In vitro analysis of particle penetration of smokeless tobacco forms using egg shell membrane as a substrate

Aims: The aim of the following study is to determine the particle sizes of smokeless tobacco forms and thereby evaluate the degree of diffusion of the products using an egg shell membrane as a natural substrate by scanning electron microscopy (SEM). Subjects and Methods: The particle size of smokeless tobacco forms namely mawa, gutka, khaini, and tobacco leaves was determined by image analysis and the products were subjected on an egg shell membrane subjected to artificial saliva along with constant grinding force. The processed egg shell membrane was then examined by SEM to evaluate the morphology and the degree of diffusion of these particles. Results: The morphometric image analysis revealed khaini to be smallest in size followed by mawa, gutka, and tobacco leaves. The control group (egg shell membrane not subjected to any products) under SEM demonstrated intricately woven collagen meshwork, which was regular in length, branched and anastamosed with each other. Khaini exposed membrane demonstrated maximum particle diffusion with disruption of collagen meshwork. Mawa exposed membranes demonstrated minimal particle penetration which were adherent to the collagen meshwork which was irregular with increased interfibrillar space. The egg shell membrane exposed to gutka demonstrated particle penetration at higher magnification with more or less a regular collagen meshwork. The tobacco leaf exposed egg shell membrane demonstrated minimal blebs on the meshwork surface. Conclusions: The degree of diffusion gradually increased with a decrease in the particle size establishing an inverse relationship.