COMPOSITION OF DENTAL ENAMEL /. A.
Weatherell
COMPOSITION OF DENTAL ENAMEL
J. A. WEATHERELL B.Sc. Ph.D.
Department of Oral Biology
Dental School, University ofLeeds
1 General chemistry of enamel
2 Minor constituents in enamel
3 Chemical variations within the enamel
4 Chemistry and caries
References
Interest in the chemistry of dental enamel is closely linked with
concern about the problems of dental caries. Recent work has
recognized the importance of enamel as an integrated biological
structure. Its heterogeneous nature cannot be understood
without appreciating what influence the process of enamel
formation has upon its subsequent chemistry and properties.
Also, if the pattern of carious attack is to be explained, infor-
mation must be available about any systematic chemical
variations within the enamel and their possible influence on
the rate of carious destruction. In this discussion, an attempt
has been made in the light of recent investigations (Weatherell,
Robinson & Hallsworth, 1974) to put the chemistry of enamel
in such a context.
1. General Chemistry of Enamel
The most mineralized of all biological tissues, with a density
much greater than the dentine upon which it sits (fig. 1),
enamel consists largely of inorganic material in the form of
small crystals. These are visible only in the electron microscope
and, for this reason, are usually called crystallites. The in-
organic material consists mainly of calcium phosphate and,
judging from x-ray and neutron diffraction data and from
infrared studies, it is essentially similar to the group of minerals
known to geologists as apatites (Kay, Young & Posner, 1964).
It is likely that calcium-deficient apatitic structures are present.
These might explain the sometimes low calcium: phosphorus
ratios, e.g. in the tooth surface (Wirsing, Judd & Ansell, 1974).
The possibility that minor amounts of other, non-apatitic
crystal forms exist in enamel has also been suggested: octa-
calcium phosphate (Brown, 1966), perhaps as a residue of
mineral present during the developmental stages of mineraliza-
tion, and dicalcium phosphate (Casciani, 1971) and (i-tri-
calcium phosphate (Vahl & Pladkova, 1967), perhaps as a
result of post-eruptive alterations brought about in the tooth
surface. It is thought, however, that any such non-apatitic
components of the mineral, if present, constitute a very small
part (under 2 % by weight) of the total inorganic material and,
from the point of view of its general chemical composition,
enamel can be considered to consist mainly of hydroxyapatite
(Young & Spooner, 1970).
The structure of the geological and synthetic apatites is
well established, and it has been presumed by analogy, together
with the results of chemical analysis, that the enamel mineral
115
Vol. 31 No. 2
consists chiefly of calcium, phosphate and hydroxyl ions in a
stoicheiometric ratio Ca10(PC>4)6(OH)2, each crystallographic
unit cell consisting of 18 ions arranged as hydroxyapatite.
A reactive part of the hydroxyapatite structure is the column
of hydroxyl ions parallel to the long axis of the crystal.
Invariably, some of these hydroxyl ions are missing, and this
allows increased ionic movement within the column, thereby
increasing its chemical reactivity. Other hydroxyl ions can be
replaced by fluoride; this probably restricts the mobility of the
hydroxyl ions in the column, resulting in stronger bonds with
adjacent calcium ions, making the crystal as a whole more
stable and facilitating crystal formation (Posner & Eanes,
1963). Calcium and phosphate ions can also be substituted.
2. Minor Constituents in Enamel
The fact that the enamel mineral is hydroxyapatite with this
capacity for ionic exchange ensures that the enamel contains at
least trace amounts of other elements, e.g. Na, Mg, Zn, K,
Pb, Sr, Fe, F and other minor ionic components, e.g. carbonate.
These can be incorporated while the apatite crystallites are
forming or by hetero-ionic exchange reactions at crystallite
surfaces.
Most of the minor ionic components in enamel are no doubt
properly considered as part of the apatite crystal structure,
but others probablyresideat least in part on the surfaces of the
apatite crystallites or as a separate phase. For instance, al-
though carbonate can be accommodated to a certain extent
within the lattice, replacing phosphate and to a lesser extent
hydroxyl ions, some of it may be situated at the crystallite
surface and could even exist separately as calcium carbonate
(Elliott, 1969). At least part of the magnesium and other ions
such as citrate, too large to fit easily into the lattice structure,
are probably outside the body of the crystallites.
FIG. I. Density of enamel and dentine, determined
by a floatation technique
3.0-1
1
S
ENAMEL| DENTINE (PULP
o
2.5-
a.
"5
2.0
Distance from
enamel surface to pulp (mm)
Eight specimens of enamel and 12 specimens of dentine were dissected
from » I OOnm-thlck section of a tooth
 COMPOSITION OF DENTAL ENAMEL /. A. Weatherell
The concentration of any particular ion in enamel seems to
depend directly upon its concentration in the environment
during development when the crystallites are forming and,
although generally to a lesser extent, upon its concentration in
the food and water ingested after tooth eruption. Its concentra-
tion also depends upon the rate at which the ion is absorbed
at the tooth surface, being influenced by the physical condition
of the enamel, its degree of mineralization and the pH at the
enamel-environment interface.
The best documented account of the factors influencing the
concentration of an extraneous ion in enamel concerns
fluoride and, since the factors governing fluoride uptake are
probably pertinent to the uptake and distribution of several
other ions, they can profitably be mentioned in some detail
here.
There is a well-established relation between the concentra-
tion of fluoride in the diet and drinking-water and its con-
centration in enamel (Brudevold, 1962). Any increase in the
availability of fluoride from extraneous sources, e.g. as a
result of the fluoridation of water supplies, the presence of a
silicate restoration, or the application of fluoride-containing
solutions to the tooth surface, is reflected by an increased
concentration of fluoride in enamel.
Fluoride is taken up most rapidly by enamel that is only
partially mineralized. Uptake is thereforerelativelyhigh during
the formative period, which is why, to be an effective public
health measure, fluoridated water supplies should be available
prior to tooth eruption. If, however, the enamel is incompletely
mineralized or becomes partially demineralized, even after
eruption, fluoride is again rapidly incorporated. This explains
the invariably high concentration of fluoride in carious lesions
or in other porous imperfections of enamel and would appear
to be the principal reason why a low pH at the tooth surface
enhances fluoride uptake; presumably the acid demineralizes
the enamel to some extent.
There are instances in which elements other than fluorine
have accumulated in enamel. Abnormal concentrations of lead
have been associated with cases of lead poisoning (Brudevold
& Steadman, 1956). Strontium is readily absorbed and the
well-known ability of teeth to accumulate 90Sr has been used as
a means of monitoring the extent of atomic fall-out (Starkey &
Fletcher, 1969).
Discussion of the mineral would not be complete without
some mention of the water present in enamel, which can
contribute up to 3 % of its weight. There is unfortunately no
clear consensus of opinion about the nature of this water,
despite numerous investigations, mainly with the use of nuclear-
magnetic-resonance techniques (e.g. Myers & Myrberg, 1965).
In one such investigation three types of water were reported,
some of which was free and some "freely tumbling water but
'caged' within the structure" (Little & Casciani, 1966). Other
workers have disagreed, however, with this description.
The older but still attractive concept of Neuman & Neuman
(1958) suggests that a small but chemically significant part of
the water might be present in the form of hydration shells
enveloping the crystallites. Representatives of many of the
ions mentioned above, particularly those associated with
outer regions of the crystallites, would be associated with the
water molecules in these hydration shells. Ionic exchange
reactions would occur most rapidly in this region.
It is also highly probable that some of the water in enamel
will be bound to the small amounts of proteinaceous material
present. Estimations of the actual amount of protein in the
116
mature tissue have arrived at widely differing values, from
0.36% (Eastoe, 1960) down to 0.06% (Glimcher, Friberg &
Levine, 1964; Weidmann & Eyre, 1967). In hindsight, the
most likely reason for the discrepancies is that most of the
analyses were carried out on samples pooled from large num-
bers of teeth. Sampling techniques varied and different regions
of enamel were studied.
3. Chemical Variations within the Enamel
Enamel is a heterogeneous structure, its composition varying
markedly from surface to interior and also from one region of
the tooth to another. In considering its nature and properties,
this cannot be ignored; it is essential to sample carefully defined
areas if chemical analyses of either the inorganic or the organic
components are to be meaningfully compared.
Analysis according to density, for instance, shows that the
total amount of material per unit volume always decreases from
surface to interior. The patterns of calcium and phosphate
distribution per unit volume of enamel are, as would be
expected, fairly similar to those of density, concentrations
decreasing from surface to interior. In general, over-all density
and mineral concentrations tend to be lower in the enamel
near the cervical region of the tooth, immediately beneath
the cusps, and below the fissures of molars and the pits of
incisors (Weatherell & Robinson, 1973). Mature enamel
consists almost entirely of apatite crystallites and presumably
the explanation for changes in density, calcium and phos-
phorus concentrations is that the amount of extra-crystallite
space varies, increasing towards the enamel-dentine junction
and in the other less highly mineralized regions.
The concentrations of minor inorganic ions also vary accord-
ing to consistent patterns, although not all constituents are
distributed in the same way. Concentrations of carbonate,
sodium and magnesium are relatively high in the interior and
diminish towards the surface. The reason for this is not clear
and there are several possible explanations.
If some of these ions are chiefly located at crystallite surfaces,
the lower concentration near the tooth surface might merely
reflect a decrease in the surface: mass ratios of the crystallites
in this region. The distribution pattern of sodium, on the other
hand (Besic, Knowles, Keller & Wiemann, 1970), could
indicate the distribution of water in the enamel.
Some of the distribution patterns might reflect events that
occurred during the period of enamel formation and early
mineralization. If, for instance, the carbonate of enamel were
to originate as carbon dioxide produced by the ameloblasts
as they advanced to the outer limit of the tissue during the
period of enamel formation, the amount of carbonate deposited
in the mineral would depend on the metabolic activity of the
enamel-producing cells. The more active they were, the more
carbonate would be incorporated into the forming mineral.
As their activity decreased towards the outer enamel, less
carbon dioxide would be produced and the amount trapped by
the forming enamel crystallites would therefore fall. A similar
explanation could account for the distribution pattern of
magnesium, if its origin, too, were related in some way to
cell function.
The distribution of most other trace elements is usually the
reverse of that associated with carbonate, sodium and magne-
sium, concentrations being highest near the tooth's surface
and decreasing towards the interior. These ions are mainly
exogenous in origin and accumulate principally by ionic
Br. Med. Bull. 1975
 COMPOSITION OF DENTAL ENAMEL /. A. Weatherell

FIG. 2. Fluoride distribution in the enamel of FlG. 3. Contours of density (expressed in g/cm1) in
lOOum-thick section from a human molar an enamel section from the cusp of a permanent
molar

The fluoride content of small enamel particles dissected from the

section was determined by use of the Orion fluoride electrode

exchange at the tooth's surface. As mentioned in section 2, from
the manner of fluoride uptake it appears that most of this
exchange takes place before eruption, when enamel is relatively
permeable. The rate of incorporation slows down as enamel
becomes more mineralized. Once enamel is fully mature,
probably within, say, two years after eruption, the rate of
uptake is considerably reduced. The chemistry of the bulk of
the enamel is therefore mainly established before eruption and
seems to remain essentially unaltered throughout the tooth's
life. Only in unusual circumstances, if for instance concentrated
solutions of ions are applied to the tooth, is the composition
of the sound mature enamel drastically altered.
In the enamel interior, the concentration of most of these
trace elements is relatively low although, again, apart from
fluoride studies there is little detailed information about any
internal variations. In the case of fluoride, where enamel den-
sity is low, e.g. near the amelo-dentinal junction, concentra-
tions of the element tend to be raised (fig. 2). The probable
reason is that, where enamel density is low, there is more inter-
crystallite spacefilledwith organic material and water, and the
enamel in such a region is more permeable. The same con-
protein of oral epithelium or stratum corneum than to the
proline-rich organic matrix of forming enamel. It seems pos-
sible that the tuft material in mature enamel originates during
development, but perhaps constitutes part of the protein laid
down before the ectodermal cells of the inner enamel epithelium
have fully differentiated into amelogenin-secreting ameloblasts
(Robinson, Lowe & Weatherell, 1975).
There is relatively little organic material in the main body
of the enamel but the concentration often rises again at the
enamel surface. This is, however, attributable to the presence
of an organic pellicle consisting of oral debris and, in those sites
protected from wear, perhaps including residual cellular mater-
ial from the layer of ectodermal cells which originally formed
the inner enamel epithelium. This organic layer, whether on
FlG. 4. Fluoride concentration (expressed in p.p.m.)
in different areas of a premolar from a 44-year-
old subject
Severe wear
200
2000T

siderations will probably apply to internal variations in the

concentration of other trace elements.
On a dry-weight basis, calcium and phosphorus gradients 1500
are more variable than on a volume basis and this itself is
perhaps an indication of the variations in the amount of organic
material situated in the space between the crystallites. The 1000-
amount of organic material present in mature enamel is some- BUCCAL
what small for accurate detailed analysis to be carried out
although qualitative information about its distribution has 500
been obtained from histological studies, and there have been
some attempts to determine its distribution quantitatively
(Wachtel, 1959; Robinson, Weatherell & Hallsworth, 1971).
Relatively large amounts of protein are present near the dentine,
apparently associated with the structures known to histologists
as enamel tufts; this is no doubt why enamel density sometimes
falls steeply near dentinal cornua (fig. 3). In vivo, there is prob-
ably also water bound to protein in this area, although the
details of the mineral-water-organic material relations in
enamel are not known and, at such low concentrations, the
direct determination of water introduces considerable technical
problems.
The relatively large amounts of protein present near the
enamel-dentine junction are probably a residue of the organic Loss of penkymata
matrix of forming enamel, most of which is withdrawn during Each curve represents the gradient In fluoride concentration from
the process of mineralization. Amino acid analysis, however, surface to interior of enamel, each point on the curves representing
reveals a composition somewhat closer to the ectodermal the fluoride concentration in a layer of enamel about 20jim thick

117

Vol. 31 No. 2
 COMPOSITION OF DENTAL ENAMEL /. A. Weatherell
the tooth surface or in fact a part of it, might protect the
enamel from carious attack and is in this sense functionally
part of the tooth, although it should not strictly be regarded as
part of the true enamel structure.
The original surface region of the enamel is, however,
clearly different from the interior in a number of other ways.
The tendency for extraneous elements to accumulate by ionic
exchange makes the composition of the outer region, say
30 (xm, significantly different from that of the deeper regions.
The gradients in density, calcium, phosphate, carbonate and
magnesium concentrations also give the surface special chemi-
cal characteristics.
The composition of the surface is not static; it changes
constantly with age. In some areas, particularly where the
enamel surface has been affected by the low pH of an overlying
plaque, fluoride and probably other trace elements increase in •
concentration. Elsewhere, slight abrasion gradually removes
much of the original surface together with a large proportion
of the tooth's high-fluoride enamel. In districts where the
fluoride content of the drinking-water is low, the high fluoride
concentration in the enamel surface can be irrevocably lost,
although in high-fluoride districts it might to some extent be
restored by post-eruptive uptake of fluoride from the saliva.
Where abrasion is severe, most of the enamel is worn away and
the new surface has the low density, high carbonate, high
magnesium and low fluoride concentrations characteristic of
interior enamel (fig. 4).
4. Chemistry and Caries
Investigations into the chemical composition of dental
enamel have not been carried out merely as an academic exer-
cise. Attempts to control dental caries require information
about any relation between the quality of enamel and its
resistance to carious attack.
It is because of the special clinical dental interest in the
benefits of fluoridation1 that the uptake and distribution of
fluoride in enamel has received so much attention. The most
widely accepted explanation for its anti-caries effect is that it
makes the enamel mineral more resistant to acid produced by
bacteria at the tooth surface. As mentioned in section 1,
fluoride can exchange with hydroxyl ions in the crystal lattice.
This stabilizes the structure and facilitates apatite formation,
probably by increasing the degree of hydrogen bonding in the
hydroxyl column. It has been shown experimentally that
fluoridated apatite is more crystalline, at any rate in bone
(Posner & Eanes, 1963), and it seems likely that fluoridated
crystallites tend to be larger, with a smaller surface area per
mass of mineral. All this reduces the ease with which enamel
mineral dissolves in the acid produced by bacterial fermenta-
tion.
The highfluorideconcentration in the outer region of enamel
has, not surprisingly, been regarded as a principal defence
against carious attack. There is an established positive correla-
tion between the fluoride content of enamel and the incidence
of dental caries, although the work of Brudevold (1962)
suggests that a concentration of about lOOOp.p.m. in the outer
region of the enamel gives optimal protection against caries;
and it is worth noting that higher concentrations can be asso-
ciated with abnormal, fluorotic, enamel.
» See Jenkins, pp. 142-145 of thli Bulletin.—ED.
118
It has been mentioned in section 3 that much of the fluoride
originally present in the outer region of enamel of a newly
erupted tooth is subsequently removed by wear. This is no
great disadvantage so far as caries is concerned, however, for
such loss of fluoride does not occur in those sites most suscep-
tible to carious attack; the most susceptible sites are the so-
called "stagnant" regions of the dentition where abrasion is
minimal. In such sites, fluoride concentrations probably
increase with time, particularly when the fluoride content of
the diet and the drinking-water isrelativelyhigh. Such stagnant
regions are also usually covered by plaque, an organic layer
produced by and including oral bacteria trapped within it2.
The acid produced by these plaque bacteria lowers the pH at
the tooth surface. This can destroy the enamel but it also
increases the rate of fluoride incorporation, partly because the
acid increases the enamel's porosity and therefore the surface
area available for exchange reactions, but probably also because
the fall in pH itself increases the rate of fluoride exchange
per se. This mechanism may be extremely important, for an
acceleration of fluoride uptake at the site of attack might halt
or even reverse the process of enamel destruction. Indeed, it
seems possible that the chief value of topical fluoride treatment
is to introduce fluoride to the forming lesion, the amount
absorbed being much larger and probably more significant so
far as caries inhibition is concerned than that taken up by
sound enamel. Recent work by J. L. Hardwick might be
similarly explained. Hardwick (personal communication)
found in experiments with rats that caries inhibition was
greater when the fluoride was added to the diet than when it
was administered in the drinking-water, despite the fact that,
in general, fluoride is absorbed more easily from drinking-
water. A possible explanation is that the fall in pH induced
by the large quantity of sugar in the cariogenic diet would cause
the plaque pH to fall and accelerate the uptake of fluoride into
underlying incipient lesions. It is also of some interest to note
in this connexion that cervical regions of teeth are not especially
prone to caries, despite the fact that these regions tend to be
covered by plaque and the enamel usually shows signs of
demineralization or change. In fact, this region of enamel tends
to have a high and apparently increasingfluorideconcentration,
no doubt partly because of the effect of the low plaque pH;
and this high fluoride concentration may be one reason why
cervical enamel is relatively resistant to caries.
The carbonate ion, which tends to introduce disorder into
the apatite crystal lattice, has also received some attention.
Carbonate is smaller than phosphate and when it replaces
phosphate will tend to reduce lattice dimensions. It is, on the
other hand, bigger than hydroxyl and when itreplaceshydroxyl,
the lattice will expand (Elliott, 1969). The presence of carbonate
in enamel, probably for this reason, introduces strain into the
structure of the apatite lattice and increases the ease with
which the enamel mineral dissolves in acid.
Carbonate was regarded by both Hardwick (1949) and by
Sobel (1962) as a possible Achilles' heel, enabling acid to
penetrate the enamel structure. There is no definitive proof
that this is the case, although acid and carious attack both seem
to remove carbonate preferentially (Johansen, 1962). Recent
analyses of samples of enamel dissected from thin sections of
carious lesions (Hallsworth, Weatherell & Robinson, 1973)
suggested that the earliest histological changes were associated
* See Hmrito & Bowden, pp. 131-136 of thh Bulletin.—ED.
Br. Med. Bull. 1975
 COMPOSITION OF DENTAL ENAMEL J. A. Weatherell
with the removal of mineral rich in both carbonate and mag-
nesium. Perhaps Hardwick's and Sobel's Achilles' heel should
have included magnesium as well as carbonate. It is unfortu-
nately not known where carbonate and magnesium are located
in the enamel structure, but the amount dissolved during the
initial attack seemed too great to be explained by a direct
stoicheiometric dissolution of hydroxyapatite. If an initial
inroad into the mineral fraction were to be made by the
removal of a small amount of non-apatitic mineral, access to
and dissolution of the remaining apatitic material could be
greatly facilitated.
At a histological level, caries appears to penetrate enamel
via the striae of Retzius, passes along the interprismatic
pathways into the cross-striations of the prisms and, finally,
into the prism cores (Darling, 1959). It has been suggested
from qualitative histological studies that these pathways
contain relatively high concentrations of organic material. It
also seems possible that the carbonate- and magnesium-rich
mineral lost from the translucent zone is removed from these
pathways, judging from circumstantial histological evidence
obtained by examination of areas sampled by Hallsworth
et al. (1973). Definite evidence is, however, lacking and it may
well be that the first mineral to be removed from the tooth during
carious attack is not associated with any visible histological
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VoL 31 No. 2
site. Johnson (1966) and J. Arends (personal communi-
cation), for instance, have shown that the process of carious
attack and the acid destruction of enamel crystallites in vitro
involve the preferential dissolution of crystallite cores. Arends
suggests this is explained by the presence of linear dislocations
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enamel crystallites, although, as he points out, the possibility
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from the remainder cannot at present be ruled out. Here, as
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it is often extremely difficult to distinguish between the influence
of chemical composition and the possible effect of structural
variations on the progress of attack. As indicated in this paper,
however, chemical composition and tooth surface structure
are inextricably linked and it is essential to consider the
chemistry of enamel within its developmental, histological
and anatomical context.
ACKNOWLEDGEMENTS
I should like to acknowledge the fact that the original work
described in this paper was carried out jointly with other members of
the Department of Oral Biology at the Dental School, University of
Leeds. I also wish to thank the Medical Research Council of Great
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