Document zzgy2wQJjk1LG3L3Z9D402Ep0

R&S 135114 9rulkutf/ii(il BIO -TEST JlaJx&jd&j&t,, 3-nc. 1810 FRONTAGE ROAD NORTHBROOK, ILLINOIS 60062 REPORT TO MANUFACTURING CHEMISTS ASSOCIATION GROSS AND HISTOPATHOLOGIC FINDINGS ONE-YEAR INTERIM SACRIFICE CHRONIC VAPOR INHALATION STUDY WITH VINYL CHLORIDE (ETHYLENE DERIVED) IN ALBINO RATS MAY 9, 1975 IBT NO. 663-03222 9fV$JuM/lu2L BIO -TEST J)(7jyy*C`t&lie6-. 9-nc. 1810 FRONTAGE ROAD NORTHBROOK. ILLINOIS 60062 May 9, 1975 R&S 135115 Mr. Milton Freifield Project Manager, Vinyl Chloride Research Manufacturing Chemists Association 1825 Connecticut Avenue, N.W. Washington, D. C. 20009 Dear Mr. Freifield: Re: IBT No. 663-03222 - Chronic Vapor Inhalation Study with Vinyl Chloride (Ethylene Derived) in Albino Rats - Gross and Histopathologic Findings - One-Year Interim Sacrifice We are submitting herewith our laboratory report prepared in connection with the above study. Very truly yours. JCC: mcb Xp J. C, Calandra President Dkduttbual B I O - T E S T J.aJta'iaityued., Hhc. REPORT TO MANUFACTURING CHEMISTS ASSOCIATION GROSS AND HISTOPATHOLOGIC FINDINGS ONE-YEAR INTERIM SACRIFICE CHRONIC VAPOR INHALATION STUDY WITH VINYL CHLORIDE (ETHYLENE DERIVED) IN ALBINO RATS MAY 9, 1975 IBT NO. 663-03222 I. Introduction At the request of the Manufacturing Chemists Association, 40 rats (5 males and 5 females) from the control and test groups were sacrificed on September 5, 1974, to conduct cytogenetic studies on bone marrow specimens obtained at time of sacrifice. The purpose of this report is to present the gross and histopathological findings in other tissues from these animals. R&S 135116 !)ndudi-ual B 1 O - T E S T J.alt&iahyued., 9+tc. -4>> Vv' JvSry J*'*' J \ II. Summary L~ ! Treatment-related gross and/or histopathological findings were confined ", to the liver of test animals at exposure levels of 50, 200 and 2,500 ppm. C ^ ^O /' Gross pathologic findings ascribed to vinyl chloride were confined to ^ the liver of 2 animals of the 200 ppm level (group T-II) and the liver of 5 ) -animals of the 2,500 ppm level (group T-IH) . Findings consisted of solitary, .J ? ^* ^ ; `(dark red, nodular,foci in the subcapsular region of single liver lobes. These \vr* J J y r nodules varied in size from approximately 4 mm to 4 cm in diameter and, on cross-section, they contained blood-filled cystic spaces. Upon histopathological examination, the above lesions were identified as sites of hepatic angiosarcoma. In addition, there was focal hypertrophy and /' hyperplasia of hepatocytes in the liver sections from test animals at each exposure level. The incidence and relative severity of this finding could be correlated with the level of exposure to the test material. No extra-hepatic angiosarcomas were observed. Materials and Methods All animals, selected at random for sacrifice, were rendered unconscious JJ Co ..in a closed chamber by exposure to carbon dioxide gas and exsanguinated, and CO co cn a complete necropsy examination was performed. The organs were weighed. Representative specimens of the following organs and tissues were placed in 10% neutral buffered formalin for fixation; DndmiuaL B I O - T E S T ^alMyiaiyueS, 2*tc. 3 R&S 135118 Adrenal Glands All Gross Lesions Bone (femur and sternum) Bone Marrow (sternal) Brain and Spinal Cord Esophagus Eye Gonads (testes and ovaries) Kidneys Large Intestine (caecum and colon) Liver Lungs Lymph Nodes (tracheobronchial, cervical and mesenteric) Optic Nerve Pancreas Parathyroid Gland Pituitary Gland Prostate Gland Salivary Gland Seminal Vesicles Skin Small Intestine (duodenum, jejunum and ileum) Spleen Stomach Thymus Thyroid Gland Trachea Urinary Bladder Uterus The above tissues from 'all animals were processed by conventional methods, embedded in Paraplast*, sectioned at 4-6 microns, stained with Hematoxylin and Eosin and examined by light microscopy. The gross pathologic findings are presented in Tables I through II. * The histopathological findings are presented in Tables III and IV. The pathologist's report is presented on pages 4 through 6. Respectfully submitted. INDUSTRIAL BIO-TEST LABORATORIES, INC. Report prepared by: Donovan E. Gordon, D.V.M., Ph.D. Section Head, Pathology Report approved by: Manager, Toxicology mcb * Sherwood Medical Industries, St. Louis, Missouri 63103. jndudbUal B I O - T E S T 9*tc, 4 IBT No. 663-03222 Manufacturing Chemists Association February 27, 1975 R&S 135119 A gross and histopathological examination was conducted on a series of tissues and organs from rats of the 12-month interim sacrifice of IBT No. 663-03222. Treatment-related gross pathologic changes were confined to the liver of 2 T-II females (Nos. 550 and 599), 1 T-III male (No. 694) and 4 T-III females (Nos. 717, 732, 735 and 742). The lesions consisted of solitary, dark red nodules within the subcapsular region of 1 liver lobe. The nodules measured approximately 4 mm to 4 cm in diameter and contained blood-filled cystic spaces. There were fibrous adhesions between the capsular surface of some of these nodules and adjacent visceral structures. In 1 animal (No. 735), there were 2 different liver lobes with solitary nodular foci similar to the above. Upon histopathologic examination of the hepatic nodules, they were found to be angiosarcomas. A small amount of hemorrhagic fluid was present in the abdominal cavity of 1 animal (No. 59 ) as a result of hemorrhage from the surface of the liver tumor. Treatment-related histopathologic findings were confined to the liver of 5 T-I animals and all of the T-EI and T-III animals. There was an exposurerelated incidence and severity of hypertrophy and hyperplasia of hepatocytes throughout the.Ji.ver-sections . The hypertrophied hepatocytes had an increase in cytoplasmic mass which had a light, eosinophilic staining appearance with a few basophilic condensations. In some sections, these cells formed small ^ndu&buaL B I O - T E S T Xalwaiiyiiei, Dnc. 5 nodules with very little compression of the adjacent parenchymal cells of the liver. Some of the hyperplastic hepatocytes had prominent nuclei and nucleoli. They also formed small, discrete nodules in some areas. In addition, these cells often had a slightly increased cytoplasmic basophilia. In regard to the angiosarcomas, the early and more advanced stages of the tumor could be identified. In the early lesion, there was focal sinusoidal dilatation accompanied by hyperplastic and/or hypertrophied endotheliallike lining cells. The areas usually communicated with frank areas of angiosarcoma. The latter were characterized by the presence of blood-filled cystic sinusoidal spaces (peliodc foci) in which there were papillary pro jections of cords of liver cells that were covered by large, hyperchromatic to pleomorphic lining cells on the sinusoidal surface. The hepatocytes in the area adjacent to the tumor were also invaded by these tumor cells. In some areas, there was complete destruction of the enveloped liver cords and replacement by solid sheets of sarcomatous tumor cells which contained numerous mitotic figures and had anaplastic features. In most instances, the tumors were situated just beneath the capsular surface of the affected liver lobule and there was focal invasion of tumor cells into the capsule with hemorrhage into the peritoneal cavity at these sites. There were also fibrous adhesions to the adjacent visceral organs at the capsular surface of the tumors which correlated with the gross observations. In 2 animals (Nos. 663 and 668), the tumor foci were too small to be detected microscopically and they were in the early stages of development. In this study, there were no metastatic R&S 135120 BIO-TEST 9*tc. 6 tumor foci found in other organs , however; tumor emboLi were present in some of the vessels of the larger liver tumors. All liver lesions, observed by light microscopy, summarized in Table III. The other changes described in the liver and other tissues (Table IV) were regarded as lesions of naturally occurring diseases. The lesions were present, in most instances, among both control and test animals. The small (1-3 mm) , red, subcapsular foci observed at necropsy in the liver of both control and test animals could be correlated with scattered foci of hepatocellular necrosis observed histologically in these animals. Donovan E. Gordon, D.V.M., Ph.D. Diplomate, American College of Veterinary Pathologists jeuindtiiai B I O - T E 5 T Jialttyiabytied., 3etc. 7 R&S 135122 TABLE I TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats One-Year Interim Sacrifice Individual Gross Pathologic Findings in the Liver - Males Grouo Animal Number Gross Pathologic Findings Control 39 54 55 69 84 1 mm dark red foci in all lobes None Few 1 mm dark red foci in all lobes Numerous 1 mm dark red foci in all lobes None T-I 205 256 264 279 299 Few 1 mm dark red foci in all lobes 1-3 mm dark red foci in all lobes None None Numerous 1-3 dark red depressed foci in all lobes T-n T-m 404 433 459 469 489 609 639 663 668 694 Numerous 1-5 mm dark red depressed foci in all lobes Few 1-3 mm dark red foci in all lobes Few 1 mm dark red foci in all lobes 1 mm dark red foci in all lobes 1 mm dark red foci in all lobes, slight pale discoloration None Numerous 1 mm dark red foci in all lobes Granular surface with numerous 1 mm dark red foci in all lobes 1 mm dark red focus in 1 lobe 0.5 cm dark red subcapsular nodule in 1 lobe with capsular adhesions to spleen Dtidudi/iioL BIO-TEST ^aluyuittyue^, 9*ic, 8 R&S 135123 TABLE I continued TEST MATERIAL,: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats One-Year Interim Sacrifice Individual Gross Pathologic Findings in the Liver - Females Group Animal Number Gross Pathologic Findings . Control 106 149 160 178 200 1-3 mm dark red foci in all lobes Few 1 mm dark red foci in all lobes None Numerous 1 mm red foci in all lobes Few 1 mm red foci in all lobes T-I 303 349 369 381 397 Few 1 mm dark red foci in all lobes 1-3 mm dark red depressed foci in all lobes 1-3 mm dark red foci in all lobes 1-3 mm dark red foci in all lobes Few 1-2 mm dark red foci in all lobes T-II 524 550 569 594 599 Few 1-3 mm dark red foci in all lobes 0.5 cm dark red nodule in 1 lobe 1 mm dark red foci in all lobes 1-2 mm dark red foci in all lobes 4 cm subcapsular nodular focus in major lobe, capsular adhesions to diaphragm, spleen and serosal surface of small intestine T-III 717 728 732 735 742 0.5 cm dark red focus in 1 lobe with capsular adhesions to mesentery None 1 cm dark red focus in 1 lobe 3 cm nodule in 1 lobe; 0.5 cm dark red nodule in another lobe; focal capsular adhesions between nodule, small intestine, stomach, spleen and mesentery 4 mm dark red depressions in 1 lobe, 1-5 mm dark red foci in all other lobes b io - test a TABLE II TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats One-Year Interim Sacrifice Summary of Gross Pathologic Findings in All Tissues Except Liver Organ Lung Kidneys Ovaides Group: Number per Group: Findings Focal adhesions to diaphragm or thorax Few pin-point red foci in all lobes Focal consolidation of 1 or more lobes Abscesses in parenchyma Unilateral hydronephrosis Bilateral hydronephrosis Unilateral or bilateral follicular cysts Control 10/10 Male/Female Incidence T-I 10/10 Male/Female Incidence T-II 10/10 Male/Female Incidence T-II I 10/10 Male/Fcmale Incidence | ^ 3/0 1/0 2/0 0/0 1/0 0/0 0/0 1/0 0/1 0/0 0/1 I/O 0/0 1/0 0/0 0/0 0/0 1/0 0/0 0/0 1/0 0/0 0/0 0/0 220 1 Adrenal glands Small (1 mm) blood-filled cysts in cortex of 0/1 0/0 0/0 0/0 1 gland Enlarged and dark red color 0/0 0/0 0/1 0/0 Uterine horns Ilydrometra 2000 bZISZl SSH SZIQZl SSH 1 s 3 - O I 9i TABLE II continued TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats One-Year Interim Sacrifice Summary of Gross Pathologic Findings in All Tissues Except Liver Group: Control T-I T-II T-III Number per Group: 10/10___________ 10/10___________ 10/10____________ 10/10 , Male/Female Male/Female Male/Female Male/Female OrganFindingsIncidenceIncidenceIncidenceIncidence Pituitary gland Hemorrhagic focus 0/0 1/0 0/0 0/0 Spleen Few adhesions to mesentery Enlarged (3 times normal size) 0/0 0/0 1/0 1/0 0/0 0/0 0/1 0/0 Abdominal cavity Small amount of blood in peritoneal cavity 0/0 0/0 0/1 0/0 *6 0ual B I - T ES T Js&yM&yutd, $nc. TAIM.U 111 TEST MATKltlAL: Vinyl Chloride (Ethylene Derived) Chrunlc Vapor Inhalation Study ~ Albino flat* One-Year Interim Sacrifice Cj'OUp ami Auluietl Namlicr lillo Duel Proliferation Poila) lo Sinusoidal H&hamcUuMijry Llunialo\*i>Lv*jl:i Necruuili oi Kt-pjlocy tua Summary of Diver Lebiona* -Male* atomic llypuitrujtlty Lipid Vatcuolea of fit llepitocy l:l lly pcrlicijihy 4iui llypcrjilaiilii of lLc|ato<:ylm* Slnuuotdal Blood-Filled Dilatation________________________ Cyallc Subcapbiilar Thi-ouyliout Space* lien Din Oidy Section________________________________(Polios!*)Anulosarcum* Col i li'ol 39 51 55 19 Hi 1-2 2/M 1-2/M 2/M 1 1-2 T-l 205 269 279 299 T-Il 404 41J 959 Ifl 4dS i 1-2 1 1 1-2/M t (few) 1-2 1-2 i 1 1-2 l (few) 1 1-2 9/D (1 lobe) i 2l 2/M 1-2 2/M 2/M 2 1-2 1-2 1 1-2 1-2 1 \ 1-2 1 1-2 1-2 1 1-2 2 I T-l 11 Lu`i 639 tu t*lH2 I 1 1 1-2 1 t 2/M l 2 1-2/M 1 2 1 1 2/M 1-2/M 2/M 2-J/W 2/M I 2-) 1 2 1* P PP PP * All lesion* ate focal unices otherwise indicated. Gradmit S y slent ami AhiircvialDina 1 a minimal in beverily 2 - mild In beverily P a PrObeilt. nu ^rade 1 - moderate In bevel lly M * mulllfuCal 1 a mai ked ill b eve I'll y 11 a dlflube 5 a exit elite In veveilly gzvsei- s'su lzvsev s'sb BIO-TEST TABLE 111 cmitinued TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Kale Onc-Ycar Interim Sacrifice Suinuury ol Liver Leukine* * Female* Group and Animal Number UUe Duct Prolifer ution Furtal Lo Sinusoidal Extramedullary I tc mj lojiulcbly Necrosis of Llenalocvtcs Cytoplasmic Hypertrophy Lipid Vacuole* of lu 1 Icpalocylcs UepaiucyleS Hypertrophy and Hyperplasia of Hepalncylc* Sinusoidal Dilatation Subcapsular Throughout It epical Only Section Control IOC ICO r/a 2U0 i 1-2/M 2 il i i i i T-1 30 J 3-tV 3C`J id 1 1V7 i 1 2 (few) i i i i 1-2 1-2 1-2 1-2 1 Til u\ SiO it'} S-H W i i i i i 1-2 1-2 1 I 1-2 1-2/M 2/M i 2 1-1 2 T- 111 717 72d 712 7 IS 7-12 i i i 1-3 1 1 1-2 1 2 t 1-2 1 2/M 2-1/M 2-1/M 2-1/M 2/M 2 1-2 2 2 2 Blood- Filled Cystic SpaCe* (Pullunla) Anulosurcoma V 1 Pp Pp Pp Pp Pp Pp * All legion* are focal uriluSM otherwise Indicated. Orjtlm[j Sysliin *inl Abbreviations 1 - minimal In buvcrlty 2 - mild in severity 3 - uiucliTJlu hi icvtiily 1* * Present, no yrade !! = multifocal 4 - maiVed in severity 1) - diffuse In seventy W B10-THST I* TABLE IV TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Hula One-Year Interim Sacrifice Incidental Histopathologic Changes In Tisaueu Other Than Liver - Maluti Orjpn LJn dint:* Gvoup: Number Examined: Cunlrol 10 Average incidence Grade T-l 10 Average Incidence Grade t-h 10 Average Incidence Grade T--III 10 Average Incidence Grade Trachea Chronic traehcllla 2 2.0 3 1.0 2 i.a 3 1.7 Lung Chronic focal murine pneumonia Local Aggregates of alveolar macrophages Acute focal bronchopneumonia 3 1.3 2 1.0 5 1.3 1 2.0 2 1.5 1 1.0 5 1.3 2 1.0 Heart Local myocardial degeneration and fibrosis 2 1.0 2 1.0 1 1.0 Kidney Bilateral focal prolcinaceouu tubular casts Bilateral local tubular nephrosis UUalera) focal Inlcrsiltiat lymphoid infiltrations Bilateral chronic focal nephritis Bilateral focal minera )i <tcd jii icruconcretions in tub iuTl* Unilateral hydronephrosis Bj lateral liydronephrosis 3 1 2 1 1.2 1.5 1.0 2.5 2 1.0 1 1.0 1 1.5 1 2.0 2 i.a 2 1.8 1 1.0 4 1.5 3 1.7 1 1.0 Urinary bladder LrotcknaCeuus ploy In lumen I'rostulu gland Local in Jet a t Ilia 1 lymphoid infiltrations 2 1.0 1 1.0 3 1.0 1 1.0 4 1.0 Tea Icy Unilateral focal teslicular degeneration Unilateral focal lenticular degeneration with spermatoceles * spcrmalie granulemas and dyslruphic minerdlixaliuu of some affected tubules 1 1.0 1 3.5 831-901. S?U U U toel BIO -TEST 621-SeV SSti TABLE IV continued TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rats Oac'Ycar Interim Sacrifice Incidental llistopalhologic Changes In Tissues Other Than Liver " Males Organ Piluitary I'ldlul Adrenal glands Findin gs Group: Mimiljcr Examined: Control 10 Average lucid ence Grade T-l 10 Average Incidence Grade T-1I 10 Average lncldence Grade T-m 10 Average Incidence Grade Focal cytoplasmic vacuolailon of basophils Small chromophobe adenoma 2 1.0 2 1.0 1P 1 1.0 Unilateral focus of lipomatous degeneration in cortex 2 1.0 1 l.S 1 1.0 Thy rojd gland* Bilateral diffuse hypertrophy of follicular epithelial Cells 1 2.0 Colon Nematode purabltea In lumen (jiciuulodtasis) 1 - Bone Focal medial hypertrophy ol arterioles in femoral marrow 1 1.0 1 2.0 1 1.0 1 2.0 Grading System 1.0 - minima) In severity 2.0 - mild in Severity 3.0 - moderate in bcvurtty 4.0 = marked in severity 5.U - extreme In severity l* = Present, no grade BIO-TEST Organ Tracliea Lung Heart Kidneys Spleen U lei* us TABLE IV continued TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Rate One-Ycur Interim Sacrifice Incidental 1 ilalop jllmlo^ Ic Changes in Tlueuea Other Than Liver - Font a lea Findings Group; Number Examined; Con trul 10 Average Incidence Grade T-I 10 Avurauu Incidence Grade T-II 10 Average Incidence Grade T-m 10 Average Incidence Grade Chronic tracheitis 3 1.8 3 1.5 2 1.8 1 1.0 Chronic focal murine pneumonia Focal ayti resales of alveolar macrophages Diffuse hemorrhage and edema (agonal) Focal suppurative bronchiolitis 5 1.2 2 1.0 1 3.0 1 1.0 4 1.0 4 1.8 1 2.0 4 1.1 1 1.0 Few small foci of chronic myocarditis 1 1.0 Unilateral focal proteinaceous luhular casta bilateral focal proteinaceous tubular casts bilateral local tubular nephron is Uuilalcral chronic focal nephritis Unilateral focal mineralized micructmcrelJons in tubules bilateral focal mineralized microcuncretIons in tubules Unilateral targe tubular cyst in cortex bilateial diffuse brown granular pigment and protein resorption droplets within cytoplasm i)t cortical tubular epithelial Cells i 1 1 1 4 1.0 2.0 l.S 1.0 1.') 3 1.0 1 1.0 3 2.0 1 1.0 4 1.0 1 1.5 l 1.0 1 1.0 4 l.S 1 t* Increased hemosiderin pigment Diffuse increased cxlramedu It a ry hematopoiesis llydvumclra Suppurative metritis with endometrial abscesses Suppurative metritis 2 1 1.5 3.0 2 1.5 1 1.0 1 3.0 1 2.0 2 1.0 1 2.0 1 2.0 oe wse u ssu tet.se i. ssu BIO -TEST 1* TAtiLE IV continued TEST MATERIAL: Vinyl Chloride (Ethylene Derived) Chronic Vapor Inhalation Study - Albino Hats One-Year Interim Sacrifice Incidental Histopathologic Changed in Tissues Other Than Liver - Females Ori*an Ovaries Findings Group; Humber Examined: Cun trot 10 Average Incidence Grade T-l 10 Average Incidence Grade T-H 10 Average Incidence Grade T-11I 10 Average Incidence Grade bilateral local follicular cysts Unilateral focal follicular cysts ip ip i l* 2P Pituitary ytand Focal hyperplasia of chromophobe cells 2P Adrenal glands Unilateral focus of llpomatuus degeneration in cortex Unilateral focal hemalocysts in cortex bilateral focal hematucysls In cortex 1 4.0 i 1.0 1 2.0 2 2.0 1 1.0 1 3.0 1 2.0 Pancreas Thyroid gland focal aliojiliy of acinar cells bilateral diffuse hypertrophy of fullicular eplthelial cells 1 1.0 1 1.0 9 1 2.0 1 1.0 Uyc Unilateral clironlc focal bacryoudenilis Unilateral focal retinal degeneration 1 1.0 i 1.0 Colon Nematode parasites in lumen (ncmaludiasls) 2P Orailmy System 1.0 = minimal In severity 2.0 = mild in severity 3.0 = moderate in severity 4.0 = mutUud in severity 5.0 = extreme i11 severity F = I1 resent, no grade