On the instrumented root canal wall, amorphous, irregular smear layer can be observed with Scanning Electron Microscope(SEM). The purpose of this study was to evaluate the effect of the presence or absence of smear layer on the adhesion of Staphylococcus aureus to the dentin of the root canal. Human incisors, extracted within 7 days, with no caries, no fracture, no calcification of canal, were selected. After cutting crown portion at cemento-enamel junction, root canal preparation was done by modified crown-down technique using Profile and Gates - Glidden Drill. During canal preparation, 10ml physiologic saline solution(group1&3) or 10ml 3.5% NaOCl(group2&4) was used as irrigation solution. And 10ml physiologic saline solution(group1&3) or 10ml 0.5M EDTA(group2&4) was applicated for final flush. After vertical sectioning and ethylene oxide gas sterilization, samples(group1&2) were immersed into BHIYHM broth inoculated with Staphylococcus aureus (ATCC 31153) and incubated for 3hrs at $37^{circ}C$. All samples were prepared for and observed with SEM(JEOL JSM840S). The data were analyzed by Mann-Whitney rank sum test. The conclusions are as follows ; 1. Smear layer covers entire root canal surface after root canal preparation. 2. Smear layer has been removed away and the entrances of dentinal tubules have opened widely, when applying 0.5M EDTA and 3.5% NaOCl. 3. A significantly higher number of bacteria were adhered to the root canal dentin without smear layer(p<0.0001). 4. Smear layer produced during root canal preparation impedes bacterial adhesion and colonization to dentin matrix, therefore inhibits canal reinfection.