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International Journal of Dental and Health Sciences
Volume 01, Issue 05Case Report
EXTRACTION AND IMMEDIATE IMPLANT
PLACEMENT USING A COMBINED PRF,
AND PROVISIONALIZATION TECHNIQUE
Abu-Hussein Muhamad1
,Abdulgani Azzaldeen2
1.University of Naples Federic II, Naples, Italy, Department of Pediatric Dentistry, University
of Athens, Athens, Greece.
2. Department of Conservative Dentistry, Al-Quds University, Jerusalem, Palestine.
ABSTRACT:
The placement of dental implant into fresh extraction sockets was introduced in
1970 and is a well established treatment option for replacing missing teeth, allowing the
restoration of masticatory function, speech and esthetics. Immediate post extraction
implant placement is a well accepted protocol because of shorter total treatment time,
maintenance of socket wall, reduced operative time and better actual implant placemen1
. In
immediate implant placement there is gap present between implant surface and socket wall
and there are various materials used to fill this gap for better osseointegration but these
materials are either expensive or not so effective. Platelet- rich fibrin (PRF) is simple,
natural and inexpensive. Platelet-rich fibrin(PRF) concentrate helps in healing process,
osteoblastic activity, angiogenesis, release growth factors and able to stimulate defense
mechanism. PRF has been used as graft material in sinus lift procedure with simultaneous
implant with success. This article describes a case in which the fenestration defect around
an implant was treated by the application of platelet rich fibrin, a second generation platelet
concentrate along with bone graft, and guided tissue regeneration membrane.
Key words: Guided bone regeneration, growth factors, platelet rich fibrin
INTRODUCTION:
Teeth replacement using dental implants
has proven to be a successful and
predictable treatment procedure;
different placement and loading protocols
have evolved from the first protocols in
order to achieve quicker and easier
surgical treatment times [1]
. In situations
where a tooth requires extraction and
replacement, original protocol (gold
standard) suggested a 6-12 month waiting
period before implant placement [2]
. The
original protocol has been challenged
within the last decade and new protocols
have been developed in which implants
are placed at the time of extraction of the
tooth. This protocol where implants have
been placed at the time of tooth
*Corresponding Author Address: Abdulgani Azzaldeen,Department of Conservative Dentistry, Al-Quds University,
Jerusalem, Palestine. Email: abdulganiazzaldeen@gmail.com
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
extraction is known as immediate
implants[3]
.
When an implant is placed in a fresh or
recent extraction alveolus a gap between
the implant surface and the bone walls of
the socket may occur. The presence or
size of the gap is both influenced by the
configuration of the alveolus and by the
design and width of the implant [4]
. Wilson
et al.[8]
and Paolantonio et al.[9]
demonstrated that for implants with a
horizontal defect of 2 mm or less,
spontaneous bone healing and
osseointegration take place if the implant
has a rough surface [5,6]
. Horizontal defects
in excess of 2 mm have been shown to not
heal predictably with bone[5]
. So To
compensate for these problems, guided
bone regeneration (GBR) using autografts,
allografts, or alloplasts; barrier
membranes; or combination therapy has
been accomplished with what appear to
be successful clinical results [3,7]
.
Implant therapy based on the principle of
osseointegration has seen a remarkable
expansion of its application in dentistry, in
recent years. In the last decade, dental
implants have become a reliable
procedure for the treatment of partially or
completely edentulous jaws. The lack of
bone adjacent to an implant can be
considered a true "bony defect" and
several techniques have been proposed to
promote defect fill with newly formed
bone. One of the most popular
procedures is guided bone regeneration
(GBR), which involves placing a membrane
over the defect to create a secluded space
into which osteogenic cells can migrate
and remain undisturbed over the exposed
part of the implant[7,8]
.
Short-term animal and human studies
have shown these immediate implants to
be comparable to implants placed into
healed bone. The advantage of the
procedure include fewer surgical sessions,
elimination of the waiting period for
socket healing, shortened edentulous
time period, reduced overall cost, as well
as preservation of bone height and width
[2,9]
. Although immediate implantation is
more demanding both surgically and
prosthetically compared to the
conventional placement technique, the
advantages make it very appealing to
patients who are in need of both
extractions and implant therapy.[1,3,7,10]
Platelet-rich plasma (PRP) is an
autologous product that concentrates a
large number of platelets in a small
volume of plasma. PRP functions as a
fibrin tissue adhesive with hemostatic and
tissue sealing properties, but it differs
from fibrin glue and other platelet-poor
tissue adhesives because its platelets
provide a unique ability to promote
wound healing and enhance
osteogenesis[1,2]
.
PRP provides an immediate surgical
hemostatic agent that is biocompatible,
safe, and effective. PRP accelerates
endothelial, epithelial, and epidermal
regeneration, stimulates angiogenesis,
enhances collagen synthesis, promotes
soft tissue healing, decreases dermal
scarring, enhances the hemostatic
response to injury, and reverses the
inhibition of wound healing caused by
175
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
glucocorticoids. The high leukocyte
concentration of PRP has an added
antimicrobial effect. Since PRP is an
autologous blood product, it carries no
risk of transmitting infectious disease.[4,5,6]
PRP has an extremely broad range of
clinical healing applications in head and
neck surgery, otolaryngology,
cardiovascular surgery, burns and wound
healing, oral and maxillofacial surgery,
cosmetic surgery, and periodontics In
addition to its effectiveness for patients
with chronic non-healing wounds, it has
also been used as an antiangiogenic agent
and as a carrier for growth factors.[1,3,4]
In surgical settings, PRP decreases the
frequency of intraoperative and
postoperative bleeding at donor and
recipient sites, accelerates soft-tissue
healing, supports the initial stability of
grafted tissue at recipient sites as a result
of its cohesive and adhesive nature,
promotes rapid vascularization of healing
tissue by delivering growth factors and,
when used in combination with bone
replacement materials, induces
regeneration.[3]
The term tissue engineering was originally
coined to denote the construction in the
laboratory of a device containing viable
cells and biologic mediators (e.g., growth
factors and adhesins) in a synthetic or
biologic matrix, which could be implanted
in patients to facilitate regeneration of
particular tissues. The role of tissue
oxygenation in wound healing became the
focal point in the 1980s. Tissue
oxygenation enhances phagocytic and
bactericidal ability of host immune cells
and supports collagen as well as other
protein synthetic events. The importance
of growth factors in enhancing wound
healing has become the focus of research
in the present day. In addition, a link has
been established between tissue
oxygenation and growth factors.
Macrophage stimulation causes the
release of angiogenic and other growth
factors that support wound healing and
resist infection. In general, tissue
engineering combines three key elements,
namely scaffolds (collagen, bone mineral),
signaling molecules (growth factors), and
cells (osteoblasts, fibroblasts). Tissue
engineering has been redefined presently
as the relatively new, highly promising
field of reconstructive biology, which
draws on the recent advances in medicine
and surgery, molecular and cellular
biology, polymer chemistry, and
physiology.[4,5,6]
Platelets isolated from peripheral blood
are an autologous source of growth
factors. When platelets in a concentrated
form are added to graft materials, a more
predictable outcome is derived. Platelet-
rich plasma (PRP) is an easily accessible
source of growth factors to support bone-
and soft-tissue healing. It is derived by
methods that concentrate autologous
platelets and is added to surgical wounds
or grafts and to other injuries in need of
supported or accelerated healing. A blood
clot is the center focus of initiating any
soft-tissue healing and bone regeneration.
In all natural wounds, a blood clot forms
and starts the healing process. PRP is a
simple strategy to concentrate platelets or
enrich natural blood clot, which forms in
176
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
normal surgical wounds, to initiate a more
rapid and complete healing process. A
natural blood clot contains 95% red blood
cells, 5% platelets, less than 1% white
blood cells, and numerous amounts of
fibrin strands. A PRP blood clot contains
4% red blood cells, 95% platelets, and 1%
white blood cells[2,7,9]
.
The use of PRP in place of recombinant
growth factors has several advantages, in
that growth factors obtained from
platelets not only have their own specific
action on tissues but also interact with
other growth factors, resulting in the
activation of gene expression and protein
production. Therefore, the properties of
PRP are based on the production and
release of multiple growth and
differentiation factors upon platelet
activation. These factors are critical in the
regulation and stimulation of the wound
healing process, and they play an
important role in regulating cellular
processes such as mitogenesis,
chemotaxis, differentiation, and
metabolism.
Platelet rich fibrin, which is a second
generation platelet concentrate, offers
the surgeon an access to growth factors
with a simple and available technology.
These growth factors which are
autologous, nontoxic and non
immunogenic, enhance and accelerate the
normal bone regeneration pathways.
[10,11,12]
This article presents a case report in which
a fenestration defect around an implant
was treated by GBR with platelet rich
fibrin (PRF)-enhanced bone graft and the
PRF smeared barrier membrane for
guided bone regeneration.
CASE DETAIL:
A healthy 27-year-old male oral surgery
resident was referred to the author’s
private office for a apical lesions in the
maxillary anterior teeth need to be
removed and implant restoration . He
gave the history of trauma four months
back and simultaneous avulsion of the left
central incisor tooth. On clinical
examination , height of the alveolar ridge
was adequate but the width of the ridge
was inadequate for an implant placement.
The ridge was found to be a class I defect
according to Siebert’s classification.
Intraoral radiographs also revealed a
deficient ridge. Ridge augmentation using
bone graft with platelet rich fibrin (PRF)
was planned. The patient was clinically
healthy with no history of systemic
diseases. Informed consent was taken
from the patient.Fig.1-2
PREPARATION OF PRF:
PRF belongs to a new generation of
platelet concentrate but with a simplified
processing as compared to platelet rich
plasma (PRP). Blood was drawn into 10ml
test tubes without an anticoagulant and
centrifuged immediately for 12 minutes at
2,700 rpm .
Indication for using PRF
I. BONE REGENERATION :
1. Sinus lift grafting
2. Ridge augmentation
3. Repair of bone defects created by
removal of teeth or small cyst
177
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
4. Ridge preservation techniques
5. Periodontal defects
6. Closure of cleft lip and palate defects
7. Repair of oro-antral fistulas
8. Craniofacial reconstruction
II. SOFT TISSUE REGENERATION
1. Periosteal and connective tissue flaps
2. Free connective tissue and gingival
grafts
3. Root coverage procedures
4. Controlling soft tissue healing and
tissue maturity
CONTRAINDICATIONS
1. Unexplained anaemia where Hg is <12.5
g%
2. Thrombocytopenia < 100,000 / cu. mm
3. Diagnosed and treated anaemia Hg <
10.0 g%
4. Patients who have metastatic disease
5. Presence of tumour in the wound bed
6. History of platelet dysfunction
7. Active wound infection and sepsis
requiring systemic antibiotics
8. Patients with poor prognosis associated
with other disease process
9. Patient with bovine sensitivity
10. Patients with religious beliefs that
prevent the use of blood.
The resultant product consists of the
following three layers.
1. Topmost layer consists of acellular
platelet poor plasma (PPP).
2. PRF clot in the middle.
3. RBCs at the bottom.
Successful preparation of PRF requires
speedy blood collection and immediate
centrifugation, before the clotting cascade
is initiated. PRF can be obtained in the
form of a membrane by squeezing out the
fluids in the fibrin clot.
SURGICAL TECHNIQUE
The fractured root was carefully removed
and a thorough curettage of the
remaining alveolus was performed to
eliminate any residual infective tissue in
the avulsion socket that could
compromise the osseointegration of an
immediately placed implant. After
completely remove infected lesions found
that the buccal bone defect. A two - piece
implant (zimmer) of diameter 4.25 mm
and length 13 mm was placed in the
region, 21 , reaching 35 Ncm primary
stability. Then use the A-PRF to filling the
bone defect area(fig 5),and ues the APRF
membrane to cover the bone graft and
area(fig6);then suture.Fig.3-10
POST-OPERATIVE CARE
Patient was recalled after two weeks and
sutures were removed. Healing
wassatisfactory. Patient was reviewed
every two months and oral hygiene was
reinforced. Six month following the
surgery, the patient was again reviewed,
178
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
clinically the width of the ridge showed an
increase. Radiographs showed an
adequate defect fill.
The sutures were removed 10 days after
the procedure. The surgical site was
examined for uneventful healing. There
was no post - operative complications and
healing was satisfactory. The patient did
not have any post-operative morbidity.
Six months after the GBR treatment, intra-
oral examination with the bone meter
revealed adequate buccolingual width of
the ridge of 7 mm. In order not to disturb
the minimal amount of bone that would
have formed over the fenestration defect,
reentry was not performed. The implant
was uncovered and a healing abutment
was connected to allow emergence of the
implant through the soft tissues, thus
facilitating access to the implant from the
oral cavity and final restoration was
placed.Fig.11-17
DISCUSSION:
GBR is based on the principle of guided
tissue regeneration, and was first
performed in an experimental dog study.
This technique was clinically tested in the
treatment of ridge deformities.
Improvements in this technique have led
to its wide-scale clinical applications to
augment deficient alveolar ridges, treat
implant fenestration or dehiscence, and
permit immediate implant placement in
large alveolar sockets.[11,12,13]
One of the most recent developments is
the use of the autologous Platelet Rich
Fibrin [PRF] for the enhancement of the
soft and hard tissue. It was first developed
in France by Choukroun et al. It eliminates
the risk associated with the use of bovine
thrombin. PRF is an immune and platelet
concentrate which is obtained on a single
fibrin membrane, containing all the
constituents of a blood sample which are
favourable for healing and immunity.PRF
releases a number of growth factors
namely, transforming growth factor
(TGF-beta- 1), vascular endothelial growth
factor (VEGF),platelet-derived growth
factor (PDGF-AB) and thrombospondin-1
(TSP-1) during 7 days. It also secretes
fibroblast growth factor (FGF), epidermal
growth factor (EGF) and proinflammatory
cytokines like IL-1 , IL-6, and TNF- . Due to
its mechanical function and a rapid
angiogenesis promoting ability,PRF
membranes are viable material for all
types of superficial cutaneous and
mucosal healing.[3,4,7,14]
PRF in the form of a membrane can be
used in conjunction with bone grafts,
which has several advantages, such as
promoting wound healing, bone growth,
maturation and density, wound sealing
and haemostasis, and imparting better
handling properties to graft material. PRF
as an adjunct to bone graft makes it
possible to enhance the graft volume
without injuring the maturation quality in
new bone.[5,15,16]
Ιn this case-report, there was class I
fenestration defect around the implant
and to group II of the classification given
by Daniel Buser, 1994, in which the
prosthetically guided placement of an
implant results in exposure of the buccal
implant surface.[4,5,17,18]
179
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
At present, it can be stated that
biodegradable membranes have the
potential to support bone formation if
they are supported by bone graft material
to resist collapse and if they are long-
lasting enough to maintain their barrier
function for extended periods in small to
moderate bone defects.
The degradation and resorption kinetics of
a membrane for use in GBR should be set
such that it remains intact for at least 6-9
months in large volume defects and then
should be completely metabolized after
12-15months.[19,20]
In a recent systematic
review, a reasonable comparison between
bioresorbable and non-resorbable
membranes could not be drawn due to
lack of well designed studies. In this case
report, we used GTR membrane
(Healiguide), which is made of collagen
and PRF to treat the defect. This is a
significant benefit to the patient and
represents an important step in the
development of GBR procedures. For
successful outcomes with GBR, the factors
as outlined by Mellonig et al. were
followed.
Recent clinical and histologic findings
suggest that the use of platelet
concentrates have technical benefits and
may enhance bone regeneration when
used in conjunction with bone grafts.
The amplification of platelet derived
growth factor (PDGF) and transforming
growth factor (TGF) beta is seen as an
available and practical tool for enhancing
the rate of bone formation and the final
quality of bone formed.[9,11,18]
PRF has many advantages over platelet
rich plasma (PRP). It eliminates the
redundant process of adding
anticoagulant as well as the need to
neutralize it. It has been shown from the
literature that it increases the rate of
clinical graft consolidation, and PRF-
enhanced grafts produce more mature
and dense bone than do grafts without
PRF. PRF is in the form of a platelet gel
and can be used in conjunction with bone
grafts, which offers several advantages
including promoting wound healing, bone
growth and maturation, graft stabilization,
wound sealing and hemostasis and
improving the handling properties of graft
materials. In an experimental study
which used osteoblast cell cultures to
investigate the influence of PRP and PRF
on proliferation and differentiation of
osteoblasts, it was found that PRF had a
superior influence over PRP. Also, bone
augmentation grafts may act as space-
maintaining devices to allow coronal
migration of periodontal progenitor cells.
[17,18,20]
The development of biomaterials, ideally
coupled with the incorporation of bone
growth factors and bioactive peptides,
represents an important line of research
in this direction. Recent systematic
reviews regarding the survival rate of
implants into sites with
regenerated/augmented bone using
barrier membranes varied between 79%
and 100% with the majority of studies
indicating more than 90% after at least
one year of function.[4,7,8,11]
Immediate implants in the maxillary
esthetic area are currently used
frequently and are subtle, exacting
treatments. However immediate implant
180
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
placement and bone grafts are always
sensitive to the gingival quality, as the
gingival tissue has to cover and protect
the site. If the gingival tissue is weak or
damaged, dehiscence can appear in the
covering tissue leading to the
contamination of the grafted site. For this
reason, some authors recommend the use
of connective tissue grafts to reinforce the
peri-implant tissues. The L-PRF is
therefore especially indicated in this
application. The fibrin membrane of L-PRF
acts as a bio-barrier, protecting the
implant and the graft from the oral
environment. Moreover, by providing
growth factors, leukocytes, and a
permeable fibrin matrix for the growth of
endothelial and epithelial cells, this
healing material stimulates
neoangiogenesis and accelerates gingival
healing and maturation. The use of
healing materials such as L-PRF are well
suited to these applications because this
material has a robust stimulating effect on
the healing of soft and osseous tissues.
[13,17,19,20]
Thus, in this case report, a fenestration
defect was effectively treated by the
application of growth factors both to the
bone graft and GTR membrane.
CONCLUSION:
Platelet-rich fibrin (PRF) in patients with
chronic apical lesions after tooth
extraction performed immediately
implantation show high success rate of
bone graft, anti-infective ability. This
technique offers advantages for patient
comfort and the healing process. It also
facilitates a natural healing and
maturation of the periimplant bone and
soft tissues around the implant.
REFERENCES:
1. Jankovic S, Aleksic Z, Klokkevold P,
Lekovic V, Dimitrijevic B, Kenney
EB, Camargo P. Use of platelet-rich
fibrin membrane following
treatment of gingival recession: a
randomized clinical trial. Int J
Periodontics Restorative Dent
2012 Apr; 32(2):e41-50.
2. Seibert J.S. Reconstruction of
deformed, partially edentulous
ridges using full thickness onlay
grafts. I. Technique and wound
healing. Compendium of
Continuing Education in General
Dentistry; 12:437-453.
3. Choukroun. J, Diss. A, Simonpieri
A, Girard Mo, Schoeffler C, Dohan
Sl et al. Platelet Rich Fibrin (PRF), A
second generation platelet
concentrate Part I: Technological
concepts and evolution. Oral Surg
Oral Med Oral Pathol Oral Radiol
Endod 2006; 101:37-44.
4. Dohan DM, Choukroun J, Diss A,
Dohan SL, Dohan AJ, Mouhyi J, et
al. Platelet-rich fibrin (PRF): A
secondgeneration platelet
concentrate, Part I: Technological
concepts and evolution. Oral Surg
Oral Med Oral Pathol Oral Radiol
Endod 2006; 101:E37-44.
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5. Vinazzer H. Fibrin sealing:
Physiologic and
biochemicalbackground. Fac Plast
Surg 1985;2:291-295
6. Dohan DM, Choukroun J, Diss A,
Dohan SL, Dohan AJ, Mouhyi J, et
al. Platelet-rich fibrin (PRF): A
secondgeneration platelet
concentrate, Part II: Platelet-
related biologic features. Oral Surg
Oral Med Oral Pathol Oral Radiol
Endod 2006; 101:E45-50.
7. He L, Lin Y, Hu X, Zhang Y, Wu H. A
comparative study of platelet-rich
fibrin (PRF) and platelet-rich
plasma (PRP) on the effect of
proliferation and differentiation of
rat osteoblasts in vitro. Oral Surg
Oral Med Oral Pathol Oral Radiol
Endod 2009; 108:707-713.
8. Rai B ,Teoh S.H, HO ,K. H.An
evaluation of PCL-TCP composites as
delivery systems for platelet rich
plasma..Bjoms.2005;107:330
9. He L, Wu H. A comparative study of
platelet – Rich fibrin (PRF) and Platelet
rich plasma(PRP) on the effect of
proliferation and differentiation of rat
osteoblasts in vitro. Oral surg Oral
med Oral pathol Oral radiolendod
2009;108:707-13
10. Use of an Autologous Leukocyte
and Platelet-Rich Fibrin (L-PRF)
Membrane in Post-Avulsion Sites:
An Overview of Choukroun’s PRF;
The Journal of Implant & Advanced
Clinical Dentistry. 2010;56:324-327
11. Kwan Tat S, Padrines M, Theoleyre
S, Heymann D, Fortun Y.IL-6,
RANKL, TNF-alpha/IL-1:
interrelations in bone resorption
pathophysiology. Cytokine Growth
Factor Rev 2004;15: 49-60.
12. Clark RA. Fibrin and wound
healing. Ann NY Acad Sci 2001;936:
355-67.
13. Toffler M, Toscano N, Holtzclaw D,
Corso MD, Dohan Ehrenfest DM.
Introducing Choukroun’s platelet
rich fibrin (PRF) to the
reconstructive surgery milieu. The
Journal of Implant & Advanced
Clinical Dentistry 2009; 1: 21-30
14. Kfir E, Kfir V, Kaluski E. Immediate
bone augmentation after infected
tooth extraction using titanium
membranes. J Oral Implantol 2007;
33: 133-8.
15. Ortega-Martínez J, Pérez-Pascual
T, Mareque-Bueno S, Hernández-
Alfaro F, Ferrés-Padró E.
Immediate implants following
tooth extraction. A systematic
review. Med Oral Patol Oral Cir
Bucal 2012; 17:251-261
16. Gassling V, Hedderich J, Acil Y,
Purcz N, Wiltfang J, Douglas T.
Comparison of platelet rich fibrin
and collagen as osteoblast-seeded
scaffolds for bone tissue
engineering applications. Clin Oral
Implants Res 2013; 24(3):320-328.
17. Gassling V, Douglas T, Warnke PH,
Acil Y, Wiltfang J, Becker ST.
Platelet-rich fibrin membranes as
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Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
engineering. Clin Oral Implants Res
2010; 21(5):543-549.
18. Gomez-Roman G, Schulte W,
d'Hoedt B, Axman-Krcmar D. The
Frialit-2 implant system: five-year
clinical experience in single-tooth
and immediately postextraction
applications. Int J Oral Maxillofac
Implants 1997; 12(3):299-309.
19. Dohan Ehrenfest DM, Rasmusson
L, Albrektsson T. Classification of
platelet concentrates: From pure
platelet-rich plasma (P-PRP) to
leucocyte- and platelet-rich fibrin
(L-PRF). Trends Biotechnol. 2009;
27:158-167.
20. Froum SJ. Immediate placement of
implants into extraction sockets:
rationale, outcomes, technique.
Alpha Omegan 2005; 98(2):20-35.
FIGURES:
183
Figure 1: Pre op photo Figure 2: Incisal matrix made with Triad Gel to
help realign the patient's natural tooth
Figure 3. After removal of the tooth
(coronal portion)
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
Figure 4. Pilot drill in place engaging palatal
bone
Figure 5: Blue Sky Bio Bond spreader in
place
Figure 6: 3.8mm drill at 75 r.p.m.'s to
harvest some autogenous bone to fill in on
the bucca
Figure 7: autogenous bone to fill in on the
buccal
Figure 8: Zimmer Peek Provisional
abutment before preparation
Figure 9: Platelet Rich Fibrin (centrifuged
whole blood for 12 min.)
Figure 10: Patient's existing tooth (after
removal of the lingual portion) secured to
184
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
the Peek Provisional abutment in the
mouth with flowable resin
Figure 11: Closeup of the Peek abutment
cured to the patient's existing tooth
Figure 12: Closeup of the Peek abutment
cured to the patient's existing tooth
Figure 13: Bone fill on buccal with with
autogenous bone
Figure 14: Peek abutment cured to the
patient's existing tooth--screwed into
place
Figure 15: PRF membrane
Figure 16: Suture with 7/0 Prolene
sutures
185
Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858
Figure 17: Closeup
186

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EXTRACTION AND IMMEDIATE IMPLANT PLACEMENT USING A COMBINED PRF, AND PROVISIONALIZATION TECHNIQUE

  • 1. International Journal of Dental and Health Sciences Volume 01, Issue 05Case Report EXTRACTION AND IMMEDIATE IMPLANT PLACEMENT USING A COMBINED PRF, AND PROVISIONALIZATION TECHNIQUE Abu-Hussein Muhamad1 ,Abdulgani Azzaldeen2 1.University of Naples Federic II, Naples, Italy, Department of Pediatric Dentistry, University of Athens, Athens, Greece. 2. Department of Conservative Dentistry, Al-Quds University, Jerusalem, Palestine. ABSTRACT: The placement of dental implant into fresh extraction sockets was introduced in 1970 and is a well established treatment option for replacing missing teeth, allowing the restoration of masticatory function, speech and esthetics. Immediate post extraction implant placement is a well accepted protocol because of shorter total treatment time, maintenance of socket wall, reduced operative time and better actual implant placemen1 . In immediate implant placement there is gap present between implant surface and socket wall and there are various materials used to fill this gap for better osseointegration but these materials are either expensive or not so effective. Platelet- rich fibrin (PRF) is simple, natural and inexpensive. Platelet-rich fibrin(PRF) concentrate helps in healing process, osteoblastic activity, angiogenesis, release growth factors and able to stimulate defense mechanism. PRF has been used as graft material in sinus lift procedure with simultaneous implant with success. This article describes a case in which the fenestration defect around an implant was treated by the application of platelet rich fibrin, a second generation platelet concentrate along with bone graft, and guided tissue regeneration membrane. Key words: Guided bone regeneration, growth factors, platelet rich fibrin INTRODUCTION: Teeth replacement using dental implants has proven to be a successful and predictable treatment procedure; different placement and loading protocols have evolved from the first protocols in order to achieve quicker and easier surgical treatment times [1] . In situations where a tooth requires extraction and replacement, original protocol (gold standard) suggested a 6-12 month waiting period before implant placement [2] . The original protocol has been challenged within the last decade and new protocols have been developed in which implants are placed at the time of extraction of the tooth. This protocol where implants have been placed at the time of tooth *Corresponding Author Address: Abdulgani Azzaldeen,Department of Conservative Dentistry, Al-Quds University, Jerusalem, Palestine. Email: abdulganiazzaldeen@gmail.com
  • 2. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 extraction is known as immediate implants[3] . When an implant is placed in a fresh or recent extraction alveolus a gap between the implant surface and the bone walls of the socket may occur. The presence or size of the gap is both influenced by the configuration of the alveolus and by the design and width of the implant [4] . Wilson et al.[8] and Paolantonio et al.[9] demonstrated that for implants with a horizontal defect of 2 mm or less, spontaneous bone healing and osseointegration take place if the implant has a rough surface [5,6] . Horizontal defects in excess of 2 mm have been shown to not heal predictably with bone[5] . So To compensate for these problems, guided bone regeneration (GBR) using autografts, allografts, or alloplasts; barrier membranes; or combination therapy has been accomplished with what appear to be successful clinical results [3,7] . Implant therapy based on the principle of osseointegration has seen a remarkable expansion of its application in dentistry, in recent years. In the last decade, dental implants have become a reliable procedure for the treatment of partially or completely edentulous jaws. The lack of bone adjacent to an implant can be considered a true "bony defect" and several techniques have been proposed to promote defect fill with newly formed bone. One of the most popular procedures is guided bone regeneration (GBR), which involves placing a membrane over the defect to create a secluded space into which osteogenic cells can migrate and remain undisturbed over the exposed part of the implant[7,8] . Short-term animal and human studies have shown these immediate implants to be comparable to implants placed into healed bone. The advantage of the procedure include fewer surgical sessions, elimination of the waiting period for socket healing, shortened edentulous time period, reduced overall cost, as well as preservation of bone height and width [2,9] . Although immediate implantation is more demanding both surgically and prosthetically compared to the conventional placement technique, the advantages make it very appealing to patients who are in need of both extractions and implant therapy.[1,3,7,10] Platelet-rich plasma (PRP) is an autologous product that concentrates a large number of platelets in a small volume of plasma. PRP functions as a fibrin tissue adhesive with hemostatic and tissue sealing properties, but it differs from fibrin glue and other platelet-poor tissue adhesives because its platelets provide a unique ability to promote wound healing and enhance osteogenesis[1,2] . PRP provides an immediate surgical hemostatic agent that is biocompatible, safe, and effective. PRP accelerates endothelial, epithelial, and epidermal regeneration, stimulates angiogenesis, enhances collagen synthesis, promotes soft tissue healing, decreases dermal scarring, enhances the hemostatic response to injury, and reverses the inhibition of wound healing caused by 175
  • 3. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 glucocorticoids. The high leukocyte concentration of PRP has an added antimicrobial effect. Since PRP is an autologous blood product, it carries no risk of transmitting infectious disease.[4,5,6] PRP has an extremely broad range of clinical healing applications in head and neck surgery, otolaryngology, cardiovascular surgery, burns and wound healing, oral and maxillofacial surgery, cosmetic surgery, and periodontics In addition to its effectiveness for patients with chronic non-healing wounds, it has also been used as an antiangiogenic agent and as a carrier for growth factors.[1,3,4] In surgical settings, PRP decreases the frequency of intraoperative and postoperative bleeding at donor and recipient sites, accelerates soft-tissue healing, supports the initial stability of grafted tissue at recipient sites as a result of its cohesive and adhesive nature, promotes rapid vascularization of healing tissue by delivering growth factors and, when used in combination with bone replacement materials, induces regeneration.[3] The term tissue engineering was originally coined to denote the construction in the laboratory of a device containing viable cells and biologic mediators (e.g., growth factors and adhesins) in a synthetic or biologic matrix, which could be implanted in patients to facilitate regeneration of particular tissues. The role of tissue oxygenation in wound healing became the focal point in the 1980s. Tissue oxygenation enhances phagocytic and bactericidal ability of host immune cells and supports collagen as well as other protein synthetic events. The importance of growth factors in enhancing wound healing has become the focus of research in the present day. In addition, a link has been established between tissue oxygenation and growth factors. Macrophage stimulation causes the release of angiogenic and other growth factors that support wound healing and resist infection. In general, tissue engineering combines three key elements, namely scaffolds (collagen, bone mineral), signaling molecules (growth factors), and cells (osteoblasts, fibroblasts). Tissue engineering has been redefined presently as the relatively new, highly promising field of reconstructive biology, which draws on the recent advances in medicine and surgery, molecular and cellular biology, polymer chemistry, and physiology.[4,5,6] Platelets isolated from peripheral blood are an autologous source of growth factors. When platelets in a concentrated form are added to graft materials, a more predictable outcome is derived. Platelet- rich plasma (PRP) is an easily accessible source of growth factors to support bone- and soft-tissue healing. It is derived by methods that concentrate autologous platelets and is added to surgical wounds or grafts and to other injuries in need of supported or accelerated healing. A blood clot is the center focus of initiating any soft-tissue healing and bone regeneration. In all natural wounds, a blood clot forms and starts the healing process. PRP is a simple strategy to concentrate platelets or enrich natural blood clot, which forms in 176
  • 4. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 normal surgical wounds, to initiate a more rapid and complete healing process. A natural blood clot contains 95% red blood cells, 5% platelets, less than 1% white blood cells, and numerous amounts of fibrin strands. A PRP blood clot contains 4% red blood cells, 95% platelets, and 1% white blood cells[2,7,9] . The use of PRP in place of recombinant growth factors has several advantages, in that growth factors obtained from platelets not only have their own specific action on tissues but also interact with other growth factors, resulting in the activation of gene expression and protein production. Therefore, the properties of PRP are based on the production and release of multiple growth and differentiation factors upon platelet activation. These factors are critical in the regulation and stimulation of the wound healing process, and they play an important role in regulating cellular processes such as mitogenesis, chemotaxis, differentiation, and metabolism. Platelet rich fibrin, which is a second generation platelet concentrate, offers the surgeon an access to growth factors with a simple and available technology. These growth factors which are autologous, nontoxic and non immunogenic, enhance and accelerate the normal bone regeneration pathways. [10,11,12] This article presents a case report in which a fenestration defect around an implant was treated by GBR with platelet rich fibrin (PRF)-enhanced bone graft and the PRF smeared barrier membrane for guided bone regeneration. CASE DETAIL: A healthy 27-year-old male oral surgery resident was referred to the author’s private office for a apical lesions in the maxillary anterior teeth need to be removed and implant restoration . He gave the history of trauma four months back and simultaneous avulsion of the left central incisor tooth. On clinical examination , height of the alveolar ridge was adequate but the width of the ridge was inadequate for an implant placement. The ridge was found to be a class I defect according to Siebert’s classification. Intraoral radiographs also revealed a deficient ridge. Ridge augmentation using bone graft with platelet rich fibrin (PRF) was planned. The patient was clinically healthy with no history of systemic diseases. Informed consent was taken from the patient.Fig.1-2 PREPARATION OF PRF: PRF belongs to a new generation of platelet concentrate but with a simplified processing as compared to platelet rich plasma (PRP). Blood was drawn into 10ml test tubes without an anticoagulant and centrifuged immediately for 12 minutes at 2,700 rpm . Indication for using PRF I. BONE REGENERATION : 1. Sinus lift grafting 2. Ridge augmentation 3. Repair of bone defects created by removal of teeth or small cyst 177
  • 5. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 4. Ridge preservation techniques 5. Periodontal defects 6. Closure of cleft lip and palate defects 7. Repair of oro-antral fistulas 8. Craniofacial reconstruction II. SOFT TISSUE REGENERATION 1. Periosteal and connective tissue flaps 2. Free connective tissue and gingival grafts 3. Root coverage procedures 4. Controlling soft tissue healing and tissue maturity CONTRAINDICATIONS 1. Unexplained anaemia where Hg is <12.5 g% 2. Thrombocytopenia < 100,000 / cu. mm 3. Diagnosed and treated anaemia Hg < 10.0 g% 4. Patients who have metastatic disease 5. Presence of tumour in the wound bed 6. History of platelet dysfunction 7. Active wound infection and sepsis requiring systemic antibiotics 8. Patients with poor prognosis associated with other disease process 9. Patient with bovine sensitivity 10. Patients with religious beliefs that prevent the use of blood. The resultant product consists of the following three layers. 1. Topmost layer consists of acellular platelet poor plasma (PPP). 2. PRF clot in the middle. 3. RBCs at the bottom. Successful preparation of PRF requires speedy blood collection and immediate centrifugation, before the clotting cascade is initiated. PRF can be obtained in the form of a membrane by squeezing out the fluids in the fibrin clot. SURGICAL TECHNIQUE The fractured root was carefully removed and a thorough curettage of the remaining alveolus was performed to eliminate any residual infective tissue in the avulsion socket that could compromise the osseointegration of an immediately placed implant. After completely remove infected lesions found that the buccal bone defect. A two - piece implant (zimmer) of diameter 4.25 mm and length 13 mm was placed in the region, 21 , reaching 35 Ncm primary stability. Then use the A-PRF to filling the bone defect area(fig 5),and ues the APRF membrane to cover the bone graft and area(fig6);then suture.Fig.3-10 POST-OPERATIVE CARE Patient was recalled after two weeks and sutures were removed. Healing wassatisfactory. Patient was reviewed every two months and oral hygiene was reinforced. Six month following the surgery, the patient was again reviewed, 178
  • 6. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 clinically the width of the ridge showed an increase. Radiographs showed an adequate defect fill. The sutures were removed 10 days after the procedure. The surgical site was examined for uneventful healing. There was no post - operative complications and healing was satisfactory. The patient did not have any post-operative morbidity. Six months after the GBR treatment, intra- oral examination with the bone meter revealed adequate buccolingual width of the ridge of 7 mm. In order not to disturb the minimal amount of bone that would have formed over the fenestration defect, reentry was not performed. The implant was uncovered and a healing abutment was connected to allow emergence of the implant through the soft tissues, thus facilitating access to the implant from the oral cavity and final restoration was placed.Fig.11-17 DISCUSSION: GBR is based on the principle of guided tissue regeneration, and was first performed in an experimental dog study. This technique was clinically tested in the treatment of ridge deformities. Improvements in this technique have led to its wide-scale clinical applications to augment deficient alveolar ridges, treat implant fenestration or dehiscence, and permit immediate implant placement in large alveolar sockets.[11,12,13] One of the most recent developments is the use of the autologous Platelet Rich Fibrin [PRF] for the enhancement of the soft and hard tissue. It was first developed in France by Choukroun et al. It eliminates the risk associated with the use of bovine thrombin. PRF is an immune and platelet concentrate which is obtained on a single fibrin membrane, containing all the constituents of a blood sample which are favourable for healing and immunity.PRF releases a number of growth factors namely, transforming growth factor (TGF-beta- 1), vascular endothelial growth factor (VEGF),platelet-derived growth factor (PDGF-AB) and thrombospondin-1 (TSP-1) during 7 days. It also secretes fibroblast growth factor (FGF), epidermal growth factor (EGF) and proinflammatory cytokines like IL-1 , IL-6, and TNF- . Due to its mechanical function and a rapid angiogenesis promoting ability,PRF membranes are viable material for all types of superficial cutaneous and mucosal healing.[3,4,7,14] PRF in the form of a membrane can be used in conjunction with bone grafts, which has several advantages, such as promoting wound healing, bone growth, maturation and density, wound sealing and haemostasis, and imparting better handling properties to graft material. PRF as an adjunct to bone graft makes it possible to enhance the graft volume without injuring the maturation quality in new bone.[5,15,16] Ιn this case-report, there was class I fenestration defect around the implant and to group II of the classification given by Daniel Buser, 1994, in which the prosthetically guided placement of an implant results in exposure of the buccal implant surface.[4,5,17,18] 179
  • 7. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 At present, it can be stated that biodegradable membranes have the potential to support bone formation if they are supported by bone graft material to resist collapse and if they are long- lasting enough to maintain their barrier function for extended periods in small to moderate bone defects. The degradation and resorption kinetics of a membrane for use in GBR should be set such that it remains intact for at least 6-9 months in large volume defects and then should be completely metabolized after 12-15months.[19,20] In a recent systematic review, a reasonable comparison between bioresorbable and non-resorbable membranes could not be drawn due to lack of well designed studies. In this case report, we used GTR membrane (Healiguide), which is made of collagen and PRF to treat the defect. This is a significant benefit to the patient and represents an important step in the development of GBR procedures. For successful outcomes with GBR, the factors as outlined by Mellonig et al. were followed. Recent clinical and histologic findings suggest that the use of platelet concentrates have technical benefits and may enhance bone regeneration when used in conjunction with bone grafts. The amplification of platelet derived growth factor (PDGF) and transforming growth factor (TGF) beta is seen as an available and practical tool for enhancing the rate of bone formation and the final quality of bone formed.[9,11,18] PRF has many advantages over platelet rich plasma (PRP). It eliminates the redundant process of adding anticoagulant as well as the need to neutralize it. It has been shown from the literature that it increases the rate of clinical graft consolidation, and PRF- enhanced grafts produce more mature and dense bone than do grafts without PRF. PRF is in the form of a platelet gel and can be used in conjunction with bone grafts, which offers several advantages including promoting wound healing, bone growth and maturation, graft stabilization, wound sealing and hemostasis and improving the handling properties of graft materials. In an experimental study which used osteoblast cell cultures to investigate the influence of PRP and PRF on proliferation and differentiation of osteoblasts, it was found that PRF had a superior influence over PRP. Also, bone augmentation grafts may act as space- maintaining devices to allow coronal migration of periodontal progenitor cells. [17,18,20] The development of biomaterials, ideally coupled with the incorporation of bone growth factors and bioactive peptides, represents an important line of research in this direction. Recent systematic reviews regarding the survival rate of implants into sites with regenerated/augmented bone using barrier membranes varied between 79% and 100% with the majority of studies indicating more than 90% after at least one year of function.[4,7,8,11] Immediate implants in the maxillary esthetic area are currently used frequently and are subtle, exacting treatments. However immediate implant 180
  • 8. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 placement and bone grafts are always sensitive to the gingival quality, as the gingival tissue has to cover and protect the site. If the gingival tissue is weak or damaged, dehiscence can appear in the covering tissue leading to the contamination of the grafted site. For this reason, some authors recommend the use of connective tissue grafts to reinforce the peri-implant tissues. The L-PRF is therefore especially indicated in this application. The fibrin membrane of L-PRF acts as a bio-barrier, protecting the implant and the graft from the oral environment. Moreover, by providing growth factors, leukocytes, and a permeable fibrin matrix for the growth of endothelial and epithelial cells, this healing material stimulates neoangiogenesis and accelerates gingival healing and maturation. The use of healing materials such as L-PRF are well suited to these applications because this material has a robust stimulating effect on the healing of soft and osseous tissues. [13,17,19,20] Thus, in this case report, a fenestration defect was effectively treated by the application of growth factors both to the bone graft and GTR membrane. CONCLUSION: Platelet-rich fibrin (PRF) in patients with chronic apical lesions after tooth extraction performed immediately implantation show high success rate of bone graft, anti-infective ability. This technique offers advantages for patient comfort and the healing process. It also facilitates a natural healing and maturation of the periimplant bone and soft tissues around the implant. REFERENCES: 1. Jankovic S, Aleksic Z, Klokkevold P, Lekovic V, Dimitrijevic B, Kenney EB, Camargo P. Use of platelet-rich fibrin membrane following treatment of gingival recession: a randomized clinical trial. Int J Periodontics Restorative Dent 2012 Apr; 32(2):e41-50. 2. Seibert J.S. Reconstruction of deformed, partially edentulous ridges using full thickness onlay grafts. I. Technique and wound healing. Compendium of Continuing Education in General Dentistry; 12:437-453. 3. Choukroun. J, Diss. A, Simonpieri A, Girard Mo, Schoeffler C, Dohan Sl et al. Platelet Rich Fibrin (PRF), A second generation platelet concentrate Part I: Technological concepts and evolution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006; 101:37-44. 4. Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJ, Mouhyi J, et al. Platelet-rich fibrin (PRF): A secondgeneration platelet concentrate, Part I: Technological concepts and evolution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006; 101:E37-44. 181
  • 9. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 5. Vinazzer H. Fibrin sealing: Physiologic and biochemicalbackground. Fac Plast Surg 1985;2:291-295 6. Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJ, Mouhyi J, et al. Platelet-rich fibrin (PRF): A secondgeneration platelet concentrate, Part II: Platelet- related biologic features. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006; 101:E45-50. 7. He L, Lin Y, Hu X, Zhang Y, Wu H. A comparative study of platelet-rich fibrin (PRF) and platelet-rich plasma (PRP) on the effect of proliferation and differentiation of rat osteoblasts in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 108:707-713. 8. Rai B ,Teoh S.H, HO ,K. H.An evaluation of PCL-TCP composites as delivery systems for platelet rich plasma..Bjoms.2005;107:330 9. He L, Wu H. A comparative study of platelet – Rich fibrin (PRF) and Platelet rich plasma(PRP) on the effect of proliferation and differentiation of rat osteoblasts in vitro. Oral surg Oral med Oral pathol Oral radiolendod 2009;108:707-13 10. Use of an Autologous Leukocyte and Platelet-Rich Fibrin (L-PRF) Membrane in Post-Avulsion Sites: An Overview of Choukroun’s PRF; The Journal of Implant & Advanced Clinical Dentistry. 2010;56:324-327 11. Kwan Tat S, Padrines M, Theoleyre S, Heymann D, Fortun Y.IL-6, RANKL, TNF-alpha/IL-1: interrelations in bone resorption pathophysiology. Cytokine Growth Factor Rev 2004;15: 49-60. 12. Clark RA. Fibrin and wound healing. Ann NY Acad Sci 2001;936: 355-67. 13. Toffler M, Toscano N, Holtzclaw D, Corso MD, Dohan Ehrenfest DM. Introducing Choukroun’s platelet rich fibrin (PRF) to the reconstructive surgery milieu. The Journal of Implant & Advanced Clinical Dentistry 2009; 1: 21-30 14. Kfir E, Kfir V, Kaluski E. Immediate bone augmentation after infected tooth extraction using titanium membranes. J Oral Implantol 2007; 33: 133-8. 15. Ortega-Martínez J, Pérez-Pascual T, Mareque-Bueno S, Hernández- Alfaro F, Ferrés-Padró E. Immediate implants following tooth extraction. A systematic review. Med Oral Patol Oral Cir Bucal 2012; 17:251-261 16. Gassling V, Hedderich J, Acil Y, Purcz N, Wiltfang J, Douglas T. Comparison of platelet rich fibrin and collagen as osteoblast-seeded scaffolds for bone tissue engineering applications. Clin Oral Implants Res 2013; 24(3):320-328. 17. Gassling V, Douglas T, Warnke PH, Acil Y, Wiltfang J, Becker ST. Platelet-rich fibrin membranes as scaffolds for periosteal tissue 182
  • 10. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 engineering. Clin Oral Implants Res 2010; 21(5):543-549. 18. Gomez-Roman G, Schulte W, d'Hoedt B, Axman-Krcmar D. The Frialit-2 implant system: five-year clinical experience in single-tooth and immediately postextraction applications. Int J Oral Maxillofac Implants 1997; 12(3):299-309. 19. Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Classification of platelet concentrates: From pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol. 2009; 27:158-167. 20. Froum SJ. Immediate placement of implants into extraction sockets: rationale, outcomes, technique. Alpha Omegan 2005; 98(2):20-35. FIGURES: 183 Figure 1: Pre op photo Figure 2: Incisal matrix made with Triad Gel to help realign the patient's natural tooth Figure 3. After removal of the tooth (coronal portion)
  • 11. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 Figure 4. Pilot drill in place engaging palatal bone Figure 5: Blue Sky Bio Bond spreader in place Figure 6: 3.8mm drill at 75 r.p.m.'s to harvest some autogenous bone to fill in on the bucca Figure 7: autogenous bone to fill in on the buccal Figure 8: Zimmer Peek Provisional abutment before preparation Figure 9: Platelet Rich Fibrin (centrifuged whole blood for 12 min.) Figure 10: Patient's existing tooth (after removal of the lingual portion) secured to 184
  • 12. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 the Peek Provisional abutment in the mouth with flowable resin Figure 11: Closeup of the Peek abutment cured to the patient's existing tooth Figure 12: Closeup of the Peek abutment cured to the patient's existing tooth Figure 13: Bone fill on buccal with with autogenous bone Figure 14: Peek abutment cured to the patient's existing tooth--screwed into place Figure 15: PRF membrane Figure 16: Suture with 7/0 Prolene sutures 185
  • 13. Muhamad A. et al., Int J Dent Health Sci 2014; 1(5): 847-858 Figure 17: Closeup 186