Henry J. Trochimowicz

Inhalation toxicity study on rats exposed to titanium tetrachloride atmospheric hydrolysis products for two years

Rats were exposed to TiCl4 hydrolysis products by inhalation exposure at aerosol concentrations of 0, 0.1, 1.0, and 10 mg/m3 for 6 hr/day, 5 days/week for 2 years. There were no abnormal clinical signs, body weight changes, or excess mortality in any exposed groups. No pathological changes other than a mild rhinitis were observed at 0.1 mg/m3. At 1.0 mg/m3, the incidence of mild rhinitis and tracheitis was increased. The lungs showed a minute dust-laden macrophage (dust cell) reaction with slight Type II pneumocyte hyperplasia in alveoli adjacent to the alveolar ducts. The pulmonary response at the 1.0 mg/m3 satisfied the biological criteria for a nuisance dust. At 10 mg/m3, extrapulmonary particle deposition occurred in the tracheobronchial lymph nodes, liver, and spleen without any tissue response. An increased incidence of rhinitis, tracheitis, and dust cell response with Type II pneumocyte hyperplasia, alveolar bronchiolarization, foamy dust cell accumulation, alveolar proteinosis, cholesterol granuloma, and focal pleurisy was also observed. The pulmonary lesions developed in the alveolar duct region where dust cells had accumulated and had provoked a chronic tissue response. In addition, a few well-differentiated, cystic keratinizing squamous carcinomas were developed from alveoli showing bronchiolarization with squamous metaplasia in the alveolar duct region. No tumor metastasis was found in other organs. The lung tumors were a unique type of experimentally induced tumor and have not been seen usually in man or animals. Therefore, the relevance to man of this type of lung tumor is highly questionable.

Transmigration of titanium dioxide (TiO2) particles in rats after inhalation exposure.

Titanium dioxide (TiO2) has been used extensively in the manufacturing of white pigment and has generally been regarded as a nuisance dust in animals and man. After inhalation exposure, little is known about transmigration routes and potential toxic effects of translocated particles in other organs. In order to answer these questions, rats were exposed to TiO2 by inhalation exposure at concentrations of 0, 10, 50, and 250 mg/m3 for 2 years. A few free particles were retained in the nasal and tracheobronchial epithelium without any cellular damage, but aggregates of dust-laden macrophages (dust cells) were found in the lymphoid tissue of the submucosa. Inhaled particles were mostly engulfed by alveolar macrophages and confined sharply to the alveolar duct region at 10 and 50 mg/m3, while dust cells were scattered throughout alveoli at 250 mg/m3. A fraction of the inhaled particles was retained in the membranous pneumocytes and interstitial macrophages. A dense accumulation of dust cells was found in the perivascular and peribronchial lymphoid tissue. Some dust cells entered peribronchial lymphatics or pulmonary blood vessels and the general circulation. Dust cells in the hyperplastic peribronchial lymphoid tissue were exposed directly in the luminal surface of the airways and were subsequently eliminated via airways. Massive dust deposition was observed in the tracheobronchial lymph nodes. Dust transmigration was markedly reduced in the cervical lymph nodes, and only a trace amount of dust particles was found in the mesenteric lymph nodes. Some dust cells entered either blood or lymphatic vessels in the lymph nodes and then migrated into the general circulation. The incidence of extrapulmonary dust deposition in the liver or spleen was increased in a dose-related fashion similar to the lung dust burden. Since there was no tissue response to translocated particles in the lymph nodes, spleen, or liver, potential adverse health effects appear to be negligible.

Acute and subchronic toxicity of 1,1-dichloro-1-fluoroethane (HCFC-141b).

The acute and subchronic toxicity of 1,1-dichloro-1-fluoroethane (HCFC-141b), a CFC alternative, was evaluated in several acute and subchronic studies to assist in establishing proper handling guides. Data from acute toxicity studies in rats and rabbits demonstrated that HCFC-141b has very low acute toxicity. HCFC-141b was not a skin irritant, but was a mild eye irritant, in rabbits and was not a skin sensitizer in guinea pigs. Skin application of HCFC-141b to rabbits at 2000 mg/kg body weight produced no adverse effects. Oral administration at 5000 mg/kg body weight did not cause any deaths or clinical signs of toxicity in rats. The 4-hr LC50 for HCFC-141b was about 62,000 ppm in rats. Repeated exposures of rats for 6 hr/day, 5 days/wk for up to 90 days at concentrations of 2000, 8000 or 20,000 ppm did not result in significant adverse effects. Minor, but dose-dependent, reductions in body weight were observed in male and female rats during the 90-day study. Decreased responsiveness was also observed in rats but only at 20,000 ppm. An increase in serum cholesterol or triglycerides was observed in male and female rats at 20,000 ppm, and in males at 8000 ppm. No specific organ pathology was noted in these subchronic inhalation studies. The no-observable-adverse-effect level (NOAEL) from these studies was 8000 ppm. Results from other studies demonstrate that HCFC-141b was not neurotoxic in rats. As with trichlorofluoroethane (CFC-11), a cardiac sensitization response to an intravenous epinephrine challenge occurred in dogs with HCFC-141b at 5000 ppm and higher concentrations in experimental screening studies.

Effects of Inhaled Chromium Dioxide Dust on Rats Exposed for Two Years

Rats were exposed by inhalation to chromium dioxide (CrO2) dust at design concentrations of 0, 0.5 mg/m3 (stabilized and unstabilized, respectively), or 25 mg/m3 (stabilized) for 6 hr/day, 5 days/week for 2 years. No dust-exposure-related pathological changes other than lung lesions were observed in all exposed rats. There were no significant differences in pulmonary responses between unstabilized and stabilized CrO2 at the 0.5 mg/m3 exposure level. The lungs showed minute dust deposition in the alveoli adjacent to the alveolar ducts, but maintained an intact general architecture. The pulmonary responses satisfied the biological criteria for a nuisance dust. At 25 mg/m3, dust deposition was sharply confined to the alveoli in the alveolar duct region. Alveolar walls enclosing dust-laden macrophage (dust cell) aggregates were thickened with hyperplastic Type II pneumocytes and slightly collagenized fibrosis. Alveoli adjacent to the terminal bronchioles were lined with bronchiolar epithelium (alveolar bronchiolarization). In addition, lungs showed foamy macrophage response, cholesterol granulomas, alveolar proteinosis, and minute fibrotic pleurisy. These pulmonary lesions occurred predominantly in female rats. Of 108 female rats, 6 developed keratin cysts and 2 had cystic keratinizing squamous cell carcinoma (CKSCC). None of 106 male rats had either a keratin cyst or a CKSCC. The lung tumors developed from metaplastic squamous cells in the areas of alveolar bronchiolarization at the alveolar duct region. The lung tumors were well differentiated and devoid of characteristics of true malignancy. The CKSCC is an experimentally induced, unique tumor type and is different from the type of spontaneous lung tumor seen in man or animals. The relevance to man of this type of lung tumor appears to be negligible.

Blood Levels of Fluorocarbon Related to Cardiac Sensitization

Unanesthetized beagle dogs were exposed to concentrations of fluorocarbon-11 and fluorocarbon-12 which had been reported to produce cardiac sensitization. During and after the exposure, arterial and venous blood samples were obtained for fluorocarbon analysis. The blood concentration rose rapidly during the first few minutes of the 10-minute exposure and more slowly thereafter. When the exposure was terminated, there was a rapid initial fall in the blood concentration followed by a more prolonged decline. A definite arterial-venous difference was found. Despite a tenfold difference in the inspired concentration of the two compounds, the blood concentrations associated with cardiac sensitization were similar for both agents. The average blood concentration (µg/ ml) associated with exposure to levels known to sensitize the beagle heart was: 28.6 arterial and 19.7 venous for fluorocarbon-11; and 35.3 arterial and 22.8 venous for flurocarbon-12.

Acute, Subchronic, and Developmental Toxicity and Genotoxicity of 1,1,1-Trifluoroethane (HFC-143a)

The toxicity potential of 1,1,1-trifluoroethane (HFC-143a), a CFC alternative, was evaluated in several acute, subchronic, and developmental toxicity studies by the inhalation route and in genotoxicity studies. HFC-143a has a very low acute inhalation toxicity potential as shown by a 4-hr LC50 of > 540,000 ppm in rats. HFC-143A has a low potential to induce cardiac sensitization in experimental screening studies in dogs; only the highest concentration tested--300,000 ppm--elicited a cardiac sensitization response. In an initial 4-week nose-only inhalation study, male and female rats were exposed 6 hr/day, 5 days/week at concentrations of 0, 2000, 10,000, or 40,000 ppm. Females showed no evidence of toxicity at any exposure level; male rats did exhibit degenerative changes only in the tests at all exposure levels. However, because of exposure system irregularities, which resulted in excessive temperature conditions and stress in the HFC-143a-exposed groups, the study was repeated in male rats exposed by whole-body inhalation. In this repeat study no toxicity was observed at < or = 40,000 ppm. Moreover, a subsequent 90-day whole-body inhalation study in rats exposed 6 hr/day, 5 days/week at 0, 2000, 10,000, or 40,000 ppm resulted in no evidence of toxicity at any exposure concentration. The results of the second 4-week and the 90-day studies using whole-body exposures indicate that the findings from the first 4-week study were related to the stress induced by excessive temperatures and nose-only restraint. Therefore, the no-observed-effect level (NOEL) for rats repeatedly exposed up to 90 days was considered to be 40,000 ppm. In developmental toxicity studies with rats and rabbits, an increase in visceral variations or skeletal malformations was observed, respectively, at HFC-143a concentrations of 2000, 10,000 or 40,000 ppm (rat) or at the low and high concentrations (rabbit). Because of the unusually low control incidence of variations (1.6% per litter in the control versus 6.8-16.8% for historical control values), the lack of a clear dose-response relationship, and the lack of other developmental effects, these findings were not considered related to HFC-143a exposure. In addition, results from genotoxicity studies (Ames, chromosomal aberration with human lymphocytes, mouse micronucleus) demonstrated that HFC-143a was not mutagenic.

13-week drinking water toxicity study of hydrogen peroxide with 6-week recovery period in catalase-deficient mice.

A GLP OECD guideline study was conducted to evaluate the subchronic toxicity of hydrogen peroxide (HP) when administered continuously in the drinking water of catalase-deficient (C57BL/6N) mice and reversibility of toxic effects. Groups of mice (15/sex/group) received solutions of 0, 100, 300, 1000 or 3000 ppm HP in distilled water for 13 weeks; five/sex/group continued on untreated distilled water for an additional 6 weeks. Animals drinking 3000 ppm HP exhibited depressed water and food consumption and body weight. Females drinking 1000 ppm HP had reduced water consumption with intermittent effects on food consumption, but no body weight effects. HP administration did not produce any mortality, clinical signs, hematological effects or organ weight effects on brain, liver, kidneys, adrenals, testes, heart or spleen. Total protein and globulin were depressed among high dose males. Mild to minimal duodenal mucosal hyperplasia was noted in animals receiving 1000 and 3000 ppm HP and one male receiving 300 ppm for 13 weeks. There were no other histopathological findings. All effects noted during the treatment period, including the duodenal hyperplasia, were reversible during the 6-week recovery period. Females dosed with 300-3000 ppm HP during the treatment period showed increased water consumption during the recovery period. The no-observed-effect level (NOEL), based on duodenal mucosal hyperplasia, is 100 ppm in drinking water or 26 and 37 mg/kg/day HP, respectively, for males and females.

Toxicological evaluation of 1,1,1,2,2-pentafluoroethane (HFC-125)

Acute, subacute, and subchronic inhalation toxicity studies, developmental toxicity studies, a cardiac sensitization evaluation, and mutagenicity assays were conducted with pentafluoroethane (HFC-125). In the acute study, rats were exposed to a single concentration of 800,000 ppm for 4 hr. Ataxic gait and abnormal respiration were observed during exposure but not after exposure. There was no mortality or other signs of toxicity. Repeated exposures of rats to 50,000 ppm, 6 hr/day, 5 days/week for either 4 or 13 weeks elicited no effects on body weight, food consumption, clinical signs, hematology, biochemistry, urinalysis, organ weight, or tissue morphology. Positive evidence of cardiac sensitization in response to an intravenous epinephrine challenge in dogs was seen at 100,000 ppm and above, but not at 75,000 ppm. HFC-125 was not mutagenic in Salmonella typhimurium and Escherichia coli strains at concentrations of 20 to 100% (v/v) with and without activation. No evidence of clastogenic activity was observed in cultured Chinese hamster ovary (CHO) cells or human lymphocytes at < or = 70% HFC-125 when treatments were conducted for 3-4 hr with activation or for 24 and 48 hr (human lymphocytes only) without activation. However, a statistically significant increase in chromosomally aberrant cells was observed in CHO cells at 60% HFC-125 when treatment without activation was extended to 48 hr. The biological significance of this effect is questionable since signs of severe toxicity were also present. In vivo, no micronuclei were induced in mouse bone marrow at concentrations as high as 600,000 ppm HFC-125 for a 6-hr exposure. In addition, HFC-125 did not induce embryotoxic or teratogenic effects in either the rat or the rabbit at exposure concentrations as high as 50,000 ppm.

Cardiac sensitization: methodology and interpretation in risk assessment.

An increased sensitivity of the heart to endogenous epinephrine (adrenaline), a phenomenon referred to as cardiac sensitization, has long been recognized as a risk during exposure to hydrocarbons, principally halogenated hydrocarbons. Cardiac sensitization, which can result in serious arrhythmia and death, requires a certain critical blood level of both the sensitizing agent and epinephrine. The original research and methods utilized an exogenous epinephrine challenge during inhalation exposure to a chemical to assess cardiac sensitization potential in Beagle dogs. These screening tests were developed about 30 years ago, although in the last 15 years some modifications of these methods have occurred in response to testing chlorofluorocarbon (CFC) replacements. Results from these experimental cardiac sensitization studies have been used for semi-quantitative risk evaluation for occupational exposures but now are being used more quantitatively for regulatory purposes. The risks associated with cardiac sensitization from CFC replacements are unknown but expected to be low based on cardiac sensitization studies in the 1970s where dogs were made to generate their own adrenaline. With the advent of physiologically based pharmacokinetic (PBPK) modeling, greater emphasis is being placed on quantitative risk assessment for cardiac sensitization. In this investigation, we have examined the various methodologies used for detection of cardiac sensitization and discussed their limitations and advantages. In addition, we examined the potential concerns involved in using experimental cardiac sensitization data and PBPK modeling to predict exposure scenarios.

Pulmonary response to inhaled hexamethylphosphoramide in rats

Abstract Dose-related nasal tumors, rhinitis, nasal epithelial degeneration, squamous metaplasia, and dysplasia were found in rats exposed to hexamethylphosphoramide (HMPA) by inhalation at 50, 100, 400, and 4000 ppb ( v v ) for 6 to 24 months. No pathological lesions were observed in rats exposed for 24 months at 10 ppb. One adenocarcinoma and one papilloma were found in the tracheal epithelium at 400 and 4000 ppm, respectively. Tracheitis, degeneration of the tracheobronchial epithelium, and murine pneumonia were dose related at the levels of 100, 400, and 4000 ppb but not at the levels of 10 and 50 ppb. Keratinized squamous metaplasia developed in the trachea at 4000 ppb. Damage to the tracheobronchial epithelium gradually decreased toward peripheral air passages and no pathological changes were observed in the alveolar walls. Microscopically, the tracheobronchial epithelium was lined with degenerative cells and regenerative cuboidal or flattened cells resembling squamous cells. Under electron microscopy, ciliated epithelial cells appeared to be sensitive to HMPA and showed degenerative changes with abnormal cilia. The desquamated tracheobronchial epithelium was initially repaired by undifferentiated glandular and microvillous cells, but not by ciliated cells. Extensive destruction of the mucociliary apparatus in the air passages appeared to be responsible for the increased incidence of the murine pneumonia in rats exposed to HMPA.