Targeted silencing of the MCL-1 gene using multi-layered dendrimer-based nanoconstructs achieves efficient tumor regression in xenografted mice models.

The setback in the practical clinical use of RNA interference (RNAi)-based cancer treatment stems from the lack of targeted small interfering RNA (siRNA) delivery. Here, we show that luteinizing hormone-releasing hormone(LHRH) analog-tethered multi-layered polyamidoamine (PAMAM) nanoconstructs silence the anti-apoptotic MCL-1 gene in LHRH receptor overexpressing human breast (MCF-7) and prostate cancer (LNCaP) cells with 70.91 % and 74.10 % efficiency, respectively. These results were confirmed by RT-PCR. The Acridine orange/Ethidium bromide (AO/EB) dual staining revealed that the silencing of MCL-1 induced apoptosis in both the cell lines. In vivo tumor regression studies performed using MCF-7 and LNCaP xenografted severe combined immunodeficiency(SCID) mice demonstrated highly improved tumor regression in groups treated with targeted nanoconstructs complexed with MCL-1 siRNA (T + siMCL-1) compared to the other treatment groups. The quantitative RT-PCR results of tumor tissues demonstrated significant MCL-1 gene silencing, i.e., 73.76 % and 92.63 % in breast and prostate tumors, respectively, after T + siMCL-1 treatment. Reduction in MCL-1 protein expression as assessed by immunohistochemistry further confirmed these results. Furthermore, the caspase 3/7 assay demonstrated apoptosis in the MCL-1 silenced tissues. The study strongly suggests that targeted delivery of siRNAs using multi-layered dendrimer nanostructures could be an effective therapy for LHRH overexpressing cancers.

Aptamer-based Emerging Tools for Viral Biomarker Detection: A Focus on SARS-CoV-2.

Viral infections can cause fatal illnesses to humans as well as animals. Early detection of viruses is therefore crucial to provide effective treatment to patients. Recently, the Covid-19 pandemic has undoubtedly given an alarming call to develop rapid and sensitive detection platforms. The viral diagnostic tools need to be fast, affordable, and easy to operate with high sensitivity and specificity equivalent or superior to the currently used diagnostic methods. The present detection methods include direct detection of viral antigens or measuring the response of antibodies to viral infections. However, the sensitivity and quantification of the virus are still a significant challenge. Detection tools employing synthetic binding molecules like aptamers may provide several advantages over the conventional methods that use antibodies in the assay format. Aptamers are highly stable and tailorable molecules and are therefore ideal for detection and chemical sensing applications. This review article discusses various advances made in aptamer-based viral detection platforms, including electrochemical, optical, and colorimetric methods to detect viruses, specifically SARS-Cov-2. Considering the several advantages, aptamers could be game-changing in designing high-throughput biosensors for viruses and other biomedical applications in the future.

Radiation Response in the Tumour Microenvironment: Predictive Biomarkers and Future Perspectives.

Radiotherapy (RT) is a primary treatment modality for a number of cancers, offering potentially curative outcomes. Despite its success, tumour cells can become resistant to RT, leading to disease recurrence. Components of the tumour microenvironment (TME) likely play an integral role in managing RT success or failure including infiltrating immune cells, the tumour vasculature and stroma. Furthermore, genomic profiling of the TME could identify predictive biomarkers or gene signatures indicative of RT response. In this review, we will discuss proposed mechanisms of radioresistance within the TME, biomarkers that may predict RT outcomes, and future perspectives on radiation treatment in the era of personalised medicine.

Carboxymethyl fenugreek galactomannan-g-poly(N-isopropylacrylamide-co-N,N'-methylene-bis-acrylamide)-clay based pH/temperature-responsive nanocomposites as drug-carriers.

The current study dealt with the synthesis and characterization of carboxymethyl fenugreek galactomannang-g-poly(N-isopropylacrylamide-co-N,N'-methylene-bis-acrylamide)-bentonite [CFG-g-P(NIPA-co-MBA)-BEN] based nanocomposites (NCs) as erlotinib (ERL)-delivery devices for lung cancer cells to suppress excessive cell proliferation. The blank NCs exhibited outstanding biodegradability and pH/temperature-dependent swelling profiles, which were significantly influenced by their BEN contents (0-20%). The molar mass (M¯c) between the crosslinks of these NCs was declined with temperature. The composite architecture of these scaffolds was confirmed by XRD, FTIR, TGA, DSC and SEM analyses. The corresponding ERL-loaded matrices (F-1-F-3) portrayed outstanding drug encapsulation efficiency (DEE, 93-100%) with zeta potential between -8 and -16 mV and diameter between 615 and 1258 nm. These formulations demonstrated sustained ERL elution profiles (Q8h, 62-98%) with an initial burst release of drug. The drug dissolution pattern of the optimized matrices (F-3) obeyed first-order kinetic model and was driven by Fickian diffusion. The mucin adsorption behavior of F-3 was best fitted to Freudlich isotherms. The ERL-loaded formulation suppressed A549 cell proliferation and promoted apoptosis to a greater extent than the pristine drug, as detected by cellular uptake analysis, MTT cytotoxicity test and AO/EB staining assay.

Erlotinib-loaded carboxymethyl temarind gum semi-interpenetrating nanocomposites.

Erlotinib-loaded carboxymethyl temarind gum-g-poly(N-isopropylacrylamide)-montmorillonite based semi-IPN nanocomposites were synthesized and characterized for their in vitro performances for lung cancer therapy. The placebo matrices exhibited outstanding biodegradability and pH-dependent swelling profiles. The molar mass (M¯ c) between the crosslinks of these composites was declined with temperature. The solid state characterization confirmed the semi-IPN architecture of these scaffolds. The corresponding drug-loaded formulations displayed excellent drug-trapping capacity (DEE, 86-97 %) with acceptable zeta potential (-16 to -13 mV) and diameter (967-646 nm). These formulations conferred sustained drug elution profiles (Q8h, 77-99 %) with an initial burst release. The drug release profile of the optimized formulation (F-3) was best fitted in the first order kinetic model with Fickian diffusion driven mechanism. The mucin adsorption to F-3 followed Langmuir isotherms. The results of MTT assay, AO/EB staining and confocal analyses revealed that the ERL-loaded formulation suppressed A549 cell proliferation and induced apoptosis more effectively than pristine drug.

miRNA transfection via poly(amidoamine)-based delivery vector prevents hypoxia/reperfusion-induced cardiomyocyte apoptosis.

Aim: Myocardial infarction is a tissue injury that leads to apoptosis of cardiomyocytes. This can be prevented by using miRNAs, but its delivery to cardiomyocytes is a major hurdle. We aimed to deliver miRNAs using poly(amidoamine)-histidine (PAMAM-His) nanocarriers to prevent apoptosis. Materials & methods: The PAMAM-His nanoparticles were synthesized and assessed for their transfection efficiency of miRNAs to prevent apoptosis in hypoxia/reperfusion-induced H9c2 as well as primary cultured cardiomyocytes. Results & conclusion: miRNAs-nanoparticle complexes exerted a significant antiapoptotic effect on the H9c2 and primary rat ventricular cardiomyocytes. Enhanced expression of antiapoptotic genes and decreased expression of proapoptotic genes were observed. PAMAM-His nanoparticles effectively delivered miRNAs to the cardiomyocytes and prevented the hypoxia/reperfusion-induced apoptosis critical in myocardial infarctions.

Smart triblock dendritic unimolecular micelles as pioneering nanomaterials: Advancement pertaining to architecture and biomedical applications.

Recent interest in triblock dendritic unimolecular micelles has opened a new spectrum for its ubiquitous application in biomedical sciences specially drug delivery. Unimolecular dendritic micelles have brought significant attention due to their high encapsulation efficiency, high-functionality, and site specific confinement capabilities. During the last decade, the number of publications in this field has increased drastically, reinforced by the fact that several clinical trials are underway using micelles for drug delivery. This review unveils the most recent advancement on the synthesis and applications of cutting-edge unimolecular micelles using dendritic and star-shaped molecules emphasizing on triblock copolymers. The major biomedical applications covered in this manuscript are drug/gene/bio-macromolecules delivery to the site of interest, as contrast agents in bio-imaging, and cancer targeted theranostics using stimuli-responsive mechanism.

Decapeptide functionalized targeted mesoporous silica nanoparticles with doxorubicin exhibit enhanced apoptotic effect in breast and prostate cancer cells.

BACKGROUND: Considering the increase in cancer cases and number of deaths per year worldwide, development of potential therapeutics is imperative. Mesoporous silica nanoparticles (MSNPs) are among the potential nanocarriers having unique properties for drug delivery. Doxorubicin (DOX), being the most commonly used drug, can be efficiently delivered to gonadotropin-releasing hormone (GnRH)-overexpressing cancer cells using functionalized MSNPs. AIM: We report the development of decapeptide-conjugated MSNPs loaded with DOX for the targeted drug delivery in breast and prostate cancer cells. MATERIALS AND METHODS: MSNPs were synthesized and subsequently functionalized with an analog of GnRH by using a heterobifunctional polyethylene glycol as a linker. These targeted MSNPs were then characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and Raman spectroscopy. An anticancer drug DOX was loaded and then characterized for drug loading. DOX-loaded nanocarriers were then studied for their cellular uptake using confocal microscopy. The cytotoxicity of DOX-loaded targeted MSNPs and DOX-loaded bare MSNPs was studied by performing MTT assay on MCF-7 (breast cancer) and LNCaP (prostate cancer) cells. Further, acridine orange/ethidium bromide staining, as well as flow cytometry, was performed to confirm the apoptotic mode of cancer cell death. RESULTS: MSNPs were conjugated with polyethylene glycol as well as an agonist of GnRH and subsequently loaded with DOX. These targeted and bare MSNPs showed excellent porous structure and loading of DOX. Further, higher uptake of DOX-loaded targeted MSNPs was observed as compared to DOX-loaded bare MSNPs in GnRH-overexpressing breast (MCF-7) and prostate (LNCaP) cancer cells. The targeted MSNPs also showed significantly higher (P<0.001) cytotoxicity than DOX-loaded bare MSNPs at different time points. After 48 hours of treatment, the IC50 value for DOX-loaded targeted MSNPs was found to be 0.44 and 0.43 µM in MCF-7 and LNCaP cells, respectively. Acridine orange/ethidium bromide staining and flow cytometry analysis further confirmed the pathway of cell death through apoptosis. CONCLUSION: This study suggests GnRH analog-conjugated targeted MSNPs can be the suitable and promising approach for targeted drug delivery in all hormone-dependent cancer cells.

Mesoporous silica nanoparticles as cutting-edge theranostics: Advancement from merely a carrier to tailor-made smart delivery platform.

Large surface area, uniform and tunable pore size, high pore volume and low mass density- such attractive features of Mesoporous silica nanoparticles (MSNPs) have compelled researchers to explore the biomedical potential of this nano-material. Recently gained interest in MSNPs have been due to their tremendous potential in cancer therapy and imaging. Last several years have witnessed a rapid development in engineering functionalized MSNPs with various types of functional groups integrated into the system for imaging and therapeutic applications. Although their potential for drug delivery application has been studied since the year 2000, still a major challenge is to improve drug loading capacity and in vivo targeting with minimal side-effects to major organs. In this review article, the recent development of MSNPs as a therapeutic and diagnostic platform has been detailed out with emphasis on drug and bio-macromolecule delivery/co-delivery, bio-imaging and detoxification.

Triptorelin Tethered Multifunctional PAMAM-Histidine-PEG Nanoconstructs Enable Specific Targeting and Efficient Gene Silencing in LHRH Overexpressing Cancer Cells.

Cancer treatment using siRNA based therapies pose various limitations such as off-target effects and degradation due to lack of specific delivery in desired cells. The aim of the present study was to develop multifunctional targeted nanoconstructs, which can efficiently and precisely deliver siRNA and silence the desired gene of interest in various LHRH overexpressing cancer cells. Herein, we report the development of triblock, PAMAM-histidine-PEG dendritic nanoconstructs functionalized with triptorelin (an LHRH analog) for targeted siRNA delivery to LHRH overexpressing breast (MCF-7) and prostate (LNCaP) cancer cells. The nanoconstructs were characterized using 1H NMR and DLS and displayed a very low cationic charge to avoid off-target interactions. The developed nanoconstructs showed negligible cytotoxicity and hemolytic activity with efficient siRNA loading, excellent serum stability, and strongly protected siRNA from degradation. Further, confocal microscopy results confirmed extremely significant (p < 0.001) higher cellular uptake of cy5.5 conjugated targeted nanoparticles (NPs) in both cancer cell lines than nontargeted NPs. Also, targeted NPs specifically delivered cy3-tagged siRNA to MCF-7 cells. Co-localization studies in MCF-7 and LNCaP cells further established that targeted NPs traveled through the endolysosomal pathway and escaped endosomes within 6 h of incubation. Gene silencing studies in luciferase expressing MCF-7 and LNCaP cell lines demonstrated that the targeted NPs exhibited extremely significant (p < 0.001) silencing of luciferase gene. Additionally, receptor blockade studies further confirmed the specificity of targeted NPs and suggested that targeted NPs entered cancer cells via LHRH receptor mediated endocytosis, which was evident through insignificant gene silencing in receptor blocked cells. Thus, the results indicated that PAMAM-histidine-PEG-triptorelin could be a promising approach for siRNA delivery, gene silencing, and tumor therapy in all LHRH overexpressing cancer cells.

Folate/N-acetyl glucosamine conjugated mesoporous silica nanoparticles for targeting breast cancer cells: A comparative study.

Folate receptors (FR) have been well recognized as a marker to target nano-sized carriers for cancer diagnosis and therapy. In contrast, influx transport systems (e.g. GLUT transporters) that transport essential amino acids and nutrients to cancer cells have not been exploited much for targeted delivery. In this study, folic acid- or n-acetyl glucosamine- functionalized mesoporous silica nanoparticles loaded with doxorubicin (DOX-FA-MSNPs or DOX-NAG-MSNPs) were prepared, characterized and compared for targeting along with cytotoxicity towards MCF-7 and MDA-MB-231 human breast cancer cells. Cellular uptake of FITC tagged FA-MSNPs and NAG-MSNPs were evaluated by confocal microscopy and flow cytometry in above-mentioned cancer cell lines. The result suggested higher cellular uptake of NAG-MSNPs than FA-MSNPs for both the cell lines. Cytotoxicity of free DOX, DOX-MSNPs, DOX-FA-MSNPs and DOX-NAG-MSNPs were evaluated on both the breast cancer cell lines. Cytotoxicity results showed that DOX-loaded NAG-MSNPs exerted significant higher cytotoxicity effect on both the cell lines than DOX-FA-MSNPs. Moreover, both the targeted formulations were more effective than free DOX. Our results suggested that GLUT transporters can be effectively utilized for nanoparticles internalization in breast cancer cells.