Effect of X-irradiation and Beta Emanation on Circulation in the Hamster Cheek Pouch
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Effect of X-irradiation and Beta Emanation
on Circulation in the Hamster
Cheek Pouch
By GEORGE P. FULTON, P H . D . , BRENTON R. LUTZ, P H . D . AND ROMA KAGAN, A.B.
The small venules of the hamster cheek pouch were rendered susceptible to hemorrhage by total
body x-irradiation, cheek pouch x-irradiation and beta emanations from strontium-90 glass beads
imbedded in the cheek pouch, as determined by negative pressure and moccasin venom tests. Pe-
techiae were formed without vessel rupture and ceased without formation of platelet plug or fibrin
clot. Increased susceptibility to petechiae preceded thrombocytopenia. Endothelial damage was
indicated by increased leukocytic adhesiveness to walls of venules. Vasodilation, tortuosity, newly-
formed capillaries, and occasional "sausage-shaped" segments of constriction in arterioles were
characteristic vascular responses. Near death, erythrocytes circulated in aggregates varying from
typical rouleau formation to discrete masses, resembling "sludged blood" but without formation
of occluding thrombi.
S INCE the discovery of extensive bleeding
into the tissues in victims of the atom
bomb at Hiroshima, the importance of
hemorrhagic defects resulting from ionizing ra-
Lushbaugh and Hale 1 described the vascular
reactions within rabbit ear chambers subse-
quent to total body x-irradiation with 1500 r.
Hyperemia, leukocytic adhesiveness to vessel
diation has been recognized. Cronkite and co- walls and circulating erythrocytic aggregates
workers1 have discussed possible causative fac- were reported.
tors and mechanisms which may be involved, This paper describes the hemorrhagic effects
but results obtained from the method of direct of irradiation of the hamster and the changes in
capillary microscopy were not available for peripheral circulation as shown by direct mi-
analysis at that time. Although thrombocyto- croscopic observations and motion picture re-
penia was implicated as one of the most impor- cording of the blood vessels and flow in the
tant factors in the bleeding defect,1 other possi- everted cheek pouch. Several types of irradia-
bilities were not entirely eliminated, such as a tion were used for comparative purposes: first,
defective vessel wall or perivascular sheath. exposure of the total body to single doses of
Although direct microscopy of the minute x-ray; second, whole or partial cheek pouch
blood vessels might conceivably contribute to x-irradiation; and third, chronic exposure to
our understanding of the vascular effects of imbedded radioactive fragments 03-emitting
ionizing radiation, relatively few extensive in- strontium glass beads and 7-ray cobalt needles).
vestigations of this type have been reported.
Smith, Svihla and Patt 2 described the changes METHODS
induced in the wing of the bat by whole and A well-shielded, compact x-ray unit was custom-
partial body x-irradiation. However, the bat built* for small animal work, consisting of a
Machlett x-ray tube, water-cooled and housed in
was found to be extremely resistant, since the top of a metal cabinet (21" x 21" x 4S") con-
15,000 to 60,000 r were required to shorten the taining adjustable shelves and lined with lead 1^"
life span. Furthermore, hemorrhages were ab- in thickness. The cabinet is mounted on a steel
sent even with the highest doses used. Moseley, bench with working space for microscopic observa-
tions or other purposes. The transformer is located
From the Department of Biology, Boston Uni- beneath the bench and mounted on rollers in order
versity, Boston, Mass. to facilitate servicing. The x-ray cabinet, bench and
This investigation was supported by Atomic En-
ergy Commission Contract No. (30-l)-897. * Campbell X-ray Corporation, 110 Cummington
Received for publication October 10, 1955. Street, Boston, Massachusetts.
133 Circulation Research, Volume 1 V, March IMS
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FIG. 1. A, Hamster cheek poucli everted and spread for transillumiiiation of vascular bed. B, Lead-
lined .shield with removable cover and perforated "stove-lids" for spot x-irradiation of the everted
cheek pouch. C, Leukocytic adhesiveness to venular endothelium at 4 to 6 hours after total body
x-irradiation with 1200 r. Original magnification 220 X. Vasodilation and increased vascularity in
hamster cheek pouch (E) at 24 hours after total body x-irradiation with 1200 r, compared with appear-
ance before irradiation (D). Original magnification 7 X.
transformer are mounted on a cement slab and the measured in air at the surface by means of a Vic-
control panel rests on the floor nearby. Maximal toreen r-meter.
values for the irradiation characteristics obtainable At various intervals following irradiation, the
with this unit are 150 kv, S ma, and 125 r/minute cheek pouch was everted (fig. IA) in hamsters
at a 10-inch target distance. anesthetized with Nembutal (10 mg./lOO Gm. body
Golden hamsters (Mesocricetus auralm)* were weight). The characteristics of blood flow and the
given total body x-irradiation without anesthesia, conditions of the intravascular elements were ob-
while confined individually in thin plastic tubes. served, and significant variations were recorded
Several hundred young adults (10 to 12 weeks) of cinephotomicrographically according to the method
both sexes were used for the various procedures in described previously.4
this investigation. The control panel was regulated Entire or partial cheek pouch x-irradiation was
to provide irradiation factors of 145 kv., 7 ma and accomplished in anesthetized hamsters with the
100 r/minute at 10 inches. Xo filtration was used cheek pouch everted and prepared for transillumi-
except for that inherent in the tube. The dose was iiation prior to irradiation. The transilluminating
dish with optical block was mounted in a custom-
built metallic container with a removable lead-lined
* Procured from Golden Nugget Hamstery, Mav- cover, perforated for air exchange (fig. IB) A
nanl, Muss. "stove-lid" arrangement in the portion of the cover
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Fici. 2. A, Rouleaux formation in blood vessel of hamster cheek pouch during erythrocytopenia
induced by total body x-irradiation. Original magnification 520 X. H, Circulating erythroeytie
aggregates prior to death of hamster receiving lethal x-irradiation. Taken from 10 mm. Kodachromo
motion picture soquence. Original magnification 520 X. Viisodilation (D) produced by spot x-irra-
diation (e.g., 50 r delivered to circular area 10 mm. in diameter), compared with same cheek pouch
before irradiation (C). Inert glass bead (diameter 0.5 mm.) used for identification of exposed area.
Original magnification 7 X. E, Tortuosities of arterioles, capillaries and venulcs at 24 hours after
x-irradiation of cheek pouch with 15,000 r. Minute petechial hemorrhages are shown at venular
junctions in vicinity of inert glass bead marker. Original magnification 7 X. h\ "sausage-shaped"
portions of blood vessels characterized by localized constrictions and dilations. Original magnifica-
tion 7 X.
above the cheek pouch permitted x-inadiation of peripheral incision in the epithelium and massaged
small circular areas of the pouch through holes to a central position remote from the incision
with diameters of 0.5, 1.5, 5, 10 and 25 mm. By the (fig. 2C and D). The bead was imbedded carefully
elimination of all "stove-lids", the entire cheek at least seven clays before irradiation and retained
pouch could be irradiated, with the I'est of the the original position without irritation. The portion
hamster shielded. By using a non-perforated "stove- of the cheek pouch containing the bead was centered
lid" and by removing the major portion of the under the opening in the "stove-lid" during irra-
cover of the container, the everted cheek pouch diation. Periodic post-irradiation studies of the
could be shielded while the rest of the hamster irradiated spot were possible because the glass
was irradiated. In order to mark the spot selected bead marker assured accurate and ready identifica-
for irradiation, a non-irritating glass betid (approxi- tion of the exposed area.
mately 0.5 mm in diameter) was inserted in the Chronic beta irradiation was accomplished by
connective tissue of the pouch through a small implanting radioactive teids, approximately 1 mm.
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FIG. 3. Increased vascularity and petechial formation (5) in vicinity of radioactive strontium
glass bead (activity 140 NC) imbedded in subepithelial tissue of cheek pouch for two days, compared
with same pouch at time of implantation of bead (A). Original magnification 7 X. Tortuosity, in-
creased vascularity and petechial formation (Z)) in vicinity of cobalt needle (activity 92/ic) imbedded
in subepithelial tissue of cheek pouch for 6 weeks, compared with same pouch at time of implantation
of needle (C). Original magnification 7 X.
in diameter (fig. 3A and B) and composed of glass to divert the bloodflowinto an alternate vessel.
fused with strontium 90,6* into the subepithelial Some of the adhering leukocytes appeared to
connective tissue of the cheek pouch through an
incision in the mucous membrane. The radioactive
lack the property of ameboid movement, per-
beads were massaged into a position remote from haps due to injury as a result of the irradiation.
the incision. The cheek pouch containing the im- The peripheral leukocytosis persisted for 8 to 12
bedded bead was reinserted in the buccal cavity for hours and could be detected in some hamsters
future inspection. Cobalt 60 needles (fig. ZC and by counts from cardiac blood during this pe-
D) were also imbedded to provide chronic y riod. An initial leukocytosis following x-irradia-
irradiation. Nonradioactive beads were used as
controls. tion was reported previously for the rabbit and
the chicken by Prosser.8 In the hamster, the
RESULTS latent period was essentially the same (2 to 3
Total body x-irradialion: In hamsters receiv- hours) and the duration similar (12 horn's) for
ing total body x-irradiation with all doses from all doses. This response of the leukocytes was
50 r to 1500 r (100 per cent lethal in nine days), absent in control hamsters observed for com-
the cheek pouch vessels were normal in appear- parable periods of time. Presumably the mobi-
ance, caliber, and contents within the first 2 to lization of circulating leukocytes is a nonspe-
3 hours after irradiation. At that time the leu- cific response to injurious stimulation, in this
kocytes became more numerous, rolling along case x-irradiation, since large numbers are
the endothelium in large numbers (fig. 1C). found in the vessels of the hamster cheek pouch
After 4 to 5 hours the leukocytes were more ad- during infection with Staphylococcits aureus,7 in-
hesive, forming aggregates of 2 to 6 cells and festation with Trichinella,* during malignant
occasionally blocking the lumen of a capillary neoplasia7 and during cortisone administra-
tion.9
* Procured through courtesy of Dr. Walter Kisiel- At 12 to 24 hours, the arterioles were dilated,
eski, Argonne National Laboratory. the circulation in the capillary network was
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relatively active, and the venules distended, to petechial formation was detected on the
producing an erythematous cheek pouch (fig. fourth day after irradiation with 1000 to 1200 r.
ID and E). At this time, the white cells were This altered vascular resistance was correlated
less numerous than in normal non-irradiated with the onset and persistence of vasodilation,
hamsters. Residual leukocytic aggregates were but preceded the erythrocytopenia by 6 to 8
not found. days.
The hematologic values in the total body The blood vessels showed hemorrhagic mani-
x-irradiated hamster have been reported pre- festations within 24 hours after total body
viously.10 The depression in numbers of leuko- x-irradiation, but not immediately. Although
cytes, platelets or erythrocytes as determined maximal at approximately 4 days, the effect
by counts from cardiac blood did not coincide persisted for 21 days. Recovery was usually
in time of appearance with leukocytic adhesive- evident by 30 days. In some hamsters, the
ness to the vascular endothelium, with platelet threshold for petechial susceptibility was as
aggregates immobilized in the peripheral ves- low as 25 r. After lethal x-irradiation, petechial
sels, or with extravasated erythrocytes. Pre- susceptibility as tested by negative pressure de-
sumably the low counts and disappearance of creased strikingly within 1 or 2 days prior to
formed elements in the blood are not due to an death, perhaps because of the hypotension.
accumulation in the peripheral blood vessels or However, a marked increase in petechial sus-
peripheral extravascular tissue spaces in the ceptibility was found in hamsters near ter-
form of petechiae. Injured or worn-out formed minus, by use of the moccasion venom test.
elements are presumably removed by the re- An increase in vasomotor activity was seen
ticulo-endothelial system, especially in the liver occasionally in arterioles at several days after
and spleen. The reduction in numbers is gen- x-irradiation. Vasoconstriction was predomi-
erally conceded to result from destruction of nant near death and related to a concomitant
the hematopoietic precursors," although Furth hyperpotassemia and depressed T-wave of the
and co-workers'2 have presented evidence for a ECG.18
significant loss of erythrocytes by way of the Prior to death of the hamsters (within 24
lymphatic system. In the hamster, a decreased hours) the erythrocytes usually circulated
siLsceptibility to platelet thromboembolism was slowly in aggregates varying from typical rou-
detected by topically-applied thrombin when leaux formation (fig. 2A) to discrete masses or
the platelet count dropped below 100,000 per clusters (fig. 25). In large vessels with more
cu. mm." An extension of the whole blood clot- rapid flow, the erythrocytic aggregates pro-
ting time was found to be correlated with the duced a characteristic granular appearance of
thrombocytopenia and with the disruption of the blood stream. The clusters of red cells re-
perivascular tissue mast cells and decrease in sembled those described by Knisely and co-
tissue histamine content.M No studies were workers17 as "sludged blood" in the mesenteries
made for possible heparin release. of monkeys sick with malaria, except that the
Increased susceptibility to petechial forma- individual elements remained loose in the clus-
tion characterized the first several post-irradia- ter and broke away individually at narrow
tion days, as tested by a negative pressure cup capillary orifices without the formation of oc-
applied to the cheek pouch. Occasionally, a few cluding thrombi. This type of blood flow, char-
spontaneous petechiae were found. In normal acterized as "chunky flow," was typical in
non-irradiated controls, an average of 2 to 4 hamsters near death from heavy total body
petechiae may lie expected at 200 mm Hg nega- irradiation. It may be related to the sedimenta-
tive pressure applied with a standard cup for 1 tion rate, since the occurrence of circulating
minute.16 After irradiation, the number was in- erythrocytic aggregates and increased sedi-
creased to as many as 20 to 30. Intra-pouch in- mentation rates were frequently coincidental in
jections of a standard amount of moccasin the total body x-irradiated hamster.10 It has
venom were also used as a test for hemorrhagic been found in experimental anemia4 and in con-
tendencies. A marked increase in susceptibility ditions characterized by a decrease in circu-
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lating red cell and plasma volume. Erythrocytic definitely significant at all doses by the first
aggregation is a nonspecific phenomenon, since post-irradiation day. The arterioles were wider
it occurs terminally in hamsters after severe by 50 to 100 per cent and the venules were dis-
trauma, urethane poisoning, and overdosage tended. The number of capillaries with an ac-
with steroids.9 Although it may be similar to tive circulation were increased. The vasodila-
the condition called "sludged blood," we have tion persisted for at least 8 days in the pouches
not used this term because of the prognostic irradiated with 25 to 1000 r and until necrosis
implications of the phrase. We think that the and healing by 30 days in the higher dose range.
concept of "sludged blood" should be modified Both arterioles and venules became varicose
with less emphasis on causative attributes and or tortuous in appearance at 24 hours after ir-
that the term, should be redefined. radiation of the pouch with 15,000 r and above
After total body x-irradiation, hamsters were (7 of 8 hamsters) and at 48 hours with doses
more susceptible to infection as shown by local less than 15,000 r (fig. 2E). Occasional arte-
invasions at previous pin holes in the cheek rioles were constricted locally, producing a
pouch incidental to eversion and spreading for sausage-shaped segment (fig. 2F). A few pete-
transillumination. Normally the hamster is chiae were found spontaneously after irradia-
highly resistant to infection and the same cheek tion in most of the hamsters at all periods of
pouch may be pinned out and examined many examination. An increase occurred in suscepti-
times without infection. When infection was bility to petechial formation resulting from
present in the cheek pouch, white cells were negative pressure applied to the x-irradiated
numerous, adhering to the endothelium. En- pouch and in some instances the non-irradiated
teric microorganisms (e.g., E. coli) were found contralateral cheek pouch.
in the blood at 7 days after 1200 r. Presumably In pouches exposed to 10,000 r and more, the
the leukopenia and injury to bone marrow and irradiated area became an erythematous spot
spleen result in a lowered resistance, and per- detectable by gross inspection on the 7th to
haps the mucosa and its vessels are rendered 11th days. At this time, the vascularity was
susceptible. Enteric infection may be a con- greatly increased especially at the rim of the
tributory cause of death in heavily irradiated exposed tissue. Vast numbers of minute, tor-
hamsters. tuous and looped capillaries formed extensive
Cheek pouch x-irradiation: Using 54 ham- networks at the margin of the irradiated area.
sters, a small portion of the cheek pouch (10.0 Minute spontaneous petechiae (fig. 2E) were
mm. diameter) was irradiated with 25 r to relatively numerous at the venous junctions.
30,000 r, while the rest of the pouch and animal The abundance, distribution, and character-
were shielded (fig. IB). The irradiated spot was istic morphology of the tiny capillaries appear-
marked for daily study by a non-irritating ing at this time can be explained adequately
glass bead imbedded in the pouch seven days only as the result of growth of new blood ves-
previously (fig. 2C and D). sels. The number of pre-existing non-circulat-
Vasodilation was produced by all doses (25 r ing capillaries which become active as a result
to 25,000 r). The entire pouch was involved of arteriolar vasodilation cannot account for
without limitation of the response to the spot the vast increase in vascularity. Presumably
irradiated (fig. 2 C and D). The vasodilation the endothelium responds to x-ray injury by
was detected by inspection and measurement of reparative growth in a non-specific way com-
vessel diameters in successive photographs of parable to that following thermal burns, physi-
the same cheek pouch taken before irradiation, cal trauma, and infection.19
within 5 minutes of irradiation, and at 2, 3, 7, Considerable edema occurred in the entire
14 and 30 day intervals. No vasodilation was pouch from the 2nd to the 11th day after doses
found in control hamsters which were handled of 5,000 to 25,000 r. The epithelial tissue
and photographed in the same way. The thresh- sloughed off and necrosis developed with 10,000
old dose for the response was 20 r. The response r and more. Necrosis occurred at 5,000 r when
was slight immediately after irradiation but the entire cheek pouch was irradiated. The
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necrotic response was confined to the irradiated the cheek exterior to the normal location of the
spot and became evident at 7 to 15 days, fol- cheek pouch within the buccal cavity, and the
lowing increased vascularity and petechial for- skin at this point foi med a black scab-like crust.
mation. Within 30 days, all spots were healed, In a few hamsters, the bead was left in the
with marked puckering of the area. Xecrotic pouch and remained in position for periods as
tissue was shed and the area was free of infec- long as 14 days. Marked necrosis in the pouch,
tions. considerable pus formation, and adhesions were
Imbedded radioactive fragments: Sr90 beads characteristic in such experiments. The ham-
(approximately 1 mm. in diameter) and Y91 sters lost weight, became scrawny and exhibited
beads with activities of 140, 76, 50, 25, 10 and chronic diarrhea. Most of the hamsters sur-
1 /uc were inserted into the connective tissue of vived, although a few did not. At 35 days the
the cheek pouch and studied at intervals of 24 lesion in the cheek pouch was partly healed.
hours, 48 hours, 72 hours, 6 days, 15 days and The blood vessels and the circulation were
30 days after implantation. A total of 70 ham- studied at magnifications of 200 to 500 X in the
sters were used. The cheek pouch containing area exposed to Sr9u glass beads. With high ac-
an imbedded bead was reinserted in the buccal tivity beads, platelet thromboembolism was
cavity. Nonradioactive beads were imbedded prevalent in the venules. Leukocytic aggregates
similarly for use as controls. Representative were numerous on the endothelial lining of
hamsters were injected with intravascularly venules. The blood vessels were fragile as .«hown
precipitated lead chromate, and the cheek by numerous petechial hemorrhages which
pouches were excised and prepared as whole formed at branching points on the venous side
mounts for study of the vascularization. of the capillary network. Petechiae were not
At 24 hours, the high activity beads (140 MC0 found on the capillaries per se. The arterioles
were encircled by a ring of dense minute capil- were frequently narrow, especially for short
lary networks indicative of endothelial prolif- portions, producing "sausage-shaped" seg-
eration (fig. 35). The newly-formed capillaries ments. Asynchronous fibrillation of the indi-
were short and characterized by tiny loops. vidual skeletal muscle fibers of the pouch was
Pre-existing arterioles and venules developed observed and recorded on motion picture film.
a marked tortuosity. The entire cheek pouch Low activity beads (1 ^c.) produced a slight
was erythematous as a result of vasodilation increase in vascularity by 30 days, but severe
and edema. No such changes in vascularity effects were not observed even with the bead
were seen in the vicinity of non-radioactive implanted for more than 3 months. Activities
glass beads. Some petechial hemorrhages were of 25 or 10 ^c. produced considerable vascu-
observed at venous junctions in the vicinity of lar effects and the typical twitching phenome-
the radioactive bead. At 48 hours, the epi- non characteristic of damaged skeletal muscle
thelium overlying the bead was opaque, white fibers. The severity of the effects was related
and blister-like, flaking off in scales when to the amount of radioactivity. Furthermore
touched with forceps. The diameter of the blis- the time of onset of the vascular effects was
ter was approximately double that of the bead. much earlier for high activities.
An extremely vascular rim composed of very The qualitative effects of Co80 needles (fig.
fine capillary networks bordered the periphery dC and D) were similar to those produced by
of the blister. The beads were usually removed Sr90 glass beads. However, the vascular changes
from the cheek pouch at 3 or 4 days. If not developed more slowly and tissue necrosis Avas
removed at this time, the beads frequently much less extensive with comparable doses of
eroded through the epithelium and were usu- Co60.
ally recovered within a few days impacted in a A marked increase in susceptibility to pete-
bit of feces in the bottom of the cage. Subse- chial formation was detected by the application
quent observations were made and infections of negative pressure to the cheek pouch con-
were frequent in the necrotic area. At 2 to 3 taining radioactive fragments. In some in-
weeks, the hair was often lost from a patch on stances involving high activity Sr90 beads in the
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cheek pouch or back, the blood vessels of the for increased susceptibility to petechial forma-
non-irradiated contralateral pouch developed tion does not seem to be thrombocytopenia.
increased numbers of petechiae in response to Furthermore, x-irradiation of either the entire
the negative pressure test. The bleeding time cheek pouch or portions, with the rest of the
and resistance to local infection appeared to be hamster shielded, produced petechiae in the
altered in the contralateral cheek pouch as well area irradiated, but without significant changes
as the one irradiated. Although these observa- in the blood count. On the other hand, both
tions suggest the possibility of a circulating hu- spontaneous petechial formation and increased
moral agent resulting from radiation injury, susceptibility have been obtained in hamsters
the results of further experiments in progress with thrombocytopenia induced by immuno-
are necessary before coming to a definite con- logical procedures. The increased petechial
clusion. susceptibility may be related to the disruption
of mast cells, demonstrated in whole mounts
DISCUSSION stained metachromatically with toluidine blue
In general the vascular effects of x-irradia- or methylene blue, resulting in probable re-
tion in the hamster are in agreement qualita- lease of histamine as shown by a decrease after
tively with the post-irradiation findings re- irradiation in histamine content of cheek
ported by Smith, Svihla and Patt2 in the wing pouches and mesenteries.M Release of heparin
of the bat. A major difference is the lack of from disrupted mast cells is another possible
hcmorrhagic effect in the bat, even with the factor, although the increase in petechial sus-
highest doses used. In fact, the bat was found to ceptibility precedes the onset of significant pro-
be highly resistant to x-irradiation, requiring longation of the blood clotting time. Irradia-
15,000 to 60,000 r delivered to the entire body tion seems to alter some component of the blood
to shorten the life span and 10,000 to 60,000 r vessel wall or perivascular sheath, either by
to produce vascular changes. Furthermore, direct action or indirectly by production of a
doses in excess of 50,000 r delivered to portions humoral agent. Adrenalectomy prior to total
of the wing membrane were required for circu- body x-irradiation does not prevent post-irra-
latory effects. In the hamster, the LDW/M was diation petechial formation.
determined as 880 ± 12 r,10 and doses of 1500 r The precise mechanism involved in petechial
were found to be 100 per cent lethal in 9 days. formation resulting from x-irradiation or other
The circulatory changes in the hamster oc- types of trauma is still not known. Possible fac-
curred at much lower doses than those reported tors are an alteration in the integrity of the
in the bat. Consequently, the vascular changes perivascular sheath, retraction or realignment
in the hamster are probably more nearly typical of the interlocking processes of the vascular
of effects in man and other commonly-studied endothelium, and erosion of the interendothe-
mammals. Species differences in hemorrhagic lial cement substance. We have studied the
effects of x-ray exposure have been reported by actual formation of petechiae in the hamster
Rosenthal and Benedek, with greater re- cheek pouch, using moccasin venom as the
sponses in man, dog and guinea pig as com- traumatic agent for consistency of widespread
pared with rat, mouse and particularly rabbit. petechiation and ease of continuous microscopic
Although an evaluation of the factors which observation and motion picture photography
cause bleeding after x-irradiation is still diffi- in limited fields at high magnification. The
cult, certain possibilities are more probable erythrocytes spurted "one by one" through the
than others. The hemorrhagic tendency de- vessel wall, primarily at venous junctions, but
velops within the first several post-irradiation without detectable rupture of the wall. Further-
days during the period of decreased total white more, cessation of petechiation was not accom-
cell count and reversal in the polymorpho- panied by either fibrin clot formation or devel-
nuclear leukocyte to lymphocyte ratio, but opment of a platelet plug.
prior to the period of decreased circulating A number of substances have been investi-
blood platelets. Consequently, the explanation gated for possible value in the prevention of
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post-irradiation petechiae or susceptibility to doses, erythrocytes usually circulated in aggre-
petechial formation as tested in the hamster gates varying from typical rouleau formation
cheek pouch by negative pressure or moccasin to discrete masses, resembling "sludged blood"
venom. Pre-irradiation treatment with Adreno- but without formation of occluding thrombi.
sem reduced the hemorrhagic effects of total Partial cheek pouch x-irradiation with 25 r to
body x-irradiation.11 Rutin, administered prior 25,000 r produced vasodilation of the entire
to irradiation, was somewhat effective as shown pouch at 24 hours, persisting for at least eight
by the moccasin venom test. days at lower doses and until necrosis and heal-
The increase in bleeding tendency per se does ing by 30 days with higher doses. Tortuosities
not appear to be a major factor contributing to appeared in arterioles, venules and capillaries,
death in heavily x-irradiated hamsters. The particularly with 10,000 r or more. Occasional
erythrocytic aggregates are probably more arterioles were constricted locally, producing
symptomatic than causative. Changes in the "sausage-shaped" segments. Vast numbers of
ECG, blood pressure and electrolyte imbalance minute, looped and newly-formed capillaries
are probably secondary. Although the cause of developed at the margins of the irradiated
death is not known, predilection to enteric in- spots. Tiny petechiae were relatively numerous,
fection as a result of lowered resistance from predominantly at the comparatively vulnerable
irreversible injury of bone marrow and spleen venous junctions. Within 30 days, all irradiated
may well be an important factor.21 As suggested spots were healed, with residual puckering of
by Cronkite and co-workers,1 ulcero-necrotic the area.
lesions and infections initiate hemorrhage Chronic vascular effects of irradiation were
which may become extensive because of the studied at periodic intervals for 30 days in
thrombocytopenia. Consequently, hemorrhage cheek pouches containing Sr90 glass beads and
may still be an important factor operating in- Co60 needles, imbedded in the subepithelial con-
directly. nective tissue of the pouch. A rim of dense
minute capillary networks developed around
SUMMARY the radioactive fragments. Pre-existing arte-
The changes in blood circulation in the ham- rioles and venules acquired extensive tortu-
ster cheek pouch were studied by direct obser- osity. Vasodilation was prevalent. Platelet
vation and cinephotomicrographic technics thromboembolism, "sausage-shaped" arteriolar
after single doses of total body x-irradia- segments, petechial hemorrhages at venous
tion, whole or partial cheek pouch x-irradiation junctions, edema, and marked tissue necrosis
and chronic exposure of the cheek pouch to im- occurred in varying degree depending upon the
bedded radioactive fragments (/3-emitting dosage. Asynchronous fibrillation occurred in
strontium glass beads and 7-ray cobalt needles). individual skeletal muscle fibers of the pouch.
Transient leukocytosis and leukocytic ad- High activity beads (140 NC.) produced severe
hesiveness to venular endothelium occurred at vascular and tissue damage, frequently eroding
4 to 12 hours after x-irradiation of the entire through the pouch epithelium in 3 to 7 days
body with doses from 50 r to 1500 r (100 per and predisposing to local cheek pouch infec-
cent lethal in nine days). Subsequently, vaso- tions. The vascular changes with Co60 needles
dilation occurred in arterioles and venules, ac- developed more slowly and tissue necrosis was
companied by increased blood flow in the capil- much less extensive than with comparable doses
laries. Circulating leukocytes were decreased in delivered by Sr90 glass beads.
numbers. Susceptibility to petechial formation The causative factors and mechanisms in-
was increased as tested by negative pressure volved in the hemorrhagic effects of ionizing
and intrapouch injections of moccasin venom. radiation are discussed, and an attempt is made
Near death, pancytopenia and vasoconstriction to evaluate the relative importance of thrombo-
were prevalent and associated with concomi- cytopenia, prolonged whole blood clotting time,
tant hyperpotassemia and depressed T-com- decrease in numbers and increase in disruption
ponent of the ECG. Prior to death from lethal of perivascular tissue mast cells and direct
Downloaded from http://circres.ahajournals.org/ by guest on September 23, 2015142 EFFECTS OF IRRADIATIONS ON CIRCULATION
effect on the blood vessels. A component of the Roentgeno-irradiation partial del sacculo con
blood vessel wall or perivascular sheath ap- 25 a 25.000 r produceva vasodilatation del
peal's to be altered by x-irradiation acting either integre sacculo post 24 horas. Con le plus basse
directly or indirectly by means of a humoral doses intra le limites mentionate le vasodilata-
\igent produced by injury. tion persisteva dura n te al minus octo dies;
con le plus alte doses illo persisteva usque al
ADDENDUAT adventimento de necrosis e curation intra 30
Thefindingspresented in this paper are illustrated dies. Tortuositates appareva in arteriolas,
in a Kodaehrome motion picture film entitled venulas, e capillares, specialmente con irra-
"Effects of Beta and X-irradiation on the Circula- diationes de 10.000 r o plus. Occurreva in-
tion in the Hamster Cheek Pouch" (16 mm, 400 feet, frequente constrictiones local del arteriolas,
15 minutes, silent with titles). The photographs resultante in segmentos a "forma de salsicia."
were taken by Frederick W. Maynard.
Al margine del areas irradiate, grande numeros
ACKNOWLEDGMENT
de minuscule capillares ansiforme esseva
novemente disveloppate. Parvissime petechias
The authors acknowledge with thanks the assist-
ance of David L. Joftes, Ph.D. in the initial phase esseva relativemente numerose. Illos pre-
of the work. dominava al comparativemente vulnerabilc
junctiones venose. Intra 30 dies omne le areas
SUMMARIO IN INTERLINGUA irradiate esseva curate. Illos remaneva marcate
Le alterationes del circulation sanguinee in le per rugas residue.
sacculo buccal del hamster esseva studiate per Chronic effectos vascular de irradiation
observation directe e per technicas cine- esseva studiate a intervallos periodic, durante
photomicrographic post (1) roentgeno-irradia- un periodo de 30 dies, in le sacculos continente
tion del corpore total, (2) roentgeno-irradiation perlas de vitro a Sr90 e agulias de Co80, in-
partial o integre del sacculo, e (3) exposition fundate in le subepithelial histos conjunctive.
chronic del sacculo a infundate fragmentos Un margine de minute reticulos capillar sc
radioactive (perlas de vitro strontial a emana- disveloppava circa le fragmentos radioactive.
tion /3 e agulias cobaltic a radios 7). Pre-existente arteriolas e venulas disveloppava
Transients leucocytosis e adhesivitate leuco- extense tortuositates. Vasodilatation esseva
cytic al endothelio venular se manifestava 4 a prevalente. Thromboembolisino a plachettas,
12 horas post roentgeno-irradiation del corpore segmentos arteriolar a "forma de salsicia,"
integre con doses de 50 a 1500 r (effectuante hemorrhagias petechial a junctiones venose,
un mortalitate de 100 pro cento post 9 dies). edema, e marcate histonecrosis occurreva a
Subsequentemente il occurreva vasodilatation varie grados secundo le dosage administrate.
del arteriolas e venulas, accompaniate per un Fibrillation asynchrone occurreva in individual
augmento del fluxo sanguinee in le capillares. fibras skeleto-muscular del sacculo. Perlas a
Le conti'o del leucocytos circulante esseva alte activitate (140 MC) produceva sever damnos
reducitc. Le susceptibilitate al formation de vascular e histolesiones, frequentemente con
petechias se monstrava augmentate in tests a erosion a transverso le epithelio saccular intra 3
pression negative e in injectiones intrasaccular a 7 dies e con consequente predisposition a
de veneno de serpentes agkistrodontal. Mori- infectiones local del sacculo. In le caso dc
bunditate, pancytopenia, e vasoconstriction agulias a Co60 le alterationes vascular se dis-
esseva prevalente e associate con simultanee veloppava plus lentemente, e le histonecrosis
hj'perkalemia e deprimite componentes T del esseva multo minus extense que in le caso de
electrocardiogrammas. Ante morte ab doses comparabile doses emanate per perlas de vitro
letal, erythrocytos circulava usualmente in a Sr90.
aggregatos variante in forma ab typic rouleaus Le factores causal e le mechanismos in-
usque a massas discrete resimilante "sanguine volvite in le effectos de radiation ionisante es
fangose" sed sin formation de thrombos oc- discutite. Es facite le tentativa de evalutar le
cludente. importantia relative de thrombocytopenia,
Downloaded from http://circres.ahajournals.org/ by guest on September 23, 2015FULTON, LUTZ AND KAGAN 143
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Downloaded from http://circres.ahajournals.org/ by guest on September 23, 2015Effect of X-irradiation and Beta Emanation on Circulation in the Hamster Cheek Pouch
GEORGE P. FULTON, BRENTON R. LUTZ and ROMA KAGAN
Circ Res. 1956;4:133-143
doi: 10.1161/01.RES.4.2.133
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