Tài liệu hạn chế xem trước, để xem đầy đủ mời bạn chọn Tải xuống
1
/ 191 trang
THÔNG TIN TÀI LIỆU
Thông tin cơ bản
Định dạng
Số trang
191
Dung lượng
1,74 MB
Nội dung
Enhancing Resin/Dentin bond durability: The Effect
of Chitosan/Riboflavin Modification
By
Umer Daood
Thesis Submitted
In Partial Fulfillment of the Requirements
For the Degree of
MASTER OF SCIENCE (RSH-FoD)
At
Discipline of Oral Sciences, Faculty of Dentistry
National University of Singapore
2013
Supervisors: Assistant Professor Amr Fawzy (Main supervisor)
Associate Professor Cao Tong
!
"!
(Co- supervisor)
Declaration
I hereby declare that the thesis is my original work and it has
been written by me in its entirety. I have duly
acknowledged all the sources of information which have
been used in the thesis.
This thesis has also not been submitted for any degree in any
University previously.
_________________
Dr Umer Daood
20th November 2013
!
#!
Acknowledgement
My deepest appreciation to Almighty Allah for letting me through this
academic journey. I would like to thank my supervisor and the scientific committee
members on providing me full support for my work here at Faculty of Dentistry,
National University of Singapore. My special thanks to Dr Amr Fawzy for his
thoughtful and patient guidance throughout the course. I would also acknowledge my
co-supervisor Associate Professor Cao Tong who had also helped me get through
with this academic task. I am indebted for all the efforts that they have put in and
helping me achieve this milestone. I would also want to thank Dr. Sudhiranjan
Tripathy and Mr. Surani Dolmanan at Institute of Materials Research Engineering
(IMRE, Singapore) for their technical assistance in micro-Raman analysis of the
specimens.
My special acknowledgements to my wife, son and family whose support had
helped me throughout the entire course. Finally I would like to thank the members of
the lab and the friends in NUS who took me from strength to strength.
!
$!
Table of Contents
Acknowledgement -------------------------------------------------------------------
2
Table of Contents --------------------------------------------------------------------
3
!"#$%&'%(")*+,#!-----------------------------------------------------------------------
6
List of Tables ------------------------------------------------------------------------
42
List of Abbreviations ---------------------------------------------------------------
48
List of Deliverables ------------------------------------------------------------------
50
Abstract -------------------------------------------------------------------------------
51
1.
Introduction to Dentin Structure -------------------------------------------
54
1.1. Intertubular dentin --------------------------------------------------------------
55
1.2. Peritubular dentin ---------------------------------------------------------------
57
1.3. Tertiary dentin -------------------------------------------------------------------
58
2.
Riboflavin – Chemistry and Biological function -------------------------
59
3.
Collagen Structure -------------------------------------------------------------
63
3.1. Distribution and Biosynthesis --------------------------------------------------
64
3.2. Functions --------------------------------------------------------------------------
66
3.3. Degradation -----------------------------------------------------------------------
68
3.4. Bonding Hydrolysis --------------------------------------------------------------
69
4. Chitosan Structure --------------------------------------------------------------
70
4.1. pH ----------------------------------------------------------------------------------
71
4.2. Applications ----------------------------------------------------------------------
72
4.3. The Collagen Chitosan Relationship ------------------------------------------
73
5. Singlet Oxygen Radical Theory ----------------------------------------------
75
5.1.Riboflavin singlet oxygen --------------------------------------------------------
77
5.2. Generation of singlet oxygen by light in the presence of
!
%!
Endogenous and exogenous sensitizers ----------------------------------------------
78
5.3. Singlet oxygen and protein breakdown -----------------------------------------
78
6. Collagen Crosslinking -----------------------------------------------------------
80
7. Raman Spectroscopy ------------------------------------------------------------
84
7.1. Raman Mapping and Imaging Instrumentation ------------------------------
84
7.2. Sampling for Raman Spectroscopy; Applications ---------------------------
85
7.3. Protein Spectra -------------------------------------------------------------------
86
7.4. Raman of the Hybrid Layer -----------------------------------------------------
87
7.5. Raman spectroscopy is an effective technique for --------------------------the analysis of monomers and polymers
89
8. Resin bonding to dentin ---------------------------------------------------------
91
8.1. Adhesive systems ----------------------------------------------------------------
94
8.2.Two-Step-etch-and-rinse --------------------------------------------------------
95
8.3. Self-etch adhesive system – a drive for simplification----------------------
97
8.4. Hybrid layer formation and degradation ------------------------------------
98
8.5. Crosslinking and reinforcement of dentin collagen ------------------------
103
9. Hypotheses and Aim!------------------------------------------------------------
107
10. Materials and Methods -------------------------------------------------------
109
10.1. Phase I -------------------------------------------------------------------------
109
10.2. Phase II ------------------------------------------------------------------------
116
10.3. Phase III -----------------------------------------------------------------------
122
11. Results ---------------------------------------------------------------------------
127
11.1. Phase I -------------------------------------------------------------------------
127
11.2. Phase II ------------------------------------------------------------------------
130
11.3. Phase III -----------------------------------------------------------------------
133
!
&!
12. Discussion!-----------------------------------------------------------------------
137
12.1. Introduction -------------------------------------------------------------------
137
12.2. Phase I Discussion -----------------------------------------------------------
142
12.3. Phase II Discussion ----------------------------------------------------------
148
12.4. Phase III Discussion ---------------------------------------------------------
153
13. Summary, conclusions and Future Work ------------------------------
158
14. Bibliography ------------------------------------------------------------------
162
!
'!
List of Figures
Fig 1. SEM of 35% phosphoric acid etched dentin showing open dentinal tubules
lined with peritubular dentin as indicated by arrow (adapted from SEM evaluation of
the interaction pattern between dentin and resin after cavity preparation using
ER:YAG laser; Journal of Dentistry (2003) 31, 127–135).
!
(!
Fig 2. Schematic representation of the peritubular and mineralized intertubular dentin.
!
)!
Fig 3. Tertiary dentin also known as Reactive Dentin is seen clearly in this tooth
model produced as a reaction to the caries.
(http://www.dentalcaries.com/page.asp?pid=605).
!
*!
Fig 4. Schematic presentation of the chemical structure of riboflavin indicating the
CH2OH positioning by the transfer of electrons in alloxan, and oxylene.
!
"+!
Fig 5. Type I collagen shown as a molecular structure of fibrillar collagen with
various subdomains with cleavage sites for N- and C-procollagenases (adapted and
redrawn from ‘Collagens—structure, function, and biosynthesis’; Advanced Drug
Delivery Reviews 55 (2003) 1531– 1546)
!
""!
Fig 6. The lysyl mediated mature crosslinks formed within the collagen Type I fiber;
lysyl pyridinoline and hyroxylysyl-pyridinoline.
!
"#!
Fig 7. The collagen triple helix. (a) The crystal structure of collagen molecule; (b)
view down the axis of triple helix with three strands with space filling, ball stick and
ribbon presentation; (c) ball and stick profile of collagen triple helix; (d) stagger for
three strands (Proteins: Three Dimensional Structure; Section 6-1. Secondary
Structure).
!
"$!
Fig 8. The degree of N-acetylation in the physiochemical nature of chitin and chitosan
(adapted from Biomedical Activity of Chitin/Chitosan Based Materials—Influence of
Physicochemical Properties Apart from Molecular Weight and Degree of NAcetylation; Polymers 2011, 3(4), 1875-1901; doi:10.3390/polym3041875 Review).
!
"%!
Fig 9. Patterns of cross-linking collagens. Collagen types I (2), III (4), and IV (6)
show a banding pattern distinct from the other two shown. The riboflavin sensitization
with UVA causes the collagen Type I to almost disappear (3) [Effects of UltravioletA and riboflavin on the Interaction of Collagen and Proteoglycans during Corneal
Cross-linking; Published, JBC Papers in Press, February 18, 2011, DOI
10.1074/jbc.M110.169813; Yuntao Zhang1, Abigail H. Conrad, and Gary W. Conrad.
!
"&!
Fig 10. The Type I mechanism leading to electron transfer as a result of hydrogen
atom abstraction, thus yielding free radicals, which in turn can react with the available
oxygen species to form the superoxide ion. The Type II mechanism results in
collision of the excited sensitizer and the triplet excited oxygen that also results in an
energy transfer. [Reproduced from (1995) Royal Chemical Society].
!
"'!
Fig 11. The allysine crosslinking pathway with lysine residues for intermolecular
crosslinking. The skin collagen involves histidine forming mature crosslinks (Adapted
from Collagen Cross-Links; Top Curr Chem (2005) 247: 207–229).
!
"(!
Fig 12. Hydroxylation of crosslinking lysine residues showing bone tissue specific
crosslinking (Adapted from Collagen Cross-Links; Top Curr Chem (2005) 247: 207–
229).
!
")!
Fig 13. Raman and Rayleigh scattering compared in the Jablonski diagram in which a
molecule getting excited to a virtual state in lower energy with photon losing its
energy in Stokes Raman shift and gains a photon in Anti-Stokes Raman shift.
!
"*!
Fig 14. Raman spectra obtained from procine cartilage explants with minimal 960 cm1
peak [Adapted from Early detection of biomolecular changes in disrupted porcine
cartilage using polarized Raman spectroscopy; J. Biomed. Opt. 2011;16(1):017003017003-10. doi:10.1117/1.3528006].
!
#+!
Fig 15. Schematics (redrawn) showing acid etching of mineralized dentin removing
the smear layer leading to demineralization and exposing the collagen fibrils.
(Redrawn from Pashely DH, Ciucchi B, and Sano H. Dentin as a bonding substrate.
Dtsch Zahn Z 1994;49:760-63)
!
#"!
Fig 16. Scanning electron micrograph of the resin-dentin interface bonded with 3M
Single Bond ESPE (1500x). (a) dental composite; (b) adhesive bond; (c) hybrid layer;
(d) resin tags.
!
##!
Fig 17. Scanning electron microscopic image of the resin-dentin interface bonded
with 3M Single Bond ESPE. A uniform hybrid layer (HL) formation can be seen with
visible resin tag (RT) formation.
!
#$!
Fig 18. Collagenolysis in the presence of collagenase and unwinding of the triple
helix in the alpha chain (Nagase H, Fushimi K. Elucidating the function of noncatalytic domains of collagenases and aggrecanases. Connective Tissue Research.
2008;16:9:74).
!
#%!
Fig 19. Obtained Raman spectrum of the (a) control demineralized specimen; (b)
Adper TM Single Bond; (c) and chitosan in the region of 700-1700 cm-1 (Daood et al;
Effect of chitosan/riboflavin modification on resin/dentin interface: Spectroscopic and
microscopic investigations).
!
#&!
Fig 20. Schematic representation of the specimen preparation for the micro-tensile
bond strength (!TBS) test and SEM analysis. (A) Removal of occlusal surface to
expose the flat dentin surface; (B) preparation of the dentin surface to receive
adhesive with or without RF; (C) cutting of resin-dentin beams from the center of
bonded specimen; (D) attachment of the single resin-dentin beam for immediate
!TBS testing; (E) storage of resin-dentin beams in artificial saliva for 9-months
storage for !TBS testing and SEM analysis.
!
#'!
Fig 21. Bonded specimen glued to custom-made metallic jig mounted to Universal
testing machine with cyanoacrylate adhesive.
!
#(!
Fig 22. Representative Raman spectra of (A) Adper TM Single bond adhesive, (B)
demineralized dentin specimen, and (C) resin impregnated dentin recorded in the
region between 800-1800 cm-1. The P-O bond at 960 cm-1of the mineral component
is well represented for demineralized and resin impregnated dentin. The peaks at 1667
cm-1 and 1246 cm-1 are associated with organic components for dentin collagen.
!
#)!
Fig 23. Representative SEM micrographs of the etched dentin resulting from different
bio-modification procedures. Images showing an illustrative area of the dentin surface
of (A) control; bio-modified with (B) 0.1%RF; (C) Ch/RF 1:4 and (D) Ch/RF 1:1
specimens. The 0.1%RF and Ch/RF 1:4 specimens show open dentinal tubules with
intact collagen fibers whereas the Ch/RF 1:1 specimens exhibit a discontinuous
structure. The dentin of all specimens were conditioned for 15 s with 37% phosphoric
acid (Daood et al; Effect of chitosan/riboflavin modification on resin/dentin interface:
Spectroscopic and microscopic investigations).
!
#*!
Fig 24. SEM images of the control, 0.1% RF and Ch/RF (1:4 and 1:1) crosslinked
resin/dentin interfaces surfaces treated with AdperTM Single bond 2; 3M ESPE. The
hybrid layer (HL) and many resin tags (RT) were found at the adhesive interface
between resin cement and dentin; (A) control, (B) A uniform hybrid layer with long
resin tags can be observed in the specimen interface treated with 0.1%RF crosslinking
prior to dentin bonding agent application. The resin cement penetrated deeply and
many long resin tags were observed at the demineralized interface. (C) A funnelshaped configuration of the resin tags also seen at the base of Ch/RF 1:4-treated
specimens. The resin tags exhibited a slightly rough texture. (D) The resin tags in
Ch/RF 1:1 specimens showed a regular cylindrical shape exhibiting a rough texture
(arrow) on top of the resin tags and a relatively thicker and more textured hybrid
layer; HL, hybrid layer; RT, resin tags (Daood et al; Effect of chitosan/riboflavin
modification on resin/dentin interface: Spectroscopic and microscopic
investigations).
!
$+!
Fig 25. Raman spectra of control, 0.1%RF and Ch/RF (1:4 and 1:1) crosslinked dentin
specimens in the spectral range of 900–1700 cm-1. The peak assignments are
represented in Table II. The C-H alkyl groups also appeared in the collagen spectrum
in dentinal substrate with other peak areas of CAC bond, Amide I and Amide III.
Schematic representation of CH3 inplane bending also shown. (a) control (b) 0.1%RF
(c) Ch/RF 1:4 (d) Ch/RF 1:1. [Color figure can be viewed in the online issue, which is
available at wileyonlinelibrary.com ; Daood et al; Effect of chitosan/riboflavin
modification on resin/dentin interface: Spectroscopic and microscopic
investigations].
!
$"!
Fig 26. Raman line map (A) and images of the spectrum at the 4 !m (B) and 8 lm (C)
crosslinked resin/dentin interfaces. Intensities at 960 cm and 1450 cm-1 for all other
specimens are identified in the line map. The Raman images indicate the positions of
spectra in the region of interest. The spectrum is characterized to (a) Ch/RF 1:1 (b)
Ch/RF 1:4 (c) 0.1%RF (d) control. [Color figure can be viewed in the online issue,
which is available at wileyonlinelibrary.com. Daood et al; Effect of
chitosan/riboflavin modification on resin/dentin interface: Spectroscopic and
microscopic investigations].
!
$#!
Fig 27. SEM images of the resin-dentin interface after 24 h storage in artificial saliva.
Well-defined uniform hybrid layer (white arrows) and with well-formed branched
resin tags could be observed with control (A), 1%RF-modified adhesive (B), and
3%RF-modified adhesive (C) specimens. Relatively thick and textured hybrid layer
with long well formed resin tags could be seen with the 5%RF-modified adhesive
specimens (D). The 10%RF-modified adhesive specimens showed a very thin hybrid
layer with lack of well-formed resin tags (E). C: resin composite; HL: hybrid layer;
RT: resin tags.
!
$$!
Fig 28. SEM images showing the resin-dentin interface of the RF-modified adhesive
system after 9-months aging period in artificial saliva. Relatively intact hybrid layer
could be seen after 9-months storage for the control (A), 1% (B) and 3%RF-modified
specimens (C) compared to the 5%RF-modified specimens (D). Hardly any hybrid
layer could be observed in the 10%RF-modified specimens and only few short resin
tags could be seen (E).
!
$%!
Figure 29: Distribution (%) of failure mode of control and RF-modified adhesive
specimens after the micro-tensile strength testing of the immediate (A) and the 9months stored specimens (B) in artificial saliva.
!
$&!
Fig 30. Representative fracture surfaces of specimens bonded with control and RFmodified adhesives. Control group (A), without RF, showing a typical mixed fracture
pattern at the outer rim, occurring mostly at the bottom of the hybrid layer (higher
magnification shown in solid box); mixed failure in 1%RF-modified adhesive
specimens with open dentinal tubules, and adhesive remnants (arrow) (B); 3%RFmodified adhesive specimens, with mixed failure pattern and with resin tags (C and
D;); resin tags within the dentinal tubules of 3%RF-modified adhesive specimens (E;
open arrows); 5%RF-modified adhesive specimens at lower magnification presenting
dentin side of fracture with mixed failure pattern (solid box) and cohesive failure
within the adhesive (dotted box) (F); adhesive failure within 10%RF-modified
adhesive specimens (G); small cracks within 5%RF-modified adhesive specimens
after 9-months of artificial saliva (H; arrows); Cohesive failure in 10%RF-modified
adhesives (A=adhesive) after 9-months of storage (I). Horizontal fracture seen at the
interfacial region in 10%RF-modified adhesive specimens after 9-months of storage
(J).
!
$'!
Fig 31. The percentage of degree of conversion of control and different RF-modified
adhesive specimens at different time intervals from the start of photoactivation (0 s)
till 30 minutes.
!
$(!
Fig 32. Representative Raman spectrum for the five adhesives tested illustrating the
principal functional groups. The groups can be identified at CH2CH3 1450 cm-1, C=O
1610 cm-1, C=O 1640 cm-1 and C=O 1720 cm-1. The arrows indicate shifts associated
with C=O 1720 cm-1;(A) control=1717 cm-1, (B) 1%RF-modified adhesive=1711 cm1
, (C) 3%RF-modified adhesives=1710 cm-1, (D) 5%RF-modified adhesives =1713
cm-1 and (E) 10%RF-modified adhesives = 1727 cm-1.
!
$)!
Fig 33. Representative line map (scans) across resin-dentin interface of different
control and riboflavin-modified adhesive specimens. The spectral contribution is
recorded at 960 cm-1 (hydroxyapatite) and 1450 cm-1 (C-H Alkyl) intensities
representing the penetration of different adhesives.
!
$*!
Fig 34. Comparison of Raman spectral data acquired in the region of 1030 cm-1 and
1670 cm-1 for the (A) Control, (B) 1%, (C) 3%, (D) 5%, and (E) 10%RF-modified
adhesive specimens at 5!m levels. The principal bands identified are Amide bands (I
and III) along with C-H alkyl groups in the resin-dentin specimens. The single arrow
indicates the pyridinium ring group where accentuated intensity is observed in 3%RFmodified adhesive specimen.
!
%+!
Fig 35. Calculated ratios of 1610 cm-1 to 1640 cm-1 suggesting the ratio of aromatic to
aliphatic groups.
!
%"!
Fig 36. (A) Confocal fluorescence images of the resin-dentin interfaces of (i) control,
(ii) 1%, (iii) 3%, (iv) 5%, and (v) 10%RF-modified two-step etch-and-rinse adhesive
specimens. The image shows variation in micro permeability within the interface. (B)
3D image of resin tags formed by 3%RF-modified adhesive specimens.
!
%#!
List of Tables
%
,-./0!"1!Data representing the proportionate mixture of Chitosan and Riboflavin for
crosslinking treatment.%
-.%/&.%!
01"$/%2/3%4"5&'627"/%8&/8,/$+2$"&/%"/%$,+9#%&'%!
!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!2345/!!!!!!!!!!!!!!!!!!!!!!!!!!!6745/!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!6-89:!:;!23467!590!
"1!
#1!
&+!
#+!
&+!
)+!
"?"!!
"?%!
!
@//!A-5B/0A!C0>0!D>:AA/9EF0G!C983!+1"H!67!-EG!:>!23467!I"?%J!"?"K!;:>5=/-89:EA!-A!G0AD>9.0G!9E!830!L-80>9-/A!-EG!
L083:GA!
!
%$!
Table 2. Typical band assignments of Raman spectrum of dentin collagen disc
specimens in both innate state and after cross-linking treatment of dentin disc
specimens.
!
!
!
0&662),/!
:.;bond strength
RF
Riboflavin - riboflavin
Ch
Chitosan
CaCl2
Calcium chloride
KCl
Potassium chloride
MgCl2 Magnesium chloride
H2O
Water
sec
Seconds
SiC
Silicon carbide
kV
kilovolt amperes
CPD
Critical point drying
MPa
Mega Pascals
$L
micro liter
DOC
Degree of conversion
CLSM Confocal light scanning microscopy
wt % Weight percent
ɛ%%%%%%%%%%%%%AC#"6&/%
+C9%%%%%%%4&$2$"&/%C,+%9"/*$,
!
&+!
List of Deliverables
1. Daood U, Iqbal K, Nitisusanta LI, Fawzy AS. Effect of chitosan/riboflavin
modification on resin/dentin interface: spectroscopic and microscopic investigations. J
Biomed Mater Res A. 2013 Jul;101(7):1846-56. doi:10.1002/jbm.a.34482. Epub 2012
Nov 27.
2. Daood U, Heng CS, Lian JNC, Fawzy AS. Riboflavin-modified experimental twostep etch-and-rinse dentin adhesive: an in vitro Study. Int J Oral Sc Manuscript #
IJOS201307302; under review)
3. Daood U, Fawzy AS. Investigation of the resin-dentin interface using riboflavinmodified adhesive: Raman and Confocal microscopy analysis. Int Dent J Manuscript
# IDJ-Jul-13-OA-0256; under review)
4. Poster presentation in FDI Annual World Dental Congress 2012, Hong Kong.
Theme: Dental Materials and Restorative Dentistry – Materials. P112 - Microscopic
and Spectroscopic Characterization of the Effect Chitosan/Riboflavin Modification on
Dentin/Resin Interface. Umer Daood, Lorraine Ivana Nitisusanta, Kulsum Iqbal,
Jennifer Neo Chiew Lian, Amr Fawzy.
(http://www.fdicongress.org/c/document
library/get file?uuid=aeb4e370-36e7-453e-ba9a-45c469004a16&groupId=27627)
!
&"!
Abstract
Objectives: The aim of this study is to investigate the morphological and chemical
changes of demineralized dentin collagen-matrix and resin/dentin interface associated
with chitosan/riboflavin modification. Two-step experimental etch-and-rinse model
dentin adhesive was also modified with different concentrations of riboflavin and
study its effect on the bond strength and on the degree of conversion of the adhesive.
In addition, investigation of the modification of commercially available two-step etchand-rinse adhesive with different concentrations of riboflavin was performed to study
the effect of riboflavin modification on resin impregnation. Materials and Methods:
In phase I of the study, dentin disc specimens were prepared from sound molars, acidetched with 35% phosphoric acid and modified with either 0.1% riboflavin or
chitosan/riboflavin (Ch/RF ratios 1:4 or 1:1) and photo-activated by UVA. Dentin
surfaces of sound molars were exposed, acid-etched, and modified as described
before. Etch-and-rinse dentin adhesive was applied, light-cured, and layered with
resin-restorative composite. The resin infiltration and resin/dentin interface were
characterized by micro-Raman spectroscopy and SEM. Moreover, in phase II, an
experimental adhesive-system was modified with different concentrations of
riboflavin (0, 1, 3, 5 and 10 wt%). Similarly, dentin surfaces were etched with 37%
phosphoric acid, bonded with respective adhesives, restored with restorative
composite-resin, and sectioned into beams to be stored for 24 hour or 9-months in
artificial saliva. Micro-tensile bond testing was performed along with scanning
electron microscopy to analyze the failure distribution of debonded beams and to
investigate resin-dentin interface morphology. The degree of conversion was
evaluated by performing Fourier transform infrared spectroscopy at different timepoints from the start of irradiation. Data was analyzed with one-way and two-way
!
!
ANOVA followed by Tukey’s for pair-wise comparison. In phase III of the study,
commercially available AdperTM Single Bond adhesive-system was modified with
different concentrations of riboflavin (1, 3, 5 and 10 wt%) or left unmodified as a
control. Dentin surfaces were again etched with 37% phosphoric acid, bonded with
two coats of respective adhesives and restored with restorative composite-resin. The
resin infiltration and resin-dentin interface were characterized by micro-Raman
spectroscopy. Confocal microscopy was performed by adding Rhodamine B dye
(0.01%) to the bonding resin. Data was analyzed with two-way ANOVA followed by
Tukey’s for pair-wise comparison. Results: In Phase I of the study, an open-intact
collagen network-structure, formation of uniform hybrid-layer and higher resin
infiltration were found with 0.1%RF and Ch/RF 1:4 modifications. Raman analysis
revealed chemical changes and shifts in Amide bands with the modification of dentin
collagen-matrix. Results in Phase II of the study, modification with 1% and 3%
riboflavin increased the micro-tensile bond strength compared to the control at 24 h
and 9-months storage with no significant differences in degree of conversion
(pD30A9!,\!]0983!P!W\!^->8:E!@1!!_!@G30A9:E1!"**#J$*?")&1!
!
"$( !
ASTM (American Society for Testing and Materials). Standard terminology of
adhesives. D907–70. Philadelphia, PA, 1970.
138
Buonocore MG. A simple method of increasing the adhesion of acrylic filling
materials to enamel surfaces. J Dent Res. 1955;34:849-853.
!
!
"('!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
139
Sheth P, Jensen M, Sheth J. Comparative evaluation of three resin inlay
techniques: Microleakage studies. Quint Int. 1989;20:831-836.
140
Tay FR, Gwinnett JA, Wei SH. Micromorphological spectrum from overdrying to
overwetting acid-conditioned dentin in water-free acetone-based, single-bottle
primer/adhesives. Dent Mater. 1996;12:236-44.
141
Van Landuyt KL, Snauwaert J, De Munck J, Peumans M, Yoshida Y, Poitevin A.
Systematic review of the chemical composition of contemporary dental adhesives.
Biomaterials. 2007;28(26):3757-85.
142
Tay FR, Pashley DH, Suh BI, Hiraishi N, Yiu CK. Water treeing in simplified
dentin adhesives-deja vu? Oper Dent. 2005;5:561-79.
143
Tay FR, Pashley DH, Suh BI, Carvalho RM, Itthagarun A. Single step adhesives
are permeable membranes. J Dent. 2002;30:371-82.
144
Carvalho RM, Carrilho MRO, Pereira LCG, Marquezini L Jr, Silva SMA,
Kussmaul APM. Adhesive systems: foundations for clinical application. Revista
Biodonto. 2004;2:6-86.
145
Van Meerbeek B, Vargas M, Inoue S, Yoshida Y, Peumans M, Lambrechts P,
Vanherle G. Adhesives and cements to promote preservation dentistry. Oper
Dent;6:119–144, 2001.
146
Bastioli C, Romano G, Migliaresi C. Water sorption and mechanical properties of
dental composites. Biomater;11: 219–223, 1990.
147
Yoshida Y, Nagakane K, Fukuda R, Nakayama Y, Okazaki M, Shintani H.
Comparative study on adhesive performance of functional monomers. J Dent Res.
2004;83:454–458.
148
Breschia L, Mazzonib A, Ruggerib A, Cadenaroa M, Lenardaa R D, Dorigo E D S.
Dental adhesion review: Aging and stability of the bonded interface. Dent Mat.
!
"((!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
2008:2 4;90–101.
149
Masuhara E, Kojima K. Hirasawa T, Tanuni N, Kimura T. Studies on dental self-
curing resins on the adhesive force to ivory and tooth surface. Rep Res Inst Dent
Mater. 1963:2:457-465.
150
Tay F R, Pashley DH. Water treeing–a potential mechanism for degradation of
dentin adhesives. Am J Dent. 2003;1:6-12.
151
Tjäderhane L, Larjava H, Sorsa T. The activation and function of host matrix
metalloproteinases in dentin matrix breakdown in carious lesions. J Dent Res. 1998.
778:1;622-1629.
152
Pashley D H, Tay F R, Yiu C. Collagen degradation by host-derived enzymes
during aging. J Dent Res. 2004. 83;3:216-221.
153
Perumal S, Antipova O, Orgel J P. Collagen fibril architecture, domain
organization, and the triple helical confirmation govern its proteolysis. Proceedings of
the National Academy of Sciences of the United States of America. 2008;105:2824-9.
154
Mazzoni A, Breschi L, Carrilho M, Nascimento FD, Orsini G, Ruggeri A. A
review on nature, role and functions of dentin non-collagenous proteins. Part
II:enzymes, serum proteins and growth factors. Endodontic topics. 2012;21:19-40.
155
Mazzoni A, Pashley D H, Nishitani Y, Breschi L, Mannello F, Tjaderhane L.
Reactivation of inactivated endogenous proteolytic activities in phosphoric acid
etched dentin by etch-and-rinse adhesives. Biomaterials. 2006;27:4470-4476.
156
Saftig P, Hunziker E, Wehmeyer O, Jones S, Boyde A, Rommerskirch W.
Imparied Osteoclastic bone resorption leads to osteopetrosis in Cathepsin K deficient
mice. Proceedings of the National Academy of Sciences of the United States of
America. 1998;95:13453-13458.
!
"()!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
157
Carrilho R O, Carvalho R M, Tay F R, Yiu C, Pashley D H. Durability of resin–
dentin bonds related to water and oil storage. Am J Dent. 2005;18:315–319.
158
Visse R, Nagase H. Matrix metalloproteinases and tissue inhibitors of
metalloproteinases, structure, function and biochemistry. Circ Res. 2003;92:827–39.
159
Tay F R, Pashley D H. Have dentin adhesives become too hydrophilic? J Can Dent
Assoc. 2003;69:726–31.
160
Strijp A J, Jansen D C, DeGroot J, ten Cate J M, Everts V. Host derived
proteinases and degradation of dentin collagen in situ. Caries Res. 2003;37;58-65.
161
Scott J E, Thomlinson A M. The structure of interfibrillar proteoglycan bridges
shape modules in extracellular matrix of fibrous connective tissues and their stability
in various chemical environments. J Anat. 1998;192:391–405.
162
Breschi L, Mazzoni A, Ruggeri A, Cadenaro M, Di Lenarda R, Dorigo E. Dental
adhesion review: Aging and stability of the bonded interface. Dent Mater.
2008;24:90–101.
163
Manuja N, Nagpal R, Pandit IK. Dental Adhesion: Mechanism, Techniques and
Durability. J Clin Pediat Dent. 2012;36:223-234.
164
Hjeljord LG, Rolla G, Bonesvoll P. Chlorhexidine-protein interactions. J Periodon
Res. 1973;12:11–16.
165
Cheung DT, Tong D, Perelman N, Ertl D, Nimni ME. Mechanism of crosslinking
of proteins by glutaraldehyde. IV: In vitro and in vivo stability of a crosslinked
collagen matrix. Connect Tissue Res. 1990;25:27–34.
166
A. Tezvergil-Mutluay, Mutluay M, Agee KA, Seseogullari-Dirihan R, Hoshika T,
Cadenaro M, Breschi L, Vallittu P, Tay FR, Pashley DH. Carbodiimide Cross-linking
Inactivates Soluble and Matrix-bound MMPs, in vitro. J Dent Res, 2012; 91, 192.
!
"(*!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
167
Cova A, Breschi L, Nato F, Ruggeri A, Carrilho M, Tjäderhane L, Prati C,
Lenarda D, Tay FR, Pashley DH. Effect of UVA-activated Riboflavin on Dentin
Bonding, J Dent Res, 2011; 90, 1439.
"') !HSU
YS, Jordan TH, Dederich DN. Effects of low energy CO2 laser irradiation
and the organic matrix on inhibition of enamel demineralization. J Dent Res.
2000;79:1725-1730.!
"'* !
Priestley CM, Williamson EM, Wafford KA. Thymol, a constituent of thyme
essential oil, is a positive allosteric modulator of human GABA (A) receptors and a
homo-oligomeric
GABA
receptor
from Drosophila
melanogaster. Br
J
Pharmacol. 2003;140:1363–1372.
170
Rinaudo M, Pavlov G, Desbrieres J. Influence of acetic acid concentration on the
solubilization of chitosan. Polymer. 1999;40: 7029–7032.
171
Seda R, Aydin T, Pulat M. 5-Fluorouracil encapsulated chitosan nanoparticles for
pH-stimulated drug delivery: Evaluation of controlled release kinetics. J Nanomater
2012;42:3124.
172
Stossel P, Leuba JL. Effect of chitosan, chitin and some aminosugars on growth of
various soil borne phytopathogenic fungi. Phytopathol Z. 1984;111:82–90.
173
Xu C, Yao X, Walker MP, Wang Y. Chemical/molecular structure of the dentin–
enamel junction is dependent on the intratooth location. Calcif Tissue Int
2009;84:221–228.
174
Miyazakia M, Onosea H, Mooreb BK. Analysis of the dentin-resin interface by use
of laser Raman spectroscopy. Den Mater. 2002;18:576–580.
175
Ye Q, Spencer P, Wang Y, Misra A. Relationship of solvent to the
photopolymerization process, properties, and structure in model dentin adhesives. J
Biomed Mater Res A 2007; 80:342-350.
!
")+!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
176
Park J, Ye Q, Topp EM. Effect of Photoinitiator System and Water Content on
Dynamic Mechanical Properties of a Light-cured bisGMA/HEMA Dental Resin. J
Biomed Mater Res A. 2010;93:1245-1251.
177
Ari H, Donmez N, Belli S. Effect of Artificial Saliva Contamination on Bond
Strength to Pulp Chamber Dentin. Eur J Dent. 2008;2:86–90.
178
Heymann HO, Bayne SC. Current concepts in dentin bonding: focusing on
dentinal adhesion factors. J Am Dent Assoc 1993;124(5):26-36.
"(* !
Fawzy A S, Amer M A, El-Askary F S. Sodium hypochlorite as dentin
pretreatment for etch-and-rinse single-bottle and two-step self-etching adhesives:
Atomic force micoscope and tensile bond strength evaluation. J Adhes Dent.
2008;10:135-144.!
")+ ! Spanberg
L. Biologic effects of dental materials. III. Toxicity and antimicrobial
effect of endodontic antisepticsin vitro. Oral Surg .1973;36:856 –71.!
")" !
14- Johal S, Baumgartner JC, Marshall JG. Comparison of the antimicrobial
efficacy of 1.3% NaOCl/BioPure MTAD to 5.25% NaOCl/15% EDTA for root canal
irrigation. J Endod. 2007;33:48-51.
")# !Duarte
R M, de Goes M F, Montes M A. Effect of time on tensile bond strength of
resin cement bonded to dentine and low-viscosity composite. J Dent. 2006;34:52-61.!
")$ !Loguercio
A D, Uceda-Gomez N, Carrilho M R. Influence of specimen size and
regional variation on long-term resin-dentin bond strength. Dent Mater 2005;21:224231.!
")% !
Nikaido T, Kunzelmann KH, Chen H. Evaluation of thermal cycling and
mechanical loading on bond strength of a self-etching primer system to dentin. Dent
Mater. 2002;18:269-275
!
")"!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
185
Epasinghe DJ, Yiu CK, Burrow MF. Effect of proanthocyanidin incorporation into
dental adhesive resin on resin–dentine bond strength. J Dent. 2012; 40(3):173-180.
186
Yoshida T, Yamaguchi K, Tsubota K. Effect of Metal conditioners on
polymerization behaviour of bonding agents. J Oral Sci. 2005; 47:171-175.
187
Heigl JJ, Bell MF, White JN. Application of Infrared Spectroscopy to the Analysis
of Liquid Hydrocarbons. Anal Chem 1947;19(5):293-298.
188
Jose P, Sakhamuri S, Sampath V, Sanjeev K, Sekar M. Degree of conversion of
two dentin bonding agents with and without a desensitizing agent using Fourier
transform infrared spectroscopy: An in vitro study. J Conserv Dent. 2011;14(3): 302305.
")* ! Schulein
TM. Infection control for extracted teeth in the teaching laboratory. J
Dent Educ. 1994;58:411-413.
"*+ !Miniotis
NJ, Bennett PS, Johnston AD. Molar efficiency study of chlorinated NPG
substitutes in dentin bonding. J Dent Res. 1993;72:1045-1049.
191
Xu C, Yao X, Walker MP, Wang Y. Chemical/molecular structure of the dentin-
enamel junction is dependent on the intratooth location. Calc Internat. 2009;84:221227.
192
D’souz PD, Duschner H, Staehle HJ Pioch T. Dentin bonding systems: a
comparative study of SEM and CLSM used to visualize the resin-dentin interface.
Acta Med Dent Helv. 1999;4:20-27.
193
Xu C, Wang Y. Collagen cross-linking increases its biodegradation resistance in
wet dentin bonding. J Adhes Dent. 2012;14:11-8.
194
Ozaki M, Suzuki M, Itoh K, Wakumoto S, Hisamitsu H. Laser Raman
spectroscopic study of the adhesive interface; analysis between 4-META/MMA-TBB
resin and bovine or human dentin. Dent Mater J. 1992;11:70-76.
!
")#!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
195
Urbanczyk GW, Symonowicz BL. The influence of processing terms of chitosan
membranes made of differently deacetylated chitin on the crystalline structure of
membranes. J of App Poly Sc. 1994;51:2191–2194.
196
Singla AK, Chawla M. Chitosan: some pharmaceutical and biological aspects—an
update. Pharm Pharmacol. 2001;53:1047–67.
197
Majeti NV, Kumar R. A review of chitin and chitosan applications. Reactive and
Functional Polymers. 2000;46:1–27.
198
Gingras M, Paradis I, Berthod F. Nerve regeneration in a collagen-chitosan tissue-
engineered skin transplanted on nude mice. Biomater. 2003;24:1653–1661.
199
Taravel MN, Domard A. Collagen and its interactions with chitosan. II. Influence
of the physicochemical characteristics of collagen. Biomater. 1995;16:865–871.
200
Wang XH, Li DP, Wang WJ, Feng QL, Cui FZ, Xu YX. Crosslinked
collagen/chitosan matrix for artificial livers. Biomater. 2003;24:3213-3220.
201
Wallace DG, Rosenblatt J. Collagen gel systems for sustained delivery and tissue
engineering. Adv Drug Deliv Rev 2003;55:1631–1649.
202
Han B, Jaurequi B, Tang BW. Proanthocyanidin: a natural crosslinking reagent for
stabilizing collagen matrices. J Biomed Mater Res Part A. 2003;65:118–124.
203
Choe E, Min DB. Chemistry and reactions of reactive oxygen species in foods.
Crit Rev Food Sci Nutr 2006;46:1–22.
204
Slifkin MA. Interaction of amino acids with riboflavin. Nat 1963; 197(4684): 275–
276.
205
Matheson IB, Etheridge RD, Kratowich NR et al. The quenching of singlet oxygen
by amino acids and proteins. Photo Photobl. 1975; 21(3):165–171.
206
Fawzy A, Nitisusanta L, Iqbal K, Daood U, Beng LT, Neo J. Characterization of
Riboflavin-modified Dentin Collagen Matrix. J Dent Res. 2012;91:1049-1054.
!
")$!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
207
Fawzy A, Nitisusanta L, Iqbal K, Daood U, Neo J. Riboflavin as a dentin
crosslinking agent: Ultraviolet A versus blue light. Dent Mater 2012;28:1284-1291.
208
Bellamy LJ. The Infrared Spectra of Complex Molecules, Vol.1, 3rd ed., London:
Chapman and Hall, pp. 37-148, 1975.
209
Spencer P, Wang Y, Walker MP, Wieliczka DM, Swafford JR. Interfacial
chemistry of the dentin/adhesive bond. J Dent Res. 2000;79:1458-1463.
210
Wieliczka DM, Kruger MB, Spencer P. Raman imaging of dental adhesive
diffusion. Appl Spect. 1997;51:1593-1596.
211
Meerbeek BV, Mohrbacher H, Celis JP, Roos JR, Braem M, Lambrechts L,
Vanherle G. Chemical characterization of the resin- dentin interface by micro-Raman
spectroscopy. J Dent Res 1993;72:584-585.
212
Hanlon EB, Monoharan R, Koo TW, Shafer KE, Motz JT. Prospects for in vivo
Raman spectroscopy. Phys Med Biol. 2000;45:1-59.
213
Meerbeek V, Dhem A, Goret-Nicaise M, Braem M, Lambrechts P, Vanherle G.
Comparative SEM and TEM examination of the ultrastructure of the resin-dentin
inter- diffusion zone. J Dent Res. 1993;72:495-501.
214
Eick JD, Robinson SJ, Byerle TJ, Chappel RP, Spencer P, Chappelo CC. Scanning
transmission electron microscopy/energy-dispersive spectroscopy analysis of the
dentin adhesive interface using a labeled 2-hydroxyethylmethacrylate analogue. J
Dent Res. 1995;74:1246-1252.
215
Griffiths BM, Naasan M, Sherriff M, Watson TF. Variable polymerization
shrinkage and the interfacial micro-permeability of a dentine bonding system. J Adhes
Dent. 1999;1:119-131.
#"' !Matousek
!
P, Clark IP, Draper ERC, Morris MD, Goodship AE, Everall N, Towrie
")%!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
M, Finney WF, Parker AW: Subsurface Probing in Diffusely Scattering Media Using
Spatially Offset Raman Spectroscopy. Applied Spectroscopy. 2005.59:393-400.
#"( ! Gyeong-Bok
J, Hui Jae L, Ji Hye K. Effect of crosslinking with riboflavin and
ultraviolet A on the chemical bonds and ultrastructure of human sclera. J Biomed
Opt;16:345-352.!
#") ! Strawn,
S.E., White, J.M., Marshall, S.J. Characterization of dentin after short-
term storage. J. Dent. Res. 1993. 72;8:383-389.
#"* !Goodis
H E, Marshall Jr G W, White J M. The effects of storage after extraction of
the teeth on human dentine permeability in vitro. Arch Oral Biol. 1991;36:561-566.
##+ ! Titley
K C, Chernecky R, Rossouw P E, Kulkarni G V. The effect of various
storage methods and media on shear-bond strengths of dental composite resin to
bovine dentine. Arch Oral Biol. 1998; 43: 305-11.
221
Crim GA, Mattingly SL. Evaluation of two methods for assessing marginal
leakage. J Prosthet Dent. 1981;45:160-3.
222
Crim GA, Swartz ML, Phillips RW. Comparison of four thermocycling
techniques. J Prosthet Dent. 1985;53:50-3.
223
Trowbridge H O. Model systems for determining biologic effects of microleakage.
Oper Dent. 1987;12:164-172
224
Wendt S L, McInnes P M, Dickinson G L. The effect of thermocycling in
microleakage analysis. Dent Mater. 1992;8:181-4.
225
Gale M S, Darvell B W. Thermal cycling procedures for laboratory testing of
dental restorations. J Dent. 1999;27:89-99.
226
Linde A. Dentin matrix proteins: Composition and possible functions in
calcification. Anat Rec. 1989;224:154–166.
227
!
Raspanti M, Ortolani F, Ruggeri A. Collagen fibril surface structures: Freeze")&!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
etching data and computer modeling. Int J Biol Macro. 1989;11:77–80.
228
Grover CN, Cameron RE, Best SM. Investigating the morphological, mechanical
and degradation properties of scaffolds comprising collagen, gelatin and elastin for
use in soft tissue engineering. J Mech Behav Biomed Mater. 2012;10:62–74.
229
Jung GB, Lee HJ, Kim JH, Lim JK, Park HK. Effect of cross-linking with
riboflavin and ultraviolet A on the chemical bonds and ultrastructure of human sclera.
J Biomed Opt 2011;16:125004.
230
Changyou LM, Zhengwei M, Zhou J, Shen J. Enhanced biological stability of
collagen porous scaffolds by using amino acids as novel cross-linking bridges.
Biomaterials. 2004;25:2997–3004.
231
Russo AKB, Castellan CS, Shinohara MS, Hassan L, Antunes A. Characterization
of biomodified dentin matrices for potential preventive and reparative therapies. Acta
Biomater. 2011;7:1735–1741.
232
Wollensak G, Spoerl E. Collagen crosslinking of human and porcine sclera. J
Cataract Refract Surg. 2004;30:689–695.
233
Tjaderhane L, Larjava H, Sorsa T, Uitto VJ, Larmas M, Salo T. The activation and
function of host matrix metalloproteinases in dentin matrix breakdown in caries
lesions. J Dent Res. 1998;77:1622–1629.
234
Silva CW, Whiting M, Lamoureux E, Lindsay RG, Sullivan LJ, Snibson GR. A
randomized controlled trial of corneal collagen cross-linking in progressive
keratoconus: Preliminary results. J Refract Surg. 2008;24:720–725.
235
Zhang J, Xia W, Liu P, Cheng Q, Tahirou T, Gu W, Li B. Chitosan modification
and pharmaceutical/biomedical applications. Mar Drugs. 2010;8:1962–1967.
236
Yan J, Yang L, Wang G, Xiao Y, Zhang B, Qi N. Biocompatibility evaluation of
chitosan-based injectable hydrogels for the culturing mice mesenchymal stem cells in
!
")'!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
vitro. J Biomater Appl. 2009; 24:625–637.
237
Ma L, Gao C Y, Mao Z W, Shen J C, Hu H X, Han C M. Thermal dehydration
treatment and glutaraldehyde cross-linking to increase the biostability of collagenchitosan porous scaffolds used as dermal equivalent. J Biomater Sci Polym.
2003;14:861–874.
238
Spencer P, Swafford JR. Unprotected protein at the dentin– adhesive interface.
Quint Int. 1999;30:501–507.
239
Pashley DH. Smear layer: Overview of structure and function. Proc Finn Dent Soc.
1992;88:215–224.
240
Fawzy AS. Variations in collagen fibrils network structure and surface dehydration
of acid demineralized intertubular dentin: Effect of dentin depth and air-exposure
time. Dent Mater. 2010;26:35–43.
241
Mehrdad R, Li F, Fagerholm P, Lagali N S, Watsky M A, Griffith M A. PEG-
stabilized carbodiimide crosslinked collagen-chitosan hydrogels for corneal tissue
engineering. Biomaterials. 2008;29:3960–3972.
242
Forbes JM, Cooper ME, Oldfield MD, Thomas MC. Role of advanced glycation
end products in diabetic nephropathy. J Am Soc Nephrol. 2003;14:254–258.
243
Yi H, Wu LQ, Bentley WE, Ghodssi R, Rubloff GW, Culver JN, Payne GF.
Biofabrication with chitosan. Biomacromolecules. 2005;6:2881–2894.
244
Albanna M Z, Akl T H B, Walters H L, Matthew H W T. Improving the
mechanical properties of chitosan-based heart valve scaffolds using chitosan fibers. J
Mechan Behav Biomed Mater. 2012;5:171–180.
245
Chatelet C, Damour O, Domard A. Influence of the degree of acetylation on some
biological properties of chitosan films. Biomaterials. 2011;22:261–268.
246
!
Madhavan K, Belchenko D, Motta A, Tan W. Evaluation of composition and
")(!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
crosslinking effects on collagen-based composite constructs. Acta Biomater.
2010;6:1413–1422.
247
Sionkowska A, Wisniewski M, Skopinska J, Kennedy CJ, Wess TJ. Molecular
interactions in collagen and chitosan blends. Biomaterials. 2004;25:795–801.
248
Han B, Jaurequi J, Tang BW, Nimni ME. Proanthocyanidin: A natural crosslinking
reagent for stabilizing collagen matrices. J Biomed Mater Res A. 2003;65:118–124.
249
Gniadecka M, Nielsen OF, Wessel S, Heidenheim M, Christensen DH, Wulf HC.
Melanoma diagnosis by Raman Spectroscopy and neural networks: Structure
alterations in proteins and lipids in intact cancer tissue. J Invest Dermatol.
1998;122:443–449.
250
Diem M, Romeo M, White SB, Miljkovic M´, Matthaus C. A decade of vibrational
micro-spectroscopy of human cells and tissue (1994–2004). Analyst. 2004;129:880–
885.
251
Tiwari A D, Mishra A K, Mishra S B, Arotiba O A, Mamba B B. Green synthesis
and stabilization of gold nanoparticles in chemically modified chitosan matrices. Int J
Biol Macromol. 2011;48:682–687.
252
Friess W, Lee G, Groves MJ. Insoluble collagen matrices for prolonged delivery of
proteins. Pharm Dev Technol. 1996;1:185–193.
253
Xu C, Wang Y. Cross-linked demineralized dentin maintains its mechanical
stability when challenged by bacterial collagenase. J Biomed Mater Res B: Appl
Biomater. 2011;96:242–248.
254
Wang Y, Spencer P. Continuing etching of an all-in-one adhesive in wet dentin
tubules. J Dent Res. 2005;84:350–354.
255
Sano H, Yoshikawa T, Pereira PNR et al. Long-term durability of dentin bonds
made with a self-etching primer, in vivo. J Dent Res. 1999;78:906–911.
!
"))!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
256
Sano H, Takatsu T, Ciucchi B. Tensile properties of resin-infiltrated demineralized
human dentin. J Dent Res. 1995;74:1093-1102.
257
De Munck J, Van Landuyt K, Peumans M et al. A critical review of the durability
of adhesion to tooth tissue: methods and results. J Dent Res. 2005;84:118–132.
258
Okada Y, Naka K, Kawamura K, Matsumoto T, Nakanishi I, et al. Localization of
matrix metalloproteinase 9 (92-kilodalton gelatinase/type IV collagenase = gelatinase
B) in osteoclasts: implications for bone resorption. Lab Invest. 1995;72:311-322.
259
Oster G, Holmstrom B. Riboflavin as an Electron Donor in Photochemical
Reactions. 1961; 83(8): 1867-1871.
260
Edwards AM, Silva E. Effect of visible light on selected enzymes, vitamins and
amino acids. J Photochem Photobiol. 2001;63:126–131.
#'" !Fernanda
Tranchesi S, Cecillia G, Paulo Eduardo C. Microtensile bond strength of
current dentin adhesives measured immediately and 24 hours after application. J Adh
Dent. 2005;7:297-302.!
262
Kohlhaas M, Spoerl E, Schilde T. Biomechanical evidence of the distribution of
cross-links in corneas treated with riboflavin and ultraviolet A light. J Cataract
Refract Surg. 2006;32:279–83.
263
Spoerl E, Mrochen M, Sliney D, Trokel S, et al. Safety of UVA-riboflavin cross-
linking of the cornea. Cornea. 2007;26:385–9.
#'% !Vesna
M, Pong P, Jan De M. Monomer-to-polymer conversion and micro-tensile
bond strength to dentine of experimental and commercial adhesives containing
diphenyl (2,4,6-trimethylbenzoyl) phosphine oxide or a camphorquinone/amine
photo-initiator system. J Dent. 2013;41:918-926.!
!
")*!
!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
265
Hashimoto M, Ohno H, Sano H. In vitro degradation of resin-dentin bonds
analyzed by microtensile bond test, scanning and transmission electron microscopy.
Biomaterials. 2003;24:3795-803.
266
Osorio R, Proenca JP, Erhardt MC. Resistance of ten contemporary adhesives to
resin–dentine bond degradation. J Dent. 2008;36:163–9.
267
Wollensak G, Spoerl E, Mazzotta C, Kalinski T, Sei S. Intralamellar cohesion after
corneal crosslinking using riboflavin and ultraviolet A light. Br J Ophthalmol.
2011;95:876-880.
268
Schmidt W. Light-induced redoxcycles of flavins in various alcohol/acetic acid
mixtures. Photochem Photobiol. 1982;36:699-703.
269
Huang R, Kim HJ, Min DB. Photosensitizing effect of riboflavin, lumiflavin, and
lumichrome on the generation of volatiles in soya milk. J Agric Food Chem. 2006;54:
2359-2364.
270
Fraser RD, MacRae TP, Suzuki E. Chain conformation in the collagen molecule. J
Mol Biol. 1979;129:463-481.
271
McCall AS, Kraft S, Edelbauser HF. Mechanisms of corneal tissue cross-linking in
response to treatment with topical riboflavin and long-wavelength ultraviolet radiation
(UVA). Invest Ophthalmol Vis Sci. 2010;51:129-138.
272
Changqi XU, Yong W. Cross-linked demineralized dentin maintains its
mechanical stability when challenged by bacterial collagenase. J Biomed Mater Res B
Appl Biomater. 2011;96:242-248.
273
Kohlhaas M, Spoerl E, Schilde T. Biomechanical evidence of the distribution of
cross-links in corneas treated with riboflavin and ultraviolet A light. J Catar Ref Sur.
2006;32:279–283.
274
!
Carrilho MRO, Geraldeli S, Tay FR, de Goes MF, Carvalho RM, Tjäderhane L,
"*+!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
Reis AF, Hebling J, Mazzoni A, Breschi L, Pashley DH. In vivo preservation of the
hybrid layer by chlorhexidine. J Dent Res. 2007;86:529-533.
275
Cadenaro M, Breschi L, Rueggeberg FA, Suchko M, Grodin E, Agee KA, Lenarda
RD, Tay FR, Pashley DH. Effects of residual ethanol on the rate and degree of
conversion of five experimental resins. Dent Mater. 2009;25:621-628.
!
"*"!
[...]... unwinding of the triple helix in the alpha chain (Nagase H, Fushimi K Elucidating the function of noncatalytic domains of collagenases and aggrecanases Connective Tissue Research 2008;16:9:74) ! #%! Fig 19 Obtained Raman spectrum of the (a) control demineralized specimen; (b) Adper TM Single Bond; (c) and chitosan in the region of 700-1700 cm-1 (Daood et al; Effect of chitosan/ riboflavin modification on resin/ dentin. .. micrograph of the resin- dentin interface bonded with 3M Single Bond ESPE (1500x) (a) dental composite; (b) adhesive bond; (c) hybrid layer; (d) resin tags ! ##! Fig 17 Scanning electron microscopic image of the resin- dentin interface bonded with 3M Single Bond ESPE A uniform hybrid layer (HL) formation can be seen with visible resin tag (RT) formation ! #$! Fig 18 Collagenolysis in the presence of collagenase... resin/ dentin interface: Spectroscopic and microscopic investigations) ! #&! Fig 20 Schematic representation of the specimen preparation for the micro-tensile bond strength (!TBS) test and SEM analysis (A) Removal of occlusal surface to expose the flat dentin surface; (B) preparation of the dentin surface to receive adhesive with or without RF; (C) cutting of resin- dentin beams from the center of bonded... (arrow) on top of the resin tags and a relatively thicker and more textured hybrid layer; HL, hybrid layer; RT, resin tags (Daood et al; Effect of chitosan/ riboflavin modification on resin/ dentin interface: Spectroscopic and microscopic investigations) ! $+! Fig 25 Raman spectra of control, 0.1%RF and Ch/RF (1:4 and 1:1) crosslinked dentin specimens in the spectral range of 900–1700 cm-1 The peak assignments... can be observed in the specimen interface treated with 0.1%RF crosslinking prior to dentin bonding agent application The resin cement penetrated deeply and many long resin tags were observed at the demineralized interface (C) A funnelshaped configuration of the resin tags also seen at the base of Ch/RF 1:4-treated specimens The resin tags exhibited a slightly rough texture (D) The resin tags in Ch/RF... illustrative area of the dentin surface of (A) control; bio-modified with (B) 0.1%RF; (C) Ch/RF 1:4 and (D) Ch/RF 1:1 specimens The 0.1%RF and Ch/RF 1:4 specimens show open dentinal tubules with intact collagen fibers whereas the Ch/RF 1:1 specimens exhibit a discontinuous structure The dentin of all specimens were conditioned for 15 s with 37% phosphoric acid (Daood et al; Effect of chitosan/ riboflavin modification. .. specimen, and (C) resin impregnated dentin recorded in the region between 800-1800 cm-1 The P-O bond at 960 cm- 1of the mineral component is well represented for demineralized and resin impregnated dentin The peaks at 1667 cm-1 and 1246 cm-1 are associated with organic components for dentin collagen ! #)! Fig 23 Representative SEM micrographs of the etched dentin resulting from different bio -modification. .. chitosan/ riboflavin modification on resin/ dentin interface: Spectroscopic and microscopic investigations] ! $"! Fig 26 Raman line map (A) and images of the spectrum at the 4 !m (B) and 8 lm (C) crosslinked resin/ dentin interfaces Intensities at 960 cm and 1450 cm-1 for all other specimens are identified in the line map The Raman images indicate the positions of spectra in the region of interest The spectrum is characterized... Table II The C-H alkyl groups also appeared in the collagen spectrum in dentinal substrate with other peak areas of CAC bond, Amide I and Amide III Schematic representation of CH3 inplane bending also shown (a) control (b) 0.1%RF (c) Ch/RF 1:4 (d) Ch/RF 1:1 [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com ; Daood et al; Effect of chitosan/ riboflavin modification. .. figure can be viewed in the online issue, which is available at wileyonlinelibrary.com Daood et al; Effect of chitosan/ riboflavin modification on resin/ dentin interface: Spectroscopic and microscopic investigations] ! $#! Fig 27 SEM images of the resin- dentin interface after 24 h storage in artificial saliva Well-defined uniform hybrid layer (white arrows) and with well-formed branched resin tags could be ... spectrum of the (a) control demineralized specimen; (b) Adper TM Single Bond; (c) and chitosan in the region of 700-1700 cm-1 (Daood et al; Effect of chitosan/ riboflavin modification on resin/ dentin. .. without RF; (C) cutting of resin- dentin beams from the center of bonded specimen; (D) attachment of the single resin- dentin beam for immediate !TBS testing; (E) storage of resin- dentin beams in artificial... (arrow) on top of the resin tags and a relatively thicker and more textured hybrid layer; HL, hybrid layer; RT, resin tags (Daood et al; Effect of chitosan/ riboflavin modification on resin/ dentin interface: