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1.
Life Sci ; 264: 118502, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33031825

RESUMO

Bone tissue engineering compasses the use of mesenchymal stem cells (MSCs) along with engineered biomaterial construct to augment bone regeneration. Till now, MSCs were isolated from various sources and used in cellular constructs. For the first time, in this study, MSCs were isolated from human Ovarian Follicular Fluid (OFF) and characterized by CD 44+ and CD 105+ markers via confocal microscopy and flow cytometry. Additionally, MSCs stemness, proliferation and colony-forming unit ability, multi-lineage differentiation potential were also studied. To test its suitability for bone tissue engineering applications, we grew the MSCs with the conditioned medium obtained from biocomposite scaffold by fusing a natural polymer, Chitosan (CS) and a synthetic polymer, Polycaprolactone (PCL) and the scaffold were coated with Zinc divalent ions to impart osteogenic properties. The physico-chemical characterization of scaffold, such as FTIR, XRD, and SEM studies was carried out. The biological characterization showed that the scaffolds were compatible with MSCs and promoted osteoblast differentiation which was confirmed at both cellular and molecular levels. The cellular construct increased calcium deposition, analyzed by alizarin red staining and ALP activity at cellular level. At the molecular level, the osteoblast markers expression such as Runx2 and type 1 collagen mRNAs, and osteonectin (ON) and osteocalcin (OC) secretory proteins were increased in the presence of scaffold. Overall, the current study recommends that MSCs can be easily obtained from human waste OFF, and grown in standard in vitro conditions. Successful growth of such MSCs with CS/PCL/Zn scaffold opens new avenues in utilizing the cell source for bone tissue engineering.


Assuntos
Materiais Biocompatíveis , Regeneração Óssea/fisiologia , Líquido Folicular/fisiologia , Folículo Ovariano/fisiologia , Engenharia Tecidual/métodos , Tecidos Suporte , Adulto , Materiais Biocompatíveis/administração & dosagem , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/citologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/fisiologia , Células Cultivadas , Quitosana/administração & dosagem , Feminino , Líquido Folicular/citologia , Líquido Folicular/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais , Recuperação de Oócitos/métodos , Osteogênese/efeitos dos fármacos , Osteogênese/fisiologia , Folículo Ovariano/efeitos dos fármacos , Poliésteres/administração & dosagem , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Difração de Raios X/métodos , Zinco/administração & dosagem
2.
Parasitol Res ; 119(12): 4233-4241, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32996050

RESUMO

Albendazole is known as the drug of choice for medical treatment of cystic echinococcosis (CE). Albendazole sulfoxide (ABZ-SO), as the main active metabolite of albendazole, has low efficacy in the disease due to low water solubility and poor absorptivity. PLGA nanoparticles (NPs) enhance the dissolution of poorly soluble drugs, and chitosan (CS) coating enhances oral drug delivery of NPs. In this study, the efficacy of ABZ-SO-loaded CS-PGLA NPs in the treatment of CE was evaluated in laboratory mice. ABZ-SO-loaded CS-PGLA NPs were prepared by nanoprecipitation and characterized by dynamic light scattering method and scanning electron microscopy. Thirty mice were intraperitoneally infected by 1000 protoscoleces of Echinococcus granulosus. Ten months later, the mice were allocated into 3 groups: groups 1 and 2 were treated with ABZ-SO and ABZ-SO-loaded CS-PGLA NPs, respectively, and the mice in group 3 remained untreated as the control group. The drugs were administered by gavage for 45 days at a daily dose of 10 mg/kg. Finally, all mice were opened and the cysts were collected, counted, weighed, and measured separately. The therapeutic effect of ABZ-SO in the number, weight, and volume of the cysts were not statistically significant compared with those in ABZ-SO-loaded CS-PGLA NPs and the control group. However, the therapeutic effect of ABZ-SO-loaded CS-PGLA NPs in the weight and volume of cysts were statistically significant when compared with that in the control group (p ˂ 0.05). In conclusions, this study revealed that ABZ-SO-loaded CS-PGLA NPs could enhance the therapeutic efficacy of ABZ-SO in the treatment of CE in laboratory mice.


Assuntos
Albendazol/análogos & derivados , Antiplatelmínticos/administração & dosagem , Quitosana/química , Equinococose/tratamento farmacológico , Ácido Poliglicólico/química , Administração Oral , Albendazol/administração & dosagem , Albendazol/química , Animais , Antiplatelmínticos/química , Quitosana/administração & dosagem , Sistemas de Liberação de Medicamentos , Avaliação Pré-Clínica de Medicamentos , Echinococcus granulosus/efeitos dos fármacos , Camundongos , Nanopartículas/administração & dosagem , Nanopartículas/química , Ácido Poliglicólico/administração & dosagem
3.
PLoS One ; 15(9): e0238823, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32970684

RESUMO

Mucoadhesive polymeric nanocapsules have attracted interest of researchers from different fields from natural sciences because of their ability to interact with the mucosa and increase drug permeation. Anesthesia by immersion causes absorption through the skin and gills of fish, so it is important to evaluate the exposure of these organs to drug nanosystems. Benzocaine (BENZ) is one of the most popular anesthetic agents used in fish anesthesia, but it has drawbacks because of its low bioavailability, resulting in weak absorption after immersion. Here we describe method developed for preparing and characterizing chitosan-coated PLGA mucoadhesive nanoparticles containing BENZ (NPMAs) for zebrafish immersion anesthesia. We determined the lowest effective concentration, characterized the interaction of the mucoadhesive system with fish, measured the anesthetic efficacy, and evaluated possible toxic effects in embryos and adults exposed to the nanoformulations. This study opens perspectives for using nanoformulations prepared with BENZ in aquaculture, allowing reduction of dosage as well as promoting more effective anesthesia and improved interaction with the mucoadhesive system of fish.


Assuntos
Anestesia/veterinária , Benzocaína/administração & dosagem , Nanocápsulas/administração & dosagem , Peixe-Zebra , Animais , Aquicultura , Quitosana/administração & dosagem , Quitosana/toxicidade , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/toxicidade , Liberação Controlada de Fármacos , Brânquias/efeitos dos fármacos , Nanocápsulas/toxicidade , Pele/efeitos dos fármacos
4.
Anim Sci J ; 91(1): e13435, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32869472

RESUMO

This study aimed to investigate the effects of chitosan and whole raw soybean on nutrient intake, apparent digestibility, nitrogen utilization, microbial protein synthesis, blood metabolites, feeding behavior, ruminal fermentation, digesta kinetics, and reticular flow of nutrients of buffaloes. Four ruminally-cannulated Murrah buffaloes (351 ± 15 kg of initial BW) were randomly assigned according to a 4 × 4 Latin square design. Treatments were arranged as 2 × 2 factorial arrangement: the first factor was whole raw soybean (WRS), and the second factor was chitosan (CHI) with or without their inclusion in diets. Intake and apparent digestibility of ether extract (p < .01; p = .04, respectively), non-fiber carbohydrates intake (p = .03) and apparent ruminal digestibility of dry matter (p = .01) were affected by diets. An interaction effect or tendency was observed for microbial nitrogen (p = .09), concentrations, ruminal ammonia nitrogen (p = .05), total volatile fatty acid (p = .03). Association of chitosan with whole raw soybean has potential effects as a modulator of rumen fermentation; therefore, chitosan can be applied as an alternative non-ionophore for Murrah buffaloes.


Assuntos
Búfalos/fisiologia , Quitosana/administração & dosagem , Dieta/veterinária , Digestão , Fermentação , Nutrientes/metabolismo , Rúmen/metabolismo , Soja , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Ingestão de Alimentos , Masculino , Nitrogênio/metabolismo , Biossíntese de Proteínas
5.
Pharm Res ; 37(10): 195, 2020 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-32944793

RESUMO

PURPOSE: Design imiquimod-loaded chitosan nanocapsules for transdermal delivery and evaluate the depth of imiquimod transdermal absorption as well as the kinetics of this absorption using Raman Microscopy, an innovative strategy to evaluate transdermal absorption. This nanovehicle included Compritol 888ATO®, a novel excipient for formulating nanosystems whose administration through the skin has not been studied until now. METHODS: Nanocapsules were made by solvent displacement method and their physicochemical properties was measured by DLS and laser-Doppler. For transdermal experiments, newborn pig skin was used. The Raman spectra were obtained using a laser excitation source at 532 nm and a 20/50X oil immersion objective. RESULTS: The designed nanocapsules, presented nanometric size (180 nm), a polydispersity index <0.2 and a zeta potential +17. The controlled release effect of Compritol was observed, with the finding that half of the drug was released at 24 h in comparison with control (p < 0.05). It was verified through Raman microscopy that imiquimod transdermal penetration is dynamic, the nanocapsules take around 50 min to penetrate the stratum corneum and 24 h after transdermal administration, the drug was in the inner layers of the skin. CONCLUSIONS: This study demonstrated the utility of Raman Microscopy to evaluate the drugs transdermal penetration of in the different layers of the skin. Graphical Abstract New imiquimod nanocapsules: evaluation of their skin absorption by Raman Microscopy and effect of the compritol 888ATO® in the imiquimod release profile.


Assuntos
Quitosana/farmacocinética , Sistemas de Liberação de Medicamentos/métodos , Ácidos Graxos/farmacocinética , Imiquimode/farmacocinética , Nanocápsulas/administração & dosagem , Pele/metabolismo , Administração Cutânea , Animais , Quitosana/administração & dosagem , Quitosana/química , Ácidos Graxos/administração & dosagem , Ácidos Graxos/química , Imiquimode/administração & dosagem , Imiquimode/química , Nanocápsulas/química , Microscopia Óptica não Linear/métodos , Absorção Cutânea , Suínos
6.
AAPS PharmSciTech ; 21(7): 246, 2020 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-32856115

RESUMO

Enterococcus faecalis infections represent a health concern, mainly in oral diseases, in which treatments with chlorhexidine solution (0.2%) are often used; however, it presents high toxicity degree and several side effects. Based on this, the use of natural products as an alternative to treatment has been explored. Nonetheless, plant extracts have poor organoleptic characteristics that impair theirs in natura use. Therefore, this work aimed to evaluate the analytical profile, biological activity, and cytotoxicity in vitro of S. brasiliensis-loaded chitosan microparticles (CMSb) produced using different aspersion flow rates. The analytical fingerprint was obtained by FTIR and NIR spectra. Principal components analysis (PCA) was used to verify the similarity between the samples. The crystallinity degree was evaluated by X-ray diffraction (XRD). Phytochemical screening (PS) was performed to quantify phytocompounds. Antimicrobial activity was evaluated by minimum inhibitory concentration (MIC). Antibiofilm activity and bactericidal kinetics against E. faecalis (ATCC 29212 and MB 146-clinical isolated) were also assessed. The hemolytic potential was performed to evaluate the cytotoxicity. Data provided by FTIR, NIR, and PCA analyses revealed chemical similarity between all CMSb. Furthermore, the results from XRD analysis showed that the obtained CMSb present amorphous characteristic. Tannins and polyphenols were accurately quantified by the PS, but methodology limitations did not allow the flavonoid quantification. The low hemolytic potential assay indicates that all samples are safe. Antimicrobial assays revealed that CMSb were able to inhibit not only the E. faecalis ATCC growth but also the biofilm formation. Only one CMSb sample was able to inhibit the clinical strain. These results highlighted the CMSb antimicrobial potential and revealed this system as a promising product to treat infections caused by E. faecalis.


Assuntos
Anacardiaceae , Anti-Infecciosos/administração & dosagem , Quitosana/administração & dosagem , Enterococcus faecalis/efeitos dos fármacos , Microesferas , Extratos Vegetais/administração & dosagem , Administração Oral , Anti-Infecciosos/isolamento & purificação , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Enterococcus faecalis/fisiologia , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Humanos , Testes de Sensibilidade Microbiana/métodos , Tamanho da Partícula , Casca de Planta , Extratos Vegetais/isolamento & purificação
7.
AAPS PharmSciTech ; 21(6): 233, 2020 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-32794119

RESUMO

Endolysins are a novel class of antibacterials with proven efficacy in combating various bacterial infections, in vitro and in vivo. LysMR-5, an endolysin derived from phage MR-5, demonstrated high lytic activity in our laboratory against multidrug-resistant S. aureus (MRSA) and S. epidermidis strains. However, endolysin and proteins in general are associated with instability and short in vivo half-life, consequently limiting their usage as pharmaceutical preparation to treat bacterial infections. Nanoencapsulation of endolysins could help to achieve better therapeutic outcome, by protecting the proteins from degradation, providing sustained release, thus could increase their stability, shelf life, and therapeutic efficacy. Hence, in this study, the feasibility of alginate-chitosan nanoparticles (Alg-Chi NPs) to serve as drug delivery platform for LysMR-5 was evaluated. LysMR-5-loaded nanoparticles were prepared by calcium ion-induced pre-gelation of alginate core and its complexation with chitosan. The formation of nanoparticles was confirmed on the basis of DLS, zeta potential, and electron microscopy imaging. The LysMR-5-loaded nanoparticles presented a hydrodynamic diameter of 276.5 ± 42, a PDI of 0.342 ± 0.02, a zeta potential - 25 mV, and an entrapment efficiency of 62 ± 3.1%. The potential ionic interaction between alginate, chitosan, and LysMR-5 was investigated by FT-IR and SEM-EDX analysis. Using scanning electron microscopy (SEM) and transmission electron microscopy (TEM), nano-sized particles with characteristic morphology were seen. Different antibacterial assays and SDS-PAGE analysis showed no change in endolysin's structural integrity and bioactivity after entrapment. A direct antibacterial effect of blank Alg-Chi Nps, showing enhanced bactericidal activity upon LysMR-5 loading, was observed against S. aureus. At physiological pH (7.2), the release profile of LysMR-5 from Alg-Chi NPs showed a biphasic release and followed a non-Fickian release mechanism. The biocompatible nature as revealed by cytocompatibility and hemocompatibility studies endorsed their use as drug delivery system for in vivo studies. Collectively, these results demonstrate the potential of Alg-Chi NPs as nano-delivery vehicle for endolysin LysMR-5 and other therapeutic proteins for their use in various biomedical applications.


Assuntos
Alginatos/síntese química , Quitosana/síntese química , Nanopartículas/química , Infecções Estafilocócicas , Staphylococcus aureus/efeitos dos fármacos , Alginatos/administração & dosagem , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Quitosana/administração & dosagem , Relação Dose-Resposta a Droga , Sistemas de Liberação de Medicamentos/métodos , Endopeptidases/administração & dosagem , Endopeptidases/síntese química , Previsões , Humanos , Camundongos , Nanopartículas/administração & dosagem , Tamanho da Partícula , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/fisiologia
8.
PLoS One ; 15(8): e0237218, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32760143

RESUMO

Influenza is an infectious respiratory illness caused by influenza viruses. Despite yearly updates, the efficacy of influenza vaccines is significantly curtailed by the virus antigenic drift and antigenic shift. These constant changes to the influenza virus make-up also challenge the development of a universal flu vaccine, which requires conserved antigenic regions shared by influenza viruses of different subtypes. We propose that it is possible to bypass these challenges by the development of an influenza vaccine based on conserved proteins delivered in an adjuvanted nanoparticle system. In this study, we generated influenza nanoparticle constructs using trimethyl chitosan nanoparticles (TMC nPs) as the carrier of recombinant influenza hemagglutinin subunit 2 (HA2) and nucleoprotein (NP). The purified HA2 and NP recombinant proteins were encapsulated into TMC nPs to form HA2-TMC nPs and NP-TMC nPs, respectively. Primary human intranasal epithelium cells (HNEpCs) were used as an in vitro model to measure immunity responses. HA2-TMC nPs, NP-TMC nPs, and HA2-NP-TMC nPs (influenza nanoparticle constructs) showed no toxicity in HNEpCs. The loading efficiency of HA2 and NP into the TMC nPs was 97.9% and 98.5%, respectively. HA2-TMC nPs and NP-TMC nPs more efficiently delivered HA2 and NP proteins to HNEpCs than soluble HA2 and NP proteins alone. The induction of various cytokines and chemokines was more evident in influenza nanoparticle construct-treated HNEpCs than in soluble protein-treated HNEpCs. In addition, soluble factors secreted by influenza nanoparticle construct-treated HNEpCs significantly induced MoDCs maturation markers (CD80, CD83, CD86 and HLA-DR), as compared to soluble factors secreted by protein-treated HNEpCs. HNEpCs treated with the influenza nanoparticle constructs significantly reduced influenza virus replication in an in vitro challenge assay. The results indicate that TMC nPs can be used as influenza vaccine adjuvants and carriers capable of delivering HA2 and NP proteins to HNEpCs.


Assuntos
Adjuvantes Imunológicos/farmacologia , Quitosana/farmacologia , Vírus da Influenza A Subtipo H1N1/imunologia , Vacinas contra Influenza/farmacologia , Influenza Humana/prevenção & controle , Adjuvantes Imunológicos/administração & dosagem , Animais , Linhagem Celular , Células Cultivadas , Quitosana/administração & dosagem , Cães , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/farmacologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/administração & dosagem , Glicoproteínas de Hemaglutininação de Vírus da Influenza/farmacologia , Humanos , Vacinas contra Influenza/administração & dosagem , Influenza Humana/imunologia , Células Madin Darby de Rim Canino , Nanopartículas/administração & dosagem , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Proteínas de Ligação a RNA/administração & dosagem , Proteínas de Ligação a RNA/farmacologia , Proteínas do Core Viral/administração & dosagem , Proteínas do Core Viral/farmacologia
9.
Appl Environ Microbiol ; 86(18)2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32651210

RESUMO

The objective of this study was to evaluate the effect of chitosan microparticles on the uterine microbiome of cows with metritis. Dairy cows with metritis (n = 89) were assigned to 1 of 3 treatments: chitosan microparticles (n = 21), in which the cows received an intrauterine infusion of chitosan microparticles at metritis diagnosis (day 0), day 2, and day 4; ceftiofur (n = 25), in which the cows received a subcutaneous injection of ceftiofur on day 0 and day 3; and no intrauterine or subcutaneous treatment (n = 23). Nonmetritic cows (n = 20) were healthy cows matched with cows with metritis by the number of days postpartum at metritis diagnosis. Uterine swab samples collected on days 0, 3, 6, 9, and 12 were used for 16S rRNA gene sequencing and 16S RNA gene copy number quantification by quantitative PCR. Principal-coordinate analysis showed that the microbiome of the ceftiofur-treated and metritic untreated groups progressed toward that of the nonmetritic group by day 3, whereas that of the chitosan microparticle-treated group remained unchanged. The differences on day 3 were mainly due to a greater relative abundance of Fusobacteria, particularly Fusobacterium, in the chitosan microparticle-treated group than in the ceftiofur-treated and metritic untreated groups. Furthermore, the microbiome of the ceftiofur-treated group became similar to that of the nonmetritic group by day 9, whereas the microbiome of the chitosan microparticle-treated and metritic untreated groups became similar to that of the nonmetritic group only by day 12. The total bacterial 16S rRNA gene counts in the chitosan microparticle-treated group were greater than those in the metritic untreated controls on days 6 and 9, whereas the ceftiofur treatment group was the only group in which the total bacterial 16S rRNA gene count became similar to that in the nonmetritic group by day 12. In summary, chitosan microparticles slowed the progression of the uterine microbiome toward a healthy state, whereas ceftiofur hastened the progression toward a healthy state.IMPORTANCE Third-generation cephalosporins, such as ceftiofur, are commonly used to treat metritis in dairy cows. Chitosan microparticles has been shown to have a broad spectrum of activity in vitro and to be effective against uterine pathogens in vivo; therefore, they have been hailed as a possible alternative to traditional antibiotics. Nonetheless, in the present study, we saw that chitosan microparticle treatment slowed the progression of the uterine microbiome of cows with metritis toward a healthy state, whereas ceftiofur treatment hastened the progression toward a healthy state. Given the lack of an effective alternative to traditional antibiotics and an increased concern about antimicrobial resistance, a greater effort should be devoted to the prevention of metritis in dairy cows.


Assuntos
Doenças dos Bovinos/prevenção & controle , Quitosana/administração & dosagem , Endometrite/veterinária , Microbiota/efeitos dos fármacos , Nanopartículas/administração & dosagem , Útero/microbiologia , Animais , Bovinos , Endometrite/prevenção & controle , Feminino , Substâncias Protetoras/administração & dosagem
10.
Carbohydr Polym ; 239: 116236, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32414447

RESUMO

In the present work, hybrid microgels based on chitosan and SiO2 nanoparticles (NPs) were synthesized. Both chitosan and the SiO2 NPs were submitted to chemical modification reactions to having vinyl groups incorporated into their structures. The microgels were synthesized by emulsion polymerization. SEM analysis indicated a high dispersity of diameter for the microgels, ranging between (18.7 ±â€¯12.3) µm for the samples without SiO2-VTS and (11.3 ±â€¯8.07) µm for the microgels with SiO2-VTS. The material showed pH-responsiveness, especially in acidic pHs. The longest release lasted 45 min and large amounts of drugs were released as soon as the material was added to the release medium. It is interesting for oral drug delivery systems, especially for gastric wound treatment. The fast release of high amounts of drugs promotes an immediate relief of the pain and the following controlled release allows the gradual recovery of the damaged area.


Assuntos
Quitosana/química , Sistemas de Liberação de Medicamentos , Microgéis/química , Gastropatias/tratamento farmacológico , Vitamina B 12/farmacologia , Administração Oral , Sobrevivência Celular/efeitos dos fármacos , Quitosana/administração & dosagem , Células HT29 , Humanos , Concentração de Íons de Hidrogênio , Microgéis/administração & dosagem , Nanopartículas/administração & dosagem , Nanopartículas/química , Tamanho da Partícula , Silanos/administração & dosagem , Silanos/química , Dióxido de Silício/administração & dosagem , Dióxido de Silício/química , Propriedades de Superfície , Vitamina B 12/administração & dosagem , Vitamina B 12/química
11.
Poult Sci ; 99(1): 95-100, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32416857

RESUMO

This present experiment was performed to investigate the effects of dietary supplementation of chitosan (CS) on immune function in growing Huoyan geese. A total of 320 28-day-old healthy growing Huoyan geese (sex balance) with similar body weight were randomly allotted into control, CS100, CS200, and CS400 groups. Each group includes 4 replicates with 20 geese per replicate, and the feeding trial lasted for 4 wk. The 4 diets contained 0, 100, 200, and 400 mg CS per kg feed, respectively. The results showed that compared with the control group, the relative weight of thymus, serum concentrations of IGF-I, INS, GH, T3, T4, IgM, IgG, IgA, complement C3, and IL-2 in CS200 group were significantly higher at both 42 and 56 D of age, respectively (P < 0.05). In addition, relative weight of bursa of fabricius (BF), spleen, serum complement C4, and TNF-a concentrations in CS200 group were higher at 56 D of age (P < 0.05), no differences were observed at 42 D of age (P > 0.05). These results indicated that addition of 200 mg/kg CS enhanced immune organs weight, serum concentrations of immunoglobulins, complements, hormone, as well as cytokines, and improved immune function of growing Huoyan geese.


Assuntos
Proteínas Sanguíneas/metabolismo , Quitosana/metabolismo , Gansos/imunologia , Hormônios/sangue , Sistema Imunitário/efeitos dos fármacos , Ração Animal/análise , Animais , Quitosana/administração & dosagem , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Feminino , Gansos/crescimento & desenvolvimento , Gansos/metabolismo , Masculino , Distribuição Aleatória
12.
Biomater Sci ; 8(11): 3202-3211, 2020 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-32374304

RESUMO

Preventing surgical site infections (SSIs) of implants has drawn significant attention in both basic and clinical research. Implants with convenient preparation methods and intelligent drug release capabilities are highly needed to resist bacterial infection. Herein, we designed an intelligent drug-release system, which can be instantly incorporated with implants during the surgical process. The drug-release system involves ß-glycerophosphate (ß-GP) and chitosan (CS) as a thermosensitive hydrogel for instant construction onto implants and hyaluronic acid (HA) as a trigger to release vancomycin hydrochloride (VH) on demand. Tertiary calcium phosphate (TCP) scaffolds (implants) are vacuum-adsorbed in a solution of the intelligent vancomycin-release system (VH-HA-CS/ß-GP), followed by heating for 40 min at 80 °C to form VH-HA-CS/ß-GP@TCP. The drug-release hydrogel intelligently releases vancomycin depending on the concentration of hyaluronidase, which is secreted by Staphylococcus aureus (S. aureus) in infection sites. Furthermore, VH-HA-CS/ß-GP@TCP showed effective antibacterial properties in vitro and in vivo. The VH-HA-CS/ß-GP drug-release system can be conveniently prepared during surgery for intelligently preventing SSIs in bone tissue.


Assuntos
Antibacterianos/administração & dosagem , Doenças Ósseas/prevenção & controle , Sistemas de Liberação de Medicamentos , Implantes de Medicamento , Infecções Estafilocócicas/prevenção & controle , Infecção da Ferida Cirúrgica/prevenção & controle , Vancomicina/administração & dosagem , Animais , Antibacterianos/química , Osso e Ossos/cirurgia , Linhagem Celular , Quitosana/administração & dosagem , Quitosana/química , Liberação Controlada de Fármacos , Feminino , Glicerofosfatos/administração & dosagem , Glicerofosfatos/química , Ácido Hialurônico/administração & dosagem , Ácido Hialurônico/química , Hialuronoglucosaminidase/química , Hialuronoglucosaminidase/metabolismo , Hidrogéis/administração & dosagem , Hidrogéis/química , Masculino , Camundongos , Coelhos , Staphylococcus aureus/enzimologia , Vancomicina/química
13.
Sci Rep ; 10(1): 8360, 2020 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-32433572

RESUMO

Three different bioadhesive gels were evaluated in a double-blind randomized clinical trial in which microbial growth in the suture thread was assessed following post-surgical application of the aforementioned gels. Also assessed in this trial were, the intensity of post-surgical pain as well as the degree of healing of the patients' surgical wounds. A total of 21 patients (with 42 wisdom teeth) participated in this trial. Chlorhexidine gel, chlorhexidine-chitosan gel, and hyaluronic acid gel were evaluated, with a neutral water-based gel serving as the control agent. The aerobic and facultative anaerobic bacterial recovery on blood agar was lower in the placebo group than in the experimental groups. The most significant difference (p = 0.04) was observed in the chlorhexidine-chitosan group. in which the growth of Blood Agar and Mitis Salivarius Agar was significantly higher than in the placebo group. The intensity of post-surgical pain was very similar among all the groups. Significantly better healing rates were observed in the patients treated with chlorhexidine-chitosan gel when compared with those who used the placebo gel (p = 0.03), and in particular when compared with those patients who used hyaluronic acid gel (p = 0.01). Through our microbiological analyses, we were able to conclude that none of the bioadhesive gels tested resulted in beneficial reductions in the bacterial/fungal populations. However, the healing rates of patients who were treated with chlorhexidine-chitosan were better than those of the patients who used either the placebo gel or the hyaluronic acid gel.


Assuntos
Anti-Infecciosos Locais/administração & dosagem , Infecção da Ferida Cirúrgica/prevenção & controle , Suturas/microbiologia , Extração Dentária/efeitos adversos , Cicatrização/efeitos dos fármacos , Adolescente , Adulto , Quitosana/administração & dosagem , Clorexidina/administração & dosagem , Método Duplo-Cego , Feminino , Géis , Humanos , Ácido Hialurônico/administração & dosagem , Masculino , Medição da Dor , Dor Pós-Operatória/diagnóstico , Dor Pós-Operatória/etiologia , Dor Pós-Operatória/prevenção & controle , Placebos/administração & dosagem , Infecção da Ferida Cirúrgica/microbiologia , Adulto Jovem
14.
Carbohydr Polym ; 237: 116167, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32241399

RESUMO

Ternary composite films containing bulk chitosan (CS) and chitosan nanoparticles (CSNPs) with different concentrations were prepared using bacterial cellulose/poly(vinyl alcohol) as the base film and the composites films were compared. The micromorphology and mechanical, physical, chemical, antibacterial, and optical barrier properties of the composite films were compared. CS incorporation improved the mechanical properties, as the maximum tensile strength was increased to 130.55 ± 9.42 MPa. The dense structure of CSNPs prevented water diffusion and lessened the water content of the composite membranes. The inclusion of CS and CSNPs both reduced the water solubility and water vapor permeability. CS-doped films possessed good transparency, while CSNPs had better ultraviolet-barrier properties (3.84 % of transmittance at 200-280 nm). In addition, CSNPs-embedded membranes exhibited prominent antibacterial properties against Escherichia coli and Staphylococcus aureus, which were much greater than those of CS composite membranes with a maximum bacteriostatic diameter of 10.33 ± 1.55 mm.


Assuntos
Antibacterianos , Celulose , Quitosana , Nanopartículas , Antibacterianos/administração & dosagem , Antibacterianos/química , Celulose/administração & dosagem , Celulose/química , Quitosana/administração & dosagem , Quitosana/química , Escherichia coli/efeitos dos fármacos , Embalagem de Alimentos , Nanopartículas/administração & dosagem , Nanopartículas/química , Solubilidade , Staphylococcus aureus/efeitos dos fármacos , Resistência à Tração , Raios Ultravioleta
15.
Carbohydr Polym ; 237: 116162, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32241415

RESUMO

Electrical discharge plasma in a liquid phase can generate reactive species, e.g. hydroxyl radical, leading to rapid reactions including degradation of biopolymers. In this study, the effect of plasma treatment time on physical properties and cytotoxicity against cancer cells of N,O-carboxymethyl chitosan-stabilized gold nanoparticles (CMC-AuNPs) was investigated. AuNPs were synthesized by chemical reduction of HAuCl4 in 2 % CMC solution to obtain CMC-AuNPs, before being subjected to the plasma treatment. Results showed that the plasma treatment not only led to the reduction of hydrodynamic diameters of CMC-AuNPs from 400 nm to less than 100 nm by the plasma-induced degradation of CMC but also provided the narrow size distribution of AuNPs having diameters in the range of 2-50 nm, that were existing in CMC-AuNPs. In addition, the plasma-treated CMC-AuNPs could significantly reduce the percentage of cell viability of breast cancer cells by approximately 80 % compared to the original CMC and CMC-AuNPs.


Assuntos
Antineoplásicos/administração & dosagem , Quitosana/administração & dosagem , Ouro/administração & dosagem , Nanopartículas Metálicas/administração & dosagem , Antineoplásicos/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Quitosana/química , Eletroquímica , Ouro/química , Humanos , Nanopartículas Metálicas/química , Neoplasias/tratamento farmacológico
16.
Carbohydr Polym ; 237: 116170, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32241417

RESUMO

Poly (lactide-co-glycolide) (PLGA) nanoparticles surface functionalized with water soluble glycol chitosan (GC) and carboxymethyl chitosan (CMC) has been studied for their drug (Paclitaxel and Doxorubicin) loading, yield, cellular uptake, serum protein adsorption and hemocompatibility. It was observed that Paclitaxel (Ptxl) phase out as Extraneous Ptxl Precipitates (EPP) (>25 %) in case of uncoated and CMC coated low molecular weight (LMW) PLGA nanoparticles (PNPs). The EPP formation was significantly reduced to ∼5 % with GC coating as it enhanced LMW PLGA precipitation and yield predominantly spherical polymeric nanoparticles towards better encapsulation of Ptxl and thus uniform intracellular drug distribution. Interestingly, protein corona analysis showed cmcPNPs and gcPNPs to be distinct from each other in associating mainly with serum proteins of molecular weight < 30 kDa and >30 kDa respectively. While CMC functionalization showed >10 % hemolysis, at similar concentration GC coating was found to provide superior hemocompatibility even in the absence of protein corona.


Assuntos
Antineoplásicos Fitogênicos , Quitosana , Nanopartículas , Paclitaxel , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Precipitação Química , Quitosana/administração & dosagem , Quitosana/química , Liberação Controlada de Fármacos , Eritrócitos/efeitos dos fármacos , Cabras , Humanos , Nanopartículas/administração & dosagem , Nanopartículas/química , Paclitaxel/administração & dosagem , Paclitaxel/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/administração & dosagem , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Coroa de Proteína , Ratos
17.
Carbohydr Polym ; 237: 116163, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32241426

RESUMO

A polyelectrolyte complex nanoparticle comprising chitosan (CS) and carboxymethyl chitosan (CMCS) was prepared (CS/CMCS-NPs) by ionic gelation, which was then used as a doxycycline carrier (Dox:CS/CMCS-NPs). The obtained CS/CMCS-NPs and Dox:CS/CMCS-NPs were characterized for various parameters and bacteriostatic ability against Porphyromonas gingivalis. The regulation of related genes and proteins of NLRP3 inflammasome and IL-1ß in human gingival fibroblasts (HGFs) was characterized by qRT-PCR, western blotting and ELISA. The results showed that Dox:CS/CMCS-NPs had an orderly morphology and an excellent cytocompatibility. P. gingivalis was strongly inhibited by Dox:CS/CMCS-NPs contrasted with control group. Dox:CS/CMCS-NPs effectively down-regulated both gene and protein levels of NLRP3 inflammasome and IL-1ß in HGFs. This study provides a new method for rational application of Dox in the clinical treatment of periodontal disease and a new direction for explaining the mechanism of action of Dox:CS/CMCS-NPs and more drug-carrying nanoparticles.


Assuntos
Antibacterianos/administração & dosagem , Quitosana/análogos & derivados , Doxiciclina/administração & dosagem , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Nanopartículas/administração & dosagem , Doenças Periodontais/metabolismo , Adolescente , Antibacterianos/química , Células Cultivadas , Quitosana/administração & dosagem , Quitosana/química , Doxiciclina/química , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Inflamassomos/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Nanopartículas/química , Doenças Periodontais/genética , Porphyromonas gingivalis/efeitos dos fármacos , Porphyromonas gingivalis/crescimento & desenvolvimento
18.
Poult Sci ; 99(4): 2061-2067, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32241491

RESUMO

This experiment was conducted to investigate the effect of dietary chitosan oligosaccharide (COS) on growth performance, nutrient digestibility, jejunal morphology, gene expression, and plasma antioxidant enzymes in male broiler chickens under experimentally induced stress via in-feed dexamethasone (DEX). On day 3 after hatching, male broiler chicks were assigned to 2 diets supplemented with COS at 0 or 1 g/kg in a randomized complete block design and fed to day 27 after hatching. Birds were pooled within each diet (0 or 1 g/kg COS) to equalize the average BW and fed 2 diets supplemented with 0 or 1 g/kg DEX, within each dietary COS, from day 20 to 27 after hatching. This resulted in a 2 × 2 factorial arrangement of treatments with 2 levels each of COS and DEX, 8 replicate cages of 7 birds per cage. On day 27 after hatching, birds were weighed and euthanized, and samples were collected. Dietary COS decreased (P < 0.05) DEX-induced effects (interaction; P < 0.05) on BW, BW gain, and gain:feed. Dietary COS supplementation attenuated the DEX effects (interaction; P < 0.05) on villus height, crypt depth, villus height to crypt depth ratio, and ileal digestibility of dry matter and energy. The DEX-induced effect of relative mRNA expression of jejunal mucosa IL-6, IL-10, and claudin-1 was reduced by dietary COS supplementation (interaction; P < 0.05). Responses (interaction; P < 0.05) in the activity of plasma superoxide dismutase, catalase, and glutathione peroxidase to COS and DEX were similar to those observed with the relative mRNA expression. Chitosan oligosaccharide supplementation increased (P < 0.05) the mRNA expression of IL-8 and occludin. In conclusion, dietary COS decreased the DEX-induced effect by improving growth performance, nutrient digestibility, jejunal morphology, gene expression, and plasma antioxidant enzymes in broiler chickens. This implies that dietary COS may be useful for ameliorating the negative effect of stress on gut health in broiler chickens.


Assuntos
Antioxidantes , Galinhas/fisiologia , Quitosana/metabolismo , Imunidade/efeitos dos fármacos , Oligossacarídeos/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Ração Animal/análise , Animais , Quitosana/administração & dosagem , Dexametasona/farmacologia , Dieta/veterinária , Suplementos Nutricionais/análise , Imunossupressores/farmacologia , Masculino , Oligossacarídeos/administração & dosagem , Distribuição Aleatória , Estresse Fisiológico/fisiologia
19.
BMC Plant Biol ; 20(1): 113, 2020 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-32164536

RESUMO

BACKGROUND: Powdery mildew (PM) is an important disease of pea that reduce yield. Ascophyllum nodosum extract (ANE) and chitosan (CHT) are biostimulants used to improve plant health. Efficacy of ANE and CHT was assessed individually and in combination against pea powdery mildew. RESULTS: Combined applications of ANE and CHT had a significant inhibitory effect on pathogen development and it reduced disease severity to 35%, as compared to control (90.5%). The combination of ANE and CHT enhanced the activity of plant defense enzymes; phenylalanine ammonia lyases (PAL), peroxidase (PO) and production of reactive oxygen species (ROS) and hydrogen peroxide (H2O2). Further, the treatment increased the expression of a number of plant defense genes in jasmonic acid (JA) signaling pathway such as LOX1 and COI and salicylic acid (SA)-mediated signaling pathway such as NPR1 and PR1. Other genes involved in defense mechanisms like NADPH oxidase and C4H were also upregulated by the combination treatment. CONCLUSION: The combination of ANE and CHT suppresses pea powdery mildew largely by modulating JA and SA-mediated signaling pathways.


Assuntos
Ascomicetos/fisiologia , Ascophyllum/química , Quitosana/farmacologia , Ervilhas/imunologia , Doenças das Plantas/prevenção & controle , Imunidade Vegetal , Quitosana/administração & dosagem , Ervilhas/efeitos dos fármacos , Doenças das Plantas/microbiologia , Imunidade Vegetal/efeitos dos fármacos
20.
Vet Res ; 51(1): 37, 2020 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-32143695

RESUMO

Chitosan nanoparticles (CNPs) represent an efficient vaccination tool to deliver immunogenic antigens to the antigen-presenting cells (APCs), which subsequently stimulate protective immune responses against infectious diseases. Herein, we prepared CNPs encapsulating mRNA molecules followed by surface coating with conserved H9N2 HA2 and M2e influenza proteins. We demonstrated that CNPs efficiently delivered mRNA molecules into APCs and had effectively penetrated the mucosal barrier to reach to the immune initiation sites. To investigate the potential of CNPs delivering influenza antigens to stimulate protective immunity, we intranasally vaccinated chickens with empty CNPs, CNPs delivering HA2 and M2e in both mRNA and protein formats (CNPs + RNA + Pr) or CNPs delivering antigens in protein format only (CNPs + Pr). Our results demonstrated that chickens vaccinated with CNPs + RNA + Pr elicited significantly (p < 0.05) higher systemic IgG, mucosal IgA antibody responses and cellular immune responses compared to the CNPs + Pr vaccinated group. Consequently, upon challenge with either H7N9 or H9N2 avian influenza viruses (AIVs), efficient protection, in the context of viral load and lung pathology, was observed in chickens vaccinated with CNPs + RNA + Pr than CNPs + Pr vaccinated group. In conclusion, we show that HA2 and M2e antigens elicited a broad spectrum of protection against AIVs and incorporation of mRNAs in vaccine formulation is an effective strategy to induce superior immune responses.


Assuntos
Galinhas , Quitosana/administração & dosagem , Vírus da Influenza A Subtipo H9N2/imunologia , Vacinas contra Influenza/imunologia , Influenza Aviária/terapia , Doenças das Aves Domésticas/terapia , Administração Intranasal/veterinária , Animais , Nanopartículas/administração & dosagem , RNA Mensageiro/imunologia , RNA Viral/imunologia , Vacinação/veterinária
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