Particulate environmental matter as seen in lung sections

PARTICULATE ENVIRONMENTAL MATTER AS SEEN IN L U N G S E C T I O N S Roger A. Schinella, M.D.

Abstract A study o f particulate e n v i r o n m e n t a l m a t t e r as seen in lung sections is presented. T h e s e particles are blackened and are c o m p a r a b l e to particles seen in industrial-urban environments. From the study o f histologic sections an attempt is m a d e to d e t e r n t i n e the tissue reaction to them and their uhimate fate.

In examinations o f autopsy and surgical sections o f h m g a variety o f particles were seen in the tissue. T h e p u r p o s e o f this presentation is to describe these structures, postulate as to their origin, and describe the tissue response to them. O u r study c o n c e r n e d the relatively large (up to 80 to 90/.t) atnmspheric particulate pollutants f o u n d in tile hulnan lung. We thouglat that a stud)" o f such structures could p r o d u c e i n f o r m a t i o n about the physiologic response o f lung tissue to large particles mad p e r h a p s be o f help in specific environmental or industrial situations in whicli sttch particles occur in large nuntbers ~.

MATERIALS A N D METHODS 1. Dtn'ing routine exantination o f histologic sections o f lung front autopsy and surgical cases, note was m a d e o f the appearance o f these particles. Front these sections 23 autopsy and two surgical lung specimens were selected for study. In three o f the autopsy and one o f the surgical cases multiple serial se~:tions were obtained. Spe-

ciai stains, including periodic a c i d - S c h i f f , l'erls, Masson trichronm, and VerhoeWs elastic stains, were used as well as hematoxylin and eosin stain. Staining was carried out on consecutive n u m b e r s o f the serial sections. All sections were viewed with a polarizing naicroscope. Pediatric autopsy lung sections f r o m 10 cases were also reviewed. T h e patients r a n g e d in age front fotu" to 14 years, ht all cases clinical histories were obtained. '2. Slide adhesive-air sample specimens were obtained by covering most o f the surface o f a s t a n d a r d glass slide with a liquid commercial slide adhesive preparation.* T h e slide containing this lnaterial was then put on the sill o f one o f the windows o f the surgical l)athology laboratory o f Bellevue Hospital facing a busy city avenue. T h e slides were collected at intervals ranging fl-om o n e - h a l f to three hours. T h e dried or partially dried adhesive preparation along with all a d h e r e n t air dust was scraped off and sublnitted fin" histologic *Tissue-Tac, Dade Division, American l lospital Suppl)" Corp., Miamj, Florida 33152.

*Assistant Professor of l'athology, New York University School of Medicine, New York, New York.

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processing in the m a n n e r o f a cell block. T h r e e to nine subserial sections were obtained for each sample, nmking possible a r o u g h histologic examination o f particulate m a t t e r in the air. Six such samples were e x a m i n e d d u r i n g the winter and spring o f 1974. T h e y were obtained during periods o f good and p o o r air quality. 3. L u n g sections f r o m six cases o f s u d d e n or rapid death (by courtesy o f the New York Medical Examiner's office) were e x a m i n e d for these particles. T h e s e sections were f r o m individuals who died within minutes as a r e s u h o f homicide, suicide, o r accidents. 4. Twenty-five Papanicolaou and 25 "cell block" examinations o f s p u t u m were a l s o e x a m i n e d for these particles.

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March 1976

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RESULTS

T h e first category o f particles seen consisted o f lace)" o r bubbly, fenestrated fragments (Fig. 1). T h e s e were often f o u n d

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in m a c r o p h a g e s and almost always f o u n d in ah'eoli. In o n e instance they were f o u n d inside a m a c r o p h a g e that was located in a bronclfiole. Rarely these structures were f o u n d in the p u h n o n a r y interstitial tissue and in these cases were witlfin macroplmges or giant cells. T h e y were always o f a d e e p black color a n d r a n g e d in size f r o m 2 to 40 it. T h e second type o f particle was spheroidal. T h e spheroidal form was d e m o n strated b)" serial sections, which showed consecutively a plate merging into a ring tlmt became larger and then smaller, and finally a plate again. T h e walls o f these structures were often irregular or frayed (Figs. 2, 6A). Defects were often present in the walls. Most often a r o u n d or witlfin these structures macroplmges o r giant cells could be fotmd. W h e n the spheroidal structure was witlfin a m a c r o p h a g e o r giant cell, tlmt portion o f the cell cytoplasm was often _pale staining. W h e n the particles were outside cells, there was often a granttiar or fibrillar material inside the particles

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where alveolar nmcrophages were numerous. The majority of tlte patients, except in cases of rapid deatlt reported from the Medical Examiner's office, had been on respirators or had had prolonged intubation, had received oxygen, or had been in coma for prolonged periods. Examinations of slide adhesive-air sample preparations showed particles of similar size and similar lfistologic appearance (Figs. 5, 6B). T h e lacey-bubbly particles and jagged particles were found more readily than the spheroidal particles. In fact, the latter were found in only one of tlte 27 sections made fi-om the slide adhesive-air samples. All the forms of particles were found in sputunt specimens. Tltey were found more readily in cell block preparations of sputum than in sputunt smears. The particles tended to be smaller than those seen in lung sections. They were found in macrophages but were more often free in the mucus material. Structures sinfilar to all the types we

Figure 3. A spheroidal structure within a corpus amylaceum. (ttematoxylin and eosin stain, x400.)

->r ! that appeared to be inspissated proteinaceous material. The structures were also found rarely in alveolar septa and in corpora amylacea (Fig. 3). They were not found in bronchi or bronchioles. Their sizes ranged from 4 to 90/.t. The third particle consisted of jagged, platelike structures, which were uniformly black and irregularly sltaped. Tltey ranged in size from 2 to 30/z (Fig. 4). None of these particles were found in hilar lymph nodes. Periodic acid-Schiff, l, erls, \ erhoeff elastic, and Masson tricltrome stained sections could not be adequately interpreted because of the intense black color of these particles. Tire particles showed slight birefringence when viewed witlt polarized light and were slightly refractile when viewed witlt reduced light. The particles were demonstrable even in areas where extensive lung destruction had occurred, as in infarcts, severe necrotizing pneumonias, and witifin areas of caseation necrosis. The particl~:s were found most often in hmg sections showing patltologic aherations, as from pneumonia, and in areas

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225

HUMAN PATHOLOGY--VOLUME 7, NUMBER 2 March 1976 have described were seen in the l u n g sections o f t h r e e o f tim ten children e x a m i n e d bttt were few in m t m b e r . ( T h e ages o f tim children were 13, 10, a n d five years.) Similar strttctures were also seen in two o f the six cases o f r a p i d violent death. In these two cases they were a b u n d a n t ; both were cases o f asphyxia, o n e by b a n g i n g a n d one by ligature strangulation.

DISCUSSION T h e r e was a m o r p l m l o g i c similarity between the structures seen in l u n g sections, in slide adhesive-air s a m p l e p r e p a r a tions, a n d in cell blocks o f s p u t u m specin l e n s . S o m e o f these s t r t t c t u r e s w e r e also similar to particles described in certain in-

Figure 5. Section obtained from a slide adhesive-air sample. Note the similarity of this structure to that in Figure 1. (Hematoxylin and eosin stain. •

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PARTICULATE MAqq'ER IN LUNG SECTIONS--SclIISELLA dustrial and u r b a n environments; for example, the lacey-bubbly black particles seen in o u r sections were similar in a p p e a r a n c e and size to particles o c c u r r i n g in oil soot as described in The l'article Atlas by McCrone et al. T h e platelike structures f o u n d in the h m g sections were similar to those described as coke and coal combustion products. z T h e y were also within tim range o f particle size described for [ly ash a and app e a r e d to be similar to the hemosiderin encrusted particles described by Gross et al. 4 as ferruginous bodies with a black central

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T h e equivalent o f the spheroidal structures seen in the h m g sections was not f o u n d in The Particle Atlas, yet they were commolfly seen in h m g sections. A similar structure was f o u n d , but only once, in a slide adhesive-air sample specimen. Similar spheroidal particles were discussed in a case presented at a seminar on h m g diseases as a probable atypical acid fast infection o f the lung. ~ In this lung great n u m b e r s o f the spheroidal particles were also described. T h e patient was a steel worker. T i m structures were described as a p p e a r i n g to excite a macroplmge and fibrous response and their o u t e r rims as a p p e a r i n g birefi'ingent as t h o u g h they were coated with calcium o r iron. T h e case was designated as "crystalline p n e u n t o p a thy" but tim structures were not f n r t h e r defined. Gross et al. 4 also described and illustrated a similar "black ring" inside a ferruginous body. T h e s e and similar black particles encrusted by hemosiderin were t h o u g h t to be products o f plant combustion. Dr. . ! l'aul Gross, who kindly reviewed the secttbns containing some o f these particles, felt that tim larger, fenestrated, laceybubbly particles were incompletely b u r n e d vegetable or plant products, whereas the spherical or ctLrved structures were derivatives o f h y d r o c a r b o n combustion. U n f o r t u n a t e l y PAS as well as o t h e r special stains that might have helped to d e t e r m i n e whetlter some o f these were ind e e d vegetable particles could not be interpreted because o f the d e e p black coloration o f these structures. Nevertheless septate forms o f the spheroidal structures and their slight r e f r a c n h t y and bn-efringence strongly suggest a vegetable or plant origin (Fig.

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As m e n t i o n e d , the spheroidal particles were rare in slide adhesive-air sample specimens ancl are not illustrated in The l'article Atlas. Yet they were readily f o u n d in many lung sections selected at r a n d o m a n d ill s o n l e c a s e s w e r e qnite n t l n l e r o u s . Several explanations can be offered to explain tiffs. I. T h e y are t r a n s t o r m e d structures; that is, they may r e p r e s e n t lacey-bubbly or septate structures that the body has b r o k e n down to spheroidal structures. 2. T h e y are p r e s e n t in the environment but are less c o m m o n and tim lung tends for some reason to concentrate them. 3. Although t h e r e is evidence that these structures are e n v i r o m n e n t a l in origin, there is the possibility tlxat the spheroidal structure is mamnfactured by the bocly. In a recent publication an electron micrograpll is shown o f an intracytoplasmic structure f o r m e d b~" carbon material e n c r u s t e d on the m e m b r a n e o f a lysosome.6 T h e size o f the s t r u c t n r e illustrated is 1 to 3/*. H o w e v e r it lxas been d e m o n s t r a t e d that lysosomal structures can attain large sizes u p to 50/.t a r o u n d asbestos particles, t

227

HUMAN I'ATHOLOGY--VOI.UME 7, NUMBER 2 Mmrh 1976 scribed b)' McCormack, ~ often where the normal lung p a r e n c h y m a had been obliterated by fibrosis. T h e y were also f o u n d intact in areas o f necrosis, such as infarcts, in areas o f caseation, and within areas o f necrotizing p n e u m o n i a . "Fhe fact that these particles were often f o u n d in macrophages indicates that they were treated as "foreign." I'erhaps the fact that these probably lbreign e x o g e n o u s structures are f o u n d in c o r p o r a amylacea and in ferrttginotts bodies and are e n t r a p p e d in areas o f fibrosis may indicate that the body is forced to effect encrustation b)' protein or hemosiderin or e n t r a p m e n t by fibrotts tissue as a means o f dealing with these particles. T h e e n v i r o n m e n t a l particle size for maximal ah'eolar deposition is calculated to be a p p r o x i m a t e l T 2p.3 Only a small proportion o f e x o g e n o u s particles in the r a n g e o f 2 to 90/z n o t e d in this stud)" are t h o u g h t to reach the terminal bronchioles and pulmonar)" l~arenchyma u n d e r normal conditions. T h e majority o f these larger particles are fihered out in the tmsoplmrynx or

Examination o f sptttum specintens smears and cell blocks was directed toward d e t e r m i n i n g w h e t h e r these particles are elinfinated via the sputuna. T h e particles were seen in sputuna specintens. T h e fact that the majority were not in macroplmges suggests that most were probably being r e m o v e d by "impaction" in bronchi. In h m g sections one o f the lacey-bubbly particles was seen in a macrophage, which in turn was in the lumen o f a bronchiole, suggesting that it was exiting via the bronchiole f r o m the ah'eolus where phagocytosis had taken place. Evidence was f o u n d , however, that these structures achieved some degt+ee o f p e r m a n e n c e in the lung. The)" were f o u n d in c o r p o r a amylacea, indicating protein encrustation on the particles as a nidus. As m e n t i o n e d previottsly, Gross et al. a described spheroidal and platelike carbonaceotts particles encrusted by hemosiderin and f o r m i n g ferruginotts bodies. T h e s e particles were also t b u n d within areas o f fibrosis in this study and in the case de-

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PARTICULATE MATI'ER IN LUNG SECTIONS-ScnlxrlaA in the upper respiratory passages, a Tire particles noted in tltis study nmy represent a minor proportion of particles titat do filter down to the pulmonary parencltyma. In a majority of cases tire patients whose tissues were examined in this study were comatose; some required tracheostomies or endotraclteal intubation or received oxygen-air mixtures by respirator, mask, or nasal catlteter. These procedures of course would bypass the nasal and t, pper respiratory tract filters responsible for fihering many of the large particles? However, it ntust be noted that even in cases of death by asphyxia these particles were found (albeit in small ntunbers), indicating that even witltout ntechanical bypass of nornm! physiologic air filters these particles do reach rite lung parencltyma. Tire particles were found somewhat more frequently in areas of tire lung Where a greater number of intralveolar macropltages were found, possibly because ntacropltages are ntobilized in response to the particles or material carried on thent (for example, toxic substances absorbed on the particles). Anotlter possibility is that since many of tltese patients were receiving exogenous oxygen, tire increased nuntber of macroplmges is a response to the oxygen as an irritant. Ahernatively, these particles may tend to be deposited in areas of lung that have been danaaged, thereby decreasing the cap,tcity of tlte tissue to remove the particles in both the upper respiratory passages subtending that area of lung and in the lung p,trenchynm itself. The histolugic appearance of tltese particles may be simulated by several otlter strttctures. For example, air bubbles with encrusted formalin pigntent arofind thent may rdsentble those particles (Fig. 8A). Second, the particles are actually present in the air and may at any tinte during the processing of tissue contaminate it. WitCh the latter occttrs, they may be found in the sante plane of section as the tissue and ma)' be difficult to distinguish from particles present in the tissue in vivo. Finally in the lung it was noted during this study that at times macrophages formed ringlike aggregates around proteinaceotts material (Fig. 8B). Tltese may simulate sections of tire splteroidal particles. Indeed early in the investigation sucit particles were titougllt to be possible precursors of the splterical

particles. However, because careful examination never revealed a black color in any of these aggregates, tltis possibility was rejected. No relation was noted between the concentration of these particles and tire occupational or smoking Itistory or the presence of cin'onic lung disease. These particles not only were found in patients from the Bellevue Hospital population but were also found in lung sections from patients in suburban New Jersey and the Sacramento, California, area.

ACKNO~VLEDGMENTS

Thanks are due to Miss Rosalyn Klein for typing the ntanuscript and to Mr. Robert Waldeck for iris photograplfic assistance, as well as to I)octors Paul Gross, Sidney Laskin, Marvin Kuschner, Philip Prose, and Michael Baden for assistance and suggestions.

REFERENCES 1. McCrone, W. C., Dnffty, P,. G., aqd I)elly,.]. G.: Tile Particle Atlas. Ann Arbor, Michigan, Ann Arbor Science Publishers Inc., 1973, Vol. 2, Figs. 391 to 393. 2. McCrone, W. C., Draft)', R. G., and I)eIly, J. G.: T h e Particle Atlas. Ann Arbor, Michigan, Ann Arbor Science Publishers Inc., 1973, Vol. 2, Fig. 382. 3. Stuart, B. O.: Deposition of inhaled serosals. Arch. Int. Med., 131:60, 1973. 9t. Gross, 1'., Tulna, J., and deTreville, R. T. i'.: Unusual ferruginous bodies: their formation from nonfibrous particulates and from carbonaceous fibrous particles. Arch. Environ. ttealth, 22: 534, 1971. 5. McCormack, k J.: In discussion of Case 5, crlistalline pneumopathy, 21st Ammal Seminar, Indiana Association o f l'athologists, 1971. 6. liarrison, G., and Lapp, N. L.: Electron microSCOlfiC findings in the hmgs of miners. Ann. N.Y. Acad. Sci., 200:73, 1972. 7. Suzt,ki, Y., and Churg, J.: Structure and development of the asbestos body. Amer. J. Path., 55: 79, 1969. 7. Goldberg, I. S., and l.ouren~o, R. V.: Deposition of aerosols in pulmonary disease. Arch. Int. Med., 131:88, 1973. I)el)artnlent of Pathology New York University School of Medicine 550 First Avemle New York, New York 10016

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