Mast Cell Distribution in Oral Tissues of Germ-free vs. Conventional Beagle Dogs
received suitable v i t a m i n and m i n e r a l supplements and continued sterility o f the germ-free a n i m a l s was deter m i n e d by weekly cultures o f fecal swabs and s e r o l o g i c a l assays o f serum from c o m m o n canine v i r a l antigens. A l l animals were sacrificed between the ages o f 10 to 12 months, with the exception o f one c o n v e n t i o n a l a n i m a l sacrificed at the age of 14 months. A l l germ-free a n i m a l s remained sterile u n t i l time o f death.
by
Immediately after
sacrifice,
the j a w s and
selected
tissue biopsies were fixed i n phosphate buffered f o r m a l i n
CHARLES M . COBB*
( p H 7.4) for a p p r o x i m a t e l y 12 days. A f t e r fixation, b l o c k
JAMES B . H E N E G H A N I
sections o f the right and left second m a n d i b u l a r bicus
DENIS M . L E B L A N C ‡
pids were processed for study by light m i c r o s c o p y . O t h e r tissues chosen for study included sections from
M I C H A E L J . DAVIS‡
the b u c c a l m u c o s a , cervical l y m p h node, s u b m a n d i b u l a r and p a r o t i d glands, and the m i d d l e one-third o f the
EHRLICH
1
tongue. A l l tissue was embedded i n paraffin and sec
is generally credited w i t h being the first to
tioned at 6 µ. A l t e r n a t e sections were stained respec
describe the mast cell. H o w e v e r , the granulated cells, known
as
mast
Recklinghausen
2
cells,
were
actually
described
tively with hematoxylin-eosin and with
by
1%
aqueous
toluidine blue.
3
and W a l d e y e r before E h r l i c h defined
them by their m e t a c h r o m a t i c granules. T h e term " m a s t
Biopsies of buccal m u c o s a and m a r g i n a l gingivae from
c e l l " originated from the overladen granules, which give
one germ-free and one conventional d o g were processed
the cells the appearance o f being overfed. In 1894, U n n a
for routine e x a m i n a t i o n by electron m i c r o s c o p y . C o l d
4
phosphate buffered gluteraldehyde was the i n i t i a l fixative
described a mastocytosis o f s k i n in u r t i c a r i a pigmentosa; since then significant i n f l a m m a t o r y , h e m o d y n a m i c , and
and cold 2% o s m i u m tetraoxide was used for postfixa-
hemostatic activities o f mast cells have been extensively
tion. T h e tissue was dehydrated in a graded series o f
explored. Despite a history that spans a century o f study,
alcohols and embedded in E p o n b l o c k s . T h i n sections
the b i o l o g i c functions o f the mast cell r e m a i n relatively
were
obscure.
acetate.
double
stained
with
lead
citrate
and
uranyl
5,6
It is widely accepted that the host response to bacterial
M a s t cells were counted at a m a g n i f i c a t i o n o f x 250
plaque a n d / o r plaque products is i n f l a m m a t o r y and that
using a grid that bounded an area o f 0.3 m m x 0.3 m m
this response is the major destructive factor i n perio
(0.9 m m ) . S i x sections were used from each tissue biopsy
d o n t a l disease. T h e m a n y c h e m i c a l mediators o f the
from each a n i m a l , and 12 fields were counted in each
2
i n f l a m m a t o r y process associated w i t h mast cell p o p u l a
section. C o u n t i n g of overlapping fields was avoided and
tions i n o r a l tissues have led to numerous investigations
only those cells that were u n m i s t a k a b l y identifiable as mast cells were counted. C e l l counts were converted to
and an extensive literature.
the number o f cells per square m i l l i m e t e r o f tissue.
T h e present report was m o t i v a t e d by the lack o f infor
A second evaluation was made o f the m a r g i n a l g i n
m a t i o n c o n c e r n i n g mast cell populations i n various o r a l tissues o f germfree animals, p a r t i c u l a r l y i n the beagle
givae associated with the
d o g , w h i c h is susceptible to p e r i o d o n t a l disease in the
bicuspids. T h e gingivae were graded as to the degree o f
m a n d i b u l a r right and
left
c o n v e n t i o n a l e n v i r o n m e n t and has become a m u c h used
i n f l a m m a t o r y infiltration using a scale o f slight, moder ate and severe. M a s t cell counts were taken from the area
e x p e r i m e n t a l m o d e l in p e r i o d o n t a l research.
just below the epithelium. A s i n the i n i t i a l cell counts, 12 MATERIALS AND METHODS
fields were counted in six tissue sections from each tooth from each a n i m a l . T h e collected data were evaluated for
S i x germ-free beagles (obtained by C a e s a r i a n deliv ery) and four c o n v e n t i o n a l a n i m a l s were acquired from the L o u i s i a n a State U n i v e r s i t y M e d i c a l C e n t e r , N e w O r l e a n s , L o u i s i a n a . T h e germ-free a n i m a l s were raised on a diet o f sterilized (autoclaved) B o r d o n ' s E s b i l a c P u r i n a D o g C h o w and water ad libitum. T h e conven t i o n a l a n i m a l s , w h i c h served as controls for this study, were fed the same diet i n nonsterile f o r m . A l l animals
differences between means by using the Student's t Test. A n a l y s i s for statistical significance c o m p a r e d the means within i n f l a m m a t o r y groups, i.e., slight vs. moderate, slight vs. severe, and moderate vs. severe. RESULTS
E x a m i n a t i o n o f mast cells in a l l tissues by light microscopy revealed the presence o f four basic cell morphologies: r o u n d , o v o i d , spindle or elongated, and pseudopodial. A l l four cell forms have been described in the l i t e r a t u r e . A s previously noted by R o b i n s o n and D e M a r c o , the density o f the connective tissue appeared to
* Department of Periodontics, University of Alabama-Birmingham, University Station, Birmingham, Alabama, 35294. † Louisiana State University, Medical Center, New Orleans, La 70112. ‡ 3320 North Hullen Street, Metairie, La 70002.
7-9
9
230
Volume 47 Number 4
Mast Cell Distribution
determine w h i c h o f the four basic cell forms was p r e d o m i n a n t in any given tissue. T h e elongated cell forms were more prevalent in dense connective tissue whereas rounded cell forms were preponderant in loose connective tissue areas ( F i g . 1 and inset). U l t r a s t r u c t u r a l e x a m i n a t i o n revealed mast cells to have the characteristic, numerous projections from the cell m e m b r a n e , pseudopodia, and m i c r o v i l l i , which are seldom seen i n other granulated cell t y p e s . E a c h cell observed contained a single nucleus located in an eccen tric position. T h e usually irregular nuclear m a r g i n was a result o f adjacent c y t o p l a s m i c granules ( F i g . 3). F e w 10
231
G o l g i structures were noted and the e n d o p l a s m i c reticu l u m were also sparse. T h e c y t o p l a s m contained numer ous fibrillar structures ( F i g . 2), diffusely distributed ribosomes and scattered glycogen granules. M a s t cells contained specific granules that varied from those o f homogeneous density ( F i g . 3) to those with coils and l a m i n a t e d structures ( F i g . 2). T h e latter structures rep resented a well-organized a r r a y o f p a r a l l e l , alternate dense and less dense lines with a periodicity o f a p p r o x i mately 75 to 100 A. T a b l e 1 presents the quantitative d i s t r i b u t i o n o f mast cells in the various o r a l tissues o f both germ-free and
FIGURE 1. Sections (1 n thick) stained with toluidine blue demonstrating mast cell population in loose connective tissue of the buccal mucosa from a germ-free beagle dog. Note perivascular location (V) and fact that most cells represent the round and/or oval morphology classification (insert). (Original magnification 250 x , insert 800 x . )
FIGURE 2. A section through the cytoplasm of a mast cell from the buccal mucosa of a germ-free beagle dog. The cytoplasm is densely packed with a heterogenous population of granules exhibiting coiled and laminated internal structure. Note the numerous cytoplasmic vacuoles (V) and fine filaments (arrows). (Original magnification 30,000 x . )
232
J. Periodontol. April, 1976
Cobb, Heneghan, LeBlanc, Davis
F I G U R E 3. A section through the cytoplasm of a mast cell from the buccal mucosa of a conventional beagle dog. The cytoplasm exhibits many membrane bound granules (arrows), each with an amorphous matrix of uniform density. Several cytoplasmic vacuoles were observed and occasionally a lipid droplet (L). Note also how the nuclear membrane ( N M ) presents an irregular contour due to proximity of the granules. (Original magnification 33,000 x.)
2
TABLE 1. Mast Cell Counts (cells/mm ) and Standard Deviations (S.D.) in Various Oral Tissues of Germfree vs. Conventional Beagle Dogs Germfree Cells/mm
2
Conventional
S.D.
Cells/mm
2
S.D.
Tongue Buccal Mucosa
669.3 449.0
±126.5 ±110.2
598.9 487.0
±111.2 ±128.2
Marginal Gingiva
305.5 88.4
±80.8 ±33.3
324.0 101.7
±91.0 ±37.6
106.0
±35.9
73.7
±19.9
±9.9
51.7
±12.8
Parotid Gland Submandibular Gland Cervical Lymph
38.1
Node * All values represent means ± S.D.
c o n v e n t i o n a l a n i m a l s . It should be noted that the counts obtained for s i m i l a r tissues from both types o f a n i m a l s were not significantly different. T h e mast cell p o p u l a t i o n o f the tongue was evenly distributed between the connective tissue adjacent to surface e p i t h e l i u m a n d that associated with the tongue musculature. M a s t cells o f the salivary glands were confined to the connective tissue s t r o m a and were usually adjacent to c i r c u l a t o r y vessels. In b u c c a l m u c o s a a n d m a r g i n a l gingivae the mast cell populations were d i s t r i b uted throughout the l a m i n a p r o p r i a with no p a r t i c u l a r pattern o f c o n c e n t r a t i o n . In contrast, cervical l y m p h nodes c o n t a i n e d relatively few mast cells a n d most were concentrated at the periphery o f the node and adjacent to the connective tissue capsule. E x a m i n a t i o n o f 20 teeth with their m a r g i n a l gingivae (from six germ-free a n d four conventional a n i m a l s )
F I G U R E 4. Example of degranulating mast cell from severely inflamed marginal gingiva of a germ-free beagle dog. A I µ-thick section from Epon embedded tissue stained with toluidine blue. (Original magnification 800 x.)
showed the d i s t r i b u t i o n o f the three i n f l a m m a t o r y groups to be o f r a n d o m pattern. T h u s , no c o n c l u s i o n c o u l d be made with respect to degree o f i n f l a m m a t i o n o f m a r g i n a l gingivae i n germ-free vs. the c o n v e n t i o n a l beagle d o g . N o a n i m a l was completely free o f i n f l a m m a t i o n at the m i c r o s c o p i c level, c o i n c i d i n g with a previous observation made by L i s t g a r t e n a n d H e n e g h a n , w h o used a n i m a l s from this same germ-free c o l o n y . T h e m a r g i n a l gingivae o f three teeth exhibited severe i n f l a m m a t o r y infiltration as well as p r o n o u n c e d food a n d / o r hair i m p a c t i o n . A l t h o u g h not a quantitative observation, it was noted that i n the severe i n f l a m m a t i o n group, m a n y of the mast cells appeared to be degranulating ( F i g . 4). T h i s was not true o f mast cells associated w i t h the slight and moderate i n f l a m m a t o r y groups. T a b l e 2 presents the 1 1 , 1 2
Volume 47 Number 4
Mast Cell Distribution
233
2
TABLE 2. Mast Cell Counts of Marginal Gingivae According to Inflammatory Status {cells/mm ) Inflammatory Groups Slight
Moderate 11
6
Number of specimens in
Severe 3
group 3154
1993
Total number of mast cells
388
counted Mean count Standard deviation Range of count Statistical significance
332.17
286.73
129.33
+58.95
+46.85
+92.52
206-421
148-406
Slight vs. moderate
Moderate vs. severe
Severe vs. slight
None
None
None
results o f data analysis concerning the means o f mast cell counts o f m a r g i n a l gingivae a c c o r d i n g to i n f l a m m a t o r y status. N o t e that the difference between the means and, therefore, the difference i n n u m b e r o f mast cells is not significant when any t w o groups are considered. T h e o v e r a l l tendency was for a decrease i n the number o f mast cells as i n f l a m m a t i o n increased i n severity. DISCUSSION
M a s t cells, w h i c h are inherent i n the connective tissue o f m a n y organ systems, routinely exhibit a perivascular l o c a t i o n , an association thought to favor release o f their g r a n u l a r contents into the c i r c u l a t o r y system. B a s i c aniline dyes, such as methylene blue, toluidine blue o r cresyl violet are classically used to demonstrate the presence o f mast cells. These dyes elicit a m e t a c h r o m a s i a of specific c y t o p l a s m i c granules, a characteristic that stems from the strong negative charge o f the sulfated carbohydrate components o f the granule contents. T h e positive staining o f granules by the P A S reaction, alcian blue, astra b l u e , or colloidal i r o n suggests the presence o f acid m u c o p o l y s a c c h a r i d e s . These h i s t o c h e m i c a l considerations can be explained by the large a m o u n t of heparin i n the g r a n u l e s . T h e difference i n staining reactions o f mast cells w i t h i n the same tissue a n d / o r between tissues suggests either a heterogeneity o f c h e m i c a l constituents o r a difference i n cell m a t u r a t i o n . 13
14
1 5
16
17
In this study, the light a n d electron m i c r o s c o p i c observations o f mast cells indicate that the general m o r p h o l o g y o f this cell type, i n germ-free a n d conven t i o n a l beagle dogs, resembles that found i n m a n by other investigators. T h e four basic cell shapes described in the l i t e r a t u r e , as well as s i m i l a r u l t r a s t r u c t u r a l characteristics, were observed i n tissues o f both g e r m free a n d c o n v e n t i o n a l a n i m a l s . T w o basic granule m o r phologies were observed: one type presented an a m o r phous m a t r i x o f u n i f o r m density, a second type exhibited l a m i n a t e d coils o f various sizes a n d densities. T h e t w o types o f granules were present i n both elongated and o v a l shaped cells. S i m i l a r observations have been reported i n studies o f the specific granules i n mast cells o f other animal species. 9 , 1 8 , 1 9
7 , 8
20,22
In this study, the d i s t r i b u t i o n o f mast cells i n various
99-156
tissues (Table 1) agrees with the investigations o f M o t a , et a l . , A r v e y , et al., and T o d a r o , who used dogs as the e x p e r i m e n t a l a n i m a l . In most a n i m a l species, the tongue and b u c c a l m u c o s a appear to be relatively rich i n mast cells, whereas salivary glands a n d l y m p h nodes exhibit scant p o p u l a t i o n s . In this respect, the g e r m free beagles o f this study showed no v a r i a t i o n from what is currently accepted as the n o r m a l pattern o f d i s t r i b u tion. 23
24
2 5
2 3 l 2 6
Because mast cells are a n o r m a l cellular component o f healthy gingivae, both a n i m a l a n d h u m a n , nu merous investigations have sought to correlate the n u m ber o f mast cells with the degree o f i n f l a m m a t o r y involvement. T h e conflicting results produced by these studies is due, i n part, to differences i n histologic technique and a lack o f adequate controls. T h e i m p r o v e d techniques a n d m o r e precise controls used i n recent investigations have yielded m o r e consistent results. These results demonstrate that the number o f mast cells decreases d u r i n g acute i n f l a m m a t i o n , possibly because degranulation renders them undetectable by the usual histologic s t a i n s . I n contrast, the mast cell p o p u l a t i o n in areas o f c h r o n i c i n f l a m m a t i o n exceeds that i n healthy g i n g i v a e . In this study, the tendency for a decrease o f mast cells that accompanies increased i n f l a m m a t i o n m a y be explained b y a c o m b i n a t i o n o f t w o factors: (a) m a r g i n a l gingivae o f three teeth exhibiting the most severe i n f l a m m a t o r y infiltration were also characterized by the acute phase o f the i n f l a m m a t o r y reaction; (b) only cells definitely identified as mast cells were counted, thus faintly stained cells were not included i n the t o t a l count. In his study, Z a c h r i s s o n counted lightly stained cells, considering them as i m m a t u r e mast cells, which explains his finding o f an increase i n mast cell p o p u l a t i o n d u r i n g c h r o n i c g i n g i v a l i n f l a m m a t i o n . A s regards acute i n f l a m m a t i o n , A n g e l o p o u l o s has stated that few mast cells are found i n areas infiltrated by p o l y m o r p h o n u c l e a r leukocytes. 2 3 l 2 5 , 2 7 - 2 9
27,30
31
31
3 1
3 2
Besides being i m p l i c a t e d i n gingivitis, mast cells m a y contribute t o development o f lesions o f other o r a l m u c o u s membranes. U p o n finding that the mast cell p o p u l a t i o n was significantly reduced 48 hours after the c l i n i c a l onset o f M i k u l i c z ' s recurrent o r a l aphthae,
234
J. Periodontol. April, 1976
Cobb, Heneghan, LeBlanc, Davis
Dolby and A l l i s o n
3 3
suggested that the reduction m a y
represent a T y p e I V allergic reaction. H a l l
3 4
reported a
seemingly specific pattern o f mast cell d i s t r i b u t i o n i n the following
mucous
membrane
diseases:
lichen planus,
p e m p h i g o i d , a n d desquamative gingivitis. J a n d i n s k i , et 35
al.,
suggested that the increased number o f mast cells
present i n low-grade c a r c i n o m a m a y result from an i m m u n e response t o antigenic s t i m u l a t i o n . A s yet, the precise role o f the mast cell i n i n f l a m m a t i o n has not been elucidated. H o w e v e r , several good reviews o f the possible b i o l o g i c functions have published.
3 6 , 3 7
been
A recent finding, the synergistic role o f
the mast cell a n d an increased activity o f m a m m a l i a n collagenase, deserves further investigation a n d m a y be found to account for the l o c a l i z e d collagenolytic activity in p e r i o d o n t a l pocket f o r m a t i o n . S i m p s o n and T a y l o r
3 8
reported that the in vivo c o n t r o l o f collagenase is, i n part, a factor o f e n z y m a t i c function a n d that the enzymes i n v o l v e d are elaborated b y mast cells. These mast cell factors/enzymes
reversed the i n h i b i t i o n o f collagenase
exerted b y serum m a c r o g l o b u l i n s . B i r k e d a l - H a n s e n , et 39
al.,
have noted the same phenomenon w i t h respect to
increased
collagenase
activity when
the supernatant
f r o m macrophage cultures a n d bovine gingivae in vitro was exposed to trypsin a n d / o r extracts o f mast cell preparations. It is possible that mast cell proteases re lease the α
2
macroglobulin-collagenase c o m p l e x a n d
thereby contribute to an increase i n collagenolytic activ ity. SUMMARY
L i g h t m i c r o s c o p i c e x a m i n a t i o n o f the mast cell d i s t r i bution i n various o r a l tissues i n germ-free a n d conven t i o n a l beagle dogs revealed no differences between the two animals. M a s t cells were observed i n a l l tissues e x a m i n e d , with an increasing order o f incidence as follows: cervical l y m p h node, p a r o t i d and submandibular glands, m a r g i n a l gingivae, b u c c a l mucosa and the middle one-third o f the tongue. A l l four types (round, oval, elongated, a n d pseudopodial) o f mast cells cell shapes previously described i n the literature were observed. T h e m a r g i n a l gingivae associated w i t h the m a n d i b u l a r , sec ond bicuspids was e x a m i n e d by light m i c r o s c o p y and was classified as to degree o f i n f l a m m a t o r y involvement. In a c o m p a r i s o n o f germ-free a n d conventional animals, the d i s t r i b u t i o n o f the degree o f i n f l a m m a t i o n proved to be r a n d o m . C o r r e l a t i o n between mast cell densities and the degree o f i n f l a m m a t o r y infiltration was not statistically significant. H o w e v e r , mast cells tended to decrease as the i n f l a m m a t i o n became more severe. E l e c t r o n m i c r o s c o p i c e x a m i n a t i o n o f mast cells i n the buccal mucosa revealed no difference i n m o r p h o l o g y o f cells from germ-free o r c o n v e n t i o n a l a n i m a l s . C y t o p l a s m i c granules were o f two basic types, one exhibited an amorphous m a t r i x o f u n i f o r m density a n d the other consisted o f laminated coils o f various sizes and densities. REFERENCES
1. Ehrlich, P.: Beitrage zur Kenntnis der Anilinfarbungen und ihrer Verwendung in der mikroskopischen Technik. Arch
Mikros Anat 13: 263, 1877. 2. Recklinghausen, F . von: Ueber Eiter- und Bindegewebskttrperchen. Virchow Arch Pathol Anat 28: 157, 1863. 3. Waldeyer, W.: Ueber Bindegewebszellen. Arch Mikros Anat 11: 176, 1875. 4. Unna, P. G . : Histopathologie der Hautkrankheiten {Neurofibrom, Urticaria pigmentosa, milaria, syphilide). Leo pold Voss, Hamburg, 1894. 5. Watson, M . L . : Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol 4: 475, 1958. 6. Reynolds, E. S.: The use of lead citrate at high p H as an electron opaque stain in electron microscopy. J Biophys Biochem Cytol 17: 208, 1963. 7. Zachrisson, B. V . : Mast cells of the human gingivae. II. Metachromatic cells at low p H in healthy and inflammed tissue. J Periodont Res 2: 87, 1967. 8. Jorpes, E., Holmgren, H . , and Wilander, O.: Uber das Vorkommen von Heparin in den Gefasswanden und in den Augen. Z Mikrosk Anat Forsch 42: 279, 1937, cited by Keller, R.: Monographs in Allergy. II. Tissue mast cells in immune reactions. New York, American Elsevier C o . , 1966. 9. Robinson, L . P., and De Marco, T . J . : Alteration of mast cell densities in experimentally inflamed human gingivae. J Periodontol 43: 614, 1972. 10. Tanaka, Y . , and Goodman, J . R.: Electron Microscopy of Human Blood Cells. New Y o r k , Harper & Row, Publishers, 1972. 11. Listgarten, M . A . , and Heneghan, J . B.: Chronic inflammation in the gingival tissue of germfree dogs. Arch Oral Biol 16: 1207, 1971. 12. Listgarten, M . A . , and Heneghan, J . B.: Observations on the periodontium and acquired pellicle of adult germfree dogs. J Periodontol 44: 85, 1973. 13. Gomori, G . : Distribution of acid phosphatase in the tissues under normal and under pathologic conditions. Arch Pathol 32: 189, 1941. 14. Pioch, W.: Uber die Darstellung saurer Mucopolysac charide mit dem Kupferphthalocyaninfarbstoff Astrablau. Vir chow Arch Pathol Anat 330: 337, 1957. 15. Fullmer, H . M . : Differences in mechanism in staining reactions for mast cells. Nature (London) 183: 1274, 1959. 16. Lison, L . : Etudes sur la metachromasie, colorants metachromatiques et substances chromotropes. Arch Biol (Paris) 46: 599, 1935. 17. Riley, J . R.: The relationship of the tissue mast cell to the blood vessels in the rat. J. Pathol Bad (London) 65: 461, 1953. 18. Hibbs, R. G . , Burch, G . E., and Phillips, J . H . : Electron microscopic observations on the human mast cell. Am Heart J 60: 121, 1960. 19. Angelopoulus, A . P.: Studies of mast cells in the human gingiva. I. Morphology. J Periodont Res 8: 28, 1973. 20. Weinstock, A . , and Albright, J . T.: The fine structure of mast cells in normal human gingiva. J Ultrastruct Res 17: 245, 1967. 21. Barnett, M . L . : The fine structure of human epithelial mast cells in periodontal disease. J Periodont Res 8: 371, 1973. 22. Barnett, M . L . : The fine structure of human connective tissue mast cells in periodontal disease. J Periodont Res 9: 84, 1974. 23. Mota, I., Ferri, A . G . , and Junqueira, L . C : Action of peptone on the mast cells and histamine content of dog tissues. Acta Haemwtol (Basel) 15: 409, 1956. 24. Arvy, L . and Quivy, D.: Donnees sur la repartition des labrocytes chez le chien. Compes Rend Assoc Anat 42: 234, 1955. 25. Todaro, F.: Morfologia, frequenza ed ubicazione degli elementi granulosi basofilli del connettivo nei territori dentali, paradentali e buccali dell'uomo e dei mammiferi. Stomatologia
Volume 47 Number 4
Mast Cell
37: 438, 1939. 26. A r v y , L . : Les ganglions lymphatiques et les liberateurs d'histamine. Compt Rens Assoc Anat 44: 107, 1958. 27. Carranza, F . A . and Cabrini, R . L . : Mast cells in human gingiva. Oral Surg 8: 1093, 1955. 28. Zachrisson, B. U . : Mast cells of the human gingiva. I V . Experimental gingivitis. J Periodont Res 4: 46, 1968. 29. Shelton, L . E . , and H a l l , W . B.: Human gingival mast cells. Effects of chronic inflammation. J Periodont Res 2: 214, 1968. 30. Shapiro, S., Ulmansky, M . , and Scheuer, M . : Mast cell population in gingiva affected by chronic destructive periodon tal disease. J Periodontol 40: 276, 1969. 31. Zachrisson, B. U . : The role of mast cells in gingivitis. Periodontologie 23: 151, 1969. 32. Angelopoulus, A . P.: Studies of mast cells in the human gingiva. II. Topographical distribution. J Periodon Res 8: 314,
Distribution
235
1973. 33. Dolby, A . E., and Allison, R . T.: Quantitative changes in the mast cell population in M i k u l i c z ' s recurrent oral aphthae. J Dent Res 48: 901, 1969. 34. H a l l , W . B.: Mast cells in desquamative gingivitis, lichen planus, and pemphigoid. Oral Surg 28: 646, 1969. 35. Jandinski, J . J . , Sonis, S., and Doku, H . C : The incidence of mast cells in selected oral lesions. Oral Surg 34: 245, 1972. 36. Lagnunoff, D . : The properties of mast cell proteases. Biochem Pharmacol (Suppl.) 221, 1968. 37. Simpson, J . W . , and Taylor, C : Regulation of gingival collagenase: A possible role for a mast cell factor. Proc Soc Exp Biol Med 145: 42, 1974. 39. Birkedal-Hansen, H . , Cobb, C . M . , Taylor, R . E . , and Fullmer, H . M . : Activation of latent macrophage and bovine gingival collagenase activity. In Press.
Abstracts INTRAORAL PSORIASIS VULGARIS
Rudolph, R. I., and Rudolph. L . P . Int J Dermatol 14: 101, March, 1975. Oral lesions of psoriosis are very rare with percentages given in the literature varying from 2% of 100 patients examined to as few as 1% of 500 patients. Following an historic review, the many clinical variations were described. The lesions may be single or multiple, large or small, white or yellow or even pigmented usually surrounded by an erythema. The lesions were not diagnostic as they resemble such conditions as syphilis, lichen planus, or even the rare dyskeratosis congenita. A biopsy was essential to distinguish from leukoplakia. Leukoplakia has dyskeratosis and horn pearl formation, not found in the psoriasis. To make the correct diagnosis, criteria which have been described include the location of the lesion, an associated cutaneous lesion, and the results of a biopsy. Department of Dermatology, Hospital of the University of Pennsylvania, Duhring Laboratories, 3600 Spruce Street, Philadelphia, Pennsylvania 19104
PERIODONTAL C H A N G E S IN DISTANT T E E T H D U E TO T R A U M A FROM O C C L U S I O N
Nascimento, A., and Sallum, A. W. J Periodont Res 10: 44, February, 1975. Eight marmosets were subjected to modification of occlusal relation ships in the maxillary molar regions using orthodontic wire cemented into occlusal perforations. Periodontal changes in the lower canine and first premolar area were studied over period of 4 weeks. Two animals were sacrificed at 7-day intervals. It was shown that the teeth were driven mesially producing detectable histological changes in the sup porting periodontal tissues of the canine and first premolar. On the pressure side, mesial to the premolar, there was resorption of bone, and on the tension side, distal to the canine, there was newly formed bone. Faculdade de Odontologica de Piracicaba, UNICAMP, Rua D. Pedro II, No. 627 cx Postal 52, 13.400-Piracicaba, Sao Paulo, Brazil
E F F E C T OF POSTMORTEM TISSUE FIXATION O N T O O T H MOBILITY A N D T H E E F F E C T OF C H E W I N G A P P L E S O N D E N T A L P L A Q U E A N D F O O D DEBRIS
Birkeland, J. M . , and Jorkjend, L . Comm Dent Oral Epidemiol 2: 161, No. 4, 1974. The present study was conducted to assess a possible cleansing effect of apple chewing on the amount of plaque in children. Of the 47 12-year-old children chosen, 24 children were asked to eat an unpeeled apple and then to rinse the mouth with tap water, immediately after which the plaque was rescored. This evaluation revealed that chewing apples had no cleansing effect on moderate amounts of plaque. In the second part, 27 children ate a bun each, then an apple; while 30 children had only the bun. This examination showed that chewing apples removed food debris and accelerated the oral sugar clearance. Dental Institute of Experimental Research, University of Oslo, Blindern, Oslo 3, Norway
P O C K E T D E P T H IN H U M A N BEINGS
Gillette, W. B . Oral Surg 39: 130, January, 1975. Pre- and postexamination of basic periodontal disease features was made of a patient with a confirmed malignant lesion in the floor of the mouth prior to resection of the mandible for the purpose of determining whether or not changes occurred in mobility and pocket depths following fixation. It was demonstrated that the mobilities 4 days postfixation, were markedly reduced. Changes in pocket depths were seen both to increase and decrease by an average of 1.2 mm. Of the 44 locations, however, 29 remained the same over a 34-day period postfixation. It was concluded that it may be possible to extrapolate pocket depths on postmortem studies but mobilities would be unrelia ble. Veterans Administration Hospital, 1481 West 10th Street, Indian apolis, Indiana 46202
received suitable v i t a m i n and m i n e r a l supplements and continued sterility o f the germ-free a n i m a l s was deter m i n e d by weekly cultures o f fecal swabs and s e r o l o g i c a l assays o f serum from c o m m o n canine v i r a l antigens. A l l animals were sacrificed between the ages o f 10 to 12 months, with the exception o f one c o n v e n t i o n a l a n i m a l sacrificed at the age of 14 months. A l l germ-free a n i m a l s remained sterile u n t i l time o f death.
by
Immediately after
sacrifice,
the j a w s and
selected
tissue biopsies were fixed i n phosphate buffered f o r m a l i n
CHARLES M . COBB*
( p H 7.4) for a p p r o x i m a t e l y 12 days. A f t e r fixation, b l o c k
JAMES B . H E N E G H A N I
sections o f the right and left second m a n d i b u l a r bicus
DENIS M . L E B L A N C ‡
pids were processed for study by light m i c r o s c o p y . O t h e r tissues chosen for study included sections from
M I C H A E L J . DAVIS‡
the b u c c a l m u c o s a , cervical l y m p h node, s u b m a n d i b u l a r and p a r o t i d glands, and the m i d d l e one-third o f the
EHRLICH
1
tongue. A l l tissue was embedded i n paraffin and sec
is generally credited w i t h being the first to
tioned at 6 µ. A l t e r n a t e sections were stained respec
describe the mast cell. H o w e v e r , the granulated cells, known
as
mast
Recklinghausen
2
cells,
were
actually
described
tively with hematoxylin-eosin and with
by
1%
aqueous
toluidine blue.
3
and W a l d e y e r before E h r l i c h defined
them by their m e t a c h r o m a t i c granules. T h e term " m a s t
Biopsies of buccal m u c o s a and m a r g i n a l gingivae from
c e l l " originated from the overladen granules, which give
one germ-free and one conventional d o g were processed
the cells the appearance o f being overfed. In 1894, U n n a
for routine e x a m i n a t i o n by electron m i c r o s c o p y . C o l d
4
phosphate buffered gluteraldehyde was the i n i t i a l fixative
described a mastocytosis o f s k i n in u r t i c a r i a pigmentosa; since then significant i n f l a m m a t o r y , h e m o d y n a m i c , and
and cold 2% o s m i u m tetraoxide was used for postfixa-
hemostatic activities o f mast cells have been extensively
tion. T h e tissue was dehydrated in a graded series o f
explored. Despite a history that spans a century o f study,
alcohols and embedded in E p o n b l o c k s . T h i n sections
the b i o l o g i c functions o f the mast cell r e m a i n relatively
were
obscure.
acetate.
double
stained
with
lead
citrate
and
uranyl
5,6
It is widely accepted that the host response to bacterial
M a s t cells were counted at a m a g n i f i c a t i o n o f x 250
plaque a n d / o r plaque products is i n f l a m m a t o r y and that
using a grid that bounded an area o f 0.3 m m x 0.3 m m
this response is the major destructive factor i n perio
(0.9 m m ) . S i x sections were used from each tissue biopsy
d o n t a l disease. T h e m a n y c h e m i c a l mediators o f the
from each a n i m a l , and 12 fields were counted in each
2
i n f l a m m a t o r y process associated w i t h mast cell p o p u l a
section. C o u n t i n g of overlapping fields was avoided and
tions i n o r a l tissues have led to numerous investigations
only those cells that were u n m i s t a k a b l y identifiable as mast cells were counted. C e l l counts were converted to
and an extensive literature.
the number o f cells per square m i l l i m e t e r o f tissue.
T h e present report was m o t i v a t e d by the lack o f infor
A second evaluation was made o f the m a r g i n a l g i n
m a t i o n c o n c e r n i n g mast cell populations i n various o r a l tissues o f germfree animals, p a r t i c u l a r l y i n the beagle
givae associated with the
d o g , w h i c h is susceptible to p e r i o d o n t a l disease in the
bicuspids. T h e gingivae were graded as to the degree o f
m a n d i b u l a r right and
left
c o n v e n t i o n a l e n v i r o n m e n t and has become a m u c h used
i n f l a m m a t o r y infiltration using a scale o f slight, moder ate and severe. M a s t cell counts were taken from the area
e x p e r i m e n t a l m o d e l in p e r i o d o n t a l research.
just below the epithelium. A s i n the i n i t i a l cell counts, 12 MATERIALS AND METHODS
fields were counted in six tissue sections from each tooth from each a n i m a l . T h e collected data were evaluated for
S i x germ-free beagles (obtained by C a e s a r i a n deliv ery) and four c o n v e n t i o n a l a n i m a l s were acquired from the L o u i s i a n a State U n i v e r s i t y M e d i c a l C e n t e r , N e w O r l e a n s , L o u i s i a n a . T h e germ-free a n i m a l s were raised on a diet o f sterilized (autoclaved) B o r d o n ' s E s b i l a c P u r i n a D o g C h o w and water ad libitum. T h e conven t i o n a l a n i m a l s , w h i c h served as controls for this study, were fed the same diet i n nonsterile f o r m . A l l animals
differences between means by using the Student's t Test. A n a l y s i s for statistical significance c o m p a r e d the means within i n f l a m m a t o r y groups, i.e., slight vs. moderate, slight vs. severe, and moderate vs. severe. RESULTS
E x a m i n a t i o n o f mast cells in a l l tissues by light microscopy revealed the presence o f four basic cell morphologies: r o u n d , o v o i d , spindle or elongated, and pseudopodial. A l l four cell forms have been described in the l i t e r a t u r e . A s previously noted by R o b i n s o n and D e M a r c o , the density o f the connective tissue appeared to
* Department of Periodontics, University of Alabama-Birmingham, University Station, Birmingham, Alabama, 35294. † Louisiana State University, Medical Center, New Orleans, La 70112. ‡ 3320 North Hullen Street, Metairie, La 70002.
7-9
9
230
Volume 47 Number 4
Mast Cell Distribution
determine w h i c h o f the four basic cell forms was p r e d o m i n a n t in any given tissue. T h e elongated cell forms were more prevalent in dense connective tissue whereas rounded cell forms were preponderant in loose connective tissue areas ( F i g . 1 and inset). U l t r a s t r u c t u r a l e x a m i n a t i o n revealed mast cells to have the characteristic, numerous projections from the cell m e m b r a n e , pseudopodia, and m i c r o v i l l i , which are seldom seen i n other granulated cell t y p e s . E a c h cell observed contained a single nucleus located in an eccen tric position. T h e usually irregular nuclear m a r g i n was a result o f adjacent c y t o p l a s m i c granules ( F i g . 3). F e w 10
231
G o l g i structures were noted and the e n d o p l a s m i c reticu l u m were also sparse. T h e c y t o p l a s m contained numer ous fibrillar structures ( F i g . 2), diffusely distributed ribosomes and scattered glycogen granules. M a s t cells contained specific granules that varied from those o f homogeneous density ( F i g . 3) to those with coils and l a m i n a t e d structures ( F i g . 2). T h e latter structures rep resented a well-organized a r r a y o f p a r a l l e l , alternate dense and less dense lines with a periodicity o f a p p r o x i mately 75 to 100 A. T a b l e 1 presents the quantitative d i s t r i b u t i o n o f mast cells in the various o r a l tissues o f both germ-free and
FIGURE 1. Sections (1 n thick) stained with toluidine blue demonstrating mast cell population in loose connective tissue of the buccal mucosa from a germ-free beagle dog. Note perivascular location (V) and fact that most cells represent the round and/or oval morphology classification (insert). (Original magnification 250 x , insert 800 x . )
FIGURE 2. A section through the cytoplasm of a mast cell from the buccal mucosa of a germ-free beagle dog. The cytoplasm is densely packed with a heterogenous population of granules exhibiting coiled and laminated internal structure. Note the numerous cytoplasmic vacuoles (V) and fine filaments (arrows). (Original magnification 30,000 x . )
232
J. Periodontol. April, 1976
Cobb, Heneghan, LeBlanc, Davis
F I G U R E 3. A section through the cytoplasm of a mast cell from the buccal mucosa of a conventional beagle dog. The cytoplasm exhibits many membrane bound granules (arrows), each with an amorphous matrix of uniform density. Several cytoplasmic vacuoles were observed and occasionally a lipid droplet (L). Note also how the nuclear membrane ( N M ) presents an irregular contour due to proximity of the granules. (Original magnification 33,000 x.)
2
TABLE 1. Mast Cell Counts (cells/mm ) and Standard Deviations (S.D.) in Various Oral Tissues of Germfree vs. Conventional Beagle Dogs Germfree Cells/mm
2
Conventional
S.D.
Cells/mm
2
S.D.
Tongue Buccal Mucosa
669.3 449.0
±126.5 ±110.2
598.9 487.0
±111.2 ±128.2
Marginal Gingiva
305.5 88.4
±80.8 ±33.3
324.0 101.7
±91.0 ±37.6
106.0
±35.9
73.7
±19.9
±9.9
51.7
±12.8
Parotid Gland Submandibular Gland Cervical Lymph
38.1
Node * All values represent means ± S.D.
c o n v e n t i o n a l a n i m a l s . It should be noted that the counts obtained for s i m i l a r tissues from both types o f a n i m a l s were not significantly different. T h e mast cell p o p u l a t i o n o f the tongue was evenly distributed between the connective tissue adjacent to surface e p i t h e l i u m a n d that associated with the tongue musculature. M a s t cells o f the salivary glands were confined to the connective tissue s t r o m a and were usually adjacent to c i r c u l a t o r y vessels. In b u c c a l m u c o s a a n d m a r g i n a l gingivae the mast cell populations were d i s t r i b uted throughout the l a m i n a p r o p r i a with no p a r t i c u l a r pattern o f c o n c e n t r a t i o n . In contrast, cervical l y m p h nodes c o n t a i n e d relatively few mast cells a n d most were concentrated at the periphery o f the node and adjacent to the connective tissue capsule. E x a m i n a t i o n o f 20 teeth with their m a r g i n a l gingivae (from six germ-free a n d four conventional a n i m a l s )
F I G U R E 4. Example of degranulating mast cell from severely inflamed marginal gingiva of a germ-free beagle dog. A I µ-thick section from Epon embedded tissue stained with toluidine blue. (Original magnification 800 x.)
showed the d i s t r i b u t i o n o f the three i n f l a m m a t o r y groups to be o f r a n d o m pattern. T h u s , no c o n c l u s i o n c o u l d be made with respect to degree o f i n f l a m m a t i o n o f m a r g i n a l gingivae i n germ-free vs. the c o n v e n t i o n a l beagle d o g . N o a n i m a l was completely free o f i n f l a m m a t i o n at the m i c r o s c o p i c level, c o i n c i d i n g with a previous observation made by L i s t g a r t e n a n d H e n e g h a n , w h o used a n i m a l s from this same germ-free c o l o n y . T h e m a r g i n a l gingivae o f three teeth exhibited severe i n f l a m m a t o r y infiltration as well as p r o n o u n c e d food a n d / o r hair i m p a c t i o n . A l t h o u g h not a quantitative observation, it was noted that i n the severe i n f l a m m a t i o n group, m a n y of the mast cells appeared to be degranulating ( F i g . 4). T h i s was not true o f mast cells associated w i t h the slight and moderate i n f l a m m a t o r y groups. T a b l e 2 presents the 1 1 , 1 2
Volume 47 Number 4
Mast Cell Distribution
233
2
TABLE 2. Mast Cell Counts of Marginal Gingivae According to Inflammatory Status {cells/mm ) Inflammatory Groups Slight
Moderate 11
6
Number of specimens in
Severe 3
group 3154
1993
Total number of mast cells
388
counted Mean count Standard deviation Range of count Statistical significance
332.17
286.73
129.33
+58.95
+46.85
+92.52
206-421
148-406
Slight vs. moderate
Moderate vs. severe
Severe vs. slight
None
None
None
results o f data analysis concerning the means o f mast cell counts o f m a r g i n a l gingivae a c c o r d i n g to i n f l a m m a t o r y status. N o t e that the difference between the means and, therefore, the difference i n n u m b e r o f mast cells is not significant when any t w o groups are considered. T h e o v e r a l l tendency was for a decrease i n the number o f mast cells as i n f l a m m a t i o n increased i n severity. DISCUSSION
M a s t cells, w h i c h are inherent i n the connective tissue o f m a n y organ systems, routinely exhibit a perivascular l o c a t i o n , an association thought to favor release o f their g r a n u l a r contents into the c i r c u l a t o r y system. B a s i c aniline dyes, such as methylene blue, toluidine blue o r cresyl violet are classically used to demonstrate the presence o f mast cells. These dyes elicit a m e t a c h r o m a s i a of specific c y t o p l a s m i c granules, a characteristic that stems from the strong negative charge o f the sulfated carbohydrate components o f the granule contents. T h e positive staining o f granules by the P A S reaction, alcian blue, astra b l u e , or colloidal i r o n suggests the presence o f acid m u c o p o l y s a c c h a r i d e s . These h i s t o c h e m i c a l considerations can be explained by the large a m o u n t of heparin i n the g r a n u l e s . T h e difference i n staining reactions o f mast cells w i t h i n the same tissue a n d / o r between tissues suggests either a heterogeneity o f c h e m i c a l constituents o r a difference i n cell m a t u r a t i o n . 13
14
1 5
16
17
In this study, the light a n d electron m i c r o s c o p i c observations o f mast cells indicate that the general m o r p h o l o g y o f this cell type, i n germ-free a n d conven t i o n a l beagle dogs, resembles that found i n m a n by other investigators. T h e four basic cell shapes described in the l i t e r a t u r e , as well as s i m i l a r u l t r a s t r u c t u r a l characteristics, were observed i n tissues o f both g e r m free a n d c o n v e n t i o n a l a n i m a l s . T w o basic granule m o r phologies were observed: one type presented an a m o r phous m a t r i x o f u n i f o r m density, a second type exhibited l a m i n a t e d coils o f various sizes a n d densities. T h e t w o types o f granules were present i n both elongated and o v a l shaped cells. S i m i l a r observations have been reported i n studies o f the specific granules i n mast cells o f other animal species. 9 , 1 8 , 1 9
7 , 8
20,22
In this study, the d i s t r i b u t i o n o f mast cells i n various
99-156
tissues (Table 1) agrees with the investigations o f M o t a , et a l . , A r v e y , et al., and T o d a r o , who used dogs as the e x p e r i m e n t a l a n i m a l . In most a n i m a l species, the tongue and b u c c a l m u c o s a appear to be relatively rich i n mast cells, whereas salivary glands a n d l y m p h nodes exhibit scant p o p u l a t i o n s . In this respect, the g e r m free beagles o f this study showed no v a r i a t i o n from what is currently accepted as the n o r m a l pattern o f d i s t r i b u tion. 23
24
2 5
2 3 l 2 6
Because mast cells are a n o r m a l cellular component o f healthy gingivae, both a n i m a l a n d h u m a n , nu merous investigations have sought to correlate the n u m ber o f mast cells with the degree o f i n f l a m m a t o r y involvement. T h e conflicting results produced by these studies is due, i n part, to differences i n histologic technique and a lack o f adequate controls. T h e i m p r o v e d techniques a n d m o r e precise controls used i n recent investigations have yielded m o r e consistent results. These results demonstrate that the number o f mast cells decreases d u r i n g acute i n f l a m m a t i o n , possibly because degranulation renders them undetectable by the usual histologic s t a i n s . I n contrast, the mast cell p o p u l a t i o n in areas o f c h r o n i c i n f l a m m a t i o n exceeds that i n healthy g i n g i v a e . In this study, the tendency for a decrease o f mast cells that accompanies increased i n f l a m m a t i o n m a y be explained b y a c o m b i n a t i o n o f t w o factors: (a) m a r g i n a l gingivae o f three teeth exhibiting the most severe i n f l a m m a t o r y infiltration were also characterized by the acute phase o f the i n f l a m m a t o r y reaction; (b) only cells definitely identified as mast cells were counted, thus faintly stained cells were not included i n the t o t a l count. In his study, Z a c h r i s s o n counted lightly stained cells, considering them as i m m a t u r e mast cells, which explains his finding o f an increase i n mast cell p o p u l a t i o n d u r i n g c h r o n i c g i n g i v a l i n f l a m m a t i o n . A s regards acute i n f l a m m a t i o n , A n g e l o p o u l o s has stated that few mast cells are found i n areas infiltrated by p o l y m o r p h o n u c l e a r leukocytes. 2 3 l 2 5 , 2 7 - 2 9
27,30
31
31
3 1
3 2
Besides being i m p l i c a t e d i n gingivitis, mast cells m a y contribute t o development o f lesions o f other o r a l m u c o u s membranes. U p o n finding that the mast cell p o p u l a t i o n was significantly reduced 48 hours after the c l i n i c a l onset o f M i k u l i c z ' s recurrent o r a l aphthae,
234
J. Periodontol. April, 1976
Cobb, Heneghan, LeBlanc, Davis
Dolby and A l l i s o n
3 3
suggested that the reduction m a y
represent a T y p e I V allergic reaction. H a l l
3 4
reported a
seemingly specific pattern o f mast cell d i s t r i b u t i o n i n the following
mucous
membrane
diseases:
lichen planus,
p e m p h i g o i d , a n d desquamative gingivitis. J a n d i n s k i , et 35
al.,
suggested that the increased number o f mast cells
present i n low-grade c a r c i n o m a m a y result from an i m m u n e response t o antigenic s t i m u l a t i o n . A s yet, the precise role o f the mast cell i n i n f l a m m a t i o n has not been elucidated. H o w e v e r , several good reviews o f the possible b i o l o g i c functions have published.
3 6 , 3 7
been
A recent finding, the synergistic role o f
the mast cell a n d an increased activity o f m a m m a l i a n collagenase, deserves further investigation a n d m a y be found to account for the l o c a l i z e d collagenolytic activity in p e r i o d o n t a l pocket f o r m a t i o n . S i m p s o n and T a y l o r
3 8
reported that the in vivo c o n t r o l o f collagenase is, i n part, a factor o f e n z y m a t i c function a n d that the enzymes i n v o l v e d are elaborated b y mast cells. These mast cell factors/enzymes
reversed the i n h i b i t i o n o f collagenase
exerted b y serum m a c r o g l o b u l i n s . B i r k e d a l - H a n s e n , et 39
al.,
have noted the same phenomenon w i t h respect to
increased
collagenase
activity when
the supernatant
f r o m macrophage cultures a n d bovine gingivae in vitro was exposed to trypsin a n d / o r extracts o f mast cell preparations. It is possible that mast cell proteases re lease the α
2
macroglobulin-collagenase c o m p l e x a n d
thereby contribute to an increase i n collagenolytic activ ity. SUMMARY
L i g h t m i c r o s c o p i c e x a m i n a t i o n o f the mast cell d i s t r i bution i n various o r a l tissues i n germ-free a n d conven t i o n a l beagle dogs revealed no differences between the two animals. M a s t cells were observed i n a l l tissues e x a m i n e d , with an increasing order o f incidence as follows: cervical l y m p h node, p a r o t i d and submandibular glands, m a r g i n a l gingivae, b u c c a l mucosa and the middle one-third o f the tongue. A l l four types (round, oval, elongated, a n d pseudopodial) o f mast cells cell shapes previously described i n the literature were observed. T h e m a r g i n a l gingivae associated w i t h the m a n d i b u l a r , sec ond bicuspids was e x a m i n e d by light m i c r o s c o p y and was classified as to degree o f i n f l a m m a t o r y involvement. In a c o m p a r i s o n o f germ-free a n d conventional animals, the d i s t r i b u t i o n o f the degree o f i n f l a m m a t i o n proved to be r a n d o m . C o r r e l a t i o n between mast cell densities and the degree o f i n f l a m m a t o r y infiltration was not statistically significant. H o w e v e r , mast cells tended to decrease as the i n f l a m m a t i o n became more severe. E l e c t r o n m i c r o s c o p i c e x a m i n a t i o n o f mast cells i n the buccal mucosa revealed no difference i n m o r p h o l o g y o f cells from germ-free o r c o n v e n t i o n a l a n i m a l s . C y t o p l a s m i c granules were o f two basic types, one exhibited an amorphous m a t r i x o f u n i f o r m density a n d the other consisted o f laminated coils o f various sizes and densities. REFERENCES
1. Ehrlich, P.: Beitrage zur Kenntnis der Anilinfarbungen und ihrer Verwendung in der mikroskopischen Technik. Arch
Mikros Anat 13: 263, 1877. 2. Recklinghausen, F . von: Ueber Eiter- und Bindegewebskttrperchen. Virchow Arch Pathol Anat 28: 157, 1863. 3. Waldeyer, W.: Ueber Bindegewebszellen. Arch Mikros Anat 11: 176, 1875. 4. Unna, P. G . : Histopathologie der Hautkrankheiten {Neurofibrom, Urticaria pigmentosa, milaria, syphilide). Leo pold Voss, Hamburg, 1894. 5. Watson, M . L . : Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol 4: 475, 1958. 6. Reynolds, E. S.: The use of lead citrate at high p H as an electron opaque stain in electron microscopy. J Biophys Biochem Cytol 17: 208, 1963. 7. Zachrisson, B. V . : Mast cells of the human gingivae. II. Metachromatic cells at low p H in healthy and inflammed tissue. J Periodont Res 2: 87, 1967. 8. Jorpes, E., Holmgren, H . , and Wilander, O.: Uber das Vorkommen von Heparin in den Gefasswanden und in den Augen. Z Mikrosk Anat Forsch 42: 279, 1937, cited by Keller, R.: Monographs in Allergy. II. Tissue mast cells in immune reactions. New York, American Elsevier C o . , 1966. 9. Robinson, L . P., and De Marco, T . J . : Alteration of mast cell densities in experimentally inflamed human gingivae. J Periodontol 43: 614, 1972. 10. Tanaka, Y . , and Goodman, J . R.: Electron Microscopy of Human Blood Cells. New Y o r k , Harper & Row, Publishers, 1972. 11. Listgarten, M . A . , and Heneghan, J . B.: Chronic inflammation in the gingival tissue of germfree dogs. Arch Oral Biol 16: 1207, 1971. 12. Listgarten, M . A . , and Heneghan, J . B.: Observations on the periodontium and acquired pellicle of adult germfree dogs. J Periodontol 44: 85, 1973. 13. Gomori, G . : Distribution of acid phosphatase in the tissues under normal and under pathologic conditions. Arch Pathol 32: 189, 1941. 14. Pioch, W.: Uber die Darstellung saurer Mucopolysac charide mit dem Kupferphthalocyaninfarbstoff Astrablau. Vir chow Arch Pathol Anat 330: 337, 1957. 15. Fullmer, H . M . : Differences in mechanism in staining reactions for mast cells. Nature (London) 183: 1274, 1959. 16. Lison, L . : Etudes sur la metachromasie, colorants metachromatiques et substances chromotropes. Arch Biol (Paris) 46: 599, 1935. 17. Riley, J . R.: The relationship of the tissue mast cell to the blood vessels in the rat. J. Pathol Bad (London) 65: 461, 1953. 18. Hibbs, R. G . , Burch, G . E., and Phillips, J . H . : Electron microscopic observations on the human mast cell. Am Heart J 60: 121, 1960. 19. Angelopoulus, A . P.: Studies of mast cells in the human gingiva. I. Morphology. J Periodont Res 8: 28, 1973. 20. Weinstock, A . , and Albright, J . T.: The fine structure of mast cells in normal human gingiva. J Ultrastruct Res 17: 245, 1967. 21. Barnett, M . L . : The fine structure of human epithelial mast cells in periodontal disease. J Periodont Res 8: 371, 1973. 22. Barnett, M . L . : The fine structure of human connective tissue mast cells in periodontal disease. J Periodont Res 9: 84, 1974. 23. Mota, I., Ferri, A . G . , and Junqueira, L . C : Action of peptone on the mast cells and histamine content of dog tissues. Acta Haemwtol (Basel) 15: 409, 1956. 24. Arvy, L . and Quivy, D.: Donnees sur la repartition des labrocytes chez le chien. Compes Rend Assoc Anat 42: 234, 1955. 25. Todaro, F.: Morfologia, frequenza ed ubicazione degli elementi granulosi basofilli del connettivo nei territori dentali, paradentali e buccali dell'uomo e dei mammiferi. Stomatologia
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37: 438, 1939. 26. A r v y , L . : Les ganglions lymphatiques et les liberateurs d'histamine. Compt Rens Assoc Anat 44: 107, 1958. 27. Carranza, F . A . and Cabrini, R . L . : Mast cells in human gingiva. Oral Surg 8: 1093, 1955. 28. Zachrisson, B. U . : Mast cells of the human gingiva. I V . Experimental gingivitis. J Periodont Res 4: 46, 1968. 29. Shelton, L . E . , and H a l l , W . B.: Human gingival mast cells. Effects of chronic inflammation. J Periodont Res 2: 214, 1968. 30. Shapiro, S., Ulmansky, M . , and Scheuer, M . : Mast cell population in gingiva affected by chronic destructive periodon tal disease. J Periodontol 40: 276, 1969. 31. Zachrisson, B. U . : The role of mast cells in gingivitis. Periodontologie 23: 151, 1969. 32. Angelopoulus, A . P.: Studies of mast cells in the human gingiva. II. Topographical distribution. J Periodon Res 8: 314,
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1973. 33. Dolby, A . E., and Allison, R . T.: Quantitative changes in the mast cell population in M i k u l i c z ' s recurrent oral aphthae. J Dent Res 48: 901, 1969. 34. H a l l , W . B.: Mast cells in desquamative gingivitis, lichen planus, and pemphigoid. Oral Surg 28: 646, 1969. 35. Jandinski, J . J . , Sonis, S., and Doku, H . C : The incidence of mast cells in selected oral lesions. Oral Surg 34: 245, 1972. 36. Lagnunoff, D . : The properties of mast cell proteases. Biochem Pharmacol (Suppl.) 221, 1968. 37. Simpson, J . W . , and Taylor, C : Regulation of gingival collagenase: A possible role for a mast cell factor. Proc Soc Exp Biol Med 145: 42, 1974. 39. Birkedal-Hansen, H . , Cobb, C . M . , Taylor, R . E . , and Fullmer, H . M . : Activation of latent macrophage and bovine gingival collagenase activity. In Press.
Abstracts INTRAORAL PSORIASIS VULGARIS
Rudolph, R. I., and Rudolph. L . P . Int J Dermatol 14: 101, March, 1975. Oral lesions of psoriosis are very rare with percentages given in the literature varying from 2% of 100 patients examined to as few as 1% of 500 patients. Following an historic review, the many clinical variations were described. The lesions may be single or multiple, large or small, white or yellow or even pigmented usually surrounded by an erythema. The lesions were not diagnostic as they resemble such conditions as syphilis, lichen planus, or even the rare dyskeratosis congenita. A biopsy was essential to distinguish from leukoplakia. Leukoplakia has dyskeratosis and horn pearl formation, not found in the psoriasis. To make the correct diagnosis, criteria which have been described include the location of the lesion, an associated cutaneous lesion, and the results of a biopsy. Department of Dermatology, Hospital of the University of Pennsylvania, Duhring Laboratories, 3600 Spruce Street, Philadelphia, Pennsylvania 19104
PERIODONTAL C H A N G E S IN DISTANT T E E T H D U E TO T R A U M A FROM O C C L U S I O N
Nascimento, A., and Sallum, A. W. J Periodont Res 10: 44, February, 1975. Eight marmosets were subjected to modification of occlusal relation ships in the maxillary molar regions using orthodontic wire cemented into occlusal perforations. Periodontal changes in the lower canine and first premolar area were studied over period of 4 weeks. Two animals were sacrificed at 7-day intervals. It was shown that the teeth were driven mesially producing detectable histological changes in the sup porting periodontal tissues of the canine and first premolar. On the pressure side, mesial to the premolar, there was resorption of bone, and on the tension side, distal to the canine, there was newly formed bone. Faculdade de Odontologica de Piracicaba, UNICAMP, Rua D. Pedro II, No. 627 cx Postal 52, 13.400-Piracicaba, Sao Paulo, Brazil
E F F E C T OF POSTMORTEM TISSUE FIXATION O N T O O T H MOBILITY A N D T H E E F F E C T OF C H E W I N G A P P L E S O N D E N T A L P L A Q U E A N D F O O D DEBRIS
Birkeland, J. M . , and Jorkjend, L . Comm Dent Oral Epidemiol 2: 161, No. 4, 1974. The present study was conducted to assess a possible cleansing effect of apple chewing on the amount of plaque in children. Of the 47 12-year-old children chosen, 24 children were asked to eat an unpeeled apple and then to rinse the mouth with tap water, immediately after which the plaque was rescored. This evaluation revealed that chewing apples had no cleansing effect on moderate amounts of plaque. In the second part, 27 children ate a bun each, then an apple; while 30 children had only the bun. This examination showed that chewing apples removed food debris and accelerated the oral sugar clearance. Dental Institute of Experimental Research, University of Oslo, Blindern, Oslo 3, Norway
P O C K E T D E P T H IN H U M A N BEINGS
Gillette, W. B . Oral Surg 39: 130, January, 1975. Pre- and postexamination of basic periodontal disease features was made of a patient with a confirmed malignant lesion in the floor of the mouth prior to resection of the mandible for the purpose of determining whether or not changes occurred in mobility and pocket depths following fixation. It was demonstrated that the mobilities 4 days postfixation, were markedly reduced. Changes in pocket depths were seen both to increase and decrease by an average of 1.2 mm. Of the 44 locations, however, 29 remained the same over a 34-day period postfixation. It was concluded that it may be possible to extrapolate pocket depths on postmortem studies but mobilities would be unrelia ble. Veterans Administration Hospital, 1481 West 10th Street, Indian apolis, Indiana 46202
