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資料6
出典 National Cancer Institute, TR-97. Bioassay of Titanium Dioxide for Possible
U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFAREPublic Health ServiceNational Institutes of Health
BIOASSAY OF
TITANIUM DIOXIDE
FOR POSSIBLE CARCINOGENICITY
Carcinogenesis Testing ProgramDivision of Cancer Cause and Prevention
National Cancer InstituteNational Institutes of Health
Bethesda, Maryland 20014
U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFAREPublic Health Service
National Institutes of Health
DREW Publication No. (NIH) 79-1347
ii
BIOASSAY OFTITANIUM DIOXIDE
FOR POSSIBLE CARCINOGENICITY
Carcinogenesis Testing ProgramDivision of Cancer Cause and Prevention
National Cancer InstituteNational Institutes of Health
FOREWORD; This report presents the results of the bioassay oftitanium dioxide conducted for the Carcinogenesis TestingProgram, Division of Cancer Cause and Prevention, National CancerInstitute (NCI), National Institutes of Health, Bethesda,Maryland. This is one of a series of experiments designed todetermine whether selected chemicals have the capacity to producecancer in animals. Negative results, in which the test animalsdo not have a greater incidence of cancer than control animals,do not necessarily mean that the test chemical is not acarcinogen, inasmuch as the experiments are conducted under alimited set of circumstances. Positive results demonstrate thatthe test chemical is carcinogenic for animals under theconditions of the test and indicate that exposure to the chemicalis a potential risk to man. The actual determination of the riskto man from animal carcinogens requires a wider analysis.
CONTRIBUTORS; This bioassay of titanium dioxide was conducted byHazleton Laboratories America, Inc., Vienna, Virginia, initiallyunder direct contract to NCI and currently under a subcontract toTracer Jitco, Inc., prime contractor for the NCI CarcinogenesisTesting Program.
The NCI project officers who were responsible for selecting theprotocols used in this bioassay were Drs. N. P Page^'^ and C.Cuetol. The principal investigators were Drs. M. B. Powers^ andR. W. Voelker^. Ms. K. J. Petrovics^ was responsible for datamanagement, and Mr. G. Najarian^ for animal care.Histopathologic examinations were performed by Drs. D. A. Banas-*and R. H. Habermann^ and reviewed by Dr. Voelker, and thediagnoses included in this report represent their interpretation.
iii
Animal pathology tables and survival tables were compiled at EG&GMason Research Institute^. The statistical analyses wereperformed by Dr. J. R. Joiner5 and Ms. P. L. Yong5, using methodsselected for the bioassay program by Dr. J. J. Gart6. Chemicalsused in this bioassay were analyzed at Midwest Research Instituteunder the direction of Dr. E. Murrill^ and feed mixturescontaining the test chemical were analyzed at HazletonLaboratories by Dr. C. L. Guyton^ and Mr. E. Missaghi^. xheresults of these analyses were reviewed by Dr. S. S. Olin5.
This report was prepared at Tracer Jitco5 in collaboration withHazleton Laboratories and NCI. Those responsible for the reportat Tracer Jitco were Dr. L. A. Campbell, Director of the BioassayProgram; Dr. S. S. Olin, Deputy Director for Science; Dr. J. F.Robens, toxicologist; Dr. R. L. Schueler, pathologist; Dr. G. L.Miller, Ms. L. A. Waitz, and Mr. W. D. Reichardt, biosciencewriters; and Dr. E. W. Gunberg, technical editor, assisted by Ms.Y. E. Presley and Ms. P. J. Graboske.
The following other scientists at NCI were responsible forevaluating the bioassay experiment, interpreting the results, andreporting the findings: Dr. Kenneth C. Chu, Dr. Cipriano Cueto,Jr., Dr. J. Fielding Douglas, Dr. Dawn G. Goodman**, Dr. RichardA. Griesemer, Dr. Morton H. Levitt, Dr. Harry A. Milman, Dr.Thomas W. Orme, Dr. Robert A. Squire9, Dr. Sherman Stinson, Dr.Jerrold M. Ward, and Dr. Carrie E. Whitmire.
^Carcinogenesis Testing Program, Division of Cancer Cause andPrevention, National Cancer Institute, National Institutes ofHealth, Bethesda, Maryland.
2Now with the Environmental Protection Agency, 401 M Street,S.W., Washington, D.C.
Mason Research Institute, 1530 East Jefferson Street,Rockville, Maryland.
IV
^Tracer Jitco, Inc., 1776 East Jefferson Street, Rockville,Maryland.
^Mathematical Statistics and Applied Mathematics Section,Biometry Branch, Field Studies and Statistics, Division ofCancer Cause and Prevention, National Cancer Institute,National Institutes of Health, Bethesda, Maryland.
^Midwest Research Institute, 425 Volker Boulevard, Kansas City,Missouri.
with the Division of Comparative Medicine, Johns HopkinsUniversity, School of Medicine, Traylor Building, Baltimore,Maryland.
vi
SUMMARY
A bioassay of titanium dioxide for possible carcinogenicity wasconducted by administering the test chemical in feed to Fischer 344rats and B6C3F1 mice.
Groups of 50 rats of each sex and 50 mice of each sex wereadministered titanium dioxide in the diet at one of two doses, either25,000 or 50,000 ppm, for 103 weeks and then observed for 1additional week. Matched controls consisted of 50 untreated rats ofeach sex and 50 untreated mice of each sex. All surviving rats andmice were killed at 104 weeks.
Administration of the titanium dioxide had no appreciable effect onthe mean body weights of rats or mice of either sex. With theexception of white feces, there was no other clinical sign that wasjudged to be related to the administration of titanium dioxide.Survival of the rats and the male mice at the end of the bioassay wasnot affected by the test chemical; mortality in female mice was doserelated. Sufficient numbers of dosed and control rats and mice ofeach sex were at risk for development of late-appearing tumors.
In the female rats, C-cell adenomas or carcinomas of the thyroidoccurred at incidences that were dose related (P = 0.013), but werenot high enough (P = 0.043 for direct comparison of the high-dosegroup with the control group) to meet the level of P = 0.025 requiredby the Bonferroni criterion (controls 1/48, low- dose 0/47,high-dose 6/44). Thus, these tumors of the thyroid were notconsidered to be related to the administration of the test chemical.
In the male and female mice, no tumors occurred in dosed groups atincidences that were significantly higher than those forcorresponding control groups.
It is concluded that under the conditions of this bioassay, titaniumdioxide was not carcinogenic by the oral route for Fischer 344 ratsor B6C3F1 mice.
vii
viii
TABLE OF CONTENTS
Page
I. Introduction 1
II. Materials and Methods 3
A. Chemical 3B. Dietary Preparation 4C. Animals 5D. Animal Maintenance 5E. Subchronic Studies 8F. Designs of Chronic Studies 9G. Clinical and Pathologic Examinations 9H. Data Recording and Statistical Analyses 12
III. Results - Rats 17
A. Body Weights and Clinical Signs (Rats) 17B. Survival (Rats) 17C. Pathology (Rats) 20D. Statistical Analyses of Results (Rats) 21
IV. Results - Mice 25
A. Body Weights and Clinical Signs (Mice) 25B. Survival (Mice) 25C. Pathology (Mice) 28D. Statistical Analyses of Results (Mice) 29
V. Discussion • 31
VI. Bibliography 35
APPENDIXES
Appendix A Summary of the Incidence of Neoplasms inRats Administered Titanium Dioxide in the Diet 39
Table Al Summary of the Incidence of Neoplasms in Male RatsAdministered Titanium Dioxide in the Diet 41
Table A2 Summary of the Incidence of Neoplasms in Female RatsAdministered Titanium Dioxide in the Diet 45
ix
Page
Appendix B Summary of the Incidence of Neoplasms in MiceAdministered Titanium Dioxide in the Diet..... 49
Table Bl Summary of the Incidence of Neoplasms in Male MiceAdministered Titanium Dioxide in the Diet..... 51
Table B2 Summary of the Incidence of Neoplasms in FemaleMice Administered Titanium Dioxide in the Diet 54
Appendix C Summary of the Incidence of NonneoplasticLesions in Rats Administered Titanium Dioxidein the Diet 59
Table Cl Summary of the Incidence of NonneoplasticLesions in Male Rats Administered Titanium Dioxidein the Diet 61
Table C2 Summary of the Incidence of NonneoplasticLesions in Female Rats Administered Titanium Dioxidein the Diet 67
Appendix D Summary of the Incidence of NonneoplasticLesions in Mice Administered Titanium Dioxidein the Diet 73
Table Dl Summary of the Incidence of NonneoplasticLesions in Male Mice Administered Titanium Dioxidein the Diet 75
Table D2 Summary of the Incidence of NonneoplasticLesions in Female Mice Administered Titanium Dioxidein the Diet 79
Appendix E Analyses of the Incidence of Primary Tumorsin Rats Administered Titanium Dioxide in the Diet.. 83
Table El Analyses of the Incidence of Primary Tumors in MaleRats Administered Titanium Dioxide in the Diet 85
Table E2 Analyses of the Incidence of Primary Tumors in FemaleRats Administered Titanium Dioxide in the Diet 92
Appendix F Analyses of the Incidence of Primary Tumorsin Mice Administered Titanium Dioxide in the Diet.. 99
x
Pace
Table Fl Analyses of the Incidence of Primary Tumors in MaleMice Administered Titanium Dioxide in the Diet 101
Table F2 Analyses of the Incidence of Primary Tumors in FemaleMice Administered Titanium Dioxide in the Diet..... 104
Appendix G Analysis of Formulated Diets forConcentrations of Titanium Dioxide 109
TABLES
Table 1 Design of Titanium Dioxide Chronic FeedingStudies in Rats and Mice 10
FIGURES
Figure 1 Growth Curves for Rats Administered TitaniumDioxide in the Diet 18
Figure 2 Survival Curves for Rats Administered TitaniumDioxide in the Diet 19
Figure 3 Growth Curves for Mice Administered TitaniumDioxide in the Diet 26
Figure 4 Survival Curves for Mice Administered TitaniumDioxide in the Diet 27
xi
xii
I. INTRODUCTION
Titanium dioxide (CAS 13463-67-7; NCI C04240) is a white pigment
possessing great covering or opacifying power. It exists in
three crystalline forms: anatase, brookite, and rutile, but only
the anatase variety is used as a food color additive (Noonan,
1975). Titanium dioxide has been in use since 1918, although the
market was greatly expanded after 1948 when the need for titanium
led to technological advancements in ore processing (Bomberger,
1969). In 1977, the production volume for titanium dioxide in
the United States was 800,000 tons. The majority of this was
produced for pigmentary applications; 50% for paints and other
protective coatings, 20% for paper, and 12% for plastics (Greek,
1977). Titanium dioxide is used as a color additive in foods
(anatase) (FDA, 1976a), and in topical and oral drugs (FDA,
1976b). In the cosmetics industry, it is used as a whitener in a
wide variety of products including aftershave powders, bath
powders, face powders, depilatories, deodorants, fingernail
Inc., Des Moines, Iowa). The sodium pentobarbital was injected
intraperitoneally at a volume of 0.3 to 0.5 ml for the rats and
0.03 to 0.05 ml for the mice.
The pathologic evaluation consisted of gross and microscopic
examination of major tissues, major organs, and all gross lesions
Table 1. Design of Titanium Dioxide Chronic Feeding Studiesin Rats and Mice
Sex andTestGroup
Male
Matched-Control
Low -Dose
High-Dose
Female
Matched-Control
Low-Dose
High-Dose
InitialNo. ofAnimals3
50
50
50
50
50
50
TitaniumDioxideDosesb
(ppm)
0
25,000
50,000
0
25,000
50,000
TimeDosed(weeks)
103
103
103
103
on StudyObserved(weeks)
104
1
1
104
1
1
aRats were 64 days of age and mice were 36 days of age when placedon study.
test chemical was administered 7 days per week in a dietcontaining 2% corn oil. The control groups received only2% corn oil in the diet. Diets were available ad libitum.
10
from killed animals and from animals found dead. The tissues
were preserved in 10% buffered formalin, embedded in paraffin,
sectioned, and stained with hematoxylin and eosin. The following
tissues were examined microscopically: brain (frontal cortex and
basal ganglia, parietal cortex and thalamus, and cerebellum and
pons), pituitary, spinal cord (if neurologic signs were present),
irritation and swelling of the testes, hunched appearance, and/or
thinness. Alopecia (localized or generalized) was noted in all
the control and dosed groups; however, more was observed in the
control females than in the dosed females. The areas of alopecia
were primarily located around the nose and head and progressed to
generalized alopecia in some of the animals. The type of feed-
hopper used in this study may have caused the^alopecia around the
nose. Animals in all of the dosed groups had white feces.
B. Survival (Mice)
The Kaplan and Meier curves estimating the probabilities of
survival for male and female mice administered titanium dioxide
in the diet at the doses of this bioassay, together with those of/'
the matched controls, are shown in figure 4. In male mice, the
result of the Tarone test for dose-related trend in mortality is
25
30 HUJgoto
< 20 -
e a n n
MALE MICED MATCHED CONTROL
O LOW DOSE
A HIGH DOSE
10 20 30 40 50 60 70
TIME ON STUDY (WEEKS)
100 110
_ 35(9
>- 25QOmZ
2A A
Aa
An a D a
aA
a a
FEMALE MICED MATCHED CONTROL
O LOW DOSE
A HIGH DOSE
100 110
TIME ON STUDY (WEEKS)
Figure 3. Growth Curves for Mice Administered Titanium Dioxide in the Diet
26
_J
>
(Au.
>Hj
CD0cc
u
M
D MAI
O LOV
A HIG
\-l^
\LE Ml
FCHED CO
IDOSE
H DOSE
CE
-J, ti~~&
Qi, c
A..
HV
^H
^
"Ab — &-•
5 i
7,
(
LW^1 -
U
0 10 20 30 40 50 60 70 80 90 100
TIME ON STUDY (WEEKS)
<>
tou.O
HZima0ccCL
A-
FEI\
a MAO LOV
A Hie
1ALE M
rcHEDCO
(DOSE
HDOSE
ICE
T ) — —
i
~~b-
i
6—
"•\..
' 1•J.
*
A^
^0
^i.
\
10 20 30 40 50 60 70 80 90 100 110
TIME ON STUDY (WEEKS)
Figure 4. Survival Curves for Mice Administered Titanium Dioxide in the Diet
27
not significant, but in females, the result of the Tarone test
shows a significant (P = 0.001) positive dose-related trend.
Forty out of fifty (80%) of the high-dose males, 40/50 (80%) of
the low-dose males, and 32/50 (64%) of the matched-control males
were still alive at week 104. In females, 33/50 (66%) of the
high-dose group, 39/50 (78%) of the low-dose group, and 45/50
(90%) of the matched controls were alive at week 104. Sufficient
numbers of mice of each sex were at risk for the development of
late-appearing tumors.
C« Pathology (Mice)
Histopathologic findings on neoplasms in mice are summarized in
Appendix B, tables Bl and B2; findings on nonneoplastic lesions
are summarized in Appendix D, tables Dl and D2.
A low incidence of neoplasia was observed in both the control
mice and dosed mice. These neoplasms were of the usual number
and type observed in mice of this age and strain. A slightly
increased number of hepatocellular carcinomas was observed in the
high-dose males; however, the incidence of tumors was not
increased over that observed in historical-control groups of mice
of this age and strain.
28
Degenerative, proliferative, and inflammatory lesions were also
of the usual number and kind observed in aged B6C3F1 mice.
Based on the histopathologic examination, titanium dioxide was
neither toxic nor carcinogenic to B6C3F1 mice under the
conditions of this bioassay.
D. Statistical Analyses of Results (Mice)
Tables Fl and F2 in Appendix F contain the statistical analyses
of the incidences of those primary tumors that occurred in at
least two animals of one group and at an incidence of at least 5%
in one or more than one group.
The results of the Cochran-Armitage test for positive dose-
related trend in incidences of tumors and those of the Fisher
exact test for higher incidences of tumors in dosed groups than
in control groups are not significant for any type of tumor
occurring in either sex. A significant trend (P = 0.037) in the
negative direction is observed in the incidence of follicular-
cell adenomas of the thyroid in female mice, in which the
incidence in the control group exceeds the incidences in the
dosed groups. The results of the Fisher exact test (P = 0.035 in
the negative direction) for the comparison of the incidence of
combined lymphomas and leukemias in the female low-dose group
with that in the corresponding controls are above that of 0.025
29
required for significance in multiple comparisons. This negative
result may be accounted for by the difference in survival, since
the dosed animals did not live as long as the control animals.
In each of the 95% confidence intervals of relative risk, shown
in the tables, the value of one is included; this indicates the
absence of significant positive results. It should also be noted
that each of the intervals has an upper limit greater than one,
indicating the theoretical possibility of the induction of tumors
by titanium dioxide, which could not be detected under the
conditions of this test.
30
DISCUSSION
Based on growth rate, mortality, and other clinical signs, there
was essentially no evidence of toxicity of titanium dioxide in
the dosed rats or dosed mice. Administration of the test
chemical had no appreciable effect on the mean body weights of
either male or female rats with the exception of white feces,
there was no other clinical sign that was judged to be related to
the administration of titanium dioxide. Survival of the male and
female rats and of the male mice at the end of the bioassay was
not affected by the test chemical; survival of the high-dose
female mice was shorter than that of the low-dose and control
groups. Sufficient numbers of dosed and control rats and mice of
each sex were at risk for development of late-appearing tumors.
Although little or no effect on weight gain and survival could be
attributed to titanium dioxide, except in female mice, the doses
were considered to approximate the maximum that could be
administered and still not affect the nutritional quality of the
diet. This is consistent with the guidelines for carcinogenesis
bioassay in the Carcinogenesis Testing Program (Sontag et al.,
1976).
In the female rats, C-cell adenomas or carcinomas of the thyroid
occurred at incidences that were dose related (P = 0.013), but
not high enough (P = 0.043 for direct comparison of the high-dose
31
group with the control group) to meet the level of P = 0.025
required by the Bonferroni criterion (controls 1/48, low-dose
0/47, high-dose 6/44). Thus, the tumors of the thyroid are not
considered to be related to administration of the test chemical.
Also in the females, endometrial stromal polyps of the
endometrium/uterus occurred at. higher incidences in the dosed
groups than in the controls, but the incidences were not dose
related and were not high enough (P = 0.045 for direct comparison
of the low-dose group with the control group) to meet the
requirements of the Bonferroni criterion (controls 7/50, low-dose
15/50, high-dose 10/50),
In the male and female mice, no tumors occurred in dosed groups
at incidences that were significantly higher than those in
corresponding control groups.
In other studies, no adverse pulmonary effects were foun4' when
Wistar rats were administered titanium dioxide by inhalation
(Christie et al., 1963), and no evidence of carcinogenicity was
found when Swiss albino mice were administered potassium titanium
oxalate at a concentration of 5 ppm titanium in drinking water
for the life span of the mice (Schroeder et al. , 1964). When
titanium was administered to Fischer 344 rats and to DBA/2,
C57BL/6, or Swiss albino mice by intramuscular injection as
titanocene, a complex of titanium with cyclopentadiene, a variety
32
of neoplasms developed at the site of injection and in organs
some distance away (Furst and Haro, 1969, 1970).
It is concluded that under the conditions of this bioassay,
titanium dioxide was not carcinogenic for Fischer 344 rats or
B6C3F1 mice.
33
34
VI. BIBLIOGRAPHY
Armitage, P., Statistical Methods in Medical Research, John Wiley& Sons, Inc., New York, 1971, pp. 362-365.
Association of Official Analytical Chemists, Official Methods ofAnalysis _of the Asociation of Official Analytical Chemists,12th edition, Horwitz, W., ed., Association of OfficalAnalytical Chemists, Washington, B.C., 1975, p. 7.
Barry, R. H., Depilatories. In: Cosmetics - Science andTechnology, Vol. _2, Balsam, M. S. and Sagarin, E. , eds., NewYork, 1972, pp. 49 and 57.
Bell, S. A., Preshave and aftershave preparations. In:Cosmetics - Science and Technology, Vol. _2_, Balsam, M. S.and Sagarin, E., eds., Wiley-Interscience, New York, 1972,p. 33.
Berenblum, I. , ed., Carcinogenicity Testing, A_ Report of thePanel on Carcinogenicity of the Cancer Research Commissionof the UICC, Vol. _2, International Union Against Cancer,Geneva, 1969.
Bomberger, H. B., Titanium and titanium alloys. In: Kirk - OthmerEncyclopedia £f Chemical Technology, Vol. 20, Mark, H. F. ,McKetta, J. J. , Jr., and Othmer, D. F., eds., IntersciencePublishers, Inc., New York, 1969, pp. 347-379.
Christie, H. , MacKay, R. J. , and Fisher, A. M., Pulmonary effectsof inhalation of titanium dioxide by rats. Amer. Ind. Hyg.ASSOC. J. 24:42-46, 1963.
Cox, D. R. , Regression models and life tables. J. R. Statist.Soc. B 34:187-220. 1972.
Cox, D. R. , Analysis of Binary Data, Methuen & Co., Ltd., London,1970, pp. 48-52.
Doviak, W. C., Nail lacquers and removers. In: Cosmetics -Science and Technology, Vol. _2, Balsam, M. S. and Sagarin,E., eds., Wiley-Interscience, New York, 1972, pp. 527-529.
Farber, L., Face powders. In: Cosmetics - Science and Technology,Vol. _1, Balsam, M. S. and Sagarin, E. , eds., Wiley-Interscience, New York, 1972, pp. 338-339.
35
Fielder, J. G. , Foundation makeup. In: Cosmetics - Science andTechnology, Vol. _1, Balsam, M. S. and Sagarin, E., eds.,Wiley-Interscience, New York, 1972, pp. 317-318.
Food and Drug Administration, 21 CFR 8.316, 1976a.
Food and Drug Administration, 21 CFR 8.6005, 1976b.
Furst, A. and Haro, R. 1., Carcinogenicity of metal pi-complexcompounds: metallocenes. In: Tenth International CancerCongress - Abstracts, Houston, Tex., May 22-29, 1970, p. 28.
Furst A. and Haro, R. T. , A survey of metal carcinogenesis.Progr. Exp. Tumor Res. 12:102-133, 1969.
Gart, J. J., The comparison of proportions: a review ofsignificance tests, confidence limits and adjustments forstratification. Rev. Int. Statist. Inst. 39(2);148-169,1971.
Greek, B. F., Two major pigments move into better times. C & EN_55 (24): 10-11, 1977.
Kammori, 0., Yamaguchi, N., and Sato, K. , Application of infraredabsorption spectrum to studies on iron and steel. I.Infrared absorption spectra of metal oxides, Bunseki Kagaku,16:1050-1955, 1967.
Kaplan, E. L. and Meier, P., Nonparametric estimation fromincomplete observations. J. Amer. Statist. Assoc._5_3:457-481, 1958.
Lauffer, P. G. I., Lipsticks. In: Cosmetics - Science andTechnology, Vol. _!_, Balsam, M. S. and Sagarin, E., eds.,Wiley-Interscience, New York, 1972, p. 370.
Linhart, M. S., Cooper, J. A., Martin, R. L., Page, N. P., andPeters, J. A., Carcinogenesis bioassay data system. Comp.and Biomed. Res. 7:230-248, 1974.
MacLeod, T. M. and Frain-Bell, W., A study of physical lightscreening agents. Brit. J. Dermat. 92:149-156, 1975.
Miller, R. G., Jr., Simultaneous Statistical Inference,McGraw-Hill Book Co., New York, 1966, pp. 6-10.
36
Noonan, J., Color additives in food. In: Handbook of FoodAdditives, Second Edition, Furia, T. E., ed., CRC Press,Cleveland, Ohio, 1975, pp. 603-604.
Plechner, S. , Antiperspirants and deodorants. In: Cosmetics -Science and Technology, Vol. _2, Balsam, M. S. and Sagarin,E., eds., Wiley-Interscience, New York, 1972, p. 388.
Saffiotti, U. , Montesano, R. , Sellakumar, A. R., Cefis, F., andKaufman, D. G., Respiratory tract carcinogenesis in hamstersinduced by different numbers of administrations of benzo (a)pyrene and ferric oxide. Cancer Res. 32:1073-1081, 1972.
Saute, R. E. , Bath preparations. In: Cosmetics - Science andTechnology, Vol. _2_, Balsam, M. S. and Sagarin, E., eds. ,Wiley-Interscience, New York, 1972, p. 514.
Schroeder, H. A., Balassa, J. J., and Vinton, W. H., Jr.,Chromium, lead, cadmium, nickel, and titanium in mice:effect on mortality, tumors and tissue levels. J. Nutritionj$3_:239-250, 1964.
Shevlin, E. J. , Skin lighteners and bleach creams. In:Cosmetics - Science and Technology, Vol. _1_, Balsam, M. S.and Sagarin, E. , eds., Wiley-Interscience, New York, 1972,p. 227.
Sontag, J. M., Page, N. P., and Saffiotti, V., Guidelines forCarcinogen Bioassay in Small Rodents. CarcinogenesisProgram, Division of Cancer Cause and Prevention, NationalCancer Institute, Bethesda, Md., 1976.
Stanley, R. H. , Titanium compounds (inorganic). In: Kirk - OthmerEncyclopedia _of_ Chemical Technology, Vol. 20, Mark, H. F.,McKetta, J. J. , Jr., and Othmer, D. F., eds., IntersciencePublishers, Inc., New York, 1969, pp. 347-379.
Tarone, R. E. , Tests for trend in life table analysis. Biometrikaj>2(3):679-682, 1975.
Wetterhahn, J., Eye makeup. In: Cosmetics - Science andTechnology, Vol. J_, Balsam, M. S. and Sagarin, E., eds.,Wiley-Interscience, New York, 1972, p. 397.
37
38
APPENDIX A
SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
RATS ADMINISTERED TITANIUM DIOXIDE IN THE DIET
39
40
TABLE A1.SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS
ADMINISTERED TITANIUM DIOXIDE IN THE DIET
1(
A N I M A L S I N I T I A L L Y I N STUDYA N I M A L S NECROrSISDA N I M A L S E X A M I N E D HI STOPATHOLOGIC ALL Y
I N T E G U M E N T A R Y SYSTEM
* S K I NS Q U A M O U S CELL P A P I L L O M AS Q U A M O U S CELL C A R C I N O M ABASAL-CELi, C A R C I N O M AK E R A T O A C A d T H O B A
*SUBCUT TISSJESQUAMOUS CELL PAPILLOMASQUAMOUS CELL CARCINOMABASAL-CELL C A R C I N O M AS A R C O M A , w O SFIBROMAFIBROSARGJMAL I P O M AH E M A N G I O S A R C O M AH E M A N G I O P E R I C Y T O M A , M A L I G N A N T
R 3 S P I R A T O R Y SYSTEM
*LUNGHEPATOCELiULAfi CARCINOMA, BETASTH E B A N G I O P E 8 I C Y T O M A , BETASTATIC
HEBATOPOIETIC SYSTEM
*MU1TIPLE O R G A N SGRANOLOCYJIC LEUKEBIABOHOCYTIC LEUKEHIA
ISP1EENHEMANGIOSARCOHA
WATCHEDSOIMTROL LOW DOSE
50 5049 5049 50
(49) (50)1 ( 2 % )
1 (2%)1 (2%)
(49) (50)1 (2%)
1 (2%)1 (2%)
1 (2%) 5 (10%)1 (2%) 2 (4%)
1 (2%)1 (2%)
1 (255)
(49) (50)1 (2*)
1 (2*)
(49) (50)2 (4X)
14 (29S) f~ ,-•(, (12*)
(49) (50)1 1281
HIGH DOSE
505050
(50)
2 (14%)
3 (6%)
(50)
1 (2%)5 (1055)
(49)
J (50)1 J2S)5 (10K)
(50)3_I£S1 _
t NUMBER OF AfllBALS WITH TISSDE EXAMINED HICROSCOPICALLY* N U M B E R OF AUIMALS NECHOPSIED
41
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)
MATCHEDCONTROL LOW DOSE HIGH DOSE
t T H Y M U S (48) (45) ( 2 8 )C A R C I N O M A , N O S 1 (2%)H E P A T O C E L i . U L A R C A R C I N O M A , BETAS'!' 1 (255)
C I R C U L A T O R Y S Y S T E M
i iONE
DIGESTIVE SYSTEM
K L I V E B ( 4 9 ) (50) (50)N E O P L A S T I C N O D U L E 1 (2%)H E P A T O C E L n U L A R C A R C I N O M A 1 (2%)HEM A N G I O S i i R C O M A , MET A S T A T I C 1 (2%)
*CECUfi (49) (46) (48)FIEROSARCoHA 1 (2%)
URINARY SYSTE.1
# K I D N E Y ( 4 9 ) (50) (50)M I X E D T U H v j R , B E N I G N 1 (2?>)
K U R I N A R Y B L A J D E R (48) (42) ( 4 5 )T R A N S I T I O . i A L - C E L L P A P I L L O M A 1 (2*)
E N D O C R I N E S Y S T E M
tPITUITARYCHROMOPHOdE AD3NOMA
*ADEENALP H E O C H R O H u C Y T O M A
• T H Y R O I DF O L L I C U L A h - C E L L A D E N O M AF O L L I C Q L A i - J - C E L L C A R C I N O M AC-CELL A D E N O M AC - C E L L C A r i C I N O M A
ffPANCSEAIIC ISLETS
(48)5 (1055)
( 4 9 )7 (14S)
(49)
1 (255)
4 (8%)
(50)10 (20%)
(49)9 ( 1 8 % )
(49)
1 (2%)3 (6%)1 (2%)
(50)
(46 )7 (15S)
(50)14 (28%)
(50)1 (2%)1 (2-7.)
1 <2X )
(50 )
* NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY* NUHBE3 OF ANIMALS NECROPSIES
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)
MATCHEDCONTROL LOW DOSE HIGH DOSE
1 (2X)ISLET-CELL C A R C I N O M A
REPRODUCTIVE SYSTEM
*MAMMARY GLAsIDFIBHOADENOMA
*PREPOTIAL GLANDCARCINOMA, NOS
(49)
1 (2%)
(49)
2 (4S)
tTESTIS (49)I N T E R S T I T i A L - C E L L TOH08 44 (90%)INTERSTITIAL-CELL T U M O R , M A L I G N A 1 (2X)
(50)1 (2%)
(50)5 (10%)
(49)46 (94%)
(50)3 (635)
(50)6 (12%)
(50)41 (82S)
*£PIDIDYMIS (49) (50) (50)I N T E R S T I T i A L - C E L L T U M O R , I N V A S I V 1 (2%)
NERVOUS SYSTErf
# B R A I N (49) (50) (50)A S T R O C Y T O d A 1 (2%)
SPECIAL S E N S E O R G A N S
* Z Y M B A L ' S G L A N D (49) (50) (50)S Q U A H O U S c -ELL C A R C I N O M A 2 (4%)
r t U S C U L O S K E L E T A L S Y S T E M
* B O N E ( 4 9 ) (50) (50)OSTEOSARCUMA 1 (2%)
* S K E L E T A L M U j C L E ( 4 9 ) (50) (50)O S T E O S A R C O f l A , I N V A S I V E 1 ( 2 % )
B O D Y C A V I T I E S
T U N I C A V A G I i i A L I S (49) (50) (50)
* NUMBER OF AuIJULS WITH TISSUE EXAMINED MICROSCOPICALLY* NUMBER OF AdlHALS NECROPSIED
43
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)
A I L 01KER S Y S T E M S
* M U L T I P L E O E o A N SM E S O T H E L I U M A , N05J lESOTHELl j r tA , M A L I G N A N T
A N I M A L DISPOSITION S U M M A R Y
A N I M A L S I N I T I A L L Y I N S T U D YN A T U R A L Di iATHdM O R I B U N D S A C R I F I C E
TOTAL ANIMALS KITH PRIMARY TUMORS*TOTAL PRIdARY TUMORS
TOTAL ANIMALS WITH BENIGN TUMORSTOTAL BENIGN TUMORS'
TOTAL ANIMALS WITH MALIGNANT TUMORSTOTAL MALIGNANT TUMORS
4790
4659
2428
50106
4780
2326
49100
4777
1822
TOTAL A N I M A L S W I T H S E C O N D A R Y T U M O R S * 3 1 1
T O T A L S E C O N D A R Y T U M O R S 3 2 1
TOTAL A N I M A L S WITH T U M O R S U N C E R T A I N -B E N I G N OR M A L I G N A N T 3 1
TOTAL U N C E R T A I N T U M O R S 3 1
TOTAL A N I M A L S W I T H T U M O R S U N C E R T A I N -- P B I M A R Y OR i-iETASTATIC
TOTAL UNCdHTAIN TUMORS
* P R I M A R Y TUHOBS: ALL TUMOflS EXCEPT S E C O N D A R Y T U M O R S* S E C O N D A R Y T U M O R S : METASTATIC T U H O R S OH T U M O R S I N V A S I V E INTO AN ADJACENT O R G A N
44
TABLE A2.
SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS
ADMINISTERED TITANIUM DIOXIDE IN THE DIET
ANIMALSANIMALSANIMALS
INITIALLY INNSCHOPSIED
STUDY
EXAMINED HISTOPATHOLOGICALLY
MATCHEDCONTROL
505050
LOW DOSE
505050
HIGH DOSE
504949
INTtGUMENTAKY SYSTEM
*SKINSQUAMOUS CELL CARCINOMA
*5UBCUT TISSUESQUAHOUSFIBROMA
CELL CARCINOMA
(50)1
(50)11
12%)
(2%)(2%)
(50)
(50)11
(2%)(2%)
(49)3
(19)
(6X)
RESPIRATORY SYSTEM
*LUNGS Q U A M O U S C E L L C A R C I N O M A , M E T A S T AA L V E O L A R / u f i O N C H I O L A R A D E N O M AALVEOLAR/ i iRONCHIOLAR C A R C I N O M A
(50)
2 (4X)1 (2%)
(50)
1 (2X)
(49)1 (2X)1 (2%)
HEMATOPOIETIC SYSTEM
*HULTIPLS O R G A N S (50)M A L I G . L Y M P H O M A , HISTIOCfTIC TYPEG R A N U L O C Y T I C L E U K E M I AM O N O C Y T I C L E U K E M I A 10 (2095)
tCJSEVICAL LYdPH NODE (50)S Q U A M O U S CELL C A R C I N O M A , METASTA
(50)2 (48)1 (2X)
10 (20X)
(50)
(49)
111 (2255)
(49)2 <4S)
CIRCULATORY SYSTEM
NONE
DIGESTIVE SYSTEM
tLIVER_ NEOPLASTIC NODULE,
(50) (49)
I NUMBER OF AiilMALS HITH TISSUE EXAMINED MICROSCOPICALLY* NUMBED OF ANIMALS NECROPSIED
45
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED)
MATCHEDCONTROL LOW DOSE HIGH DOSE
ffSTCMACHS Q U A M O U S CELL P A P I L L O M A
(50) (50) (48)1 (2%)
U R I N A R Y SYSTSd
N O N E
E N D O C R I N E SYSlEM
•PITUITARYC A R C I N O M A , NOSCHPOMOPHOjE A D E N O M AC H R O M O P H O o E C A R C I N O M A
• A D B E N A LCORTICAL *DENOMAF H E O C H R O M O C Y T O M A
I F T H Y R O I DFOLLICULAH-CELL A D E N O M AC-CELL A D E N O M AC-CSLL C A K C I N O M A
* P A N C R E A T I C ISLETSISLET-CELi , A D E N O M A
8EPRODUCTIVE SYSTEM
* M A M M A R Y G L A t M DA D E N O M A , u f U SA D B N O C A R C i N O B A , N O SC Y S T A D E N O i l A , N O SF I B R O A D E M O M A
*PREPUTIAL G L A N DC A R C I N O K A . N O SA D E N O M A , f c i O S
t U T E R U SC A R C I N O M A , NOSF I d R O M A
(U8)
28 (58%)
(50)
(48)2 (4%)
1 (2%)
(50)
(50)
1 (2«)1 (2%)
20 (UOSS)
(50)2 (4%)
(50)
(1*7)3 (6%)
26 (55%)
(49)2 (4%)1 ( 2%)
(47)
(50)1 ( 2 % )
(50)
2 (4&)
14 (28%)
(50)2 ( 4 % )1 (255)
(50)
1 12%)
(47)3 (6X)
31 (66%)1 (2%)
(19)
1 ( 2 X )
( 4 4 )
2 (5X)4 (9%)
(490
(49)1 (27.)2 (4%)
19 (39%)
(49)3 (b%)
(49)1 (251)
* N U M B E R OF A N I M A L S WITH TISSUE E X A M I N E D MICROSCOPICALLY* N U K B i i K O F A N I M A L S N E C R O P S I E D
*3fiAIN (48) (48) ( 4 9 )C A R C I N O M A , N G S , M B T A S T A T I C 2 (48) 2 (47.)S Q U A M O U S C E L L C A R C I N O M A . M E T A S T A 1 (2%)CHROHOPH00E C A R C I N O M A , MET AST ATI 1 (2%)G L I O M A , N O S 1 (2%)ASTROCYTOhA 1 (2X)
*SKELETAL MUSCLE (50) (50) (49)S A R C O M A , i iOS 1 (2%)
E C D Y C A V I T I E S
N O N E
A L L O T H E R S Y S i ' E M S
* NUMBER OF AuIMALS WITH TISSUE EXAMINED MICROSCOPICALLY* NUMBER OF AwIKALS N2C80PSIED
47
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED)
MATCHEDCONTROL LOW DOSE HIGH DOSE
SNIMAL DISPOSITION SUMMARY
ANIMALS INITIALLY IN STUDYNA T U R A L DEATHSMORIBUND SACRIFICESCHEDULED SACRIFICEACCIDENTALLY KILLEDTERMINAL SACRIFICEA N I M A L MIoSING
dS INCLUDES AUiOLYZED ANIMALS
50113
36
50122
36
50142
34
TUMOR S U M M A R Y
TOTAL ANIMALS KITH PRIMARY TUMORS*TOTAL PRIMARY TUMORS
TOTAL ANIMALS WITH BENIGN TUMORSTOTAL BENiGN TUMORS
TOTAL ANIMALS WITH MALIGNANT TUMORSTOTAL MALIGNANT TUMORS
4183
38
62
19
20
4386
3763
2023
4696
4166
24
30
TOTAL ANIMALS WITH SECONDARY TUMORS* 1 3 5TOTAL SECONDARY TUMORS 1 3 6
TOTAL ANIMALS WITH TUMORS UNCERTAIN-BENIGN OH MdLIGNANT 1
TOTAL UNCERTAIN TUMORS 1
TOTAL ANIMALS WITH TUMORS UNCERTAIN-PRIMARY OR tIETASTATICTOTAL UNCERTAIN TUMORS
* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS* SECONDARY TUMORS: METASTATIC TUMOKS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
48
APPENDIX B
SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
MICE ADMINISTERED TITANIUM DIOXIDE IN THE DIET
49
50
TABLE B1.SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE
ADMINISTERED TITANIUM DIOXIDE IN THE DIET
A N I M A L SA N I M A L SA N I M A L S
I N I T I A L L Y I N S T U D YN E C R O f S I E DE X A M I N E D HI STOP A T H O L O G I C A L L Y
MATCHEDCONTROL
504747
LOW DOSE
504949
.« — — — — — ™--"- -»
HIGH DOSE
504949
INTEGUMENTARY SYSTEM
* S K I NF I B R O M A
* S U B C U T T I S S U ES E B A C E O U S A D E N O M AF I B R O M AF I l i f i O S A R C X l M AH E M A N G I O S a R C G M A
.(«7)
(47)
4 (955)8 (1735)1 (2%)
(49)1 (2«)
(49)
3 ( 6 X )8 (16S)
(49)
( 4 9 )1 (2%)1 (2%)4 (835)
RESPIRATORY SYSTEM
# L U N GH E P A T O C E L j u U L A R C A R C I N O M A , M E T A S TA L V E O L A K / o R O N C H I O L A R A D E N O M AA L V E O L A R / j R O N C H I O L A R C A R C I N O M A
(46)
5 (1U)1 (2%)
(49)2 ( 4 % )2 (435)1 (235)
(49)1 (235)5 ( 1 0 X )
HEMAIOPOI2TIC SYSTEM
" K U L T I P LM A L I GflALIG
E O R G A N S. L Y f l f H O M A. L Y K r > H O B A
G R A N U L O C Y i ' I C L
» K E 3 F N T EH E M A NH EM A NH A L I G
,
r
L Y M P H O C Y T I CH I S T I O C Y T I C
T Y P ET Y P E
(47)41
(935)(2%)
E U K E M I A
H I C i.. N O D EG I O M i iGios«ificoa. L Y H r H O M Ai H I S T I O C Y T I C T Y P E
(47)
1 ( 2 X )
(49)232
(48)21
( 4 X )(635)(4%)
(435)(2X)
( 4 9 )
5
( 4 8 )
(103)
C I R C U L A T O R Y S Y S T E M
(4b)HEMA| i2 I2 S
(49) ( 4 9 )
a NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY* NUMBER OF AnIMALS NECROPSIED
51
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED)
MATCHEDCONTROL LOW DOSE HIGH DOSE
D I G E S T I V E S Y S T E M
* I N T E S T I N A L T R A C T (47) (49) ( 4 9 )C A R C I N O M A , N O S 1 (2%)
TOTAL ANIMALS WITH SECONDABY TUMOBSfTOTAL SEOJNDA8Y TUMORS
(47)1 (2%)
50
17
132
2936
1010
2225
(49)
5010
40
2537
810
2227
22
(49)
5010
40
2838
1112
2326
11
TOTAL ANIHA.LS IITH TUBOES U N C E R T A I N -B E N I G N OR M A L I G N A N T 1
TOTAL U N C E R T A I N TUMORS 1
TOTAL A N I B A L S WITH T U M O R S U N C E R T A I N -Pf i lMARY OH SETASTATIC
TOTAL U N C E R T A I N TUMORS
* P R I M A R Y TUMORS: ALL TUMOBS EXCEPT SECONDARY TUBORSt S E C O N D A B Y T U M O R S : BET&STATIC TOBOHS OB TUMOBS I N V A S I V E IMTO AN ADJACENT ORGAS
53
TABLE B2.
SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICEADMINISTERED TITANIUM DIOXIDE IN THE DIET
A N I M A L SA N I M A L Si N I H A L SJ N I M A L S
I N I T I A L L Y I N S I U D YH I S S I u GNEC80i?SI2DE X A M I a i D H I S T O P A T H O L O G I C A L L Y
MATCHEDCONTROL
501
4949
LOW DOSE
50
5050
HIGH DOSE
50
5050
INTEGUMENTARY SYSTEM
*SUBCUT TISSUETRICHOEPIiHELIOMAFIBROSARCdMA
(49 ) (50)
2 ( 4%)
(50)1 (2%)
5 E S P I E A T O R Y S Y S T E M
# L U N GA D E N G C A R C i N O H A , NOS, M E I A S T A T I CA L V E O L A R / r i R O N C H I O L A R A D E N O M AA L V E O L A R / j R O N C H I O L A B C A R C I N O M AF I B R O S A R C d M A , M E T A S T A T I CL E I O M Y O S A t t C O M A , M E T A S T A T I C
( 4 9 )
1 (2%)
1 (2%)
(50)
1 ( 2 X )1 < 2 X )1 (2%)
(50)1 ( 2 % )3 (6X)1 (2%)
H E M A T O P O I E T I C SYSTEM
*MULTIPLE O R G A N S (49 )M A L I G . L Y M P H O M A , L Y M P H O C Y T I C T Y P E 6 (12»)M A L I G . L Y M P H O M A , HISTIOCYTIC T Y P E 12 (24%)M A L I G N A N T L Y M P H O M A , M I X B D T Y P E 2 (4%)G R A N U L O C Y I I C L E U K E M I A
# S P L E E N (49)H E M A N G I O S A B C O M AH A L I G . L Y M P H O B A , HISTIOCYTIC T Y P E
•CERVICAL LYuPH NODE (48)HEMANGIOSAECOMA 1 (2%)
*THYMUS (23)FIBROSARCUHAj_META STATIC
(50)4 (8»)7 (143t)
(50)1 (28)
(47)
(27)
(50)7 (1455)4 (8X)
2 (4%)
(50)
1 (2%)
(47)
(34)
« NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY* NUMBER OF ANIMALS NECROPSIED
54
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED)
C I R C U L A T O R Y Si 'STtfl
t H E A H TH E M A N G I O M f t
C i G E S T I V E S Y S I E K
S L I V E RH E P A T O C E i ^ U L A S C A R C I N O M A
• P A N C R E A SF I B R O S A R C O M A , H E T A S T A T I C
*STOBACHL E I O B Y O S A a C O M A
* L A R G E I N T E S T I N EL E I O M Y O S A H C O M A , B E T A S T A T I C
U B I N A E Y SYSTEd
# K I D N E YT U B U L A R - C i i L L A D E N O C A R C I NOB AL E I O M Y O S A a C O B A , M E T A S T A T I C
& U R I N A R Y B L A D D E RL E I O M Y O S A R C O M A , M E T A S T A T I C
ENDOCRINE SYSi'EB
# P I T O I T A R YC H B O M O P H O o E A D E N O M A
# T H Y R O I DFOLLICULAH-CELL A D E N O M AC-CELL A D E N O M A
R E P R O D U C T I V E S Y S T E M
* M A M B A R Y G L A N DA D J E N O C A H C I N O M A . NOS
MATCHEDCONTROL
( 4 9 )
(49)1 (255)
(49 )
(48)1 (2X)
(48)1 (2%)
(49)
1 (2X)
(47)1 (2%)
(33)3 (9»)
(43)3 (7S)
(49)_ 1 12$L
LOW DOSE
(50)
(50)3 (6%)
(50)1 (255)
(50)
(50)
(50)1 (235)
(45)
(tO)4 (10«)
(41)
(50)1.128) ,
HIGH DOSE
( 5 0 )1 ( 2 % )
(50)3 ( 6 % )
(50)
(49 )
( 4 9 )
(50)
(45)
(33)2 (6%)
(44)
1 (28)
(50)3 16%}
t NUMBER OF ANIBALS WITH TISSUE EXAMINED MICROSCOPICALLY* NUBBER OF AtUBALS NECROPSIED