Optimization of Clostridium tyrobutyricum encapsulation by extrusion method and characterization of the formulation

Purpose: To optimize the process parameters for the encapsulation of Clostridium tyrobutyricum (Ct) and to determine its in vitro characteristics.Methods: The process parameters, including the concentration of the wall and hardening material, Ct to gelatin ratio and hardening time, were studied by single factor analysis, while optimization was performed by orthogonal experimental design for the encapsulation rate of Ct.Results: Optimal conditions exhibited by orthogonal experimental design at a 92.17 % encapsulation rate with a viable count of 9.61 ± 0.06 lgCFU/g were: 6 % modified starch, 3 % sodium alginate, and 2 % CaCl2 at a Ct to gelatin ratio of 1:1 with a hardening time of 30 min. The survival rates of encapsulated Ct were higher than free Ct in simulated gastric (6.22 %) and intestinal juices (15.55 %). Reduction in viable counts of Ct at 90 °C were higher for free cells (44.76 %) than encapsulated cells (28.09 %) after 30 min of heat treatment. Correspondingly, encapsulation boosted the capacity of Ct to withstand the strong acidic conditions of the stomach and improved the storage properties of Ct.Conclusion: The results suggested that extrusion is a good technique for the encapsulation of Ct, as it enhances the viability of Ct during their transit through the gastrointestinal tract. Furthermore, encapsulation is favorable for Ct if planned for use in formulations where high temperature treatment is required.

Loading Effect of Chitosan Derivative Nanoparticles on Different Antigens and Their Immunomodulatory Activity on Dendritic Cells

Drug carrier nanoparticles (NPs) were prepared by the polyelectrolyte method, with chitosan sulfate, with different substituents and quaternary ammonium chitosan, including C236-HACC NPs, C36-HACC NPs, and C6-HACC NPs. To evaluate whether the NPs are suitable for loading different antigens, we chose bovine serum albumin (BSA), ovalbumin (OVA), and myoglobin (Mb) as model antigens to investigate the encapsulation effect of the NPs. The characteristics (size, potential, and encapsulation efficiency) of the NPs were measured. Moreover, the NPs with higher encapsulation efficiency were selected for the immunological activity research. The results showed that chitosan derivative NPs with different substitution sites had different loading effects on the three antigens, and the encapsulation rate of BSA and OVA was significantly better than that of Mb. Moreover, the NPs encapsulated with different antigens have different immune stimulating abilities to DCS cells, the immune effect of OVA-coated NPs was significantly better than that of BSA-coated NPs and blank NPs, especially C236-HACC-OVA NPs. Furthermore, we found that C236-HACC-OVA NPs could increase the phosphorylation level of intracellular proteins to activate cell pathways. Therefore, C236-HACC NPs are more suitable for the loading of antigens similar to the OVA structure.

Preparation and Sustained-Release Performance of PLGA Microcapsule Carrier System

Microcapsules have been widely studied owing to their biocompatibility and potential for application in various areas, particularly drug delivery. However, the size of microcapsules is difficult to control, and the size distribution is very broad via various encapsulation techniques. Therefore, it is necessary to obtain microcapsules with uniform and tailored size for the construction of controlled-release drug carriers. In this study, emulsification and solvent evaporation methods were used to prepare a variety of ovalbumin-loaded poly (lactic-co-glycolic acid) (PLGA) microcapsules to determine the optimal preparation conditions. The particle size of the PLGA microcapsules prepared using the optimum conditions was approximately 200 nm, which showed good dispersibility with an ovalbumin encapsulation rate of more than 60%. In addition, porous microcapsules with different pore sizes were prepared by adding a varying amount of porogen bovine serum albumin (BSA) to the internal water phase. The release curve showed that the rate of protein release from the microcapsules could be controlled by adjusting the pore size. These findings demonstrated that we could tailor the morphology and structure of microcapsules by regulating the preparation conditions, thus controlling the encapsulation efficiency and the release performance of the microcapsule carrier system. We envision that this controlled-release novel microcapsule carrier system shows great potential for biomedical applications.

Codelivery of Emodin and Diammonium Glycyrrhizinate by Anti-alpha8 Integrin-Conjugated Immunoliposomes for the Treatment of Renal Fibrosis

The targeted delivery of therapeutics to the kidneys has a profound potential for the management of renal fibrosis. Thus, we developed a drug delivery system that targets mesangial cells by conjugating anti-alpha8 integrin to the surface of liposomes. We coloaded emodin (EMO) and diammonium glycyrrhizinate (DAG) to the immunoliposomes for combined therapy. The coloaded immunoliposomes were small size (92.4±0.4 nm), narrowly distributed, and with nearly neutral zeta potential and good stability. The encapsulation rate of EMO and DAG in immunoliposomes was 45.5±2.0% and 44.3±1.1%, respectively. Using a BCA assay, the actual number of antibody molecules attached to a single liposome was determined as being approximately 41. An in vitro release study showed that EMO and DAG could be ratiometrically released from the immunoliposomes, which means that an optimized synergistic ratio of the two drugs could be achieved. Studies on cellular uptake studies demonstrated an approximately 3-fold increase for immunoliposomes in HBZY-1 cells compared to nonconjugated liposomes. In vitro cell growth inhibition and Western Blot assay revealed that the coloaded immunoliposomes exhibited a stronger and synergistic in vitro antifibrosis effect against NIH3T3 and HBZY-1 cells in vitro. Taken together, it indicated that anti-alpha8 integrin-modified immunoliposomes for codelivery of EMO and DAG have great potential for targeting the kidneys for the treatment of renal fibrosis.

Simvastatin-loaded nanoemulsions: development, characterization, stability study and toxicity assays

Aim: The aim of this study is to prepare and characterize simvastatin-loaded nanoemulsions (SIM-LN) as well as evaluate their physicochemical properties and toxicity. Methodology & results: The SIM-LN were prepared, their characteristics evaluated for 30 days, and after that, the SIM-LN toxicity was evaluated using Vero cell culture and the in vivo model of Caenorhabditis elegans. The prepared SIM-LN had an average droplet size of 139 ± 22 nm, with high encapsulation rate (>98.4%). The storage at room temperature proved to be the most optimal condition. Toxicity assays demonstrated no toxicity. Conclusion: It was demonstrated that the surfactants used as emulsifiers optimized the properties without side effects, because no toxicity was measured in preliminary tests.

Development and In Vitro Evaluation of Linear PEI-Shelled Heparin/Berberine Nanoparticles in Human Osteosarcoma U-2 OS Cells

Berberine (BBR), a natural isoquinoline alkaloid derived from Chinese herbs, exerts many biological effects, including antiviral, antimicrobial, antidiarrhea, anti-inflammatory, and antitumor effects. In this study, a novel berberine nanoparticle (NP) consisting of heparin (HP) and BBR with or without being shelled with linear polyethyleneimine (LPEI) was developed to enhance its antitumor activity on osteosarcoma U-2 OS cells. With varying ratios of HP to BBR, HP/BBR NPs had a size ranging from 218.4 ± 3.9 to 282.0 ± 5.1 nm and zeta potential from −35.7 ± 0.4 to −51.9 ± 1.8 mV. After shelling with LPEI, the resultant NPs (HP/BBR/LPEI) possessed a size ranging from 226.3 ± 3.0 to 405.7 ± 85.2 nm and zeta potential from −46.5 ± 0.3 to −35.6 ± 0.5 mV; the encapsulation rate of BBR was close to 80%. The release profiles of both NPs were revealed to be slower than that of BBR solution. Results also showed that BBR and its two derived NPs reduced the viability of U-2 OS cells, and BBR NPs increased the cellular uptake of BBR. Cells were arrested at the G1 phase when treated individually with BBR and the two NPs (HP/BBR and HP/BBR/LPEI) and DNA condensation was induced. In addition, BBR and BBR NPs reduced the expression of mouse double minute 2 homolog (MDM2) but increased that of p53, and BBR NPs enhanced apoptotic effects. In short, heparin-based nanoparticles could be potential carriers for osteosarcoma treatment.

Development and In Vitro Evaluation of Linear PEI-Shelled Heparin/Berberine Nanoparticles in Human Osteosarcoma U-2 OS Cells

Berberine (BBR), a natural isoquinoline alkaloid derived from Chinese herbs, exerts many biological effects, including antiviral, antimicrobial, antidiarrhea, anti-inflammatory, and antitumor effects. In this study, a novel berberine nanoparticle (NP) consisting of heparin (HP) and BBR with or without being shelled with linear polyethyleneimine (LPEI) was developed to enhance its antitumor activity on osteosarcoma U-2 OS cells. With varying ratios of HP to BBR, HP/BBR NPs had a size ranging from 218.4 ± 3.9 to 282.0 ± 5.1 nm and zeta potential from −35.7 ± 0.4 to −51.9 ± 1.8 mV. After shelling with LPEI, the resultant NPs (HP/BBR/LPEI) possessed a size ranging from 226.3 ± 3.0 to 405.7 ± 85.2 nm and zeta potential from −46.5 ± 0.3 to −35.6 ± 0.5 mV; the encapsulation rate of BBR was close to 80%. The release profiles of both NPs were revealed to be slower than that of BBR solution. Results also showed that BBR and its two derived NPs reduced the viability of U-2 OS cells, and BBR NPs increased the cellular uptake of BBR. Cells were arrested at the G1 phase when treated individually with BBR and the two NPs (HP/BBR and HP/BBR/LPEI) and DNA condensation was induced. In addition, BBR and BBR NPs reduced the expression of mouse double minute 2 homolog (MDM2) but increased that of p53, and BBR NPs enhanced apoptotic effects. In short, heparin-based nanoparticles could be potential carriers for osteosarcoma treatment.

Novel facile self-assembly approach to construct graphene oxide-decorated phase-change microcapsules with enhanced photo-to-thermal conversion performance

Novel self-assembled phase change microcapsule exhibits high encapsulation rate, large heat storage capacity, good thermal reliability and enhanced photo-thermal performance.

Effect of nutritional stress and sex on melanotic encapsulation rate in the sexually size dimorphic Cook Strait giant weta (Deinacrida rugosa)

Nutritional condition and sex are known to influence efficacy and investment in immune function. A poor diet is costly to immune function because it limits the resources (e.g., protein) available to effector systems (e.g., melanotic encapsulation), whereas males and females are expected to differ in how they allocate resources to fitness-related traits. Males are expected to invest less in immunity, and more in mating, than females, but this pattern could be reversed if fitness is more condition-dependent in males than in females. I tested the effects of nutritional condition and sex on melanotic encapsulation rate in the Cook Strait giant weta (Deinacrida rugosa Buller, 1871), an orthopteran insect exhibiting strong female-biased sexual size dimorphism that is, at least in part, the result of strong sexual selection for small male size. I found that male D. rugosa have a stronger encapsulation response than females, while nutritional condition has only a small positive effect on this particular effector system in both sexes. Whether the observed sex difference in encapsulation ability is due to a physiological constraint in females or whether males allocate more resources to this effector system because their fitness is more condition-dependent than female’s remains to be determined.

Antheraea pernyi Silk Fibroin Microspheres Carried Lysozyme

A method was developed to prepare the Antheraea Pernyi silk fibroin (ASF) microspheres using lysozyme as a model drug to estimate the application of ASF in drug controlled release field. The structure of ASF microsphere carried drug was characterized by X-ray diffraction and FTIR. The morphology and the influence of the microspheres on the degradation of lysozyme were investigated using scanning electron microscope. The results show that encapsulation rate was increased and drug content was decreased with the addition of lysozyme increasing. In vitro release of lysozyme from the ASF particles we demonstrated that the release kinetics depends on the pH. The pH played important roles in controlling lysozyme release profiles. It also can be seen that the degradation speed of lysozyme ASF microspheres slightly larger than the pure ASF microspheres.