Micro and nanocrystalline cellulose based oral dispersible film; preparation and evaluation of in vitro/in vivo rapid release studies for donepezil

Oral fast-dispersible film was prepared by utlizing donepezil hydrochloride (drug) and various cellulose derivatives such as hydroxypropyl methyl cellulose (hypermellose) (HPMC), microcrystalline cellulose (MCC) and nanocrystalline cellulose (NCC) to treat Alzheimer's disease. NCC was synthesized by ultra-sonication method using MCC and this was converted to thinfilm formulation (NCC-F) using solvent casting technique. The interaction between the polymer and the drug was investigated by spectral analysis such as UV, FTIR, and 1H- NMR. FTIR confirmed that the compatibility of drug and polymer in ODF formulation. NCC-F has shown an average surface roughness of 77.04 nm from AFM and the average particle size of 300 nm from SEM analysis. Nano sized particle of NCC-F leads faster in vitro dissolution rate (94.53%) when compared with MCC-F and F3 formulation. Animal model (in vivo) studies of NCC-F formulation has reached peak plasma concentration (Cmax) up to 19.018 ng/mL in the span of (tmax) 4 h with greater relative bioavailability of 143.1%. These results suggested that high surface roughness with nanosized NCC-F formulation attained extended drug availability up to (t1/2) 70 h.

Preparation of chrysin solid lipid nanoparticles and their pharmacokinetic behaviors / 中成药

AIM To prepare chrysin solid lipid nanoparticles and to evaluate their pharmacokinetic behaviors.METHODS The particle size,Zeta potential and in vitro release rate of nanoparticles prepared by emulsification uhrasonication-low temperature solidification method were determined.Twelve SD rats were randomly divided into two groups and were intragastrically given suspensions of crude drug and nanoparticles,respectively.HPLC was used for the content determination of chrysin in plasma,after which blood drug concentration-time curves were drawn,and pharmacokinetic parameters were calculated.RESULTS The obtained nanoparticles demonstrated the particle size of (207.15 ±30.59) nm,PDI of (0.224 ±0.067) and Zeta potential of (-34.8 ±5.9) mV,respectively,whose accumulative release rate reached 84.36％ within 36 h.Their Cmax [(9.04 ± 1.52) μg/mL] and AUC0～t,[(33.67 ± 3.47) μg · h/mL] were much higher than those of the crude drug (P ＜ 0.01).CONCLUSION Solid lipid nanoparticles can promote the oral absorption and bioavailability of chrysin,with significant sustained-release characteristics.

Enhanced alkaline catalase production by Serratia marcescens FZSF01: enzyme purification, characterization, and recombinant expression

Background: Catalase (CAT) is an important enzyme that degrades H2O2 into H2O and O2. To obtain an efficient catalase, in this study, a new strain of high catalase-producing Serratia marcescens, named FZSF01, was screened and its catalase was purified and characterized. Results: After optimization of fermentation conditions, the yield of catalase produced by this strain was as high as 51,468 U/ml. This catalase was further purified using two steps: DEAE-fast flow and Sephedex-G150. The purified catalase showed a specific activity of 197,575 U/mg with a molecular mass of 58 kDa. This catalase exhibited high activity at 20­70°C and pH 5.0­11.0. Km of the catalase was approximately 68 mM, and Vmax was 1886.8 mol/min mg. This catalase was further identified by LC­MS/MS, and the encoding gene was cloned and expressed in Escherichia coli BL21 (DE3) with a production of 17,267 ± 2037 U/ml. Conclusions: To our knowledge, these results represent one of the highest fermentation levels reported among current catalase-producing strains. This FZSF01 catalase may be suitable for several industrial applications that comprise exposure to alkaline conditions and under a wide range of temperatures.

Preparation and Transdermal Absorption in vitro of Piroxicam Nanostructured Lipid Carrier / 中国药师

Objective: To prepare piroxicam nanostructured lipid carrier and investigate its transdermal absorption behavior in vitro. Methods:Piroxicam nanostructured lipid carrier was prepared by a melt-emulsion ultrasonication and low temperature-solidifica-tion method. The physicochemical properties such as appearance, morphology, particle size distribution, PdI and zeta potential of pi-roxicam nanostructured lipid carrier were evaluated. The transdermal absorption in vitro was investigated using Franz diffusion cells. Results:Piroxicam nanostructured lipid carrier was clear and transparent with small spherical shape as seen under a transmission elec-tron microscope. The particle size distribution, PdI and zeta potential was (106. 4 ± 31. 6) nm, (0. 217 ± 0. 07) and ( -31. 6 ± 2. 5) mV, respectively. Piroxicam nanostructured lipid carrier had higher cumulative transdermal amount in 12 h than piroxicam solution. Conclusion:The nanostructured lipid carrier can remarkably improve piroxicam permeation into skin, which provides reference for the new dosage form for the topical use of piroxicam.

Optimization of tilianin loaded solid lipid nanoparticles and its absorption and transport study in Caco-2 cell model / 中草药

Objective: To prepare and optimize tilianin loaded solid lipid nanoparticles (T-SLNs), and investigate the physicochemical properties, absorption and transport behavour of T-SLNs in vitro. Methods: T-SLNs were prepared by high shear homogenization followed by ultrasonication and optimized by central composite design and response surface methodology. In the study, the physicochemical properties of T-SLNs including size, polydispersity (PDI), Zeta potential, shape, entrapment efficiency and release profile in vitro were investigated, the absorption and transport behavour of T-SLNs in Caco-2 cell model were also measured. Results: The optimum formulation of T-SLNs consisted of: drug/lipid of 0.11, soy lecithin/lipid of 1.26, and content of tween-80 was 5.05%. The prepared T-SLNs were spherical and uniform with the mean particle diameter at (86.40 ± 0.62) nm, PDI (0.165 ± 0.080) and Zeta potential of (-24.2 ± 0.6) mV, respectively. The average EE was (89.81 ± 1.07)%, and the release in vitro showed that tilianin was released about (98.72 ± 1.57)% in 48 h. Besides, the absorption and transport assays of T-SLNs in Caco-2 cells model indicated that T-SLNs had a higher absorption and transport than tilianin. Conclusion: The method of high shear homogenization followed by ultrasonication is suitable for T-SLNs preparation. The optimal T-SLNs have a smaller particle size and high EE. Moreover, in the same concentration of tilianin, the absorption and transport amounts of T-SLNs in Caco-2 cell model were higher than tilianin.

Preparation of Celecoxib Nanostructured Lipid Carriers and Tissue Distribution in Rats / 中国药师

Objective:To prepare celecoxib nanostructured lipid carriers and investigate the characteristics of tissue distribution in rats. Methods:Celecoxib nanostructured lipid carriers were prepared by a melt-emulsion ultrasonication and low temperature-solidifi-cation method. The physicochemical properties of nanostructured lipid carriers were studied, such as particle size distribution, zeta po-tential and morphology. The concentration of celecoxib in different tissues was determined after tail vein injection of celecoxib nano-structured lipid carriers in rats. Results:The obtained celecoxib nanostructured lipid carriers were spherical with the average particle size of (103. 5 ± 32. 6) nm and zeta potential of ( -37. 3 ± 5. 1) mV. The re of celecoxib nanostructured lipid carriers in liver, spleen, brain and muscle respectively was 3. 43, 2. 99, 2. 38 and 2. 93 times higher than that of celecoxib injection. Conclusion:The biodistribution of celecoxib is changed by the nanostructured lipid carriers. Celecoxib nanostructured lipid carriers have the characteris-tics of liver, spleen and muscle targeting, which is benefit to improving the efficacy.

Comparative Study of Poly(L-Lactic Acid) Scaffolds Coated with Chitosan Nanoparticles Prepared via Ultrasonication and Ionic Gelation Techniques

In this study, an attempt was made to develop bi-functional constructs serving both as scaffolds and potential delivery systems for application in neural tissue engineering. The constructs were prepared in two steps. In the first step, the bulks of poly (L-lactic acid) (PLLA) in 1, 4-dioxane/water (87:13) were fabricated using liquid-liquid thermally induced phase separation technique. In the next step, the prepared bulks were coated with chitosan nanoparticles produced by two different techniques of ultrasonication and ionic gelation by grafting-coating technique. In ultrasonication technique, the chitosan solution (2 mg/mL) in acetic acid/sodium acetate buffer (90:10) was irradiated by an ultrasound generator at 20 kHz and power output of 750 W for 100 s. In ionic gelation technique, the tripolyphosphate in water solution (1 mg/mL) was added to the same chitosan solution. The physicochemical properties of the products were characterized by Scanning Electron Microscopy, Attenuated Total Reflection Fourier Transform-Infrared, liquid displacement technique, contact angle measurement, compressive and tensile tests, as well as zeta potential and particle size analysis using dynamic light scattering. Moreover, the cell proliferation and attachment on the scaffolds were evaluated through human glioblastoma cell line (U-87 MG) and human neuroblastoma cell line [BE (2)-C] culture respectively. The results showed that the samples coated with chitosan nanoparticles prepared by ultrasonication possessed enhanced hydrophilicity, biodegradation and cytocompatibility compared with pure PLLA and PLLA coated with chitosan nanoparticles prepared by ionic gelation. This study suggests successful nanoparticles-scaffold systems which can act simultaneously as potential delivery systems and tissue engineering scaffolds.

Preparation and in vitro Evaluation of Simvastatin-loaded Nanostructured Lipid Carriers / 中国药师

To prepare simvastatin nanostructured lipid carriers ( simvastatin-NLCs) . Methods:The simvastatin-NLCs were prepared by melt-emulsion ultrasonication and low temperature-solidification methods. Using the particle size, polydispersion in-dex, encapsulation efficiency and drug loading as the idices, the ratio of solid to liquid, lipid concentration, ratio of surfactant to cosur-factant, emulsifier concentration and drug concentration were optimized. The optimized simvastatin-NLCs were characterized for the en-capsulation efficiency, particle size, zeta potential and morphology. In vitro drug release behavior and stability of NLCs were also stud-ied. Results:The optimized simvastatin-NLCs formula was as follows:the concentration of simvastatin, cetyl palmitate, Miglyol? 812, soy lecithin and solutol HS15? was 0. 5%, 1. 5%, 4. 5%, 2. 5% and 1. 5%, respectively. The particle size and zeta potential of NLCs was (102. 2 ± 42. 1) nm and ( -33. 1 ± 4. 1) mV, respectively. The simvastatin-NLCs were found to be small and spherical with smooth surface under a transmission electron microscope. The in vitro release profile indicated that the accumulated release of sim-vastatin reached up to (59. 1 ± 4. 8) % in 24 h. The stability studies showed that simvastatin-NLCs were stable in 3 months after stored at 5℃. Conclusion:The formula of simvastatin-NLCs prepared by melt-emulsion ultrasonication and low temperature-solidifica-tion method is feasible.

The Change of Ginsenoside Composition in the Ginseng (Panax ginseng) Flower Buds by the Ultrasonication and Vinegar Process

The purpose of this study was to develop a new ginseng (Panax ginseng) flower buds extract with the high concentration of ginsenoside Rg3, Rg5, Rk1, Rh1 and F4, the Red ginseng special component. Chemical transformation from the ginseng saponin glycosides to the prosapogenin was analyzed by the HPLC. The ginseng flower buds were processed at the several treatment conditions of the ultrasonication (Oscillator 600W, Vibrator 600W) and vinegar (about 14% acidity). The result of UVGFB-480 was the butanol fraction of ginseng flower buds that had been processed with ultrasonication and vinegar for 480 minutes gained the highest amount of ginsenoside Rg5 (3.548%), Rh1 (2.037%), Rk1 (1.821%), Rg3 (1.580%) and F4 (1.535%). The ginsenoside Rg5 of UVGFB-480 was found to contain 14.3 times as high as ginseng flower buds extracts (GFB, 0.249%).

Optimization of Water Based-extraction Methods for the Preparation of Bioactive-rich Ginger Extract Using Response Surface Methodology.

Aims: Evaluation of three extraction methods to prepare bioactive-rich ginger extract for incorporation into a functional beverage. Study Design: Response surface methodology. Methodology: For the preparation of bioactive-rich ginger extract with water, conventional hot water extraction, ultrasonic-assisted extraction and high pressure homogenization-assisted extraction were evaluated. Response surface methodology was employed to optimize the extraction conditions of each method with respect to the highest polyphenols, antioxidant capacity (ferric reducing antioxidant power; FRAP) and percent inhibition of low density lipoprotein (LDL) cholesterol oxidation. Results: Multiple response optimizations revealed that the optimum extraction conditions for each extraction method were 60min extraction time under 55°C for hot water extraction, 15min ultrasonication under 52°C for ultrasonic-assisted extraction and 62°C under 140MPa homogenization pressure for high pressure homogenization-assisted extraction. Conclusion: The extract prepared from the ultrasonic-assisted extraction method exhibited the highest polyphenol recovery and antioxidant activity, compared to the extracts prepared from other two methods.

The Experimental Research in the Effects of Ultrasonication on Degradation of PLGA Released DNA / 中国生化药物杂志

Objective To identify the effective results of ultrasound in degradation of polymeric nanoparticles released DNA .Polymeric nanoparticles was made by dehydration of polyacetylglutamicacid (PLGA, polylactic-co-glycolic acid)solution. Method Green Fluorescent Protein (GFP) was enclosed by PLGA. Different kinds of ultrasound mode and different duct cycle and power ones were used to radiate PLGA solution for 90 s, 9 min, 20 min separately after the solution prepared for 2 hrs,then putted the solution on centrifugal machine at 13000 r/m. Using Choloroform to get rid of fat-soluble impurity,then applied nanodrop to survey the releasing rate of DNA. Finally the effect of cell expression were observed by fluorescent microscope. Results The amount of DNA released from PLGA in groups which were exposed to ultrasound were significantly different from the groups which were not exposed to ultrosound. The releasing amount of former groups had upper limitation. The releasing rate was increased with the increment of the irradiation time,frequency of ultrasound;The effect of the DNA releasing and PLGA degradation by continuous-wave irradiation was stronger than pulsed-wave ultrasound. Conclusion Ultrasound can promote the degradation of PLGA, and do help in DNA releasing and expression in vitro.

Statistical optimization of dithranol-loaded solid lipid nanoparticles using factorial design / Otimização estatística de nanopartículas lipídicas sólidas carregadas com ditranol utilizando planejamento fatorial

This study describes a 3² full factorial experimental design to optimize the formulation of dithranol (DTH) loaded solid lipid nanoparticles (SLN) by the pre-emulsion ultrasonication method. The variables drug: lipid ratio and sonication time were studied at three levels and arranged in a 3² factorial design to study the influence on the response variables particle size and percent entrapment efficiency ( percentEE). From the statistical analysis of data polynomial equations were generated. The particle size and percentEE for the 9 batches (R1 to R9) showed a wide variation of 219-348 nm and 51.33- 71.80 percent, respectively. The physical characteristics of DTH-loaded SLN were evaluated using a particle size analyzer, differential scanning calorimetry and X-ray diffraction. The results of the optimized formulation showed an average particle size of 219 nm and entrapment efficiency of 69.88 percent. Ex-vivo drug penetration using rat skin showed about a 2-fold increase in localization of DTH in skin as compared to the marketed preparation of DTH.

Este estudo descreve o planejamento factorial 3² para otimizar a formulação de nanopartículas lipídicas sólidas (SLN) carregadas com ditranol (DTH) pelo método da ultrassonificação pré-emulsão. As variáveis como proporção de fármaco:lipídio e o tempo de sonicação foram estudados em três níveis e arranjados em planejamento fatorial 3² para estudar a influência nas variáveis de resposta tamanho de partícula e eficiência percentual de retenção do fármaco ( por centoEE). Pela análise estatística, geraram-se equações polinomiais. O tamanho da partícula e a por centoEE para os 9 lotes (R1 a R9) mostraram ampla variação, respectivamente, 219-348 nm e 51,33-71,80 por cento. As características físicas das SLN carregadas com DTN foram avaliadas utilizando-se analisador de tamanho de partícula, calorimetria de varredura diferencial e difração de raios X. Os resultados da formulação otimizada mostraram tamanho médio de partícula de 219 nm e eficiência de retenção do fármaco de 69,88 por cento. A penetração ex vivo do fármaco utilizando pele de rato mostrou aumento de, aproximadamente, duas vezes na localização de DTH na pele, comparativamente à preparação de DTH comercializada.

Ultrasonication Versus High Pressure Homogenization in the Preparation of Ginkgolides A and B Long-circulating Solid Lipid Nanoparticles / 中国药房

OBJECTIVE:To study the preparative methods of the long-circulating solid lipid nanoparticles(LSLN)carrying ginkgolides A and B(GAB)and to study the physicochemical characteristics of the GAB-LSLN.METHODS:GAB-LSLN was prepared by ultrasonication or high pressure homogenization.Transmission electron microscopy was employed to study its shape.Particle size,zeta potential,and entrapment efficiency of GAB-LSLN were determined,and its stability after storage under room temperature for 4 weeks was determined as well.RESULTS:The GAB-LSLN prepared by ultrasonication was platelet-shaped and irregular,and that prepared by high pressure homogenization was spherical and regular in shape.The particle diameters of GAB-L SLN prepared by ultrasonication and high pressure homogenization were(219.6?14.3)nm and(173.9?10.4)nm respectively(P0.05).CONCLUSION:High pressure homogenization is superior to ultrasonication in that the prepared GAB-LSLN has small particle size,high stability and high entrapment efficiency.