The aim of this study was to evaluate the effect of autogenous tooth bone graft (ATBG) combined with platelet-rich fibrin (PRF) on bone healing in rabbit peri-implant osseous defects. Eighteen New Zealand rabbits were divided into 3 groups. Bone defects were prepared in each rabbit, and then an implant cavity was created in the defects. Dental implants were placed, and the peri-implant bone defects were treated with the following 3 methods: no graft material was applied in the control group, bone defects were treated with ATBG in the ATBG group, and bone defects were treated with ATBG combined with PRF in the ATBG+PRF group. After 28 days, the rabbits were sacrificed, and the dental implants with surrounding bone were removed. New bone formation and the percentage of bone-to-implant contact (BIC) were determined with histomorphometric evaluations. New bone formation was significantly higher in the ATBG+PRF group than the control and ATBG groups (P < .05). In addition, BIC was significantly higher in the ATBG+PRF group than in the control and ATBG groups (P < .05). The combination of ATBG with PRF contributed to bone healing in rabbits with peri-implant bone defects.

Implants are widely used in dentistry to restore missing teeth, but adequate thickness and height of bone are required for optimal implant therapy. However, alveolar bone thickness decreases after tooth extraction.1,2  As much of the alveolar bone loss occurs in the first year after tooth extraction,2  the immediate implant technique has been adopted to prevent bone loss. Although immediate implant placement decreases treatment time, preserves alveolar bone, and improves esthetics,35  buccal bone can resorb after immediate implant placement, and bony defects can form around the implants.1,68 

In past decades, various grafting materials have been used to treat peri-implant defects. Autogenous bone graft has been accepted as the ideal material because of its potential osteoinductive, osteoconductive, and osteogenic properties. Additionally, it stimulates healing and is not rejected by the immune system. However, autogenous bone graft has disadvantages, including bone resorption problems, limited donor sites, and an additional wound site.9,10  Therefore, allografts, xenografts, and synthetic grafts have been used as alternative graft materials.1113  As none of these graft materials has the desired osteoinductive properties, an alternative bone substitute has been sought.

Dentin, the main tooth structure, contains type I collagen, which promotes new bone regeneration.14  The chemical composition of bone is quite similar to that of dentin, consisting of approximately 70% hydroxyapatite, 20% collagen, and 10% body fluid.15  The dentin matrix also contains noncollagenous proteins, and these proteins include various growth factors that stimulate osteoinductive activity.16,17  Furthermore, like dentin, cementum also contains growth factors.18,19  Recently, tooth grafting materials that include dentin and cementum have been used to take advantage of these organic and inorganic components and growth factors.2022 

In dentistry, there has been increased use of biologic and synthetic molecules, various mediator cells, and platelet concentrates derived from blood to enhance periodontal regeneration and bone formation.23,24  Platelet-rich fibrin (PRF), an autogenous platelet concentrate developed by Choukroun et al,25  is a fibrin network involving platelets, growth factors, leukocytes, cytokines, and stem cells.26,27  Additionally, PRF provides mechanic fixation by binding with the graft particles.28  Autogenous PRF, alone or combined with different biomaterials, has been used to treat periodontal, bony, and peri-implant defects.12,29,30  The purpose of this study was to test the hypothesis that autogenous tooth bone graft (ATBG) combined with PRF can improve implant-to-bone integration and bone formation in peri-implant osseous defects.

The protocol was approved by the Institutional Animal Care and Use Committee of University (approval protocol No: 2016/03). The authors applied the ARRIVE (Animal Research: Reporting of in vivo Experiments) guidelines.

Animals and study design

This study was designed for 18 adult male New Zealand White rabbits weighing between 3 and 3.5 kg. For sample-size calculation, we used new bone formation results from a reference study.30  They found large effect size (f = 3.4) for new bone formation according to the independent group comparisons results. We used a lower effect-size level (f = 1) with power of 90% and a significance level of .05 for sample-size calculation, and we calculated that we needed at least 6 rats in each group. Therefore, we included 6 rats for each group (total of 18 rats) to study. In the present study, we found that the effect size for new bone formation results was f = 0.84 and this study reached 84.5% power with a 95% confidence level according to the effect size.

The animals were kept in a specially designed room and fed ad libitum on a standard diet and water. The rabbits were placed in appropriate cages in a room with an ideal daylight/darkness cycle and an ambient temperature. The animals were randomly assigned to 1 control and 2 experimental groups (n = 6 animals) and a statistical software program (SPSS Statistics v24.0; SPSS Inc, Chicago, Ill) was used for randomization. The osseous defects were created, and then implants were placed. The defects were treated with 3 methods: defects were empty around implants, defects were filled with only ATBG, and defects were filled with ATBG combined with PRF (1:1 ratio). All analyses were applied by 2 independent, calibrated examiners to determine the groups.

PRF preparation

Before the surgery, 5 mL of blood was collected in a blood collection tube without anticoagulant from the central auricular artery of each sedated animal. The blood was centrifuged immediately (PRF: 2700 rpm for 12 minutes), and a fibrin clot formed and was extracted from the tube with forceps under sterile conditions. The PRF clot was cut into small pieces and combined with the ATBG.

ATBG preparation

A mandibular left first molar tooth (included as enamel, dentin, cementum, and pulp) was removed and placed in the sterile chamber of a newly designed Smart Dentin Grinder (KometaBio, Fort Lee, NJ) and was ground and prepared for 300-μm to 1200-μm grafting particles. The particles were collected and stored in basic alcohol (0.5 M NaOH and 20% alcohol) for 10 minutes to defat and dissolve all the organic debris and bacteria.22  The solution was then drained, and the graft particles washed twice in sterile phosphate-buffered saline before being used as graft.22 

Surgical procedures

All surgeries were performed under general anesthesia with 2% xylazine (Rompun 2%, Bayer, Istanbul, Turkey) and 1% ketamine (Ketalar, Eczacibaşi-Warner Lambert, Istanbul, Turkey). The site was shaved and cleaned with povidone-iodine. After an incision was made, the tibia was exposed by subperiosteal dissection. Bone defects (10-mm diameter, 4-mm depth) were created with a trephine drill under irrigation with pure saline solution. Implant cavities (3-mm diameter, 6-mm depth) were prepared in the center of each defect according to the recommendation of the implant system manufacturer. Then, the implant cavities were rinsed with pure saline solution, dental implants (NR Line, 3.0 × 10 mm, Dentium, Cypress, Calif) were placed (6-mm depth), and primary stabilization was controlled. The upper parts of the dental implants were isolated in the center of the defects. Cover screws were placed on the implants, and the peri-implant defect was grafted with ATBG and ATBG+PRF in the experimental groups (Figure 1). After the surgery, the tissues were tightly sutured in 2 layers with degradable sutures (Pegelak, poly [glycolide-co-lactide], Dogsan, Trabzon, Turkey). Postoperatively, all the animals received Ceftriaxone 50 mg/kg (Rocephine, Deva, İstanbul, Turkey) and Carprofen 4 mg/kg (Rimadyl, Pfizer, New York, NY) intramuscularly once daily for 3 days. The animals were euthanized 28 days after surgery. The bones with the implants were dissected, and any sign of unusual healing was documented.

Figure 1.

Application of surgical procedures. (a) First right molar of rabbit. (b) Autogenous tooth bone graft (ATBG). (c) ATBG+platelet-rich fibrin (PRF). (d) Control group. (e) ATBG group, (f) ATBG+PRF group.

Figure 1.

Application of surgical procedures. (a) First right molar of rabbit. (b) Autogenous tooth bone graft (ATBG). (c) ATBG+platelet-rich fibrin (PRF). (d) Control group. (e) ATBG group, (f) ATBG+PRF group.

Close modal

Specimen preparation

The implants and surrounding bone tissue were removed en bloc and immediately immersed in formaldehyde for histologic evaluation. The specimens were dehydrated with ascending percentages of ethanol and embedded in a methylmethacrylate-based resin (Technovit 7200 VCL, Kulzer and Co, Wehrheim, Germany). All specimens were prepared with a sawing and grinding technique (Exakt Apparatebau, Norderstedt, Germany). The sections were stained with hematoxylin and eosin.

Histomorphometric analyses

All the sections were evaluated with stereologic analyses in a workstation with stereology software (Stereo Investigator version 11.0, Microbrightfield, Colchester, Vt), a charge-coupled device digital camera (Optronics MicroFire, Goleta, Calif), a personal computer, a Mac 5000 motor stage control unit (Ludl Electronic Products, Ltd, Hawthorne, NY), and a light microscope (Leica DM-4000B; Leica Microsystems, Wetzlar, Germany). A pathologist who was blinded to the animal group information, evaluated the histologic sections.

The percentage of bone-to-implant contact (BIC) was calculated by measuring the linear distance around the entire implant in which bone was in direct contact with the implant and then dividing that linear measurement by the total linear distance all the way around the implant. Additionally, histomorphometric parameters for using the calculation of new bone formation were defined as follows:

  1. Total augmented area was defined as areas including fibrovascular tissues, residual graft materials, and newly formed bone in bony defects around the implant.

  2. New bone formation (%) was calculated using the following formula: New bone formation = New bone area/Total augmented area × 100.

Statistical analyses

Shapiro–Wilk test was used to assess the normality. After parametric test assumptions were satisfied, one-way analysis of variance (ANOVA) was used for comparisons among groups. The post hoc Tukey test was used when a significant difference was determined with ANOVA. Calculations were made using the statistical software (SPSS Statistics v24.0). A difference between the groups was considered significant at P < .05. Methodology, results, and conclusions were reviewed by an independent statistician.

Wound infection or dehiscence and formation of abscesses was not detected at any surgical area. None of the animals died during the experimental procedure. All implants were in situ when the animals were sacrificed.

Newly formed bone and fibrous tissue were observed in all groups (Figure 2). However, new bone formation was lower in the control group than the test groups (P < .05). New bone formation was significantly higher in the ATBG+PRF group than the ATBG group (P < .05) (Table).

Figure 2.

Histopathological appearance of the peri-implant defect areas. (a) Control group (200 μm). (b) Higher magnification (50 μm). (c) Autogenous tooth bone graft (ATBG) group (200 μm). (d) Higher magnification (100 μm). (e) ATBG+platelet-rich fibrin group (200 μm). (f) Higher magnification (100 μm). Newly formed bone area and bone-to-implant contact are indicated with arrows.

Figure 2.

Histopathological appearance of the peri-implant defect areas. (a) Control group (200 μm). (b) Higher magnification (50 μm). (c) Autogenous tooth bone graft (ATBG) group (200 μm). (d) Higher magnification (100 μm). (e) ATBG+platelet-rich fibrin group (200 μm). (f) Higher magnification (100 μm). Newly formed bone area and bone-to-implant contact are indicated with arrows.

Close modal
Table

Percentage of direct BIC and new bone formation at defect site*

Percentage of direct BIC and new bone formation at defect site*
Percentage of direct BIC and new bone formation at defect site*

Histomorphometric evaluations revealed that BIC differed significantly among the groups (Table). BIC was significantly lower in the control group than the test groups (P < .05). Additionally, BIC was significantly higher in the ATBG+PRF group than the ATBG groups (P < .05).

The effect of ATBG combined with PRF was evaluated on peri-implant defects created in rabbit tibia. Animals, especially rabbits and rats, have been widely used to examine new bone formation. These animals are easily obtained and less expensive than other species.31  Rabbits were used in the present study because PRF and ATBG cannot be prepared from rats because of inadequate blood supply and teeth. Peri-implant defects in rabbit tibia have been used in other studies.30,32  Durmuslar et al reported that rabbit tibia has sufficient bone to create peri-implant defects.32  In the present study, the defect model was prepared as described previously.30  In this animal model, the healing of marginal defects around implants occurs after approximately 20 to 30 days.33  Hence, a 28-day period was used to evaluate the healing of peri-implant bone defects as in a previous study.30 

Peri-implant defects should be grafted to increase BIC and new bone formation. The restoration of peri-implant defects with graft materials has been investigated. Schuler et al34  reported that the use of autogenous bone grafts improved the BIC values in peri-implant defects. In another study, biphasic calcium phosphate bone substitute preserved the defect space and enhanced new bone formation.35  However, these materials have clinical disadvantages.911  Therefore, ground tooth structure has been used as a graft material in bone surgery.

The dentin matrix plays a critical role in new bone formation because of its osteoinductive properties,36  and demineralized dentin has been widely used as a bone substitute. Dentin must be demineralized for growth factors to be used because dentin tubules are expanded after demineralization, and these proteins are released.37  Reis-Filho et al17  reported that demineralized dentin matrix (DDM) stimulates new bone formation by elevating the growth factor level. Bakhshalian et al38  indicated that DDM does not cause any reaction and that it accelerates the bony repair and enhances bone quality. However, the demineralization procedure is not practical for clinical application because of the length of time required for demineralization, thus preventing the graft being performed during surgery. Additionally, Ike et al39  determined that demineralized dentin did not have osteoinductive activity, and the demineralization procedure decreases the release of growth factors, including bone morphogenetic protein-7.40  For these reasons, autogenous mineralized tooth structure was used as the bone substitute instead of DDM. Binderman et al22  reported that demineralized dentin forms inadequate bone for implant support and showed that mineralized dentin allows early loading in implant treatment because it adheres to the newly formed bone. Additionally, a mineralized tooth graft may have a beneficial effect on bone formation in implant treatment with a sinus lift operation. Autogenous tooth graft materials can be used as an alternative bone substitute.41  They do not transmit disease and can be combined with other graft materials and PRF membrane.41  In our study, according to the results of histomorphometric analysis, BIC and new bone formation was found to increase significantly in the ATBG group compared with the control group. These findings indicated that ATBG stimulates new bone formation around peri-implant defects, which is consistent with the results of previous studies.22,41  The increase of bone formation may be related to the release of the growth factors from the ATBG. Therefore, future studies are needed to evaluate the growth-factor levels in ATBG.

PRF has frequently been used in combination with other bone grafting materials to treat peri-implant defects. Simsek et al30  suggested that PRF combined with demineralized freeze-dried bone allograft significantly increased BIC and new bone formation in peri-implant defects. In another study, Melek and El Said42  grafted defects with a combination of tooth graft and injectable PRF and concluded that a combination of these materials supported bone fill. In the present study, PRF was used to increase the efficacy of ATBG. BIC, and new bone formation were determined to be significantly higher with PRF combined with ATBG compared with the ATBG and control groups. These findings showed that PRF induces new bone formation when used with graft materials and are consistent with previous studies. 12,30,31,42  Use of PRF may enhance bone healing by increasing the growth factor levels. Five milliliters of blood was received from all animals for standardization; however, the amount of PRF obtained from each animal and the growth factors in PRF may be different. This is a limitation of this study.

Grafting materials combined with growth factors can stimulate new bone formation but are very expensive. This study showed that using ATBG enhances new bone formation and BIC in peri-implant defects. However, teeth that have root canal fillings, extensive caries, or restorations are not suitable for obtaining the ATBG, and this is a limitation for clinical application. Additionally, there is no information about this graft resorption time and rate. Further studies are required to determine tooth graft resorption time and rate. Teeth include germ stem cells or pulpal stem cells, and these cells may be critically important for tissue engineering. Future studies are needed to evaluate the potential of this graft's stem cells. However, long preparation may be required for these procedures. Moreover, within the limits of this animal study, these findings suggest that ATBG is more useful in combination with a growth factor source such as PRF. Long-term studies, however, are needed to evaluate the effect of ATBG and PRF on bone formation in animals and humans.

Abbreviations

    Abbreviations
     
  • ANOVA:

    one-way analysis of variance

  •  
  • ARRIVE:

    Animal Research: Reporting of in vivo Experiments

  •  
  • ATBG:

    autogenous tooth bone graft

  •  
  • BIC:

    bone-to-implant contact

  •  
  • DDM:

    demineralized dentin matrix

  •  
  • PRF:

    platelet-rich fibrin

This study was funded by the Scientific and Technological Research Council Of Turkey (TUBITAK). The authors would like to thankfully acknowledge Hande Şenol for performing the statistical analyses, Department of Biostatistics, Faculty of Medicine, Pamukkale University, Denizli, Turkey.

The authors report no conflicts of interest related to this study.

1. 
Araujo
MG,
Lindhe
J.
Dimensional ridge alterations following tooth extraction. An experimental study in the dog
.
J Clin Periodontol
.
2005
;
32
:
212
218
.
2. 
Schropp
L,
Wenzel
A,
Kostopoulos
L,
Karring
T.
Bone healing and soft tissue contour changes following single-tooth extraction: a clinical and radiographic 12-month prospective study
.
Int J Periodontics Restorative Dent
.
2003
;
23
:
313
323
.
3. 
Cochran
DL,
Morton
D,
Weber
HP.
Consensus statements and recommended clinical procedures regarding loading protocols for endosseous dental implants
.
Int J Oral Maxillofac Implants
.
2004
;
19
(suppl)
:
109
113
.
4. 
Degidi
M,
Novaes
AB
Jr,
Nardi
D,
Piattelli
A.
Outcome analysis of immediately placed, immediately restored implants in the esthetic area: the clinical relevance of different interimplant distances
.
J Periodontol
.
2008
;
79
:
1056
1061
.
5. 
Paolantonio
M,
Dolci
M,
Scarano
A,
et al.
Immediate implantation in fresh extraction sockets. A controlled clinical and histological study in man
.
J Periodontol
.
2001
;
72
:
1560
1571
.
6. 
Araujo
MG,
Sukekava
F,
Wennstrom
JL,
Lindhe
J.
Tissue modeling following implant placement in fresh extraction sockets
.
Clin Oral Implants Res
.
2006
;
17
:
615
624
.
7. 
Araujo
MG,
Wennstrom
JL,
Lindhe
J.
Modeling of the buccal and lingual bone walls of fresh extraction sites following implant installation
.
Clin Oral Implants Res
.
2006
;
17
:
606
614
.
8. 
Blanco
J,
Nunez
V,
Aracil
L,
Munoz
F,
Ramos
I.
Ridge alterations following immediate implant placement in the dog: flap versus flapless surgery
.
J Clin Periodontol
.
2008
;
35
:
640
648
.
9. 
Jemt
T,
Lekholm
U.
Measurements of buccal tissue volumes at single-implant restorations after local bone grafting in maxillas: a 3-year clinical prospective study case series
.
Clin Implant Dent Relat Res
.
2003
;
5
:
63
70
.
10. 
Fellah
BH,
Gauthier
O,
Weiss
P,
Chappard
D,
Layrolle
P.
Osteogenicity of biphasic calcium phosphate ceramics and bone autograft in a goat model
.
Biomaterials
.
2008
;
29
:
1177
1188
.
11. 
Le Nihouannen
D,
Saffarzadeh
A,
Aguado
E,
et al.
Osteogenic properties of calcium phosphate ceramics and fibrin glue based composites
.
J Mater Sci Mater Med
.
2007
;
18
:
225
235
.
12. 
Agarwal
A,
Gupta
ND,
Jain
A.
Platelet rich fibrin combined with decalcified freeze-dried bone allograft for the treatment of human intrabony periodontal defects: a randomized split mouth clinical trail
.
Acta Odontol Scand
.
2016
;
74
:
36
43
.
13. 
Serrano Méndez
CA,
Lang
NP,
Caneva
M,
Ramírez Lemus
G,
Mora Solano
G,
Botticelli
D.
Comparison of allografts and xenografts used for alveolar ridge preservation. A clinical and histomorphometric RCT in humans
.
Clin Implant Dent Relat Res
.
2017
;
19
:
608
615
.
14. 
Güngörmüş
M,
Kaya
Ö.
Evaluation of the effect of heterologous type I collagen on healing of bone defects
.
J Oral Maxillofac Surg
.
2002
;
60
:
541
545
.
15. 
Bath-Balogh
M,
Fehrenbach
MJ.
Illustrated Dental Embryology, Histology, and Anatomy-E-Book
.
Amsterdam, Netherlands
:
Elsevier Health Sciences;
2014
.
16. 
Kim
YK,
Lee
J,
Um
IW,
et al.
Tooth-derived bone graft material
.
J Korean Assoc Oral Maxillofac Surg
.
2013
;
39
:
103
111
.
17. 
Reis-Filho
CR,
Silva
ER,
Martins
AB,
et al.
Demineralised human dentine matrix stimulates the expression of VEGF and accelerates the bone repair in tooth sockets of rats
.
Arch Oral Biol
.
2012
;
57
:
469
476
.
18. 
Gao
J,
Symons
AL,
Bartold
PM.
Expression of transforming growth factor-beta 1 (TGF-beta1) in the developing periodontium of rats
.
J Dent Res
.
1998
;
77
:
1708
1716
.
19. 
Schmidt-Schultz
TH,
Schultz
M.
Intact growth factors are conserved in the extracellular matrix of ancient human bone and teeth: a storehouse for the study of human evolution in health and disease
.
Biol Chem
.
2005
;
386
:
767
776
.
20. 
Jeong
H-R,
Hwang
J-H,
Lee
J-K.
Effectiveness of autogenous tooth bone used as a graft material for regeneration of bone in miniature pig
.
J Korean Assoc Oral Maxillofac Surg
.
2011
;
37
:
375
379
.
21. 
Kim
YK,
Kim
SG,
Byeon
JH,
et al.
Development of a novel bone grafting material using autogenous teeth
.
Oral Surg Oral Med Oral Pathol Oral Radiol Endod
.
2010
;
109
:
496
503
.
22. 
Binderman
I,
Hallel
G,
Nardy
C,
Yaffe
A,
Sapoznikov
L.
A novel procedure to process extracted teeth for immediate grafting of autogenous dentin
.
J Interdiscipl Med Dent Sci
.
2014
;
2
:
2
.
23. 
Fernandes
G,
Yang
S.
Application of platelet-rich plasma with stem cells in bone and periodontal tissue engineering
.
Bone Res
.
2016
;
4
:
16036
.
24. 
Gamal
AY,
Abdel Ghaffar
KA,
Alghezwy
OA.
Crevicular fluid growth factors release profile following the use of platelet-rich fibrin and plasma rich growth factors in treating periodontal intrabony defects: a randomized clinical trial
.
J Periodontol
.
2016
;
87
:
654
662
.
25. 
Choukroun
J,
Adda
F,
Schoeffer
C,
Vervelle
A.
An opportunity in perio-implantology: PRF [in French]
.
Implantodontie
.
2001
;
42
:
55
62
.
26. 
Dohan
DM,
Choukroun
J,
Diss
A,
et al.
Platelet-rich fibrin (PRF): a second-generation platelet concentrate
.
Part III: leucocyte activation: a new feature for platelet concentrates? Oral Surg Oral Med Oral Pathol Oral Radiol Endod
.
2006
;
101
:
e51
e55
.
27. 
Kang
YH,
Jeon
SH,
Park
JY,
et al.
Platelet-rich fibrin is a bioscaffold and reservoir of growth factors for tissue regeneration
.
Tissue Eng Part A
.
2011
;
17
:
349
359
.
28. 
Simonpieri
A,
Choukroun
J,
Del Corso
M,
Sammartino
G,
Dohan Ehrenfest
DM.
Simultaneous sinus-lift and implantation using microthreaded implants and leukocyte- and platelet-rich fibrin as sole grafting material: a six-year experience
.
Implant Dent
.
2011
;
20
:
2
12
.
29. 
Simonpieri
A,
Del Corso
M,
Vervelle
A,
et al.
Current knowledge and perspectives for the use of platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) in oral and maxillofacial surgery part 2: bone graft, implant and reconstructive surgery
.
Curr Pharm Biotechnol
.
2012
;
13
:
1231
1256
.
30. 
Simsek
S,
Ozec
I,
Kurkcu
M,
Benlidayi
E.
Histomorphometric evaluation of bone formation in peri-implant defects treated with different regeneration techniques: an experimental study in a rabbit model
.
J Oral Maxillofac Surg
.
2016
;
74
:
1757
1764
.
31. 
Acar
AH,
Yolcu
U,
Gul
M,
Keles
A,
Erdem
NF,
Altundag Kahraman
S.
Micro-computed tomography and histomorphometric analysis of the effects of platelet-rich fibrin on bone regeneration in the rabbit calvarium
.
Arch Oral Biol
.
2015
;
60
:
606
614
.
32. 
Durmuslar
MC,
Balli
U,
Dede
FO,
et al.
Histological evaluation of the effect of concentrated growth factor on bone healing
.
J Craniofac Surg
.
2016
;
27
:
1494
1497
.
33. 
Rossi
F,
Botticelli
D,
Pantani
F,
Pereira
FP,
Salata
LA,
Lang
NP.
Bone healing pattern in surgically created circumferential defects around submerged implants: an experimental study in dog
.
Clin Oral Implants Res
.
2012
;
23
:
41
48
.
34. 
Schuler
RF,
Janakievski
J,
Hacker
BM,
O'Neal
RB,
Roberts
FA.
Effect of implant surface and grafting on implants placed into simulated extraction sockets: a histologic study in dogs
.
Int J Oral Maxillofac Implants
.
2010
;
25
:
893
900
.
35. 
Kim
S,
Jung
U,
Lee
YK,
Choi
SH.
Effects of biphasic calcium phosphate bone substitute on circumferential bone defects around dental implants in dogs
.
Int J Oral Maxillofac Implants
.
2011
;
26
:
265
.
36. 
Gomes
MF,
Banzi
EC,
Destro
MF,
Lavinicki
V,
Goulart
M.
Homogenous demineralized dentin matrix for application in cranioplasty of rabbits with alloxan-induced diabetes: histomorphometric analysis
.
Int J Oral Maxillofac Implants
.
2007
;
22
:
939
947
.
37. 
Li
R,
Guo
W,
Yang
B,
et al.
Human treated dentin matrix as a natural scaffold for complete human dentin tissue regeneration
.
Biomaterials
.
2011
;
32
:
4525
4538
.
38. 
Bakhshalian
N,
Hooshmand
S,
Campbell
SC,
Kim
JS,
Brummel-Smith
K,
Arjmandi
BH.
Biocompatibility and microstructural analysis of osteopromotive property of allogenic demineralized dentin matrix
.
Int J Oral Maxillofac Implants
.
2013
;
28
:
1655
1662
.
39. 
Ike
M,
Urist
MR.
Recycled dentin root matrix for a carrier of recombinant human bone morphogenetic protein
.
J Oral Implantol
.
1998
;
24
:
124
132
.
40. 
Pietrzak
WS,
Ali
SN,
Chitturi
D,
Jacob
M,
Woodell-May
JE.
BMP depletion occurs during prolonged acid demineralization of bone: characterization and implications for graft preparation
.
Cell Tissue Bank
.
2011
;
12
:
81
88
.
41. 
Pohl
V,
Schuh
C,
Fischer
MB,
Haas
R. A
New method using autogenous impacted third molars for sinus augmentation to enhance implant treatment: case series with preliminary results of an open, prospective longitudinal study
.
Int J Oral Maxillofac Implants
.
2016
;
31
:
622
630
.
42. 
Melek
LN,
El Said
MM.
Evaluation of “autogenous bioengineered injectable PRF–tooth graft” combination (ABIT) in reconstruction of maxillary alveolar ridge defects: CBCT volumetric analysis
.
Saudi J Dent Res
.
2017
;
8
:
86
96
.