Experimental Friction Blisters in Macaque Monkeys*

Vol. 53, No. 2 PriNted in U.S.A.

THE JOURNAL OW INVESTIGATIVE DERMATOLOGY

Copyright

1969 by The Williams & Wilkins Co.

EXPERIMENTAL FRICTION BLISTERS IN MACAQUE MONKEYS' THOMAS A. CORTESE, JR$ MD., PrnD., TOMMY B. GRIFFIN, MAJ. MC, LAURENCE L. LAYTON, PHD.t AND THOMAS C. HUTSELL, PHD. ABSTRACT 1. Friction blisters were successfully produced experimentally on the palms and soles of two Macaca ins and four Macaca speciosa monkeys. The lesions corresponded to those seen clinically and experimentally in man.

2. Histologically, monkey friction blisters represented intraepidermal vesicles and were identical to human friction blisters. 3. Monkey friction blister fluid qualitatively reflected the composition of serum. 4. The production of experimental friction blisters in laboratory animals and the close resemblance of these lesions to those of man will allow investigations of frictional injury which would be difficult or impossible to perform in man.

The pathogenesis and me contents of friction

blisters in man have recently been studied (1—4). To our knowledge there have been no investigations or reports of friction blisters or

METHODS AND MATERIALS

Experimental animal.s and their handling. Two species of Old World monkeys, the Macaca irus (Cynomolgus, closely related to the rhesus) and

their experimental production on the skin of the Macaca speciosus (Stumptail) were selected for

these studies. A total of 6 monkeys, 2 Macaca ins

non-human species.

Although the diseases of the human integument nre best studied in man himself, the production of the same lesions in the laboratory animal affords an opportunity to study a vari-

ety of diseases which would be difficult or hazardous to produce and investigate in the human volunteer. This study reports the successful accomplishment of the experimental production of friction blisters on the skin of monkeys.

and 4 Macaca speciosa, were used.

The animals were tranquilized with injectable Acepromazine Maleate (Ayerst Laboratories, Inc.,

New York, N. Y.), 0.5 mg per pound of body weight. With this amount of drug the animals remained tranquil from 1 to 3 hours. They were then

fastened to an animal board with extremeties outstretched and held in place for the blistering procedure.

Friction blisters. Experimental friction blisters ware produced on the palms and soles of each monkey by the pencil eraser, twist rubbing technique (2).

Histology. Using 1% lidocaine hydrochloride for

The authors wish to acknowledge Mr. Paul S. local anesthesia, 4 millimeter punch biopsies were Peyakovich for his able assistance with the blister- taken from 10 representative blisters immediately ing procedures on the monkeys and Mr. Harry and 48 hours after blistering. Each specimen was Roberts of the Letterman General Hospital Photo prepared by routine methods and stained with Laboratory for his very fine photomicrographs. Reprint requests to Dermatology Research Di- haematoxylin and eosin and with periodic acidvision, Letterman Army Institute of Research, San Schiff stains for light microscopy examination. Francisco, Califomia 94129 (Maj. Griffin). Blister fluid and corresponding serum. Blister In conducting the research described in this re- fluid and corresponding serum were obtained from port, the investigators adhered to the "Guide for 2 Macaca speciosa monkeys 4 hours after blistering. Laboratory Animal Facilities and Care," as pro- Monkey serum was obtained by vena puncture of mulgated by the Committee on the Guide for the saphenous vein. Blister fluid was aspirated from Laboratory Animal Resources, National Academy of Sciences—National Research Council.

Received March 10, 1969; accepted for publica-

tion March 31, 1969.

* From the Dermatology Research Division, Letterman Army Institute of Research, San Francisco, California 94129.

each blister using a 26 gauge needle and a tuberculin syringe. The needle was inserted into the blister

cavity approximately halfway between its center and its edge. The skin overlying each blister was cleansed with 70% isopropyl alcohol and allowed to

t Present address: Department of Dermatology, air dry just prior to aspiration. The collected blister Indiana University Medical Center, 1100 West fluid was pooled for each monkey and submitted along with the corresponding serum specimens for Michigan Street, Indianapolis, Indiana 46207. t Western Regional Research Laboratory, Al- biochemical analysis. Biochemical analyses. Sodium and potassium bany, California 94710. § Present address: Department of Biochemistry, concentrations were analyzed by flame photometry. G. D. Searle Laboratory, Chicago, Illinois. Calcium values were determined using a Turner 172

FRICTION BLISTERS IN MACAQUE MONKEYS

Fluorometer according to the method published in the G. K. Turner Fluorometer manual. Total protein was determined by the method of Lowry, et at. (5). Zone electrophoresis was carried out on a Beckman Microzone Electrophoresis Apparatus, Model R-lO1, using cellulose acetate as the supporting medium according to the method of Grunbaum, et at. (6). All analyses were done in duplicate and verified against appropriate controls. RESULTS

173

obtained from each blister, which necessitated the pooling of specimens for biochemical analysis.

Except for the slower blister filling time of 2 to 4 hours (in man blister filling requires only 1

to 2 hours (2)), all of these events in the formation of a clinical friction blister duplicated our observations in human volunteers. It is possible that the slower blister filling time in mon-

keys resulted from their supine positions and Blister formation,. A total of 96 friction blisrestraints, as 3 to 4 hours are required for ters were made on the palms and soles of the friction blisters to fill in man when they are 6 monkeys (Fig. 1). As many as 12 friction blis-

ters could be produced on one palm or sole of the Macaca speciosa monkeys. The animals gave no signs that they experienced any substantial degree of pain or discomfort from the blistering procedure. The time required to produce the blisters varied from 45 seconds

to 5 minutes of twist rubbing. During twist rubbing, minimal erythema of the rubbed site could be seen. At the moment of blistering, a narrow zone of loosened skin appeared at the periphery and gradually increased toward the center of the rubbed site until the blister top was formed and elevated over the underlying skin. Within 2 to 4 hours after blistering, the blister was filled with a clear, colorless to pale yellow fluid. Only about 0.05 ml of fluid was

made with the volunteer's hand held straight above his head and held there for 1 hour (2). Histology. Immediately after blistering

trauma, a cleft was observed within the malphigbian layer, as shown in Figures 2 and 3. The blister roof was composed of the stratum corneum, stratum granulosum and both normal and damaged prickle cells. The blister base consisted variably of normal appearing basal and prickle cells with superimposed distorted or destroyed prickle cells. The cellular

material lining the blister cavity was often amorphous and devoid of nuclei. There was usually cell preservation in the rete ridge portions with marked reduction or total absence of cells and only cellular debris over the dermal

Fio. 1. Sole of a Macaca speciosus monkey showing ten friction blisters two hours after blistering trauma. Darkened areas within the blister outlines are a result of blister fluid accumulation. Two healed, but still scaling, two week old friction blisters, can be seen in the center.

174

THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

Pie. 2. Photomicrograph of a frictien blister from the sole of a Macam speeiosus monkey

immediately after blistering trauma. Note the mid-malphigbian location of the blister cleft. Compare with the human friction blister, Fig 4. II & E stain. X 100

-1

•

• ..

'a' " S, flr", 4 b ——

&

4.

1_• —T

—

S S

.9 ,

a

FIG. 3. Higher magnification of Fig 2 showing the edge of the blister cavity. Note partIcularly the elongation, distortion and destruction of the spinous cells. Compare with the human friction blister, Fig 5. H & E stain. )< 250

175

FIG.

1

I

V..—

4.

A. -

•-e

•

4.

t.'.. 'I..

t

—

FRICTION BLISTERS IN MACAQUE MONKEYS

4. Photornicrograph of a friction blister from the palm of a male volunteer imme-

diately after blistering trauma. Compare with Fig 2.11 & E stain. X 100

papillae. Only rarely was disruption of the blister fluid qualitatively reflects the composibasement membrane noted. The dermis lacked tion of serum (4).

an inflammatory infiltrate and the dermal vessels and appendages remained intact and

DISCUSSION

To our knowledge the questions whether Thus, not only clinically but histologically friction blisters are clinically experienced by the experimental friction blister produced on species other than man and whether such blisnon-human primate skin appeared to be the ters can be experimentally produced had not unaltered.

same as that produced on human skin. The loca- been answered. This report represents the first tion of the blister cleft and the cellular changes investigative attempt to show that vesicles of

in both blister roof and blister base were prac- the friction type can be experimentally protically identical to those in man (2, 3) (Figures duced on the palms or soles of at least two dif4 and 5). The healing pattern observed in the ferent species of Macaca monkeys. 48 hour biopsies was also similar.

Blister fluid. The data in Table I present the total protein, albumin, sodium, potassium and calcium concentrations in blister fluid and corresponding serum from two Macaca speciosa

The Macaca speciosa monkeys proved to be the better species on which to produce friction blisters. Their palmar and plantar skin more closely resembled the skin of the same sites in man—in texture, consistency, and

monkeys. The values obtained qualitatively color—and also possessed all of the necessary indicate that the blister fluid constituents ana- prerequisites for blistering (2). As in man, the lyzed corresponded to those present in serum. blistering end point was readily perceptible. Because the analyses were performed on speci- The palmar and plantar skin of the Macaca mens from only two monkeys, it is not possible

irus monkeys possessed a thinner, more delicate

to ascribe any statistical significance to these stratum corneum and was more pliable and values. The determinations, however, are in ac- less rigidly held to the underlying structures, cord with the human studies that friction thus greater care was required to achieve blis-

THE JOIJENAL OF INVESTIGATIVE DERMATOLOGY

176

—

;

a

FIG. 5. Higher magnification of Fig 4 showing the edge of the blister cavity. Compare

with Fig 3. H & E stain. X 250

TABLE I

tering so as not to produce abrasions. The

Blister fluid and corresponding total serum protein, albumin and electrolyte concentrations from two Macaca speciosa monkeys

blistering end point was also more difficult to

Monkey 51 Serum

Sodium (mEq/L)

Potassium (mEq/L) Calcium (mg/

152

(gin/100 ml)

Albumin (gm/ 100 ml)

157

Serum

149

mister 164

5.2

4.1

5.2

10.4

9.1

10.6

9.0

8.1

6.7

8.4

6.5

4.1

2.6

(50%)" (38%)

tion. Gross and microscopic appearances confirmed

that friction blisters on monkey skin correspond

to those seen clinically and experimentally in man. The blister contents analyzed were likewise

4.1

100 ml)

Total Protein

blister

Monkey 52

discern because of their skin's greater pigmenta-

similar to those of man and qualitatively reflected the composition of serum.

Monkeys can, therefore, serve as suitable laboratory animals for the experimental production and study of friction blisters. As with

other laboratory models, certain studies of frictional injury can now be undertaken which would be difficult to perform in man.

4.3 (51%)

2.3 (36%)

a The values represent duplicate analyses on

pooled friction blister fluid obtained 4 hours after blistering trauma from 24 friction blisters on the soles of each monkey. b Percentage values were obtained by densitom-

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FRICTION BLISTERS IN MACAQTJE MONKEYS

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