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Glycogen in papillary hyperplasia of the palate
ORAL PATHOLOGY American Academy of Oral Pathology Donald Kerr, Editor
.
.
.
.
.
GLYCOGEN
.
.
.
.
IN PAPILLARY
Kendall Porter, D.D.X., MS., Kansas City, MO. Department of
Pathology,
University
.
.
.
.
.
.
HYPERPLASIA
.
School
.
OF THE
ana! Viota D. Flanagan,
of Eansas City,
.
H.T.
.
.
.
.
.
.
PALATE (ASCP),
B.S.,
of Dentistry
T
HE frequent occurrence of papillary hyperplasia of the palate has caused considerable interest, but etiological factors and pathologic significance have not yet been positively established. Although different lesions tend to display histologic similarity, the histochemical pattern is somewhat variable, making a definite correlation difficult at this time. The role of glycogen in the normal metabolism of tissues is well known. However, since rapidly growing cells are assumed to have greater carbohydrate requirements, investigations have been directed toward establishing whether or not cancer cells contain greater amounts of glycogen. It has been reported that less than half of the malignant tumors contain much, if any, g1ycogen.l Rut what of lesions that are classified as benign and yet have the potential, and in some instances the suggestion, of malignant transition? What is the histochemical nature of these tissues? Since papillary hyperplasia has been indicated as a premalignant lesion,*3 3 it would seem logical to investigate whether there is a histochemical correlation between this lesion and the true malignant tumors. This article will report the results of a glycogen study of lesions diagnosed clinically and histopathologically a.s papillary hyperplasia of the palate. MATERIALS
Fifteen distribution
AND
METHODS
cases of papillary hyperplasia were examined to determine the of glycogen. In four cases two separate biopsy specimens from
This study was supported by General Research SuPPort Grant l-GS-24, Sub-t, from the United States Public Health Service. National Institute of Dental Research, National Institutes of Health, Bethesda, Md. 1331
Fig. I.-A, Papillary hyperplasia showing inflmwrrmtion. cpithelial :md connective issue and Assures (F). Keratin is present at the crest of the papillae, and 3ara hypf ?rplasia, in the fissures. ~Hematoxylin anti eosin stain. Magnification, X100.) kera .tosis is present $3. Papillary hyperplasia stained to show the areas of keratin. (Mallory stain. M a Lgniflcati on, x1no.j
Volume 16 Number I I
GLYCOGEN
IN
PAPILLARY
1333
HYPERPLASIA
B. Fig. ,2.--A, Glycogen in papillary hyperplasia. ;;rry,gen 1s present at the crest of the papilla. B, ninutes. arrow).
Glycogen is most intense in fissures. Note the glycogen surrounding an epil
Control section showing the removal of glycogen after incubation in saliva There is a persistence of PAS-positive material in the center of the epithelial (Alcian blue--periodic acid-Schiff stain. Magnification. X100.)
f
The specimens wcrc fiscd in IO par cent neutral t’ol*malin. l’al*affill s~iions wcr(l cut and st.aincd by the following tnrthods : 1. Hematoxylin and eosin for diagnostic purposes~ for determining the status of the keratin layer and t,hc presence of inflammation, and for* general orim3t~ation (Fig. 1, J ) . 2. Mallory stain, to d&ermine keratinization (Fig. 1, H) . 3. Alcian blue-periodic acid-Schiff (AB-PAX) stain,‘ to determinc~ the glycogen content. Cont,rol studies were run on all AR-PAS sections. Sections wcw placed in saliva for 30 minutes after alcian-blue staining and then stained 1)~ 11~~2 PAS method. The glycogcn was removed in these sections (Fig. 2). The sections wcrc studied with the following r(‘lationships in tnind : 1. I’rcscnce or absence of glycogcn ‘L. Prcsencc of glycogen in certain specific parts of the cpithclium : A. Prickle-ccl1 la.yer and/or surface layer 73. ‘ ‘ Fissures ’ ’ ( ‘. Epithclial pearls I). Basal-ccl1 layer 3. Presence or absence of keratin and status of t,hc ktratin, if l)rcscnt : A. Mature keratinization I$. Incomplctc kcratinization C. I’arakerat,osis 4. Presence or abscncc of inflammation RESULTS
All of the spccimcns showed the prcsencc of glycogcn in the (~pithelium. In all instances, glycogcn was present in the prickle-cell layer (Fig. 2. ~1). In sixteen of the specimens, glycogen was present in “fissures” (Fig. 2, ~1). Ten of the specimens exhibited cpithelial pearl format,ion; glycogen was present, in these areas (Fig. 3). Two specimens showed t,he presence of glycogen in the basal layer (Fig. 4). Parakeratosis was seen in all but one of t,hc specimens. Some specimens showed a variation from incomplete keratinization to nonkeratinization in differcnt a.rcas of the same specimen. In all instances, glycogen was found in conjunction with a parakeratotic surface. Inflammation was present in all spccimens. The basement, membrane appcarctl to be intact in all specimens except in areas of intense inflammation, whrrc t,hcrc was evidence of liquefaction (Fig. 5). DISCUSSIOhT
Previous studies have reportecl the dist,ribution of glycogcn in normal and certain pathologic oral cpithelia. In most instances”-’ glycogcn was reported as being limited to the surface and/or prickle-cell layers and as being absent in
ULYCOGkX
IN
PAYlLLAHY Fig.
3.
Fig.
4.
1335
HYPEKYLd81A
Fig. 3.-Glycogen in epithelial pearls. The center area of PAS-positive, nonglycogen nlaterial (arrow). (Alcian blue-periodic flcation, X100.) Fig. I.-Glycogen present in the basal-cell layer (arrow). Schiff stain. Magnification, X100.)
the
is larger epithelial I ,earl acid-Schiff stain, Magni-
(Alcian
blue-periodic
acicl-
Dlasia. toxylin
Fig. B.-Intense inflammation in connective tissue and egitht:liurrr The basement membrane is indistinct because of the liquefaction and eosin stain. Ma,Kniflcation. x100.)
in
mpillars (ar~x~w I.
tIyper( Hema-
the basal-cell layer. In only one study was it reported in the basal-cell layer. Our findings concur in general with these previous findings. In two of our specimens, glycogen was present in the basal layer. The usual location was the prickle-cell layer. Glycogen was found to bc present when the surface was parakeratotic. Glycogen previously had been found to be related to parakcratosis.“? i Acanthosis was present in all specimens in relation to areas of glycogcn. In all instances, glycogen was found in the presence of inflammation, as no has been previously reported. 9*lo As all specimens showed inflammation, significance can be attached to this finding. Glycogen deposition was found to be heaviest in “fissured” areas, which lie between the papillae. It was in these areas that ratcuolization of individual cpithelial cells was noted in sections stain& with hematoxylin ant1 cosin. It has been suggestedl’ that these vacuoles may represc~nt ~lycogcn tloposition, unstained in hematoxylin sections. Our sections showed the glycogcn in these cells polarized in t,he typical “ glycogen flight” fixation arttlfact. One section showed a more even distribution of the glycogen (Fig. 6). Further studies are in progress, using freeze-substitution mf%hods in an attempt to localize the glycogen more accurately and show whether the vacuolated cells are, in fact, glycogen-laden ~11s. Our impression, at the motncnt. lcnns in this direction. No attempt has been made to interpret the results as far as the significance of the glycogen is concerned. Cahn, Eisenbud, and Blake” have stated that
Volume 16 Number I I
GLYCOQEN
IN
PAPILLARY
HYPERPLASlA
1337
B.
Fig. 6.--A,. Section of papillary hyperplasia showing greatly swollen cells and c,ells appearing devoid of cytoplasm. (Hematoxylin and eosin stain. Magnification, X100.) B, Section of papillary hyperplasia showing the “glycogen flight” fixation artefact ( ar TOW A) and the more even distribution of alyrogen in one area (arrow G). (Alcian blue-p6 3-i odic acid-Schiff stain. Magnification, X100.)
parakcratosis without glycogen may 1~ an ominous sign as I’ar as 1JretnaIignancy is concernecl. All of our specimens showed $ycogen and parakeratosis. Therefore, is papillary hyperplasia a significant prcmali~nant lesion ! Is papillary hyperplasia lradirqq to s~n;~a~ous-c~c~llcarcinoma only a. chance finclin~‘! This is still unrcsolvcd. ‘I%(’ prrscnt adiclc represents only one /?h>is(l of XII extcnsivo histochcmical stutly of this a~(1 other lesions 01’ the oral mn(:usu. COSCl,USIONS 7. Gllycogen is found where thcrc is parakeratosis, and it is absent. where there is hypcrkeratosis in papillary hyperplasia of the palate. 2. Acanthosis and inflammation arc seen in areas where there is alycogen deposition. Ko relationship is infrrretl from this stntly. 3. Glycogen is seen in the prickle-cell layer, in “fissures,” aronnd epithelial pearls, and, rarely. in thr basal cells of pa.pillary hypcrplasia of the palate. 4. In sonic instances, v;\.cuola.tctl c~l)ithclial ~11s caontaill heavy tlrposits of glycogen. REFERENCES 1. Smith, W., and Gault, S.: Essentials of Pathology, New York, 1942, D. ~~l’l)l(:toll-Cc!lturv Company, pp. 17-18. Lesions of the Oral Regions, D. 2. Robinson, H. B. G.: Neoplasms am1 “Precancerous” Clin. North America, pp. 621-626, November, 1957. Papillary Hpperplasla, J. Oral Surg., Anesth. 8r Hosp. T). 3. Waite, I).: Inflammatory Serv. 19: 210-214, 1961. 4. McManus, J. F. A., and Mowry, W.: Staining Methods. Histologic anll Histochemical, New York, 1960, Paul B. Hoeber, Inc., pp. 63-64. ljistribution of 5. Klingsherg, J., CaneelIaro, A., and Butcher, 0.: The Comparative and Acid Mucopolynaccharitles ill ljifferentGlycoge?, Succinic Dehydrogenase, aged Animals, J. D. Res. 40: 461-469, 1961. 6. Cahn, L., Eisenbud, L., and Blake, N.: Histochemieal Analysis of White Lesions of the Mouth. II. Analysis of Glpcogen Content, ORAL F~JRG., ORAL MED. k ORAL PATH.
15:
458-462,
1962.
7. Turesky, S., Glickman, I., and Provost, Process in Oral Lesions Diagnosed MED. & ORAL-PATH.
14: 442-453,
.J.: A Histochemieal Study of the Keratotic Clinically as Leukoplakia, ORAI. STW:., ORAT.
1961.
Stu(ly of the Attached 8. Turesky, S., Fisher, B., and Glickman, I.: A Histochemical Gingiva in Pregnancy, J. D. Res. 37: 1115-1122, 195s. !!. I)ewar, R.: Observations on the Composition and Metabolism of Normal ant1 Tnflametl Gingivae, J. Periodont. 26: 29-39, 1955. 10. Turesky, S., Glickman, I., and Litwin, R.: A Histochemical Evaluation of Normal and Inflamed Human Gingivae, J. D. Res. 30: 792-798, 1951. White Folded 1)ysplaaia of the I I. Zegarelli, E., Everett, F., and Kutscher, A.: Familial l\lucous Membranes, Oaar, SURG.,ORAr,M&D. & ORAL PATH. 14: 1436.1443, 1!161,
.
.
.
.
.
GLYCOGEN
.
.
.
.
IN PAPILLARY
Kendall Porter, D.D.X., MS., Kansas City, MO. Department of
Pathology,
University
.
.
.
.
.
.
HYPERPLASIA
.
School
.
OF THE
ana! Viota D. Flanagan,
of Eansas City,
.
H.T.
.
.
.
.
.
.
PALATE (ASCP),
B.S.,
of Dentistry
T
HE frequent occurrence of papillary hyperplasia of the palate has caused considerable interest, but etiological factors and pathologic significance have not yet been positively established. Although different lesions tend to display histologic similarity, the histochemical pattern is somewhat variable, making a definite correlation difficult at this time. The role of glycogen in the normal metabolism of tissues is well known. However, since rapidly growing cells are assumed to have greater carbohydrate requirements, investigations have been directed toward establishing whether or not cancer cells contain greater amounts of glycogen. It has been reported that less than half of the malignant tumors contain much, if any, g1ycogen.l Rut what of lesions that are classified as benign and yet have the potential, and in some instances the suggestion, of malignant transition? What is the histochemical nature of these tissues? Since papillary hyperplasia has been indicated as a premalignant lesion,*3 3 it would seem logical to investigate whether there is a histochemical correlation between this lesion and the true malignant tumors. This article will report the results of a glycogen study of lesions diagnosed clinically and histopathologically a.s papillary hyperplasia of the palate. MATERIALS
Fifteen distribution
AND
METHODS
cases of papillary hyperplasia were examined to determine the of glycogen. In four cases two separate biopsy specimens from
This study was supported by General Research SuPPort Grant l-GS-24, Sub-t, from the United States Public Health Service. National Institute of Dental Research, National Institutes of Health, Bethesda, Md. 1331
Fig. I.-A, Papillary hyperplasia showing inflmwrrmtion. cpithelial :md connective issue and Assures (F). Keratin is present at the crest of the papillae, and 3ara hypf ?rplasia, in the fissures. ~Hematoxylin anti eosin stain. Magnification, X100.) kera .tosis is present $3. Papillary hyperplasia stained to show the areas of keratin. (Mallory stain. M a Lgniflcati on, x1no.j
Volume 16 Number I I
GLYCOGEN
IN
PAPILLARY
1333
HYPERPLASIA
B. Fig. ,2.--A, Glycogen in papillary hyperplasia. ;;rry,gen 1s present at the crest of the papilla. B, ninutes. arrow).
Glycogen is most intense in fissures. Note the glycogen surrounding an epil
Control section showing the removal of glycogen after incubation in saliva There is a persistence of PAS-positive material in the center of the epithelial (Alcian blue--periodic acid-Schiff stain. Magnification. X100.)
f
The specimens wcrc fiscd in IO par cent neutral t’ol*malin. l’al*affill s~iions wcr(l cut and st.aincd by the following tnrthods : 1. Hematoxylin and eosin for diagnostic purposes~ for determining the status of the keratin layer and t,hc presence of inflammation, and for* general orim3t~ation (Fig. 1, J ) . 2. Mallory stain, to d&ermine keratinization (Fig. 1, H) . 3. Alcian blue-periodic acid-Schiff (AB-PAX) stain,‘ to determinc~ the glycogen content. Cont,rol studies were run on all AR-PAS sections. Sections wcw placed in saliva for 30 minutes after alcian-blue staining and then stained 1)~ 11~~2 PAS method. The glycogcn was removed in these sections (Fig. 2). The sections wcrc studied with the following r(‘lationships in tnind : 1. I’rcscnce or absence of glycogcn ‘L. Prcsencc of glycogen in certain specific parts of the cpithclium : A. Prickle-ccl1 la.yer and/or surface layer 73. ‘ ‘ Fissures ’ ’ ( ‘. Epithclial pearls I). Basal-ccl1 layer 3. Presence or absence of keratin and status of t,hc ktratin, if l)rcscnt : A. Mature keratinization I$. Incomplctc kcratinization C. I’arakerat,osis 4. Presence or abscncc of inflammation RESULTS
All of the spccimcns showed the prcsencc of glycogcn in the (~pithelium. In all instances, glycogcn was present in the prickle-cell layer (Fig. 2. ~1). In sixteen of the specimens, glycogen was present in “fissures” (Fig. 2, ~1). Ten of the specimens exhibited cpithelial pearl format,ion; glycogen was present, in these areas (Fig. 3). Two specimens showed t,he presence of glycogen in the basal layer (Fig. 4). Parakeratosis was seen in all but one of t,hc specimens. Some specimens showed a variation from incomplete keratinization to nonkeratinization in differcnt a.rcas of the same specimen. In all instances, glycogen was found in conjunction with a parakeratotic surface. Inflammation was present in all spccimens. The basement, membrane appcarctl to be intact in all specimens except in areas of intense inflammation, whrrc t,hcrc was evidence of liquefaction (Fig. 5). DISCUSSIOhT
Previous studies have reportecl the dist,ribution of glycogcn in normal and certain pathologic oral cpithelia. In most instances”-’ glycogcn was reported as being limited to the surface and/or prickle-cell layers and as being absent in
ULYCOGkX
IN
PAYlLLAHY Fig.
3.
Fig.
4.
1335
HYPEKYLd81A
Fig. 3.-Glycogen in epithelial pearls. The center area of PAS-positive, nonglycogen nlaterial (arrow). (Alcian blue-periodic flcation, X100.) Fig. I.-Glycogen present in the basal-cell layer (arrow). Schiff stain. Magnification, X100.)
the
is larger epithelial I ,earl acid-Schiff stain, Magni-
(Alcian
blue-periodic
acicl-
Dlasia. toxylin
Fig. B.-Intense inflammation in connective tissue and egitht:liurrr The basement membrane is indistinct because of the liquefaction and eosin stain. Ma,Kniflcation. x100.)
in
mpillars (ar~x~w I.
tIyper( Hema-
the basal-cell layer. In only one study was it reported in the basal-cell layer. Our findings concur in general with these previous findings. In two of our specimens, glycogen was present in the basal layer. The usual location was the prickle-cell layer. Glycogen was found to bc present when the surface was parakeratotic. Glycogen previously had been found to be related to parakcratosis.“? i Acanthosis was present in all specimens in relation to areas of glycogcn. In all instances, glycogen was found in the presence of inflammation, as no has been previously reported. 9*lo As all specimens showed inflammation, significance can be attached to this finding. Glycogen deposition was found to be heaviest in “fissured” areas, which lie between the papillae. It was in these areas that ratcuolization of individual cpithelial cells was noted in sections stain& with hematoxylin ant1 cosin. It has been suggestedl’ that these vacuoles may represc~nt ~lycogcn tloposition, unstained in hematoxylin sections. Our sections showed the glycogcn in these cells polarized in t,he typical “ glycogen flight” fixation arttlfact. One section showed a more even distribution of the glycogen (Fig. 6). Further studies are in progress, using freeze-substitution mf%hods in an attempt to localize the glycogen more accurately and show whether the vacuolated cells are, in fact, glycogen-laden ~11s. Our impression, at the motncnt. lcnns in this direction. No attempt has been made to interpret the results as far as the significance of the glycogen is concerned. Cahn, Eisenbud, and Blake” have stated that
Volume 16 Number I I
GLYCOQEN
IN
PAPILLARY
HYPERPLASlA
1337
B.
Fig. 6.--A,. Section of papillary hyperplasia showing greatly swollen cells and c,ells appearing devoid of cytoplasm. (Hematoxylin and eosin stain. Magnification, X100.) B, Section of papillary hyperplasia showing the “glycogen flight” fixation artefact ( ar TOW A) and the more even distribution of alyrogen in one area (arrow G). (Alcian blue-p6 3-i odic acid-Schiff stain. Magnification, X100.)
parakcratosis without glycogen may 1~ an ominous sign as I’ar as 1JretnaIignancy is concernecl. All of our specimens showed $ycogen and parakeratosis. Therefore, is papillary hyperplasia a significant prcmali~nant lesion ! Is papillary hyperplasia lradirqq to s~n;~a~ous-c~c~llcarcinoma only a. chance finclin~‘! This is still unrcsolvcd. ‘I%(’ prrscnt adiclc represents only one /?h>is(l of XII extcnsivo histochcmical stutly of this a~(1 other lesions 01’ the oral mn(:usu. COSCl,USIONS 7. Gllycogen is found where thcrc is parakeratosis, and it is absent. where there is hypcrkeratosis in papillary hyperplasia of the palate. 2. Acanthosis and inflammation arc seen in areas where there is alycogen deposition. Ko relationship is infrrretl from this stntly. 3. Glycogen is seen in the prickle-cell layer, in “fissures,” aronnd epithelial pearls, and, rarely. in thr basal cells of pa.pillary hypcrplasia of the palate. 4. In sonic instances, v;\.cuola.tctl c~l)ithclial ~11s caontaill heavy tlrposits of glycogen. REFERENCES 1. Smith, W., and Gault, S.: Essentials of Pathology, New York, 1942, D. ~~l’l)l(:toll-Cc!lturv Company, pp. 17-18. Lesions of the Oral Regions, D. 2. Robinson, H. B. G.: Neoplasms am1 “Precancerous” Clin. North America, pp. 621-626, November, 1957. Papillary Hpperplasla, J. Oral Surg., Anesth. 8r Hosp. T). 3. Waite, I).: Inflammatory Serv. 19: 210-214, 1961. 4. McManus, J. F. A., and Mowry, W.: Staining Methods. Histologic anll Histochemical, New York, 1960, Paul B. Hoeber, Inc., pp. 63-64. ljistribution of 5. Klingsherg, J., CaneelIaro, A., and Butcher, 0.: The Comparative and Acid Mucopolynaccharitles ill ljifferentGlycoge?, Succinic Dehydrogenase, aged Animals, J. D. Res. 40: 461-469, 1961. 6. Cahn, L., Eisenbud, L., and Blake, N.: Histochemieal Analysis of White Lesions of the Mouth. II. Analysis of Glpcogen Content, ORAL F~JRG., ORAL MED. k ORAL PATH.
15:
458-462,
1962.
7. Turesky, S., Glickman, I., and Provost, Process in Oral Lesions Diagnosed MED. & ORAL-PATH.
14: 442-453,
.J.: A Histochemieal Study of the Keratotic Clinically as Leukoplakia, ORAI. STW:., ORAT.
1961.
Stu(ly of the Attached 8. Turesky, S., Fisher, B., and Glickman, I.: A Histochemical Gingiva in Pregnancy, J. D. Res. 37: 1115-1122, 195s. !!. I)ewar, R.: Observations on the Composition and Metabolism of Normal ant1 Tnflametl Gingivae, J. Periodont. 26: 29-39, 1955. 10. Turesky, S., Glickman, I., and Litwin, R.: A Histochemical Evaluation of Normal and Inflamed Human Gingivae, J. D. Res. 30: 792-798, 1951. White Folded 1)ysplaaia of the I I. Zegarelli, E., Everett, F., and Kutscher, A.: Familial l\lucous Membranes, Oaar, SURG.,ORAr,M&D. & ORAL PATH. 14: 1436.1443, 1!161,