autogenous bone grafts are extensively used to augment severely resorbed alveolar ridges , although other materials , such as , undecalcified , allogeneic devitalized bone , frozen bone , processed xenogeneic bone , deproteinized bone , and hydroxyapatite ( ha ) crystals , are also used for bone regeneration . initially , particulate dentin ( tooth ash ) was used as a graft material , because hydroxyapatite is the main ingredient of teeth . because it is bioresorbable , particulate tooth ash can also be used as an alternative graft material for implants . it is composed of a hydrated organic matrix consisting mainly of type i collagen and an inorganic reinforcing phase of carbonated apatite . dentin is composed of inorganic ( 70% 75% ) , organics ( 20% ) , and water ( 10% ) , whereas in alveolar these percentages are 65% , 25% , and 10% , respectively . on the other hand , the bone - inducing property of dentin was discovered in 1967 when demineralized dentin matrices ( ddm ) of rabbit were found to induce bone formation in intramuscular tissues . a problem with demineralized dentin as a graft material is the time it takes to decalcify and produce as a bone graft material from teeth . in particular , tooth blocks require more processing time than tooth chips or powder . the conventional method requires 10 days or more to complete because bone transplant material must be prepared using extracted teeth . this requires transport to a processing company , and at least 10 hours is required to demineralize a tooth block . as a faster alternative method , we developed a vacuum - ultrasonic accelerator ( vacuasonic ; cosmobio - medicare , seoul , korea ) based method that can be performed in clinics . the whole process can be completed within two hours regardless of the form ( powder , chip or block ) used . ultrasonication uses for cleaning , but because of the hardness of teeth , it is difficult to clean hidden surfaces ( dentinal tubules ) . the chemical treatment employed with ultrasonication included a vacuum , which noticeably improved efficiency , and markedly reduced overall treatment time . we studied the ultra - structure and chemical components of newly processed tooth graft material by scanning electron microscopy ( sem ) and energy dispersive x - ray spectroscopy ( eds ) . a clinical case revealed the ability of the graft materials produced to regenerated bone after implant surgery . here we report preliminary results obtained from our experimental studies and a clinical case of alveolar bone regeneration using the novel tooth grafts . chungbuk national hospital biobank provided tooth samples ( n=8 ) for this research , and all donors provided informed consent and the study was approved by irb . the samples were randomly allocated to an autogenous demineralized dentin ( add ) group ( n=5 ) and an untreated group ( n=3 ) . human extracted teeth were washed in a solution of 0.9% nacl after soft tissue and pulp removal and stored at 70c . in the add group , remnant tissues around the roots of extracted teeth were removed using a surgical blade or periosteal elevator . a dental handpiece was used to remove pulp tissue from the pulp chamber and root canals and perfuse the reagent effectively . in addition 0.3 mm diameter holes were made along the entire tooth surface using a 330 bur ( fig . the tooth was placed in a sterile solution for 10 minutes in the vacuum ultrasonic system . teeth were demineralized in a 0.6 n solution of hcl for 90 minutes in the vacuum ultrasonic machine ( vacuasonic ) , and washed for twice for 15 minutes in deionized sterile water or phosphate buffered saline ( pbs ) solution . remnant tissues around the roots were removed using a surgical blade or periosteal elevator , then the teeth were washed in a saline solution . two groups of tooth samples were prepared in the form of 2 mm cubes for sem and eds studies . cubes were removed from the tooth necks with a # 15 blade ( add ) and a high speed bur ( untreated teeth ) . to quantify weight reduction , weights were measured before and after processing in add group . observations of add material and untreated tooth surfaces were performed optically using a scanning electron microscope , in secondary electron mode . sem studies were performed using a nanoscope iiia instrument ( digital instruments , tonawanda , ny , usa ) , after coating samples with a gold - palladium alloy . to determine whether a change in mineral components occurred during processing , eds was performed using the team system ( bruker co. , ewing , nj , usa ) . to determine internal mineral concentrations , processed tooth materials and unprocessed teeth were checked for calcium concentration ( wt% ) at a depth of 300 to 600 micrometers from tooth surface . a 45-year - old male patient visited chungbuk national university hospital for extraction of # 48 tooth . radiological and clinical examinations revealed an impacted third molar , a periapical and a periodontal lesion of # 47 , and an edentulous alveolar ridge on # 44 with narrow ridge ( fig . 3 ) . therefore , surgical extraction of # 47 and 48 and implant placement on # 44 and 47 with add were planned . # 47 and 48 teeth were extracted before surgery , and block - type autogenous tooth graft materials were produced in a clinic . for personal reasons , surgery was delayed for six months , and processed materials were stored at 70c in a freezer . significant narrow ridges on # 44 ( 2 mm ) and # 47 ( 4 mm ) edentulous sites were noted . implants were placed on the # 44 ( diameter 4.0 mm , length of 10 mm ; tsiii ; osstem , seoul , korea ) and # 47 area diameter 4.5 mm , length of 10 mm ; tsiii ) . an autogenous tooth chip was prepared with the bone mill and placed on the buccal side of alveolar ridge . to increase volume , a block type of add was transplanted upwards autogenous tooth chip with resorbable collagen membrane ( cyaddlast ; osteogenic biomedical inc . , lubbock , tx , usa ) ( fig . 6 ) . after suturing without tension , antibiotics and anti - inflammatory analgesic were administered and the patient gargled with saline . chungbuk national hospital biobank provided tooth samples ( n=8 ) for this research , and all donors provided informed consent and the study was approved by irb . the samples were randomly allocated to an autogenous demineralized dentin ( add ) group ( n=5 ) and an untreated group ( n=3 ) . human extracted teeth were washed in a solution of 0.9% nacl after soft tissue and pulp removal and stored at 70c . in the add group , remnant tissues around the roots of extracted teeth were removed using a surgical blade or periosteal elevator . a dental handpiece was used to remove pulp tissue from the pulp chamber and root canals and perfuse the reagent effectively . in addition 0.3 mm diameter holes were made along the entire tooth surface using a 330 bur ( fig . the tooth was placed in a sterile solution for 10 minutes in the vacuum ultrasonic system . teeth were demineralized in a 0.6 n solution of hcl for 90 minutes in the vacuum ultrasonic machine ( vacuasonic ) , and washed for twice for 15 minutes in deionized sterile water or phosphate buffered saline ( pbs ) solution . remnant tissues around the roots were removed using a surgical blade or periosteal elevator , then the teeth were washed in a saline solution . two groups of tooth samples were prepared in the form of 2 mm cubes for sem and eds studies . cubes were removed from the tooth necks with a # 15 blade ( add ) and a high speed bur ( untreated teeth ) . to quantify weight reduction , weights were measured before and after processing in add group . observations of add material and untreated tooth surfaces were performed optically using a scanning electron microscope , in secondary electron mode . sem studies were performed using a nanoscope iiia instrument ( digital instruments , tonawanda , ny , usa ) , after coating samples with a gold - palladium alloy . to determine whether a change in mineral components occurred during processing , eds was performed using the team system ( bruker co. , ewing , nj , usa ) . to determine internal mineral concentrations , processed tooth materials and unprocessed teeth were checked for calcium concentration ( wt% ) at a depth of 300 to 600 micrometers from tooth surface . a 45-year - old male patient visited chungbuk national university hospital for extraction of # 48 tooth . radiological and clinical examinations revealed an impacted third molar , a periapical and a periodontal lesion of # 47 , and an edentulous alveolar ridge on # 44 with narrow ridge ( fig . 3 ) . therefore , surgical extraction of # 47 and 48 and implant placement on # 44 and 47 with add were planned . # 47 and 48 teeth were extracted before surgery , and block - type autogenous tooth graft materials were produced in a clinic . for personal reasons , surgery was delayed for six months , and processed materials were stored at 70c in a freezer . significant narrow ridges on # 44 ( 2 mm ) and # 47 ( 4 mm ) edentulous sites were noted . implants were placed on the # 44 ( diameter 4.0 mm , length of 10 mm ; tsiii ; osstem , seoul , korea ) and # 47 area diameter 4.5 mm , length of 10 mm ; tsiii ) . an autogenous tooth chip was prepared with the bone mill and placed on the buccal side of alveolar ridge . to increase volume , a block type of add was transplanted upwards autogenous tooth chip with resorbable collagen membrane ( cyaddlast ; osteogenic biomedical inc . , after suturing without tension , antibiotics and anti - inflammatory analgesic were administered and the patient gargled with saline . the weights of teeth before processing were measured at an average 0.38 g ( range , 0.230.53 g ) . the weights of teeth after processing averaged 0.20 g ( range , 0.130.32 g ) , 43.4% average reduction ( table 1 ) . fig . 8 shows a dentin surface with a smear layer and ground tooth particles produced by a high speed bur . in untreated teeth smear layers were composed of small particles of dentin and dendrites covered dentin surface . the calcium concentration of add was 6.52 wt% whereas that of normal dentin was 31.17 wt% at a depth of 300600 micrometers from external surfaces ( table 2 ) , indicating that 79% of the calcium had been removed ( fig . no significant complications , such as , infection , uncontrolled dehiscence , graft failure , or implant failure occurred . further examination of computer tomographic views was performed to confirm bone healing at add treated defect sites ( fig . the weights of teeth before processing were measured at an average 0.38 g ( range , 0.230.53 g ) . the weights of teeth after processing averaged 0.20 g ( range , 0.130.32 g ) , 43.4% average reduction ( table 1 ) . 8 shows a dentin surface with a smear layer and ground tooth particles produced by a high speed bur . in untreated teeth smear layers the calcium concentration of add was 6.52 wt% whereas that of normal dentin was 31.17 wt% at a depth of 300600 micrometers from external surfaces ( table 2 ) , indicating that 79% of the calcium had been removed ( fig . no significant complications , such as , infection , uncontrolled dehiscence , graft failure , or implant failure occurred . further examination of computer tomographic views was performed to confirm bone healing at add treated defect sites ( fig . the present study documented the characteristics and the ability of alveolar bone reconstruction using rapidly demineralized dentin ( add ) as a bone graft material . dentin is an acellular collagen - rich tissue matrix without vessels , whereas bone is a cellular tissue with vessels . however , dentin and bone have similar compositions , that is , 10% body fluid , 20% organic materials , and 70% minerals ( mainly ha ) , and contain bone morphogenetic proteins ( bmps ) that have the ability to induce bone and cartilage in formation in non - skeletal sites . the major component of the inorganic material of teeth contains several types of calcium phosphate including ha . earlier research found that tooth ash can be used as graft material for the reconstruction of bone . demineralization of dentin increases the bio - availability of matrix - associated non - collagenous proteins , rendering these grafts osteoinductive . non - collagenous dentin proteins , such as , osteocalcin , osteonectin , phosphoprotein , and sialoprotein are involved in bone calcification . furthermore decalcified dentine is penetrated and resorbed more slowly than decalcified bone , because it is denser and has neither old vascular channels nor marrow spaces . for the same reason , the decalcification time of teeth is greater than that of bone . long processing times for teeth are problematic clinically . moreover , excessive acid exposure might diminish bmp retention due to diffusional losses and graft potency . as described by pietrzak et al . , the demineralization of bovine cortical bone using 0.5 n hcl occurred within 90 minutes and peak bmp-7 levels were reached at that point , and then fell . to overcome this problem sem analysis of newly decalcified dentin ( add ) revealed dentinal tubules , which we believe act as a network for the diffusion of nutrients after grafting . type i collagen fiber accounts for 90% of the organic matrix , and is important in calcification . ultrasonic treatment creates cavitation , a high energy phenomenon , inside an object by repeated compression and expansion . when this cavitation is present on the surface of an object , the energy released activates a reaction between the reagent and the object . in order to use the ultrasonic system , we considered two factors ; temperature and air bubbles . increased temperature during processing affects the preservation of protein substance in graft material . as air bubbles in dentinal tubules hinders rapid demineralization , the devised method facilitates clinical application because it can perform block processing within two hours . if the operator chooses to use chip or powder , changes can be made during bone milling . in the described case furthermore , our high - resolution imaging and eds studies confirmed the decalcification of dentin . in the clinical case , we observed good bone formation in narrow alveolar ridges . finally , we conclude rapid preparation of autogenous teeth using the devised method makes one - day grafting possible .
purpose : the current gold standard for clinical jawbone formation involves autogenous bone as a graft material . in addition , demineralized dentin can be an effective graft material . 
 although demineralized dentin readily induces heterotopic bone formation , conventional decalcification takes three to five days , so , immediate bone grafting after extraction is impossible . 
 this study evaluated the effect of vacuum ultrasonic power on the demineralization and processing of autogenous tooth material and documented the clinical results of rapidly processed autogenous demineralized dentin ( add ) in an alveolar defects patient.methods:the method involves the demineralization of extracted teeth with detached soft tissues and pulp in 0.6 n hcl for 90 minutes using a heat controlled vacuum - ultrasonic accelerator . 
 the characteristics of processed teeth were evaluated by scanning electron microscopy ( sem ) , and energy dispersive x - ray spectroscopy ( eds ) . 
 bone grafting using add was performed for narrow ridges augmentation in the mandibular area.results:the new processing method was completed within two hours regardless of form ( powder or block ) . 
 eds and sem uniformly demineralized autotooth biomaterial . 
 after six months , bone remodeling was observed in augmented sites and histological examination showed that add particles were well united with new bone . 
 no unusual complications were encountered.conclusion:this study demonstrates the possibility of preparing autogenous tooth graft materials within two hours , allowing immediate one - day grafting after extraction .