-PIP)] (C2), [Cu(di-CF-PIP)] (C3) and [Cu(DPPZ)] (C4). The (Calf-Thymus) CT-DNA binding studies depicted that the complexes could interact with DNA via intercalative mode. All the complexes, particularly C3 and C4 attenuated the proliferation as well as migration of various cancer cells, indicating their anti-cancer and anti-metastatic activity. Additionally, chick embryo angiogenesis (CEA) assay exhibited the inhibition of vascular sprouting in presence of C3 and C4, suggesting their potential in inhibiting the blood vessel growth. Mechanistic studies revealed that the complexes induced the excessive production of cellular reactive oxygen species (ROS) leading to apoptosis through up regulation of p53 and downregulation of Bcl-xL, which might be the plausible mechanisms underlying their anti-cancer properties. To understand the feasibility of practical application of anti-cancer copper complexes C3 and C4, in vivo sub-chronic toxicity study (4 weeks) was performed in C57BL6 mice and the results exhibited almost non-toxic effects induced by these complexes in terms of haematology and serum biochemical analyses, suggesting their biocompatible nature. The current study provides the basis for future advancement of other novel biocompatible metal complexes that could be employed for the therapy of different cancers.
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http://dx.doi.org/10.1016/j.msec.2019.110079DOI Listing
December 2019

Au-CGKRK Nanoconjugates for Combating Cancer through T-Cell-Driven Therapeutic RNA Interference.

ACS Omega 2018 Aug 3;3(8):8663-8676. Epub 2018 Aug 3.

Division of Applied Biology and Analytical & Mass Chemistry Division, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana, India.

Numerous prior studies on fighting cancer have been based on using inhibitors of JAK-STAT pathway (signal transducer and activator of transcription 3 (STAT3) inhibitor in particular), a signaling pathway responsible for progression of many types of cancer cells. However, recent studies have shown that STAT3 activation leads to upregulation of program death receptor-ligand 1 (PD-L1, an immune checkpoint protein that plays a major role behind evasion of immune systems by growing tumors) expression levels in tumor cells, leading to enhanced immune suppression. This is why global efforts are being witnessed in combating cancer through use of immune checkpoint inhibitors. Herein, we report on the design, synthesis, physicochemical characterizations, and bioactivity evaluation of novel tumor- and tumor-vasculature-targeting noncytotoxic Au-CGKRK nanoconjugates (17-80 nm) for combating tumor. Using a syngeneic mouse tumor model, we show that intraperitoneal (i.p.) administration of the Au-CGKRK nanoparticles (NPs) complexed with both PD-L1siRNA (the immune checkpoint inhibitor) and STAT3siRNA (the JAK-STAT pathway inhibitor) results in significant (>70%) enhancement in overall survivability (OS) in melanoma-bearing mice ( = 5) when compared to the OS in the untreated mice group. The expression levels of CD8 and CD4 proteins in the tumor lysates of differently treated mice groups (by Western blotting) are consistent with the observed OS enhancement being a T-cell-driven process. Biodistribution study using near-infrared dye-loaded Au-CGKRK nanoconjugates revealed selective accumulation of the dye in mouse tumor. Notably, the overall survival benefits were significantly less (∼35%) when melanoma-bearing mice were treated (i.p.) with Au-CGKRK NPs complexed with only PD-L1siRNA or with STAT3siRNA alone. The presently described Au-CGKRK nanoconjugates are expected to find future use in therapeutic RNA-interference-based cancer immunotherapy.
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http://dx.doi.org/10.1021/acsomega.8b01051DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6644890PMC
August 2018

Restoration of p53 Function in Ovarian Cancer Mediated by Gold Nanoparticle-Based EGFR Targeted Gene Delivery System.

ACS Biomater Sci Eng 2019 Jul 19;5(7):3631-3644. Epub 2019 Jun 19.

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana India.

Targeted gene delivery of wild type tumor suppressor gene p53 is a promising approach to inhibit the progression of ovarian cancer. Although several gene delivery vehicles have been reported earlier, there is paucity for targeted delivery of wild type p53 to ovarian cancer using gold nanoparticles. As it is well-known that EGFR (epidermal growth factor receptor) is overexpressed in ovarian cancer, in this study we hypothesized that the FDA approved monoclonal antibody C225 (cetuximab) that targets EGFR could be used for targeted delivery of wild type p53 gene. With this impetus, we devised an approach wherein cationic gold nanoparticles (AuNPs) were employed to generate gold nanoparticle-based drug delivery system (DDS, Au-C225-p53DNA where p53DNA is pCMVp53 plasmid) that was formulated and characterized by biochemical and biophysical methods. The nanoconjugate complexed with DNA (Au-C225-p53DNA) is serum-stable and protects the bound DNA from digestion by DNase-I. Additionally, in vitro reporter gene expression assays demonstrated efficient and specific gene transfection in EGFR overexpressing SK-OV-3 cells. Further, the intraperitoneal administration of Au-C225-p53DNA in SK-OV-3 xenograft mouse model displayed significant tumor targeting and tumor regression. Altogether, these studies indicated a promising nanoparticle-based approach for targeting ovarian cancers caused by mutated p53.
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http://dx.doi.org/10.1021/acsbiomaterials.9b00006DOI Listing
July 2019

Recent advances in inorganic nanomaterials for wound-healing applications.

Biomater Sci 2019 Jul 16;7(7):2652-2674. Epub 2019 May 16.

Department of Bioengineering, Rice University, Houston, TX 77030, USA.

Chronic wounds have emerged as a major cause of mortality, especially in patients with diabetes and other pathologies. Statistics indicate that chronic wounds affect around 6.5 million patients annually, with wound care and management incurring huge economic costs. Growing incidence of chronic wounds and associated pathologies along with the limitations of current therapies have established a strong need for novel and innovative approaches to accelerate wound healing. Conventionally, chronic wounds are addressed using various FDA-approved silver-based formulations and other biomaterials. However, the toxicity associated with these conventional approaches, along with the increased frequency of chronic wound cases, makes the development of alternative therapies for effective wound healing necessary. Recently, researchers have investigated the design and development of nanoparticles, especially inorganic metal nanoparticles, as promising candidates for addressing various pathological conditions, including wound healing. Several research groups, including ours, have designed numerous metal nanoparticles (including silver, gold, zinc oxide, cerium oxide, terbium hydroxide, silica, titanium oxide, copper) and demonstrated their wound-healing properties using in vitro and in vivo models. The rise of nanotechnology-based platforms in wound healing is evidenced by the tremendous impact and number of publications observed in recent years, which has emphasized the robust potential of inorganic nanomedicine for addressing wounds. Therefore, the importance of these inorganic nanomaterial-based interventions for wound-healing applications needs to be emphasized to inform and encourage scientists and young researchers globally to engage with this expanding area of biology and medicine. In this review article, we mainly focus on highlighting the role of inorganic nanomaterials and nanomaterial-based approaches for wound healing and tissue regeneration, along with their mechanistic properties, clinical status, challenges, and future directions.
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http://dx.doi.org/10.1039/c9bm00423hDOI Listing
July 2019

Shikimoyl-ligand decorated gold nanoparticles for use in ex vivo engineered dendritic cell based DNA vaccination.

Nanoscale 2019 Apr;11(16):7931-7943

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Hyderabad 500 007, India.

Since mannose receptors (MRs) are expressed on the surfaces of dendritic cells (DCs), the most professional antigen presenting cells in our body, DNA vaccine carriers containing either covalently grafted mannosyl- or mannose-mimicking shikimoyl-ligands are being increasingly used in ex vivo DC-transfection based DNA vaccination. To this end, we have recently demonstrated that ex vivo immunization of mice with liposomes of shikimoylated cationic amphiphiles containing a 6-amino hexanoic acid spacer group in the head-group region in complexation with melanoma antigen (MART1) encoded DNA vaccine (pCMV-MART1) induces long lasting anti-melanoma immune responses (C. Voshavar, et al., J. Med. Chem., 2017, 60, 1605-1610). This finding prompted us to examine, in the present investigation, the efficacies of gold nanoparticles conjugated to the mannose-mimicking shikimoyl ligand (SL) via a 6-amino hexane thiol spacer (AuNPs-SL) for use in ex vivo DC-transfection based genetic immunization. Herein, we report on the design, synthesis, physico-chemical characterization and bioactivities of AuNPs-SL. Dynamic light scattering and transmission electron microscopy studies revealed the hydrodynamic diameters of theAuNPs-SL nanoconjugates to be within the range of 23-44 nm and their surface potentials within the range of 9-28 mV. MTT-assay showed the non-cytotoxic nature of AuNPs-SL and the findings in the electrophoretic gel retardation assays revealed strong DNA binding properties of the AuNPs-SL. Importantly, subcutaneous immunization of C57BL/6J mice with DCs ex vivo transfected with an electrostatic complex of AuNPs-SL & melanoma antigen (MART1) encoded DNA vaccine (p-CMV-MART1) induced a long lasting (100 days) anti-tumor immune response in immunized mice upon subsequent challenge with a lethal dose of melanoma. Notably, mice immunized with either autologous mbmDCs ex vivo pre-transfected with nanoplexes of shikimoylated AuNPs-SL & an irrelevant pCMV-SPORT-β-gal plasmid (without having encoded melanoma antigen) or untransfected DCs showed no lasting protection against subsequent tumor challenge. The presently described shikimoyl-decorated gold nanoparticles (AuNPs-SL) are expected to find future use in ex vivo DC-transfection based genetic immunization against cancer and other infectious diseases.
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http://dx.doi.org/10.1039/c8nr10293gDOI Listing
April 2019

In vivo targeting of DNA vaccines to dendritic cells using functionalized gold nanoparticles.

Biomater Sci 2019 Feb;7(3):773-788

Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Hyderabad 500 007, India.

The clinical success of dendritic cell (DC)-based genetic immunization remains critically dependent on the availability of effective and safe nano-carriers for targeting antigen-encoded DNA vaccines to DCs, the most potent antigen-presenting cells in the human body in vivo. Recent studies revealed the efficacies of mannose receptor-mediated in vivo DC-targeted genetic immunization by liposomal DNA vaccine carriers containing both mannose-mimicking shikimoyl and transfection enhancing guanidinyl functionalities. However, to date, the efficacies of this approach have not been examined for metal-based nanoparticle DNA vaccine carriers. Herein, we report for the first time, the design, synthesis, physico-chemical characterization and bioactivities of gold nanoparticles covalently functionalized with a thiol ligand containing both shikimoyl and guanidinyl functionalities (Au-SGSH). We show that Au-SGSH nanoparticles can deliver DNA vaccines to mouse DCs under in vivo conditions. Subcutaneous administration of near infrared (NIR) dye-labeled Au-SGSH showed significant accumulation of the NIR dye in the DCs of the nearby lymph nodes compared to that for the non-targeting NIR-labeled Au-GSH nanoconjugate containing only a covalently tethered guanidinyl group, not the shikimoyl-functionality. Under prophylactic settings, in vivo immunization (s.c.) with the Au-SGSH-pCMV-MART1 nanoplex induced a long-lasting (180 days) immune response against murine melanoma. Notably, mannose receptor-mediated in vivo DC-targeted immunization (s.c.) with the Au-SGSH-MART1 nanoplex significantly inhibited established melanoma growth and increased the overall survivability of melanoma-bearing mice under therapeutic settings. The Au-SGSH nanoparticles reported herein have potential use for in vivo DC-targeted genetic immunization against cancer and infectious diseases.
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http://dx.doi.org/10.1039/c8bm01272eDOI Listing
February 2019

Nanomedicine for Cancer Therapy Using Autophagy: An Overview.

Curr Top Med Chem 2018 ;18(30):2599-2613

Academy of Scientific and Innovative Research (AcSIR), Ghaziabad- 201002, India.

Autophagy is an intracellular biological catabolic process of mammalian cells to maintain the homeostasis. It plays a wide role in the clearance of damaged cellular organelles, misfolded or aggregated proteins like alpha-synuclein, β-amyloid peptides, Tau proteins and pathogens. Recent studies have clearly demonstrated that dysfunction in autophagy leads to the development of cancer, cardiomyopathy, chronic infection, neurodegenerative and other diseases. Therefore, modulation of autophagy has therapeutic value to cure the diseases including cancer using external stimuli. In this context, various researchers developed small molecules such as chloroquine, rapamycin, etc. for the treatment of cancer through autophagy. However, these molecules possess side effects which limit their use in the clinics. Therefore, nanomedicine approach could stand as an alternative treatment option to induce the autophagy in cancer therapy. Several investigators developed a variety of nanomaterials which themselves act as autophagy inducers or inhibitors. Considering this, the present review article will focus on the recent developments of nanomedicine in the area of autophagy that have been focused on the treatment of cancers. We also summarised the detailed mechanisms of nanoparticles mediated autophagy which could be helpful for developing new strategies to fight against cancer. Also, the present review article covers the current clinical status of nanomedicine and future challenges. Finally, we conclude with the future potential role of nanomedicine for autophagy induction in the cancer treatment.
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http://dx.doi.org/10.2174/1568026619666181224104838DOI Listing
March 2019

Ag[Fe(CN)NO] Nanoparticles Exhibit Antibacterial Activity and Wound Healing Properties.

ACS Biomater Sci Eng 2018 Sep 22;4(9):3434-3449. Epub 2018 Aug 22.

Training and Development Complex, Academy of Scientific and Innovative Research (AcSIR), CSIR Campus, CSIR Road, Taramani, Chennai 600 113, India.

Therapeutic agents harboring both wound healing and antibacterial activities have much demand in biomedical applications. Development of such candidates with clinically approved materials adds more advantages toward these applications. Recently, silver metal complex nanomaterials have been playing a major role in medical uses especially for antibacterial activity and wound healing. In this report, we designed and synthesized silver nitroprusside complex nanoparticles (abbreviated as AgNNPs) using sodium nitroprusside and silver nitrate (both are FDA approved precursors). The nanoparticles (AgNNPs) were thoroughly characterized by various physicochemical techniques such as XRD, FTIR, TGA, DLS, EDAX, Raman, ICP-OES, HRTEM, and FESEM. The cell viability assay in normal cells (EA.hy 926 cells, NIH 3T3) using MTT reagents and CEA assay (CEA: Chick embryo angiogenesis assay) in fertilized eggs demonstrate the biocompatibility of AgNNPs. These nanoparticles show effective antibacterial activity against both Gram positive and Gram negative bacteria through membrane and DNA damage. Additionally, AgNNPs accelerate the wound healing in C57BL6 mice by altering the macrophages from M1 to M2. Considering the results together, the current study may offer the development of new silver nanocomplex nanomaterials that shows synergistic effect on antibacterial activity and wound healing (2-in-1-system). To the best of our knowledge, this is the first report for the synthesis, characterization, and biomedical applications of silver nitroprusside nanoparticles.
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http://dx.doi.org/10.1021/acsbiomaterials.8b00759DOI Listing
September 2018

Engineered Nanoparticles for Effective Redox Signaling During Angiogenic and Antiangiogenic Therapy.

Antioxid Redox Signal 2019 02 24;30(5):786-809. Epub 2018 Aug 24.

1 Applied Biology Division, CSIR-Indian Institute of Chemical Technology, Hyderabad, India.

Significance: Redox signaling plays a vital role in regulating various cellular signaling pathways and disease biology. Recently, nanomedicine (application of nanotechnology in biology and medicine) has been demonstrated to regulate angiogenesis through redox signaling. A complete understanding of redox signaling pathways influenced angiogenesis/antiangiogenesis triggered by therapeutic nanoparticles is extensively reviewed in this article. Recent Advances: In recent times, nanomedicines are regarded as the Trojan horses that could be employed for successful drug delivery, gene delivery, peptide delivery, disease diagnosis, and others, conquering barriers associated with conventional theranostic approaches.

Critical Issues: Physiological angiogenesis is a tightly regulated process maintaining a balance between proangiogenic and antiangiogenic factors. The redox signaling is one of the main factors that contribute to this physiological balance. An aberrant redox signaling cascade can be caused by several exogenous and endogenous factors and leads to reduced or augmented angiogenesis that ultimately results in several disease conditions.

Future Directions: Redox signaling-based nanomedicine approach has emerged as a new platform for angiogenesis-related disease therapy, where nanoparticles promote angiogenesis via controlled reactive oxygen species (ROS) production and antiangiogenesis by triggering excessive ROS formation. Recently, investigators have identified different efficient nano-candidates, which modulate angiogenesis by controlling intracellular redox molecules. Considering the importance of angiogenesis in health care a thorough understanding of nanomedicine-regulated redox signaling would inspire researchers to design and develop more novel nanomaterials that could be used as an alternative strategy for the treatment of various diseases, where angiogenesis plays a vital role.
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http://dx.doi.org/10.1089/ars.2017.7383DOI Listing
February 2019

Multifunctional (3-in-1) cancer theranostics applications of hydroxyquinoline-appended polyfluorene nanoparticles.

Chem Sci 2017 Nov 29;8(11):7566-7575. Epub 2017 Aug 29.

Department of Chemistry , Indian Institute of Technology Guwahati , Guwahati 781039 , Assam , India . Email:

The accumulation of fluorescent hydroxyquinoline-affixed polyfluorene (PF-HQ) nanoparticles and their utility for multi-color bio-imaging and drug delivery for cancer treatment are reported. The formation of nanoparticles (PF-HQ) containing hydrophobic pockets three-dimensional growth of a polymeric backbone in a higher water fraction (THF : HO = 1 : 9) was observed. The nanoparticles showed incredible dual-state optical and fluorescence properties, which were further explored in multi-color cell imaging in both cancer and normal cells. The cell viability assay in various normal cells confirmed the biocompatible nature of PF-HQ, which was further supported by an (chick chorioallantoic membrane assay) model. This encouraged us to fabricate PF-HQ-based new drug delivery systems (DDS: PF-HQ-DOX) upon conjugation with the FDA-approved anti-cancer drug doxorubicin (DOX) by filling the hydrophobic pockets of the polymer nanoparticles. The enhanced anti-cancer activity of the DDS (PF-HQ-DOX) compared with that of free DOX was observed in mouse melanoma cancer cells (B16F10) and a subcutaneous mouse (C57BL6/J) melanoma tumor model upon administration of PF-HQ-DOX. biodistribution studies using a fluorescence quantification method demonstrated the enhanced accumulation of DOX in tumor tissues in the PF-HQ-DOX-treated group compared to that of the free drug, signifying the drug delivery efficacy of the delivery system by a passive targeting manner. Based on the above biological data ( and in the pre-clinical model), these robust and versatile fluorescent hydroxyquinoline-affixed polyfluorene (PF-HQ) nanoparticles could be effectively utilized for multifunctional biomedical applications (as they are biocompatible and can be used for bio-imaging and as a drug delivery vehicle).
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http://dx.doi.org/10.1039/c7sc03321dDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5848823PMC
November 2017

Functionalized nanoceria exhibit improved angiogenic properties.

J Mater Chem B 2017 Dec 24;5(47):9371-9383. Epub 2017 Nov 24.

Chemical Biology Division, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad - 500007, Telangana State, India.

The growth of new blood vessels from the pre-existing vasculature known as angiogenesis has a vital role in various physiological and pathological processes. In the present study, we demonstrate the pro-angiogenic properties of functional nanoconjugates of organosilane functionalized cerium oxide (CeO) nanoparticles (nanoceria). Aqueous dispersible CeO and trivalent metal (samarium) ion-doped CeO (SmCeO) nanoparticles conjugated with hydrophilic biocompatible and antifouling (6-{2-[2-(2-methoxy-ethoxy)-ethoxy]-ethoxy}-hexyl)triethoxysilane moieties were prepared. These functional nanoconjugates were prepared via an in situ synthesis and functionalization procedure using an ammonia-induced ethylene glycol-assisted precipitation method. The prepared nanoconjugates were thoroughly characterized using various physico-chemical techniques such as transmission electron microscopy (TEM), X-ray diffraction (XRD) analysis, dynamic light scattering (DLS), Fourier-transform infrared (FTIR) spectroscopy, C high-resolution solid-state nuclear magnetic resonance (NMR) spectroscopy, and X-ray photoelectron spectroscopy (XPS). The pro-angiogenic properties of the prepared nanoconjugates were evaluated by employing various angiogenesis assays (in vitro and in vivo). The results of the present study illustrate that the functional nanoconjugates of SmCeO triggered endothelial cell proliferation and induced the growth of blood vessels in a chick embryo. The enhanced expression of pro-angiogenic markers (p38 MAPK/HIF-1α) by these functional nanoconjugates might be a plausible signaling mechanism underlying their pro-angiogenic properties. Considering all the observations, we believe that (6-{2-[2-(2-methoxy-ethoxy)-ethoxy]-ethoxy}-hexyl)triethoxysilane conjugated SmCeO nanoparticles could be developed as potential candidates for the treatment of cardiovascular, ischemic and ocular diseases where angiogenesis is the principal phenomenon.
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http://dx.doi.org/10.1039/c7tb01957bDOI Listing
December 2017

Pro-angiogenic Properties of Terbium Hydroxide Nanorods: Molecular Mechanisms and Therapeutic Applications in Wound Healing.

ACS Biomater Sci Eng 2017 Dec 1;3(12):3635-3645. Epub 2017 Dec 1.

Chemical Biology Division, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana State, India.

The process of angiogenesis, involving generation of new blood vessels from the existing ones, is vital for the supply of oxygen and nutrients to various tissues of body system. Angiogenesis is directly associated with several physiological and pathological processes. It is well-established that impairment in angiogenesis process results in various fatal conditions. Recently, few research groups including ours demonstrated therapeutic angiogenesis through nanomedicine approach using metal oxide/hydroxide nanoparticles. However, there is still a thorough necessity for the development of novel, eco-friendly, pro-angiogenic nanomaterials. Hence, in the present study we demonstrate the in vitro and in vivo pro-angiogenic properties of terbium hydroxide nanorods (THNRs) synthesized using an advanced microwave irradiation method, along with the detailed molecular signaling cascade underlying THNRs induced angiogenesis. The in vivo wound healing and nonimmunogenicity of the THNRs have been validated in the mouse models. We thus strongly believe that the present study establishing the pro-angiogenic properties of THNRs will aid in the development of alternative treatment strategies for wound healing along with cardiovascular and ischemic diseases, where angiogenesis is the chief target.
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http://dx.doi.org/10.1021/acsbiomaterials.7b00457DOI Listing
December 2017

Biocompatible nickel-prussian blue@silver nanocomposites show potent antibacterial activities.

Future Sci OA 2017 Nov 6;3(4):FSO233. Epub 2017 Sep 6.

Chemical Biology Division, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana State, India.

Aim: Silver nanoparticles have long been used as potent antibacterial agents. However, toxicity concerns of silver nanoparticles have limited their successful clinical applications. Hence, development of silver-based novel biocompatible nanomaterials for antibacterial applications is a challenging task.

Materials & Methods: Accordingly, in this work, we synthesized a biocompatible silver-based nanocomposite for antibacterial applications. The nanocompostie was characterized by several analytical techniques. The nanocomposite was further tested for its cytotoxicity in cells, chicken embryo and bacteria.

Results & Conclusion: Herein, we report a simple and cost-effective method for the synthesis of nickel-prussian blue@silver nanocomposites. The nanocomposite is highly stable and shows biocompatibility observed by assay and by chicken embryonic angiogenesis assay. The nanocomposite exhibits profound antibacterial activity toward Gram-negative () and Gram-positive () bacteria. The results altogether suggest the future potential applications of nickel-prussian blue@silver nanocomposite as an antibacterial agent.
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http://dx.doi.org/10.4155/fsoa-2017-0048DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5674220PMC
November 2017

Engineered fusion protein-loaded gold nanocarriers for targeted co-delivery of doxorubicin and erbB2-siRNA in human epidermal growth factor receptor-2+ ovarian cancer.

J Mater Chem B 2017 Sep 17;5(34):7082-7098. Epub 2017 Aug 17.

Department of Chemical Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad-500007, Telangana, India.

Designed recombinant proteins comprising functional domains offer selective targeting of cancer cells for the efficient delivery of therapeutic agents. The efficacy of these carriers can be further enhanced by conjugating engineered proteins to nanoparticle surfaces. However, recombinant protein-loaded nanoparticle-based drug delivery systems are not well addressed for ovarian cancer therapy. In the present study, using a combinatorial approach, we designed and fabricated a drug delivery system by combining gold nanoparticles (AuNPs) with an engineered bi-functional recombinant fusion protein TRAF(C) (TR), loaded with an anticancer drug, namely doxorubicin (DX), and erbB2-siRNA (si), to mediate target specific delivery into SK-OV-3, a model human ovarian cancer cell line over expressing HER2 receptors (i.e. human epidermal growth factor receptor-2). The nanoparticle-based targeted drug delivery system, designated as TDDS (Au-TR-DX-si), was found to be stable and homogenous as revealed by physicochemical and biochemical studies in vitro. In addition, TDDS was functional upon evaluation in vivo. Intraperitoneal administration of TDDS at 2.5 mg kg of DX and 0.25 mg kg of erbB2 siRNA into SK-OV-3 xenograft nude mice, revealed target specific uptake and consequent gene silencing resulting in significant tumor suppression. We attribute these results to specific co-delivery of erbB2 siRNA and DX mediated by TDDS into SK-OV-3 cells via HER2 receptors. Additionally, the biodistribution of TDDS, as quantitated by ICP-OES, confirmed tumor-specific accumulation of AuNPs primarily in tumor tissues, which firmly establishes the efficacy of the nanomedicine-based combinatorial approach for the treatment of ovarian cancer in a non-toxic manner. Based on these findings, we strongly believe that the nanomedicine-based combinatorial approach can be developed as a universal strategy for treatment of HER2+ ovarian cancers.
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http://dx.doi.org/10.1039/c7tb01587aDOI Listing
September 2017

Novel tetraphenylethylene diol amphiphile with aggregation-induced emission: self-assembly, cell imaging and tagging property.

Mater Sci Eng C Mater Biol Appl 2017 Dec 12;81:580-587. Epub 2017 Aug 12.

Polymers and Functional Materials Division, CSIR-Indian Institute of Chemical Technology (CSIR-IICT), Hyderabad 500007, India; Academy of Scientific & Innovative Research (AcSIR), New Delhi. Electronic address:

In this study, we reported a novel tetraphenylethylene diol amphiphile (2) with aggregation-induced emission property which self-assembled to form unique 3D cubic superstructure. This TPE amphiphile with strong bluish green emission exhibited high biocompatibility and improved cellular uptake. Moreover, this highly fluorescent molecule was conjugated with gold nanoparticles and used as a fluorescent tagging agent. Altogether, results supported the cell-imaging and fluorescent tagging properties of tetraphenylethylene diol amphiphile.
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http://dx.doi.org/10.1016/j.msec.2017.08.051DOI Listing
December 2017

Biologically synthesized metal nanoparticles: recent advancement and future perspectives in cancer theranostics.

Future Sci OA 2017 Aug 26;3(3):FSO203. Epub 2017 May 26.

Department of Chemical Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, India.

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http://dx.doi.org/10.4155/fsoa-2017-0035DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5583654PMC
August 2017