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In Vivo Effects of Laser Radiation on the Skin of the Syrian Hamster*†
Preliminary and Short Report IN VIVO EFFECTS OF LASER RADIATION ON THE SKIN OF THE SYRIAN HAMSTER*t SAMUEL FINE, S.M., M.D., EDMUND KLEIN, M.D., SIDNEY FARBER, M.D., RONALD E. SCOTT, M.A.SC., Sc.D., ALBERT ROY, M.A. AND RICHARD E. SEED, PH.D. Potential biological implications of laser radiation in the visible and infra-rcd region of the electromagnetic spectrum (1) have been discussed by Townes (2) and Solon, Aaronson and Gould (3). The term "laser" is a contractioa of light amplification by stimulated emission of radiation.
primary laser emission wave length was 10.600 A. Light was pumped into the rod by a straight xenon flash tube, using an input of 4750 or 6850 joules to
obtain an output of 30 or 50 joules, respectively. The duration of the flash tube output light pulse was 1.6 milliseconds. The duration of the laser output pulse was of the order of 1 millisecond. The
from stimulated emission from excited atoms or
intensity of the focussed 70 joule laser radiation was sufficient to produce a perforation approxi-
molecules. This process is the reverse of absorption
mately 1—2 mm in diameter when directed against
and occurs when there is an excess of atoms in an excited state. A light wave of suitable frequency travelling through the excited atoms will be ampli-
a steel plate )ieo inches in thickness.
Townes (2) describes laser radiation as resulting
EXPERIMENTAL DATA
fied. A light wave between parallel reflecting
surfaces can build up into a coherent oscillation. An output beam is obtained if one of the reflecting surfaces is partially transparent (2). The emitted radiation has the following charac-
Two groups of 6 normal adult Syrian hamsters each were studied. The animals were anesthetized with nembutal. The abdominal skin was shaved
6. Stable frequency; 7. Continuous, modulated or pulsed emission. Ocular damage induced experimentally in rabbits by means of a ruby laser with an output of 0.1 joule/0.5 m see. pulse, emitted in a coherent beam, monochromatic at a wave length of 6943 A and 1 cm. in diameter, was recently described by Zaret et al. (4, 5).
abdominal wall. The immediate effects resembled those of a 2nd degree thermal burn extending over a roughly circular area approximately 3 mm—S mm in diameter with a mild erythematous areola
and exposed to single pulses of laser radiation. teristics: 1. High degree of monochromatieity; Each animal in the first group was irradiated with 2. High degree of coherency; 3. Small angle of a single pulse of approximately 30 j oules focussed divergence; 4. Small spot size; 5. High intensity; onto areas on the external surface of the anterior
approximately 1—2mm in width. In parallel studies
The studies reported in this communication
the cheek pouch was exteriorized and irradiated. The immediate reaction was similar to a moderately severe burn. All animals in the first group recovered completely from the immediate effects of exposure to laser radiation. During the subsequent observation periods of 21 days they behaved normally in regard to food and water intake and general activity. They tolerated in a normal manner such procedures as repeated anesthesia and removal of biopsy specimens. The sites of irradiation underwent complete recovery whether the affected area was removed by biopsy-excision and sutured or whether the lesion was allowed to heal spontaneously. The gross appearances of the lesions in the skin on the 5th day post-irradiation suggested a more severe reaction in the lighter colored areas than in darker regions. This was also reflected in
were carried out to determine the effects of laser radiation on skin. Preliminary data are presented below. METHODs AND MATERIALS
The studies described here were carried out with lasers employing rods of neodymium-doped glass
capable of generating an output in excess of 70 joules. Each rod consisted of a core 0.250 inch in
diameter and 15 or 18 inches long, clad with a clear glass cladding 0.062 inch in thickness. The * From the Department of Electrical Engineering, Northeastern University, Boston, Mass., the
microscopic appearance, which presented Department of Dermatology, Roswell Park the fewer hair follicles in the lighter areas than in the Memorial Institute, Buffalo, N. Y., and the Children's Cancer Research Foundation, the darker areas, coagulation necrosis, leukoeytie inChildren's Medical Center, and the Department of Pathology, Harvard Medical School, Boston, Mass.
t The laser used in these studies was provided
through the cooperation of Dr. 0. W. Richards and Mr. G. A. Granitsas of the Research Department of the American Optical Company, Southbridge, Mass. Received for publication December 26, 1962. 123
filtration between necrotic areas and viable tissue, and epithelialization proceeding from the margin of the lesion. The animals in the second group were exposed to an output of 50 joules. The immediate and subsequent effects appeared to be similar to those observed in the first group. One of the animals in the
second group died within 5 hours of irradiation.
124
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
The lesions in this animal appeared similar to those
in the remaining 5 animals which survived. At autopsy, a moderately severe reaction consisting of hemorrhage and edema was observed, extending
through the anterior abdominal wall onto the pcritoneal surface without apparent discontinuity of peritoncum. Subjacent areas of the stomach and intestine showed distension. Subserosal hemorrhages and edema and subepithelial hemorrhages
at the corresponding luminal aspects of the
REFERENCES 1. Sciiwow, A. L. AND TowNEs, C. H.: Infrared
and optical masers. Phys. Rev., 112: 1940, 1958. 2. TowNEs, C. H.: Optical masers and their possible applications to biology. Biophys. J., 2: part 2, 325, 1962. 3. Soio, L. R., AARON5ON, R. AND GoULD, G.:
Physiological implication of laser beams.
Science, 134: 1506, 1961. 4. ZARET, M. M., BREININ, G. M., SCHMIDT, M., Rises, M. AND SIEGEL, I. M.: Ocular lesions
stomach and intestinal wall, respectively, were produced by an optical maser (Laser). Science, 134: 1525, 1961. present. M. M.: 23rd Meeting of the SubcomThe other 5 animals in the second group con- 5. ZARET, mittee on Atmospheric and Industrial hytinued to recover uneventfully during the regiene, National Academy of Science, Washmainder of the observation period.
ington, D. C., November 20, 1962.
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
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.
primary laser emission wave length was 10.600 A. Light was pumped into the rod by a straight xenon flash tube, using an input of 4750 or 6850 joules to
obtain an output of 30 or 50 joules, respectively. The duration of the flash tube output light pulse was 1.6 milliseconds. The duration of the laser output pulse was of the order of 1 millisecond. The
from stimulated emission from excited atoms or
intensity of the focussed 70 joule laser radiation was sufficient to produce a perforation approxi-
molecules. This process is the reverse of absorption
mately 1—2 mm in diameter when directed against
and occurs when there is an excess of atoms in an excited state. A light wave of suitable frequency travelling through the excited atoms will be ampli-
a steel plate )ieo inches in thickness.
Townes (2) describes laser radiation as resulting
EXPERIMENTAL DATA
fied. A light wave between parallel reflecting
surfaces can build up into a coherent oscillation. An output beam is obtained if one of the reflecting surfaces is partially transparent (2). The emitted radiation has the following charac-
Two groups of 6 normal adult Syrian hamsters each were studied. The animals were anesthetized with nembutal. The abdominal skin was shaved
6. Stable frequency; 7. Continuous, modulated or pulsed emission. Ocular damage induced experimentally in rabbits by means of a ruby laser with an output of 0.1 joule/0.5 m see. pulse, emitted in a coherent beam, monochromatic at a wave length of 6943 A and 1 cm. in diameter, was recently described by Zaret et al. (4, 5).
abdominal wall. The immediate effects resembled those of a 2nd degree thermal burn extending over a roughly circular area approximately 3 mm—S mm in diameter with a mild erythematous areola
and exposed to single pulses of laser radiation. teristics: 1. High degree of monochromatieity; Each animal in the first group was irradiated with 2. High degree of coherency; 3. Small angle of a single pulse of approximately 30 j oules focussed divergence; 4. Small spot size; 5. High intensity; onto areas on the external surface of the anterior
approximately 1—2mm in width. In parallel studies
The studies reported in this communication
the cheek pouch was exteriorized and irradiated. The immediate reaction was similar to a moderately severe burn. All animals in the first group recovered completely from the immediate effects of exposure to laser radiation. During the subsequent observation periods of 21 days they behaved normally in regard to food and water intake and general activity. They tolerated in a normal manner such procedures as repeated anesthesia and removal of biopsy specimens. The sites of irradiation underwent complete recovery whether the affected area was removed by biopsy-excision and sutured or whether the lesion was allowed to heal spontaneously. The gross appearances of the lesions in the skin on the 5th day post-irradiation suggested a more severe reaction in the lighter colored areas than in darker regions. This was also reflected in
were carried out to determine the effects of laser radiation on skin. Preliminary data are presented below. METHODs AND MATERIALS
The studies described here were carried out with lasers employing rods of neodymium-doped glass
capable of generating an output in excess of 70 joules. Each rod consisted of a core 0.250 inch in
diameter and 15 or 18 inches long, clad with a clear glass cladding 0.062 inch in thickness. The * From the Department of Electrical Engineering, Northeastern University, Boston, Mass., the
microscopic appearance, which presented Department of Dermatology, Roswell Park the fewer hair follicles in the lighter areas than in the Memorial Institute, Buffalo, N. Y., and the Children's Cancer Research Foundation, the darker areas, coagulation necrosis, leukoeytie inChildren's Medical Center, and the Department of Pathology, Harvard Medical School, Boston, Mass.
t The laser used in these studies was provided
through the cooperation of Dr. 0. W. Richards and Mr. G. A. Granitsas of the Research Department of the American Optical Company, Southbridge, Mass. Received for publication December 26, 1962. 123
filtration between necrotic areas and viable tissue, and epithelialization proceeding from the margin of the lesion. The animals in the second group were exposed to an output of 50 joules. The immediate and subsequent effects appeared to be similar to those observed in the first group. One of the animals in the
second group died within 5 hours of irradiation.
124
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
The lesions in this animal appeared similar to those
in the remaining 5 animals which survived. At autopsy, a moderately severe reaction consisting of hemorrhage and edema was observed, extending
through the anterior abdominal wall onto the pcritoneal surface without apparent discontinuity of peritoncum. Subjacent areas of the stomach and intestine showed distension. Subserosal hemorrhages and edema and subepithelial hemorrhages
at the corresponding luminal aspects of the
REFERENCES 1. Sciiwow, A. L. AND TowNEs, C. H.: Infrared
and optical masers. Phys. Rev., 112: 1940, 1958. 2. TowNEs, C. H.: Optical masers and their possible applications to biology. Biophys. J., 2: part 2, 325, 1962. 3. Soio, L. R., AARON5ON, R. AND GoULD, G.:
Physiological implication of laser beams.
Science, 134: 1506, 1961. 4. ZARET, M. M., BREININ, G. M., SCHMIDT, M., Rises, M. AND SIEGEL, I. M.: Ocular lesions
stomach and intestinal wall, respectively, were produced by an optical maser (Laser). Science, 134: 1525, 1961. present. M. M.: 23rd Meeting of the SubcomThe other 5 animals in the second group con- 5. ZARET, mittee on Atmospheric and Industrial hytinued to recover uneventfully during the regiene, National Academy of Science, Washmainder of the observation period.
ington, D. C., November 20, 1962.
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
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.