In vitro evaluation and in vivo performance of lyophilized gliclazide

Enhancement of the dissolution rate of the poorly water-soluble hypoglycemic agent, gliclazide, by the aid of lyophilization was investigated. Mannitol, sodium lauryl sulfate (SLS) and polyvinyl pyrrolidone (PVP-k-30) were employed in different weight ratios (43%, 56% and 64% w/w, respectively) as water-soluble excipients in the formulation. Lyophilized systems were found to exhibit extremely higher in vitro dissolution rate compared to the unprocessed drug powder. Solid state characterization of the lyophilized systems using X-ray powder diffraction, Fourier transform infrared spectroscopy and differential scanning calorimetry techniques revealed that dissolution enhancement was attributable to transformation of gliclazide from the crystalline to an amorphous state in the solid dispersion formed during the lyophilization process. The gastrointestinal absorption and hypoglycemic effect of the lyophilized gliclazide/SLS system were investigated following oral administration to Albino rabbits. C_max and area under the plasma concentration–time curve of gliclazide (AUC_0–12) after administration of the lyophilized formulations were significantly higher than those obtained after administration of the unprocessed gliclazide.     

Dynamic modeling of the secondary drying stage of freeze drying reveals distinct desorption kinetics for bound water

In freeze drying, the desorption step for reaching a low target moisture content may take a significant fraction of the total process duration. Because the long-term stability of freeze-dried biological products strongly depends on the current moisture content, modeling the desorption process may help safely optimize the secondary drying step. Most published models assume a first-order desorption kinetic, but experimental evidence shows that strongly bound water in the monolayer takes a much longer time to be desorbed than less bound water in multilayer. The proposed model for desorption of freeze-dried lactic acid bacteria preparation accounts for monolayer and multilayer water state in the solid matrix, with very different desorption kinetics. Results showed that the ratio of characteristic desorption times (monolayer/multilayer) was almost 30. Temperature dependence was adequately described by an Arrhenius law in the range of 15 to 40°C. Model parameter identification used simultaneously gravimetric measurements with high time resolution and direct Karl-Fisher titration, from several experiments at different, time-varying temperatures.     

Lyophilized flutamide dispersions with polyols and amino acids: preparation and in vitro evaluation

Context: Flutamide (FLT) has poor aqueous solubility and low oral bioavailability. Objective: Lyophilization monophase solution was used for preparing lyophilized dispersions of FLT with polyols and amino acids to increase its poor dissolution. Methods: Physical properties and dissolution behavior of their physical mixtures and lyophilized dispersions were investigated. Results and discussion: The carriers increased the aqueous solubility of FLT but to a limited extent with arginine and glycine showing a linear A_L-phase solubility diagrams. Gas chromatography indicated that residual tertiary butyl alcohol was in range of 0.007?0.023% (w/w) in the dispersions. In all dispersions, the crystal structure of FLT was confirmed using differential scanning calorimetry, X-ray diffractometry, and scanning electron microscopy. However, the percent drug crystallinity was found to decrease with increasing the carrier content. Infrared spectroscopy revealed no interaction between drug and carriers. The particle size of FLT dispersions ranged between 0.61 and 1.81 μm, with a high surface area (293.93?465.37 m²/g) and porosity (447.69?754.33 e^-3 mL/g). In addition, the poor flow properties of FLT were improved but to a limited extent. FLT dissolution from the dispersions was enhanced with 46.35% and 36.43% of FLT dissolved after 30 minutes from 1:5 FLT–mannitol and FLT–trehalose dispersions, respectively, compared with only 13.45% of pure FLT. On the other hand, after 30 minutes 38.57% and 46.78% of FLT was dissolved from 1:3 FLT–arginine and FLT–glycine dispersions, respectively. Conclusion: These data suggest that polyols and amino acids might be useful adjuncts in preparation of immediate-release formulations of FLT.     

Preparation and evaluation of lyophilized liposome-encapsulated bufadienolides

Objective: The objective of this study was to prepare bufadienolides-loaded liposome (BU-lipo). Methods: The BU-lipo was prepared by a thin-film hydration method involving sonication and lyophilization procedures. The lyophilized BU-lipo was characterized with regard to the appearance and particle size by scanning electron microscopy, transmission electron microscopy, and photon correlation spectroscopy. The entrapment efficiency (EE) of BU-lipo was evaluated by the microdialysis technique. Results: In the optimal formulation, Lipoid E-80® and the mass ratio of cholesterol to lipid were fixed at 1.25% and 0.05. The media diameters of BU-lipo before and after lyophilization were about 100 nm, and the EEs of bufalin (B), cinobufagin (C), and resibufogenin (R) were 86.5%, 90.0%, and 92.1%, respectively. In the EE study, the probe recoveries of B, C, and R were 21.53?±?1.14%, 19.49?±?1.34%, and 20.19?±?1.25%, respectively, at a flow rate of 4 μL/min by the gain method. The EE of BU-lipo evaluated by microdialysis and ultrafiltration were equivalent. Conclusion: The lyophilized BU-lipo contained trehalose (10%) was stable up to 6 months in a desiccator under 2ºC–8ºC. The microdialysis technique has a wide application perspective in the investigation of the free-drug concentration of microcarrier systems.     

Ultrasonication, Lyophilization, Freezing and Storage Effects on Fat Loss during Mechanical Infusion of Expressed Human Milk

Ultrasonic homogenization was extended to situations where expressed human milk needs to be stored before being administered. We investigated whether the effect of ultrasonication would persist during storage in the frozen or lyophilized form. Recovery of fat was higher in ultrasonicated and frozen milk (stored for both 1 and 4 mo), than in milk stored following ultrasonication and lyophilization. The low fat recovery from stored lyophilized milk was increased by ultrasonicating the milk after storage and reconstitution (instead of prior to storage). Protein recovery was virtually complete with both methods.     

A Review of: “Superheated Steam Drying” by Mikhailov Yu.A. Energia,Moscow, 1967, 199 p. (in Russian)

This paper presents a general overview of Freeze-Drying (Lyophilization) and discusses the underlying principles of the process. Emphasis has been placed on the freeze-drying of biological materials and special mention has been made to the formulation of bio products prior to processing     

蛋白质药品冷冻干燥技术研究进展

冷冻干燥技术是制备固体蛋白质药品的常用方法。本文综述了蛋白质药品冻干保护机理、冻干工艺、冻干机的研究进展，并提出了优化药品冷冻干燥过程，提高药品的稳定性和经济性的研究思路。     

Application of lyophilization to prepare the nitrifying bacterial biofilm for imaging with scanning electron microscopy

Structure of bacterial biofilms may be investigated using several variants of scanning electron microscopy (SEM). We apply lyophilization to prepare nitrifying bacterial biofilm for conventional SEM imaging in high-vacuum mode (CSEM). We therefore replace standard biofilm fixation in glutaraldehyde cross-linking, ethanol dehydration, and critical-point drying (CPD) with less-invasive low-temperature drying by sublimation in vacuum. We compare this approach with: (1) standard preparation with glutaraldehyde fixation, ethanol dehydration, and CPD before CSEM, (2) cryo-sputter preparation of rapidly frozen biofilm in hydrated state (cryo-SEM), and (3) in situ observation without any sample pretreatment in environmental SEM. Combined imaging with these modalities revealed two distinct immobilization patterns on the polyurethane foam: (1) large irregular aggregates (flocs) of bacterial biofilm that exist as irregular biofilm fragments, rope-like structures, or biofilm layers on the foam surface; (2) biofilm threads adherent to the surface of polyurethane foam. Our results indicate that lyophilization was suitable for preservation of bacterial cells and many forms of structure of extracellular matrix. The lyophilized material could be imaged with high resolution (using CSEM) to generate structural information complementary to that obtained with other SEM techniques.     

Silk Biomaterials with Vascularization Capacity

Functional vascularization is critical for the clinical regeneration of complex tissues such as kidney, liver, or bone. The immobilization or delivery of growth factors has been explored to improve vascularization capacity of tissue‐engineered constructs; however, the use of growth factors has inherent problems such as the loss of signaling capability and the risk of complications including immunological responses and cancer. Here, a new method of preparing water‐insoluble silk protein scaffolds with vascularization capacity using an all‐aqueous process is reported. Acid is added temporally to tune the self‐assembly of silk in the lyophilization process, resulting in water‐insoluble scaffold formation directly. These biomaterials are mainly noncrystalline, offering improved cell proliferation than previously reported silk materials. These systems also have an appropriate softer mechanical property that could provide physical cues to promote cell differentiation into endothelial cells, and enhance neovascularization and tissue ingrowth in vivo without the addition of growth factors. Therefore, silk‐based degradable scaffolds represent an exciting biomaterial option, with vascularization capacity for soft tissue engineering and regenerative medicine.     

响应面法优化植物乳杆菌冻干保护剂

采用响应面法对植物乳杆菌冻干保护剂配比进行优化。在单因素试验的基础上,首先利用Plackett -Burman设计法研究保护剂对响应值的影响程度,发现脱脂乳、甘油和VC 对细胞存活率的影响显著,然后利用最陡爬坡法逼近最大响应区域,最后在上升最高点处由中心组合试验和响应面分析确定其最优保护剂复配比例,即脱脂乳、甘油和VC 的质量浓度分别为13.68g/100mL、1.97g/100mL 和0.20g/100mL,并通过实验测得优化后的细胞存活率为85.01%,与预测值84.85% 非常接近,植物乳杆菌含菌量为4.98 × 1010 CFU/g,而加单一保护剂时的细胞存活率最高值为52%,存活率提高了63.48%。