The purpose of this study was to compare the effect of tetracycline HCL, Citric acid and PrefGel applied on the root surfaces that planed with periodontal curret with Roto bur. In this study, 20 extracted teeth with advanced periodontal disease were used. The teeth were root planing with periodontal curette and Roto bur. Following root planing, each agents was burnished on the prepared root surface for 3 minutes to find opened dentinal tubules. And then, each specimens were investigated using scanning electron microscope. Amount of remained cementum by loss of tooth substance index and the number of opened dentinal tubules were evaluated to each specimens The results were as follows. 1. Groups treated with periodontal curette were almost seemed no removed. Other groups treated with Roto bur showed partially opened dentinal tubule orifices. 2. Loss of tooth substance index were compared between groups. There was no statistically difference between periodontal curette groups. Between Roto bur groups was alike. But there were statistically differences between periodontal curette and Roto bur groups. 3. At comparing with various root conditioning agents, Tetracycline HCL group took statistically higher than Citric acid and PrefGel in opened dentinal tubules. On the other hand, there was no statistically difference between Citric acid group and PrefGel group. As a result of this study, groups treated with Roto bur showed more cementum removed than groups treated with periodontal curette. In a treatment for regeneration of periodontal tissue, it was regarded that Roto bur should be used and that Tetracycline HCL would be more effective as chemical root conditioning agent.