Effect of experimental parameters on alginate/chitosan microparticles for BCG encapsulation

The aim of the present study was to develop novel Mycobacterium bovis bacille Calmette-Guérin (BCG)-loaded polymeric microparticles with optimized particle surface characteristics and biocompatibility, so that whole live attenuated bacteria could be further used for pre-exposure vaccination against Mycobacterium tuberculosis by the intranasal route. BCG was encapsulated in chitosan and alginate microparticles through three different polyionic complexation methods by high speed stirring. For comparison purposes, similar formulations were prepared with high shear homogenization and sonication. Additional optimization studies were conducted with polymers of different quality specifications in a wide range of pH values, and with three different cryoprotectors. Particle morphology, size distribution, encapsulation efficiency, surface charge, physicochemical properties and biocompatibility were assessed. Particles exhibited a micrometer size and a spherical morphology. Chitosan addition to BCG shifted the bacilli surface charge from negative zeta potential values to strongly positive ones. Chitosan of low molecular weight produced particle suspensions of lower size distribution and higher stability, allowing efficient BCG encapsulation and biocompatibility. Particle formulation consistency was improved when the availability of functional groups from alginate and chitosan was close to stoichiometric proportion. Thus, the herein described microparticulate system constitutes a promising strategy to deliver BCG vaccine by the intranasal route

In vitro cytocompatibility evaluation of poly(DL-lactic acid) scaffolds loaded with minocycline and voriconazole addressing osteomyelitis

Purpose: In this work is proposed the cytocompatibility evaluation of an innovative system based on the dual delivery of minocycline (Min) and voriconazole (Vor), aiming for bone infection therapeutics.info:eu-repo/semantics/publishedVersio

Chitosan and hyaluronic acid nanoparticles as vehicles of epoetin beta for subconjunctival ocular delivery

Research Areas: Pharmacology & PharmacyNeuroprotection in glaucoma using epoetin beta (EPOβ) has yielded promising results. Our team has developed chitosan-hyaluronic acid nanoparticles (CS/HA) designed to carry EPOβ into the ocular globe, improving the drug’s mucoadhesion and retention time on the ocular surface to increase its bioavailability. In the present in vivo study, we explored the possibility of delivering EPOβ to the eye through subconjunctival administration of chitosan-hyaluronic acid-EPOβ (CS/HA-EPOβ) nanoparticles. Healthy Wistar Hannover rats (n = 21) were split into 7 groups and underwent complete ophthalmological examinations, including electroretinography and microhematocrit evaluations before and after the subconjunctival administrations. CS/HA-EPOβ nanoparticles were administered to the right eye (OD), and the contralateral eye (OS) served as control. At selected timepoints, animals from each group (n = 3) were euthanized, and both eyes were enucleated for histological evaluation (immunofluorescence and HE). No adverse ocular signs, no changes in the microhematocrits (≈45%), and no deviations in the electroretinographies in both photopic and scotopic exams were observed after the administrations (p < 0.05). Intraocular pressure remained in the physiological range during the assays (11–22 mmHg). EPOβ was detected in the retina by immunofluorescence 12 h after the subconjunctival administration and remained detectable until day 21. We concluded that CS/HA nanoparticles could efficiently deliver EPOβ into the retina, and this alternative was considered biologically safe. This nanoformulation could be a promising tool for treating retinopathies, namely optic nerve degeneration associated with glaucoma.info:eu-repo/semantics/publishedVersio

Alginate-chitosan particulate delivery systems for mucosal immunization against tuberculosis

Although vaccination is still the most cost-effective strategy for tuberculosis control, there is an urgent need for an improved vaccine. Current BCG vaccine lacks efficacy in preventing adult pulmonary tuberculosis, the most prevalent form of the disease. Targeting nasal mucosa, Mycobacterium tuberculosis infection site, will allow a simpler, less prone to risk of infection and more effective immunization against disease. Due to its biodegradable, immunogenic and mucoadhesive properties, chitosan particulate delivery systems can act both as carrier and as adjuvant, improving the elicited immune response. In this study, BCG was encapsulated in alginate and chitosan microparticles, via a mild ionotropic gelation procedure with sodium tripolyphosphate as a counterion. The particulate system developed shows effective modulation of BCG surface physicochemical properties, suitable for mucosal immunization. Intracellular uptake was confirmed by effective transfection of human macrophage cell lines

Chitosan-alginate microparticulate delivery system for an alternative route of administration of BCG vaccine

Immunisation against M. tuberculosis with current available BCG vaccine lacks efficacy in preventing adult pulmonary tuberculosis. Targeting nasal mucosa is an attractive option for a more effective immunization. The delivery of BCG via the intranasal route involves overcoming barriers such as crossing the physical barrier imposed by the mucus layer and ciliar remotion, cellular uptake and intracellular trafficking by antigen presenting cells. Due to its biodegradable, immunogenic and mucoadhesive properties, chitosan particulate delivery systems can act both as vaccine carrier and adjuvant, improving the elicited immune response. In this study, different combinations of Chitosan/Alginate/TPP microparticles with BCG were produced as vaccine systems. The developed microparticle system successfully modulates BCG surface physicochemical properties and promotes effective intracellular uptake by human macrophage cell lines Preliminary immune responses were evaluated after s.c. and intranasal immunisation of BALB/c mice. BCG vaccination successfully stimulated the segregation of IgG2a and IgG1, where intranasal immunisation with chitosan/alginate particulate system efficiently elicited a more equilibrated cellular/humoral immune response

Infliximab microencapsulation: an innovative approach for intra-articular administration of biologics in the management of rheumatoid arthritis-in vitro evaluation

Microencapsulation of the therapeutical monoclonal antibody infliximab (INF) was investigated as an innovative approach to improve its stability and to achieve formulations with convenient features for intra-articular administration. Ultrasonic atomization (UA), a novel alternative to microencapsulate labile drugs, was compared with the conventional emulsion/evaporation method (Em/Ev) using biodegradable polymers, specifically Polyactive® 1000PEOT70PBT30 [poly(ethylene-oxide-terephthalate)/poly(butylene-terephthalate); PEOT-PBT] and its polymeric blends with poly-(D, L-lactide-co-glycolide) (PLGA) RG502 and RG503 (PEOT-PBT:PLGA; 65:35). Six different formulations of spherical core-shell microcapsules were successfully developed and characterized. The UA method achieved a significantly higher encapsulation efficiency (69.7-80.25%) than Em/Ev (17.3-23.0%). Mean particle size, strongly determined by the microencapsulation method and to a lesser extent by polymeric composition, ranged from 26.6 to 49.9 µm for UA and 1.5-2.1 µm for Em/Ev. All formulations demonstrated sustained INF release in vitro for up to 24 days, with release rates modulated by polymeric composition and microencapsulation technique. Both methods preserved INF biological activity, with microencapsulated INF showing higher efficacy than commercial formulations at comparable doses regarding bioactive tumor necrosis factor-alpha (TNF-α) neutralization according to WEHI-13VAR bioassay. Microparticles' biocompatibility and extensive internalization by THP-1-derived macrophages was demonstrated. Furthermore, high in vitro anti-inflammatory activity was achieved after treatment of THP-1 cells with INF-loaded microcapsules, significatively reducing in vitro production of TNF-α and interleucine-6 (Il-6)Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. Iván Lamela-Gómez received funding from the “Axudas á etapa predoutoral da Xunta de Galicia, cofinanciadas polo programa operativo FSE Galicia 2014–2020” predoctoral grant program. Consellería de Cultura, Educación e ordenación universitaria, Xunta de Galicia, Spain. This research was partially funded by Consellería de Cultura, Educación e ordenación universitaria, Xunta de Galicia, Spain. Axudas para a consolidación e estructuración de unidades de investigación competitivas Modalidad A: Grupos de Referencia Competitiva (ED341C 2017/13). This research was also partially funded by the Fundação para a Ciência e a Tecnologia (FCT), Portugal (UID/DTP/04138/2019 and UIDB/04138/2020 to iMed.ULisboa), and principal investigator grants CEECIND/03143/2017 (L. M. Gonçalves)S

Release kinetic model and antimicrobial activity of an innovative minocycline and voriconazole co-delivery system

Purpose and Strategy: Development of a new local drug-delivery system aiming at bone infection and the modulation of the polymicrobial activity; simultaneous delivery of minocycline and voriconazole, antibacterial and antifungal agents, respectively; polylactide (PDLLA) scaffolds functionalized with collagen and bioglass, osteogenic enhancers.info:eu-repo/semantics/publishedVersio

Poly(DL-lactic acid) scaffolds adsorbed with minocycline and voriconazole: a new pathway towards infection containment

Bone infection (osteomyelitis): burden as a clinical complication of orthopedic surgeries. Controlled antimicrobial release systems: treat and prevent osteomyelitis. Biomaterials based on porous scaffolds: local administration of high concentration of drugs; no systemic toxicity; extended time. Scaffolds in bone tissue engineering: a combination of bioresorbable polymers with bioactive bioglasses; present biodegradability and biosafety; suitable microenvironment and structure; favor osteogenic differentiation and cell growth. Co-encapsulation of drugs: advantageous means for administration of drugs; novel strategy directed to the co-delivery of two antimicrobials (voriconazole and minocycline).info:eu-repo/semantics/publishedVersio

Investigation of the genotoxicity of digested titanium dioxide nanomaterials in human intestinal cells

The widespread use of titanium dioxide nanomaterials (TiO2 NMs) in food and consumer products such as toothpaste or food contact materials, suggests the relevance of human oral exposure to these nanomaterials (NMs) and raises the possibility of adverse effects in the gastrointestinal tract (GIT). We previously showed that the in vitro digestion of TiO2 NMs may increase their toxicity in intestinal cells. In this work, we analyzed the genotoxicity and the intracellular reactive oxygen species induction by physiologically relevant concentrations of three different TiO2 NMs (NM-102, NM-103 and NM-105) in Caco-2 and HT29-MTX-E12 intestinal cells, while considering the potential influence of the digestion process in the NMs' physiochemical characteristics. The results evidenced a DNA-damaging effect dependent on the NM, more relevant for the rutile/anatase NM-105, possibly due to its lower hydrodynamic size in the cells medium. In addition, the results of the micronucleus assay suggest effects on chromosomal integrity, an indicator of cancer risk, in the HT29-MTX-E12 cells, for all the tested TiO2 NMs, especially after the in vitro digestion. This work supports the evidence for concerns on the use of TiO2 NMs as a food additive, recently reported by EFSA, and for their use in applications in consumer products that may drive human exposure through ingestion.This work was funded by national funds through the FCT - Foundation for Science and Technology, I.P., under the project PTDC/SAU-PUB/29481/2017. Research co-funded by UIDB/00009/2020; UIDP/00009/2020 (Centre for Toxicogenomics and Human Health – ToxOmics, FCT- Foundation for Science and Technology). NV holds a FCT PhD Scholarship grant (2020.07168.BD). iMed.ULisboa (UIDB/04138/2020 and UIDP/04138/2020) principal investigator grants CEECIND/03143/2017 (L. M. Gonçalves).info:eu-repo/semantics/publishedVersio

Multispectral and hyperspectral remote sensing as a source of knowledge in the Portuguese sector of the Iberian Pyrite Belt

ABSTRACT: Remote sensing is an invaluable tool to increase geological and mining knowledge, due to its screening view and variable discrimination and identification capabilities of the target materials. In this study an overview of remote sensing research developed and ongoing within the Portuguese sector of the Iberian Pyrite Belt (PSIPB) since 2000 is given. Multispectral and hyperspectral datasets were processed using hybrid methods, related both to general and detailed characterization, to: 1) support geological, mineral and hydrothermal mapping, 2) generate products derived from multivariate analysis and band ratios, 3) enhance correlation with radiometric data, 4) provide elements for environmental assessment concerning mining activity, 5) map Acid Mine Drainage (AMD) based on spectral field signatures, 6) quantify AMD based on high correlation mineralogical mapping, and 7) monitor AMD. The results highlight the importance of the quantitative digital support given by remote sensing tools within the Portuguese Sector of the Iberian Pyrite Belt (PSIPB), ruled by georesource exploitation in different stages of the Mine Lyfe Cycle.RESUMO: A deteção remota é uma ferramenta valiosa para aumentar o conhecimento geológico e mineiro, devido à visão sinótica e à capacidade variável de discriminação e identificação dos materiais-alvo. Neste trabalho dá-se uma visão geral da investigação através dos trabalhos de deteção remota desenvolvidos e em curso no Setor Português da Faixa Piritosa Ibérica (SPFPI) desde 2000. Os dados multiespectrais e hiperespectrais foram processados usando métodos híbridos quer para a sua caracterização geral quer detalhada para: 1) apoiar a cartografia geológica, de mineralizações e sistemas hidrotermais, 2) gerar produtos de análise multivariada e rácios de bandas, 3) melhorar a correlação com dados radiométricos 4) fornecer elementos para avaliação ambiental em áreas mineiras, 5) cartografar a drenagem ácida de mina (DAM) com assinaturas espectrais de campo, 6) quantificar a DAM através de cartografia mineralógica de alta correlação, e 7) monitorizar a DAM. Destaca-se a importância do suporte digital quantitativo dado por ferramentas de deteção remota no SPFPI, regido pela exploração de georrecursos em diferentes fases do Ciclo de Vida das Minas