of roots adjacent to chronic periodontal pockets, may
make optimum ranges determined in this animal study
slighth low. Tor human pocket reutluchment techniques,

The assistance of surgical technicians. James Kent. Robert
Murraj and Walter Paschal I and histolog) technicians. Neva
Svkes. Bern Crismun and Leslie Cupp, is gralefull) acknowledged.
K I I I R I N( I S
I- Register. A. A.: Bone and cement um induction b\
dent in, deminerali/ed in situ.J Periodontol 44: 49. 1973.
2. Register, A. A. el al. Human bone induction by
aliogeneic dentin matrix J Periodontol 43: 4>9, 1972.
3. Urist, M. R.: Bine hislogenesis and morphogenesis in
implants ordemineralized enamel and dentin. OralSurg 29: 88,
1971
Accelerated Reuttachmem 655
4. Yeomans, J. IX. and Urist, M. R.: Bone induction bv
decalcified dentine implanted into oral, osseous and muscle
tissues. Arch Oral Biol 12: 999. 1967.
S Bang, Ciisle. and Urist. M. R.; Bone induction in
excavation chambers in matrix of decalcified dentin. Arch Surg
*>4: 7X1, 1967.
6. Duhue. F. 1... and Urist, M, R : The accessibtlil) of the
bone induction principle in surface-decalcified bone implants.
Ctin On hop 55: 217. 1967.
7. Urist. M. R.: Surface-Decalcified Allogeneic Bone
(SDAB) Implants Ciin Orthop 56: 37. 1968,
8 Younger. W. J.: Lactic acid in pvorrhea. Am J Dem Sci
31: 334. 1X97 98.
9. Younger. W. J.: Pyorrhea alveolaris, schweizerische
viertelpahrschrift fuer /ahnheilkunde. Zuerich 15: 87. 1905.
10. Slew art. H. T.: Partial removal of cement um and
decalcification of tooth in the treatment of pvorrhea alveolaris.
Dent Cosmos 41: 617. 1899

Abstracts

Mi I v s i \ s i v r o m i
Perlmulter, S.. Buchncr. V. and Smukler. H
Oral Surg J8: 749, November. 1974.
An unmarried Caucasian woman, age 44 had .i radical mastectomy
performed for removal ol a tumor followed bv postoperative radiotherapy.
About eight months later, -.he developed generalized redness and
swelling of her gingiva and was referred to a periodontist. The upper
nghi premolar area revealed, upon examination, a pedunculaled mass
which extended OUT two maxillary premolars. Clinicall) it resembled
either a peripheral giant tell granuloma or a pyogenic grunulomu. An
excisional biopsv was performed and hislopalhological diagnosis
confirmed it as an anaplaslic carcinoma with a good chance lhal il was
a meUslatic lesion, hew casts of metastasis to the gingiva without bone
involvement have been reported- Faculty ol Continuing Medical
Education. Dental Division. Tel 4viv University, Tel Aviv. Israel
I M M U N O F L U O R E S C E N T D E T E R M I N A T I O N 0 1 I Ml R n v l U I
PROPORTIONS I>I Streptococcus Muwns i\ Hi v i \ s D E N T A L fJi.Ayi v:
A COMPARISON W I I U CULTURAL TECHNIQUES
Kmilson. C. (j., Kohler. H.. and Bratlhal
Arch Oral Biol 20: 81, Januarv, 1975.
I)
The relative1 proportions o(Streptococcus mulans in sonified plaque
smears were determined wilh the immunolluoreseence technique and
compared with the plaque samples which were cultivated on blood agar
plates, mitissalivarius. (MS], and Ml' agar plates. The percentage of
Streptoccitcus mulans in 36 samples ^.is 7.3 • »ilh fluoreseein-isothiocyanate
labeled antisera and 6.1% b> cultural methods. The determination
with the immunolluoreseence technique showed a linear proportion
with the other methods. A few cases had a higher value in
fluorescence when compared vsiih MC agar. Department of Cariology,
Faculty of Odontology, University of G'oleborg, S-4CM) 33 Goteborg.
Sweden
FORMATION ot SUPTORTINC BONE I\ ASSOCIATION WITH
PERIODONTAL LIGAMEM ORGANIZATION IN THE MOUSE
Ten Cate, A. K.
Arch Oral Biol 20; 137, February, 1975.
In a previous siudv b> implanting tooth germs subcutaneously it was
shown that the ectomesench) mal cells surrounding the dental organ and
continuous wilh the dental papilla most probahh give rise to the
cemenlum, periodontal ligament, and alveolar bone. This study to
determine whether ihe tooth actually produced its own "alveolar" bone
involved injecting 5 jimc'g b,w. of tritriated proline into mice at various
intervals. The mice were then sacrificed on different occasions, lower
laws dissected out. and examined microscopically. It was shown thai
bone deposition occurred after 12 or 13 days and was coincident with
periodontal ligament formation. However, il was not possible to
determine where the bone originated, although it was suggested that it
may originate from a layer of ectomesenchymul cells investing the tooth
germ. Fatuity of Dentistry. University of Toronto, Toronto. Canada
t u r n OH MONIHIV PROPHYLAXIS ON CARIES VMI GINGIVITIS IN
SCHOOI f HILDREN
Badersten, A., Egelberg, J-, and Koch, G.
Community Dent Oral hpidemoil i: I, Februarv. 1975.
Prophylactic treatment with monthlv intervals were given to 216
schoolchildren, divided into experimental and control groups. The
prophylactic treatment included instruction and practice in toothbrushing,
dental floss, and toothpicks for interpro\imal surfaces, and 0.2^
sodium fluoride rinses under supervision of a dental nurse. Thev spent
15 minutes for each session. There was no differences in reduction of
caries obtained for occlusal, bltccal and lingual surfaces, bui significant
reductions were observed for the proximal surfaces. Gingivitis was
reduced in the experimental group during the year of trial The effect of
treatment was more noticeable in the group with high caries experience.
Prophylaxis every second week appeared to be more efficient than every
month. School of Dentistry, Department of Periodoniology. University
of Lund, S-214 21 Malmo, Sweden

Osseous Autografts

II. Histologieal Responses to Osseous
Coagulum-Bone Blend Grafts
by
S. J. FROUM
R. THALER
I. W. SCOPP
S. S. STAHL

THE CURRENT INVESTIGATION is a continuation of a
previously reported study which dealt with the responses
of intraosseous defects to a variety of therapeutic
procedures.1 In the initial report we reviewed clinical and
histological literature pertinent to the subject and also
described clinical responses to the placement of either
osseous cougulum-bone blend or autogenous iliac marrow
and canccllous bone grafts into human periodontal
defects. Our results concurred with those of Hiatl and
Serial Ihorn1 which suggested that similar levels of osseous
regeneration apparently took place regardless of
graft material used.
In order to further stud\ the behavior of osseous
coagulum-bone blend grafts in human infrabony defects,
three cases will be presented in which histologic findings
of block sections were obtained from the graft site 6 to 13
weeks postsurger>.

MATERIALS AND METHODS

All patients in this investigation were referred for
periodontal therapy to the Periodontal Section of the
Dental Service at the Veterans Administration Hospital
in New York City. Although they were in excellent
health at the time of the study, each suffered from severe
periodontitis. Three patients were selected with infrabony
defects on anterior teeth scheduled for extraction. These
teeth were adjacent to teeth which were to be retained,
but which also had infrabony defects requiring osseous
surgery. Each patient signed an informed consent form.
Initial periodontal therapy was similar in all patients.
It consisted of oral hygiene instruction, scaling, occlusal
adjustment, root planing and cureltage under local
anesthesia. An oral hygiene index was utilized, and only
when the index approached zero was surgery performed.
During initial therapy two sets of study models were
constructed. The presurgical and poslsurgical measurements
of pocket depths and osseous recontouring were
described in detail in our previous publication.1.
Periodontics Section. Dental Service, New York Veterans Administration
Hospital and the Department oT Periodontics, New York
University College ol Dentistry at the Brookdale Dental Center, New
York. N. Y.
Prior to block section, radiographs were taken and soft
tissue measurements were recorded. At this time, osseous
measurements were made through intact tissue using the
stent as a fixed reference point. The accuracy of the latter
technique has been described.'
Block sections were removed following the technique
described by Dragoo and Sullivan." Healing and prosthetic
replacement of the extracted teeth occurred without
complication in all three cases. The clinical aspects of
the three cases will now be described in detail.
Case 1
Pertinent Medical History. A 64-year-old male presented
for periodontal therapy. The patient's medical
history revealed that the patient suffered from a hcrniated
disc during the Second World War. A laminectomv
was performed soon afterward. The patient also
revealed a history of hepatitis in I960. He had a lipoma
removed in 1972 but was currently in good health.
Pertinent Periodontal History. The mandibular central
incisors and left lateral incisor had such severe periodonlal
involvement that these teeth were recommended for
extraction. The left canine (which was to be retained) had
an infrabony defect on its mesial aspect. To reduce the
mobility of the left central incisor (Class II). an intracoronal
wire was used to fabricate a splint between the
central and left lateral incisors.
Periodontal measurements on ihe left central incisor
(tooth #24) were recorded prior to and during osseous
coagulum-bone blend implant surgery. The initial pocket
depth on the mesial of this tooth was 7.2 mm. An inverse
bevel full thickness tlap was reflected revealing a two wall
infrabony defect with an osseous depth which measured
5.9 mm using the stent as a fixed reference point (Kig. I).
The defect was debrided, root accretions were removed,
and the defect was irrigated and overfilled with osseous
coagulum-bone blend. The flap was then coapted and
sutured to insure maximum soft tissue coverage. A
periodonlal dressing was applied and the patient was put
on antibiotic coverage for one week. Thirteen weeks
posloperatively. immediately prior to block section,
measurements were again recorded. Pocket depth now
measured 3.9 mm. Clinical crestal bone sounding, utilizing
the stent, revealed an apparent osseous fill of 3.4 mm.
The healing after block section was uneventful.
Case I!
Pertinent Medical History. A 40-year-old male presented
for periodontal therapy who had been treated at
the NYVAH one year earlier for a bilateral mandibular
fracture. He also revealed a history of syphilis in 1970.
The patient was currently in excellent health.
Pertinent Periodontal History. In order to replace the
maxillary left lateral incisor and because of the severe
overbite and flaring of the remaining incisors, these teeth
were scheduled for extraction. The distal aspect of one of
these teeth (tooth §1) as well as the mesial of tooth #11.
656
Volume 46
Number 1!
which was to be retained, had infrabon> defects. Periodontal
measurements on tooth #7 were recorded prior to
and during osseous coagulum-bone blend implant surgery.
FIGURE i, Clinical appearance ot osseous crater at time of
initial surgery. Probe placed into osseous defect. Case I.
Osseous Auiografis, II 657
The preoperative pocket depth of the distal of tooth
§1 and 8.1 mm. An inverse bevel full thickness Hap was
reflected revealing a one wall hemiseptum with an
osseous depth which measured 4.3 mm using the stent
(Figs. 2A and 2B). The delect was debrided, root
accretions were removed, the defect was irrigated and
overfilled with osseous coagulum-bone blend. The Hap
was then coapted and sutured to insure maximum soft
tissue coverage. A periodontal dressing was applied and
the patient was put on antibiotic coverage for one week.
Six weeks postoperatively. immediately prior to block
section, measurements were again recorded. Pocket
depth now measured 4.0 mm. Clinical crestal sounding,
using the stent, revealed an apparent osseous fill of 2.6
mm. The healing after block section was uneventful.
Case III
Pertinent Medical History. A 53-year-old male revealed
a history of treatment for depression in 1971.
Presently, the patient was in excellent physical and
mental health.
FIGURE 2. A. Clinical appearance of osseous crater at time ot initial surgerv. Case II: B. Probe in position at osseous crater ot Case
II at lime of initial surgery.

FIGURE 3. A. Overview o/ a mesio-dista! cut section of the aratt site. Case I. mesial ot tooth #24. Tooth and tissue removed 13 weeks
after graft had been placed. Hematoxylineosin stain. Magnification 10 •. B. Higher magnification of arrowed area shown in Figure
3A . Sote new cementum formation at xrai> site and osteogenesis al alveolar seams. Magnification 25 * . C. Further magnification ot
lien cementum seen in Figure IB. Sote the relative lack ofcellularii v in the cementum and the presence of lamellae. Magnification 64
x.
658 Froum, Thaler, Scopp, Siahl November, I9"?5
D
[•K,r KI 4. A, Parallel orientation i// periodontal ligament libers between bone and cemenlum seen in Fig, JA. Also note osseous
remodelling ai the wall. Magnification M • B. New bone formation at alveolar wall {arrow) from section shown in Figure 3A. C.
New cementum forming on deniin from section shown in Figure 3 1. Magnification h4 <. D. Osseous remodelling at crater wall from
section shown in Figure 3A. Magnification f>4 • ,
Pertinent Periodontal History. The mandibular centrals
and left lateral incisor v,crc diagnosed for extraction.
The distal of the right central incisor (looth #25)
and mesial of the left canine (tooth #22), which was to be
retained, had infrabony defects. Prior to initial therapy
an intracoronal splint was conslructed on teeth #23 to 25
because of the severe mobility of looth §25. Prior to and
during osseous implant surgery, periodontal measurements
were recorded on toolh #25. The preoperative
pocket depth on the distal of this tooth was 8.0 mm. An
inverse bevel full thickness flap was reflected. Root
accretions were removed and the defect was debrided. It
was classified as a combination one wall hemiseptum
with two walls at its base. The osseous depth of the defect
measured 3.4 mm using the stent. The defect was
overfilled with osseous coagulum-bone blend. The flap
was coapted and sutured to insure maximum soft tissue
coverage. A periodontal dressing was applied and the
patient was put on antibiotic coverage for one week.
Nine weeks postoperatively, immediately prior to
block section, measurements were again recorded.
Pocket depth was now 2.2 mm. Clinical creslat sounding
revealed an apparent osseous fill of 2.4 mm. Healing
after block section was uneventful.

HISTOLOGIC OBSERVATIONS

At the outset, it must be stated that histologic evidence
based on three specimens only indicates possible rather
than probable histologic responses. And it is within these
limitations that the repair phenomena observed in our
specimens must be evaluated. For clarity, responses will
be divided by anatomical sites, i.e., histologic activities al
FlCl Kl 5. Spicules bfittg exfoliated in specimen removed 6
weeks after graft had been placed. Case II. distal of tooth #7.
Mesio-disial section. Hematoxvlin-eosin slain. Magnification
10 x.
Osseous Autografts, II 659
the (I) ulveolar walls. (2) bonv implants. (3) periodontal
ligaments, and (4) cementum. Since all three specimens
showed similar responses, overall histologic trends will be
reported.
Alveolar Walls. In our specimens, osteogenesis was the
most frequent response seen at the osseous seams. In
other areas, osteoblasts lined the seams, but osteoid was
not visible. Although evidence of resorption-apposition
remodeling was present along all walls, our material
obviously could not indicate whether or not this remodeling
occurred pre- or postsurgical therapy. However,
osteoclastic activity and appearance or resorplion bays
were relatively limited (Fig. 3A 3C\ 4A 4D).
Bony Implants. Bony implants were seen in all specimens.
In some sections, spicules were being exfoliated
(Fig. 5). Yet, despite this exfoliation, spicules within the
defects showed evidence of osleogenesis and osteoelasis.
But again, the predominant impression was that most
spicules observed within the delects were either lined bj
osleoblasts or demonstrated both osteoid and an osteoblast
lining at their borders. Resorption appeared limited.
Some of the spicules contained osleocytes within
their lacunar spaces, while others appeared empty. The
latter observation may reflect loss of vitality of the
spicule, or be due to the plane of section. In other
spicules, osteocytes were present in some portions of the
specimen, but not in others (Figs, 6A 6C). Furthermore,
it seemed that larger spicules were more apt to contain
empty lacunar spaces and demonstrate peripheral resorption
than smaller spicules.
Periodoniat Ligament. The periodontal ligament appeared
exceedingly cellular. Connective tissue lined the
osseous walls and the spicules. In some areas, the
periodontal ligament was functionally oriented with
fibers bundles apparently inserting al either the osseous
walls or the borders of the spicules al one end and al the
h l<ji RI 6. A. Spicules in area of graft from specimen shown in Figure 5. Sole evidence ofosleogenesis (arrow) and osteoelasis at the
borders of the spicules. Magnification 64 x . B. Spicule in area of graft from specimen shown in Figure 5. \'ote active connective tissue
surrounding spicule. some osteoblasts lining spicule and variations in lacunar content. Magnification 64 -:. C. Spicule demonstrating
empiv lacunae and osteogenic activity in graft siie. Case I. Magnification 64 x.
660 Froum, Thaler, Scopp, Siahl
J. Perioduniul.
November. 1975
newly formed cementum at Lhe opposing root surface
(Fig. 7). In other areas, the orientations of the ligament
appeared to run parallel to the root surface (Fig. 4\\.
Root Surface. The root surfaces facing the implants
1
FiCii KI 7. Functionally oriented periodonial fibers between
spicule and new cementum [arrow) Irani specimen shown in
Figure 5. Magnification f>4 x .
showed evidence of cemental resorption extending into
dentin in some areas, but nol in others. Again, our
present material does not allow for conjecture as to the
time when these rcsorptive processes occurred, However,
il was striking lo nole significant amounts of "new"
cementum which was present at the root surfaces in the
area of implants. This cementum was essentially acellular,
slightly tamellated and had formed on old cementum
as well as on dentin (Figs. 3C and 8A). Furthermore,
some cemental apposition was also present at root
surfaces of teeth immediately adjacent lo the teeth
surrounded by the osseous defect. However, this cemental
apposition was significantly less than thai seen
at the tooth wall facing the implant (Fig. KB). Cementoblasts
appeared to be lining the new cementum
and collagen insertion or parallel collagen fill was noted
along these root surfaces.
COMMEN I
To the best of our knowledge, this is the first histologic
evidence of the response of the penodontium to the
"bone blend" type of osseous coagulum. In general, the
periodontal response appeared similar lo what has been
described with other autogenous bone grafts in previously
published observations.*- *• ' " 7
For example, osleogenesis has been reported in experimental
and human hisiologic material as early as two
weeks after surgery5-fi and was found to be significantly
active for at least two months after implantation.4
Cemenlogenesis has been reported to take place wilhin a
similar time span.4- s
Smaller si/e bone particles have been described to
FKH KI 8. A. New cementum formed on old cementum in area of graft from specimen shown in Figure 5. Magnification 64 x B.
New cementum forming on mesial mot of tooth #6. Case II. This tooth was adjacent to tooth #7 where the ma/or osseous delect was
located. Note the lesser amount of new cementum. Magnification 64 x.
Volume 46
Number II
induce more bone formation than larger pieces* which
concurs with our observations. Along this line, it should
be remembered that functional remodeling probably
continues at an implant site, albeit at a slower pace, for
many months or years after surgery as a response to
continuous function,!- ?- * but the most active phase appears
to take place within two to three months after
placement of the graft.
A point of special interest was the variations in
orientation of periodontal ligament fibers which were
seen both within the same specimen as well as between
specimens. These observations must be coupled with the
report by Kllegaard and associates6 who in experimental
studies of graft healing stated: "even after 24 weeks very
few areas showed well defined bundles of collagenous
fibers attached to the new cementum. Usually, the fibers
of the periodontal ligament were oriented parallel to the
root surface. However, fibers oriented parallel to the root
surface could also be seen further apically where no
surgery had been done." On the other hand, in an
eight-month postsurgery human specimen, Dragoo and
Sullivan" reported a "fairly mature, functionally oriented
periodontal ligament." Our observations support the
thesis that fibers early in the repair process orient either
functionally or parallel to the root. The reason for this
variation in orientation is not known. Since histology
obviously cannot trace a continuous process in the same
specimen, it relies on sequential specimens for disclosure
of a continuous process. At this time, human graft
specimens are limited in number. Thus, it may well be
true that initially parallel oriented fibers adapt a more
functional orientation with time. On the other hand,
parallel orientation may occur in the early phases of
healing as a result of as yet unknown factors in the
repairing milieu and remain in that orientation. Further
study of this phenomenon is obviously needed, since it
may be of clinical significance.
Finally, our present findings demonstrate an apparent
enhancement of cementogenesis at the site of implant
similar to that reported by Dragoo and Sullivan.* Since
We did not use iliac bone graft, it appears that not only
"undifferenliated cells" may enhance cementogenesis,
but also tissues transplanted in a "bone blend."
Osseous A utografts, II 661
SUMMARY
Three human intraosseous lesions were treated using
osseous coagulum-bone blend as graft material. These
sites were surgically removed 6 to 13 weeks after
treatment and the blocks prepared for histologic analysis.
Periodontal remodeling at the site of grafting was
noted in all specimens. This remodeling involved the
osseous walls, periodontal ligament, ccmentum and graft
spicules. Both osteoclastic and osteoblastic activities
were seen at the borders of the spicules. Exfoliation of
spicules was particularly frequent in the six-week specimen.
Nevertheless, regeneration of both bone and cementum
was actively taking place even in ihis early specimen
and was still evident in the older specimens.
Of particular interest was the apparent marked increase
in cementogenesis at the graft sites and the
variations in parallel or functional orientation of the
periodontal ligament in these areas. Significant functional
orientation of segments of the periodontal ligament
were seen in the early specimens, but not in the
latter one. The reason for such variations in ligament
pattern is not known at this time.
REFEREM IS
1. Froum, S. .1.. Thaler. R.. Scopp. I. Vs.. and Stahl. S. S.:
Osseous Autografts I. Clinical responses So hone blend or hip
marrow• grafts. J Periodontol46:5!5, 1975.
2. Hiatt, W. H.. and Schullhorn. R G.: Immoral transplants
of cancellous bone and marrow in periodonuil lesions. J
Periodomoi 44: 194, 1973.
3. Schallhorn. R. G.. Hiatt. VV. H., and Boyce, W.: Iliac
transplants in periodontal therapy. J Periodonlol 41: 566. 1970.
4. Dragoo. M. R.. and Sullivan. H. C: A clinical and
histological evaluation of autogenous iliac bone grafts in
humans: Part 1. Wound healing 2 to K months. J Periodomoi
44: 599. 197.1.
5. Rivaul!, A. F., Toio, P. 1).. Levy, S.. and Gargiulo, A.
W.: Autogenous bone grafts: Osseous coagulum and osseous
retrograde procedures in primates. J Periodomoi 42: 7X7. 1971.
6. Ellegaard, B.. Karring. T., Lislgarten, M.. and Loe. H.:
New attachment after treatment of inlerradicubr lesions. J
Periodomoi 44: 209, 1973.
7. Ross, S. ti.. and Cohen. D. W.: The late of an osseous
tissue autograft. Periodontics 6: 145. 1968.
X. Nabers. C. L.. Reetl. O. M.. and Hamner, III. J. E.:
Gross and histologic evaluation of an autogenous none grail 57
months posioperatively. J Periodomoi 43: 702, 1972.

The Effect of Endotoxins and
Enzymes in Vitro on the Release
of Gingival Histamine*

by
JOSHFH SCHWARTZ, D.D.S., vi.s.n.t
MARTHA DIBBL.LL, B.A.J
BACTIRIAI PLAQUE and their products have been implicated
as the primary eliological factors contributing to
inflammatory periodonlal disease.1 2 One of these bacterial
products, the endotoxin, has recently received much
attention in the literature. Triliated endotoxin was observed
to penetrate intact creviLular epithelium in dogs.3
Upon entry into the connective tissue in rabbits, endotoxin
was shown to elicit a Shwartzman reaction,4 and to
effect a local antibody production in regional lymph
nodes/1 The quantity of endotoxin in gingival exudate has
been correlated with the degree of gingival inflammation.*
Enzymes of bacterial origin also have been considered
etiological agents.7 However, in periodonlal disease,
enzymes can originate from tissue cells as well as from
bacteria. The enzyme hyaluronidase has been shown
histologically to increase the intercellular space.8 It acts
to break down mucopolysaccharides of the intercellular
substance of the epithelium and the ground substance of
the gingival corium. This enzyme may allow further
penetration of other bacterial products. Collagenase, an
enzyme produced by Bacteriodes melaninogenicus, an
inhabitant of the crevice,9 and by gingiva in vitro,"1 has
the ability to destroy and lyse native collagen. Similarly
proteases, enzymes produced by bacteria, cell infiltrates
(PMN's) and tissue cells, can also result in the destruction
and lysis of connective tissue."
The exact role of endotoxins and enzymes in periodontal
disease is, as yet, unclear. However, it is possible that
these agents may penetrate the crevicular epithelium,
enlarge the intercellular space, cause vasodilation and
vascular leakage, initiate chemot.jctic activity and lysosomal
enzyme discharge, influence tissue cells to release
chemical mediators and enzymes, destroy native collagen
and ground substance, and promote immediate and
delayed hypersensitivity reactions. But, evidence for their
participation in the generation of periodonlal disease is
lacking.
Shelton and Hall" have recently reported that healthy
gingiva is rich in mast cells and that these cells become
reduced in observable numbers during inflammation.
Mast cells contain histamine" which upon release can
cause capillary permeability and vasodilation. These two
vascular changes are early events in inflammation."
Does the passage of endotoxins and enzymes through the
gingival corium release vasoactive mediators from the
gingiva.'
The purpose of this investigation was to determine if
endotoxins and enzymes, which have been implicated as
etiological factors in periodontal disease, effect gingival
histamine release.
METHODS AND MATERIALS
The buccal gingival tissues of fifteen Beagle dogs were
used for this study. Their teeth were scaled, two weeks
prior to experimentation and brushed daily thereafter.
The dogs wercanesthesi/ed with sodium penlabarhital 50
mgs/ml. During the procedure the dogs were maintained
at stage Ill-plane III of anesthesia. A band of attached
gingiva was removed from one quadrant and was immediately
placed in Hank's Balanced Salt Solution. The
gingival sample was sectioned into small pieces I x l x l
mm, part of which served as the uncultured specimen: the
remaining pieces were placed in organ culture dishes*
containing the various test substances. The cultures were
gassed with a mixture of 5% CO2, 50% O,. 45% N2 for 10
minutes and incubated at 37°C for four hours. The
uncultured pieces of gingiva were drained, weighed and
Iro/en in liquid nitrogen to serve as an uncultured
control. The test substances used were Hank's Minimal
lissential Medium (MEM)t (culture control medium),
compound 48/80,1 endotoxins,§ eollagenuse,j| hyaluronidase,!
1 protease,** trypsin.tt and chymotrypsin.it
Each medium contained MEM as a diluent and was
prepared fresh prior to incubation. At the end of the
incubation period, the media and tissues were recovered.
The media were placed in small scrum bottles and frozen
in liquid nitrogen. Tissue pieces were drained, weighed
and frozen as above. The mean weights of the tissue
are shown in Table I.
The histamine content of the uncultured tissue, cul-
* This research vv;is supported b) (he l.nited Slates Public Health
Service from N.I.D.R. (ir;int DE 03265-01-03.
t Department of Permdoniulogv, University ul Oregon DenUil
School, Portland, Oregon 97201.
] University of Oregon Dental School, Portland, Oregon 47201.
* I alcon Plashes, Oxnard, California
t I''! Glutumine. 100 Li/ml antibiotics and bicarbonate to pH 7.4
were added to the medium—Grand Island Biological Co., Grand
Island. Ne» York
I Compound 48/K0 0.! to 100 tig/ml. Burroughs, Wellcome and
Company, Tuckuhoe. New York
§ Endotoxins, 100 jig/ml, Difco Laboratories, Detroit, Michigan;
Escherichia coh Endotoxin OI27:B8; Salmonella lyphosa lindotoxin,
0901; I.epiolnchia butx'ulis Endotoxin (phenol soluble) courtesy of Dr.
Kenneth knooi.
I1 Collagenase, 30 N.F. units/ml, cl. histolyticum Type t, Sigma
Chemical Co.. St. Louis. Mo.
T Hyaluronidase, 50 N.F. units/ml, bovine testes Type I. Sigma
Chemical Co.
** Protease Type VII, 0001 to KM) prololytie unils/ml. Bacterial
(pfs), Sigma Chemical Co.
t t T r y p sm Type III. 10 to 100,000 BAEE Units/ml, pancreas 2 •
crystalized, Sigma Chemical Co.
tt Chymotrypsin Type IV, 10 to 10.000 chymotrypsin unils/ml
BTtE Units, from bovine pancreas, 3 x crystalized, Sigma Chemical
Co.
/
662