Caries Res.

9: 373-387 (1975)

Variation in the Pattern of Acid Etching of Human Dental

Enamel Examined by Scanning Electron Microscopy

L. M. SiLVERsroNE, C. A. Saxton, I. L. D ogon and O. F ejerskov

The London Hospital Medical College, Dental School, London.

Gibbs Dental Research, Unilever Ltd., Isleworth Laboratory, Isleworth, England
Harvard School of Dental Medicine, Boston, Mass.,
and The Royal Dental College, Aarhus

Key Words. Acid etching • Enamel • Etching patterns • Prisms

Abstract. This study has shown that the exposure of human dental enamel to
acid solutions in vitro produces three basic etching patterns. In the most common,
called type 1 etching pattern, prism core material was preferentially removed leav­
ing the prism peripheres relatively intact. In the second, type 2 etching pattern, the
reverse pattern was observed. The peripheral regions of prisms were removed
preferentially, leaving prism cores remaining relatively unaffected. In the type 3
etching pattern, there was a more random pattern, areas of which corresponded to
types 1 and 2 damage together with regions in which the pattern of etching could
not be related to prism morphology. These findings differ from previous studies in
which the type I pattern was ascribed to acid action and type 2 etching pattern to
attack by chelators. The results therefore suggest that there is no one specific
etching pattern produced in human dental enamel by the action of acid solutions.
Such differences produced by acids are difficult to explain on the basis of varia­
tion in chemical composition, and crystallite orientation. This further highlights
the variation in structure that can occur in enamel not only from tooth to tooth,
or surface to surface, but also from site to site on a single tooth surface.

In a previous publication P oole and J ohnson (1967] described dif­

ferences in etching patterns produced by the action of dilute mineral ac­
ids, dilute organic acids at pH 4.0, and ethylene diamine tetra-acetic
acid (EDTA) at pH 7.0, on the surface of human dental enamel seen
with the scanning electron microscope. When the natural surface of a
tooth, or a surface prepared transverse to prism direction, was exposed
to acid, a characteristic honeycomb pattern was produced. This was in­
terpreted as resulting from preferential loss of prism centres. Mineral
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374 Silverstone /S axton D ogon/F ejerskov

and organic acids apparently had similar effects. The EDTA solution al­
ways produced what appeared to be the opposite effect. Cylinders of
material were left projecting towards the original surface, separated by
gaps produced by an apparent preferential dissolution of the peripheral
parts of the prism. These findings were first recorded by M uller and
Schait [1957] using replica techniques and have subsequently been con­
firmed by H offman et al. [1968, 1969a, b] using the scanning electron
microscope. In a further study, J ohnson et al. [1971] found, once again,
that acid solutions preferentially removed prism centres whilst EDTA, at
neutral pH, preferentially removed prism peripheries. However, short
exposures to dilute acids produced narrow clefts at prism peripheries as
well as shallow concavities in the body of the prism. G winnett [1971)
examined enamel after exposure to acid solutions and found different
surface characteristics in experimental regions, none of which were ex­
clusive to any one particular acid. The most common finding was that in
which prism core material was preferentially removed. The reverse of
this pattern was observed in some cases after acid action. G winnett el
al. [1972] and Sheykholeslam and B uonocore [1972] also showed
etching patterns in which there was either a preferential loss of prism
core material or prism peripheries, both patterns being produced by the
one acid solution. Thus, the pattern of etching in which prism cores are
left relatively intact with a preferential removal of the peripheral materi­
al does not necessarily appear to be confined to attack on enamel by
chelating agents.
This study was carried out to see if there is one specific etching pat­
tern produced by exposure of surfaces of human dental enamel to acid
solutions.

Materials and Methods

Sixty sound human premolar teeth, extracted for orthodontic purposes, were
employed in this study. Small window regions of enamel were prepared on either
smooth surfaces or cuspal slopes by painting the rest of the tooth with varnish
and then they were exposed to one particular acid solution. The acid solutions
employed were unbuffered phosphoric acid in the concentration range 20-70°/o,
and buffered solutions of lactic acid at 1, 0.1 and 0.001 n at pH 4.5. Exposure
times varied from 1 to 10 min when employing phosphoric acid solutions and 1-5
days with lactic acid solutions. None of the solutions was stirred during an expo­
sure. In this manner, damage to the superficial layers of enamel was observed in
the group of experiments employing phosphoric acid whilst with lactic acid, etch­
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 Variation in the Pattern of Acid Etching 375

ing patterns were observed in the inner regions of enamel. After exposure, speci­
mens were thoroughly washed with distilled water and dried with compressed air.
Specimens were then glued to aluminium stubs with the exposure window upper­
most and vacuum-coated with a layer of carbon followed by silver or gold, for ex­
amination in the scanning electron microscope. The total thickness of the coating
was estimated at approximately 50 nm.
After exposure to phosphoric acid, a number of specimens were coated with a
fissure sealant. Two materials were used, both of which were polymerized by ul­
tra-violet light (Nuva-Seal® and TP 226) [Silverstone, 1974], After application
of the sealant and its subsequent polymerization, specimens were demineralized in
12-percent hydrochloric acid for 5 h. In this manner, all of the dental tissues were
removed, leaving just the plastic-type sealant remaining. The fitting surface of the
sealant thus acted as a replica of the previously etched enamel surface. These
specimens were also vacuum-coated for examination in the scanning electron mi­
croscope.

Results
Phosphoric Acid
When using phosphoric acid solutions within the short exposure
times employed in this study, three patterns of enamel surface etching
were seen.
In the first, called type 1 etching pattern, there was a generalized
roughening of the enamel surface, but with a distinct pattern showing
hollowing of prism centres with relatively intact peripheral regions
(fig. 1). The average diameter of the hollowed regions was 3 ,«m. This
was found to be the most common of the three patterns observed.
In the second, or type 2 etching pattern, prism peripheries appeared
to be removed, or heavily damaged. Therefore, the prism cores were left
projecting towards the original enamel surface (fig. 2). This apparent
damage of the peripheral regions of the prisms was seen to extend along
the length of the prism, thus aiding in delineating individual prisms
(fig. 2). When viewed from the original surface (fig. 3), separate bundles
or columns of material are seen, the gaps separating them corresponding
to the peripheral regions of the prisms. Thus, this type 2 etching pattern
was the reverse of the honeycomb pattern of type 1 damage, and both
patterns were produced by exposure to a similar solution of phosphoric
acid for an identical exposure time.
Some etched regions showed neither type 1 nor type 2 etching pat­
terns exclusively. These areas appeared as a generalized surface rough­
ening with regions resembling hollowed prism centres adjacent to areas
in which the pattern was more consistent with the loss of prism peri­
pheries. In addition, within these areas were regions in which the etching
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.376 Sieverstone/S axton/D ogon/F ejerskov

Fig. I. Scanning electron micrograph of an enamel surface that was exposed to

37-percent phosphoric acid for 1 min. The surface shows a pattern in which there
is a distinct hollowing of prism centres with relatively intact prism peripheries.
This type of damage, called type 1 etching pattern, was the most common seen in
this study.

Fig. 2. Scanning electron micrograph of an enamel surface also exposed to 37-

percent phosphoric acid for 1 min. However, in this specimen, the prism peripher­
ies have been removed preferentially, leaving the prism cores projecting towards
the original enamel surface. This type 2 etching pattern can be seen to extend
along the length of the rods, thus aiding in delineating individual prisms.
Fig. 3. Scanning electron micrograph of part of the surface seen in the pre­
vious figure, now at a higher magnification and viewed from the original surface.
Separate prism cores are seen, the gaps separating them corresponding to prism
peripheries. This is a type 2 etching pattern.
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378 Silverstone/S axton/D ogon /F fjerskov

Fig. 4. Scanning electron micrograph of an enamel surface exposed to 30-per­

cent phosphoric acid for I min. In this, areas can be seen showing a type 1 etching
pattern in which there is a preferential removal of prism cores. However, adjacent
regions show a type 2 etching pattern in which the reverse pattern can be seen. In
addition, in many areas the pattern of surface damage is difficult to relate to
prism structure. Such a field is referred to in this study as a type 3 etching pattern.

patterns did not apparently conform to prism morphology. These regions

were referred to as a type 3 etching pattern (fig. 4). Sometimes, the
whole region in the type 3 etching pattern was one in which the entire
surface topography could not be related to a prism pattern (fig. 5).

Fig. 5. Scanning electron micrograph of the inner fitting surface of a fissure

sealant (TP 226) [Silverstonf., 1974] after removal of the enamel by deminerali­
zation. This surface thus acts as a replica of the original etched enamel surface,
which had previously been exposed to 50-percent phosphoric acid for 2 min. In ad­
dition to some areas suggesting types 1 and 2 etching patterns, the topography of
the major part of the sealant cannot be related directly to prism structure. Thus,
this whole region represents a type 3 etching pattern.
Fig. 6. Scanning electron micrograph of an enamel surface exposed to 0.1 n
lactic acid for 48 h. The pattern of destruction is one in which there has been a
preferential loss of prism core material. Thus, this represents a type 1 etching pat­
tern and, due to the greater degree of tissue damage, is more clearly defined than
the similar pattern seen in figure 1. This was also found to be the most common
pattern of damage in the experiments with lactic acid.
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380 S ilverstone /S axton/D ogon/F ejerskov

Fig. 7. Scanning electron micrograph of an enamel surface also exposed to

0.1 N lactic acid for 48 h. However, a reverse pattern of damage is seen relative to
the previous figure, even though identical experimental conditions were employed.
Approximately 100 prisms can be seen in this field of view and they all show a
type 2 etching pattern in which the prism cores are left relatively intact, with a
preferential loss of prism peripheries.
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 Variation in the Pattern of Acid Etching 381

Fig. 8. Scanning electron micrograph of part of the enamel surface seen in the
previous figure, now at a higher magnification. In this type 2 etching pattern
prism cores are seen as bundles, clearly separated from each other by gaps repre­
senting the loss of prism peripheries. Individual arrays of crystallites can be identi­
fied in the prism core material, showing that the remaining tissue is relatively po­
rous.

The inner fitting surfaces of fissure sealants, when examined with the
scanning electron microscope, acted as accurate replicas of the acid-
etched enamel surfaces. All three types of etching pattern were seen in
these specimens. However, since the damaged surfaces were viewed in
replicas, a reverse pattern was observed (fig. 5). That is, with a type 1
etching pattern, the plastic replica appeared as a type 2 pattern. Similar­
ly the type 2 etching pattern, in which the prism cores were left stand­
ing, appeared as a type 1 pattern in the replica.
The rate of damage to the enamel surface, in terms of both depth of
etch and extent of porosity of the tissue, was found to be inversely pro­
portional to the concentration of phosphoric acid employed, as reported
previously [Silverstone. 1974].
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382 S ilvers TONE/SAXTON/DoGON/FhJtRSfcov

Fig. 9. Scanning electron micrograph of enamel which has been exposed to

0.001 n lactic acid for 96 h. The field of view shows prisms in a longitudinal
direction, demonstrating damage at the peripheral regions of the rods. Because of
this, the remains of the separate prisms can be seer quite clearly. Since the prism
core material is left relatively intact, with the loss of prism peripheries, this also
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 Variation in the Pattern of Acid Etching m

Lactic Acid
Exposure of enamel surfaces to lactic acid for periods of 1-5 days
produced a much greater degree of damage to the tissue than that found
with phosphoric acid. The enamel was etched to depths varying from
one quarter to three quarters of its entire thickness. With these experi­
ments, all three types of etching patterns were again seen, but this time,
the patterns were even more pronounced than in the previous series of
experiments.
Figure 6 shows a type 1 etching pattern in which there has been a
preferential loss of prism core material. The prism cores are much more
hollowed out than in the previous experiments using phosphoric acid,
thus demonstrating more extensive damage of the tissue. The average di­
ameter of the central depressions was 3-5 /<m. As with the experiments
employing phosphoric acid, this pattern was found to be the most com­
mon.
Figure 7 shows a specimen, also etched with lactic acid for 48 h, in
which the loss appears heaviest in relation to prism peripheries, and
therefore demonstrates a type 2 pattern of enamel etching. Approximate­
ly 100 prisms are seen in this field of view (fig. 7) and they all show an
identical pattern of attack. The prism cores are projecting 20-30 /tin
above the general level of the tissue clearly demonstrating a type 2 etch­
ing pattern of the enamel. The average width of the prism cores is
3 ,/<m. At a higher magnification (fig. 8) the crystallite bundles within
the prism cores can be seen clearly, indicating the extent of porosity of
the remaining tissue.
In figure 9, prisms can be seen in a more longitudinal direction. This
also demonstrates the type 2 etching pattern, the loss of prism peripher­
ies aiding in the identification of individual prisms. The damage to the
peripheral regions of the prisms is seen to extend for a significant dis­
tance along the prism structure.
Both type 1 and type 2 etching patterns were produced over the en­
tire concentration range of lactic acid used in these experiments, and
were apparently independent of exposure time.
Some regions were identified as showing a type 3 etching pattern, but
these regions were found to occur less often than when using phosphoric
acid. Figure 10 shows an area of inner enamel demonstrating a type 3

shows a type 2 etching pattern and is seen to extend a significant distance along
the prisms.
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384 S ieverstone /S axton/D ogon/F ejerskov

F/g. 10. Scanning electron micrograph of an enamel surface exposed to 0.1 n

lactic acid for 24 h. The surface shows a type 3 etching pattern in which there ap­
pears to be a random pattern of damage which is difficult to relate to prism struc­
ture.

etching pattern in which it is extremely difficult to relate surface topog­

raphy to prism structure. The various etching patterns produced were
not related to specific regions of enamel. That is, all three patterns were
found to occur on smooth enamel surfaces as well as on the occlusal
surface of the crown. In this, the results are in agreement with the find­
ings for phosphoric acid.

Discussion

When enamel was exposed to either phosphoric acid for short expo­
sure periods, or lactic acid with long exposures, patterns were seen in
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 Variation in the Pattern of Acid Etching 385

which there was a preferential loss of prism core material, called a

type 1 etching pattern in this study. This finding supports previous ob­
servations [M uller and Schait, 1957; P oole and J ohnson , 1967;
H offman et al., 1968, 1969a, b; J ohnson el al., 1971]. However, in
contrast to the above-reported findings, the reverse pattern of destruc­
tion was found in specimens exposed to identical conditions. That is,
prism core material appeared relatively intact with the preferential loss
of prism peripheries. This type 2 etching pattern was found in both se­
nes of experiments reported in this study. Although this second etching
pattern has been produced by the action of strong chelating agents on
enamel [P oole and J ohnson , 1967; H offman el al., 1968, 1969a, b] it
is obviously not unique to a chelator attack on human dental enamel as
has been inferred to date.
These results suggest therefore that there is no one specific etching
pattern produced by acid action on human dental enamel, as far as the
two acids used in this study are concerned.
P oole and J ohnson [1967] suggested that the different etching pat­
terns produced with either an acid, or a chelating agent, were due to dif­
ferences in orientation of crystallites relative to the direction of attack,
together with differences in chemical composition between central and
peripheral parts of enamel prisms. Little is known of differences in
chemical composition between these areas, and the fact that in this study
both etching patterns were produced by exposure to acid solutions, indi­
cates that differential solubilities between prism core and prism peri­
phery, if they exist, may be of little significance with respect to the pat­
tern of acid etching. Such differences, however, could be of significance
in the carious process which is a more diffusion-controlled attack rela­
tive to the relatively crude etching employed in these studies. In addi­
tion, the caries process consists of a dynamic scries of events, with phas­
es of both demineralization and remineralization [Silverstone, 1973],
rather than a more simple, continuing dissolution. Thus, since there is
an abrupt change in crystallite orientation at prism junctions, more in­
tercrystallite space exists. This space is probably filled with hydrated
organic matrix and so the organic content of the region will be higher,
even though no discrete organic prism sheath is thought to exist in
sound mature enamel [M eckel el al., 1965; J ohnson , 1967; Sundstrom
and Z elander. 1968],
The role of crystallite orientation may be significant in determining
differential etching effects in enamel. However, little is known regarding
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386 Silverstone /S axton/D ogon /F ejerskov

differences in the reactivity of different faces of individual enamel crys­

tallites to demineralizing agents. Previous studies [Johnson, 1966;
Sharpe, 1967; Boyde, 1971] suggest that enamel crystallites dissolve in
acid more quickly along their c-axes than in a direction perpendicular to
this axis. However, T yler [1969] has shown that calcium is liberated at
the same rate from areas on various faces of large fluorapatite crystals
exposed to acid.
J ohnson et al. [1971] suggested, as one further reason for the differ­
ences in etching patterns produced by either acids or chelators, that the
difference in size and electric charge exhibited by the different deminer­
alizing radicals might play a role. However, since in this study both
types of etching patterns were produced with acid solutions alone, this
tends to decrease the importance of this parameter as a likely explana­
tion for differences in action between the two types of agent.
This study has shown that the exposure of human dental enamel to
acid solutions in vitro produces three basic etching patterns. In the most
common, called type 1 etching pattern, prism core material was prefer­
entially removed leaving the prism peripheries relatively intact. In the
second, type 2 etching pattern, the reverse pattern was observed. The
peripheral regions of prisms were removed preferentially, leaving prism
cores remaining relatively unaffected. In the type 3 etching pattern,
there was a more random pattern, areas of which corresponded to
types 1 and 2 damage together with regions in which the pattern of etch­
ing could not be related to prism morphology. These findings differ from
previous studies in which the type 1 pattern was ascribed to acid action
and type 2 etching pattern to attack by chelators. The results therefore
suggest that there is no one specific etching pattern produced in human
dental enamel by the action of acid solutions. Such differences produced
by acids are difficult to explain on the basis of variation in chemical
composition and crystallite orientation. This further highlights the varia­
tion in structure that can occur in enamel not only from tooth to tooth,
or surface to surface, but also from site to site on a single tooth surface.

References

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Prof. L. M. S ilverstone, University of Minnesota, School of Dentistry. Minnea­

polis, MN 55455 (USA)
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