A combination graft of low-molecular-weight silk fibroin with

Choukroun platelet-rich fibrin for rabbit calvarial defect

Eui-Hee Lee, DDS,a Jwa-Young Kim, DDS,b Hae Yong Kweon, PhD,c You-Young Jo, PhD,d
Soo-Kee Min, MD, PhD,e Young-Wook Park, DDS, PhD,f Je-Yong Choi, DDS, PhD,g and
Seong-Gon Kim, DDS, PhD,h Kyoungkido, Daegu, and Gangneung, Korea
HALLYM UNIVERSITY, RURAL DEVELOPMENT ADMINISTRATION, KYUNGPOOK NATIONAL
UNIVERSITY, AND GANGNEUNG-WONJU NATIONAL UNIVERSITY

Objective. The objective of this study was to determine the capabilities of silk fibroin as a biomaterial template for
bone formation when mixed with Choukroun platelet-rich fibrin (PRF) in vivo.
Study design. Ten New Zealand white rabbits were used for this study and bilateral round shaped defects were
formed in the parietal bone (diameter 9.0 mm). The silk fibroin was digested by acid and made into powder
(molecular weight ⬍1.0 kDa). The right side (experimental group) received the silk fibroin plus platelet-rich fibroin and
the left side (control group) did not receive a graft. Animals were killed at 6 weeks and 12 weeks. The specimens were
examined by microscopic computerized tomography (␮-CT). Subsequently, they underwent decalcification and were
stained for histologic analysis.
Results. There was no significant difference between groups at 6 weeks after operation. In the ␮-CT results, however,
tissue mineral content in the experimental group at 12 weeks after operation was 132.09 ⫾ 4.41 and that in the
control group was 126.42 ⫾ 6.62 (P ⫽ .011). Tissue mineral density in the experimental group was 2,088.88 ⫾
648.34, and that in the control group was 2,029.72 ⫾ 668.22 (P ⫽ .013). The results of the histomorphometric
analysis were in accordance with the ␮-CT results. The total new bone was 49.86 ⫾ 7.49% in the control group at 12
weeks after the operation and 59.83 ⫾ 10.92% in the experimental group (P ⫽ .021).
Conclusion. A combined application of Choukroun PRF with acid-digested silk fibroin showed more rapid bone
healing than unfilled control. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;109:e33-e38)

Bony defects can result from pathologic lesions or
surgical procedures. For the reconstruction of bony
defects, autogenous bone graft is the treatment of
choice. However, it has many limitations, such as its
limited availability, donor site morbidity, and the pro-
Supported by the BioGreen21 Program (grant nos. 200810FTH010103002
and 200810FTH010102001), Rural Development Administration.
a
Resident, Department of Oral and Maxillofacial Surgery, Sacred
Heart Hospital, Hallym University.
b
Instructor, Department of Oral and Maxillofacial Surgery, Sacred
Heart Hospital, Hallym University.
c
Senior Researcher, National Academy of Agricultural Science, Rural
Development Administration.
d
Senior Researcher, National Academy of Agricultural Science, Ru-
ral Development Administration.
e
Associate Professor, Department of Pathology, Sacred Heart Hospi-
tal, Hallym University.
f
Professor, Department of Oral and Maxillofacial Surgery, Gan-
gneung-Wonju National University.
g
Professor, Department of Biochemistry and Cell Biology, Kyung-
pook National University.
h
Assistant Professor, Department of Oral and Maxillofacial Surgery,
Gangneung-Wonju National University.
Received for publication Aug 18, 2009; returned for revision Dec 9,
2009; accepted for publication Dec 28, 2009.
1079-2104/$ - see front matter
© 2010 Mosby, Inc. All rights reserved.
doi:10.1016/j.tripleo.2009.12.043
longed time of operation. Material originating from
autogenous blood can be used as filling material for the
bony defect instead of bone, in which case donor site
morbidity and operating time can be reduced. Platelet-
rich plasma (PRP) has been widely used for this pur-
pose. It can be used alone or in combination with other
biomaterials.1 However, the preparation of PRP re-
quires several steps and chemical additives.1 Chouk-
roun platelet-rich fibrin (PRF) is a modification of PRP
and can be prepared in a single step without any addi-
tive.2
Fibrin is derived from a form of plasmatic molecule
called fibrinogen. The fibrillary molecule is present in
plasma and plays an important role in platelet aggrega-
tion during hemostasis.3 Thrombin acts on fibrinogen to
form 1 molecule of fibrin monomer that has the ability
to polymerize with other fibrin monomer molecules to
form fibrin fiber.4 The addition of fibrin sealant with
resorptive ceramic biomaterial increases bone forma-
tion in the bony defects of rabbits’ tibia.5 Fibrin adhe-
sive can be used to maintain bone graft fragments in a
mass.6 However, the fibrin without bone substitute ma-
terial cannot increase bone regeneration and even in-
hibits normal healing.7 This fibrin can inhibit bony
integration into some biomaterials, such as Bio-Oss.8

e33

e34 Lee et al.
Fig. 1. A, Scanning electron microscopic image of silk powder. Because the silk fibroin macromolecule was randomly degraded
by acid, the size of each particle was different. B, The blood was sampled from the ear of the rabbit. C, After centrifugation, the
blood was separated in 3 layers (arrows). D, A combination of silk powder and platelet-rich fibroin was placed on the right side
(asterisk), and the left side was left unfilled.
Therefore, caution must be exercised when selecting
the biomaterial to combine with fibrin.
Silk is composed of protein polymers that are produced
by spiders9 and larvae such as silkworms.10 Silk protein
has a repetitive protein sequence consisting of 4 different
structural components that lead to homogeneity in the
secondary structure.11 Silk fibroin has been used in tissue
engineering as a scaffold for the repair of ligament, ten-
don, bone, and cartilage. Although silk may induce in-
flammatory and immunogenic reactions, it can safely be
used when it is prepared without glycosylated protein.12
As a scaffold, it delivers inductive cells to the healing site
and provides cues to control the structure of newly formed
tissue.13 Silk fibroin– based scaffold shows good biocom-
patibility,14 slow degradation,15 and excellent mechanical
properties.16 Silk is insoluble in organic solvent.15 Be-
cause silk fibroin is an excellent scaffold, silk fibroin may
be used as a scaffold for Choukroun PRF.
The objective of the present study was to determine the
capability of silk fibroin as a biomaterial template for bone
formation when mixed with Choukroun PRF in vivo.
MATERIAL AND METHOD
Animals and materials
Ten 3-month-old New Zealand white rabbits with an
average weight of 2.3 kg (range 2.0-2.5 kg) were used in
this experiment. This experiment was approved by the
OOOOE
May 2010
Institutional Animal Care and Use Committee of the Bio-
venture Incubation Center, Hanbat National University,
Daejeon, Korea (no. 2009-NCT-004). The low-molec-
ular-weight silk fibroin powder (molecular weight 0.5-
1.0 kDa) was prepared by the Rural Development Ad-
ministration (Suwon, Korea) and kindly donated for
this experiment (Fig. 1, A). Briefly, silk fibroin macro-
molecules without sericin were dissolved in hydrochlo-
ric acid, and an electrodialysis system was used to
remove salt. Then it was made into a powder.
Surgical method
General anesthesia was induced by intramuscular
injection of a combination of 0.4 mL ketamine (100
mg/mL; Ketara; Yuhan, Seoul, Korea) and 0.3 mL
xylazine (10 mg/kg body weight; Rompun; Bayer Ko-
rea, Seoul, Korea). Additionally, a 10-mL blood sample
was obtained through the ear vein of each rabbit (Fig. 1,
B). The sample was then centrifuged for 12 minutes at
a rate of 400g. After centrifugation, the blood was
separated into 3 layers (Fig. 1, C). Among these, the
middle layer, representing Choukroun PRF, was taken.
The cranium area was shaved and disinfected with
povidine-iodine. From the nasal bone to the occipital
protuberance, a longitudinal incision was made in the
skull. Then a midline incision was created in the peri-
osteum. Sharp subperiosteal dissection reflected the
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Volume 109, Number 5
Table I. Microscopic computerized tomography analysis
6 weeks
Unfilled Silk fibroin ⫹ PRF
BMC (mg) 159.57 ⫾ 25.50 181.01 ⫾ 3.91
BMD (mg/mm3) 1,059.66 ⫾ 166.69 1,202.46 ⫾ 28.99
TMC 60.19 ⫾ 20.26 72.80 ⫾ 3.40
TMD 1,867.98 ⫾ 147.12 1,989.24 ⫾ 117.30
PRF, Platelet-rich-fibroin; BMC, bone mineral content; BMD, bone mineral density; TMC, tissue mineral content; TMD, tissue mineral density;
NS, not significant.
pericranium from the outer table of the cranial valut,
exposing the parietal bones. A dental-trephine bur was
used under copious saline irrigation to create a bilateral
full-thickness calvarial defect. Two 9-mm-diameter de-
fects were created, one on each side of the midline. The
silk powder mixed with Choukroun PRF was placed on
the right side, and the left side was left empty (Fig. 1,
D). The experimental designs of previously published
studies were referenced in devising the present exper-
iment.17,18 Then the pericranium and skin were closed
in layers with 3-0 silk. After surgery, the rabbits re-
ceived gentamicin 1 mg/kg (Kookje, Seoul, Korea)
intramuscularly 3 times daily for 3 days. Each rabbit
was individually caged and received food and water.
Previously published studies were referenced to deter-
mine the observation point.19,20 Five animals were
killed at 6 weeks and 5 at 12 weeks. The dimension of
the calvarial specimens was 25 ⫻ 12 ⫻ 3 mm at the
largest including both defects. They were fixed in 10%
formalin and underwent microscopic computerized to-
mography (␮-CT).
Microscopic computerized tomography
The prepared specimens underwent ␮-CT using an
Explored Locus SP ␮-CT scanner (GE Medical Sys-
tems, London, Canada). After the calibration, the cal-
varial specimens were scanned in sections of 0.05 mm
thickness. The scanned images were reconstructed by
Microview software (GE Medical Systems). The cali-
brated 3-dimensional images were shown in the gross
profiles of the specimens. Because the initial defect was
round in shape with 9.0 mm diameter, the setting of the
region of interest (ROI) was considered to be the initial
defect size and shape. A threshold level of 25% of the
bone standard was set as recommended by the manu-
facturer.
The ROI of each specimen was analyzed for bone
mineral content (BMC) and bone mineral density
(BMD). The tissue mineral content (TMC) and tissue
mineral density (TMD) were calculated using appropri-
ate software.
Lee et al. e35
12 weeks
P value Unfilled Silk fibroin ⫹ PRF P value
NS 271.25 ⫾ 20.69 279.37 ⫾ 14.82 NS
NS 1,467.61 ⫾ 480.28 1,550.46 ⫾ 415.06 NS
NS 126.42 ⫾ 6.62 132.09 ⫾ 4.41 .011
NS 2,029.72 ⫾ 668.22 2,088.88 ⫾ 648.34 .013
Histomorphometric evaluation
After radiodensitometry analysis, the calvaria were
subjected to dehydration and embedding. The bones
were dehydrated in ethanol and decalcified using 5%
formic acid for 2 weeks. The right and left parietal
bones were separated through the midline sagittal su-
ture. Both segments were embedded to show the sag-
ittal sections in paraffin blocks. Then the sections were
sliced and stained with hematoxylin and eosin. The
section showing the widest defect area was selected and
the cuts from 50 ␮im before and after were also se-
lected. Digital images of selected sections were taken
using a digital camera (DP-20; Olympus, Tokyo, Ja-
pan). The images were analyzed by Sigma Scan Pro
(SPSS, Chicago, IL). Total new bone was calculated as
a percentage of the total region of the defect.
Statistical analysis
A paired t test was used for comparison of the
samples from the same animal. The statistically signif-
icant level was set as P ⬍ .05.
RESULTS
Microscopic computerized tomography
The results of the ␮-CT analysis are shown in Table
I. The average value of all measured variables was
higher in the experimental group than in the control
group at 6 weeks after the operation (Fig. 2, A). How-
ever, there was no statistically significant difference
(P ⬎ .05). The size of the remaining defect was larger
in the control group than in the experimental group
(Fig. 2, B). The TMC in the experimental group at 12
weeks after operation was 132.09 ⫾ 4.41 and that in the
control group was 126.42 ⫾ 6.62 (Table I). This was a
statistically significant difference (P ⫽ .011). The TMD
in the experimental group at 12 weeks after operation
was 2,088.88 ⫾ 648.34 and that in the control group
was 2,029.72 ⫾ 668.22 (Table I). This was a statisti-
cally significant difference (P ⫽ .013). The other vari-
ables failed to show a statistically significant difference
(P ⬎ .05).
 OOOOE
e36 Lee et al. May 2010

Fig. 2. Microscopic computerized tomography. A, The defect size was smaller in the experimental group (asterisk) than in the
control group at 6 weeks. B, Most bony defects were filled by new bone at 12 weeks. The relative size of the defects was small
in the experimental group (asterisk).

Table II. Histomorphometric analysis

6 weeks 12 weeks
Unfilled Silk fibroin ⫹ PRF P value Unfilled Silk fibroin ⫹ PRF P value
Total new bone (%) 36.59 ⫾ 6.11 44.38 ⫾ 17.00 NS 49.86 ⫾ 7.49 59.83 ⫾ 10.92 .021
PRF, Platelet-rich-fibroin; NS, not significant.

Fig. 3. Histologic view at 6 weeks. The size of the unfilled defect was larger in the control group (A) than in the experimental
group (B). C, Magnified image of the control group (original magnification ⫻200). D, The experimental groups showed new bone
formation in the middle of the defect (original magnification ⫻200). Bar length ⫽ 2 mm.

Histomorphometry at 12 weeks after the operation (Fig. 4, B and D). The

The results of histomorphometry are shown in Table
II. The total new bone was 36.59 ⫾ 6.11% in the
control group at 6 weeks after the operation (Fig. 3, A
and C). It was 44.38 ⫾ 17.00% in the experimental
group at 6 weeks after the operation (Fig. 3, B and D).
However, the difference was not statistically significant
(P ⬎ .05). The total new bone was 49.86 ⫾ 7.49% in the
control group at 12 weeks after the operation (Fig. 4, A
and C). It was 59.83 ⫾ 10.92% in the experimental group
difference was statistically significant (P ⫽ .021).
DISCUSSION
In this study, silk fibroin powder was used as a
combination template with Choukroun PRF for the
restoration of bony defect. Since fibrin without bone
substitute material cannot increase bone regeneration
and even inhibits normal healing,7 the silk powder can
be considered a scaffold for Choukroun PRF.
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Volume 109, Number 5
Fig. 4. Histologic view at 12 weeks. The size of the unfilled defect was larger in the control group (A) than in the experimental
group (B). C, Magnified image of the control group showed dense fibrotic tissue (original magnification ⫻200). D, The
experimental groups showed narrow defects filled with loose fiber (original magnification ⫻200). Bar length ⫽ 2 mm.
Silk fibroin is produced by Bombyx mori and has
been widely studied as a tissue scaffold.12-14 The ad-
vantages of the silk fibroin are that it is less expensive
and has good biocompatibility, slow degradation, and
plasticity.21,22 These advantages are highly important
for the regeneration of connective tissue. However,
some features, such as slow degradation, may have an
adverse effect on bone regeneration. It is also known to
induce inflammatory reaction.23,24 For the successful
regeneration of bone, degradation velocity and inflam-
mation should be controlled.
The silk structure is modified in an attempt to over-
come the limitations of silk as a bone substitute. When
silk fibroin film is chemically mixed with RGD peptide,
bone formation in vitro is significantly improved.25,26
The addition of bone morphogenetic protein 2 and
nanohydroxyapatite to the silk fibroin scaffold can sig-
nificantly increase bone formation.13 Thin silk fibroin
film scaffolds with stem cells presented new bone for-
mation.27 Although silk fibroin can induce the activity
of alkaline phosphatase, the silk fibroin does not have
enough osteogenic power to induce new bone without
any modification.28
Choukroun PRF was produced from the patient.
Therefore, there were no immunologic or infection-
related concerns. The amount of Choukroun PRF may
have been limited because of the limited sampling size.
When the defect size was relatively large, Choukroun
PRF had to be used with other filling materials. Ac-
cording to the ␮-CT results, silk fibroin powder was
used as a combination template with Choukroun PRF
(Table I). It was revealed that the TMC in the experi-
mental group was significantly higher than that of the
Lee et al. e37
control group (P ⫽ .011; Table I). The TMD was also
significantly higher in the experimental group than in
the control group (P ⫽ .013; Table I). The results were
further confirmed by histomorphometric analysis (Ta-
ble II). The total new bone was 49.86 ⫾ 7.49% in the
control group at 12 weeks after the operation and
59.83 ⫾ 10.92% in the experimental group (P ⫽ .021).
The ␮-CT analysis was highly correlated to the histo-
morphometric analysis.29 Our study also showed a
close correlation between the two analyses.
Silk fibroin is a macromolecule and can be degraded
by enzymes in vivo.30 Though the degradation time of
silk is reported differently, it takes 2 years for the
complete loss of tensile strength.31 During the degra-
dation, silk induces an immunogenic reaction mediated
by lymphocytes and multinucleated giant cells.23,24
Biodegradation is the breakdown of macromolecules
into small fragments.30 Because silk fibroin fragments
were used in the present study, the presence of an
immunogenic reaction was hardly identified in the his-
tologic exam. The silk fibroin fragment seemed to be
degraded completely within 6 weeks. The inflammatory
reaction was minimal, unlike for the silk fibroin mac-
romolecule. Our silk powder increased the alkaline
phosphatase activity in proportion to the applied dose in
the osteoblast-like cells (data not shown). Therefore, in
terms of inflammatory potential and osteoinduction, our
silk powder was more appropriate as a scaffold for bone
regeneration than the silk macromolecule was. Because
the presented product was a powder, its application
might be limited to cavity-form deformities. Ideal bio-
degradability as a scaffold for bone regeneration is
largely unknown. Degradation that is too rapid may not

e38 Lee et al.
induce new bone formation, whereas degradation that is
too slow may prevent the space occupied by the graft
from being changed to bone. The biodegradability of
silk fibroin might be controlled by the length of the silk
fibroin fragment.30 However, the relationship between
biodegradability and osteoinduction is still unclear.
Most of the osteogenic fragments of the digested silk
fibroin will be studied in future research.
In conclusion, a combined application of Choukroun
PRF with the acid-digested silk fibroin showed more
rapid bone healing than the unfilled control. Because
silk is a cheap and readily available material, silk
fibroin and Choukroun PRF combination therapy could
provide a possible new bone substitute for the recon-
struction of various bony defects. However, results
from these studies on rabbits cannot be directly extrap-
olated to human application, so further study is needed.
REFERENCES
1. Kim SG, Kim WK, Park JC, Kim HJ. A comparative study of
osseointegration of Avana implants in a demineralized freeze-
dried bone alone or with platelet-rich plasma. J Oral Maxillofac
Surg 2002;60:1018-25.
2. Soffer E, Ouhayoun JP, Anagnostou F. Fibrin sealants and plate-
let preparations in bone and periodontal healing. Oral Surg Oral
Med Oral Pathol Oral Radio Endod 1998;85:638-46.
3. Dohan MD, Choukroun J, Diss A, Dohan SL, Dohan AJ, Mouhyi
J, 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:e37-44.
4. Mannaioni PF, Di Bello MG, Masini E. Platelets and inflamma-
tion: role of platelet-derived growth factor, adhesion molecules
and histamine. Inflamm Res 1997;46:4-18.
5. Kania RE, Meunier A, Hamadouche M, Sedel L, Petite H.
Addition of fibrin sealant to ceramic promotes bone repair:
long-term study in rabbit femoral defect model. J Biomed Mater
Res 1998;43:38-45.
6. Tayapongsak P, O’Brien DA, Monteiro CB, Arceo-Diaz LY.
Autologus fibrin adhesive in mandibular reconstruction with
particulate cancellous bone and marrow. J Oral Maxillofac Sug
1994;52:161-5.
7. Jung RE, Schmoekel HG. Platelet-rich plasma and fibrin as delivery
system for rhBMP-2. Clin Oral Implants Res 2005;16:676-82.
8. Carmagnola D, Berglundh T, Lindhe J. The effect of a fibrin glue
on the integration of Bio-Oss with bone tissue. An experimental
study in labrador dogs. J Clin Periodontol 2002;29:377-83.
9. Kaplan DL, Fossey S, Viney C, Muller W. Self-organization (as-
sembly) in biosynthesis of silk fibers—a hierarchical problem. In:
Aksay IA, Baer E, Sarikaya M, Tirrell DA, editors. Hierarchically
structured materials. Mater Res Symp Proc 1992;255:19-29.
10. Bai J, Ma T, Chu W, Wang R, Silva L, Michal C, et al.
Regenerated spider silk as a new biomaterial for MEMS. Biomed
Microdevices 2006;8:317-23.
11. Craig CL, Riekel C. Comparative architecture of silks, fibrous
proteins, and their encoding genes in insects and spiders. Comp
Biochem Physiol B Biochem Mol Biol 2003;135:721-2.
12. Lorena M, Robert F. Silk implants for the healing of critical size
bone defect. Bone 2005;37:688-98.
OOOOE
May 2010
13. Hirano Y, Mooney DJ. Peptide and protein presenting materials
for tissue engineering. Adv Mater 2004;16:17-25.
14. Dal Pra I, Freddi G, Minic J, Chiarini A, Armato U. De novo
engineering of reticular connective tissue in vivo by silk fibroin
nonwoven materials. Biomaterials 2005;26:1987-99.
15. Horan RL, Antle K, Collette AL, Wang Y, Huang J, Moreau
JE, et al. In vitro degradation of silk fibroin. Biomaterials
2005;26:3385-93.
16. Gosline JM, Demont ME, Denny MW. The structure and prop-
erties of spider silk. Endeavour 1986;10:37-43.
17. Mueller AA, Rahn BA, Gogolewski S, Leiggener CS. Early dural
reaction to polylactide in cranial defects in rabbits. Pediatr Neu-
rosurg 2005;41:285–21.
18. Nair MK, Nair UP, Seyedain A, Gassner R, Piesco N, Mooney
M, et al. Correlation of tuned aperture computed tomography
with conventional computed tomography for evaluation of osse-
ous healing in calvarial defects. Oral Surg Oral Med Oral Pathol
Oral Radiol Endod 2007;103:267-73.
19. Haddad AJ, Peel SA, Clokie CM, Sándor GK. Closure of rabbit
calvarial critical-sized defects using protective composite allogeneic
and alloplastic bone substitutes. J Craniofac Surg 2006;17:926-34.
20. Moghadam HG, Sandor GK, Holmes HH, Clokie CM. Histomor-
phometric evaluation of bone regeneration using allogeneic and
alloplastic bone substitutes. J Oral Maxillofac Surg 2004;62:202-13.
21. Stitzel J, Liu J, Lee SJ, Komura M, Berrya J, Sokerc S, et al.
Controlled fabrication of a biological vascular substitute. Bio-
materials 2006:27;1088-94.
22. Murugan R, Ramakrishna S. Development of nanocomposites
for bone grafting. Compos Sci Technol 2005;65:2385-406.
23. Rossitch EJr, Bullard DE, Oakes WJ. Delayed foreign-body
reaction to silk sutures in pediatric neurosurgical patients. Childs
Nerv Syst 1987;3:375-8.
24. Soong HK, Kenyon KR. Adverse reactions to virgin silk sutures
in cataract surgery. Ophthalmology 1984;91:479-83.
25. Zhao J, Zhang Z, Wang S, Sun X, Zhang X, Chen J, et al.
Apatite-coated silk fibroin scaffolds to healing mandibular bor-
der defects in canines. Bone 2009;45:517-27.
26. Li C. Electrospun silk-BMP-2 scaffolds for bone tissue engineer-
ing. Biomaterials 2006;27:3115-24.
27. Wang X, Kim HJ, Xu P, Matsumoto A, Kaplan DL. Biomaterial
coating by stepwise deposition of silk fibroin. Langmuir 2005;21:
11335-41.
28. Cai K, Yao K, Lin S, Yang Z, Li X, Xie H, et al. Poly(D, L-lactic
acid) surfaces modified by silk fibroin: effects on the culture of
osteoblast in vitro. Biomaterials 2002;23:1153-60.
29. Buchman SR, Sherick DG, Goulet RW, Goldstein SA. Use of
microcomputed tomography scanning as a new technique for the
evaluation of membranous bone. J Craniofac Surg 1998;9:48-54.
30. Cao Y, Wang B. Biodegradation of silk biomaterials. Int J Mol
Sci 2009;10:1514-24.
31. Postlethwait RW. Tissue reaction to surgical sutures. In: Dum-
phy JE, van Winkle W, editors. Repair and regeneration. New
York: McGraw-Hill; 1969. p. 263-85.
Reprint requests:
Dr. S. G. Kim
Department of Oral and Maxillofacial Surgery
College of Dentistry
Gangneung-Wonju National University
Jibyun-Dong, Gangneung
Gangwondo, 210-702
Korea
epker@chollian.net