Viable hybridoma cells were encapsulated. The capsules were formed in one step by placing a drop of cell suspension mixed with negatively charged carboxymethylcellulose (CMC) into a positively charged chitosan solution through the interpolymeric ionic interaction between two oppositely charged polymers. These capsules were found to have a mean diameter of about 1. 5 mm and wall thickness of 3 microm. The cells grew in the capsules using supplemented DMEM/F12 (four kinds of growth factor). The maximum cell density in encapsulating cell culture reached 1 x 10(7) cells/ml, 10 times higher than that obtained in the free cell culture. The maximum monoclonal antibody concentration in the free cell culture was 15 microg/mL, but that in the capsule was 45 microg/mL The antibody produced by the cell was concentrated about four times higher inside than outside of the capsules.
We investigated the effect of dietary phospholipid (PL) concentrate from bovine milk on the epidermis. Thirteen-week-old hairless male and female mice (Hos:HR-1) were separated into two experimental groups, each fed two experimental diets: the control group and the PL group. The mice were given the experimental diets for 6 weeks. Stratum corneum hydration and transepidermal water loss (TEWL) were measured using Corneometer CM825 and Tewameter TM300 (Courage and Khazaka Electronics, Cologne, Germany) at 3 weeks and 6 weeks. After the feeding period, ceramides in stratum corneum were analyzed. We found that stratum corneum hydration and ceramides in the PL group were significantly higher than those in the control group and that TEWL in the PL group tended to decrease.These results indicate that dietary PL concentrate improves epidermal function by increasing the amount of ceramides, resulting in higher hydration.
We previously found that dietary sphingomyelin (SPM) concentrate from bovine milk improved epidermal function. In this study, we investigated the dosage of dietary SPM concentrate from bovine milk in relation to the improvement of epidermal function. Thirteen-week-old hairless male mice were separated into four experimental groups, each fed one of four types of experimental diet: the control group, the low SPM group, the medium SPM group and the high SPM group. The mice were each fed the experimental diet for 6 weeks. The stratum corneum hydration and the transepidermal water loss (TEWL) were measured using a Corneometer and a Tewameter at 3 weeks and 6 weeks. After the feeding period, ceramides in the stratum corneum were analyzed. We found that the stratum corneum hydration in all the SPM groups was significantly higher than that in the control group, whereas TEWL in all the SPM groups was significantly lower than that in the control group. Ceramides increased significantly in mice fed the medium SPM diet and statistically tended to increase in mice fed the high SPM diet. Our results indicate that a daily intake of 17 mg SPM concentrate is enough to improve epidermal function in hairless mice.
An aqueous dispersion of niosomes (non-ionic surfactant vesicles) emulsified in an external oil phase forms the vesicle-in-water-in-oil (v/w/o) system described in this paper. The properties of the surfactant used to form the vesicles, the surfactant or surfactant mixture used to stabilize the emulsion and the nature of the oil phase can be changed to provide systems of different capacities for drug or antigen and different release characteristics. The same nonionic surfactant is used as the principle amphipile to form the niosomes and to stabilize the w/o emulsion, thus promoting stability by decreasing transfer of surfactant between the stabilizing monolayers and the vesicle bilayers. The in vitro release of carboxyfluoroscein and 5-fluorouracil encapsulated within the niosomes of the v/w/o system has been investigated, the nature of the oil phase and surfactant-oil interactions being important in determining the rate of solute release. Initial studies of the system in vivo, as an adjuvant for tetanus toxoid, using cottonseed oil as the external oil phase, showed enhanced immunological activity over the free antigen or vesicles.
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