Microspheres loaded with polysaccharide nanoparticles for pulmonary delivery: preparation, structure and surface analysis

In this work, we report the preparation of a nanoparticle-based dry powder for pulmonary administration. Hybrid chitosan/hyaluronic acid nanoparticles were produced by ionotropic gelation and characterized for their physicochemical properties, being further studied by solid nuclear magnetic resonance (NMR). Using mannitol as carrier, nanoparticles were microencapsulated by spray drying, resulting in a dry powder with appropriate aerodynamic properties for lung delivery. In order to investigate the nanoparticles distribution within the carrier matrix, several techniques were applied that permitted an in-depth analysis of the system structure and surface, such as confocal laser scanning microscopy (CLSM) and X-ray photoelectron spectroscopy (XPS) in combination with time-of-flight secondary ion mass spectroscopy (TOF-SIMS). Overall, the studies conducted revealed that nanoparticles are homogeneously distributed through mannitol microspheres, suggesting the success of the microencapsulation process. In the light of these findings, it was concluded that the developed delivery system holds great potential for lung delivery of macromolecules

The potential of chitosan for pulmonary drug delivery

The administration of drugs through the pulmonary route offers great advantages, but also requires overcoming many challenges. There is a need to develop appropriate carriers for each active molecule to be delivered to the desired site in the lung, either for a local or a systemic effect. The polysaccharide chitosan is a very promising material for this purpose, given its demonstrated properties of biodegradability and biocompatibility, as well as mucoadhesivity and ability to enhance macromolecules permeation. In this review, the potential of chitosan to develop drug carriers for delivery to the lung will be discussed. The most important features that can support its selection will be explained. Besides, different approaches to increase its performance, especially concerning solubility, permeation-enhancing properties and gene transfection efficiency, will be presented. Special emphasis will be placed on information of different chitosan-based carriers, namely nanoparticles and microparticles, intended for pulmonary drug delivery

Mucosal delivery of liposome-chitosan nanoparticles complexes

Designing adequate drug carriers has long been a major challenge for those working in drug delivery. Since drug delivery strategies have evolved for mucosal delivery as the outstanding alternative to parenteral administration, many new drug delivery systems have been developed which evidence promising properties to address specific issues. Colloidal carriers, such as nanoparticles and liposomes, have been referred to as the most valuable approaches, but still have some limitations that can become more inconvenient as a function of the specific characteristics of administration routes. To overcome these limitations, we developed a new drug delivery system that results from the combination of chitosan nanoparticles and liposomes, in an approach of combining their advantages, while avoiding their individual limitations. These lipid/chitosan nanoparticle complexes are, thus, expected to protect the encapsulated drug from harsh environmental conditions, while concomitantly providing its controlled release. To prepare these assemblies, two different strategies have been applied: one focusing on the simple hydration of a previously formed dry lipid film with a suspension of chitosan nanoparticles, and the other relying on the lyophilization of both basic structures (nanoparticles and liposomes) with a subsequent step of hydration with water. The developed systems are able to provide a controlled release of the encapsulated model peptide, insulin, evidencing release profiles that are dependent on their lipid composition. Moreover, satisfactory in vivo results have been obtained, confirming the potential of these newly developed drug delivery systems as drug carriers through distinct mucosal routes

Rifabutin-Loaded Nanostructured Lipid Carriers as a Tool in Oral Anti-Mycobacterial Treatment of Crohn s Disease

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).Oral anti-mycobacterial treatment of Crohn’s disease (CD) is limited by the low aqueous solubility of drugs, along with the altered gut conditions of patients, making uncommon their clinical use. Hence, the aim of the present work is focused on the in vitro evaluation of rifabutin (RFB)-loaded Nanostructured lipid carriers (NLC), in order to solve limitations associated to this therapeutic approach. RFB-loaded NLC were prepared by hot homogenization and characterized in terms of size, polydispersity, surface charge, morphology, thermal stability, and drug payload and release. Permeability across Caco-2 cell monolayers and cytotoxicity and uptake in human macrophages was also determined. NLC obtained were nano-sized, monodisperse, negatively charged, and spheroidal-shaped, showing a suitable drug payload and thermal stability. Furthermore, the permeability profile, macrophage uptake and selective intracellular release of RFB-loaded NLC, guarantee an effective drug dose administration to cells. Outcomes suggest that rifabutin-loaded NLC constitute a promising strategy to improve oral anti-mycobacterial therapy in Crohn’s disease.This research was funded by V-A POCTEP Program (0245_IBEROS_1_E) of EU (FEDER), Xunta de Galicia (Competitive Reference Groups, ED431C 2016/008 and ED431C2017/09-FEDER), as well as by Fundação para a Ciência e Tecnologia, Portugal (under iMED.ULisboa project Pest-UID/DTP/04138/2019).info:eu-repo/semantics/publishedVersio

Rifabutin-Loaded Nanostructured Lipid Carriers as a Tool in Oral Anti-Mycobacterial Treatment of Crohn’s Disease

Oral anti-mycobacterial treatment of Crohn’s disease (CD) is limited by the low aqueous solubility of drugs, along with the altered gut conditions of patients, making uncommon their clinical use. Hence, the aim of the present work is focused on the in vitro evaluation of rifabutin (RFB)-loaded Nanostructured lipid carriers (NLC), in order to solve limitations associated to this therapeutic approach. RFB-loaded NLC were prepared by hot homogenization and characterized in terms of size, polydispersity, surface charge, morphology, thermal stability, and drug payload and release. Permeability across Caco-2 cell monolayers and cytotoxicity and uptake in human macrophages was also determined. NLC obtained were nano-sized, monodisperse, negatively charged, and spheroidal-shaped, showing a suitable drug payload and thermal stability. Furthermore, the permeability profile, macrophage uptake and selective intracellular release of RFB-loaded NLC, guarantee an effective drug dose administration to cells. Outcomes suggest that rifabutin-loaded NLC constitute a promising strategy to improve oral anti-mycobacterial therapy in Crohn’s diseaseThis research was funded by V-A POCTEP Program (0245_IBEROS_1_E) of EU (FEDER), Xunta de Galicia (Competitive Reference Groups, ED431C 2016/008 and ED431C2017/09-FEDER), as well as by Fundação para a Ciência e Tecnologia, Portugal (under iMED.ULisboa project Pest-UID/DTP/04138/2019)S

Pullulan-based nanoparticles as carriers for transmucosal protein delivery

Polymeric nanoparticles have revealed very effective in transmucosal delivery of proteins. Polysaccharides are among the most used materials for the production of these carriers, owing to their structural flexibility and propensity to evidence biocompatibility and biodegradability. In parallel, there is a preference for the use of mild methods for their production, in order to prevent protein degradation, ensure lower costs and easier procedures that enable scaling up. In this work we propose the production of pullulan-based nanoparticles by a mild method of polyelectrolyte complexation. As pullulan is a neutral polysaccharide, sulfated and aminated derivatives of the polymer were synthesized to provide pullulan with a charge. These derivatives were then complexed with chitosan and carrageenan, respectively, to produce the nanocarriers. Positively charged nanoparticles of 180-270 nm were obtained, evidencing ability to associate bovine serum albumin, which was selected as model protein. In PBS pH 7.4, pullulan-based nanoparticles were found to have a burst release of 30% of the protein, which maintained up to 24h. Nanoparticle size and zeta potential were preserved upon freeze-drying in the presence of appropriate cryoprotectants. A factorial design was approached to assess the cytotoxicity of raw materials and nanoparticles by the metabolic test MTT. Nanoparticles demonstrated to not cause overt toxicity in a respiratory cell model (Calu-3). Pullulan has, thus, demonstrated to hold potential for the production of nanoparticles with an application in protein delivery

Design and in vitro assessment of chitosan nanocapsules for the pulmonary delivery of rifabutin

Tuberculosis (TB) is a life-threatening disease and a main cause of death worldwide. It mainly affects the lungs, and it is attributed to the infection with Mycobacterium tuberculosis (MTB). Current treatments consist of the oral administration of combinations of antibiotics including rifabutin, in high doses and for long periods of time. These therapeutic regimens are associated with many side effects and high rates of drug resistance. To overcome these problems, this study aims at developing a nanosystem for the improved delivery of antibiotics, with potential application in pulmonary delivery. Chitosan-based nanomaterials are widely used in biomedical applications, due to their biodegradability and biocompatibility, as well as their potential antimicrobial effects and lack of toxicity. In addition, this polymer is particularly attractive for mucosal delivery due to its bioadhesive properties. Therefore, the structure of the proposed nanocarrier consists of a chitosan shell and a lipid core with a combination of different oils and surfactants to allow optimal association of the hydrophobic drug rifabutin. These nanocapsules were characterized in terms of size, polydispersity index, surface charge, morphology, encapsulation efficiency and biological stability. The release kinetics of the drug-loaded nanostructures was evaluated in simulated lung media. Moreover, in vitro studies in different cell models (A549 and Raw 264.7 cells) demonstrated the safety of the nanocapsules as well as their efficient internalization. An antimicrobial susceptibility test was performed to evaluate the efficacy of the rifabutin-loaded nanocapsules against Mycobacterium phlei. This study indicated complete inhibition for antibiotic concentrations within the expected susceptibility range of Mycobacterium (≤ 0.25–16 mg/L)This work has received financial support from the Xunta de Galicia (Centro singular de investigación de Galicia, accreditation 2019–2022); Competitive Reference Groups, ED431C 2021/17-FEDER) and Ministerio de Ciencia e Innovación, Gobierno de España (PID2019-107500RB-I00)S

A Traffic Light System to Maximize Carbohydrate Cryoprotectants’ Effectivity in Nanostructured Lipid Carriers’ Lyophilization

Lyophilization is often employed to transform nanoparticle suspensions to stable solid forms. This work proposed Neurofuzzy Logic (NFL) to better understand the lyophilization process of Nanostructured Lipid Carriers’ (NLCs) dispersions and the carbohydrate cryoprotectants’ (CPs) performance in these processes. NLCs were produced by hot homogenization, frozen at different speeds, and lyophilized using several CPs at variable concentrations. NLCs were characterized, and results were expressed as increase in particle size (Δ size), polydispersity (Δ PdI), and zeta potential (Δ ZP) of lyophilized powders (LP) regarding initial dispersions. CPs were classified according to their molecular weights (MW), and the osmolarities (Π) of CPs solutions were also determined. Databases obtained were finally modelled through FormRules® (Intelligensys Ltd., Kirkwall, Scotland, UK), an NFL software. NFL models revealed that CPs’ MW determines the optimal freezing conditions and CPs’ proportions. The knowledge generated allowed the establishment of a traffic light system intended to successfully select and apply sugars for nanoparticles lyophilization

Microencapsulated Chitosan-Based Nanocapsules: A New Platform for Pulmonary Gene Delivery

In this work, we propose chitosan (CS)-based nanocapsules (NCs) for pulmonary gene delivery. Hyaluronic acid (HA) was incorporated in the NCs composition (HA/CS NCs) aiming to promote gene transfection in the lung epithelium. NCs were loaded with a model plasmid (pCMV-βGal) to easily evaluate their transfection capacity. The plasmid encapsulation efficiencies were of approx. 90%. To facilitate their administration to the lungs, the plasmid-loaded NCs were microencapsulated in mannitol (Ma) microspheres (MS) using a simple spray-drying technique, obtaining dry powders of adequate properties. In vivo, the MS reached the deep lung, where the plasmid-loaded CS-based NCs were released and transfected the alveolar cells more homogeneously than the control formulation of plasmid directly microencapsulated in Ma MS. The HA-containing formulation achieved the highest transfection efficiency, in a more extended area and more homogeneously distributed than the rest of tested formulations. The new micro-nanostructured platform proposed in this work represents an efficient strategy for the delivery of genetic material to the lung, with great potential for the treatment of genetic lung diseases