166 Full

166
ANE5TH ANALG
1988:67 166-9
Reproductive and Teratogenic Effects of Sufentanil and Alfentanil

in Sprague-Dawley Rats
Masahiko Fujinaga, MD, Richard I. Mazze,
Jeffrey M. Baden, MD
MD,
Ethan C. Jackson, and
FUJINAGA M, MAZZE R1, JACKSON EC, BADEN JM

Reproducti\re and terafogenic effects ot wfentanil aiid
altentanil in Sprague-Dawlev Rat5 Anesth Analg
1988.67 1 6 6 9
Key Words: NARCOTICS-sufentanil, alfentanil.

PREGNANCY, TEiiAToCENIcITY-sufentanil,
alfentanil. TOXICITY, REPRODUCTIVE-SUfentanil,
alfentanil.
For the past 10 years we have studied the reproductive and teratogenic effects of the inhaled anesthetics
in several rodent models (1-8). Until recently, reliable
studies of parenteral anesthetic agents and narcotics
were not possible because of the extensive maternal
toxicity associated with bolus administration of even
relatively low doses of these drugs. The use of
chronically implanted subcutaneous (SC) osmotic
minipumps has, however, permitted delivery of high
total daily doses of local anesthetics and narcotics a t a
constant rate throughout organogenesis without causing adverse maternal physiologic effects. Thus, we
have studied lidocaine and fentanyl and found that
they are devoid of adverse reproductive effects (9,lO).
In the present study, we have extended our reproduction and teratology investigations of parenteral agents
to include the new narcotics, sufentanil and alfentanil.
Methods
Timed pregnant Sprague-Dawley rats were received
from the breeder (Hilltop Lab Animals, Inc., ScottSuppcirted by the Veterans Administration and the Anestliesia
Phdrmacology Rt,search Foundation. Received frorn thc Dcpartmeiit of Anesthesia, Stantorcl University School of hledicine,
Stantord, California, aiid the Aiiesthe~iologyService, Palo Alto
Veterans Aciministr,ition Medical Center, Palo Alto, CLjlitornia.
Accepted for publication September 21, 1987.
dale, PA) either on day 2 (sufentanil experiment) or

on day 4 of pregnancy (alfentanil experiment),
weighed, and identified with metal ear tags. (Day 0 of
pregnancy was defined as the day a copulatory plug
was observed in the vagina.) Rats were bedded on
ground corncob (Bed-OCobs, Andersons Cob Division, Maumee, OH), housed four per cage and fed
standard laboratory rodent food (Wayne Lab Blox,
Allied Mills, Inc., Chicago, IL) and tap water ad
libitum. Temperature i n the animal room was maintained a t 21-24C and artificial light was provided
from 6 AM to 7 PM each day. Care of rats was in
accordance with institutional guidelines.
Two separate experiments of similar design were
performed. In each experiment, on day 5 of pregnancy, an osmotic minipump with a 15-day drug
supply (Alzet miniosmotic pump, model 2ML2, 5
plihr delivery rate, Alza Corp., Palo Alto, CA) was
implanted SC in the back of each rat (11). To accomplish the procedure, which took less than 1 minute to
complete, general anesthesia was induced with an IM
injection of 0.3 mlikg of a mixture containing ketamine 60 mgiml, xylazine 6 mgiml, and acepromazine,
1.2 mgiml. In the first experiment, 128 rats were
assigned to four groups of approximately equal average weight and were treated with either: 1) saline
(control; t i = 39); 2) a low dose of sufentanil (10
pg.kg-.day-; 11 = 30); 3) an intermediate dose of
SUFENTANIL A N D ALFENTANIL TERATOGENICITY
Table 1. Summary of Group Body Weight and Body Weight Changes*

~
Alfentanil
(nigikg
pcr day
Sufentanil (@kg per day)
Group
Control
in
5n
100
Control
No. of rats studied

Body weight
On arrival from breeder
Day 11 of pregnancy
Day 20 of pregnancy
(day of cesarean
section)
39
30
30
29
20
20
202 t 16
244 2 15
312 2 28
202 t 14
245 t 14
347 2 23
203 t 16
242 t 17
332 -+ 4n
201 2 14
241 2 14
327 ? 38
204 t- 14
265 2 16
359 2 23
207 t 12
268 ? 19
34.5 t 32
*Values are in grams (mean
SD)
sufentanil (50 pg.kg-l.day-'; IZ = 30); or 4) a high

dose of sufentanil (100 pg.kg-'.day-'; n = 29). In the
second experiment, 40 rats were allocated to two
groups and were treated with either; 1) saline (control; I I = 20); or 2) a high dose of alfentanil (8
mg.kg-'.day-'; n = 20). Sufentanil and alfentanil
solutions were prepared from citrate powders
(Janssen Pharmaceutica, Piscataway, NJ) dissolved in
normal saline. Dosages of the narcotics were based
on data from preliminary studies in which we determined the highest dose that did not cause maternal
weight loss. In our experience with narcotics delivered with osmotic minipumps, maternal weight loss
consistently occurs before respiratory depression.
Only the highest dose of alfentanil was studied with
the intention of studying lower doses if the single
dose gave positive results. Rats were weighed every
2-3 days during the experiment.
On day 20 of pregnancy, 1day before the expected
delivery date, rats were killed by carbon dioxide
inhalation and cesarean section was performed. The
uterus was examined and the number and position of
live and dead fetuses, resorptions, and implantations
were recorded. The weight and sex of each live fetus
were determined and all fetuses were examined for
external abnormalities. Every other fetus was fixed in
70% ethanol and macerated with potassium hydroxide. The skeleton was then stained with alizarin red S
using the modified method of Staples and Schnell
(12), cleared with glycerol, and subsequently examined microscopically for skeletal abnormalities. The
remainder of the fetuses were preserved in Bouin's
solution and subsequently dissected and examined
microscopically for visceral abnormalities as described by Barrow and Taylor (13). All examinations
were done without knowledge of the treatment
groups.
Abnormalities were classified as follows. Fetal
morphologic abnormalities that alter general body
conformation, disrupt or interfere with bodily func-
tions, or generally appeared to be incompatible with

life were categorized as major malformations. Abnormalities in anatomic structure that were considered to
have no significant biological effect on animal health
or body conformity and represented only slight deviations from normal were categorized as developmental variants. Abnormalities that did not fall under the
strict definition of major malformations but which
were clearly not developmental variants were categorized as minor anomalies.
Stat is t ical Analysis

The percentage of abnormal fetuses in each litter of
each treatment group was computed and group
means were compared with control group means by
one-way analysis of variance (ANOVA). Student's t
test, corrected for multiple comparisons (Bonfferoni),
was used as an a posteriori test when differences
were found with ANOVA. P < 0.05 was considered
statistically significant.
Results
Materiinl Effects
No rats treated with sufentanil died during the experiment. Four rats treated with alfentanil died, one
on each of the first 4 days after pump implantation.
Otherwise, no adverse effects were observed during
the experiments. There were no differences in weight
gain among any of the groups in either experiment
(Table 1).
Rcyroductiue Efticfs
There were no significant differences among the
groups in pregnancy rate, total numbor of implanta-
168
FUJINAGA ET At..
ANESTH ANALG
1988;6716&9
Table 2. Maternal and Fetal Observations at Ccsarcan Section on Day 20 of Pregnancy*

Alfentanil
( m g h per
day)
Sufentanil ( p g i k g per day)

Group
Control
10
50
100
Control
No. of rats studied

No. of rats pregnant
Pregnancy rate ("i)
No. of implantationdrat
No. of live fetusesirat
Percent fetal wastagdrat ( % )
Percent femaleirat ( %)
Mean fetal iueight (g)
39
34
87
30
30
2 3
73
12.0 -c 2.0
11.1 ? 1.8
5.1 2 7.0
52.1 % 13.1
1.7 2 0.3
29
26t
90
11.5 i 2.0
10.7 t 2.4
7.1 t- '1.2
49 1 t- 14.8
4.7 2 0.3
20
18
90
20$
11
69
11.3 ? 2.9
9.8 i- 4.2
11.5 2 24.8
60.05 t- 19.5
4.3 2 0.3
11.9 t
11 6 2
2.5 t
493 2
4.5 2
25
2.4
2.4
4.5
110
0.2
83
12.0 t 2.0
11.8 t- 1.9
1 6 ? 3.9
33 2 ? 15.1
4.6 2 0.2
11.7 2 2.0
11.2 i- 2.0
3.7 2 5.5
4.1 1 t- 17.7
4.6 2 0.5
Table 3 . S u m m a r y of Fetal Examination

Alfen tanil
(mgikg per
day)
Sufentanil (p*!kgiday)
Group
Control
10
50
100
No. of rats examined

External examinations
No. of fetuses examined
Any abnormalities ( % )
Runt ([A)
Visceral examinations
Major malformations ( % )
Minor anomalies (<k)
Skeletal examinations
Major malformations ( U )
Minor anomalies ( % )
Developmental variant5 ( % )
34
25
20
24
18
227
0.0 2 0.0
0.0 f 0.0
257
0.0 t 0.0
0.3 5 1 . 4
202
0.0 i- 0.0
0.6 t 2.4
108
0.0 t 0.0
3.5 t 6.6
Control
8
11
?Y4
296
0.0 2 0.0
0.2 L 1.4
0.0 L O . u
0.4 t 2.0
198
0.0 t- 0.u
7.9 f 13.5
0.0 ? 0.0
2.2 i- 6.0
112
0.0 t 0.0
5.0 2 7.8
130
0.0 0.0
3.9 t 7.9
100
0.0 t 0.0
5.2 t 8.9
53
0.0 t 0.0
3.1 t- 6.6
196
0.0 -t 0.0
0.0 2 0.0
25.5 2 23.5
148
0.0 L 0. (1
1.1 t 3.9
34.0 L 32.0
115
0.0 f 0.0
0.0 t 0.0
32.3 t 28.3
127
0.0 t 0.0
0.0 t 0.0
30.5 2 27.7
102
0.0 f 0.0
0.9 t 4.0
16.8 t- 22.6
55
0.0 t 0.0
0.0 t 0.0
33.9 2 31.8
Mean percentage abnormal ictuses per rat
I48
I
\D
tions and live fetuses, percent fetal wastage per litter,

or mean fetal weight (Table 2). There were more
female offspring in the alfentanil group than in the
control group, the difference achieving statistical significance in part due to the lower than usual percentage of male offspring in the control group.
Discussion
In the present study, we tested the reproductive and
teratogenic effects of the narcotics, sufentanil and
alfentanil, and found both agents to be benign. In this
respect, our results are s i m i l a r to those we obtained
with the parent compound in this series, fentanyl
(10). We found that fentanyl in doses u p to 500
pg.kg '.day I , delivered with osmotic minipumps
for 2 weeks before breeding and during the entire
period of pregnancy, was devoid o f adverse reproductive effects. In another study, adding 500
pg.kg-'.day-' of fentanyl to 50% N,O did not increase the teratogenic effects of 50% N,O alone (14).
However, our results differ from those of studies of
the older narcotics, morphine, meperidine and methadone, in which adverse reproductive effects gener~
T mlogeti ic Effccts
A total of 1174 offspring were delivered from the 107
pregnant rats and all were examined for external
abnormalities. Subsequently, 588 fetuses were examined for visceral and 586 for skeletal abnormalities.
The results of these examinations are shown in Table
3 . There were no significant teratologic findings in
any group.
ANESTH ANALG
1988;67.16&9
SUFENTANIL AND ALFENTANlL TERATOGENICITY
ally were reported at high doses (15-19). In these

other investigations, narcotics were given once or
twice daily by various parenteral routes. From the
doses and side effects reported in these other studies,
and from our own studies (lo), we surmise that the
test animals experienced respiratory depression.
Thus, it is difficult to compare our results with the
new narcotics with those of past studies because the
design of the older studies almost certainly was flawed.
Apart from avoiding respiratory depression and
the thoracic rigidity associated with administration of
high doses of narcotics, there are other advantages to
using osmotic pumps for reproduction and teratology
studies. It is common in studies such as the present
one to administer the test drug throughout the period
of organogenesis, days 6 1 5 of gestation in the rat.
For agents that cannot be administered orally or by
inhalation, this necessitates daily injections. The handling and trauma associated with multiple injections
can in themselves be confounding experimental factors. Another problem with administration of bolus
doses of narcotics is that, with a relatively short
half-life, withdrawal may occur between doses. In
fact, Lichtblau and Sparber (20) have suggested that
in utero withdrawal from narcotics was more detrimental to the rat fetus than was continued exposure.
By using chronically implanted SC osmotic minipumps,
constant plasma levels of test agent can be maintained
from day 5, the day before implantation, until day 20,
the day before parturition, when rats are killed.
In terms of experimental design, the cost of using
SC-implanted osmotic minipumps is small. Although
general anesthesia is required to insert the pumps,
the procedure is quick and relatively noninvasive.
Also, Nau et al. (21) have reported that the total daily
dose of a given agent (valproic acid) required to
produce embryotoxicity was approximately ten times
higher when it was delivered with SC-implanted
osmotic pumps than by injection. Assuming the same
ratio applies to narcotics, the amount of narcotic that
can be delivered with osmotic minipumps without
causing adverse maternal effects is still far in excess of
the amount that can be safely delivered by injection,
i.e., approximately 100:l for fentanyl (10).
In conclusion, sufentanil and alfentanil do not
cause adverse reproductive or teratogenic effects in
rats when administered throughout organogenesis
with SC-implanted osmotic minipumps. From a methodologic point of view, osmotic minipumps facilitate study of the reproductive and teratogenic effects
of narcotics as they permit delivery of dosages that
would not be tolerated by bolus injection or might
otherwise confound experimental design.
169
References
1. Wharton RS, Mazze R1, Baden JM, Hitt BA, Dooley JR.
Fertility, reproduction, and postnatal survival in mice chronically exposed to halothane. Anesthesiology 1978;48:167-71.
2. Wharton RS, Wilson AI, Mazze RI, Baden JM, Rice SA. Fetal
morphology in mice exposed to halothane. Anesthesiology
1979;51:532-7.
3. Wharton RS, Sievenpiper TS, Mazze RI. Developmental toxicity of methoxyflurane in mice. Anesth Analg 1980;59:421-5.
4. Wharton RS, Mazze RI, Wilson AI. Reproduction and fetal
development in mice chronically exposed to enflurane. Anesthesiology 1981;54:505-10.
5. Mazze RI, Wilson AI, h c e SA, Baden JM. Reproduction and
fetal development in mice chronically exposed to nitrous
oxide. Teratology 1982;26:11-6.
6. Mazze RI, Wilson AI, Rice SA, Baden JM. Reproduction and
fetal development in rats exposed to nitrous oxide. Teratology
1984;30:259-65.
7. Mazze RI, Wilson AI, Rice SA, Baden JM. Fetal development in
mice exposed to isoflurane. Teratology 1985;32:33945.
8. Mazze RI, Fujinaga M, Rice SA, Harris SB, Baden JM. Reproductive and teratogenic effects of nitrous oxide, halothane,
isoflurane, and enflurane in Sprague-Dawley rats. Anesthesiology 1986;64:33944.
9. Fujinaga M, Mazze RI. Reproductive and teratogenic effects ot
lidocaine i n S p r a g u e - D a w l e y r a t s . A n e s t h e s i o l o g y
1986;65:62&32.
10. Fujinaga M, Stevenson JB, Mazze RI. Reproductive and teratogenic effects of fentanyl in Sprague-Dawley rats. Teratology
1986;34:51-7.
11. Theeuwes F, Yum SI. Principles of the design and operation of
generic osmotic pumps for the delivery of semisolid or liquid
drug formations. Ann Biomed Eng 1976;4:34>53.
12. Staples RE, Schnell VL. Refinements in rapid clearing technique in the KOH-alizarin red S method for fetal bone. Stain
Techno1 1968;39:61-3.
13. Barrow MV, Taylor WJ. A rapid method for detecting malformations in rat fetuses. J Morphol 1967;127:291-306.
14. Mazze RI, Fujinaga M, Baden JM. Reproductive and teratogenic
effects of nitrous oxide, fentanyl and their combination in
Sprague-Dawley rats. Br J Anaesth 1987;59:1291-7.
15. Harpel HS, Gautieri RF. Morphine-induced fetal malformations. J Pharm Sci 1968;57:1590-7.
16. Zagon IS, McLauglilin PJ. Effects of chronic morphine admin-
istration on pregnant rats and their offspring. Pharmacology

1977;15:302-10.
17. Hutchings DE, Hunt HF, Towey JP, Rosen TS, Gorinson HS.
Methadone during pregnancy in the rat: dose level effects on
maternal and perinatal mortality and growth in the offspring.
J Pharmacol Exp Ther 1976;197:171-9.
18. Geber WF, Schranim LC. Congenital malformations of the
central nervous system produced by narcotic and analgesics in
the hamster. Am J Obstet Gynecol 1975;123:705-13.
19. Jurand A. Teratogenic activity of methadone hydrochloride in
mouse a n d chick embryos. J Embryo1 Exp Morphol
1973;30:449-58.
20. Lichtblau L, Sparber SB. Opiate withdrawal in utero increase

neonatal morbidity in the rat. Science 1981;212:94>5.
21. Nau H, Zierer R, Spielmann H, Neubert D, Gansau C. A new
model for embryotoxicity testing: teratogenicity and pharmacokinetics of valproic acid following constant-rate administration in the mouse using human therapeutic drug and metabolite concentrations. Life Sci 1981;29:2803-1.

166 Full

Uploaded by

Copyright:

Available Formats

166 Full

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

166 Full

Uploaded by

Copyright:

Available Formats

166

Reproductive and Teratogenic Effects of Sufentanil and Alfentanil

Ethan C. Jackson, and

FUJINAGA M, MAZZE R1, JACKSON EC, BADEN JM

Key Words: NARCOTICS-sufentanil, alfentanil.

dale, PA) either on day 2 (sufentanil experiment) or

SUFENTANIL A N D ALFENTANIL TERATOGENICITY

Table 1. Summary of Group Body Weight and Body Weight Changes*

Sufentanil (@kg per day)

No. of rats studied

*Values are in grams (mean

sufentanil (50 pg.kg-l.day-'; IZ = 30); or 4) a high

tions, or generally appeared to be incompatible with

Stat is t ical Analysis

Table 2. Maternal and Fetal Observations at Ccsarcan Section on Day 20 of Pregnancy*

Sufentanil ( p g i k g per day)

No. of rats studied

Table 3 . S u m m a r y of Fetal Examination

No. of rats examined

Mean percentage abnormal ictuses per rat

tions and live fetuses, percent fetal wastage per litter,

SUFENTANIL AND ALFENTANlL TERATOGENICITY

ally were reported at high doses (15-19). In these

istration on pregnant rats and their offspring. Pharmacology

20. Lichtblau L, Sparber SB. Opiate withdrawal in utero increase

You might also like