Comparison of the Proliferation pattern of Cultured Rat Calvaria Cell on the Resorbable Barrier Membrane

Lee, Chang Hoon; Lee, Man Sup; Kwon, Young Hyuk; Park, Joon Bong; Herr, Yeek

Abstract

The purpose of this study is to evaluate the phenomenon of attachment and spreading of the cultured rat calvarial cell inoculated on their surface of different kinds of biodegradable membrane which had been used on tissue regeneration on periodontal defects by using scanning electron microscope. In this experiment 30 Sprague-Dawley male rats (mean BW 150gm) were used to harvest abundant number of cell in the short period. The rats were sacrificed by decapitatioan to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Biodegradable barrier membrane were collected with collagen type, and were divided into 3 different kind of surface such as scattered, polarized and fine-net type as their surface texture. Microcover plate which usually used for cell cuture was used as control for smooth surface. All the membrane were seeded with cultured calvarial cell on their surface. The number of cell inoculated on the membrane were 1x106 Cells/ml. After the culture as designed time, all the membrane were washed with 0.1 M Phosphate Buffered saline and fixed with 2.5% Glutaraldehyde. And all specimen were treated with OsO4, and Tannic acid before drying the cell for coating the cell with gold. Scanning Electron Microscope was used to observation. The following results were obtained. 1. During the whole period of experiment, the phenomenon of cell attachment and spreading were revealed similar pattern to compare with smooth surface culture plate and ordinary culture dish. 2. The shape of cell attchment and spreading on the surface of barrier membrane were observed no remaked difference pattern between smooth surface culture plate and ordinary culture dish. 3. The cytoplasmic process of cultured calvaria cell extent to the deep portion of barrier membrane like as their own proper shape. 4. There were no remarkable relationships between the degree of cultured cell spreading and surface structure of barrier membrane. 5. Slight starified layer of cultured calvaria cell were observed on the scattered type of resorbable membrane. Conclusively, this study thus suggest that cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of carrier for many cell which could be used as new tissue regeneration, and those tissue engeering technique may become an new method in the approach to the repair of bone defects.