Background:Iron toxicity is a major contributor to adverse pregnancy outcomes in women with transfusion-dependent thalassemia. Currently used iron chelators are not recommended during pregnancy, as they can cross the placenta causing potential risk to the fetus. However, ceasing medication may adversely affect the mother's health in both the short- and long-term. Objective:We previously demonstrated that melanin nanoparticles can effectively chelate iron, and this has been confirmed by others in iron-overloaded mice. This study aims to assess whether these nanoparticles cross the placenta and evaluate their biocompatibility and haemocompatibility. Study Design:A library of 50 nm, 200 nm, and 500 nm melanin nanoparticles were synthesized and coated with Polyethylene Glycol (PEG) to improve their stability. The particles were tested for chelating iron efficacy in and biocompatibility. An in vitro BeWo (choriocarcinoma) cell model and ex vivo human placental perfusion system were used to assess nanoparticle transplacental passage. Results:Melanin nanoparticles of all sizes were able to chelate iron with a maximum adsorption of 14 mm iron/g of material; significantly higher than Desferrioxamine (DFO) of the same concentration. It was also determined that PEGylated melanin nanoparticles with appropriate size (cut off 200 nm) could be restricted from passing across the placental barrier in an in vitro model using a human choriocarcinoma cell line and in an ex vivo human placental perfusion model. The particles did not cause red cell haemolysis or blood clotting at concentrations up to 1 mM. Conclusion:It was demonstrated herein that transport of MNPs across the placental barrier is highly dependent on particle size (cut off size of 200 nm PEGylated MNPs). Findings suggest the possibility of providing a safe method of iron chelation during pregnancy. Future work using in vivo models will be applied to study systemic particle interactions.

Radio waves are highly penetrating, non-ionizing, and cause minimal damage to surrounding tissues. Radio wave control of drug release has been achieved using a novel thermoresponsive copolymer bound to a superparamagnetic iron oxide nanoparticle (SPION) core. A NIPAM-acrylamide-methacrolein copolymer underwent a coil-to-globular structure phase change upon reaching a critical temperature above the human body temperature but below hyperthermic temperatures. The copolymer was covalently bound to SPIONs which increase in temperature upon exposure to radio waves. This effect could be controlled by varying input energies and frequencies. For controlled drug release, proteins were bound via aldehyde groups on the copolymer and amine groups on the protein. The radio wave-induced heating of the complex thereby released the drug-bearing proteins. The fine-tuning of the radio wave exposure allowed multiple cycles of protein-drug release. The fluorescent tagging of the complex by FITC was also achieved in situ, allowing the tagging of the complex. The localization of the complex could also be achieved in vitro under a permanent magnetic field.

Gold nanoparticles (GNPs) possess various interesting plasmonic properties that can provide a variety of diagnostic and therapeutic functionalities for biomedical applications. Compared to other inorganic metal nanoparticles (NPs), GNPs are less toxic and more biocompatible. However, the in vivo toxicity of gold nanoparticles on humans can be significant due to the size effect. This work aims to study the effect of multiple doses of small-size (≈20 nm) GNPs on the vital organs of Wistar rats. The study includes the oxidative stress in vital organs (liver, brain, and kidney) caused by GNPs and histopathology analysis. The rats were given a single caudal injection of NPs dispersed in PBS at 25, 50, 100, and 250 mg/kg of body weight. After sacrifice, both plasma and organs were collected for the determination of oxidant/antioxidant markers and histological studies. Our data show the high sensitivity of oxidative stress parameters to the GNPs in the brain, liver, and kidneys. However, the response to this stress is different between the organs and depends upon the antioxidant defense, where GSH levels control the MDA and PCO levels. Histological alterations are mild at 25, 50, and 100 mg/kg but significant at higher concentrations of 250 mg/kg. Therefore, histological impairments are shown to be dependent on the dose of GNPs. The results contribute to the understanding of oxidative stress and cellular interaction induced by nanoparticles.

NELL1 (Neural epidermal growth factor-like (EGFL)-like protein) is an important biomarker associated with tissue and bone development and regeneration. NELL1 upregulation has been linked with metastasis and negative prognosis in rhabdomyosarcoma (RMS). Furthermore, multiple recent studies have also shown the importance of NELL1 in inflammatory bowel disease and membranous nephropathy, amongst other diseases. In this study, several anti-NELL1 DNA aptamers were selected from a randomized ssDNA pool using a fluorescence-guided method and evaluated for their binding affinity and selectivity. Several other methods such as a metabolic assay and confocal microscopy were also applied for the evaluation of the selected aptamers. The top three candidates were evaluated further, and AptNCan3 was shown to have a binding affinity up to 959.2 nM. Selectivity was examined in the RH30 RMS cells that overexpressed NELL1. Both AptNCan2 and AptNCan3 could significantly suppress metabolic activity in RMS cells. AptNCan3 was found to locate on the cell membrane and also on intracellular vesicles, which matched the location of NELL1 shown by antibodies in previous research. These results indicate that the selected anti-NELL1 aptamer showed strong and highly specific binding to NELL1 and therefore has potential to be used for in vitro or in vivo studies and treatments.

Metastatic alveolar rhabdomyosarcoma (aRMS) is an aggressive paediatric cancer with a poor prognosis. Downregulation of critical tumour genes using targeted siRNA remains an obstacle, but association with nanoparticles could help to deliver, protect, target, and enhance penetration. siRNA towards two genes was investigated: (i) Human αB-crystallin (CRYAB) and Heat Shock Protein Family B (Small) Member 2 (HSPB2), and (ii) Keratin 17 (KRT17). A mesoporous silica based nanosystem was linked to siRNA via disulfide bonds and loaded with IR820 dye. Transfection efficiency and signalling was evaluated, and the metabolic effects and cell proliferation were monitored in 2D culture and 3D spheroid models. The bound siRNA was protected from degradation with RNase I for at least 24 h. The delivered siRNA showed significant suppression of viability; 53.21 ± 23.40% for CRYAB and HSPB2 siRNA, and 88.06 ± 17.28% for KRT17 siRNA. After 72 h this increased to >50% cell apoptosis and necrosis. Intracellular total glutathione (GSH) levels were also compared with fibroblasts, and the RMS cell lines showed a several-fold increase. IR820 cellular uptake rate and penetration depth was significantly improved by nanoparticle delivery. Targetted siRNA delivery may pave the way for less invasive and more effective treatments of aRMS.

The critical role of dysregulated epigenetic pathways in cancer genesis, development, and therapy has typically been established as a result of scientific and technical innovations in next generation sequencing. RNA interference, histone modification, DNA methylation and chromatin remodelling are epigenetic processes that control gene expression without causing mutations in the DNA. Although epigenetic abnormalities are thought to be a symptom of cell tumorigenesis and malignant events that impact tumor growth and drug resistance, physicians believe that related processes might be a key therapeutic target for cancer treatment and prevention due to the reversible nature of these processes. A plethora of novel strategies for addressing epigenetics in cancer therapy for immuno-oncological complications are currently available - ranging from basic treatment to epigenetic editing. - and they will be the subject of this comprehensive review. In this review, we cover most of the advancements made in the field of targeting epigenetics with special emphasis on microbiology, plasma science, biophysics, pharmacology, molecular biology, phytochemistry, and nanoscience.

Iron toxicity is a major contributor to adverse pregnancy outcomes in women with transfusion-dependent thalassemia. Currently used chelators are not recommended during pregnancy, as they can cross the placenta causing potential risk to the foetus. Ceasing medication may adversely affect the mother's health even long after delivery. Previous experiments in iron-overloaded mice have shown that melanin nanoparticles (MNPs) can effectively chelate iron. The aim of this study was to determine whether a nanoparticle can be restricted from movement across the placenta knowing that interaction between nanoparticles with cells and tissues is determined by factors that can be readily manipulated at the synthesis stage. A library of 50, 200 and 500 nm MNPs were synthesized and coated with Polyethylene Glycol to improve their stability in circulation. Particles were then tested for their efficacy in chelating iron and for cellular and blood toxicity (n=6). An in-vitro model using Bewo cell lines (n=3) followed by an ex-vivo human placenta perfusion (n=4) was used to determine if any of the particles can pass through the placental barrier alongside Antipyrine and FITC-dextran as positive and negative controls, respectively. MNPs of all sizes were able to chelate iron with maximum absorption of 14 mM of iron/g of material; significantly higher than DFO of the same concentration (figure 1). We also found that MNPs with appropriate size (cut off 200nm) can be restricted from moving across the placental barrier in an in vitro model using human choriocarcinoma cell lines (n=3) (figure 2) and the ex-vivo human placental perfusion (n=4). MNPs showed neither haemotoxicity, nor significant cell toxicity compared to DFO when incubated with BeWo cells (over 88% viability vs. 60% for DFO). We have shown here that the transport of MNPs across the placental barrier is highly dependent on particle size. Further work on particle uptake, and in-vivo models will be applied to provide a therapeutic option for iron-overload in β-thalassemia that is safe to use during pregnancy.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Confining chemotherapy to tumour sites by means of active targeting nanoparticles (NPs) may increase the treatment effectuality while reducing potential side effects. Cubosomes are one of the next-generation drug delivery nanocarriers by virtue of their biocompatibility and bioadhesion, sizeable payload encapsulation and high thermostability. Herein, an active tumour targeting system towards rhabdomyosarcoma (RMS) cells was evaluated. Cubosomes were loaded with helenalin (a secondary metabolite from Arnica plants), which we have previously shown to induce apoptosis in RMS cells. The functionalization of the cubosomes was accomplished to enable binding to membrane receptors and translocation under a magnetic field. RMS cells overexpress CD44 and CD221 on their membrane surface and, therefore, hyaluronic acid (HA, a ligand for CD44) and antibodies (Abs) against CD221 were coupled to cubosomes via electrostatic attraction and the thiol-Michael reaction, respectively. Magnetization of the cubic phase NPs was achieved by embedding superparamagnetic iron oxide NPs (SPIONPs) into the cubic matrix. Single-function and multi-function cubosomes had Im3m cubic phase structures with well-organized lattice patterns. Conjugation with 2% HA or anti-CD221 half Abs and/or 1% SPIONPs showed significantly higher uptake into RMS cells compared to unfunctionalized cubosomes. CD44 and CD221 directed magnetic (triple-function) cubosomes were capable of internalizing into RMS cells in an energy-independent mechanism. Helenalin-laden triple functionalized cubosomes showed limited impact on the viability of control fibroblast cells, while they induced a high degree cytotoxicity against RMS cells. Profound tumour cell death was observed in both two-dimensional (2D) culture and three-dimensional (3D) tumour spheroids.

Recent clinical cohort studies have highlighted that there is a three-fold greater SARS-Cov-2 infection risk in cancer patients, and overall mortality in individuals with tumours is increased by 41% with respect to general COVID-19 patients. Thus, access to therapeutics and intensive care is compromised for people with both diseases (comorbidity) and there is risk of delayed access to diagnosis. This comorbidity has resulted in extensive burden on the treatment of patients and health care system across the globe; moreover, mortality of hospitalized patients with comorbidity is reported to be 30% higher than for individuals affected by either disease. In this data-driven review, we aim specifically to address drug discoveries and clinical data of cancer management during the COVID-19 pandemic. The review will extensively address the treatment of COVID-19/cancer comorbidity; treatment protocols and new drug discoveries, including the description of drugs currently available in clinical settings; demographic features; and COVID-19 outcomes in cancer patients worldwide.

'Bioinks' are important tools for the fabrication of artificial living-tissue constructs that are able to mimic all properties of native tissues via 3D bioprinting technologies. Bioinks are most commonly made by incorporating live cells of interest within a natural or synthetic biocompatible polymeric matrix. In oncology research, the ability to recreate a tumor microenvironment (TME) using by 3D bioprinting constitutes a promising approach for drug development, screening, and in vitro cancer modeling. Here, we review the different types of bioink used for 3D bioprinting, with a focus on its application in cancer management. In addition, we consider the fabrication of bioink using customized materials/cells and their properties in the field of cancer drug discovery.

The aim of this Special Issue, “Nanoparticles for cancer therapy”, was to offer readers a comprehensive and up-to-date insight into the various applications of nanoparticles in cancer treatments [...]

Nanomaterials have outstanding applications in biomedical areas, such as drug delivery agents, nanoparticles-based imaging, magnetic fluid hyperthermia, and tissue regeneration. In the line of extensive development in nanomaterials-aided drug delivery platforms, pharmacokinetic assessments have achieved greater consideration from scientists and pharmacists as a fundamental component of the study of these nanosystems. Pharmacokinetics is nothing but the study of liberation, absorption, distribution, metabolization, and excretion (i.e., LADME) of the particular drug at the molecule scale, which provides significant information about drug response and associated side effects. The utilization of nano platforms in the delivery of drugs explores new possibilities to deliver drugs in the form of particulates, which specifically alters the LADME profile. The proper information of the pharmacokinetics aspects associated with the LADME plays a vital role in designing appropriate nanomaterials-based drug delivery systems for final clinical assessments. The pharmacokinetics of the nanomaterials is primarily dependent on the adsorption of proteins, which is consecutively decided by the physicochemical properties of the nanomaterials. Thus, more insights into the interrelation between the physicochemical properties of nanomaterials and their interface with body is required to recognize and envisage the in vivo response of the nanomaterials. In this chapter, we highlight some of the recent insights of the pharmacokinetic investigations of nanomaterials. We also provide the correlation between the physicochemical properties of the nanoscale materials and their biological behavior.

Rhabdomyosarcoma (RMS) is the most frequent soft tissue sarcoma in paediatric patients. Relapsed or refractory RMS shows very low 5-year survival rates, which urgently necessitates new chemotherapy agents. Herein, the sesquiterpene lactone, helenalin, was investigated as a new potential therapeutic agent against the embryonal RMS (eRMS) and alveolar RMS (aRMS) cells. We have evaluated in vitro antiproliferative efficacy of helenalin on RMS cells by the MTT and wound healing assay, and estimated several cell death pathways by flow cytometry, confocal microscopy and immunoblotting. It was shown that helenalin was able to increase reactive oxygen species levels, decrease mitochondrial membrane potential, trigger endoplasmic reticulum stress and deactivate the NF-κB pathway. Confirmation was obtained through the use of antagonistic compounds which alleviated the effects of helenalin in the corresponding pathways. Our findings demonstrate that oxidative stress is the pivotal mechanism of action of helenalin in promoting RMS cell death in vitro.

Rhabdomyosarcoma (RMS) is a rare type of soft tissue sarcoma most commonly found in pediatric patients. Despite progress, new and improved drug regimens are needed to increase survival rates. Citral, a natural product plant oil can induce cell death in cancer cells. Another compound, metformin, isolated originally from French lilac and used by diabetics, has been shown to reduce the incidence of cancer in these patients. Application of citral to RMS cells showed increase in cell death, and RD and RH30 cells showed half maximal inhibitory concentration (IC50) values as low as 36.28 μM and 62.37 μM, respectively. It was also shown that the citral initiated cell apoptosis through an increase in reactive oxygen species (ROS) and free calcium. In comparison, metformin only showed moderate cell death in RMS cell lines at a very high concentration (1,000 μM). Combinatorial experiments, however, indicated that citral and metformin worked antagonistically when used together. In particular, the ability of metformin to quench the ROS induced by citral could lead to the suppression of activity. These results clearly indicate that while clinical use of citral is a promising anti‐tumor therapy, caution should be exercised in patients using metformin for diabetes.

Plant volatile organic compounds (volatiles) are secondary plant metabolites that play crucial roles in the reproduction, defence, and interactions with other vegetation. They have been shown to exhibit a broad range of biological properties and have been investigated for antimicrobial and anticancer activities. In addition, they are thought be more environmentally friendly than many other synthetic chemicals 1 . Despite these facts, their applications in the medical, food, and agricultural fields are considerably restricted due to their volatilities, instabilities, and aqueous insolubilities. Nanoparticle encapsulation of plant volatile organic compounds is regarded as one of the best strategies that could lead to the enhancement of the bioavailability and biological activity of the volatile compounds by overcoming their physical limitations and promoting their controlled release and cellular absorption. In this review, we will discuss the biosynthesis and analysis of plant volatile organic compounds, their biological activities, and limitations. Furthermore, different types of nanoparticle platforms used to encapsulate the volatiles and the biological efficacies of nanoencapsulated volatile organic compounds will be covered.

A type of chromosome-free cell called SimCells (simple cells) has been generated from Escherichia coli, Pseudomonas putida, and Ralstonia eutropha. The removal of the native chromosomes of these bacteria was achieved by double-stranded breaks made by heterologous I-CeuI endonuclease and the degradation activity of endogenous nucleases. We have shown that the cellular machinery remained functional in these chromosome-free SimCells and was able to process various genetic circuits. This includes the glycolysis pathway (composed of 10 genes) and inducible genetic circuits. It was found that the glycolysis pathway significantly extended longevity of SimCells due to its ability to regenerate ATP and NADH/NADPH. The SimCells were able to continuously express synthetic genetic circuits for 10 d after chromosome removal. As a proof of principle, we demonstrated that SimCells can be used as a safe agent (as they cannot replicate) for bacterial therapy. SimCells were used to synthesize catechol (a potent anticancer drug) from salicylic acid to inhibit lung, brain, and soft-tissue cancer cells. SimCells represent a simplified synthetic biology chassis that can be programmed to manufacture and deliver products safely without interference from the host genome.

Biogenic silver nanoparticles are used for a number of applications due to their size, surface characteristics and strong antimicrobial properties. The present study aimed to investigate the synthesis of silver nanoparticles from the indigenous bacterial strain Bacillus sp. MB353 (PRJNA357966). Detailed characterization of silver nano particles was performed by UV-vis Spectrophotometer, FTIR, SEM and XRD. Biogenic silver nanoparticles were crystalline with average size 49-53 nm. These silver nanoparticles demonstrated good antibacterial activity against Gram-positive and Gram-negative bacteria (E. coli, Bacillus subtilis, Staphylococcus aureus, Klebsiella pneumoniae, Enterococcus faecium, Enterococcus faecalis and Streptomyces laurentii). The nanoparticles also showed excellent antifungal activity against Aspergillus niger, Aspergillus fumigatus, Fusarium soleni. The silver nano particles showed negligible antioxidant activity, but ROS generation pointed to a possible mode of antimicrobial activity. Incubation of silver nanoparticles with mammalian cell lines (Rhabdomyosarcomas and fibroblast) showed cell death and inhibition of proliferation. Cytotoxicity was most likely a result of ROS generation and changes in intracellular calcium levels. These findings suggested that biogenic silver nanoparticles could be used as alternative agents for biomedical purposes such as antibacterial and antifungal agents. However, given the effects on normal mammalian cells, it is probable that they could be used as anticancer agents if applied in targeted therapies.

To date, various chemically synthesized and biosynthesized nanoparticles, or hybrid nanosystems and/or nanoplatforms, have been developed under the umbrella of nanomedicine. These can be introduced into the body orally, nasally, intratumorally or intravenously. Successfully translating hybrid nanoplatforms from preclinical proof-of-concept to therapeutic value in the clinic is challenging. Having made significant advances with drug delivery technologies, we must learn from other areas of oncology drug development, where patient stratification and target-driven design have improved patient outcomes. This review aims to identify gaps in our understanding of the current strengths of nanomedicine platforms in drug delivery and cancer theranostics. We report on the current approaches of nanomedicine at preclinical and clinical stages.

Nanoparticles may be used in vaccinology as an antigen delivery and/or an immunostimulant to enhance immunity. Porous silica has been identified as an effective adjuvant for more than a decade, and we have therefore investigated the take up rate by an immortalized macrophage-like cell line of a number of mesoporous silica nanoparticles (MSNPs) with differing diameter and pore size. The MSNPs were synthesized using a sol-gel reaction and post-synthesis removal of the template. The MSNPs showed a clear distribution in take up rate peaking at 217 nm, whereas a comparison with solid spherical nanoparticles showed a similar distribution peaking at 377 nm. The MSNPs were investigated before and after loading with antigen. Diphtheria toxoid was used as a proof-of-concept antigen and showed a peak macrophage internalization of 53.42% for loaded LP3 particles which had a diameter of 217.75 ± 5.44 nm and large 16.5 nm pores. Optimal MSNP sizes appeared to be in the 200–400 nm range, and larger pores showed better antigen loading. The mesoporous silica particles were shown to be generally biocompatible, and cell viability was not altered by the loading of particles with or without antigen.