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Responses of the Human Eccrine Sweat Duct to Controlled Injury
RESPONSES OF THE HUMAN ECCRINE SWEAT DUCT TO CONTROLLED INJURY GROWTH CENTER OF THE "EPIDERMAL SWEAT DUCT UNIT*
WALTER C. LOBITZ, JR., M.D., JOHN B. HOLYOKE, M.D. AND WILLIAM MONTAGNA, Pn.D.
The human ecerine sweat duct may be divided into two parts. [1] The "dermal
sweat duct" (that portion that lies within the dermis) and [2] the "epidermal sweat duct unit" (that portion that lies within the epidermis). The "dermal duct" is composed of two layers of cells. [1] The basal cells which rest on the connective tissue sheath are usually euboidal and contain occasional mitotic figures. [2] The "lumenal cells" which line the lumen of the sweat duet are euboidal or columnar, have a hyalinized lumenal border that varies in size, and are never seen to contain mitotie figures. The "Epidermal Sweat Duct Unit" (1) is composed of three major parts. [1] The intraepidermal portion of the sweat duet which is made up of the single layer of cells lining the lumen and is a continuation of the lumenal cells of the dermal duet. These lumenal or lining cells are euboidal or rectangularly flat when arranged about the lumen. They do not have priekies between themselves, they have a clear cytoplasm in the lo\ver two-thirds of the epidermis, but accumulate "Keratohyalin" granules in the outer one-third of the epidermis. [2] The periduetal sheath of two or more epidermal (prickle) cells that are concentrically arranged around the lumenal cells. These sheath cells form a mass of cells within the epidermis. About the apex of this cell mass are pigment-free basal cells that are continuous with the basal cells of the dermal duet on the one side, and with the basal cells of the adjacent epidermis on the other. The flat end of the cell mass, in the center of which is the sweat pore, is on the surface of the skin. [3] The sweat pore (the ecerine sweat duct unit opening on the surface of the skin)
is encircled by a ring of keratin fibers (O'Brien's Keratin Ring (2)) that are, adjacent to the keratin fibers of the stratum corneum. Within the keratin ring and lining the lumen of the pore, is a ring of Sehiff-positive material (not glyeogen) that contains less disulfide concentrations than the keratin ring and is continuous with a similar Sehiff -positive material that covers the free surface of all of the lumenal cells of the entire dermal and epidermal eeerine sweat duet. The histoehemieal contents of these various cells in the dermal and epidermal
portions of the human eeerine sweat duet have been described and add more * From the Department of Dermatology (W.C.L., Jr.) and the Department of Pathology (J.B.H.) Hitchcock Clinic and Dartmouth Medical School, Hanover, New Hampshire;
and the Arnold Biological Laboratory (W.M.) Brown University, Providence, Rhode Island.
This study was supported by the Quartermaster Research and Development Command (Contract ,DA44-1O9 q.m. 1424) and by the Hitchcock Foundation.
Presented at the fifteenth Annual Meeting of The Society for Investigative Dermatology, Inc., San Francisco, California, June 19, 1954. 329
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evidence to the similarities, differences, and "biological structure" of their component parts (1, 3—7).
Hoepke (8) was among the first to point out that the sweat duct in the epi-
dermis was a closed tube of lining cells and not merely a canal-like space bet\veen
prickle cells. However, "he insists that the wall is but a functional entity and that its building stones are the prickle cells of the epidermis." (9) Pinkus (9) reviewed this and added evidence to a concept that the epidermal portion of the sweat duct behaved independently and differently from the rest of the epidermis. Among other things he pointed out that in the epidermis: [1] The duct was un-
affected by the surrounding edema and vesiculation in experimental allergic dermatitis. [2] The duct retained its normal structure even \vhen surrounded by the dyskeratotic cells of senile keratoses and the anaplastic cells of Bowen's disease. [3] There are no melanin granules in the duct cells of normal Negro or hyperpigmented white skin, nor in the stratum corneum overlying the sweat duct. (We have never seen pigment in the basal cells at the apex of the cone of the "epidermal sweat duct unit" even though the adjacent basal cells are heavily laden with such pigment.) [4] The epidermal sweat duct spirals to accommodate its length to the adjacent epidermis; it is straight in the acanthotic epidermis of psoriasis and tightly coiled when the epidermis is narrowed with atrophy. Pinkus then raised the question as to where the "matrix for the replacement of these cells could be". He postulated, on the evidence then at hand, that the entire length of the sweat duct might serve as the matrix by mitotic cell division. He states that histopathologically "adenomatous proliferation of the syringeal
epithelium is not uncommonly found near lupus vulgaris, after roentgen-ray treatment of carcinomas and in other chronic processes which destroy the upper ends of the ducts"; and "it is a long recognized fact that remnants of the appendages help materially in the epithelialization of granulating wounds". Such types of questions are always pertinent to those interested in normal and abnormal growth patterns in the human. Questions concerning the ecerine sweat duct are of special interest to those focusing their attention on the structure and the function of the human eccrine sweat gland and on the sweat retention
group of diseases in particular. For such reasons, histologic studies (10) were made and confirmed some of Pinkus' observations. Recent histochemieal studies added more evidence that the "Epidermal Sweat Duct Unit" (1) is a biologic as well as a morphologic entity that behaves differently from the epidermis and as such must be further investigated. The following observations have resulted from some of our attempts to better understand this ecerine sweat duct in the human. PROCEDURE
This report will describe the effects of t\vo different types of injury to the epidermis and to the ecerine sweat duet on the growth responses of the sweat duet. The effect of such injury to the growth of the epidermis and to the pilosebaceous units will be described at a later date. The injuries to be reported in this paper are: [1] "The Strip Experiment":
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Removal
of the stratum corneum of the epidermis by the Strip Method of
Pinkus (11) [2] "The Burr Experiment": The removal of the entire epidermis including the "Epidermal Sweat Duct Unit" with a high-speed rotary driven abrasive. Young adults, human males, were utilized in these studies. The areas injured were on the skin of the back in the sites of predilection for the sweat retention group of diseases and where the sweat gland population count was known to be high.
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All biopsies were taken without local anesthesia using the highspeed motor driven rotary 5 mm. punch (12). For this report the specimens were fixed immediately in chilled ilelley's solution. The tissue sections were stained with [1] hematoxylin and phloxine [2] Schiff reagent (13) with and without hematoxylin as a eounterstain and controlled with saliva for the removal of glycogen [3] Toluidine Blue, buffered at pH 5, and controlled with ribonuelease.
FIG. 1. The "Epidermal Sweat Duct Unit" immediately after stripping. The swollen basal cells of the adjacent epidermis sharply demarcate the unaltered "unit" (McManus procedure counterstained with hematoxylin) Mag. X 410.
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The Strip Experiment Seven young adult males were utilized. The area for study selected was the skin of the back between the lateral markings of the scapuli and posterior axillary
fold. A linear area of stratum corneum was stripped off with scotch tape. In order to get consistent stripping and response from subject to subject and during
repeated experiments in the same subject, the epidermis was stripped until moisture appeared on the skin. To accomplish this, for example, from 16—24 strippings were necessary on comparable areas of skin even in the same individual. Biopsies taken after stripping were as follows: [1] one group of men 16, 24, 48,
and 72 hours after stripping; [2] one group of three men 3, 5, 10, and 15 days after stripping; [3] one man immediately after stripping (approximately 5 minutes from the onset of injury), 4, 8, 16, 24, 48, and 72 hours and 5, 10, and 15 days after stripping.
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A most striking change occurred in the biopsy taken immediately after the injury. In the general epidermis the basal cells became swollen to almost t\vice the original diameter; the cytoplasm was clear and completely devoid of any stain or counterstain (14). The basal cells of the "Epidermal Sweat Duct Unit," on the other hand, did not undergo any such swelling (Fig. 1). Such swollen adjacent basal cells of the epidermis demarcate the "Epidermal Sweat Duct Unit" in even greater contrast. In addition the glycogen accumulation and utilization that occurred over the next 5 days after injury, did not take place in the respective
FIG. 2. The conical shaped fine abrasive used to burr off the epidermis.
RESPONSES OF HUMAN SWEAT DUCT
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cells of the "Epidermal Sweat Duet Unit" at any time, nor was there any burst of mitotie activity in these cells. The Burr Experiment
Three adult males were used. The sweat pore pattern was identified on the skin surface with a methylene blue stain driven in by iontophoresis (15). The entire surface of epithelium in an area containing approximately five sweat pores was "burred" off using a fine-grained conical-shaped abrasive driven by a highspeed rotary motor (Fig. 2). In a 2 cm. square area two or three such injuries were produced (Fig. 3). Bleeding was not encountered. The area studied was the intrascapular and upper lumbar areas of the back. Biopsies were takea in one series of experiments immediately after burring and 4, 8, 12, 24, and 48 hours and 3, 5,
7, 9, 12, and 14 days after burring. In a second series of experiments biopsies
were taken 3, 3, 4, 4j, and 5 days after burring. The dermis was burred clean of all epithelial cells (Fig. 4). A description of the
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FIG. 4. Site of injury four hours after burring (a) dermal surface free of epidermal cells (b) injured superficial portion of duct with pyknosis of most superficial of remaining cells. (Toluidine blue buffered pH 4.8). Mag. X 320.
morphological and histochemical responses of the epidermis in general to such
injnry will be reported at a later date. However, it can be said now that the epidermis at the edges of the injnry responded in much the same manner as was described by Loeb in 1898 (16). and by many others since then (17). This report will deal only with the responses of the eccrine sweat duct. The "Epidermal Sweat Duct Unit" was completely removed by the injury. Immediately after burring the distal end of the eccrine sweat duct developed degenerative changes of pyknosis and shriveling of its lumenal and basal cells to a depth of four or five cells; perhaps due to the heat or friction of this type of injury (Fig. 4). Glycogen disappeared from these cells but the other Schiffpositive material lining the lumen remained undisturbed. Nothing striking occurred for the first 24 hours except that a zone of necrobiosis began to develop in the exposed dermis approximately the thickness of the epidermis. This was sharply walled off from the underlying dermis with increasing numbers of in-
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RESPONSES OF HUMAN SWEAT DUCT
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fiammatory cells. In this degenerative zone the distal end of the sweat duct was trapped and also gradually underwent degenerative changes of pycnosis and shriveling of the cells with loss of glycogen. Its location was easily identified, however, by the persistence of the Schiff-positive material that previously lined the lumeiral cells. Between 24 and 48 hours after injury this remnant of duct
was pinched off from its viable end just under the necrobiotic area (Fig. 5). Regeneration of the duct was obviously under way three days after injury. (This response was some 24 hours behind similar responses in the marginal epidermis and in the pilosebaceous units). The first evidence of regeneration from
the duct was a marked swelling of the cytoplasm and nuclei of the basal and lumenal cells and the associated nuclear changes in the basal cells that are known to occur in the premitotic phase of cell activity. Since the duct is well encased in connective tissue, such swelling caused crowding of the cells into the lumen. Such crowding of the lumenal cells in turn, caused the lumen itself to become slightly tortuous (Fig. 6). More striking was the movement of the lumenal
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cells out of the open distal end of the duet. With no duet walls to contain them, they seemed to spill out in a cone-shaped manner under the necrobiotic dermis which was now obviously becoming a crust. In so doing, however, they spilled out in a tightly folded and spiraled tube that reminded one of ribbon candy that folds upon itself. (Figs. 6, 7) Since there was not enough mitotic activity to ex-
plain this early movement of these lumenal cells, one must explain it by the crowding due to increase in cell size and perhaps to a similar "gliding" of cells that is known to occur during repair of epidermal epithelium and other lining epithelia. At this time these lumenal cells of the duct are free of glycogen hut do have already the Schiff-positive material on their lumenal surfaces. Their cytoplasm
is not basophilic. The large basal cells of the duct are poor in Schiff-positive material but are accummulating more basophilic and metachromatic material in their cytoplasm. The basal cells have also spilled out of the end of the duct so that the entire structure three days after the injury resembles the silhouette
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Fin. 7. Four day response. (a) Protrusion of lumenal cells in form of tightly spiralled and coiled tube against overlying crust. (b) Piling up and spilling over of basal cells at margin of duct opening. (c) Mitotic acitivity in basal cells of duct. (d) A tongue of epithelium from the neclc of the funnel on the right. (Toluidine Blue stain, pH 5). Mag. X 600.
of a funnel (Fig. 6). It is from the neck of the funnel that the basal cells then begin to move. The response of regeneration and growth of the sweat duct becomes dramatic between three and five days after injury. Interestingly, all activity seems limited to the distal 20 or 30 cells of the dermal duct. Below this point in the mid-dermis the duct and its underlying secretory coil of the gland do not become involved in the problem of repair; histochemically and morph ologically they remain serene as normal resting structures. Three to three and one-half days after injury, the basal cells of the distal end of the duct abound in mitotic activity with the associated depletion of glycogen
and accummulation of metachromatic and basophilic material in their cytoplasms. These basal cells ow act as indifferent cells with two missions to achieve. One is to differentiate into lumenal cells and the other is to supply a source of epithelial cells for repair of the epidermis. They do this latter job at the neck of
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the funnel of regrowth (Fig. 7). Here mitosis and cell "gliding" are prominent for the next two days, so that these indifferent basal cells form a wall two or three cells thick about the duct and at the same time send out long tongues of at first one layer of swollen prickle cells under the inflammatory cells that demarcate the dermis from the crust. Immediately, the gliding prickle cells that are to be part of the general epidermis accummulate glycogen, while the other prickle cells about the funnel neck as well as the lumenal cells remain glycogen free. In rapid order then the tongues of epithelium become sheets of three or more cells in depth (Fig. 8). By the fifth day after injury the sheets of new epithelium from all sources have met to form an intact cover over the dermis and beneath the rapidly shrinking crust. During this time the lumenal cells are rapidly folding and spiralling their tube so that by four and one-half days after injury as many as six to nine twists can be counted above the neck of the funnel. At times this single cell-walled tube
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RESPONSES OF HUMAN SWEAT DUCT
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FIG. 9. Four and one-half day response (high power of Fig. 8). The tightly folded and coiled tube of lumenal cells protrudes from funnel of regrowth and extends to the right beneath the crust on a thin lamina of migrating epidermal cells. (Toluidine Blue stain, pH 5). Mag. X 320.
runs far out in a straight line over the top of the tongue of the new epithelium (Fig. 9). All of the lumenal Cells both within the duet and in the "funnel" develop heavy amounts of subnuclear metachromasia (not normally present) and although they contain no glycogen, the n3n-glycogen Schiff-positive material is prominent on the lumenal margins of these cells. (One individual Who is known to have the ability to develop miliaria rubra developed a few casts of this Schiffpositive material in the lumen of the duet at approximately the level of the neck of the "funnel".) The lumenal cells below the twisting and folding are columnar in shape with their distal cytoplasm of a ground glass appearance, at times protruding freely into the lumen. The lumeaal cells in the spiral are flat, elongated cells with metachromasia on either side of and below the nucleus. Just as the degeneration and inactivity of the first forty-eight hours after injury is replaced by the hurried cellular activity of the three to five days post injury response, so is this in turn replaced by a slower "settling-in process," re-
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organization and orientation so to speak, between the five and fourteen days after the injury. At five days after injury there are prickle cells surrounding the lumenal cells at the neck of the "funnel". These cells are four to six deep and are in turn surrounded by still mitoti.cally active basal cells. Such prickle cells are small, intensely basophilic in their cytoplasm and weak in glycogen. These will become "the periductal sheath cells" of the "Epidermal Sweat Duct Unit". They are easily distinguished from the large swollen prickle cells of the adjacent newly repaired epidermis which are rich in glycogen and poor in basophilism
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(Fig. 10). The tube of lumenal cells, whose open end has always remained free on top of the underlying new-formed epithelium, is now picked up and surrounded by the prickle cells. Between seven and nine days the crust has fallen off exposing a hyperkeratotic epidermis with a granular layer. Now the previously tightly folded coils and spirals of the sweat duct tube with
Fin. 10. Five day response. (a) Lumenal cells, basophilic and metachromatie. (b) Penductal sheath of small prickle cells concentrically arranged and heavy with basophilic and metachromatic material. (c) Adjacent area of epidermal cells with less metachromasia. (Toluidine Blue stain, pH 4.8). Mag. X 640.
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RESPONSES OF HUMAN SWEAT DUCT
its lumenal Cells lying directly on top of lumenal cells is stretched out with only three or four turns to the spiral. These thicker, hyalinized lumenal cells are now surrounded by two or three layers of concentrically arranged prickle cells; thus forming the periductal sheath of epidermal cells of the "unit".
In the new epidermis seven days after injury, the so-called keratohyalin granules are accumulating three cells deep under the stratum corneum in large prickle cells that are still very rich in glycogen. The lumenal cells of the sweat duct, though glycogen free, are also becoming laden with granules in the outer half of their epidermal course. Fourteen days after injury (Fig. 11), the new epidermis is hyperkeratotic. In the "Epidermal Sweat Duct Unit" the lumenal cells are glycogen free and those lying in the outer half of the epidermis contain keratohyalin granules. The nonglycogen Schiff-positive material on the surface of these cells is heavy and is
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continuous into the pore. The "periductal sheath" is now two to three pickle cells thick. These cells are smaller and poorer in glycogen than the adjacent prickle cells and thus are easily identified. The basal cells about the apex of this "unit" are smaller than the adjacent basal cells of the epidermis. They contain no mitotic activity and are glycogen free. The sweat pore is hyperkeratotic along with the epidermis in general. Its keratin, however, is denser and stains bluer with hematoxylin than the adjacent stratum corneum. Within this dense keratin ring is the ring of Schiff-positive (non-glycogen) material that lines the lumen
of the pore. Thus, that portion of the eccrine sweat duct that lies within the epidermis is once again intact as a mass of cells containing (1) a lumenal cell lining the duct (2) the epidermal sheath of periductal cells with their basal cells about the apex of the mass and (3) the sweat pore. DI5CU5SION
One way to study the growth and development of a structure is to selectively injure it and observe its responses following such an injury. Such an approach to the better understanding of the "Epidermal Sweat Duct Unit" was undertaken in this report. The "unit" was not sufficiently altered by the superficial injury of the strip method (Pinkus) to give us the information we desired. Attempts at specific injury to the unit by discrete currettments under high magnification was technically a failure for two reasons. [11 the unit was tougher and more difficult to scrape off than the adjacent epidermis and [2] in the curretting process cells are implanted topsy-turvy into the area as quickly as they are removed (more like turning soil over with a plow than scraping it away). A technic of "burring" off the entire epidermis with a motor driven highspeed rotary fine-grained conical shaped abrasive effectively removes the "unit" and the epidermis about it, thus allowing one to watch it regrow first as an isolated structure and then as an important part of the general epidermal repair. In brief there are three maj or stages to the repair of the eccrinc sweat duct after burring. [1] The stage of degeneration and the preparation of the environment for the regeneration to follow. This takes place during the first forty-eight hours after injury; [2] the stage of active cell migration and proliferation which occurs three to five days after injury; 3] the stage of reorganization and orientation of the "Epidermal Sweat Duct Unit" to the remainder of the regenerating epidermis. This occurs after five days following injury. In the stage of active cell migration and proliferation several points are of particular interest. [1] The lumenal (lining) cells of the duct migrate out into the dermis to produce abundant tightly folded spirals before any adjacent epidermis is present. They do this at first before any great mitotic activity is evident and then at a rapid rate with the increase of mitosis in the basal cells of the duct. [2] The basal cells of the duct act as indifferent epidermal cells which (a) can be differentiated into the lumenal cells as well as (b) send out sheets of new prickle cells to help lay down a new epidermis. [3] All of this active growth arises
from and takes place in only a small zone at the distal end of the sweat duct; that part of the duct that is to become the apex of the new "Epidermal Sweat Duct Unit". The remainder of the sweat duct and its associated secretory coil
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are not altered morphologically or histochemically (McManus procedure with saliva or Toluidine Blue with ribonuclease) from their normal state. Such observations seem to indicate that for this type of injury, at least, the growth center for the "Epidermal Sweat Duct Unit" lies in those basal cells of the sweat duct where the duct joins the epidermis. These basal cells possess the ability to differentiate into prickle cells (and thus the periductal sheath of epidermal cells) as well as into the highly specialized lumenal cells lining the duct. Studies to find wherein lies the differentiating determinants for the growth of these cells and structures are in progress. SUMMARY
The "Epidermal Eccrine Sweat Duct Unit" was selectively removed along with the adjacent epidermis bymeans of a high-speed rotary fine-grained abrasive.
Observations of the regeneration of the sweat duct indicate: (a) its growth center to be from a small zone of basal cells where the sweat duct joins the epidermis; (b) these basal cells are indifferent epidermal cells that can become either highly specialized lumenal cells lining the sweat duct or prickle cells; (c) the lumenal cells lining the "Epidermal Sweat Duct Unit" have the ability to produce spirals and coils in the sweat duct before any epidermis is present. REFERENCES C., Ja., HOLYOKE, J. B. AND MONTAGNA, W.: The "Epidermal Sweat Duet Unit", a morphologic and biologic entity. J. Invest. Dermat., 22: 157, 1954. 2. O'BRIEN, J. P.: The etiology of poral closure. J. Invest. Dermat., 15: 95, 1950.
1. LOBITZ, W.
3. BUNTING, H., WISL0cKI, G. B. AN!) DEMPSEY, E. W.: The chemical histology of human eccrinc and apocrine sweat glands. Anat. Rec., 100: 61, 1948. 4. MONTAGNA, W., CHASE, R. B. AND HAMILTON, J. B.: The distribution of glycogen and
lipids in human skin. J. Invest. Dermat., 17: 147, 1951. 5. MONTAGNA, W., CHASE, H. B. AND LOBITZ, W. C., Ja.: Histology and cytochemistry
of human skin. II. The distribution of glycogcn in the epidermis, hair follicles, scbaceous glands, and eccrine sweat glands. Anat. Rcc., 114: 231, 1952. 6. SHELLEY, W. B. AND MESCON, H.: Histochemical demonstration of secretory activity in human cccrine sweat glands. J. Invest. Dermat., 18: 289, 1952. 7. M0NTAiNA, W., CHASE, H. B. AND L0BITZ, W. C., Ja.: Histology and cytochemistry of human skin. IV. The cccrine sweat glands. J. Invest. Dcrmat., 20: 415, 1953.
8. HOEPKE, H.: (a) Die hant, in handbuch der mikrcskopischen anatomie des menschen (W.V. 1Ioellendorff)., III, 1:24, Berlin, 1927. (b) Dcr cpidcrmalc teil des ausfuehrungsganges der ekkrinen schweissdruesen. Ztschr. f. Anat. u. Entwcklngsgcsch., 87: 319, 1928.
9. PINKUS, H.: Notes on the anatomy and pathology of the skin appendages. I. The wall of the intra-epidermal part of the sweat duct. J. Invest. Dermat., 2: 175, 1939. 10. HOLYOKE, J. B. AND LOBITZ, W. C., JR.: Histologic variations in the structure of human
eccrine sweat glands. J. Invest. Dermat., 18: 147, 1952. 11. PINKaTS, H.: Examination of the epidermis by the strip method. II. Biometric data on regeneration of the human epidermis. J. Invest. Dcrmat., 19: 431, 1952. 12. URBAcH, F. AND SHELLEY, W. B.: A rapid and simple method for obtaining punch biopsies without anesthesia. J. Invest. Dermat., 17: 131, 1951. 13. McMANU5, J. F. A.: Histologic and histochemical uses of periodic acid. Stain Technol., 23: 99, 1948.
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14. L0BITz, W. C., JR. AND HOLYOKE, J. B.: The histochemical response of the human epidermis to controlled injury; glycogen. J. Invest. Dermat., 22: 189, 1954.
15. REIN, H.: Zur electrophysiologie der manschlichen haut: I. Untersuchungen uber farbstoffein wanderung in lebende warm bluter haut im electrisehen felde. Ztschr. f. Biol., 84: 41, 1926.
16. LOEB, L.: Ueber regeneration des epithels. Arch. 1. Entwcklngsmechn. d. Organ., 6: 297, 1898.
A comparative study of the mechanim of wound healing. J. Med. Research, 41: 247, 1920.
17. AREY, L. B.: Wound healing. Physiol. Rev., 16: 327, 1936.
DISCUSSION DR. STEPHEN ROTHMAN, Chicago, Iii.: The last slide shows the Malpighian
layer loaded with glycogen. It is known that this happens in regenerating epithelium after injury and in acanthotic epidermis in general. However, I wonder if the glycogen stays there permanently. I thought that it disappears after the repair process is completed or the epidermis becomes normal in all respects. How
long does it take for the glycogen to disappear? DR. RAYMOND R. SUSKIND, Cincinnati, Ohio: I would like to compliment Dr.
Lobitz on a very beautiful presentation. He has given me some ideas as to how to proceed to answer some of my own questions with respect to sweating in acanthotic skin diseases such as psoriasis in which sweat function is altered.
I would like to ask him if he has made any histochemical observations in cutaneous problems such as psoriasis or non-specific dermatoses in which there is proliferation of the epidermis. I am particularly interested in the reaction of the basal cells and of the cells lining the epidermal portion of the sweat duct. I would like to know if he has ever noted effacement of the duct lumen by cell proliferation and keratinization and then subsequent restitution. DR. WALTER C. L0BITz (in closing): I want to thank the discussors very much.
The accumulation of glycogen in the mid-epidermis seems to occur after the burst of mitotic activity in the basal cells and before complete healing in any injured skin. The onset of this accumulation varies with the degree of epidermal damage. It is usually present within 48 hours after the phase of mitotic activity. Whether such glycogen accumulation is there to furnish energy for keratinization or whether it's a damming up of glycogen due to the slowing down of cellular
activity is still to be established. The duration of this glycogen accumulation again varies with the degree and duration of the injury. If acanthosis is produced it will always be present. After a single, mild injury glycogen begins to disappear from these cells at approximately 14 days. In answer to Dr. Suskind: I think that Dr. Cormia has done a very nice pres-
entation recently at the A.D.A. on just such a problem, and Dr. Blank, who has worked with him, might be able to answer that more specifically than I could.
In our observations, the epidermal sweat duct unit does not seem to be involved; but we have not deliberately taken a unit in an abnormal pathological process like psoriasis and injured it and watched it regenerate. That should be done.
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94
linolenic acid extract. Arch. This pdf is a scanned copy UV of irradiated a printed document.
24. Wynn, C. H. and Iqbal, M.: Isolation of rat
skin lysosomes and a comparison with liver Path., 80: 91, 1965. and spleen lysosomes. Biochem. J., 98: lOP, 37. Nicolaides, N.: Lipids, membranes, and the 1966.
human epidermis, p. 511, The Epidermis
Eds., Montagna, W. and Lobitz, W. C. Acascopic localization of acid phosphatase in demic Press, New York. human epidermis. J. Invest. Derm., 46: 431, 38. Wills, E. D. and Wilkinson, A. E.: Release of 1966. enzymes from lysosomes by irradiation and 26. Rowden, C.: Ultrastructural studies of kerathe relation of lipid peroxide formation to tinized epithelia of the mouse. I. Combined enzyme release. Biochem. J., 99: 657, 1966. electron microscope and cytochemical study 39. Lane, N. I. and Novikoff, A. B.: Effects of of lysosomes in mouse epidermis and esoarginine deprivation, ultraviolet radiation and X-radiation on cultured KB cells. J. phageal epithelium. J. Invest. Derm., 49: 181, 25. Olson, R. L. and Nordquist, R. E.: Ultramicro-
No warranty is given about the accuracy of the copy.
Users should refer to the original published dermal cells. Nature, 216: 1031, 1967. version of1965. the material. vest. Derm., 45: 448, 28. Hall, J. H., Smith, J. G., Jr. and Burnett, S. 41. Daniels, F., Jr. and Johnson, B. E.: In prepa1967.
Cell Biol., 27: 603, 1965.
27. Prose, P. H., Sedlis, E. and Bigelow, M.: The 40. Fukuyama, K., Epstein, W. L. and Epstein, demonstration of lysosomes in the diseased J. H.: Effect of ultraviolet light on RNA skin of infants with infantile eczema. J. Inand protein synthesis in differentiated epi-
C.: The lysosome in contact dermatitis: A ration. histochemical study. J. Invest. Derm., 49: 42. Ito, M.: Histochemical investigations of Unna's oxygen and reduction areas by means of 590, 1967. 29. Pearse, A. C. E.: p. 882, Histochemistry Theoultraviolet irradiation, Studies on Melanin, retical and Applied, 2nd ed., Churchill, London, 1960.
30. Pearse, A. C. E.: p. 910, Histacheini.stry Thearetscal and Applied, 2nd ed., Churchill, London, 1960.
31. Daniels, F., Jr., Brophy, D. and Lobitz, W. C.: Histochemical responses of human skin fol-
lowing ultraviolet irradiation. J. Invest. Derm.,37: 351, 1961.
32. Bitensky, L.: The demonstration of lysosomes by the controlled temperature freezing section method. Quart. J. Micr. Sci., 103: 205, 1952.
33. Diengdoh, J. V.: The demonstration of lysosomes in mouse skin. Quart. J. Micr. Sci., 105: 73, 1964.
34. Jarret, A., Spearman, R. I. C. and Hardy, J. A.:
Tohoku, J. Exp. Med., 65: Supplement V, 10, 1957.
43. Bitcnsky, L.: Lysosomes in normal and pathological cells, pp. 362—375, Lysasames Eds., de Reuck, A. V. S. and Cameron, M. Churchill, London, 1953.
44. Janoff, A. and Zweifach, B. W.: Production of inflammatory changes in the microcirculation by cationic proteins extracted from lysosomes. J. Exp. Med., 120: 747, 1964.
45. Herion, J. C., Spitznagel, J. K., Walker, R. I. and Zeya, H. I.: Pyrogenicity of granulocyte lysosomes. Amer. J. Physiol., 211: 693, 1966.
46. Baden, H. P. and Pearlman, C.: The effect of ultraviolet light on protein and nucleic acid synthesis in the epidermis. J. Invest. Derm.,
Histochemistry of keratinization. Brit. J. 43: 71, 1964. Derm., 71: 277, 1959. 35. De Duve, C. and Wattiaux, R.: Functions of 47. Bullough, W. S. and Laurence, E. B.: Mitotic control by internal secretion: the role of lysosomes. Ann. Rev. Physiol., 28: 435, 1966. the chalone-adrenalin complex. Exp. Cell. 36. Waravdekar, V. S., Saclaw, L. D., Jones, W. A. and Kuhns, J. C.: Skin changes induced by
Res., 33: 176, 1964.
WALTER C. LOBITZ, JR., M.D., JOHN B. HOLYOKE, M.D. AND WILLIAM MONTAGNA, Pn.D.
The human ecerine sweat duct may be divided into two parts. [1] The "dermal
sweat duct" (that portion that lies within the dermis) and [2] the "epidermal sweat duct unit" (that portion that lies within the epidermis). The "dermal duct" is composed of two layers of cells. [1] The basal cells which rest on the connective tissue sheath are usually euboidal and contain occasional mitotic figures. [2] The "lumenal cells" which line the lumen of the sweat duet are euboidal or columnar, have a hyalinized lumenal border that varies in size, and are never seen to contain mitotie figures. The "Epidermal Sweat Duct Unit" (1) is composed of three major parts. [1] The intraepidermal portion of the sweat duet which is made up of the single layer of cells lining the lumen and is a continuation of the lumenal cells of the dermal duet. These lumenal or lining cells are euboidal or rectangularly flat when arranged about the lumen. They do not have priekies between themselves, they have a clear cytoplasm in the lo\ver two-thirds of the epidermis, but accumulate "Keratohyalin" granules in the outer one-third of the epidermis. [2] The periduetal sheath of two or more epidermal (prickle) cells that are concentrically arranged around the lumenal cells. These sheath cells form a mass of cells within the epidermis. About the apex of this cell mass are pigment-free basal cells that are continuous with the basal cells of the dermal duet on the one side, and with the basal cells of the adjacent epidermis on the other. The flat end of the cell mass, in the center of which is the sweat pore, is on the surface of the skin. [3] The sweat pore (the ecerine sweat duct unit opening on the surface of the skin)
is encircled by a ring of keratin fibers (O'Brien's Keratin Ring (2)) that are, adjacent to the keratin fibers of the stratum corneum. Within the keratin ring and lining the lumen of the pore, is a ring of Sehiff-positive material (not glyeogen) that contains less disulfide concentrations than the keratin ring and is continuous with a similar Sehiff -positive material that covers the free surface of all of the lumenal cells of the entire dermal and epidermal eeerine sweat duet. The histoehemieal contents of these various cells in the dermal and epidermal
portions of the human eeerine sweat duet have been described and add more * From the Department of Dermatology (W.C.L., Jr.) and the Department of Pathology (J.B.H.) Hitchcock Clinic and Dartmouth Medical School, Hanover, New Hampshire;
and the Arnold Biological Laboratory (W.M.) Brown University, Providence, Rhode Island.
This study was supported by the Quartermaster Research and Development Command (Contract ,DA44-1O9 q.m. 1424) and by the Hitchcock Foundation.
Presented at the fifteenth Annual Meeting of The Society for Investigative Dermatology, Inc., San Francisco, California, June 19, 1954. 329
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evidence to the similarities, differences, and "biological structure" of their component parts (1, 3—7).
Hoepke (8) was among the first to point out that the sweat duct in the epi-
dermis was a closed tube of lining cells and not merely a canal-like space bet\veen
prickle cells. However, "he insists that the wall is but a functional entity and that its building stones are the prickle cells of the epidermis." (9) Pinkus (9) reviewed this and added evidence to a concept that the epidermal portion of the sweat duct behaved independently and differently from the rest of the epidermis. Among other things he pointed out that in the epidermis: [1] The duct was un-
affected by the surrounding edema and vesiculation in experimental allergic dermatitis. [2] The duct retained its normal structure even \vhen surrounded by the dyskeratotic cells of senile keratoses and the anaplastic cells of Bowen's disease. [3] There are no melanin granules in the duct cells of normal Negro or hyperpigmented white skin, nor in the stratum corneum overlying the sweat duct. (We have never seen pigment in the basal cells at the apex of the cone of the "epidermal sweat duct unit" even though the adjacent basal cells are heavily laden with such pigment.) [4] The epidermal sweat duct spirals to accommodate its length to the adjacent epidermis; it is straight in the acanthotic epidermis of psoriasis and tightly coiled when the epidermis is narrowed with atrophy. Pinkus then raised the question as to where the "matrix for the replacement of these cells could be". He postulated, on the evidence then at hand, that the entire length of the sweat duct might serve as the matrix by mitotic cell division. He states that histopathologically "adenomatous proliferation of the syringeal
epithelium is not uncommonly found near lupus vulgaris, after roentgen-ray treatment of carcinomas and in other chronic processes which destroy the upper ends of the ducts"; and "it is a long recognized fact that remnants of the appendages help materially in the epithelialization of granulating wounds". Such types of questions are always pertinent to those interested in normal and abnormal growth patterns in the human. Questions concerning the ecerine sweat duct are of special interest to those focusing their attention on the structure and the function of the human eccrine sweat gland and on the sweat retention
group of diseases in particular. For such reasons, histologic studies (10) were made and confirmed some of Pinkus' observations. Recent histochemieal studies added more evidence that the "Epidermal Sweat Duct Unit" (1) is a biologic as well as a morphologic entity that behaves differently from the epidermis and as such must be further investigated. The following observations have resulted from some of our attempts to better understand this ecerine sweat duct in the human. PROCEDURE
This report will describe the effects of t\vo different types of injury to the epidermis and to the ecerine sweat duet on the growth responses of the sweat duet. The effect of such injury to the growth of the epidermis and to the pilosebaceous units will be described at a later date. The injuries to be reported in this paper are: [1] "The Strip Experiment":
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RESPONSES OF HUMAN SWEAT DUCT
Removal
of the stratum corneum of the epidermis by the Strip Method of
Pinkus (11) [2] "The Burr Experiment": The removal of the entire epidermis including the "Epidermal Sweat Duct Unit" with a high-speed rotary driven abrasive. Young adults, human males, were utilized in these studies. The areas injured were on the skin of the back in the sites of predilection for the sweat retention group of diseases and where the sweat gland population count was known to be high.
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All biopsies were taken without local anesthesia using the highspeed motor driven rotary 5 mm. punch (12). For this report the specimens were fixed immediately in chilled ilelley's solution. The tissue sections were stained with [1] hematoxylin and phloxine [2] Schiff reagent (13) with and without hematoxylin as a eounterstain and controlled with saliva for the removal of glycogen [3] Toluidine Blue, buffered at pH 5, and controlled with ribonuelease.
FIG. 1. The "Epidermal Sweat Duct Unit" immediately after stripping. The swollen basal cells of the adjacent epidermis sharply demarcate the unaltered "unit" (McManus procedure counterstained with hematoxylin) Mag. X 410.
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The Strip Experiment Seven young adult males were utilized. The area for study selected was the skin of the back between the lateral markings of the scapuli and posterior axillary
fold. A linear area of stratum corneum was stripped off with scotch tape. In order to get consistent stripping and response from subject to subject and during
repeated experiments in the same subject, the epidermis was stripped until moisture appeared on the skin. To accomplish this, for example, from 16—24 strippings were necessary on comparable areas of skin even in the same individual. Biopsies taken after stripping were as follows: [1] one group of men 16, 24, 48,
and 72 hours after stripping; [2] one group of three men 3, 5, 10, and 15 days after stripping; [3] one man immediately after stripping (approximately 5 minutes from the onset of injury), 4, 8, 16, 24, 48, and 72 hours and 5, 10, and 15 days after stripping.
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A most striking change occurred in the biopsy taken immediately after the injury. In the general epidermis the basal cells became swollen to almost t\vice the original diameter; the cytoplasm was clear and completely devoid of any stain or counterstain (14). The basal cells of the "Epidermal Sweat Duct Unit," on the other hand, did not undergo any such swelling (Fig. 1). Such swollen adjacent basal cells of the epidermis demarcate the "Epidermal Sweat Duct Unit" in even greater contrast. In addition the glycogen accumulation and utilization that occurred over the next 5 days after injury, did not take place in the respective
FIG. 2. The conical shaped fine abrasive used to burr off the epidermis.
RESPONSES OF HUMAN SWEAT DUCT
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cells of the "Epidermal Sweat Duet Unit" at any time, nor was there any burst of mitotie activity in these cells. The Burr Experiment
Three adult males were used. The sweat pore pattern was identified on the skin surface with a methylene blue stain driven in by iontophoresis (15). The entire surface of epithelium in an area containing approximately five sweat pores was "burred" off using a fine-grained conical-shaped abrasive driven by a highspeed rotary motor (Fig. 2). In a 2 cm. square area two or three such injuries were produced (Fig. 3). Bleeding was not encountered. The area studied was the intrascapular and upper lumbar areas of the back. Biopsies were takea in one series of experiments immediately after burring and 4, 8, 12, 24, and 48 hours and 3, 5,
7, 9, 12, and 14 days after burring. In a second series of experiments biopsies
were taken 3, 3, 4, 4j, and 5 days after burring. The dermis was burred clean of all epithelial cells (Fig. 4). A description of the
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FIG. 4. Site of injury four hours after burring (a) dermal surface free of epidermal cells (b) injured superficial portion of duct with pyknosis of most superficial of remaining cells. (Toluidine blue buffered pH 4.8). Mag. X 320.
morphological and histochemical responses of the epidermis in general to such
injnry will be reported at a later date. However, it can be said now that the epidermis at the edges of the injnry responded in much the same manner as was described by Loeb in 1898 (16). and by many others since then (17). This report will deal only with the responses of the eccrine sweat duct. The "Epidermal Sweat Duct Unit" was completely removed by the injury. Immediately after burring the distal end of the eccrine sweat duct developed degenerative changes of pyknosis and shriveling of its lumenal and basal cells to a depth of four or five cells; perhaps due to the heat or friction of this type of injury (Fig. 4). Glycogen disappeared from these cells but the other Schiffpositive material lining the lumen remained undisturbed. Nothing striking occurred for the first 24 hours except that a zone of necrobiosis began to develop in the exposed dermis approximately the thickness of the epidermis. This was sharply walled off from the underlying dermis with increasing numbers of in-
335
RESPONSES OF HUMAN SWEAT DUCT
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Fm. 5. Response twenty-four hours after injury. Development of sharply demarcated necrobiotic zone of superficial dermis that includes the superficial portion of the duct. (McManus procedure without counterstain). Mag. X 310.
fiammatory cells. In this degenerative zone the distal end of the sweat duct was trapped and also gradually underwent degenerative changes of pycnosis and shriveling of the cells with loss of glycogen. Its location was easily identified, however, by the persistence of the Schiff-positive material that previously lined the lumeiral cells. Between 24 and 48 hours after injury this remnant of duct
was pinched off from its viable end just under the necrobiotic area (Fig. 5). Regeneration of the duct was obviously under way three days after injury. (This response was some 24 hours behind similar responses in the marginal epidermis and in the pilosebaceous units). The first evidence of regeneration from
the duct was a marked swelling of the cytoplasm and nuclei of the basal and lumenal cells and the associated nuclear changes in the basal cells that are known to occur in the premitotic phase of cell activity. Since the duct is well encased in connective tissue, such swelling caused crowding of the cells into the lumen. Such crowding of the lumenal cells in turn, caused the lumen itself to become slightly tortuous (Fig. 6). More striking was the movement of the lumenal
336
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cells out of the open distal end of the duet. With no duet walls to contain them, they seemed to spill out in a cone-shaped manner under the necrobiotic dermis which was now obviously becoming a crust. In so doing, however, they spilled out in a tightly folded and spiraled tube that reminded one of ribbon candy that folds upon itself. (Figs. 6, 7) Since there was not enough mitotic activity to ex-
plain this early movement of these lumenal cells, one must explain it by the crowding due to increase in cell size and perhaps to a similar "gliding" of cells that is known to occur during repair of epidermal epithelium and other lining epithelia. At this time these lumenal cells of the duct are free of glycogen hut do have already the Schiff-positive material on their lumenal surfaces. Their cytoplasm
is not basophilic. The large basal cells of the duct are poor in Schiff-positive material but are accummulating more basophilic and metachromatic material in their cytoplasm. The basal cells have also spilled out of the end of the duct so that the entire structure three days after the injury resembles the silhouette
RESPONSES OF HUMAN SWEAT DUCT
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Fin. 7. Four day response. (a) Protrusion of lumenal cells in form of tightly spiralled and coiled tube against overlying crust. (b) Piling up and spilling over of basal cells at margin of duct opening. (c) Mitotic acitivity in basal cells of duct. (d) A tongue of epithelium from the neclc of the funnel on the right. (Toluidine Blue stain, pH 5). Mag. X 600.
of a funnel (Fig. 6). It is from the neck of the funnel that the basal cells then begin to move. The response of regeneration and growth of the sweat duct becomes dramatic between three and five days after injury. Interestingly, all activity seems limited to the distal 20 or 30 cells of the dermal duct. Below this point in the mid-dermis the duct and its underlying secretory coil of the gland do not become involved in the problem of repair; histochemically and morph ologically they remain serene as normal resting structures. Three to three and one-half days after injury, the basal cells of the distal end of the duct abound in mitotic activity with the associated depletion of glycogen
and accummulation of metachromatic and basophilic material in their cytoplasms. These basal cells ow act as indifferent cells with two missions to achieve. One is to differentiate into lumenal cells and the other is to supply a source of epithelial cells for repair of the epidermis. They do this latter job at the neck of
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the funnel of regrowth (Fig. 7). Here mitosis and cell "gliding" are prominent for the next two days, so that these indifferent basal cells form a wall two or three cells thick about the duct and at the same time send out long tongues of at first one layer of swollen prickle cells under the inflammatory cells that demarcate the dermis from the crust. Immediately, the gliding prickle cells that are to be part of the general epidermis accummulate glycogen, while the other prickle cells about the funnel neck as well as the lumenal cells remain glycogen free. In rapid order then the tongues of epithelium become sheets of three or more cells in depth (Fig. 8). By the fifth day after injury the sheets of new epithelium from all sources have met to form an intact cover over the dermis and beneath the rapidly shrinking crust. During this time the lumenal cells are rapidly folding and spiralling their tube so that by four and one-half days after injury as many as six to nine twists can be counted above the neck of the funnel. At times this single cell-walled tube
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Fin. 8. Four and one-half day response. (a) Tongue of epidermal cells advancing beneath crust to left of duct opening. (h) Protruding and coiled tube of lumerial cells kinked to right of duct. (Toluidine Blue stain, pH 5). Mag. X 145.
RESPONSES OF HUMAN SWEAT DUCT
339
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FIG. 9. Four and one-half day response (high power of Fig. 8). The tightly folded and coiled tube of lumenal cells protrudes from funnel of regrowth and extends to the right beneath the crust on a thin lamina of migrating epidermal cells. (Toluidine Blue stain, pH 5). Mag. X 320.
runs far out in a straight line over the top of the tongue of the new epithelium (Fig. 9). All of the lumenal Cells both within the duet and in the "funnel" develop heavy amounts of subnuclear metachromasia (not normally present) and although they contain no glycogen, the n3n-glycogen Schiff-positive material is prominent on the lumenal margins of these cells. (One individual Who is known to have the ability to develop miliaria rubra developed a few casts of this Schiffpositive material in the lumen of the duet at approximately the level of the neck of the "funnel".) The lumenal cells below the twisting and folding are columnar in shape with their distal cytoplasm of a ground glass appearance, at times protruding freely into the lumen. The lumeaal cells in the spiral are flat, elongated cells with metachromasia on either side of and below the nucleus. Just as the degeneration and inactivity of the first forty-eight hours after injury is replaced by the hurried cellular activity of the three to five days post injury response, so is this in turn replaced by a slower "settling-in process," re-
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organization and orientation so to speak, between the five and fourteen days after the injury. At five days after injury there are prickle cells surrounding the lumenal cells at the neck of the "funnel". These cells are four to six deep and are in turn surrounded by still mitoti.cally active basal cells. Such prickle cells are small, intensely basophilic in their cytoplasm and weak in glycogen. These will become "the periductal sheath cells" of the "Epidermal Sweat Duct Unit". They are easily distinguished from the large swollen prickle cells of the adjacent newly repaired epidermis which are rich in glycogen and poor in basophilism
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(Fig. 10). The tube of lumenal cells, whose open end has always remained free on top of the underlying new-formed epithelium, is now picked up and surrounded by the prickle cells. Between seven and nine days the crust has fallen off exposing a hyperkeratotic epidermis with a granular layer. Now the previously tightly folded coils and spirals of the sweat duct tube with
Fin. 10. Five day response. (a) Lumenal cells, basophilic and metachromatie. (b) Penductal sheath of small prickle cells concentrically arranged and heavy with basophilic and metachromatic material. (c) Adjacent area of epidermal cells with less metachromasia. (Toluidine Blue stain, pH 4.8). Mag. X 640.
341
RESPONSES OF HUMAN SWEAT DUCT
its lumenal Cells lying directly on top of lumenal cells is stretched out with only three or four turns to the spiral. These thicker, hyalinized lumenal cells are now surrounded by two or three layers of concentrically arranged prickle cells; thus forming the periductal sheath of epidermal cells of the "unit".
In the new epidermis seven days after injury, the so-called keratohyalin granules are accumulating three cells deep under the stratum corneum in large prickle cells that are still very rich in glycogen. The lumenal cells of the sweat duct, though glycogen free, are also becoming laden with granules in the outer half of their epidermal course. Fourteen days after injury (Fig. 11), the new epidermis is hyperkeratotic. In the "Epidermal Sweat Duct Unit" the lumenal cells are glycogen free and those lying in the outer half of the epidermis contain keratohyalin granules. The nonglycogen Schiff-positive material on the surface of these cells is heavy and is
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342
THE JOURNAL OF TNVE5TIGATTVE DERMATOLOGY
continuous into the pore. The "periductal sheath" is now two to three pickle cells thick. These cells are smaller and poorer in glycogen than the adjacent prickle cells and thus are easily identified. The basal cells about the apex of this "unit" are smaller than the adjacent basal cells of the epidermis. They contain no mitotic activity and are glycogen free. The sweat pore is hyperkeratotic along with the epidermis in general. Its keratin, however, is denser and stains bluer with hematoxylin than the adjacent stratum corneum. Within this dense keratin ring is the ring of Schiff-positive (non-glycogen) material that lines the lumen
of the pore. Thus, that portion of the eccrine sweat duct that lies within the epidermis is once again intact as a mass of cells containing (1) a lumenal cell lining the duct (2) the epidermal sheath of periductal cells with their basal cells about the apex of the mass and (3) the sweat pore. DI5CU5SION
One way to study the growth and development of a structure is to selectively injure it and observe its responses following such an injury. Such an approach to the better understanding of the "Epidermal Sweat Duct Unit" was undertaken in this report. The "unit" was not sufficiently altered by the superficial injury of the strip method (Pinkus) to give us the information we desired. Attempts at specific injury to the unit by discrete currettments under high magnification was technically a failure for two reasons. [11 the unit was tougher and more difficult to scrape off than the adjacent epidermis and [2] in the curretting process cells are implanted topsy-turvy into the area as quickly as they are removed (more like turning soil over with a plow than scraping it away). A technic of "burring" off the entire epidermis with a motor driven highspeed rotary fine-grained conical shaped abrasive effectively removes the "unit" and the epidermis about it, thus allowing one to watch it regrow first as an isolated structure and then as an important part of the general epidermal repair. In brief there are three maj or stages to the repair of the eccrinc sweat duct after burring. [1] The stage of degeneration and the preparation of the environment for the regeneration to follow. This takes place during the first forty-eight hours after injury; [2] the stage of active cell migration and proliferation which occurs three to five days after injury; 3] the stage of reorganization and orientation of the "Epidermal Sweat Duct Unit" to the remainder of the regenerating epidermis. This occurs after five days following injury. In the stage of active cell migration and proliferation several points are of particular interest. [1] The lumenal (lining) cells of the duct migrate out into the dermis to produce abundant tightly folded spirals before any adjacent epidermis is present. They do this at first before any great mitotic activity is evident and then at a rapid rate with the increase of mitosis in the basal cells of the duct. [2] The basal cells of the duct act as indifferent epidermal cells which (a) can be differentiated into the lumenal cells as well as (b) send out sheets of new prickle cells to help lay down a new epidermis. [3] All of this active growth arises
from and takes place in only a small zone at the distal end of the sweat duct; that part of the duct that is to become the apex of the new "Epidermal Sweat Duct Unit". The remainder of the sweat duct and its associated secretory coil
RESPONSES OF HUMAN SWEAT DUCT
343
are not altered morphologically or histochemically (McManus procedure with saliva or Toluidine Blue with ribonuclease) from their normal state. Such observations seem to indicate that for this type of injury, at least, the growth center for the "Epidermal Sweat Duct Unit" lies in those basal cells of the sweat duct where the duct joins the epidermis. These basal cells possess the ability to differentiate into prickle cells (and thus the periductal sheath of epidermal cells) as well as into the highly specialized lumenal cells lining the duct. Studies to find wherein lies the differentiating determinants for the growth of these cells and structures are in progress. SUMMARY
The "Epidermal Eccrine Sweat Duct Unit" was selectively removed along with the adjacent epidermis bymeans of a high-speed rotary fine-grained abrasive.
Observations of the regeneration of the sweat duct indicate: (a) its growth center to be from a small zone of basal cells where the sweat duct joins the epidermis; (b) these basal cells are indifferent epidermal cells that can become either highly specialized lumenal cells lining the sweat duct or prickle cells; (c) the lumenal cells lining the "Epidermal Sweat Duct Unit" have the ability to produce spirals and coils in the sweat duct before any epidermis is present. REFERENCES C., Ja., HOLYOKE, J. B. AND MONTAGNA, W.: The "Epidermal Sweat Duet Unit", a morphologic and biologic entity. J. Invest. Dermat., 22: 157, 1954. 2. O'BRIEN, J. P.: The etiology of poral closure. J. Invest. Dermat., 15: 95, 1950.
1. LOBITZ, W.
3. BUNTING, H., WISL0cKI, G. B. AN!) DEMPSEY, E. W.: The chemical histology of human eccrinc and apocrine sweat glands. Anat. Rec., 100: 61, 1948. 4. MONTAGNA, W., CHASE, R. B. AND HAMILTON, J. B.: The distribution of glycogen and
lipids in human skin. J. Invest. Dermat., 17: 147, 1951. 5. MONTAGNA, W., CHASE, H. B. AND LOBITZ, W. C., Ja.: Histology and cytochemistry
of human skin. II. The distribution of glycogcn in the epidermis, hair follicles, scbaceous glands, and eccrine sweat glands. Anat. Rcc., 114: 231, 1952. 6. SHELLEY, W. B. AND MESCON, H.: Histochemical demonstration of secretory activity in human cccrine sweat glands. J. Invest. Dermat., 18: 289, 1952. 7. M0NTAiNA, W., CHASE, H. B. AND L0BITZ, W. C., Ja.: Histology and cytochemistry of human skin. IV. The cccrine sweat glands. J. Invest. Dcrmat., 20: 415, 1953.
8. HOEPKE, H.: (a) Die hant, in handbuch der mikrcskopischen anatomie des menschen (W.V. 1Ioellendorff)., III, 1:24, Berlin, 1927. (b) Dcr cpidcrmalc teil des ausfuehrungsganges der ekkrinen schweissdruesen. Ztschr. f. Anat. u. Entwcklngsgcsch., 87: 319, 1928.
9. PINKUS, H.: Notes on the anatomy and pathology of the skin appendages. I. The wall of the intra-epidermal part of the sweat duct. J. Invest. Dermat., 2: 175, 1939. 10. HOLYOKE, J. B. AND LOBITZ, W. C., JR.: Histologic variations in the structure of human
eccrine sweat glands. J. Invest. Dermat., 18: 147, 1952. 11. PINKaTS, H.: Examination of the epidermis by the strip method. II. Biometric data on regeneration of the human epidermis. J. Invest. Dcrmat., 19: 431, 1952. 12. URBAcH, F. AND SHELLEY, W. B.: A rapid and simple method for obtaining punch biopsies without anesthesia. J. Invest. Dermat., 17: 131, 1951. 13. McMANU5, J. F. A.: Histologic and histochemical uses of periodic acid. Stain Technol., 23: 99, 1948.
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14. L0BITz, W. C., JR. AND HOLYOKE, J. B.: The histochemical response of the human epidermis to controlled injury; glycogen. J. Invest. Dermat., 22: 189, 1954.
15. REIN, H.: Zur electrophysiologie der manschlichen haut: I. Untersuchungen uber farbstoffein wanderung in lebende warm bluter haut im electrisehen felde. Ztschr. f. Biol., 84: 41, 1926.
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A comparative study of the mechanim of wound healing. J. Med. Research, 41: 247, 1920.
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DISCUSSION DR. STEPHEN ROTHMAN, Chicago, Iii.: The last slide shows the Malpighian
layer loaded with glycogen. It is known that this happens in regenerating epithelium after injury and in acanthotic epidermis in general. However, I wonder if the glycogen stays there permanently. I thought that it disappears after the repair process is completed or the epidermis becomes normal in all respects. How
long does it take for the glycogen to disappear? DR. RAYMOND R. SUSKIND, Cincinnati, Ohio: I would like to compliment Dr.
Lobitz on a very beautiful presentation. He has given me some ideas as to how to proceed to answer some of my own questions with respect to sweating in acanthotic skin diseases such as psoriasis in which sweat function is altered.
I would like to ask him if he has made any histochemical observations in cutaneous problems such as psoriasis or non-specific dermatoses in which there is proliferation of the epidermis. I am particularly interested in the reaction of the basal cells and of the cells lining the epidermal portion of the sweat duct. I would like to know if he has ever noted effacement of the duct lumen by cell proliferation and keratinization and then subsequent restitution. DR. WALTER C. L0BITz (in closing): I want to thank the discussors very much.
The accumulation of glycogen in the mid-epidermis seems to occur after the burst of mitotic activity in the basal cells and before complete healing in any injured skin. The onset of this accumulation varies with the degree of epidermal damage. It is usually present within 48 hours after the phase of mitotic activity. Whether such glycogen accumulation is there to furnish energy for keratinization or whether it's a damming up of glycogen due to the slowing down of cellular
activity is still to be established. The duration of this glycogen accumulation again varies with the degree and duration of the injury. If acanthosis is produced it will always be present. After a single, mild injury glycogen begins to disappear from these cells at approximately 14 days. In answer to Dr. Suskind: I think that Dr. Cormia has done a very nice pres-
entation recently at the A.D.A. on just such a problem, and Dr. Blank, who has worked with him, might be able to answer that more specifically than I could.
In our observations, the epidermal sweat duct unit does not seem to be involved; but we have not deliberately taken a unit in an abnormal pathological process like psoriasis and injured it and watched it regenerate. That should be done.
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94
linolenic acid extract. Arch. This pdf is a scanned copy UV of irradiated a printed document.
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C.: The lysosome in contact dermatitis: A ration. histochemical study. J. Invest. Derm., 49: 42. Ito, M.: Histochemical investigations of Unna's oxygen and reduction areas by means of 590, 1967. 29. Pearse, A. C. E.: p. 882, Histochemistry Theoultraviolet irradiation, Studies on Melanin, retical and Applied, 2nd ed., Churchill, London, 1960.
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