Site-specific delivery of chemotherapeutics specifically to neoplastic hepatocytes without affecting normal hepatocytes should be a focus for potential therapeutic management of hepatocellular ...carcinoma (HCC). The aptamer TLS 9a with phosphorothioate backbone modifications (L5) has not been explored so far for preferential delivery of therapeutics in neoplastic hepatocytes to induce apoptosis. Thus, the objective of the present investigation was to compare the therapeutic potential of L5-functionalized drug nanocarrier (PTX-NPL5) with those of the other experimental drug nanocarriers functionalized by previously reported HCC cell-targeting aptamers and non-aptamer ligands, such as galactosamine and apotransferrin. A myriad of well-defined investigations such as cell cycle analysis, TUNEL (terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick end labeling) assay, and studies related to apoptosis, histopathology, and immunoblotting substantiated that PTX-NPL5 had the highest potency among the different ligand-attached experimental formulations in inducing selective apoptosis in neoplastic hepatocytes via a mitochondrial-dependent apoptotic pathway. PTX-NPL5 did not produce any notable toxic effects in healthy hepatocytes, thus unveiling a new and a safer option in targeted therapy for HCC. Molecular modeling study identified two cell-surface biomarker proteins (tumor-associated glycoprotein 72 TAG-72 and heat shock protein 70 HSP70) responsible for ligand-receptor interaction of L5 and preferential internalization of PTX-NPL5 via clathrin-mediated endocytosis in neoplastic hepatocytes. The potential of PTX-NPL5 has provided enough impetus for its rapid translation from the pre-clinical to clinical domain to establish itself as a targeted therapeutic to significantly prolong survival in HCC.
Display omitted
Hepatocellular carcinoma (HCC) is still a leading cause of cancer-related death worldwide. But its chemotherapeutic options are far from expectation. We here compared H-ras targeted genetic therapy ...to a commercial docetaxel formulation (DXT) in inhibiting HCC in rats.
After the physicochemical characterization of phosphorothioate-antisense oligomer (PS-ASO) against H-ras mutated gene, the PS-ASO-mediated in vitro hemolysis, in vivo hepatic uptake, its pharmacokinetic profile, tissue distribution in some highly perfused organs, its effect in normal rats, antineoplastic efficacy in carcinogen-induced HCC in rats were evaluated and compared against DXT treatment. Mutated H-ras expression by in situ hybridization, hep-par-I, CK-7, CD-15, p53 expression patterns by immunohistochemical methods, scanning electron microscopic evaluation of hepatic architecture, various hepatic marker enzyme levels and caspase-3/9 apoptotic enzyme activities were also carried out in the experimental rats.
PS-ASO showed low in vitro hemolysis (<3 %), and had a sustained PS-ASO blood residence time in vivo compared to DTX, with a time-dependent hepatic uptake. It showed no toxic manifestations in normal rats. PS-ASO distribution was although initially less in the lung than liver and kidney, but at 8 h it accumulated more in lung than kidney. Antineoplastic potential of PS-ASO (treated for 6 weeks) excelled in inhibiting chemically induced tumorigenesis compared to DTX in rats, by inhibiting H-ras gene expression, some immonohistochemical modulations, and inducing caspase-3/9–mediated apoptosis. It prevented HCC-mediated lung metastatic tumor in the experimental rats.
PS-ASO genetic therapy showed potential to inhibit HCC far more effectively than DXT in rats.
Display omitted
•Antisense oligomer (PS-ASO) increased blood half-life, and time-dependent hepatic uptake and lung accumulation in rats.•It did not show any toxicity in normal rats.•PS-ASO therapy showed superior therapeutic potential to docetaxel in inhibiting hepatocellular carcinoma in rats.•It showed antineoplastic potential, inhibiting H-ras gene expression and inducing caspase-3/9.•PS-ASO also inhibited metastatic tumor development potential of HCC in lungs.
Ormeloxifene (OLF) is an estrogen receptor modulator used in breast cancer treatment with highly favorable pharmacodynamics and pharmacokinetic properties. Folic acid (FA) receptor overexpresses in ...breast cancer cells. Hence, the study was intended to evaluate the therapeutic potential of OLF-encapsulated FA-peptide-conjugated nanoparticles (FA-Pep-OLF-NP) for breast cancer treatment.
We prepared folic acid tripeptide conjugated OLF-encapsulated PLGA nanoparticles. The surface conjugation of folic acid-tripeptide was assessed by FTIR and XPS analysis. Selectivity, cytotoxicity, apoptosis, mitochondrial depolarization, reactive oxygen species, and cell cycle analysis of the experimental nanoparticles were tested on human breast cancer cells, MDA-MB-231 and MCF-7. The therapeutic efficacy of FA-Pep-OLF-NP and unconjugated nanoparticle OLF-NP was compared in the MDA-MB-231-xenograft tumor animal model.
OLF-NP and FA-Pep-OLF-NP exhibited 4.8 % and 4.5 % w/w of drug loading, respectively. The particles were spherical with a smooth surface and uniform size distribution. FA-Pep-OLF-NP revealed higher cytotoxicity, mitochondrial depolarization, and apoptosis potential in MDA-MB-231 cells compared to MCF-7 cells. Tumor uptake and tumor volume reduction by FA-Pep-OLF-NP and OLF-NP were established in MDA-MB-231 tumor-bearing nude mice.
FA-Pep-OLF-NP showed preferential delivery of OLF to the breast cancer cells and in breast tumors compared to OLF-NP and showed more drug accumulation in breast cancer xenograft models. FA-Pep-OLF-NP delayed the progress of folate receptor overexpressed breast cancer.
Ormeloxifene (OLF) nanoparticles conjugated with folic acid-tripeptide exhibited significant in apoptosis and reduced MDA-MB-231 xenograft tumor. Abbreviation: (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, Hexafluorophosphate Benzotriazole Tetramethyl Uronium) (HBTU), Hydroxybenzotriazole (HOBt), N,N-Diisopropylethylamine (DIPEA). Display omitted
•Folic acid peptide surface functionalized nanocarrier (FA-Pep-OLF-NP) exhibited apoptosis in MDA-MB-231 and MCF-7 cells.•FA-Pep-OLF-NP were non-toxic to normal cells, and effective in the treatment of breast cancer.•99m-Tc-FA-Pep-OLF-NP accumulated more in the MDA-MB-231 bearing tumor in mice.
Aspasomes of methotrexate with antioxidant, ascorbyl palmitate, were developed and optimized using factorial design by varying parameters such as lipid molar ratio, drug to lipid molar ratio, and ...type of hydration buffer for transdermal delivery for disease modifying activity in rheumatoid arthritis (RA). Aspasomes were characterized by drug-excipients interaction, particle size analysis, determination of zeta potential, entrapment efficiency, and surface properties. The best formulation was loaded into hydrogel for evaluation of
in vitro
drug release and tested
in vivo
against adjuvant induced arthritis model in wistar rats, by assessing various physiological, biochemical, hematological, and histopathological parameters. Optimized aspasome formulation exhibited smooth surface with particle size 386.8 nm, high drug loading (19.41%), negative surface potential, and controlled drug release
in vitro
over 24 h with a steady permeation rate. Transdermal application of methotrexate-loaded aspasome hydrogel for 12 days reduced rat paw diameter (21.25%), SGOT (40.43%), SGPT (54.75%), TNFα (33.99%), IL β (34.79%), cartilage damage (84.41%), inflammation (82.37%), panus formation (84.38%), and bone resorption (80.52%) as compared to arthritic control rats. Free methotrexate-treated group showed intermediate effects. However, drug-free aspasome treatment did not show any effect. The experimental results indicate a positive outcome in development of drug-loaded therapeutically active carrier system which presents a non-invasive controlled release transdermal formulation with good drug loading, drug permeation rate, and having better disease modifications against RA than the free drug, thereby providing a more attractive therapeutic strategy for rheumatoid disease management.
Aptamers offer a significant promise to target various cancers including hepatocellular carcinoma (HCC), for their high affinity and ability to reach the target site(s), non-immunogenicity, and low ...cost. The targeting ability to neoplastic hepatocytes by the aptamer, TLS 9a with phosphorothioate backbone modification (designated as L5), has not been explored yet. Hence, we investigated the comparative potential of L5 with some other previously reported liver cancer cell-specific aptamers, conjugated on the surface of drug-nanocarriers. Various
in vitro
studies such as cytotoxicity,
in vitro
cellular uptake, cell cycle analysis, and investigations related to apoptosis were performed.
In vivo
studies carried out here include macroscopic and microscopic hepatic alterations in chemically induced hepatocarcinogenesis in rats, upon experimental treatments. The outcome of the investigations revealed that L5-functionalized drug-nanocarrier (PTX-NPL5) had the highest apoptotic potential compared with the other aptamer-conjugated experimental formulations. Further, its maximum internalization by neoplastic hepatocytes and minimum internalization by normal hepatocytes indicate that it had the potential to preferentially target the neoplastic hepatocytes. Data of
in vivo
studies revealed that PTX-NPL5 reduced tumor incidences and tumor progress. Superior potency of PTX-NPL5 may be due to the maximum affinity of L5 towards neoplastic hepatocytes resulting in maximum permeation of drug-nanocarrier in them. An effective site-specific targeting of neoplastic hepatocytes can be achieved by L5 for preferential delivery of therapeutics. Further, investigations are needed to identify the target protein(s) on neoplastic hepatocytes responsible for ligand-receptor interaction of L5.
This research aims to compare the therapeutic potential of target-specific phosphorothioate backbone-modified aptamer L5 (TLS9a)-functionalized paclitaxel (PTX)-loaded nanocarrier (PTX-NPL5) that we ...formulated with that of non-targeted commercial formulation, protein albumin-bound nanoparticles of PTX, Abraxane® (CF) against hepatocellular carcinoma (HCC) through a myriad of preclinical investigations.
A variety of in vitro and in vivo assays have been executed to compare the therapeutic effects of the formulations under investigation, including the investigation of the degree of apoptosis induction and its mechanism, cell cycle analysis, the level of ROS production, and redox status, the morphological and histological characteristics of malignant livers, and in vivo imaging. The formulations were also compared concerning pharmacokinetic behaviors. Finally, in silico molecular docking has been performed to predict the possible interactions between aptamer and target(s).
PTX-NPL5 exhibited therapeutic superiority over CF in terms of inducing apoptosis, cell cycle arrest, endorsing oxidative stress to neoplastic cells, and reducing hepatic cancerous lesions. Unlike CF, PTX-NPL5 did not exhibit any significant toxicity in healthy hepatocytes, proving enough impetus regarding the distinctive superiority of PTX-NPL5 over CF. The pharmacokinetic analysis further supported superior penetration and retention of PTX-NPL5 in neoplastic hepatocytes compared to CF. A molecular modeling study proposed possible interaction between aptamer L5 and heat shock protein 70 (HSP70).
The target-specificity of PTX-NPL5 towards neoplastic hepatocytes, probably achieved through HSP70 recognition, enhanced its therapeutic efficacy over CF, which may facilitate its real clinical deployment against HCC in the near future.
Display omitted
•Aptamer L5-functionalized drug-nanocarrier (PTX-NPL5) has superior therapeutic efficacy over commercial formulation in HCC.•PTX-NPL5 was found to have no notable toxicity in healthy hepatocytes.•Molecular modeling identified aptamer L5 efficiently interacted with HSP70 resulting PTX-NPL5 internalization in HCC tumor.
To evaluate the chemotherapeutic efficacy of J591 fabricated poly(d,l)-lactic-co-glycolic acid (PLGA) nanoparticles containing paclitaxel (Ab-PTX-NP) in vitro in PSMA (prostate specific membrane ...antigen) expressing prostate cancer cells, increase the solubility, bioavailability, circulation time, and limit systemic toxicity to achieve the maximum curative effect accompanied by controlled dosing, we formulated Ab-PTX-NP. Physicochemical characterizations such as Field emission scanning electron microscopy, Transmission electron microscopy, and Atomic force microscopy revealed that the particles were smooth-surfaced, with homogeneous distribution of drug within the particles and size were in the nano range. The encapsulation efficiency of Ab-PTX-NP was found to be 70.85%. This study acknowledges the effectiveness of Ab-PTX-NP in vitro, which displays elevated cellular cytotoxicity and internalization, maximum apoptosis (74.1%) in PSMA-abundant LNCaP cells, in comparison to PSMA negative PC3 cells. Pharmacokinetic data revealed the bioavailability of paclitaxel upon i.v. administration in the systemic circulation of male Balb/c mice. Herein, J591 was maneuvered in a neoteric way to carry the prepared chemotherapeutic nanoparticles directly to the affected prostate cancer cells.
Display omitted
•Anti-PSMA J591 paclitaxel PLGA nanoparticle was developed.•It targeted PSMA positive prostate cancer cells.•It induced apoptosis in them.
The prediction of precipitated graphite nodules size and distribution in a large industrial casting is critical to understand the mechanical behavior of cast iron components used in heavy vehicles. ...An accurate prediction of the graphite nodules requires a validated and integrated macro-micro modeling framework, which forms the motivation behind the present study. Classical theories in the literature (Lesoult
et al
. in Acta Mater 46:983–995, 1998) proposed two stages of graphite growth: in (i) liquid stage, after encapsulation by the austenite grain, and in (ii) solid stage, surrounded by only austenite phase. In this work, a new stage of graphite growth was proposed, where a graphite nodule was in direct contact with the liquid metal, existing in the presence of an austenite grain separated from the nodule. The resulting three-stage graphite growth in a microscopic control volume was formulated using a volume-averaged micro-model. This was made to evolve with the help of a macroscopic temperature field obtained from finite-element-based numerical simulation and thus creating a comprehensive modeling framework. Further, for the first time, a diffusion-based deforming-grid micro-model was developed to obtain the exact nature of a single graphite nodule growth based on the position of individual phases in the microscopic control volume. The model predictions were validated with experimental results from the step-casting experiments in the present study, as well as with the observations of single nodule growth from
in situ
synchrotron X-ray tomography (Bjerre
et al
. in Model Simul Mater Sci Eng 26:085012, 2018; Azeem
et al
. in Acta Mater 155:393–401, 2018). The proposed models captured, faithfully, the experimental patterns of graphite growth evolution, number density of the nodules, and the size distribution as a function of cooling rate. This integrated multi-scale modeling approach is envisaged to be effective for determining exact graphite growth behavior of a single nodule and volume-averaged graphite growth in a large casting.
PDF
