J. periodont. Res.

4: 83-93, 1969

The histology of the

muco-gingival junction
J. LOZDAN AND C. A . SQUIER

Department of Oral Medicine and Oral Pathology, The London Hospital Medical College,
London, U. K.

Nineteen pieces of muco-gingival tissue from man and monkey were examined by histological and
histochemical methods. Neither a surface groove nor a change from keralinisation to non-keratinisa-
tion were consistently seen in all specimens; the differing elastie tissue content of the gingiva and
alveolar mucosa was therefoie used to define the muco-gingival junction. The surface groove, the
keratinisation pattern, the distribution of hydrolytic enzymes and the glycogen content have been
related to this junction. The gingiva and alveolar mucosa meet wiEhout a gradual transition zone
suggesting that an intrinsic difference exists between them. This junction provides a useful model
system for the examination of epithelial-connective tissue interrelationships.

This study was undertaken in order to ex-
tend our knowledge of the biology of the
muco-gingival junction. This is of obvious
clinical importance to the periodontist who
frequently has to undertake surgical proce-
dures in this area.
There have been no reports in the avail-
able literature on the nature of the jtinc-
tion between gingiva and alveolar tnucosa
although a number of workers have exa-
mined the histology and histoehemistry of
these tissues in the same specimen (Wein-
mann et al. 1959, Weiss, Weinmann and
Meyer 1959, Turesky, Glickman and Fisher
1959, Kapur et al. 1963). Most of these
workers referred to the well established dif-
ferences between gingiva and alveolar mu-
cosa described by Orban (1948) without
actually defining the junction; a summary
of these differences is given iti Table L The
only precise definition of the muco-gingival
junction that has been found in the literature
is the clinical one given by Orban (1948)
who simply described it as a scalloped line
separating the gingiva from the alveolar
mucosa. No histological definition appears
to exist.
In view of this lack of adequate histo-
logical infomiation on the muco-gingival
junction, it was thought important to estab-
lish reliable criteria with which to define the
junction before examining further the bio-
logy of this region. Specimens of muco-
gingival tissue from both man and monkey
were examined by routine histological me-
thods. The distribution of two hydrolytic
enz3mies, acid phosphatase and nonspecific
esterase, which have been related to the •
process of keratinisation (Jarrett and Spear-
man 1964, Ten Cate 1963), was investigated
histochemically.
Materials and Methods
Tissue was obtained from both humans and
the Macaque irus species of monkey. Under

This paper was presented at the Meeting of the British Division of the I.A.D.R. in Cardiff, April
1968.
The authors wish to thank Dr. R. Duckworth for his help. Miss G. Davis assisted with the
preparation of the histological sections.
 LOZDAN AND SQUIER

MUCO GINGIVAL
'JUNCTION

CRYOSTAT CHUCK

Fig. 1. Diagram showing how tissue was removed and orferrtated. A, site ol biopsy; B, biopsy specimen; C,
specimen orientated on the cryostai chuck.

local anaesthesia twelve specimens were ob- at -25 °C and its temperature allowed to
tained from male and female patients with equilibrate. Sections were then cut at 8 /j
aa age range of 19-59 years of both Cau- and immediately fixed in formol-calcitim at
casoid and Negroid ethnic groups. The 4°C for 20 mios. After washing in saline
anaesthetic was injected at some distance the sections were ineubated in the follow-
from the site of the biopss'. A strip of ing media:
clinically normal muco-gingival tissue 1-2
mm vi'ide was taken at a site remote from 1) Sodium /J-glycerophosphate substrate for
the interdental papilla from patients under- acid phosphatase (Barka and Anderson
going periodontal surgery (see Fig. 1). The 1963) for 45-60 mins. at 37°C.
specimens were removed from the buccal
or labial aspects of teeth in all fotir quad-
rants of the mouth. Seven similar speci-
mens were obtained from monkeys utider Table I
general anaesthetic. Reported histological characteristics of the
muco-gingival tissues
All specimens were orientated on a cork
disc and supported by coagulated albumen attached giriQiva alveolar mucosa
(egg white) before being placed on a cryostat
keratinised non-keralinised
chuck (Fig. 1) and quenched in iso-pentane
cooled to -150°C in liquid nitrogen. thin, prominent flat, broad epithelial
epithelial ridges ridges
In order to prepare sections for histoche-
mical examination the chuck with mounted 00 elastic fibres in eiastic fibres in
specimen was placed in the cryostat cabinet corium — mucoperiosteum corium — mobile
 THE HISTOLOGY OF THE M U CO-G 1 N G i VAL JUNCTION 85

Table M
Summary of the results
Histological features within approximately 0.75 mm of the junction (i.e. within the area of a low power fieid)
are summarised. The symbols + and — indioafe only the presence or absence of a feature and do not re-
present quantitative vafues. G = gingiva; A.M. = aiveoiar mucosa; o = oTtho-keratinisation; p = parakeratlni-
sation; * = incomplete parakeratosis.

Epithelium Corium
V
Keratinis- PAS + ve Inflammat- Surface Eiaslic Inflammat- Accessory
Specimen
ataon: materiaf ory cells groove fibres ory cells structures

No. Source G A.f^. G A.M. G A.M. G A.ft-'l. G A.M. G A.M.

1 human +p —
2 human +P +p sahvary ducf
human
+
3 +P +p + -5-
+
4 monkey +0 — +• +
5 monkey +0 +0 -S- 4- muscle
6 monkey +0 — 4- —
7 human +p +p -f- 4"
8 human H-p +p 4- fat
9 humah +P +p
10 human +P .— + + -r nuscSe
11 human +P — + -1- -1-
12 human 4-p +p
13 human +P — + 4- -f
14 human +P — + 4- 4-
16 monkey +O — + + 4- musole
16 monkey +0 ~
17
18
monkey
monkey
+0
+0 —
.„ — -f -f

+
X
4-
+
4-
muscSe
muscle
19 human +p

2) Naphthol AS-MX acetate substrate for All sections were mounted in Canada
non-specific esteiase (Pearse 1960) for Balsam and examined with the light mi-
3-60 tnin. at room temperature (18"C). croscope.

Sections were then rinsed in saline and

mounted in glycerine jelly.
After cutting the cryostat sections the
remaining block of tissue was fixed in
formol-calcium, dehydrated, embedded in
paraffin wax and sectioned at 5 ^ . Consecu-
tive sections from each block were staioed
using the following techniques:
Haematoxylin and eosin, Van Gieson, Hart's
elastin stain (Schmorl 1921), Mallory's tri-
chrome stain, Masson-Fontana argentaffin
reaction. Periodic acid-Schiff (PAS), after
Me Manus (Pearse I960), incubation in
1% diastase solution prior to PAS reaction.
Results
The results described below are summarised
in Table II.
Hhtology
Although the tissue that was removed
was considered to be clinically normal, in-
flammatory ceils were invariably present in
the region of the muco-gingival junction
(see Table II). These cells were mainly
lymphocytes and could be seen in both the
corium and the epithelium (Fig. 2).
Clinically the muco-gingival junction ap-
pears as a scalloped line. In histoiogical
 LOZDAN AND SQUIER

AM

Fig. 2. A section throug'h the muco-ging:ival junction of monkey. The gingiva {G) is orlho-keratinised. the
alveolar m,ucosa (AM) it not keratinised; a deep surface groove is presonl. Some lymphocytes are visible
in the epithelium. Slamed H & E; x 1:50.
Nole: in ali' figrures the sections are orientated so that the gingiva (G) iies to the ieft and the alveolar mucosa
(AM) 10 the right. The site of the mucc-gingivai junction was determined by rele:ence to the elastic tissue
content ot the corium (as mentioned in the text) and is marked with a black bar.

Fig. 3. Low power view of the human muco-gingival Junction. The superficial layer ot the gingival epithelium
is parakeratinised and this extends past the junction into the alveolar mucosa (arrow). No surface groove
is present but there is a marked difference in the airangement of coiiagen in the corium of the gingiva and
the aiveolar mocosa. Stained H & E; x 60.
 THE HISTOLOGY OF THE M U CO-G ! NG I VAL JUNCTION 87

B ^-''^^^ffmM
Fig. 4 A. Lower power view of the muco-gingival junction of monkey stained with Hart's elastic stain and
nuciear fast red. The epitheiium is fully keratinised along its entire surface; elastic tissue is only preseni
in the corium of the alveolar mucoaa. x 75.

Fig. 4 B . Ah enlarged view of the area outiiined in Fig. 4 A. A sudiden abrupt increase in the number of eiastio
fibres present in the corium can be clearly seen; this coincides with the surface groove, x 135.
 LOZDAN AND S Q U I E R

Fig. 5. Human muco-gingiva! junction showing a siight overlap of keratinisation ot the type described as in-
complete parakeratosis. The most superiicial cells resemble the desp cetls of the epithelium and are not
keratinised, although some of the immiediately underlying cells stained orange and appear fuiiy
Stained wilh Mailory's tri-chrome; x 300.

sections this line may be represented by A. Surface groove pre.sent

a surface groove (Fig. 2) although in ap-
proxinnately half of the specimens examined
the groove was not identifiable (Figs. 3 and
6). As, Orban (1948) had already defined
the junction in terms of this groove, initially
we examined otily those specimens possess-
ing a dear histological surface groove. In
this way it could be determined whether
this point corresponded with a change in
the histological pattern from gingiva to
that of alveolar mucosa (see Table I). Later,
specimens without a surface groove were
examined.
Results are presented in two sections
depending on whether a surface groove was
present (A) or absent (B).
Epithelium
In the gingiva the epithelium was always
keratitiised (Fig. 2). The monkey specimens
were all ortho-keratinised. The human ging-
ival epithelium was para-keratinised (Fig.
3) although a definite distinction between
ortho- and para-keratinisation was often dif-
ficult to establish. In almost half of the
specimens, this keratinised layer extended
beyond the surface groove for a short
distance before true non-keratinised epithe-
lium was obvious (Fig. 3) and in some
monkey specimens the entire length of the
epithelium was ortho-keratinised* (Fig.4 A).
One human specimen showed an extension
of keratinisation from the gingiva to the

* These specimens were from the molar region and probably represent an extension of keratinisa-
tion from the cheek pouch into the alveolar mucosa. (Cohen; personal communication 1968).
 THE HISTOLOGY OF THE MUCO-GINGIVAL JUNCTION 89

AM

Fig. 6. The muco-gingival junction of a pigmented human patient after staining by the Masson-Fohtana ar-
gentaffin reaction; meianocytes extend beyond the junctioh inio the alveolar mucosa. A change in the
epitheilai ridgie pattern may aiso be seen., x 80.

alveolar mucosa side of the surface-groove
of the type described by Weinmann and
Meyer (1959) (Fig. 5) as incomplete para-
keratosis.
The long, thin epithelial ridge pattern
seen in the gingiva usually changed to a
broad flat pattern (see Fig. 6) at a point
coincident with the surface groove. This
change was not always seen nor easily
distinguishable in all sections.
Several specimens showed the presence
of melanin and dendritic melanocjrtes in
the epithelium after staining by the Masson-
Fontana argentaffin method. Althoug mel-
anocytes were mainly situated in the basal
layers of the gingival epithelium, some
could be seen in the alveolar mucosa at
some distance beyond the stirface groove
(Fig. 6). Melanin granules were present in
the epithelial cells adjacent to the melano-
cytes.
The distribution of glycogen was invest-
igated using the PAS technique with diastase
digestion of the control sections. In most
specimens little or no glycogen was present
in the gingival epithelium. Virtually all spe-
cimens, however, contained considerable
amounts of glycogen in the epithelium
beyond the surface-groove, as in Fig. 7.
Glycogen was never found to be present
in the gingiva when is was absent from the
alveolar mucosa.
Corium
There were obvious differences in the con-
nective tissue pattern on either side of the
surface-groove; the change in this pattern
was abrupt and always coincided with the
groove. CoOagen bundles were densely ar-
ranged in the gingiva btit were fewer and
more widely dispersed in the alveolar mu-
cosa (Fig. 3). In the gingiva virtually no
elastic fibres were present (except for those
associated with blood vessels) in contrast
with the large amounts seen throughout the
alveolar mucosa. This difference was strik-
m LOZDAN AND SQUIER

AM

Fig. 7, Human muco-gingival junction after treatment by the PAS technique. PAS positive material is located
primariliy in the epitheliuim o( the aSveolar mucosa; very little is present on Ihe gingival side of the junc-
tion. X 160.

Ffg. 8. Sectfon of muco-gingjva! junction from monkev after incubation for acid phosphatase according to
Barka and Anderson (1963). There is a marked change in the dis^:ributio.n of the enzyme at ithe junction
especialJy in the superficial layers, x 170.
 THE HISTOLOGY OF THE MUCO-GINGIVAL JUNCTION 91

AM

:<0:'--^^

Figi. 9 A. A section ihrO'Ugh the muco-gingival junctior of monkey. Note the abrupt termination of the kera-
tinised layer of the gingiva at the junction in this specimen. Stained H & E. x 150.

Flfl. D B. A section serial with that shown above (9 A) and incubated tO' stiow non-specific esterase activity.
The distribution of this enzyme in the epithelium changes abruptly at the junction. Fine elastic fibres
are obvious in the corium of the alveolar mucosa (airovi/s). Unstained; x 160.
 LOZDAN AND S Q U I E R
ingly demonstrated when elastic stains were
used (Fig. 4 A, B), but was also apparent in
unstained sections as the elastic fibres could
then be visualised by virttie of their re-
fractile properties (see Fig. 9 B, which is
an unstained histochemical preparation).
Accessory Structures
Various accessory structures were ob-
served and are noted in Table II. Bundles
of striated muscle were frequently seen in
the corium of alveolar mucosa from mon-
keys but only rarely in the human tissue.
This probably reflects a different location
of the orbiculoris oris muscle in the two
species.
B. Surface groove ab.ient
Although a surface groove was absent, a
change in the connective tissue pattern of
the corjum was always obvious within these
specimens as Fig. 3 demonstrates; the change
was similar to that seen when a groove was
present (see A, above). In a few specimens
the whole length of the epithelium was
keratinised, while in others there was a
change from keratinisation to non-keratinisa-
tion which did not always coincide with a
change in the underlying connective tissue
pattern.
Histoehemistry
A different distribution of both acid
phosphatase and non-specific esterase was
observed between the epithelium of the
gingiva and the alveolar niucosa; this dif-
ference has already been adequately de-
scribed (Ten Cate 1963, Kapur et al. 1963,
Cohen 1966). The change in the distribution
of each enzyme occurred at the point in the
epithelium marked by the surface groove (Fig.
8). A similar change was seen in those speci-
mens without a clear surface groove and in
these it occurred at a point in the epithelium
opposite the sudden change in elastic tissue
content of the corium. This is particularly
obvious in Fig. 9 where the histochemical
preparation for non-specific esterase (9 B)
can be compared with an histological sec-
tion through the muco-gingival junction
(9 A) of the same specimen.
Discussion
In order to define the muco-giogival junc-
tion it is necessary to distinguish between
gingiva and alveolar mucosa. Orban (1948)
defined the clinically recognisable muco-
gingival groove as the dividing line between
these tissues; in vivo this groove probably
arises from the differing attachment of the
gingiva and the aiveolar mucosa to under-
lying bone. A groove, however, was not
present in nearly half of the specimens,
probably because the tissue folded when
detached from bone. It is essential therefore
to seek more consistent criteria for an histo-
logical definition of the muco-gingival junc-
tion.
The results clearly show (Table II) that
neither the change in keratinisation, the
shape of the epithelial ridges,, nor the
change in composition of the corium were
coincident in most specimens. In some speci-
mens the only difference observed between
the gingiva and alveolar mucosa was the
differing structure of the corium.
Alveolar mucosa is a more mobile tissue
that the adjacent attached gingiva and there-
fore one would expect to find this func-
tional difference reflected structurally in the
corium of the two tissues. In fact a marked
difference in elastic tissue content was ob-
served in all specimens. It is therefore
reasonable to define the muco-gingiva] junc-
tion as that point where a sudden marked
increase of elastic fibres occurs in the co-
rium. The change in the distribution of acid
phosphatase, non-specific esterase and gly-
cogen took place at this junction. In a few
specimens (e. g. Fig. 9 A, B) the surface
groove, the change in the pattern of kerat-
 THE H I S T O L O G Y OF T H E M U C O - G I N G I V A L
inisation, and the change in the composi-
tion of the corium corresponded.
It is noteworthy that an abrupt change in
histological appearance and histochemical
reaction occurs at the muco-gingival junc-
tion, and this is particularly apparent in
the epithelium when enzyme histochemical
methods are used. This sudden transition
from one mucosa to another indicates the
individuality of each, and suggests the ab-
sence of any lateral influence in determining
keratinisation. Any controlling influence on
the epithelium of the muco-gingival tissues
must therefore arise from beneath, that is,
in the corium.
References
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mistry; theory, practice and bibliography.
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Cohen, L. 1966. Keratinisation in the oral mu-
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Jarrett, A. and R. L C. Spearman. 1964. Histo-
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Kapur, K.K., H. H. Chauneey, S.Shapiro and
G. Shklar. 1963. A eomparative study of the
enzyme histoehemistry of human edenEulous
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JUNCTION 93
Orban, B. 1948. Clinical and histologic study of
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Pearse, A. G. E. 1960. Histoehemistry; theore-
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Address:
Dept. Oral Medicine
London Hospital Medical College
Turner Street
London. E. 1.