Skin response to immunogenic and non immunogenic material as applied to vaccine delivery and reconstructive surgery

The goal of this research was to demonstrate the immunogenic and non-immunogenic functions of the skin for transdermal vaccine delivery and reconstructive surgery, respectively.; In the first part, skin was used as a portal for transcutaneous vaccination with three model protein antigens based on its response to immunogenic substances. This vaccination was achieved using microporation as a vaccine delivery technology. Micropores (80 pores/cm2) were created on mice skin with the help of thermal energy. The permeation of bovine serum albumin though this microporated skin mounted on Franz diffusion cell was studied for 24 hours and was compared with that of intact skin. The permeation of BSA through intact skin was less and started approximately after 12 hours, while that through microporated skin was significantly higher and started approximately after 2 hours.; For in vivo studies, bovine serum albumin, hepatitis B surface antigen, and tetanus toxoid were selected as model antigens. Mice were anesthetized with intraperitoneal ketamine and Xylazine administration followed by microporation on the shaved abdominal region. Patch placed on microporated area was filled with sterile antigen solution of desired concentration and sealed. The patches were removed at definite time point as per the protocol and skin was washed. Primary immunization was followed by booster doses at week 4 and week 8 and blood samples were obtained at 0, 4, 6, 8, 10 weeks using tail vein bleeding. Serum was analyzed by ELISA for specific antibody against antigen used in that particular protocol. Effect of various factors like the concentration of antigen in patch, the time of patch application, the genetic make up of mice, the design of activator used for microporation and the type of vaccine was investigated using these model antigens. In addition, the technology was evaluated using microscopy and transepidermal water loss through micropores.; In the present study, BSA is delivered through micropores and has been shown to produce anti-BSA total Ig in a dose and time dependent fashion. For the dose levels 1, 10, and 100 mug/100 mul, the titers were 251, 12,900 and 27,000, respectively. Similar time dependency was observed in case of HBsAg immunization. Among four different mouse strains tested, BALB/c and SJL were found to be responsive to BSA immunization as compared to SKH and C57BL6. The intramuscular Hepatitis B vaccination gave significantly higher titer as compared to transdermal delivery. (Abstract shortened by UMI.)...