The Center for Alternatives to Animal Testing is an academic center affiliated with the Division of Toxicological Sciences in the Department of Environmental Health Sciences of the Johns Hopkins University Bloomberg School of Public Health.

 

Johns Hopkins School of Public Health

Research Grants 1983-1984

An In Vitro Epithelium Connective Tissue Barrier

Philip L. Sannes, PhD
Johns Hopkins School of Public Health, Baltimore, Maryland

Problem

Changes in the epithelium/connective tissue barrier permeability and the subsequent inflammatory events are primary responses to injury. The ability to study these phenomena in a controlled environment has been limited to simple, but highly effective, model systems, which, unfortunately have been somewhat deficient in their approximation of the in vivo situation. The purpose of this research is to set forth a system which brings together the advantages of several established in vitro systems which come a step closer to the in vivo model. The system being studied uses an epithelium-basement membrane-connective tissue matrix which will be grown on a gelatin-coated Millipore filter. The filter is used to separate segments of a modified Boyden chamber.

The barrier then will be characterized microscopically and tested in its effectiveness as a partition against chemical gradients, responsiveness to damage. and its usefulness in studying specific inflammatory events, such as chemotaxis. The system is unique since it has all of the critical elements of an in vivo tissue barrier, yet affords great flexibility in its potential application.

Impact of the Research

Since the system can be fitted with a dynamic flow system for changing gradients, introducing chemicals and monitoring effects on both sides of the barrier without disturbing the barrier physically, it more closely approximates the in vivo condition, and can be used to study inflammatory responses in vitro.