Yi Jin

β-cyclodextrin-centered star-shaped amphiphilic polymers for doxorubicin delivery

AIMnDelivery of doxorubicin could be achieved by a novel micellar system based on beta-cyclodextrin-centered star-shaped amphiphilic polymers (sPEL/CD). This study specifically explored the effect of polylactide segments in sPEL/CD on various micelle properties, such as the critical micelle concentration, size, drug loading, cytotoxicity and drug resistance reversing effect.nnnMETHODnThe sPEL/CD was synthesized by the arm-first method. The critical micelle concentrations of polymeric micelles were determined by fluorescence spectrophotometry using pyrene as a probe. The oil/water method was applied to prepare doxorubicin-loaded micelles. 3-(4,5-dimethylthi-azol-2-yl)-2,5-diphenyltetrazolium bromide, confocal laser-scanning microscopy and flow cytometry were used to examine cell cytotoxicity and cellular uptake of the doxorubicin-loaded micelles. Finally, rhodamine-123 cellular uptake was determined to evaluate the polymer action on MCF-7 and MCF-7/ADR cells.nnnRESULTSnAll polymers exhibited low cytotoxicity and their micelles had a desirable release-acceleration pH (pH 5.0) for cytoplasmic drug delivery. With the introduction of polylactide into the polymer, the micelle critical micelle concentration can be effectively decreased and the drug-loading content was enhanced. Most importantly, the drug resistance of MCF-7/ADR cells was significantly reversed via the interaction between polymer and Pgp. Therefore, this type of polymer has potential superiority for cancer therapy.

Preparation and in vitro evaluation of liposomal chloroquine diphosphate loaded by a transmembrane pH-gradient method.

This study developed an active loading method for encapsulating chloroquine diphosphate (CQ) into liposomes. The effects of different formulation factors on the encapsulation efficiency (EE) and the size of CQ liposomes were investigated. These factors included the internal phase of liposomes, the external phase of liposomes, the ratio of drug to soybean phosphatidylcholine (drug/SPC), the ratio of cholesterol to soybean phosphatidylcholine (Chol/SPC), and the incubation temperature and time. The EE (93%) was obtained when using drug/SPC (1:50 mass ratio), SPC/Chol (1:5 mass ratio) at 0.10 M citrate-sodium citrate buffer (pH 3.6). As 5 mol% methoxypoly(ethylene glycol)(2,000) cholesteryl succinate (CHS-PEG(2000)) or distearoyl phosphatidylethanolamine-poly (ethylene glycol)(2,000) (DSPE-PEG(2000)) was added, the size of particle was reduced and the EE was improved. Freeze-drying with 5% trehalose as a cryoprotectant was carried out to achieve long-term stability. The drug release studies were performed in vitro simulating the desired application conditions, such as physiological fluids (pH 7.4), tumor tissues (pH 6.5) and endosomal compartments (pH 5.5). The release of CQ from the liposomes prepared via remote loading showed the significant pH-sensitivity and retention properties, which favored the application of liposomal CQ at tumor tissues and endosomal compartments.

Effect of Application Volume of Ethanol-Isopropyl Myristate Mixed Solvent System on Permeation of Zidovudine and Probenecid Through Rat Skin

Permeation of zidovudine (AZT) and probenecid from an ethanol-isopropyl myristate (IPM) mixed system through rat skin was studied in a finite system. Several volume sizes of the ethanol-IPM mixed systems containing AZT and probenecid, both as suspensions, were applied on the skin of the hairless rat using a vertical glass cell, and the fractions of the drugs permeated in 8 hr Q%,8hr were determined. For the systems containing 40% ethanol, the Q%,8hr value decreased with the reduction of volume of the system applied, and the decreasing profile was similar to that calculated on the assumption that the permeability of the drug does not change with the volume of the sample applied. On the other hand, in the systems containing 10% or 20% ethanol, the Q%,8hr value showed a maximum when a specific volume of the sample was applied. Therefore, the effect of sample volume on the Q%,8hr value was different between the 40% ethanol-IPM system and the 10% or 20% ethanol-IPM system. Following pretreatment of the skin with 0.105 ml/cm2 of drug-free 40% ethanol-IPM for 2 hr, several volume sizes of 10% ethanol-IPM systems containing the drugs were applied on the skin to explain why the different profiles were observed in the system containing 10% or 20% ethanol. The results for pretreated skin suggest that the amount of ethanol in the systems with low ethanol concentration and small application volume is too small to exert an effect that enhances permeation of the drugs. In those systems, the integrated effect of ethanol on the skin would be important for the enhancing effect. Total volume, as well as concentration, of an enhancer should be set precisely in designing an efficient transdermal delivery system.

Introduction of poly-L-lactic acid microspheres into the skin using supersonic flow: effects of helium gas pressure, particle size and microparticle dose on the amount introduced into hairless rat skin

A micro particulate bombardment system loaded with DNA‐ and RNA‐coated gold and tungsten micro particles (diameter 1–3 μm; density about 19 g cm−3), the Helios gene gun system (Helios gun system), has been used to deliver a gene into cells by accelerating the micro particles to high velocity using a supersonic flow of helium gas. To investigate whether drug‐loaded microspheres, > 20 μm in diameter and about 1.0 g cm−3 in density, could be delivered in powder form quantitatively into the skin using the Helios gun system equipped with a cartridge container fitted with a rupture membrane, we investigated the effect of the helium gas pressure in accelerating indometacin‐loaded poly‐L‐lactic acid (PLA) microspheres, as well as the particle size and the bombardment dose on delivery into the skin. Introduction of indometacin (i.e. indometacin‐loaded PLA microspheres) after bombardment, with 3.0 mg indometacin‐loaded PLA microspheres of a particle size of 20–38, 44–53 and 75–100 μM at a helium pressure of 100, 200 and 300 psi, of the abdomen of hairless rats increased in parallel with the helium pressure and it was also affected by the particle size, being highest at a diameter of 75–100 μm. However, introduction of higher amounts of PLA microspheres resulted in more severe skin erythema (skin damage) as monitored by the Draize score. Using lower bombardment doses (0.5 and 1.0 mg), the efficiency of introduction was improved and the skin damage markedly reduced. Moreover, discrete bombardment with a low dose provided a more efficient introduction of indometacin and less skin damage. These results suggest that bombardment injection of drug‐loaded microspheres in a powdered form by the Helios gun system appears to be a very useful tool for the quantitative delivery of a variety of drugs and an alternative to parenteral injection by needle, especially for delivering water‐soluble macromolecules.

Thermosensitive self-assembly behaviors of novel amphiphilic polyphosphazenes

A series of amphiphilic thermosensitive polyphosphazenes (PNIPAm-g-PPP) bearing N-isopropylacrylamide oligomers (oligo-PNIPAm) and glycine ethyl groups (GlyEt) as co-substituents have been synthesized via polymer substitute reaction. UV-visible spectra indicated that the aqueous solution of PNIPAm-g-PPP exhibited the lower critical solution temperature (LCST). Furthermore, the LCST was seriously influenced by the substitution ratios of PNIPAm to GlyEt in the copolymer. The more GlyEt the copolymer contained, the lower LCST it had. The critical association concentration (cac) of copolymers was determined by fluorescence probe method. It was found that cac was decreased with increasing GlyEt content of polyphosphazene. Also the formation of self-assembled micelles or nano-particles was confirmed by TEM.

An ovarian cell microcapsule system simulating follicle structure for providing endogenous female hormones.

The aim of this study was to create a microcapsule system simulating native follicle structure by introducing microcarrier culture to microencapsulation for providing endogenous female hormones. Granulosa and theca cells of rat follicles were isolated. Granulosa cells were grown on microcarriers and enclosed together with theca cells in alginate-chitosan-alginate microcapsules. The cell viability and female hormone secretion were investigated in vitro. The microcapsules were transplanted to ovariectomized rats and the serum levels of estradiol and progesterone were measured for 60 days. The microencapsulated granulosa cells growing on microcarriers exhibited enhanced viability and promoted secreting ability of estradiol and progesterone compared with those without the microcarriers. Co-microencapsulation of granulosa cells and theca cells markedly elevated estradiol secretion in vitro. Transplantation of co-microencapsulated granulosa cells on microcarriers and theca cells maintained serum estradiol and progesterone at normal levels for 60 days. Microcarrier cell culture has been proved to be an effective method to enhance the viability of granulosa cells in microcapsules. Moreover, the transplantation of microcapsules enclosing granulosa cells on microcarriers and theca cells may be promising to provide endogenous female hormones for menopausal syndrome treatment.

Endogenous release of female hormones from co-microencapsulated rat granulosa and theca cells

Deficiency of female hormones is the principal cause of menopausal syndrome. The aim of this study was to establish a co-microencapsulation model of ovarian granulosa cells (GCs) and theca cells (TCs), and to investigate its endogenous release of female hormones. Rat ovarian GCs and TCs were isolated and co-microencapsulated in alginate-chitosan-alginate microcapsules. The effects of cell number ratio of GCs/TCs on syntheses of estradiol (E2) and progesterone (P4) were investigated in vitro. Serum levels of E2 and P4 in ovariectomized rats were measured for 60xa0days after the co-microencapsulated GCs and TCs were transplanted. It was showed that E2 synthesis in vitro was influenced by cell number ratio of GCs/TCs and the ratio for the maximum synthesis was 1:2. Serum E2 and P4 levels could be maintained normal for 60xa0days by the co-microencapsulated GCs/TCs. Transplantation of co-microencapsulated GCs/TCs may be a promising approach to provide endogenous female hormones for menopausal syndrome.

Female hormone release of microencapsulated Xenopus laevis ovarian cells

Cell microencapsulation has potential for the treatment of endocrine diseases. This study aims to probe the feasibility of Xenopus laevis as an animal model for cell microencapsulation and transplantation and to evaluate the female hormone release of microencapsulated X. laevis ovarian cells. The cells were harvested, cultured and microencapsulated into alginate-chitosan-alginate microcapsules with an electrostatic generator. The estradiol and progesterone releases of the microencapsulated X. laevis ovarian cells were investigated both in vitro and in vivo. The results showed that the microencapsulated cells kept secreting estradiol and progesterone in vitro for 60 days. After transplantation, serum estradiol and progesterone levels in ovariectomized X. laevis remained elevated for 60 days. X. laevis has been proved to be a suitable animal model for cell microencapsulation and transplantation. Microencapsulated ovarian cells may be considered as a promising endogenous drug delivery system for the treatment of deficiency of female hormones.