Toshiaki Dobashi

Thermosensitive polymer-conjugated albumin nanospheres as thermal targeting anti-cancer drug carrier

Thermosensitive Poly(N-isopropylacrylamide-co-acrylamide-co-allylamine) (PNIPAM-AAm-AA)-conjugated albumin nanospheres (PAN) was developed as a new thermal targeting anti-cancer drug carrier by conjugating PNIPAM-AAm-AA on the surface of albumin nanospheres (AN). AN with diameter below 200nm and narrow size distribution was successfully prepared in the first step with desolvation technique. PNIPAM-AAm-AA with different molecular weight (M(w)) was synthesized in the second step by radical polymerization and conjugated onto the surface of AN. Anti-cancer drug adriamycin (ADR) was then entrapped into the AN and PAN during the particle preparation. Compared with AN, the release rate of ADR from PAN in trypsin solution was slower, and decreased with increasing the conjugation amounts (hairy density) or M(w) of PNIPAM-AAm-AA (hairy length). Moreover, the release of ADR from PAN above the cloud-point temperature (T(cp)) of PNIPAM-AAm-AA became faster due to shrinkage of hairy thermosensitive polymer. To testify the thermal targetability in vivo, PAN was incubated with HepG2 cells. As expected, PAN can target cancer cells above the T(cp) of PNIPAM-AAm-AA, whereas it cannot below the T(cp). These results might reflect that PAN may selectively accumulate onto solid tumors that are maintained above physiological temperature due to local hyperthermia.

A galactosamine-mediated drug delivery carrier for targeted liver cancer therapy

In order to minimize the side effect of cancer chemotherapy, a novel galactosamine-mediated drug delivery carrier, galactosamine-conjugated albumin nanoparticles (GAL-AN), was developed for targeted liver cancer therapy. The albumin nanoparticles (AN) and doxorubicin-loaded AN (DOX-AN) were prepared by the desolvation of albumin in the presence of glutaraldehyde crosslinker. Morphological study indicated the spherical structure of these synthesized particles with an average diameter of around 200 nm. The functional ligand of galactosamine (GAL) was introduced onto the surfaces of AN and DOX-AN via carbodiimide chemistry to obtain GAL-AN and GAL-DOX-AN. Cellular uptake and kinetic studies showed that GAL-AN is able to be selectively incorporated into the HepG2 cells rather than AoSMC cells due to the existence of asialoglycoprotein receptors on HepG2 cell surface. The cytotoxicity, measured by MTT test, indicated that AN and GAL-AN are non-toxic and GAL-DOX-AN is more effective in HepG2 cell killing than that of DOX-AN. As such, our results implied that GAL-AN and GAL-DOX-AN have specific interaction with HepG2 cells via the recognition of GAL and asialoglycoprotein receptor, which renders GAL-AN a promising anticancer drug delivery carrier for liver cancer therapy.

Pinning of phase separation of aqueous solution of hydroxypropylmethylcellulose by gelation

Abstract Opalescence of the aqueous solution of hydroxypropylmethylcellulose (HPMC) induced by heating has been studied in terms of the phase diagram and the phase separation dynamics. The cloud point curve and the sol-to-gel transition curve intersected with each other at about 55 °C . Just above the cloud-point curve at which the spinodal curve has its minimum, a ring-like scattering pattern appeared corresponding to the spinodal decomposition. Temporal growth of the scattering function in the course of phase separation was studied by a time-resolved light scattering technique. The gelation pinned the phase separation (spinodal decomposition) of the aqueous HPMC solution.

Reagent-free crosslinking of aqueous gelatin: manufacture and characteristics of gelatin gels irradiated with gamma-ray and electron beam.

In order to obtain a gelatin hydrogel crosslinked by a reagent-free method, gamma-ray and electron beam radiation was applied to porcine, bovine and fish gelatin gels and the products were characterized by measuring the gel fraction, the swelling ratio and the enzymatic degradability. On increasing the radiation dose, the gel fraction increased and both the swelling ratio and the enzymatic degradability decreased. The transition temperature from gel to sol of the hydrogel containing more than 5% mammal gelatins increased up to more than 90°C when gamma-ray or electron beam were irradiated by more than 10 kGy. The results show that the degree of crosslinking of irradiated gelatin hydrogels increases with increasing irradiation dose and with decreasing concentration. It is suggested that the radiation crosslinking occurs around the physical crosslinking point or multiple helix structure of gelatin gel.

Anisotropic structure of calcium-induced alginate gels by optical and small-angle X-ray scattering measurements

It was more than 50 years ago that an appearance of birefringence in alginate gels prepared under cation flow was reported for the first time, however, the anisotropic structure of the alginate gel has not been studied in detail. In the present study, anisotropic Ca-alginate gels were prepared within dialysis tubing in a high Ca(2+)-concentration external bath, and optical and small-angle X-ray scattering (SAXS) measurements were performed to characterize the structure of the gel. The observations of the gel with crossed polarizers and with circular polarizers revealed the molecular orientation perpendicular to the direction of Ca(2+) flow. Analyses of the SAXS intensity profiles indicated the formation of rod-like fibrils consisting of a few tens of alginate molecules and that the anisotropy of the gel was caused by the circumferential orientation of the large fibrils. From the observed asymmetric SAXS pattern, it was found that the axis of rotational symmetry of the anisotropic structure was parallel to the direction of Ca(2+) flow. The alignment factor (A(f)) calculated from the SAXS intensity data confirmed that the orientation of the fibrils was perpendicular to the direction of Ca(2+) flow.

Release characteristics of an azo dye from poly(ureaurethane) microcapsules.

The time course for the transfer of azo dyes from the microcapsule core of dioctylphthalate to the dispersing medium of methanol through a poly(ureaurethane) (PUU) membrane was measured for various average size microcapsules with a size distribution. The dye release curves are represented by a stretched exponential function C(t)=C(eq)[1-exp [-(t/tau(eff))(alpha)]], where C(t) is the concentration of the dye in methanol, C(eq) that at the equilibrium state, and t the time. The exponent alpha decreased by increasing the variance of the size distribution of microcapsules. The effective time constant tau(eff) was expressed by tau(eff)=c(0)/kappa(alpha), where and kappa(alpha) are the mean square of microcapsule radius and a correction for the size distribution, respectively. The characteristic proportional constant, c(0), was determined as 2.6+/-0.2 min/microm(2). From the value of the constant, the diffusion coefficient of the azo dye in the PUU membrane was evaluated as 2.6 x 10(-12) cm(2)/s.

An approach to analysis of pigment release from microcapsules with size distribution

We have studied the release curve for microcapsules with size distribution. On the basis of an analogy to the relaxation phenomena with multiple characteristic times, we propose a stretched exponential release curve for the system with size distribution and relate the release curve to the size distribution function of the microcapsule. This method was successfully applied to the transfer of azo-pigments from inner medium of dioctyl phthalate to dispersing medium of methanol through poly(ureaurethane) microcapsule membrane.

Studies on the Formation Mechanism and the Structure of the Anisotropic Collagen Gel Prepared by Dialysis-Induced Anisotropic Gelation

We have found that dialysis of 5 mg/mL collagen solution into the phosphate solution with a pH of 7.1 and an ionic strength of 151 mM [corrected] at 25 °C results in a collagen gel with a birefringence and tubular pores aligned parallel to the growth direction of the gel. The time course of averaged diameter of tubular pores during the anisotropic gelation was expressed by a power law with an exponent of 1/3, suggesting that the formation of tubular pores is attributed to a spinodal decomposition-like phase separation. Small angle light scattering patterns and high resolution confocal laser scanning microscope images of the anisotropic collagen gel suggested that the collagen fibrils are aligned perpendicular to the growth direction of the gel. The positional dependence of the order parameter of the collagen fibrils showed that the anisotropic collagen gel has an orientation gradient.

Coexistence curve of polystyrene in methylcyclohexane. VII. Coexistence surface and critical double point of binary system in T–p–φ space

Cloud‐point measurements were made for binary systems of polystyrene (PS) +methylcyclohexane (MCH) with three PS samples in the pressure range up to 90 MPa. On a temperature (T)–pressure (p) plane, observed cloud‐point curves, which were determined at various compositions for the molecular weight Mw=1.61×104 and at the critical composition for Mw=1.02×104 and 3.49×104, decrease first, reach a minimum and increase with increasing pressure. For Mw=1.61×104 the coexistence surface was investigated in a T–p–φ space, where φ denotes the volume fraction of PS. On a T–φ plane the coexistence curves give a critical composition independent of pressure. On a p–φ plane one phase region appears between two coexistence curves. The top and the bottom of the coexistence curves give an upper critical solution pressure (UCSP) and a lower critical solution pressure (LCSP), respectively. The two critical points coincide forming a critical double point (CDP) at T=19.28 °C, p=48.6 MPa, and φ=0.1719. The shape of the coexisten...

Culture scale-up studies as seen from the viewpoint of oxygen supply and dissolved carbon dioxide stripping.

Oxygen supply and dissolved carbon dioxide (dCO(2)) stripping are two of the most important control parameters in cell culture. In this study, we investigated the effect of scale-up on the volumetric gas transfer coefficient with bioreactors of different sizes (working volume: 80 L, 500 L, 2000 L, and 10,000 L; aspect ratio: 1.0-1.6). Sparging air into water increased the volumetric oxygen transfer coefficient (k(L)a), an index of oxygen supply efficiency, by scale-up roughly in proportion to the depth of the water. A corresponding increase in k(L)a was found in a real cell culture of Chinese hamster ovary cells. dCO(2) stripping efficiency was evaluated in water tests using changes in k(L)a(co2), an index defined in relation to k(L)a. k(L)a(co2) increased following surface aeration, but the rate of increase was reduced by scale-up, which was attributed to a decrease in the liquid surface-to-volume ratio. A similar decrease in efficiency was observed in a 2000 L bioreactor by increasing the liquid volume at constant liquid surface area. The observed scale-up effects are discussed based on a simple theoretical consideration.