Int. 1. Exp. Path. (1990) 71, 713-7I6
Experimental frost-bite in Hanford Miniature Swine. III Sweat gland changes Polly Schoning Department of Pathology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas 665o6, USA
Received 28 April I 989 Accepted May I 990 ii
Summary. Frost-bite was produced in five Hanford Miniature Swine by exposure to 75C air for I, 3, 5, I0 or 20 min. Biopsies were taken at o, 3, 6, 2, 24 and 48 h and I, 2 and I 5 weeks. Sweat glands were evaluated microscopically and graded: o, no change to 5, severe change. Sweat gland changes were mild by h and moderate by 24 h for all freeze groups, except the min freeze group. Severe morphological changes were of two types: degeneration/necrosis and squamous metaplasia. These changes suggest that hyperhidrosis, as a sequel to frost-bite, may be more subjective than real and that squamous cell carcinoma, as a delayed sequel to frostbite, could originate from sweat glands as well as from the epidermis. -
1
i
i-
Keywords: frost-bite, hyperhidrosis, skin neoplasms
Hyperhidrosis has been reported as both a predisposing factor (Ervasti I962; Miller & Bjornson I962; Shafer & Thompson 1955) and as a sequel to frost-bite (Ervasti I962; Hermann et al. I 9 6 3; Lapp & Juergens I 9 6 5; Shafer & Thompson 1955). As a predisposing factor, hyperhidrosis has been linked to feelings of anxiety and the influence of epinephrine on the sweat glands. As a sequel to frost-bite, hyperhidrosis has been reported as common in as many as 40-63% of cases (Ervasti I962). Squamous cell carcinoma is a rare, but obviously severe, sequel to frost-bite (DiPirro & Conway I966; Katsas et al. 1977; Rossis et al. I982). In some instances, carcinoma has developed after as long as 20-2 5 years at the site of previous frost-bite. Although the pathogenesis of this neoplasm is not known, factors such as sunlight, industrial carcinogens (tars and oils), chronic ulcers, and old
burn scars have been implicated (Cotran et al. I989).
Reports of histopathologic changes of sweat gland injury with frost-bite are uncommon (Adams-Ray & Falconer I951; Dry 1946; Friedman 1946; Lapp & Juergens I 96 5). Our report documents the changes of sweat glands in pig skin exposed to - 75°C air for I, 3, 5, io and 20 min.
Materials and methods Experimental pigs and procedures have been described (Schoning & Hamlet I989). Briefly, frost-bite was produced using cold air (- 75sC) delivered to i-inch circular areas of skin on Hanford Miniature Swine. Skin was frozen for I, 3, 5, I0 or 20 min. Biopsies were taken at 0, 3, 6, 12, 24 and 48 h and at iand 2-week intervals. Tissues were fixed in io% buffered neutral formalin, then 7I3
7I4
P. Schoning
trimmed, embedded, sectioned, and stained with haematoxylin and eosin (H&E). Two hundred slides were evaluated using the following scale: o, no change; i, mild change; 2, mild-moderate; 3, moderate; 4, moderate-severe; 5, severe change. In addition, as part of another experiment, io biopsies were taken from a pig whose skin had been frozen 15 weeks previously. Results Sweat glands from unfrozen skin were arranged in packets of 20-2 5, surrounded by fat, and located in the deep reticular dermis. Each gland was lined by cuboidal epithelium and had a large lumen.
Sweat gland changes were mild by I h and became moderate by 24 h for all freeze groups, except the i- min freeze group for which changes remained mild. Earliest changes in the sweat glands were dilatation accompanied by flattened epithelium (Fig. i) or dilatation accompanied by lumens filled with sloughed epithelial cells (Fig. 2). More severe, later changes were of two types. The first type was characterized by glandular epithelial degeneration (Fig. 3), followed by necrosis, until finally only the outlines of glands remained (Fig. 4). The second type was characterized by hyperplasia of epithelial cells lining glands, in which the cells were piled up and the lumens obliterated (Figs 5 and 6). Eventually, those glands were
Fig. i. Dilated sweat glands lined by low cuboidal epithelium.
Fig. 3. Sweat glands with advanced epithelial cell degeneration. Lumens contain sloughed cells.
Fig. 2. Dilated sweat glands containing sloughed epithelial cells.
Fig. 4. Sweat glands with necrosis of epithelial cells. Cellular detail is lost.
Sweat gland injury
replaced by a solid sheet of squamous cells with individually keratinized cells interspersed (Figs 7 and 8). Sweat glands from the I1-week biopsies were characterized in two ways. The most common change was an absence of sweat glands. The other change was characterized by sweat glands that were decreased in number but essentially normal.
'W'
715
3 V
i
Fig. 7. Squamous metaplasia of sweat gland
h'.
^epithelium adjacent to a hair follicle.
-4~~~~~~~~~~~~~~~~~~~~~4
4f ~ Fig. 5. Epithelial hyperplasia of sweat glands adjacent to a hair follicle and surrounded by fat.
~
~
f
Nr, Fig. 8. Squamous metaplasia of sweat gland epithelium with cells arranged in a sheet. Indivi-
dually keratinized cells are present. Discussion
Hyperhidrosis is one of the commonest find_ings reported for frost-bite. However, the part it plays in frost-bite is unknown; reports e vary. It has been described as both a symptom and a sign (Shafer & Thompson I955). Psychological influences have been identi'&, / # fied in hyperhidrosis, along with anatomical ?. changes of sweat glands (Dry I946). Post> injury sweating has been observed to correFig. 6. Epithelial hyperplasia with cells growing late with pre-injury sweating (Ervasti I 962). into the lumens. Similarly, 4-h sweat rates of frost-bitten
^^
X
P. Schoning patients did not differ from those of controls Murray Hamlet, George Silver, Audrey Armstrong and Sgts Rodrigo Mariano, Mark (Lapp & Juergens I965). If hyperhidrosis is a sequel to frostbite, one Sharp, and George Staruch for their techwould expect to find morphologically normal nical help. Supported by US Army Faculty active sweat glands. However, we found that Research and Engineering Program (contract 86340) and Kansas Agricultural Expermost of the sweat glands were degenerating, necrotic, metaplastic, or absent in biopsy iment Station (contribution 89-42o-J). The samples ranging from o h to I 5 weeks. Others views, opinions, and/or findings contained in also have described microscopic changes of this report are those of the author and should sweat glands as shrivelled (Adams-Ray & not be construed as an official Department of Falconer I95I), hyalinized (Dry 1946), the Army position, policy, or decision, unless metaplastic (Friedman I946), or absent designated by other documentation. (Lapp & Juergens I965). Based on their reports and our observations, hyperhidrosis References would not be a likely sequel to frost-bite. Squamous cell carcinoma has occurred in ADAMS-RAY J. & FALCONER B. (I 95 I) Pathologicoanatomical changes following rapid and slow old frost-bite sites (DiPirro & Conway I966; thawing, respectively, in frozen skin in man. Katsas et al. I977; Rossis et al. I982). In our Acta Chir. Scand. 101, 269-27I. study, squamous metaplasia of sweat glands COTRAN, R.S., KUMAR V. & ROBBINS, S.L. (I989) was the most severe and dramatic change Robbins Pathologic Basis of Disease, Fourth edi(Figs 7 and 8). This change was seen most tion. Philadelphia: W.B. Saunders. pp. 34, 1287. often with the longer freezing times (5, I0 DIPIRRO E. & CONWAY H. (i 966) Carcinoma after and 20 min) and at the later biopsy times (i frostbite: A case report. Plast. Reconstr. Surg. and 2 weeks). Squamous metaplasia is known to be reversible, if the cause is DRY38,541-543. T.J. (I946) Experiences with late trench foot removed. It has been suggested that if some and frost-bite. Nebr. State Med. 31, 443-446. of the causes of metaplasia persist, cancer ERVASTI E. (I962) Frostbites ofthe extremities and their sequelae. Acta Chir. Scand. 299 (SuppI.), transformation might occur in that metai-69. plastic epithelium (Cotran et al. I989). FRIEDMAN N.B. (I946) The reactions of tissue to In conclusion, squamous metaplasia and cold. Am. Path. i6, 634-639. necrosis were the most severe changes seen HERMANN G.,1. Clin. SCHECHTER D.C., OWENS J.C. & in the sweat glands. These anatomical findSTARZL T.E. (I963) The problem of frostbite in ings suggest two concepts. Firstly, the sweat civilian medical practice. Surg. Clin. N. Am. 43, glands observed in this study could not have 5 I9-536. produced sweat. Therefore, hyperhidrosis as KATSAS A., AGNANTIS J., SMYRNIS S. & KAKAVOULIS T. (I977) Carcinoma on old frostbites. Am. J. a sequel to frost-bite may be more subjective Surg. 133, 377-378. than real. Secondly, the squamous metaLAPP N.L. & JUERGENS J.L. (I965) Frostbite. Mayo plasia seen in these sweat glands visually Clin. Proc. 40, 932-948. resembles a pre-neoplastic change. This MILLER D. & BJORNSON D.R. (i 962) An investiobservation suggests that frost-bite may be gation of cold injured soldiers in Alaska. Milit. one of the unproven influences of squamous Med. 127,247-252. cell carcinoma and that the site of origin RossIs C.G., YIACOUMETTIS A.M. & ELEMENOGLOU J. (I982) Squamous cell carcinoma of the heel could be sweat gland epithelium as well as at site of previous frostbite. I.R. Soc. developing the epidermis. Med. 75, 715-7I8.
7I6
Acknowledgements This experiment was done at USARIEM, Natick, MA, USA. The author thanks Drs
SCHONING P. & HAMLET M.P. (I989) Experimental frost-bite in Hanford Miniature Swine I Epithelial changes. Br. J. Exp. Path. 70, 41-49. SHAFER J.C. & THOMPSON A.C. (I955) Local cold injury. Arch. Dermatol. 72, 335-347.
Experimental frost-bite in Hanford Miniature Swine. III Sweat gland changes Polly Schoning Department of Pathology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas 665o6, USA
Received 28 April I 989 Accepted May I 990 ii
Summary. Frost-bite was produced in five Hanford Miniature Swine by exposure to 75C air for I, 3, 5, I0 or 20 min. Biopsies were taken at o, 3, 6, 2, 24 and 48 h and I, 2 and I 5 weeks. Sweat glands were evaluated microscopically and graded: o, no change to 5, severe change. Sweat gland changes were mild by h and moderate by 24 h for all freeze groups, except the min freeze group. Severe morphological changes were of two types: degeneration/necrosis and squamous metaplasia. These changes suggest that hyperhidrosis, as a sequel to frost-bite, may be more subjective than real and that squamous cell carcinoma, as a delayed sequel to frostbite, could originate from sweat glands as well as from the epidermis. -
1
i
i-
Keywords: frost-bite, hyperhidrosis, skin neoplasms
Hyperhidrosis has been reported as both a predisposing factor (Ervasti I962; Miller & Bjornson I962; Shafer & Thompson 1955) and as a sequel to frost-bite (Ervasti I962; Hermann et al. I 9 6 3; Lapp & Juergens I 9 6 5; Shafer & Thompson 1955). As a predisposing factor, hyperhidrosis has been linked to feelings of anxiety and the influence of epinephrine on the sweat glands. As a sequel to frost-bite, hyperhidrosis has been reported as common in as many as 40-63% of cases (Ervasti I962). Squamous cell carcinoma is a rare, but obviously severe, sequel to frost-bite (DiPirro & Conway I966; Katsas et al. 1977; Rossis et al. I982). In some instances, carcinoma has developed after as long as 20-2 5 years at the site of previous frost-bite. Although the pathogenesis of this neoplasm is not known, factors such as sunlight, industrial carcinogens (tars and oils), chronic ulcers, and old
burn scars have been implicated (Cotran et al. I989).
Reports of histopathologic changes of sweat gland injury with frost-bite are uncommon (Adams-Ray & Falconer I951; Dry 1946; Friedman 1946; Lapp & Juergens I 96 5). Our report documents the changes of sweat glands in pig skin exposed to - 75°C air for I, 3, 5, io and 20 min.
Materials and methods Experimental pigs and procedures have been described (Schoning & Hamlet I989). Briefly, frost-bite was produced using cold air (- 75sC) delivered to i-inch circular areas of skin on Hanford Miniature Swine. Skin was frozen for I, 3, 5, I0 or 20 min. Biopsies were taken at 0, 3, 6, 12, 24 and 48 h and at iand 2-week intervals. Tissues were fixed in io% buffered neutral formalin, then 7I3
7I4
P. Schoning
trimmed, embedded, sectioned, and stained with haematoxylin and eosin (H&E). Two hundred slides were evaluated using the following scale: o, no change; i, mild change; 2, mild-moderate; 3, moderate; 4, moderate-severe; 5, severe change. In addition, as part of another experiment, io biopsies were taken from a pig whose skin had been frozen 15 weeks previously. Results Sweat glands from unfrozen skin were arranged in packets of 20-2 5, surrounded by fat, and located in the deep reticular dermis. Each gland was lined by cuboidal epithelium and had a large lumen.
Sweat gland changes were mild by I h and became moderate by 24 h for all freeze groups, except the i- min freeze group for which changes remained mild. Earliest changes in the sweat glands were dilatation accompanied by flattened epithelium (Fig. i) or dilatation accompanied by lumens filled with sloughed epithelial cells (Fig. 2). More severe, later changes were of two types. The first type was characterized by glandular epithelial degeneration (Fig. 3), followed by necrosis, until finally only the outlines of glands remained (Fig. 4). The second type was characterized by hyperplasia of epithelial cells lining glands, in which the cells were piled up and the lumens obliterated (Figs 5 and 6). Eventually, those glands were
Fig. i. Dilated sweat glands lined by low cuboidal epithelium.
Fig. 3. Sweat glands with advanced epithelial cell degeneration. Lumens contain sloughed cells.
Fig. 2. Dilated sweat glands containing sloughed epithelial cells.
Fig. 4. Sweat glands with necrosis of epithelial cells. Cellular detail is lost.
Sweat gland injury
replaced by a solid sheet of squamous cells with individually keratinized cells interspersed (Figs 7 and 8). Sweat glands from the I1-week biopsies were characterized in two ways. The most common change was an absence of sweat glands. The other change was characterized by sweat glands that were decreased in number but essentially normal.
'W'
715
3 V
i
Fig. 7. Squamous metaplasia of sweat gland
h'.
^epithelium adjacent to a hair follicle.
-4~~~~~~~~~~~~~~~~~~~~~4
4f ~ Fig. 5. Epithelial hyperplasia of sweat glands adjacent to a hair follicle and surrounded by fat.
~
~
f
Nr, Fig. 8. Squamous metaplasia of sweat gland epithelium with cells arranged in a sheet. Indivi-
dually keratinized cells are present. Discussion
Hyperhidrosis is one of the commonest find_ings reported for frost-bite. However, the part it plays in frost-bite is unknown; reports e vary. It has been described as both a symptom and a sign (Shafer & Thompson I955). Psychological influences have been identi'&, / # fied in hyperhidrosis, along with anatomical ?. changes of sweat glands (Dry I946). Post> injury sweating has been observed to correFig. 6. Epithelial hyperplasia with cells growing late with pre-injury sweating (Ervasti I 962). into the lumens. Similarly, 4-h sweat rates of frost-bitten
^^
X
P. Schoning patients did not differ from those of controls Murray Hamlet, George Silver, Audrey Armstrong and Sgts Rodrigo Mariano, Mark (Lapp & Juergens I965). If hyperhidrosis is a sequel to frostbite, one Sharp, and George Staruch for their techwould expect to find morphologically normal nical help. Supported by US Army Faculty active sweat glands. However, we found that Research and Engineering Program (contract 86340) and Kansas Agricultural Expermost of the sweat glands were degenerating, necrotic, metaplastic, or absent in biopsy iment Station (contribution 89-42o-J). The samples ranging from o h to I 5 weeks. Others views, opinions, and/or findings contained in also have described microscopic changes of this report are those of the author and should sweat glands as shrivelled (Adams-Ray & not be construed as an official Department of Falconer I95I), hyalinized (Dry 1946), the Army position, policy, or decision, unless metaplastic (Friedman I946), or absent designated by other documentation. (Lapp & Juergens I965). Based on their reports and our observations, hyperhidrosis References would not be a likely sequel to frost-bite. Squamous cell carcinoma has occurred in ADAMS-RAY J. & FALCONER B. (I 95 I) Pathologicoanatomical changes following rapid and slow old frost-bite sites (DiPirro & Conway I966; thawing, respectively, in frozen skin in man. Katsas et al. I977; Rossis et al. I982). In our Acta Chir. Scand. 101, 269-27I. study, squamous metaplasia of sweat glands COTRAN, R.S., KUMAR V. & ROBBINS, S.L. (I989) was the most severe and dramatic change Robbins Pathologic Basis of Disease, Fourth edi(Figs 7 and 8). This change was seen most tion. Philadelphia: W.B. Saunders. pp. 34, 1287. often with the longer freezing times (5, I0 DIPIRRO E. & CONWAY H. (i 966) Carcinoma after and 20 min) and at the later biopsy times (i frostbite: A case report. Plast. Reconstr. Surg. and 2 weeks). Squamous metaplasia is known to be reversible, if the cause is DRY38,541-543. T.J. (I946) Experiences with late trench foot removed. It has been suggested that if some and frost-bite. Nebr. State Med. 31, 443-446. of the causes of metaplasia persist, cancer ERVASTI E. (I962) Frostbites ofthe extremities and their sequelae. Acta Chir. Scand. 299 (SuppI.), transformation might occur in that metai-69. plastic epithelium (Cotran et al. I989). FRIEDMAN N.B. (I946) The reactions of tissue to In conclusion, squamous metaplasia and cold. Am. Path. i6, 634-639. necrosis were the most severe changes seen HERMANN G.,1. Clin. SCHECHTER D.C., OWENS J.C. & in the sweat glands. These anatomical findSTARZL T.E. (I963) The problem of frostbite in ings suggest two concepts. Firstly, the sweat civilian medical practice. Surg. Clin. N. Am. 43, glands observed in this study could not have 5 I9-536. produced sweat. Therefore, hyperhidrosis as KATSAS A., AGNANTIS J., SMYRNIS S. & KAKAVOULIS T. (I977) Carcinoma on old frostbites. Am. J. a sequel to frost-bite may be more subjective Surg. 133, 377-378. than real. Secondly, the squamous metaLAPP N.L. & JUERGENS J.L. (I965) Frostbite. Mayo plasia seen in these sweat glands visually Clin. Proc. 40, 932-948. resembles a pre-neoplastic change. This MILLER D. & BJORNSON D.R. (i 962) An investiobservation suggests that frost-bite may be gation of cold injured soldiers in Alaska. Milit. one of the unproven influences of squamous Med. 127,247-252. cell carcinoma and that the site of origin RossIs C.G., YIACOUMETTIS A.M. & ELEMENOGLOU J. (I982) Squamous cell carcinoma of the heel could be sweat gland epithelium as well as at site of previous frostbite. I.R. Soc. developing the epidermis. Med. 75, 715-7I8.
7I6
Acknowledgements This experiment was done at USARIEM, Natick, MA, USA. The author thanks Drs
SCHONING P. & HAMLET M.P. (I989) Experimental frost-bite in Hanford Miniature Swine I Epithelial changes. Br. J. Exp. Path. 70, 41-49. SHAFER J.C. & THOMPSON A.C. (I955) Local cold injury. Arch. Dermatol. 72, 335-347.
