To analyze physicochemical such as surface structures, the crystallinity, chemical composition, calcium phosphate dissolution and osteogenic properties of tooth derived bone substitute (TDBS) processed chair-side and other grafting materials. The number of anaerobic and facultative anaerobic bacteria in the supernatant of processed TDBS was determined. Human osteoblasts were cocultured with TDBS or allograft in transwell system to examine cell migration. BMP2 released from TDBS was measured by ELISA. TDBS had high crystallinity similar to BoneCeramic while it had a broad pattern to ramus bone, OraGRAFT, and Bio-Oss. TDBS contained carbon, calcium, oxygen, phosphate, sodium and magnesium elements like others. Calcium/phosphorus dissolution of TDBS show closely related to those of mandibular ramus bone and OraGRAFT. In addition, microbial decontamination of TDBS by the chemical processing revealed a hundred percent efficacy. The osteoconductive and osteoinductive properties demonstrated in the TDBS processed chairside suggested the potential of an alternative for bone grafting material.