Periodontal regeneration through the use of root demineralization received a lot of interest in periodontology. Topical application of acid to dentin surfaces produced a zone of demineralization, exposing dentin collagen fibrils and opening dentin tubules. In this study, the in vitro effects of different tetracycline HCl application techniques were investigated. According to the results of this SEM study, it may be desirable to apply tetracycline HCl using burnishing technique to expose maximum intertubular fibrils and for the tubular openings. However, this technique should be studied when placed in an in vivo system.