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1.
Molecules ; 26(16)2021 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-34443336

RESUMO

We develop a suitable delivery system for niaouli essential oil (NEO) using a nanoemulsification method for acne vulgaris. Prepared nanoemulsions (NEs) were characterized for droplet dimension, rheology, surface charge, and stability. The ability of NEO formulations against Propionibacterium acnes and Staphylococcus epidermidis was investigated and all formulations showed antiacne potential in vitro. Ex vivo permeation studies indicated significant improvement in drug permeations and steady state flux of all NEO-NEs compared to the neat NEO (p < 0.05). On the basis of the studied pharmaceutical parameters, enhanced ex vivo skin permeation, and marked effect on acne pathogens, formulation NEO-NE4 was found to be the best (oil (NEO; 10% v/v); Kolliphor EL (9.25% v/v), Carbitol (27.75% v/v), and water (53% v/v)). Concisely, the in vitro and ex vivo results revealed that nanoemulsification improved the delivery as well as bioactivities of NEO significantly.


Assuntos
Portadores de Fármacos/química , Melaleuca/química , Nanoestruturas/química , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Emulsões , Óleos Voláteis/metabolismo , Permeabilidade , Pele/metabolismo , Staphylococcus epidermidis/efeitos dos fármacos
2.
Saudi Pharm J ; 28(10): 1253-1262, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33132719

RESUMO

Cell- based targeted delivery is recently gain attention as a promising platform for delivery of anticancer drug in selective and efficient manner. As a new biotechnology platform, bacterial ghosts (BGs) have novel biomedical application as targeted drug delivery system (TDDS). In the current work, Salmonellas' BGs was utilized for the first time as hepatocellular cancer (HCC) in-vitro targeted delivery system. Successful BGs loading and accurate analysis of doxorubicin (DOX) were necessary steps for testing the applicability of DOX loaded BGs in targeting the liver cancer cells. Loading capacity was maximized to reach 27.5 µg/mg (27.5% encapsulation efficiency), by incubation of 10 mg BGs with 1 mg DOX at pH 9 in constant temperature (25 °C) for 10 min. In-vitro release study of DOX loaded BGs showed a sustained release (182 h) obeying Higuchi sustained kinetic release model. The death rate (tested by MTT assay) of HepG2 reached to 64.5% by using of 4 µg/ml, while it was about 51% using the same concentration of the free DOX (P value < 0.0001 One-way ANOVA analysis). The proliferative inhibitory concentration (IC50) of the DOX combined formula was 1.328 µg/ml that was about one third of the IC50 of the free DOX (3.374 µg/ml). Apoptosis analysis (tested by flow-cytometry) showed more accumulation in early apoptosis (8.3%) and late apoptosis/necrosis (91%) by applying 1 µg/ml BGs combined DOX, while 1 µg/ml free DOX showed 33.4% of cells in early apoptosis and 39.3% in late apoptosis/necrosis, (P value˃ 0.05: one-way ANOVA). In conclusion, DOX loaded Salmonellas' BGs are successfully prepared and tested in vivo with promising potential as hepatocellular cancer (HCC) targeted delivery system.

3.
Drug Dev Ind Pharm ; 46(10): 1716-1725, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32893682

RESUMO

The use of bacterial ghosts (BGs) for drug delivery is an extremely fascinating perspective especially with the inherited efficient target-ability to specialized tissues. Trafficking of drug molecules across the outer membrane of Gram-negative bacteria are important to be understood for both loading (influx) and drug release (efflux). In this study, Escherichia coli (E. coli) BGs were prepared using modified protocol sponge-like reduced protocol (SLRP) which was used for loading of doxorubicin (DOX). First time in the literature, different possible factors affecting DOX loading from BGs were examined in this study. These factors including drug concentration, temperature, pH gradient, incubation time and tonicity, are proposed to effect on drug loading into E. coli BGs. Results of optimum effect from accompanied factors were found to be 10 mg/mL as DOX concentration at pH 6 with tonicity of 0.7% incubated overnight at 4 °C. After gather all factors, the amount of DOX loaded inside the BGs was recorded as 37.58%. The in vitro release studies of DOX loaded BGs over time showed a burst initial release rate of 26.75% at the first 12 h followed by a period of sustained release lasting for 16 days to give maximum release rate of 58.04%. Remarkably, DOX loaded in BG showed more apoptosis (55%) than control and DOX solution. Overall, the results indicated the presence of some important factors to be controlled when loading drugs into BGs. Also, data showed the future possibility of utilizing BGs to deliver DOX to colon cancer cells.


Assuntos
Portadores de Fármacos , Escherichia coli , Membrana Celular/metabolismo , Doxorrubicina/química , Doxorrubicina/metabolismo , Portadores de Fármacos/metabolismo , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Concentração de Íons de Hidrogênio
4.
3 Biotech ; 10(7): 325, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32656058

RESUMO

This study identifies the risk factors, microbiological properties, antimicrobial susceptibility patterns, mortality, and clinical complications associated with organisms causing diabetic foot infections (DFIs) with or without antibiotic treatment using data from a retrospective multicenter surveillance. Specimens collected from different hospitals were cultured and the extended-spectrum ß-lactamase (ESBL) excretion was estimated. The antibacterial susceptibility pattern and biofilm formation were completed along with the recommended standard methods. Overall, 792 diabetic foot patients (DFPs) were enrolled and a total of 1803 causative organisms were isolated. Polymicrobial infection was identified in 48.5% of the patients. The isolated Gram-positive pathogens (46.7%) were higher than Gram-negative (38.6%) or anaerobes (7.9%). The predominant pathogens were S. aureus (22.2%), methicillin-resistant S. aureus (7.7%), Enterococcus spp. (12.8%), Pseudomonas aeruginosa (9.4%), E. coli (7.9%), Klebsiella spp. (7.5%), Proteus mirabilis (8.9%), coagulase negative staphylococci (CoNS) (6.6%), anaerobic organisms (5.9%), and fungi (2.3%). Vancomycin and clindamycin exhibited no activity against Gram-positive bacteria. However, meropenem and imipenem displayed high activity against the Gram-negative isolates. Out of the 765 tested strains, 251 showed moderate (15.8%) to high (34%) level biofilm-producing phenotype. DFIs were widespread among the diabetic patients with different microbial etiology and the major organisms were aerobic organisms. Our findings may provide an insight into the development of appropriate therapeutic strategies for the management of DFIs.

5.
AAPS PharmSciTech ; 21(5): 168, 2020 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-32514657

RESUMO

Bacteriosomes are a member of cell-derived vesicles that are proposed as promising tools in diagnosis, therapy, and drug delivery. These vesicles could be derived from a virus, bacterial cells, and animal cells. Biotechnology techniques were used in bioengineering of cell-derived vesicles in vitro, and in vivo. Bacterial vesicles such as bacterial cells, bacterial ghost, or bacteriosomes are vesicular structures derived from bacteria produced by manipulation of bacterial cells by chemical agents or gene-mediated lysis. Subsequently, bacterial vesicles (bacteriosomes) are non-living, non-denatured bacterial cell envelopes free of the cytoplasm and genetic materials. Gram-negative and Gram-positive bacteria are exploited in the production of bacteriosomes. Bacteriosomes have instinct organs, tissues, cells, as well as subcellular tropism. Moreover, bacteriosomes might be used as immunotherapy and/or drug delivery shuttles. They could act as cargoes for the delivery of small drugs, large therapeutics, and nanoparticles to the specific location. Furthermore, bacteriosomes have nature endosomal escaping ability, hence they could traffic different bio-membranes by endocytosis mechanisms. Therefore, bacterial-derived vesicles could be used in therapy and development of an innovative drug delivery systems. Consequently, utilizing bacteriosomes as drug cargoes enhances the delivery and efficacy of administered therapeutic agents. This review highlighted bacteriosomes in terms of source, engineering, characterization, applications, and limitations.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Imunoterapia/métodos , Animais , Bactérias , Micropartículas Derivadas de Células , Humanos
6.
3 Biotech ; 10(2): 56, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32015952

RESUMO

India has got rich cultural inheritage in the forms of Ayurveda texts which are a rich and ample source of herbs, shrubs, trees and affluent in medicinally active phytoconstituents. Aconitum napellus is used for the cure of many ailments including rheumatoid arthritis, sciatica and gout. The present work attempts to evaluate the physicochemical and preliminary phytochemical studies on the tubers of Aconitum napellus along with its antidiabetic activity. The herbal standardization was carried out on the basis of organoleptic properties, physical characteristics and physicochemical properties. The body weight of ACON-I (1.25 mg/kg) and ACON-II (2.5 mg/kg) was recorded as 190.40 and 209.40 g, respectively, compared with 163.00 g in diabetic rats at day 28. The body weight of ACON-I and ACON-II was significantly increased compared with diabetic rats (p < 0.01). However, the body weight of ACON-I and ACON-II was decreased significantly (p < 0.01) compared with normal group (222.60 g). The blood glucose levels of diabetic rats and ACON-I group were recorded as 277.800 and 152.400 mg/dl, respectively, compared with 83.600 mg/dl in normal rats (p < 0.01). However, the HbA1c levels of diabetic rats and ACON-I group were recorded as 11.306 and 6.936% Hb, respectively, compared with 4.539% Hb in normal rats. The glucose and HbA1c levels of diabetic and ACON-I groups were significant compared with normal group (p < 0.01). The results of antidiabetic activity showed that the plant can be used as a potent source for the treatment of diabetes and its complications. The results of this work provided the referential information for the identification and standardization of Aconitum napellus along with its role as a hypoglycemic agent.

7.
Artif Cells Nanomed Biotechnol ; 47(1): 989-996, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30873877

RESUMO

Nano-erythrocyte coating has been developed as an interesting biomimetic platform to provide hybrid nano-carriers with innate functions to target liver cancer. This goal was achieved by coating nano-erythrocyte membranes (NEMs) onto 5-fluorouracil (5-FU)-loaded liposomes (LPs) to produce NEM-5-FU-LPs. This framework is used to promote the escape of 5-FU-LPs from degradation during systemic circulation. NEMs were obtained by hypotonic lysis of erythrocytes to produce ghost erythrocytes (GEs) followed by extrusion through polycarbonate membranes. Chimeric NEM-5-FU-LPs were fabricated via the fusion of NEMs and artificial LPs. The resultant chaperoned LPs were characterized based on particle size, morphology, entrapment efficiency (EE %), stability, protein content and phosphatidylserine exposure and their in vitro release profiles and cytotoxic efficacy were also determined. The present results revealed that 5-FU-LPs, NEM-5-FU and NEM-5-FU-LPs exhibited nanosize, spherical shapes and unimodal size distributions <0.3. In addition, the vesicles presented a zeta potential with EE% of 24.6-30.7% and an appropriate stability for 3 weeks. NEM-5-FU-LPs retained the erythrocyte membrane proteins as confirmed by PAGE and displayed a sustained release profile up to 48 h when compared to NEM-5-FU and the 5-FU solution. Moreover, hybrid NEM-5-FU-LPs induced a late cytotoxic effect after 48 h compared to the other formulations. Thus, mantling of 5-FU-LPs by NEMs could enhance vesicle controllability and their targetability to liver cancer cells.


Assuntos
Biomimética/métodos , Carcinoma Hepatocelular/patologia , Sistemas de Liberação de Medicamentos/métodos , Membrana Eritrocítica/química , Fluoruracila/química , Lipossomos/química , Neoplasias Hepáticas/patologia , Animais , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular , Preparações de Ação Retardada , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Fluoruracila/farmacocinética , Humanos , Lipossomos/farmacocinética , Neoplasias Hepáticas/metabolismo , Tamanho da Partícula , Ratos , Propriedades de Superfície
8.
AAPS PharmSciTech ; 20(2): 48, 2019 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-30617674

RESUMO

Bacterial ghosts (BGs) are non-deformed bacterial cell envelopes that possess undamaged external configurations for precise attachment to different cells of the human body. The Escherichia coli BGs were successfully produced using a modified sponge-like reduced protocol and characterized by SEM. Four different concentrations of 5-fluorouracil (5-FU) were used to study the impact on the "ghosts" cell wall. 5-FU was then loaded into the BGs and the loading capacity (LC %) and entrapment efficiency (EE %) were determined and were found to be 38.3 ± 0.8 and 76.6 ± 0.8, respectively. The in vitro release studies were conducted in dialysis bags over a time period of 16 days and the accumulative 5-FU released (%) was calculated. Overall, 69.2% of the ghost-associated 5-FU was released from the BGs and release from the E. coli ghosts is governed by non-Fickian diffusion. The Caco-2 cell line was used to investigate the cytotoxicity of 5-FU-loaded BGs.


Assuntos
Antimetabólitos Antineoplásicos/metabolismo , Neoplasias Colorretais/metabolismo , Portadores de Fármacos/metabolismo , Sistemas de Liberação de Medicamentos/métodos , Escherichia coli/metabolismo , Fluoruracila/metabolismo , Antimetabólitos Antineoplásicos/administração & dosagem , Antimetabólitos Antineoplásicos/química , Células CACO-2 , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Parede Celular/efeitos dos fármacos , Neoplasias Colorretais/tratamento farmacológico , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/química , Escherichia coli/química , Fluoruracila/administração & dosagem , Fluoruracila/química , Humanos , Testes de Sensibilidade Microbiana/métodos
9.
Saudi Pharm J ; 26(2): 232-237, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30166921

RESUMO

Bacterial ghosts (BGs) can be prepared by both genetic and chemical means. Genetic method include using lysis gene E. Chemical method include incubation with numerous agents for a short time at their minimum inhibitory or minimum growth concentrations (MIC or MGC). The aim of this study is to prepare the BGs with a new protocol via exposing the bacterial cells to tween 80 for an extended period of time followed by sudden reduction of the surrounding pH. Salmonella enterica serovar typhimurium ATCC 13311 was used for this purpose. The cells were incubated in 7% v/v tween 80 solution in Muller-Hinton broth for 24 h at 37 °C then pH was decreased to 3.6 by adding lactic acid for one hour. The bacterial pellets were separated by high speed centrifugation, and then washed three times by half normal saline solution. High quality BGs were visualized by scanning electron microscopy (SEM) revealing punctured cells with intact outer shells and at least one intramembranous tunnel. The absence of vital cells was confirmed by subculturing. The release of respective amounts of proteins and DNA is another evidence of ghost's production. In addition, the integrity of cells was proved by visualization of Gram-stained cells using light microscopy. In conclusion, this new protocol is simple, economic and feasible for BGs preparation.

10.
Talanta ; 154: 292-8, 2016 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-27154677

RESUMO

The objective of the present investigation was to develop and validate a 'green' reversed phase high-performance liquid chromatography (RP-HPLC) method for rapid analysis of a cytotoxic drug 5-fluorouracil (5-FU) in bulk drug, marketed injection, water-in-oil (w/o) nanoemulsion, double water-in-oil-in-water (w/o/w) nanoemulsion and bacterial ghost (BG) matrix. The chromatography study was carried out at room temperature (25±1°C) using an HPLC system with the help of ultraviolet (UV)-visible detector. The chromatographic performance was achieved with a Nucleodur 150mm×4.6mm RP C8 column filled with 5µm filler as a static phase. The mobile phase consisted of ethyl acetate: methanol (7:3% v/v) which was delivered at a flow rate of 1.0mLmin(-1) and the drug was detected in UV mode at 254nm. The developed method was validated in terms of linearity (r(2)=0.998), accuracy (98.19-102.09%), precision (% RSD=0.58-1.17), robustness (% RSD=0.12-0.53) and sensitivity with satisfactory results. The efficiency of the method was demonstrated by the assay of the drug in marketed injection, w/o nanoemulsion, w/o/w nanoemulsion and BG with satisfactory results. The successful resolution of the drug along with its degradation products clearly established the stability-indicating nature of the proposed method. Overall, these results suggested that the proposed analytical method could be effectively applied to the routine analysis of 5-FU in bulk drug, various pharmaceutical dosage forms and BG.


Assuntos
Antineoplásicos/análise , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Estabilidade de Medicamentos , Emulsões , Nanoestruturas , Reprodutibilidade dos Testes , Água
11.
Extremophiles ; 20(1): 79-90, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26515082

RESUMO

The strain Streptomyces sp. Al-Dhabi-1 was isolated from soil sediments collected from Tharban hot spring in the southern west of Saudi Arabia using actinomycetes isolation agar and starch casein agar at 55 °C. Identification of the isolate was done according to morphological, physiological and biochemical characteristics and 16S rRNA sequence similarity as well. 16S rRNA sequence and blast analyses confirmed that the isolate belonging to the genus Streptomyces. The sequence was submitted to GenBank with accession number (KF815080). Ethyl acetate extract of Streptomyces sp. Al-Dhabi-1 showed good antimicrobial activities against tested pathogenic microbes. Minimum inhibitory concentration results showed that the best values were observed against S. agalactiae (<0.039 mg/ml) and Klebsiella pneumonia (0.125 mg/ml). Minimum inhibitory concentration of Al-Dhabi-1 against fungi; Cryptococcus neoformans (0.078 mg/ml), C. albicans (0.156 mg/ml), A. niger (0.625 mg/ml), and T. mentagrophytes (0.156 mg/ml). GC-MS analysis was used for the chemical profile of ethyl acetate extract. Benzeneacetic acid (16.02 %) and acetic acid 2-phenylethyl ester (10.35 %) were the major compounds among 31 substances found the ethyl acetate extract. According to the results of antimicrobial activity against pathogenic microbes, it is clear that the actinomycetes from hot springs with extreme environments are promising source for antimicrobial compounds.


Assuntos
Antibiose , Fontes Termais/microbiologia , Microbiota , Streptomyces/isolamento & purificação , Sequência de Bases , Candida albicans/fisiologia , Cryptococcus neoformans/fisiologia , Genoma Bacteriano , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Streptomyces/genética , Streptomyces/metabolismo , Streptomyces/fisiologia
12.
Saudi Pharm J ; 22(3): 273-9, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25061413

RESUMO

Plackett-Burman randomization method is a conventional tool for variables randomization aiming at optimization. Bacterial Ghosts (BGs) preparation has been recently established using methods other than the E lysis gene. The protocol has been based mainly on using critical concentrations from chemical compounds able to convert viable cells to BGs. The Minimum Inhibition Concentration (MIC) and the Minimum Growth Concentration (MGC) were the main guide for the BGs preparation. In this study, Escherichia coli JM109 DEC has been used to produce the BGs following the original protocol. The study contained a detail protocol for BGs preparation that could be used as a guide.

13.
ScientificWorldJournal ; 2013: 545741, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23576904

RESUMO

Bacterial Ghosts (BGs) received an increasing interest in the recent years for their promising medicinal and pharmaceutical applications. In this study, for the first time we introduce a new protocol for BGs production. E. coli BL21 (DE3) pLysS (Promega) was used as a model to establish a general protocol for BGs preparation. The protocol is based on using active chemical compounds in concentrations less than the Minimum Inhibition Concentration (MIC). Those chemical compounds are SDS, NaOH, and H2O2. Plackett-Burman experimental design was used to map the best conditions for BGs production. Normal and electronic microscopes were used to evaluate the BGs quality (BGQ). Spectrophotometer was used to evaluate the amount of the released protein and DNA. Agarose gel electrophoresis was used to determine the existence of any residue of DNA after each BGs preparation. Viable cells, which existed after running this protocol, were subjected to lysis by inducing the lysozyme gene carried on pLysS plasmid. This protocol is able to produce BGs that can be used in different biotechnological applications.


Assuntos
Fracionamento Celular/métodos , Membrana Celular/efeitos dos fármacos , Membrana Celular/ultraestrutura , Escherichia coli/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Dodecilsulfato de Sódio/farmacologia , Hidróxido de Sódio/farmacologia , Escherichia coli/isolamento & purificação
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