DMSO toxicity

Chuck Miller rellim at MAILHOST.TCS.TULANE.EDU
Thu Oct 15 15:52:53 EST 1998

Here are a few references. DMSO is remarkable stuff--- I have always been
amazed that cells survive well in 10% DMSO.

Toxicology 1988 Nov 14;52(1-2):165-75

Prevention of acetaminophen-induced hepatotoxicity by dimethyl sulfoxide.

Park Y, Smith RD, Combs AB, Kehrer JP

Division of Pharmacology and Toxicology, College of Pharmacy, University of
Texas, Austin 78712-1074.

Dimethyl sulfoxide (DMSO) has previously been shown to protect against
acetaminophen (APAP)-induced hepatotoxicity, but the mechanism of this
effect was not clear. Treatment of mice with 1 mg/kg DMSO 4 h before 250
mg/kg APAP resulted in significantly less hepatotoxicity than with APAP
alone, as measured by serum glutamic pyruvic transaminase (SGPT) content 24
h after APAP. Protection was also evident when 1 ml/kg DMSO was given 4,
but not 8 h after 250 mg/kg APAP. The APAP-induced depletion of liver
glutathione was prevented in mice pretreated with DMSO, although DMSO alone
had no effect on liver glutathione levels. The hepatic concentration of
cytochrome P-450 (P450) 4 h after treatment of mice with 1 ml/kg DMSO, was
significantly decreased compared to saline-treated animals. However, while
this DMSO pretreatment significantly decreased the activity of cytochrome
P-450-linked aminopyrine-N-demethylase, it increased the activity of
aniline hydroxylase. Covalent binding of [14C]APAP to hepatic protein in
vivo was significantly decreased in mice pretreated with DMSO. Covalent
binding of [14C]APAP to hepatic microsomal protein in vitro was not
significantly altered after in vivo treatment with DMSO. However, the
presence of DMSO in the in vitro incubation mixture significantly decreased
covalent binding of [14C]APAP in a dose-dependent manner compared to
microsomal fractions from untreated, saline-treated or DMSO pretreated
animals. These data suggest that the DMSO-induced alterations in cytochrome
P-450 content and activity may not be the cause of the observed protective
action of this chemical. The ability to competitively inhibit APAP
bioactivation or to directly scavenge free radicals produced during APAP
metabolism, including the activated species which covalently binds to
protein, may account for the hepatoprotection afforded by DMSO.

Drug Metabol Drug Interact 1988;6(3-4):413-24

Mechanism of inhibition of hepatic bioactivation of paracetamol by
dimethyl sulfoxide.

Jeffery EH, Arndt K, Haschek WM

Institute for Environmental Studies, University of Illinois, Urbana 61801.

Prior work has shown that DMSO inhibits paracetamol hepatotoxicity. In this
paper we show that DMSO and its reduced metabolite dimethyl sulfide (DMS)
can inhibit in vitro hepatic  dimethylnitrosamine N-demethylase. We also
show that DMSO can inhibit in vivo production of glutathione conjugates of
paracetamol. Glutathione is known to conjugate the bioactivated form of
paracetamol. Also, the isozyme of cytochrome P-450 responsible for
dimethylnitrosomine N-demethylase, cytochrome P-450j, is thought
responsible for paracetamol bioactivation. We therefore propose that DMSO
inhibits paracetamol hepatotoxicity due to inhibition of cytochrome
P-450j-dependent paracetamol bioactivation by DMSO and its metabolite DMS.

Arzneimittelforschung 1994 Apr;44(4):566-70

Acute intravenous toxicity of dimethyl sulfoxide, polyethylene
glycol 400, dimethylformamide, absolute ethanol, and benzyl
alcohol in inbred mouse strains.

Montaguti P, Melloni E, Cavalletti E

Research Centre, Boehringer Mannheim Italia, Monza, Milan, Italy.

Acute intravenous toxicity of some solvents, i.e. dimethyl sulfoxide
(DMSO), polyethylene glycol 400 (PEG 400), dimethylformamide (DMF),
absolute ethanol (EtOH) and benzyl alcohol (BeOH), was determined in three
inbred (CD2F1, B6D2F1 and C57BL/6N) mouse strains used in many preclinical
tests, mainly in oncology and toxicology. Haemolytic and precipitation
potential tests in vitro were performed to assess the blood compatibility
of the investigated solvents and its relationship with the observed
symptoms. The single tested solvents did not show any major differences in
acute toxicity in the three tested strains with the exclusion of DMSO (less
toxic in CD2F1) and BeOH and EtOH (less toxic in B6D2F1). The tested dose
ranges in the three strains (in ml/kg) were 1.0-5.66 for DMSO, 2.0-8.0 for
PEG 400, 1.0-4.0 for DMF, 0.75-4.24 for EtOH, 0.025-0.4 for BeOH. The
lowest tested dose was a safe dose and the highest one was the dose causing
mortality in no more than half the animals in each group. The in vitro
results suggest avoiding the use of BeOH (which also is more toxic than the
other solvents in the in vivo test) and DMSO and using PEG400, EtOH and DMF
even though the latter induced a body weight decrease in the B6D2F1 mouse
strain. As a general conclusion, dilution of these solvents in water is
suggested to ameliorate their blood compatibility and the use of doses not
higher than the lowest dose tested in this study is recommended.

Toxicol Appl Pharmacol 1996 Oct;140(2):296-314

An analysis of dimethylsulfoxide-induced action potential block: a
comparative study of DMSO and other aliphatic water soluble solutes.

Larsen J, Gasser K, Hahin R

Northern Illinois University, Biological Sciences Department, DeKalb 60115,

A series of water soluble aliphatic solutes were chosen for study. Fifty
percent effective doses (ED50) to block propagated compound action
potentials (AP's) were obtained by examining dose-response relations for
each solute. All solutes used were liquids at room temperature and are
typically used as solvents. The solutes studied were dimethylsulfoxide
(DMSO), dimethylformamide (DMF), dimethylacetamide, acetone, and
hexamethylphosphoramide (HMPA); the octanol/water partition coefficients
for these test substances form an ordered sequence that increased 40-fold
from DMSO to HMPA. AP's were recorded from desheathed frog sciatic nerves
using the sucrose-gap technique; test solutes were added to Ringer's
solution and applied externally to the nerve. ED50's for the solutes could
be predicted as a function of the molar volume (dV/dn), polarity (P), and
the hydrogen bond acceptor basicity (beta). Voltage-clamp experiments
employing the vaseline-gap technique on single muscle fibers showed that
each solute reduced Na+ current with little change in their kinetics at all
voltages studied. Experiments using DMSO or DMF showed that Na+ channel
block alone is insufficient to explain the respective ED50 values of AP
block. Experiments conducted using a chloride transport-sensitive membrane
fluidity assay, using rat pancreas secretory granules, suggested that each
of the solutes act to increase membrane fluidity at doses below and above
ED50 values. Light microscopic observations of fixed thick sections of
whole nerves previously exposed to DMSO or DMF show structural changes;
however, ED50 values cannot be simply explained by osmotic alterations of
nerve structure. ED50's are likely to be produced by a combination of
effects including osmotically induced nerve structural changes, ion channel
block, and fluidity changes. The toxicity (lethal doses or toxic
concentrations) of each of these five solutes correlates well with the ED50
and could be predicted as a function of dV/dn, P, and beta.

Cryobiology 1996 Jun;33(3):354-62

Cryopreservation of isolated rat islets of Langerhans in the presence
of ethylene glycol or dimethyl sulfoxide: evaluation of toxicity and
the dynamic pattern of subsequent insulin release in vitro.

Sakonju I, Taura Y, Inayoshi Y, Suzuki T, Takimoto K, Nakaichi M, Nakama S

Department of Veterinary Surgery, Yamaguchi University, Japan.

The toxic effect of ethylene glycol (EG) on the pattern of dynamic insulin
release from rat pancreatic islets with or without freezing was
investigated in comparison with that of dimethyl sulfoxide (Me2SO). Sixty
islets were perifused (1 ml/min) consecutively with D-glucose (1.67 mM for
30 min followed by 16.7 mM for 60 min and 1.67 mM for 60 min) after
exposure to 2.0 M EG or Me2SO for 1 h at either 22 or 0 degrees C. During
the second period of perifusion, the insulin output from islets exposed to
Me2SO or EG at 22 degrees C decreased to 53 and 51% of that from nontreated
control islets, respectively. On the other hand, the islets exposed to EG
at 0 degrees C exhibited 86% of the control insulin output under the same
perifusion conditions, and this appeared to be higher than that of islets
exposed to Me2SO (60%) at 0 degrees C. Frozen islets, after exposure to 2.0
M EG or Me2SO for 1 h at 0 degrees C, responded positively to 16.7 mM
D-glucose, and the typical biphasic pattern of insulin secretion was
observed. The insulin output from these islets during the second period of
perifusion was not comparable to that from unfrozen control islets. In
particular, the mean insulin output of EG-cryopreserved islets during the
second period accounted for 99% of that from unfrozen control islets. The
present findings suggest the possible use of EG as an alternative
cryoprotectant to Me2SO.

Free Radic Biol Med 1996;20(1):129-38

Peroxyl radical formation in aqueous solutions of
N,N-dimethylformamide, N-methylformamide, and dimethylsulfoxide
by ultrasound: implications for sonosensitized cell killing.

Misik V, Riesz P

Radiation Biology Branch, National Cancer Institute, National Institutes of
Health, Bethesda, MD, USA.

Sonodynamic therapy, which refers to a synergistic effect of drugs and
ultrasound, is a promising new modality for cancer treatment. The
sonodynamic effect was found for a number of structurally unrelated
compounds, and the underlying mechanisms are still unknown. Recently,
Jeffers et al. (J. Acoust. Soc. Am. 97:669-676; 1995) have shown that the
sonodynamic action of nontoxic concentrations of N,N-dimethylformamide
(DMF), N-methyl formamide (MMF), and dimethylsulfoxide (DMSO) combined with
ultrasound, on killing of cultured HL-60 human promyelocytic leukemia
cells, and attributed this toxic effect to unknown short lived reactive
species produced from these solutes by ultrasonic cavitation. Using the
spin trap 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS) in
nitrogen-saturated aqueous solutions of DMF, MMF, or DMSO exposed to 50 kHz
ultrasound, we detected formation of .CH3 and .CH2N(CH3)CHO radical adducts
for DMF, mostly .CH2NHCHO adducts for MMF, and .CH3 adducts for DMSO. These
radicals were formed either by reactions of the solutes with
ultrasound-generated .H and .OH radicals (such as .CH2R-type radicals in
DMF and MMF, and .CH3 radicals in DMSO), or by direct pyrolysis of the weak
bonds in the solute molecules (e.g., .CH3 radicals from DMF). In
air-saturated sonicated solutions these carbon centered radicals were
converted to the corresponding peroxyl radicals and spin trapped with
5,5-dimethyl-1-pyrroline-N-oxide (DMPO); .OOCH2N(CH3)CHO radicals were
identified in DMF, .OOCH2NHCHO radicals in MMF, and .OOCH3 radicals in DMSO
solutions. We suggest that these radical species by virtue of their longer
lifetimes and higher selectivity, compared to .OH radicals, which are also
formed in sonicated solutions, are the species responsible for sonodynamic
cell killing by the combined effect of ultrasound with DMF, MMF, or DMSO.

Dig Dis Sci 1995 Feb;40(2):419-26

Toxic effects of cholelitholytic solvents on gallbladder and liver. A
piglet model study.

Chen CY, Chang KK, Chow NH, Leow TC, Chou TC, Lin XZ

Department of Internal Medicine, National Cheng Kung University, Tainan,

We evaluated the toxic effects of four currently used chemolytic
solvents--dimethyl sulfoxide (DMSO, 99%), ethyl propionate (EP, 99%),
tetrasodium ethyl-dimethyl tetraacetate (4Na-EDTA, 2%, pH 11), and methyl
tert-butyl ether (MTBE, purity = 99.5%) in an animal model. Each solvent
was tested in nine farm piglets (Landrace), weighing between 20 and 25 kg.
A solvent-resistant catheter was inserted transhepatically into the
gallbladder (GB) using sonographic guidance 24 hr prior to each experiment.
Seventy-five milliliters of each solvent was infused over 3 hr into the
gallbladder. The following day, a laparotomy was performed in order to
assess for possible damage to the liver, GB, bile ducts (BD), or
intestines. The GB and liver were resected and their histology examined.
The following pathologic grades were assigned to GB, BD, and liver
specimens to describe the tissue damage: normal (0), mild (1), moderate
(2), and severe (3). We found that DMSO had the highest score on
gallbladder and bile duct injury (49, 3), followed by EP (36, 2), EDTA (14,
1) and MTBE (16, 0), respectively; the difference in gallbladder damage was
statistically significant. Very mild hepatocyte damage was present in the
DMSO (2) and MTBE (2) groups. The administration of EP and EDTA resulted in
no liver injury at all. Piglets within each treatment group suffered from
varying degrees of tissue injury. No deaths were attributed to the
administered solvents. We concluded that DMSO, EP, EDTA, and MTBE do not
have serious local toxic effect on the GB, BD, and intestine; nor do they
lead to severe hepatotoxicity.

In Vitro Cell Dev Biol 1991 Aug;27A(8):633-8

Toxicity in vital fluorescence microscopy: effect of
dimethylsulfoxide, rhodamine-123, and DiI-low density lipoprotein
on fibroblast growth in vitro.

Crawford JM, Braunwald NS

Department of Pathology, Brigham and Women's Hospital, Boston,
Massachusetts 02115.

Fluorescence microscopy performed on living cells is a valuable technique
for elucidating patterns of cell growth in vitro over artificial
biomaterials such as vascular grafts, and for in vivo studies such as
identification and treatment of atherosclerotic plaques. Two fluorescent
dyes of particular value for vital fluorescence studies are Rhodamine-123
and 3,3'-dioctadecylindocarbocyanine-labeled low density lipoprotein
(DiI-LDL). We examined the toxicity of these two dyes and of
dimethylsulfoxide (DMSO), a solvent used in Rhodamine-123 studies, on the
growth of MRC5 human fetal fibroblasts in monolayer culture. Two parameters
of cell growth were quantitated: Cell number (a measure of proliferation),
and cell area (a measure of cell spreading), based on microscopic images
obtained at the start and end of a 48-h growth period after brief exposure
(0.5 h) to test solutions. We found that the recommended solvent for
solubilization of Rhodamine-123, DMSO, caused cessation of cell
proliferation and actual reduction in the area covered by adherent
fibroblasts at concentrations of as low as 0.1% (vol:vol). Rhodamine-123
made up from an aqueous stock solution modestly retarded proliferation and
spreading, and there was no significant effect of DiI-LDL on these
parameters over prolonged periods of exposure (up to 24 h) in culture.
These results demonstrate that the most toxic substance for growing
fibroblasts was the solvent DMSO. We conclude that both the solvent vehicle
and fluorescent dye should be carefully examined for potential toxicity
before such dyes are used for vital fluorescence studies of living cells.

J Leukoc Biol 1995 Jan;57(1):141-51

Polar agents with differentiation inducing capacity potentiate tumor
necrosis factor-mediated cytotoxicity in human myeloid cell lines.

Depraetere S, Vanhaesebroeck B, Fiers W, Willems J, Joniau M

Interdisciplinary Research Center, Laboratory of Biochemistry, Katholieke
Universiteit, Kortrijk, Belgium.

Cotreatment or pretreatment of several human myeloid cell lines (KG1, HL60,
U937, THP1) with the differentiation inducer DMSO was found to potentiate
the antiproliferative and cytotoxic effects of TNF. In addition,
TNF-resistant monocytic cell lines could be sensitized to TNF cytotoxicity
by DMSO treatment. Other highly polar molecules, known to be potent
differentiation inducers, showed similar effects to those of DMSO. The
potentiating effect of DMSO was related neither to an up-regulation of TNF
receptor expression nor to an alteration in the rate of TNF internalization
and degradation. We present evidence that the TNF activities are p55 TNF
receptor-mediated and are not due to insertion of TNF into lipid bilayers,
an effect that could be susceptible to DMSO, as this component has been
described to modify cell membrane characteristics. DMSO-induced
potentiation of TNF cytostasis/cytotoxicity was restricted to myeloid
leukemia cell lines. In non-myeloid cells such as fibrosarcomas,
myosarcomas, thymomas, or carcinomas, DMSO was found either not to alter or
to inhibit TNF-induced cell death. The latter results are in good agreement
with data reported by others who suggested that DMSO could act as a
scavenger of TNF-induced toxic radical formation. The potential correlation
in myeloid cells between DMSO-induced changes in the cells' differentiation
status and DMSO-enhanced TNF-susceptibility is discussed.

PMID: 7829967, UI: 95131071

Transfusion 1994 Oct;34(10):887-90

Hematopoietic progenitor cells are resistant to dimethyl sulfoxide

Branch DR, Calderwood S, Cecutti MA, Herst R, Solh H

Canadian Red Cross Society Blood Services, Toronto, Ontario.

BACKGROUND: A direct chemical toxicity of dimethyl sulfoxide (DMSO) to
hematopoietic progenitor cells has been suggested. However, a recent study
failed to corroborate these earlier findings. Thus, a series of experiments
was undertaken to address this issue. STUDY DESIGN AND METHODS: Bone marrow
was collected from 18 donors and cryopreserved with 10 percent (vol/vol)
DMSO. Aliquots of frozen bone marrow were thawed, diluted with ACD-A to 8
percent (vol/vol) DMSO, and allowed to remain in DMSO for up to 2 hours
before mononuclear cells were plated for colony-forming assays. After 14
days in culture, burst-forming units-erythroid, colony-forming
units--granulocyte/macrophage, and colony-forming
units--granulocyte/erythrocyte/macrophage/megakaryocyte colonies were
enumerated. RESULTS: There was no significant difference (p > 0.5) seen in
colony formation over the 2-hour exposure to DMSO. CONCLUSION: These
results support and extend a previous study that bone marrow hematopoietic
progenitor cells, including burst-forming units--erythroid, colony-forming
units--granulocyte/macrophage, and colony-forming
units--granulocyte/erythrocyte/macrophage/megakaryocyte are resistant to
any toxic effects of 8- to 10-percent (vol/vol) DMSO during at least 2
hours of DMSO exposure.

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