Lipid-Based Nano-Sized Cargos as a Promising Strategy in Bone Complications : A Review

Funding Information: Funding: This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the grant agreement No 857287. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.Bone metastasis has been considered the fatal phase of cancers, which remains incurable and to be a challenge due to the non-availability of the ideal treatment strategy. Unlike bone cancer, bone metastasis involves the spreading of the tumor cells to the bones from different origins. Bone metastasis generally originates from breast and prostate cancers. The possibility of bone metastasis is highly attributable to its physiological milieu susceptible to tumor growth. The treatment of bone-related diseases has multiple complications, including bone breakage, reduced quality of life, spinal cord or nerve compression, and pain. However, anticancer active agents have failed to maintain desired therapeutic concentrations at the target site; hence, uptake of the drug takes place at a non-target site responsible for the toxicity at the cellular level. Interestingly, lipid-based drug delivery systems have become the center of interest for researchers, thanks to their biocompatible and biomimetic nature. These systems possess a great potential to improve precise bone targeting without affecting healthy tissues. The lipid nano-sized systems are not only limited to delivering active agents but also genes/peptide sequences/siRNA, bisphosphonates, etc. Additionally, lipid coating of inorganic nanomaterials such as calcium phosphate is an effective approach against uncontrollable rapid precipitation resulting in reduced colloidal stability and dispersity. This review summarizes the numerous aspects, including development, design, possible applications, challenges, and future perspective of lipid nano-transporters, namely liposomes, exosomes, solid lipid nanoparticles (SLN), nanostructured lipid carriers (NLC), and lipid nanoparticulate gels to treat bone metastasis and induce bone regeneration. Additionally, the economic suitability of these systems has been discussed and different alternatives have been discussed. All in all, through this review we will try to understand how far nanomedicine is from clinical and industrial applications in bone metastasis.publishersversionPeer reviewe

A Compiled Update on Nutrition, Phytochemicals, Processing Effects, Analytical Testing and Health Effects of Chenopodium album : A Non-Conventional Edible Plant (NCEP)

Funding Information: The authors acknowledge financial support from the European Union’s Horizon 2020 research and innovation programme under the grant agreement No. 857287 (BBCE—Baltic Biomaterials Centre of Excellence). Publisher Copyright: © 2023 by the authors.Bathua (Chenopodium album) is a rich source of extensive-ranging nutrients, including bio-active carbohydrates, flavonoids and phenolics, minerals, and vitamins that translate to countless health benefits such as anticancer, antidiabetic, anti-inflammatory, antimicrobial, and antioxidant activity. Ascaridole, an important phytoconstituent present in aerial parts of the plant, contributes to its anthelmintic property. Even with vast historical use and significant health benefits, its renown has not spread, and utilization has significantly decreased in recent decades. Gradually, the plant has become known under the name of Non-conventional edible plant (NCEP). This compilation is prepared to bring out the plant under the spotlight for further research by foregrounding previous studies on the plant. Scientific research databases, including PubMed, Google Scholar, Scopus, SpringerLink, ScienceDirect, and Wiley Online, were used to fetch data on C. album. This review offers over up-to-date knowledge on nutritious values, phytochemical composition, volatile compounds, as well as health benefits of C. album. The ethnobotanical and ethnomedicinal uses of the plant in India and other parts of the world are deliberately discussed. Scrutinizing the reported literature on C. album reveals its powerful nutrient composition advantageous in the development of food products. The impact of various cooking and processing methods on the nutritional profile and bioavailability are discussed. The future perspectives with regards to the potential for food and nutraceutical products are critically addressed. This review proves the necessity of breakthrough research to investigate the pharmacology and safety of phytochemicals and nutraceutical development studies on the C. album.Peer reviewe

Ellagic acid containing Nanostructured Lipid Carriers for topical application: a preliminary study

Ellagic acid (EA) is a potent antioxidant substance of natural origin characterized by poor biopharmaceutical properties and low solubility in water that limit their use. The aim of the present study was to develop lipid based nanoparticle formulations able to encapsulate EA for dermal delivery purpose. The EA-loaded nanoparticles were prepared using two different lipid compositions, namely tristearin/tricaprylin (NLC-EA1) and tristearin/labrasol (NLC-EA2). The influence of formulations on size, entrapment efficiency and stability of EA-loaded nanoparticles was investigated. Cryo-TEM and SAXS analysess showed that no morphological differences are evident among all the types of loaded and unloaded NLC. The macroscopic aspect of both NLC-EA1 and NLC-EA2 did not change by time. No difference in size is appreciable between empty and drug-containing NLC, thus the nanoparticle diameter is not affected by the presence EA and in general no variations of the diameters occur during time. The percentage of entrapment efficiency of both EA-loaded nanoparticles was almost quantitative. In addition NLC-EA1 maintain EA stability for almost 2 months, while NLC-EA2 up to 40 days. FRAP assay showed an antioxidant activity around 60% for both the loaded NLC, as compared to the solution. Although both types of NLC are characterized by some toxicity, NLC-EA1 are less cytotoxic than NLC-EA2. Taken together these results demonstrated that the inclusion of EA within NLC could improve the water solubility, allowing for a reduction of the dosage. Moreover, the maintaining of high antioxidant effect and low toxicity were evidenced for both types of NLC-EA

Ethosomes and transethosomes for mangiferin transdermal delivery

Mangiferin is a natural glucosyl xanthone with antioxidant and anti-inflammatory activity, making it suitable for protection against cutaneous diseases. In this study ethosomes and transethosomes were designed as topical delivery systems for mangiferin. A preformulation study was conducted using different surfactants in association with phosphatidylcholine. Vesicle dimensional distribution was monitored by photon correlation spectroscopy, while antioxidant capacity and cytotoxicity were respectively assessed by free radical scavenging analysis and MTT on HaCaT keratinocytes. Selected nanosystems were further investigated by cryogenic transmission electron microscopy, while mangiferin entrapment capacity was evaluated by ultracentrifugation and HPLC. The diffusion kinetics of mangiferin from ethosomes and transethosomes evaluated by Franz cell was faster in the case of transethosomes. The suitability of mangiferin-containing nanovesicles in the treatment of skin disorders related to pollutants was investigated, evaluating, in vitro, the antioxidant and anti-inflammatory effect of ethosomes and transethosomes on human keratinocytes exposed to cigarette smoke as an oxidative and inflammatory challenger. The ability to induce an antioxidant response (HO-1) and anti-inflammatory status (IL-6 and NF-kB) was determined by RT-PCR and immunofluorescence. The data demonstrated the effectiveness of mangiferin loaded in nanosystems to protect cells from damage. Finally, to gain insight into the keratinocytes' uptake of ethosome and transethosome, transmission electron microscopy analyses were conducted, showing that both nanosystems were able to pass intact within the cells

Design strategies and technical aspects of nanoscale drug carriers

SCOPO L'obiettivo principale di questa ricerca si basa sullo sviluppo e la caratterizzazione di formulazioni a base di lipidi, ovvero nanoparticelle magnetiche rivestite di lipidi (MNP) per applicazioni teranostiche, nanoparticelle lipidiche solide (SLN), carriers lipidici nanostrutturati (NLC), etosomi (ETHO), transetosomi, gel per il rilascio di farmaci dermico e transdermico e liposomi per il trasporto di inibitori del biofilm. La tesi si è concentrata principalmente sugli aspetti riguardanti il rilascio di alcune molecole antiossidanti (come acido ellagico, acido caffeico, mangiferina, curcumina) per applicazioni cutanee. Attraverso la scelta dell’opportuna composizione del nanosistema è stato così possibile affrontare e risolvere i problemi di scarsa solubilità di queste molecole. Inoltre, è stato proposto uno studio preliminare di formulazione liposomiale per la somministrazione di inibitori sintetici del quorum sensing, finalizzata a risolvere il problema della resistenza agli antibiotici in particolari patologie. Infine è stato condotto uno studio formulativo per la inclusione di nanoparticelle magnetiche in nanosistemi lipidici finalizzata alla riduzione della loro tossicità in seguito a somministrazione. METODI La morfologia e la distribuzione delle dimensioni dei nanosistemi caricati con le diverse tipologie di farmaco sono state studiate mediante microscopia elettronica, mentre la loro struttura interna è stata studiata utilizzando SAXS. I diametri medi sono stati valutati con misure PCS. La tecnica HPLC è stata impiegata per valutare l'efficienza dell'incapsulamento e la stabilità chimica dei nanosistemi prodotti. Il potere antiossidante è stato valutato con metodo DPPH (2,2-difenil-1-picrylhydrazyl). L'influenza del tipo di nanosistema sulla diffusione delle molecole incluse è stata valutata utilizzando la cella di Franz associata ad una membrana di nylon, mentre la permeazione attraverso la pelle è stato valutata attraverso uno studio amperometrico, utilizzando un elettrodo SCOE (skin covered oxygen electrode) ricoperto con pelle di suino. Inoltre è stato valutato l'effetto delle formulazioni sulla resistenza e l'integrità della pelle. Sono state impiegate varie tecniche in-vitro ed ex-vivo, quali saggi di citotossicità, d’integrità del biofilm, di esposizione al fumo di sigaretta e analisi western blot. RISULTATI I risultati principali ottenuti sono rappresentati da un miglioramento della solubilità e della stabilità nel tempo delle molecole incapsulate. È stato infatti dimostrato che l'inclusione dei farmaci modello all'interno dei nanosistemi potrebbe migliorarne la solubilità in acqua consentendo una riduzione del dosaggio. In particolare, lo studio e i risultati ottenuti con l'elettrodo SCOE rivelano i limiti ma anche le possibilità nello sviluppo di strategie innovative e versatili per comprendere l'efficacia dei nanosistemi come vettori per composti fenolici. Questo approccio permetterebbe inoltre di capire se l'applicazione topica possa avere anche un effetto protettivo non solo sull'epidermide superiore ma anche sugli strati cutanei profondi, come strato basale e derma. In secondo luogo, i liposomi cationici potrebbero essere considerati un promettente sistema di rilascio per gli inibitori del quorum sensing e possono interrompere la formazione di biofilm per migliorare l'efficacia dell'antibiotico. CONCLUSIONI Sebbene la nanomedicina sia ancora in fase di sviluppo, molti agenti attivi terapeutici che sfruttano la nanotecnologia sono stati approvati e commercializzati. Tuttavia, affinché si possano compiere progressi significativi verso questo obiettivo, sono necessari ulteriori sforzi per stabilire criteri di test, convalidare l'efficacia e accumulare dati di sicurezza per l’utilizzo di questi innovativi agenti e materiali nanoterapeutici.PURPOSE The main aim of this research is based on the development and characterization of lipid-based formulations namely lipid-coated magnetic nanoparticles (MNP) for theranostic applications, Solid lipid nanoparticles (SLN), Nanostructured lipid carriers (NLC), ethosomes (ETHO), transethosomes, gels for dermal and transdermal drug delivery, and liposomes to target biofilm inhibitors. The thesis has mainly focussed on a wide range of aspects such as the delivery of anti-oxidant molecules (ellagic acid, caffeic acid, mangiferin, curcumin) for cutaneous applications. Wherein, their poor solubility problems have been addressed and solved by choosing different compositions and nano-systems. Moreover, a preliminary study on liposomal delivery of synthetic quorum sensing inhibitors has been proposed, which can be helpful to solve the issue of antibiotic resistance. Finally, the last study was to modify the toxicity associated with magnetic nanoparticles by incorporating them into lipid nanoparticles. METHODS The morphology and size distribution of nanosystems loaded with different drug molecules have been investigated by electron microscopy and internal structure using SAXS. The mean diameters have been evaluated by PCS. The HPLC techniques have been employed to assess encapsulation efficiency and chemical stability. The antioxidant power has been evaluated by the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) methodology. The influence of the type of nanosystems on the diffusion of drug molecules has been evaluated by Franz cell-associated to a nylon membrane, while to evaluate their permeation through the skin, an amperometric study has been conducted, based on porcine SCOE. The effect of formulations on skin resistance and integrity has been evaluated. The various in-vitro and ex-vivo techniques including cytotoxicity assays, biofilm assays, and cigarette smoke exposure, western blot analysis respectively have been employed. RESULTS Firstly, the improved solubility and shelf life of drug molecules have been achieved. It was demonstrated that the inclusion of drug molecules within nanosystems could improve the water solubility, allowing for a reduction of the dosage. Notably, the work and results obtained with skin or enzymes modified oxygen electrode reveal the limitations and the possibilities for developing novel and versatile strategies to understand the effectiveness of nanosystems as carriers for phenolic compounds. This would allow understanding whether the topical application could have even a protective effect not only in the upper epidermis but also even at the lower cutaneous layers, such as basal epidermis and dermis. Secondly, cationic liposomes could be considered as a promising delivery system for quorum sensing inhibitors and can interrupt the biofilms formation to enhance antibiotic efficacy. CONCLUSIONS Though nanomedicine is still at an initial phase of development, many therapeutic active agents that exploit nanotechnology have been accepted and commercialized. However, for significant progress to be made toward this goal, much more effort is needed to establish testing criteria, validate efficacy, and accumulate safety data for various nanotherapeutic agents and materials

The Potential of Caffeic Acid Lipid Nanoparticulate Systems for Skin Application : In Vitro Assays to Assess Delivery and Antioxidant Effect

The object of this study is a comparison between solid lipid nanoparticles and ethosomes for caffeic acid delivery through the skin. Caffeic acid is a potent antioxidant molecule whose cutaneous administration is hampered by its low solubility and scarce stability. In order to improve its therapeutic potential, caffeic acid has been encapsulated within solid lipid nanoparticles and ethosomes. The effect of lipid matrix has been evaluated on the morphology and size distribution of solid lipid nanoparticles and ethosomes loaded with caffeic acid. Particularly, morphology has been investigated by cryogenic transmission electron microscopy and small angle X-ray scattering, while mean diameters have been evaluated by photon correlation spectroscopy. The antioxidant power has been evaluated by the 2,2-diphenyl-1-picrylhydrazyl methodology. The influence of the type of nanoparticulate system on caffeic acid diffusion has been evaluated by Franz cells associated to the nylon membrane, while to evaluate caffeic acid permeation through the skin, an amperometric study has been conducted, which was based on a porcine skin-covered oxygen electrode. This apparatus allows measuring the O2 concentration changes in the membrane induced by polyphenols and H2O2 reaction in the skin. The antioxidative reactions in the skin induced by caffeic acid administered by solid lipid nanoparticles or ethosomes have been evaluated. Franz cell results indicated that caffeic acid diffusion from ethosomes was 18-fold slower with respect to solid lipid nanoparticles. The amperometric method evidenced the transdermal delivery effect of ethosome, indicating an intense antioxidant activity of caffeic acid and a very low response in the case of SLN. Finally, an irritation patch test conducted on 20 human volunteers demonstrated that both ethosomes and solid lipid nanoparticles can be safely applied on the skin

Design of Nanosystems for the Delivery of Quorum Sensing Inhibitors : A Preliminary Study

Biofilm production is regulated by the Quorum Sensing system. Nowadays, Quorum Sensing represents an appealing target to design new compounds to increase antibiotics effects and avoid development of antibiotics multiresistance. In this research the use of liposomes to target two novel synthetic biofilm inhibitors is presented, focusing on a preformulation study to select a liposome composition for in vitro test. Five different liposome (LP) formulations, composed of phosphatidyl choline, cholesterol and charged surfactant (2:1:1, molar ratio) have been prepared by direct hydration and extrusion. As charged surfactants dicetyl phosphate didecyldimethylammonium chloride, di isobutyl phenoxy ethyl dimethyl benzyl ammonium chloride and stearylamine (SA) and have been used. Liposome charge, size and morphology were investigated by zeta potential, photon correlation spectroscopy, small angle x-ray spectroscopy and electron microscopy. LP-SA was selected for the loading of biofilm inhibitors and subjected to high performance liquid chromatography for entrapment capacity evaluation. LP-SA loaded inhibitors showed a higher diameter (223.6 nm) as compared to unloaded ones (205.7 nm) and a dose-dependent anti-biofilm effect mainly after 48 h of treatment, while free biofilm inhibitors loose activity. In conclusion, our data supported the use of liposomes as a strategy to enhance biofilm inhibitors effect

Design and Characterization of Ethosomes for Transdermal Delivery of Caffeic Acid.

in ethosome after six months, while in water, an almost complete degradation occurred within one month. The addition of poloxamer slightly modified vesicle structure and size, while it decreased the vesicle deformability. Caffeic acid diffusion coefficients from ethosome and ethosome gel were, respectively, 137- and 33-fold lower with respect to the aqueous solution. At last, the caffeic acid permeation and antioxidant power of ethosome were more intense with respect to the simple solution