Treatment of MPS I mice with microencapsulated cells overexpressing IDUA: effect of the prednisolone administration

Cell encapsulation, although a promising strategy to deliver therapeutic products, is hampered by immune response against biomaterials. The aim of this article is to assess the effect of prednisolone on enzyme release by microencapsulated cells implanted in vivo. Recombinant cells encapsulated were implanted in the peritoneum of wild-type mice and mucopolysaccharidosis (MPS) I mice, with or without prednisolone. Later, microcapsules were recovered for histological and enzyme analysis. Blood was collected from MPS I mice. All animals receiving prednisolone had a smaller inflammatory infiltrate. In vitro, prednisolone increased the amount of enzyme released from the recovered capsules, but this was not accompanied by an increase in the amount of circulating enzyme in vivo after 15 days. However, in 7 days, prednisolone significantly increased the amount of enzyme detected in the serum. Although prednisolone improved enzyme release in vitro and in vivo after 7 days, it was unable to maintain this effect for a longer period.     

Swelling behaviour and controlled drug release from cross-linked κ-carrageenan/NaCMC hydrogel by diffusion mechanism

We studied a model system of controlled drug release using beta-carotene and κ-carrageenan/NaCMC hydrogel as a drug and a device, respectively. Different concentrations of genipin were added to crosslink the beta-carotene loaded beads by using the dripping method. Results have shown that the cross-linked beads possess lower swelling ability in all pH conditions (pH 1.2 and 7.4), and swelling ratio decreases with increasing genipin concentration. Microstructure study shows that cross-linking has enhanced the stability and structure of the beads network. Determination of diffusion coefficient for the release of encapsulated beta-carotene indicates less diffusivity when beads are cross-linked. Swelling models using adaptive neuro fuzzy show that using genipin as a cross-linker in the kC/NaCMC hydrogels affects the transport mechanism. The model shows very good agreement with the experimental data that indicates that applying ANFIS modelling is an accurate, rapid and simple way to model in such a case for controlled release applications.     

Lyophilised Vegetal BM 297 ATO-Inulin lipid-based synbiotic microparticles containing Bifidobacterium longum LMG 13197: design and characterisation

Abstract

This study aimed at the manufacturing and characterisation of Vegetal BM 297 ATO-inulin-Bifidobacterium longum LMG 13197 microparticles prepared by freeze drying. Emulsions containing 1%, 1.5%, 2%, 3.5% or 5% w/v inulin were prepared, with or without centrifugation before freeze drying. Morphological properties, particle size distribution, encapsulation efficiency of the microparticles and their ability to preserve viability of the enclosed B. longum LMG 13197 cells were evaluated. The microparticles produced from both formulations without a centrifugation step were irregular, porous with concavities and contained high number of bacterial cells. Formulations with or without inulin had average particle sizes of 33.4–81.0?μm with encapsulation efficiencies of 82% and 88%, respectively. Vegetal-inulin microparticles have the morphology and size that will enable their even distribution in final food products, and hence, they have the potential for use as a functional food additive because they are likely to deliver sufficient numbers of viable bacteria.     

The effect of polymer molecular weight and cell seeding density on viability of cells entrapped within PEGDA hydrogel microspheres

Abstract

Cell microencapsulation can be used in tissue engineering as a scaffold or physical barrier that provides immunoisolation for donor cells. When used as a barrier, microencapsulation shields donor cells from the host immune system when implanted for cell therapies. Maximizing therapeutic product delivery per volume of microencapsulated cells necessitates first optimising the viability of entrapped cells. Although cell microencapsulation within alginate is well described, best practices for cell microencapsulation within polyethylene glycol is still being elucidated. In this study we microencapsulate mouse preosteoblast cells within polyethylene glycol diacrylate (PEGDA) hydrogel microspheres of varying molecular weight or seeding densities to assess cell viability in relation to cell density and polymer molecular weight. Diffusion studies revealed molecule size permissible by each molecular weight PEGDA towards correlating viability with polymer mesh size. Results demonstrated higher cell viability in higher molecular weight PEGDA microspheres and when cells were seeded at higher cell densities.     

Merits of sponge-like PLGA microspheres as long-acting injectables of hydrophobic drug

Abstract

Our study was initiated to challenge the preconception that nonporous PLGA microspheres with compact matrices should be used to develop long-acting depot injectables of hydrophobic drugs. A simple, new oil-in-water emulsion technique was utilized to produce porous PLGA microspheres with a sponge-like skeleton. Then, their applicability to developing sustained-release depots of hydrophobic drugs was explored in this study. As control, nonporous microspheres with a compact matrix were produced following a typical solvent evaporation process. Both microsphere manufacturing processes used non-halogenated isopropyl formate and progesterone as a dispersed solvent and a model hydrophobic drug, respectively. Various attempts were made to evaluate critical quality attributes of the porous microspheres and the nonporous ones. Surprisingly, the former displayed interesting features from the viewpoints of manufacturability and microsphere quality. For example, the spongy microspheres improved drug encapsulation efficiency and particle size uniformity, inhibited drug crystallization during microencapsulation, and minimized the residual solvent content in microspheres. Furthermore, the porous microspheres provided continual drug release kinetics without a lag time and much faster drug release than the non-porous microspheres did. In summary, the porous and sponge-like PLGA microspheres might find lucrative applications in developing sustained release dosage forms of hydrophobic drugs.     

Improving functional properties of pea protein isolate for microencapsulation of flaxseed oil

Unhydrolysed pea protein (UN) forms very viscous emulsions when used at higher concentrations. To overcome this, UN was hydrolysed using enzymes alcalase, flavourzyme, neutrase, alcalase–flavourzyme, and neutrase–flavourzyme at 50?°C for 0?min, 30?min, 60?min, and 120?min to form hydrolysed proteins A, F, N, AF, and NF, respectively. All hydrolysed proteins had lower apparent viscosity and higher solubility than UN. Foaming capacity of A was the highest, followed by NF, N, and AF. Hydrolysed proteins N60, A60, NF60, and AF60 were prepared by hydrolysing UN for 60?min and used further for microencapsulation. At 20% oil loading (on a total solid basis), the encapsulated powder N60 had the highest microencapsulation efficiency (ME?=?56.2). A decrease in ME occurred as oil loading increased to 40%. To improve the ME of N60, >90%, UN and maltodextrin were added. Flowability and particle size distribution of microencapsulated powders with >90% microencapsulation efficiency and morphology of all powders were investigated. This study identified a new way to improve pea protein functionality in emulsions, as well as a new application of hydrolysed pea protein as wall material for microencapsulation.     

Microencapsulation of anthocyanin-rich black soybean coat extract by spray drying using maltodextrin,gum Arabic and skimmed milk powder

Black soybean coat is insufficiently valorised food production waste rich in anthocyanins. The goal of the study was to examine physicochemical properties of spray dried extract of black soybean coat in regard to carrier materials: maltodextrin, gum Arabic, and skimmed milk powder. Maltodextrin and gum Arabic-based microparticles were spherical and non-porous while skimmed milk powder-based were irregularly shaped. Low water activity of microparticles (0.31–0.33), good powders characteristics, high solubility (80.3–94.3%) and encapsulation yields (63.7–77.0%) were determined. All microparticles exhibited significant antioxidant capacity (243–386 μmolTE/g), good colour stability after three months of storage and antimicrobial activity. High content of total anthocyanins, with cyanidin-3-glucoside as predominant, were achieved. In vitro release of anthocyanins from microparticles was sustained, particularly from gum Arabic-based. These findings suggest that proposed simple eco-friendly extraction and microencapsulation procedures could serve as valuable tools for valorisation and conversion of black soybean coat into highly functional and stable food colourant.     

Chitosan microencapsulation of the dispersed phase of an O/W nanoemulsion to hydrochlorothiazide delivery

In view of biopharmaceutical limitations of hydrochlorothiazide (HCTZ), Trojan-type mucoadhesive systems were proposed, aiming to improve HCTZ pharmacological properties by modulating its release. Nanoemulsions were formed spontaneously by combining medium-chain triglycerides (Lipoid^® S75 and Pluronic^® F68) and high encapsulation efficiency was obtained. The mucoadhesive properties were provided by chitosan and microencapsulation of nanoemulsions in spray-dryer was successfully achieved by using Aerosil^® as wall material. The rapid redispersion of nanoemulsion in simulated fluids led to a fast and complete release of HCTZ in gastric medium. The pharmacodynamics of HCTZ was improved, extending the diuretic activity. Once a simple and low-energy method contributed to obtain stable mucoadhesive nanoemulsions, advantages in terms of production could also be achieved, allowing easy scaling up. This novel mucoadhesive Trojan particulate system of HCTZ showed to be a promising approach to overcome limitations in terms of absorption and consequently improve the therapeutic efficacy.     

海藻酸钠-多聚赖氨酸-海藻酸钠微囊内细胞浓度与胰岛素和C肽的释放**

摘要 背景：微囊化已经普遍应用于各种实验研究,微囊包裹的干细胞治疗糖尿病问题也成为当今的热点,但是囊内包裹的细胞浓度与胰岛素释放量的关系成为目前需要解决的问题之一。 目的：运用海藻酸钠-多聚赖氨酸-海藻酸钠微囊包裹胰岛素产生细胞,观察细胞浓度对胰岛素和C肽释放情况的影响。 方法：制备大鼠胰腺损伤提取物,将小鼠骨髓间充质干细胞诱导分化为胰岛素产生细胞。免疫荧光和双硫腙染色鉴定诱导后细胞内胰岛素的表达。然后将胰岛素产生细胞制成浓度分别为1×107 L-1,5×107 L-1,1×108 L-1,5×108 L-1,1×109 L-1,5× 109 L-1的细胞悬液,气体吹喷法制成微囊,用6-羧基乙二酸荧光素检测囊内细胞活力,用葡萄糖刺激微囊内细胞检查其胰岛素和C肽的分泌情况。 结果与结论：胰腺损伤提取物诱导后双硫腙染色和细胞免疫荧光鉴定出胰岛素产生细胞内有胰岛素的表达;胰岛素产生细胞制成的微囊直径约为400 µm,大小均一,6-羧基乙二酸荧光素检测到囊内细胞的活力很好。用葡萄糖刺激不同细胞浓度的微囊,发现细胞浓度为1×108 L-1时,胰岛素和C肽的分泌达到最高。 关键词：微囊化;骨髓间充质干细胞;胰岛素产生细胞;胰岛素;C肽 doi:10.3969/j.issn.1673-8225.2011.03.046     

Development of small alginate microcapsules for recombinant gene product delivery to the rodent brain

A novel form of gene therapy using encapsulated recombinant cells in alginate microcapsules has proven effective in treating several animal models of human diseases. For treating neurological deficits in rodents with this technology, the size of the microcapsules has to be reduced for implantation in the central nervous system (CNS) to bypass the blood–brain barrier. This article reports the development of small alginate microcapsules suitable for implantation into the mouse CNS. By varying the encapsulation protocol, recombinant cells could be encapsulated in microcapsules ranging in diameter from 5 to 2000 μm. The optimal size for implantation was determined to be 100–200 μm, based on the smallest, homogeneously sized, cell-filled microcapsules that could pass the 500 μm inner diameter of a CNS-implantation needle. Compared with medium-sized (500–700 μm) microcapsules, these small microcapsules packed more tightly together with less inter-capsule space, resulting in an increased number of cells and a higher rate of recombinant gene product secretion per volume of microcapsules. The small microcapsules also displayed increased mechanical strength, compared with large microcapsules. These excellent in vitro properties of small 100–200 μm microcapsules warrant further in vivo investigation into the feasibility of using immuno-isolation gene therapy to deliver recombinant gene products to the rodent CNS.