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Effects of Melanin Pigment on Target Specificity Using a Pulsed

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Effects of Melanin Pigment on Target Specificity Using a Pulsed Ultrastructure: Effects of Melanin Pigtt1ent on Target Specificity Using a Pulsed Dye Laser (577 ntt1) Arthur K. F. Tong, M .B., B.S. , Oon T. Tan, M.D., James Boll , M.Eng., John A. Parrish, M .D., and George F. Murphy, M.D. Dcp:utmmt of Dermatology, Boston U ni versity School of Medicin c (OTT), Boston, Dcpartmcnt of Pathology, Brigha l11 Jnd Worncn's Hospital (G FM), Boston, and Dcpartment of I ermatology, Massachusctts Gencra l Hospital (AKFT, JB, JAP), Boston, Massachusetts, U.S.A. It h as been shown recently that brief pulses of 577 nm doses. We conclude that epidermal mela ni n and vascul ar radiation fro m the tunable dye laser are absorbed sel ectively hem oglobin are competing sites for 577 nm laser absorp­ by oxyhemoglobin. T his absorption is associated with highl y ti on and damage, and that the target specifi city of the 577 specifi c damage to superficial vascul ar pl exus blood vessels nm tunable dye laser is therefore inAuenced by variations in those with li ghtly pigm ented (type I- II) skin . in epidermal pi gmentation. T hi s finding is rele va nt to the To determine w hether pigmentary differences in the clinical applica ti on of the tunable dye laser in the ablative overlyin g epidermis inAuence this target specifi city, w e trea tment of vascul ar lesions. exposed both type I (fair) and type V (dark) normal human We also fo und on ultrastru cture that the presence of elec­ skin to varyin g radiant exposure doses over 1. 5-fLS pulse tron-lucent circular structures of approximatel y 800 A in durations from the tunable dye laser at a wavelength of577 di am eter were observed onl y at and above cl in ica l threshold nn~. U sing ultras tructural techniques, we found in type I doses in those with type I skin and at the highes t dose of skin that even clinica l subthres ho ld laser exposures ca used 2.75 JI cm 2 in type V skin. I t has been proposed that these reproducible alterations of erythrocytes and adj acent der­ structures might be hea t-fixed m olds of water vapor. Both mal vascular endothelium w ithout comparable damage to this and ul tras tructural changes o f epidermal basa l cells the overl yin g epidermis. In contras t, degenerated epider­ demonstrate m echanisms responsible for alteration of ti ss ue mal basal cel ls represented the predomin ant form of cellular after exposure to 577 nm, w hi ch arc di scussed. J fillies/ d aITlage aft er laser exposure of type V skin at comparabl e D erll/a/o l 88:747-752, 1987 lthough lasers have been in medica l use fo r about 20 and morc w id espread ti ss ue injury results. Therefore, in order to years as ca utery and cutting devices, the multitude produce highl y selective thermal damage to pigmented tissue tar­ of facto rs that determine the o utcome of laser-tissue gets such as bl ood vessels or melanin-containing cell s, very shorr interactions are still being defin ed. In general, a com­ and intense pulses at selectively absorbed wavelengths must be bin ati on of intera cti ons occurring at the ti me of laser ad ministered. Aexposure determine the sites, nature, and extent of immediate It is now possible to select laser parameters w hich res ul t in tissu e injury, w hi ch then initiates repair processes. Absorption of exqui sitely selective thermal damage to vessels or to pigmcnt cell s lase[' cnergy in sufficient quantity at given ti ss ue sites, such as [1 -3 J. Y cll ow (577 nm) pulsed dye lasers have recently been used blood vessels and pi gmented cell s, succeeds in heatin g these and to ca use hi g hl y specific injury to the cutaneous mi crovascul ature. o the[' nearby structures to denaturing or vaporizing temperatures. C linica l applica tions of such selective vascul ar injury include treat­ The extent to which thermal damage is precisel y locali zed to sites ment of port-wine les ions [4,5J, telangiectases, and other vascular of selecti ve absorption is largely determined by w hether the laser abnormalities and tumors. The 577 nm wavelength is strongly exposure is sufficiently short to prevent thermal diffusion fr om absorbed by oxyhemoglo bin, but there is also absorption by epi­ these sites . If diffusion occurs, surrounding ti ss ues are also heated, dermal melanin at this wavelength [6]. The overl yin g epidermal pi gment layer, therefore, represents a potentia l barrier thro ugh w hi ch 577 nm rad iati on must pass to reach the underl ying m i­ Manusc ript received September 2, 1986; accepted for publi ca tion De­ crovascul ature. This is evidenced by an in crease in the radiant cember 3 1, 1986. cxposure dose necessary to ca use vascul ar damage w ith in creasing T h is stud y was supported in part by the Arthur O. and Gullan M . mela nin pigment [6]. Thus, it is possible that individuals with Wellman Fo undation and by Mary Kay Dermatology Foundation Fel­ darker skin are less sensitive to the selective vascul ar damage lowship awarded to O. T. Tan, M .D. ca used by 577 nm pul ses . Reprint requests to: Oon T. T '1I1 , M . D ., Department of Dermatology, In ord er to characteri ze the cffects of epidermal melanin pig­ Boston Unive rsity School of Medi cin e, 80 East Concord Street, Boston, Massachuse tts 0211 8-2394. m entati on on sel ective vas cular inj ury, we exposed skin ofhul11 3n A b brevia tions: subjects of differing constitutive pigmentation to a pulsed tunable RUC: red bl ood cell (s) dye laser at 577 nm and studied the morphologic and ultrastruc­ T: threshold dose tural effects produced over a range of radi ant exposures. 0022-202X/87/S03.S0 Copyri ght © 1987 by The Society for In ves ti gative Dermatology, In c. 747 748 TONG ET AL THE JOURNAL OF INV ESTIGATIVE DERMATOLOGY MATERIALS AND METHODS Buttock skin of 4 healthy human male adult volunteers (3 with skin type I and 1 with skin type V) was exposed to varying radiant exposure doses of laser radiation from a Candela Model #LFDL 1 tunable dye laser operating at 577 nm usin g a rhodamine 575 dye at a pulse duration of 1.5 f.L s. The laser was fo cused usin g a pl anoconvex lens, into a quartz optical fib er. Laser pulse energies were meas ured usin g a Scientech laser energy meter calibrated to ± 10% accuracy. Radiant exposure dose was ca lculated based on the diameter of the circular fi eld of irradi ati on. Separate 3 mm­ diameter sites were exposed to sin gle laser pulses at flu ences rang­ 2 2 2 in g from 0.5 Jlcm to 2.75 Jlcm , in 0.25 J/c m in crements. C linica l (morphologic) thres hold was defin ed as the presence of nonblanchable purpura filling the entire exposed site of 3 mm diameter, appearin g within 10 min of laser exposure. The energy required to produce this effect was defin ed as the threshold dose (T). Four 3-mm punch biopsies were taken from each individual at predetermined radiant exposure doses after infiltration of the skin with 1 % Xylocaine without epinephrine. All biopsies were ob­ tained within 10 min of laser irradiation and split for li ght mi­ croscopy (routine H & E staining) and electron microscopy. The tissue was processed for transmission electron microscopy as pre­ viously described by Murphy et al [7] . Ultrathin sections were Figure 1. Superfi cial capi ll ary venule of type I skin after exposure to 2 cut w ith a diamond knife on a Porter Blum MT-2 ultra­ subthreshold dose (0.5 J/cm ) of laser irradiation at 577 nm. N ote the microtome, counterstained w ith uranyl acetate and lea d citrate, marked swelling of endotheli al cells (a sterisks) producing narrowing of and examined with a JEOL JSM 100S electron microscope. the vascular lumenal diameter (L) . Focal gaps (a rrow) between adjacent endothelial cells (EC) and retraction of the endothelial cell s (arrowhead) RESULTS away from pericytes (P) can also be seen. There was a direct relationship between laser energy required to produce the morphologic effect of clinica l purpura and skin pig­ mentation (Table .I); a mean dose of 1 J/c m 2 (range, 0.75-1.25 melan ocytes were also profoundl y altered and rough endoplas mic 2 J/c m ) was required to produce purpura in those with type I skin, reti culum within keratinocytes appeared dilated and mitochon­ whereas it was not possible to produce purpura in the skin type drial swelling was apparent (Table II ). Similar ultrastructural changes V subject, even at the maximum ava il able laser exposure of 2. 75 were also seen in Langerhans ce ll s. In spite of these changes, 2 J/c m . Li ght microscopic and ultrastructural examinati on ofbiop­ melanosomes, the basement membrane zone, and nuclei still ap- sies, taken from control (non-laser exposed) sites and processed under identi ca l conditions appea red normal, with intact vessels and a normal-appea ring epidermis [8]. Morphologic analyses of laser-exposed sites are given below. Table II. Compari son of Epidermal Alteration Between Skin Types I and V at Similar Laser Energies Type I Skin Although only minimal erythema was clinica ll y Laser Energy detected aft er exposure of type I skin to a subthres hold dose of 2 2 0 /cm ) Skin Type I Skin Type V 0.5 J/c m , ultrastructural examination of the epidermis at this energy showed the presence of small cytoplas mic vacuoles in basa l 1.
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