SEM Investigation of the Variability of Enamel

Surfaces After Simulated Clinical Acid Etching

for Pit and Fissure Sealants
G. W. MARSHALL, L. M. OLSON, and C. V. LEE
Department of Biological Materials, Northwestern University Dental School,
Chicago, Illinois 60611, USA

Etching preferentially occu-s on cuspal inclines and on premolars vs molars. After

longer times, etching occurs near the pits
and fissures. Enamel rods etch at cores and
prism borders simultaneously in the same
area. Longer etching times produce more
enamel surface relief, permitting better
sealant adaption to the enamel.
Pit and fissure caries is a very destructive
and rapidly advancing disease of the teeth.
Several methods have been tried in an attempt to stop or retard this process. Among
the more well-known and possibly more effective methods are fluoride treatments, good
oral hygiene, and the relatively new method
of pit and fissure sealants.
The main objective of pit and fissure
sealants is to prevent bacterial colonization
and plaque formation which are implicated
in the cariogenesis of developmental pits,
fissures, and grooves. A tight seal denies
plaque colonization and physically prevents
accumulation of food debris both of which
support the bacteria at these preferred sites.
One of the major problems in pit and fissure
treatment is to effectively bond the sealant
to the enamel surface.
Buonocorel first used acidic conditioning
treatments of enamel surfaces to increase
the adhesion of acrylic to enamel. Acid
etching introduces microirregularities into
the enamel structure that are penetrated
by the fluid sealant and then mechanically
This article was awarded first place at the Annual Research Award Competition, Chicago Section, AADR,
1974.
Received for publication June 24, 1974.
Accepted for publication June 27, 1975.

1222

locked into place on polymerization of the

sealant. Clinical reports2,3 have shown that
etching with 50% phosphoric acid solutions
can be used with BIS-GMA methyl methacrylate polymer sealants to reduce pit and
fissure caries in permanent and deciduous
teeth. Ohsawa4 found that the improvement
in adhesion after etching was not a function
of the amount of decalcification that took
place but was dependent on the surface
characteristics created by the acid etching.
Gwinnett and Buonocore5 reported that acid
etching was limited to the inclined planes
of the cusps and did not reach into the pits
and fissures of the tooth. This probably has
little effect on the initial adhesion of the
sealant, but with sealant attrition on the
cusps because of masticatory wear, it could
result in loss of sealant.
Organic and mineral acids reportedly dissolve enamel prism necks and cores, whereas
ethylenediaminetetraacetic acid initially demineralizes the enamel rod periphery.6.'1
Etching patterns parallel to the major axis
of the enamel rods are usually described as
fish scales or keyholes, but a variety of paterns have been reported.12(PP 187,188)
Gwinnett13 described the structure of a
prismless layer on the surface of permanent
teeth. This layer may be more resistant to
dissolution than the underlying prismatic
enamel. The term "prismless enamel" is
used here to describe portions of the enamel
that lack the prismatic pattern.
Lee, Phillips, and Schwartz14 found that
etching enamel with 50% phosphoric acid
produced a mechanical strength with the
polymerized sealant that exceeded the
strength of the sealing material. Martinezl5
found that several commercial sealants

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ACID ETCHING FOR PIT & FISSURE SEALANT

1223

slhowed good in vitro sealability after toothbrushing and temperature cycling.

The present investigation attempted to
clharacterize the variety of acid-etched patterns producedl during simulated clinical
etchiing witlh 50% plhosphoric acid containing 7% dissolved zinc oxidea as a function
of etchiing time on the enamel surfaces of
premolars and molars. The resulting patterrns were related to the adaption of a commercial pit anid fissure sealant when applied
to the treated enamel suLrface.
Materials and Methods

Noncarious, extracted human molars and

piemolais were selected for acid etclhling and
sealant studies uIsing a scanning election microscope (SE NI) 1, 'Ilhe teetlh were scr tbbed
withi watei app))lied witlh a cotton pledget
and were stored in commercial grade alcohol
after extraction. They were allowed to
lench (Iry 24 liours before SET\I examinationl.
No p)rophylaxis treatments were used. Tihe
uinetchiedI enamel surfaces were examined for
later comparison witlh the etclhed surfaces.
This initial exposure to the SEMN atmosphiere
did not produce any apparent structuiral
modification since the same etching patterns
were observed even if this step was eliminated.
After study of the uinetclhed surfaces, the
teethl were etclhed Lusing the recommended
clinical application teclhniquies. The occlusal
suirfaces of the teeth were etched witlh buffered 50%) phiosplhoric acid applied with a
cotton swal) for time periods of 30, 60, 75,
90, 120, or 240 seconds. The teeth were
then rinsed witlh tap water, air dried, and
reexamined under tlhe SEM. Th-e occlusal
surfaces of representative teeth etched for
each of these time periods were coated with a
commercial sealante and polymerized with
tultraviolet liglht foi 60 seconds. Several of
these teetlh were sectioned to yield mesiodistal or buccolingual sections through the
pits or fisstrres. These sections were later
uLsed to study the adaptation of the sealant
at the enamel-sealant interface.
A thin layer (200 A) of gold was vacuum
evaporated onto the sealant suLrface after
the sealing or sectioning procedures. The
gold laver was necessary to prevent charge
nNova-Seal con(litioning agent, L. D. Caulk Co., Mil-

ford, I)el.

b Stereoscan S-4 scanninig electron microscope, Kent

Cambridge Scientific Instruments, Ltd., Cambridge, Eng.
e Nuva-Seal, L. D. Caulk Co., Milford, Del.

FIC 1.-SEM photomicrograph of unetched

enamel. Upper portion shows relationship of
cusps and central groove of molar. Groove contains debris. Lower portion shows typical appearanace of unetchedt enamel on cuspal inclines
at right on upper portion at highi magnification.
Relatively smooth enamel is visible with numerous minor depressions in enamel before etchi-

inig. (. = pm)
buildup on the electrically insulating sealant

dIuring electron bombardment. Each of the

sealed or sealed-and-sectioned teeth was examined withl the SEM for surface defects in
the sealant an(d for sealant bonding at the
enamel-sealant interface. The SEM was operated at ain accelerating potential of 20 kv
witlh a 200-micrometer (btm) diameter final
a-)erture for all samples in this investigation.

FIG 2.-Photomicrograph of enamel on cuspal

incline midway between cusp tip and groove
after 30-second etching treatment with 50%
phosphoric acid. Enamel was lightly etched on
surfaces of premolars. Cores of enamel rods
were preferentially dissolved by etchant. (s=

pm)

1224

MARSHALL, OLSON, AND LEE

J Dent Res November-December 1975

FIw 3,-Photormicrographs showv effect of 60-seconid etchinig trcatmenlt onl occlusal su-face
of priemolar. Enamel on cuspal inclines was fairly well etched (left), but only light, nonuLniform etchiing occurred near the fossa (right) (,u =-1m)

Results

B3efore acidl etclinrg, the enamel surfaces

weie relatively smooth over the entire inner
aspect of tlhe cusps. Frequently, debris could
he seen in thie cential groove, pits, and fissures of the teeth. Figture I slows a typical
enamel surface from a molar before etclhing.
The top portion of Figure I is a low-magnificattion, SENI photomicrograph sthowing the
general culsp aniatomy and debris in the
groove. The lower portion of this figure
slhows the fairly smooth aspect of the enamel
on tile cLuspal inclines of tlhe same tootlh. A
numher of minior depressions can be seen
b)efore tde etclhing treatment. After 30-second
etcliing times withi phosphoric acid, the cuspal inclinies of molairs and premolars showed
signs of liglht etclhing (Fig 2). The begin-

ning of etched prism patterns is clearly

seen in this figure and tlhe etcliing attack
was concentrated in the core region of the
enamel irods. T1he etchlinig pattern was uncxen and fui -thier dlown tlhe ctusps, little evidleuce of etcling could le found. Figure 3
slhows the clharacteristic etching patterns obsei-ved on- prernolars after etclhing for 60 secondcls. EI'lie acid dissolution of enamel higher
on- the culsps in Figure 3, left appeared to he
deeper and more uniform than the etching
near the developmental pit in Figure 3,
i-ighlt. The 60-secondI etching patterns on
molars were less well developedl (Fig 4).
Near tlle cu1sp ti p, Figure 4, left, etclhing
effects were evideent, but a prismless layer at
the lower right apparently interfered witl
the continuLouis development of enamel rod

FIG 4.-Etched enamel patterns are shown after 60-second etching treatmelnt on molar. Near
cusp tip (left), enamel prism patterns can be seen in upper left, but much of surface was
covered with prismless layer (lower right) Near central groove (right) , there wvas little
apparent effect of etching attack on enamel. (,u=-um)

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ACID ETCHING FOR PIT & FISSURE SEALANT

1225

end patterns as seen at the upper left. Little

evidence of etching near the pit was seen in
the molars (Fig 4, righit).
Figure 5 sihows an unusual result of etching on the uipper part of the cuspal inclines
on a premolar etched for 75 seconds. A high
(legree of surface roughness and patchies of
well-developed enamel rod patterns were
produced. In the center portion of Figure
5, a region of exposecd and etched dentin is
apparent. (Note the dark holes whlich are
the etchled dentinal tubules.) This region
could hlave been exposed because of a localized legioni of thin enamel that was etched
atwa. 1 hle dentin appears dark beca use it is
mulch less hiiglhly mineralized and tlhetefore
emits fewer secondary electrons per u-nit
area.

Characteristic etching patterns on tlhe cui-s

pal inclines of a1 premolar after 75 seconids

of etcliing are slhown in Figutre 6. XVelldeveloped enamel rods are seen interspersed
witl)poorly etclhed areas in Figniie 6, top.
Figuire 6, hottom is a hiigler-magnification
view of Figure 6, top and slhows that etching
of the enamel rod periphery predominated
in this portioni of the sample. This gave a
typical fishi scale appearance to the enamel
prisms. Etcling of the occlusal su-rfaces of
molars resuLlted in less well-developed rod
pattterns (Fig 7) A band of well-etched
enamel rods are seeni in Figuire 7, top, sandwiched between prismless enamel regions
that resisted etching. Figutle 7, bottom slIows

FiG 5.-Cuspal enamel from premolar etched

75 seconds. General roughening of surface ancd
ai-eas of enamel rodIs are apparent. In center
of micrograph, region of exposed andI etclhedI
dlentin canl be seen. Note dentinal tubules at
center of region. (u =gim)

EF.; 6.-Typical etching patterns on cuspal inclines of premolar after 75-second etching treatment. Urxeven or mottled appearance of pattern
is apparent (top). At higher magnification,
tendency for acid attack on enamel prism periphery is shown (bottom). (L -gim)
only the faint beginnings of etched enamel
patterns throughl the prismless layer near the
central fossa of the same tooth. It should
also be noted that in Figure 7, bottom etching appeais to occur first at the prism cores.
Etching time periods of 90 seconds produced well-developed prism patterns on the
cuspal inclines as shown in Figure 8, left,
witlh patclhes of a prismless layer still evident.
Etching patterns extended downward along
the cusps almost to the periphery of the developmental pits as seen in Figure 8, right.
T1he prism patternis were more uniform
on tlhe premolars etchled for 120 seconds
(Fig 9) than the piism patterns on molars
etclhed for 240 seconds. After the 240-second

1226

MARSHALL, OLSON, AND LEE

J Dent Res November-December 1975

FIc 9.-Well-developed enamel rod pattern

on cuspal surface of premolar after 120-secondl

etchiing treatment. (,

,um)

etclinlg periods, the enamel prism patterns

on the inclines of premolars were well developed, and a typical mosaic pattern was
evident as seen in Figure 10, left. Figure 10,
right is a higher magnification of the same
structure and shows the common observation
that some patches of rods were preferentially
attacked at rod peripheries wlhereas otlher

patches suffered preferential

FIG 7.-Etching patterns on molar occlusal

after 75-second etching tieatment. Prismless layer covered much of surface on cuspal incline and interfered with etchling of enamel rods
(top) Etched rods are seen at center with etching at rodI bordeis. Very light etchiing wvas seen
near fossa (bottom) . Acid attack through prismless layer occurred at rod cores. (,= ,um)

suirface

dissolution of

the rod centers and tails. Figure 11 shows

tlhat etclhing patterns were developed at the
eIge but not on the steeply inclined walls
of developmental grooves after the 240-seco(id etcliing treatment.
\pplication and polymerization of the
sealant resulted in a smooth surface covering
the desvelopmental pits and fissures. The
sealant surfaces did contain numerous pores

FIG 8.-Etched enamel on premolar after 90-second etching treatment. Left, well-etched
enamel rods interspersed with remnants of prismless layer on cuspal incline. Right, lowermagnification view shows approach of etching patterns to edge of pit. (gi = fm)

ACID ETCHING FOR PIT Qr FISSURE SEALANT

Vol 54 No. 6

1227

FIG 10.-Enamel etching patterns on cuspal incline of premolar after 240-second etching treatment. Ieft, lower-magnification view shows mosaic or patchy appearance of etched enamel.
Rig/lt, at lhigher magnification, patches of rods can be seen that etched initially at prism borders
and other patches can be seen etched preferentially at prism cores. Region of unetched or
prismless enamel appears to separate most similarly etched patches. (j = xm)

over a wide-size range. Figure 12 shows a

typical occlusal surface view of the sealant
in a developmental pit of a molar. Large
pores of more than 50-1tm diameter are evident in the surface. Smaller pores are also
evident, as shown at the arrow.
Figures 13 to 15 show the adaptation of
the sealant to the etclhed enamel in sections
througli the fis.sure. The relatively poor
adaptation of the sealant at the enamel walls
after etcling for 30 seconds witlh phosphoric
aicidl can be seen in Figure 13. Figtire 14 is
a photomicrograplh of a small portion of the
eniamel-sealant interface obtained from a section tlhrough a fissuire with use of a 60-second
etching treatment. The adaptation of the
sealant is much better than after the 30second etclhing treatment. A similar view of

the excellent adaptation of the sealant to

the etched enamel after a 240-second etclhing
treatment is shown in Figure 15.

Discussion
The acidic conditioning treatments using
50% phosphoric acid increased the enamel

FIG 11.-SEM photomicrographs of etching

patterns

near central

groove after 240-second

etch of premolar. Enamel rod patterns are seen

interspersed with clumps of remaining prismless layer up to steep inclines of walls (top)
Higher magnification view at groove wall shows
border between well-etched and poorly etched
enamel at gx-oove (bottom). (g in)

4.

8
I

1228

MARSHALL, OLSON, AND LEE

FiG 12.-SEM view of sealant on occlusal surface of molar. Varying diameter bubbles were
observed as defect of application. They ranged
in size from relatively large ones shown at uspper
left to very small micropores (arrow)
surface area and produced a variety of etclhing patterns into whlichi the sealant could key
and form a mechanical bond. Slhorter etching times resulted in etclhed enamel rod
patterns on the cuspal inclines but did not
etclh the groove region significantly as shown
in Figures 2 to 4. Generally, increased etchiing time resulted in the development of
etching patterns over a greater portion of
the enamel surface. Longer etclhes produced
prism patterns that reachedl the periphiery
of the steep inclines of tlevelopmental
grooves or pits and fissures. This effect can

FiG. 13.-Enamel sealant interface in section

through fissure after 30-second etch and polymerization of sealant. Bubble in sealant penetrated halfway through sealant at upper middle.
Sealant was poorly adapted at interface. (,u=

gm)

J Dent Res November-December 1975

FIG 14.-High-magnification photomicrograph

of enam-lel-sealant interface in section throtigih
fissure after 60-second etching treatment. Enamel is at left and sealant at right. Tighter
jiuinction between enamel and scalant is evident
as compared with Figure 13. (,A=im)

he seen by comparing Figure 8, right and

Figuire 11. to1). The grooves, pits, and fissures were Inot etcled, possibly as a result of
a film of organic material an(d debris coverinig these areas. This film apparently offers
a bai-rier to the general etching attack on
the enamel. However, it is to he expected
that localized areas of the pits and fissures
couldlibe attackedl in view of the prevalence
of pit and fissure caries. Suclh localized attack in the pits would not le beneficial for
aclhieving sealant adhesion.

FIc. 15.-SEM view of very tight seal obtained in section after etching of enamel for
240 seconds. Sealant-enamel interface was so
well adapted that it is difficult to locate boundary. Sealant at left, enamel at right. ( =- inm)

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ACID ETCHING FOR PIT & FISSURE SEALANT

A high degree of variation was noted in

the etching patterns from tooth to tooth and
in different parts of the same tooth given the
same "clinical" etching procedure. Less welldeveloped etching patterns were found on
molars than on premolars given the same
treatment, for example, compare Figures 7
and 8. Local anatomy would be expected to
have an effect on the etching patterns' characteristics. Teeth with less steeply inclined
developmental grooves gave etching patterns at inferior positions on the walls as
compared with teeth with steeper inclines.
Etching patterns did not develop within the
grooves or fissures for any of the samples
studied. This is apparent in Figures 4, right,
7., bottom, and 11, and is in agreement with
the observations of Gwinnett and Buonocore.5
The nature of the acid etching attack on
the enamel surfaces studied can be characterized as patchy. This phenomenon had
two major manifestations. First, certain portions of the enamel surface appeared to have
a thicker, denser, or more tightly adhering
prismless layer that impeded dissolution of
the underlying enamel prisms. This layer
was more evident near the pits than on the
cuspal inclines as shown in Figure 2 and was
also more evident in molars than premolars
as seen by comparison of Figures 7 and 8.
Occasional patches of this layer could be
found on various surfaces of all teeth, even
after the longest etching times used. The
etched surface of Figure 11, top is covered
with clumps of this layer that are surrounded
by well-etched enamel rods. Increased etching times tended to promote a more evenly
roughened surface with fewer patches of
the prismless layer. These more uniformly
etched and cleaned surfaces are required for
good adhesion of the sealant.
The second form of the patchy nature of
the etched surfaces is characterized in Figure
10, although it was a commonly observed
phenomenon. Portions of the enamel rods
were preferentially etched at prism cores and
necks in some areas, whereas in other adjacent areas the periphery of the enamel rods
were etched away. Both forms of etching
dissolution were seen throughout this investigation. Generally, a small portion of
closely packed and similarly etched rods
were bounded by a narrow, prismless region.
This gives the impression of a structure similar to grains in metallographic specimens in

1229

which regions of high order etch differently

than their boundaries. These observations
suggest that patches of enamel rods tend to
align and are separated by regions of mismatch to accommodate the curved surface of
the tooth. This structure would be in contrast to a structure composed of enamel rods
with more or less uniform mismatch to accommodate the variations in the surface topography of the teeth. Transmission electron micrographs suggest that some orientational mismatches do occur between adjacent
enamel rods.12 (p182) Continued investigation
would be necessary to determine how the
relative rod orientations change with small
radii of curvature at the tooth surfaces.
A second explanation of the mosaic appearance of the etched enamel could depend
on the adhesion of the prismless layer discussed previously. Each patch of similarly
etched enamel rods could be a result of being covered with a prismless layer that was
dissolved away at one time. Thus, regions
such as that at the center of Figure 7, top
would be etched similarly and the areas on
either side that are still covered with the
protective layer might etch differently. It
should be noted that many of the boundaries
surrounding similarly etched rods in Figure
10 appear similar to the layer covering the
major portion of the surface in Figure 7, top.
Neither of these hypotheses for the mosaic appearance of the etched enamel accounts for the differences in etching attack
of enamel rods observed in adjacent areas
of the samples. Previous work reported
that acids preferentially dissolve the prism
cores.5,12 (p182) The present investigation
demonstrates that after the 60-second etching time, attack at prism borders is as likely
as attack at the core. It appears that the
first layers etched may be characterized by
preferential attack at prism cores as shown
in Figures 2, 3, right, and 7, bottom. After
etching through the prismless layer and an
initial layer of prismatic enamel further dissolution can occur at prism borders or cores
with apparent equal probability. This observation is in conflict with most published
work, although etching at peripheries and
cores has been reported.16 This suggests that
in deeper enamel, adjacent groups of prisms
exist in different states of mineralization, or
that particular groups of prisms have a
higher affinity for the prismless layer. A definitive explanation for this difference in sol-

1230

MARSHALL, OLSON, AND LEE

ubility will depend on a more complete understanding of the dissolution mechanism.

Acid etching of enamel is the most popular and presently most effective way to obtain adhesion of pit and fissure sealants to
enamel. It is axiomatic that such treatments
should produce a sealed surface for as long
as possible. Figure 12 shows the range of
sizes of bubble defects that developed during polymerization of the sealant. The micropores are of doubtful clinical significance,
but the larger defects represent severely
weakened areas that would break down under occlusal forces. Such sealant loss undoubtedly would shorten the protective lifetime of the sealant since much less sealant
would remain to be abraded.
Figures 13, 14, and 15 show the increased
adaption of the sealer to the etched enamel
with increased etching time. Closer adaption of the sealant to the enamel is interpreted as both increased marginal sealability
and adhesion. These results suggest that
there is a significant advantage in having a
clean, debrisfree, and uniformly roughened
surface for good sealability and that this condition can be partially achieved by longer
etching times, for example, up to four minutes for molars. Furthermore, the tight adaption approached the steep inclines of the
pits, fissures, and grooves because of the
more uniformly etched enamel in this region
after extended etching times. This should
extend the effective lifetime of the sealant
because the fissure would continue to be
sealed even after the sealant on the cuspal
inclines was partially abraded.
The objectives of using longer etching
times included enhanced removal of the
prismless layer and increased etching close
to the steep walls of the pits and fissures.
These factors could outweigh the importance
of longer etching to achieve increased depth
in the microirregularities associated with the
prisms, although such changes have been
reported by Silverstone.17 If other methods
can be developed to efficiently remove the
prismless layer, then shorter etching times
might be used to obtain more uniform etching patterns and good adaption of sealant to
the etched enamel surface.

Conclusions
The present investigation of etching patterns on enamel obtained during simulated
clinical etching experiments in vitro was

J Dent Res November-December 1975

undertaken to determine differences in the
etching patterns that would be encountered
in the clinical situation and might affect the
adhesion of pit and fissure sealants.
In agreement with previous studies, etching with 50% phosphloric acid increased the
enamel surface area and produced etched
enamel prisms. Deep pits fissures, and
grooves are not etched, and preferential
etclhing occurred on the cuspal inclines.
In contrast with other work, etching occurred both at prism borders as well as
prism cores in subsurface enamel. Neithei
form appears to be dominant. The initial
layer of enamel appeared to be preferentially attacked at the core of the enamel
prisms.
Significant differences in the etching patterns occurred on various portions of the
occlusal surface of individual teeth and differences also occurred between molars and
premolars. Portions of a single tooth appeared to have a more adherent, prismless
layer near the pits, fissures, and developmental grooves that was resistant to etching. All molars were more resistant to etching as compared to premolars.
To obtain a debrisfree, well-etched,
enamel structure, extended etching times
were required. Better adaption of sealant to
etched enamel walls occurred with longer
etching times and etching was more uniform
near the pits and grooves. All the evidence
gained in this investigation suggests that
longer than normal etching times would
produce a more adherent and longer lasting
sealant, especially in preventive treatment
of molars with pit and fissure sealants.

References
1. BUONOCORE, M.G.: A Simple Method of Increasing the Adhesion of Acrylic Filling Materials to Enamel Surfaces, J Dent Res 34:

849-853, 1955.
2. BUONOCORE, M.G.: Adhesive Sealing of Pits
and Fissures for Caries Prevention with Use
of Ultraviolet Light, JADA 80: 324-328, 1970.
3. BUONOCORE, M.G.: Caries Prevention in Pits
and Fissures Sealed with an Adhesive Resin
Polymerized by Ultraviolet Light; A Two
Year Study of a Single Adhesive Application,
JADA 82: 1090-1093, 1971.
4. OHSAWA, I.: Studies on the Solubility and
Adhesion of the Enamel in Pre-Treatment
for Caries Preventive Sealing, Bull Tokyo
Dental Coll 13: 65-82, 1972.

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ACID ETCHING FOR PIT & FISSURE SEALANT

5. GwINNErr, A.J., and BUONOCORE, M.G.: A

Scanning Electron Microscope Study of Pit
and Fissure Surfaces Conditioned for Adhesive Sealing, Arch Oral Biol 17:415-423,
1972.
6. POOLE, D.F.G., and JOHNSON, N.W.: The
Effects of Different Remineralizing Agents
on Human Enamel Surfaces Studied by
Scanning Electron Microscopy, Arch Oral
Biol 12:1621-1634, 1967.
7. HOFFMAN, S.; McEwAN, W.S.; and DREW,
C.M.: Scanning Electron Microscope Studies
of EDTA-Treated Enamel, J Dent Res 48:
1234-1242, 1969.
8. ScoTT, D.B.; SIMMELINK, J.W.; and NYGAARD,
V.: Structural Aspects of Dental Caries, J
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9. SHARPE, A.N.: Influence of the Crystal
Orientation on Human Enamel and Its Reactivity to Acid as Shown by High Resolution Micro-Radiography, Arch Oral Biol 12:
583-591, 1967.
10. SWANCAR, J.R.; Sco rr, D.B.; and NJEMIRovSKIJ, Z.: Studies of the Structure of Human
Enamel by the Replica Method, J Dent Res
49: 1025-1033, 1970.
11. NICHOL, T.; JUDD, G.; and ANSELL, G.W.: A

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Two Stage Model for Human Enamel Demineralization as Determined by Scanning

Electron Microscope Analysis, J Dent Res 52:
487-493, 1973.
12. Scorr, J.J., and SYMONs, N.B.B.: Introduction
to Dental Anatomy, 6th ed, Baltimore:
Williams & Wilkins Co., 1971, pp 182, 187188.
13. GWINNETr, A.J.: The Ultrastructure of the
"Prismless" Enamel of Permanent Human
Teeth, Arch Oral Biol 12: 381-387, 1967.
14. LEE, B.D.; PHILLIPs, R.W.; and SWARTZ, M.L.:
The Influence of Phosphoric Acid Etchings
on the Retention of Acrylic Resin to Bovine
Enamel, JADA 82:1381-1386, 1971.
15. MARTINEZ, C.R.: The Effect of Three Variables on the Seal of Three Commercial Pit
and Fissure Sealants, MS thesis, Northwestern University, 1973.
16. GwINNETT, A.J.; BUONOCORE, M.G.; and
SHEYKHOLESLAM, A.: Effect of Fluoride on
Etched Human and Bovine Tooth Enamel
Surfaces as Demonstrated by Scanning
Electron Microscopy, Arch Oral Biol 17:

271-278, 1972.
17. SILVERSTONE, L.M.: Fissure Sealants/Laboratory Studies, Caries Res 8: 2-26, 1974.