REPRODUCTION

RESEARCH

Zinc as a non-hormonal contraceptive: an alternative to the

copper intrauterine device (IUD)
K N Shankie-Williams , L A Lindsay, C R Murphy and S N Dowland
School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, New South
Wales, Australia
Correspondence should be addressed to K N Shankie-Williams; Email: kirst​en.sh​ankie​-will​iams@​sydne​y.edu​.au

Abstract
In brief: The many side effects of current contraceptives leave a large proportion of women without adequate protection. This study
shows that zinc, a highly physiologically compatible metal, provides effective long-acting reversible contraception in rats,
without requiring the use of hormones.
Abstract: Long-acting and reversible contraceptives (LARC) are the most widely used form of female contraception worldwide;
however, they have significant side effects that often result in early removal. Most LARCs are hormonal, but the use of
exogenous hormones is not suitable for all women and causes side effects in many others. The copper IUD (CuIUD) is the
only non-hormonal LARC, but a large proportion of users suffer severe side effects. This study proposes the use of zinc as a
suitable alternative to the CuIUD. A rat intrauterine device (IUD) model was established to test the efficacy of a zinc IUD
(ZnIUD) against a CuIUD. The IUD was surgically implanted into one uterine horn while the other remained untreated. Both
the ZnIUD and CuIUD resulted in zero implantation sites which were significantly fewer compared to non-treated horns.
Histological assessment revealed damage and inflammation in the endometrium of CuIUD-treated horns but only minor
epithelial changes in ZnIUD-treated horns. This suggests ZnIUDs may not share the side effect profile of the CuIUD. To test
the long-term efficacy of the ZnIUD, rats had a ZnIUD surgically implanted into both horns and cohoused with males for 3
months. These rats mated regularly but did not get pregnant, confirming long-term effectiveness. Reversibility of the ZnIUD
was also established, as removal of the ZnIUD after 3 months resulted in no significant difference in the number of
implantation sites between treated and untreated horns. This study demonstrated the contraceptive efficacy of zinc and its
potential as a LARC.
Reproduction (2022) 164 135–142

Introduction
Intrauterine devices (IUDs) are the most commonly
used form of reversible contraception worldwide
(d’Arcangues 2007). Almost all long-acting reversible
contraceptives (LARCs) contain hormones as their
contraceptive agent, and many women are either
precluded from using these by a medical condition
or wish to avoid the many physical and physiological
side effects of exogenous hormones. Depression, mood
swings, irritability, anxiety, weight gain, migraines and
decreased libido are just some of the commonly reported
side effects experienced by hormonal contraceptive
users in numerous studies over the last few decades
(Stephenson et al. 1994, Robinson et al. 2004, Beksinska
et al. 2010, Zettermark et al. 2018, Guen et al. 2021).
These can lead to discontinuation or inconsistent use,
both increasing the chance of unplanned pregnancies.
Only one non-hormonal option currently exists, the
copper IUD (CuIUD), which is a medicated model
that when inserted into the uterus provides the user
up to 12 years of protection against pregnancy. This
long-term contraceptive comes with many practical
benefits distinguishing it from alternatives, such as the
contraceptive pill, including low cost, no potential for
patient error and no exogenous hormone induced side
effects. These characteristics provide the foundation for a
low risk and financially accessible contraceptive option
for women around the world. However, the CuIUD has
its own unique side effect profile causing more painful
periods for 38% of women (Hubacher et al. 2009).
This excessive pain and bleeding, both menstrual and
intermenstrual, has led to persistent rates of early removal
of around 25%. For this reason, it is not recommended
for women with moderate to heavy periods (Hubacher
et al. 2009). One in 10 women worldwide have an
unmet need for contraception, and side effects are a
primary reason women discontinue use, resulting in 121
million unintended pregnancies each year (Sedgh et al.
2016, Bearak et al. 2020, UNDESA 2020).

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136 K N Shankie-Williams and others
Side effects experienced by users of the CuIUD are a
direct result of the copper content of the device. Animal
models have shown that copper cytotoxicity causes
necrosis, erosion, stromal hypertrophy and metaplasia of
epithelial cells in mucosal surfaces in direct contact with
copper, resulting in polyps (Zipper et al. 1969, Chvapil
et al. 1978). Exposure to metallic copper also resulted
in an increase in lysosomes and swollen, vesiculated
mitochondria in vaginal epithelial cells and infiltration
of leukocytes into the underlying mucosa (Chvapil
et al. 1978). This is consistent with the morphological
changes observed in human endometrial biopsies taken
from women using the CuIUD (Misra et al. 1989).
Lymphocytic, monocytic and leukocytic infiltrations
have been found in the uterine epithelium and stroma
of CuIUD users, and a loss of epithelial integrity was
observed (González-Angulo & Aznar-Ramos 1976).
During early experiments exploring non-hormonal
IUD materials, a range of metals were trialled for
antifertility effects and tested in the rabbit uterus
by placing an IUD near the cervix (Zipper et al.
1969). Copper and zinc resulted in the highest anti-
fertility rates of all the metals studied. Although only
one implantation site was found with the zinc IUD
(ZnIUD), further investigation of the use of zinc was
largely abandoned. However, we suggest that the
slightly lower efficacy of zinc in this study may have
resulted from methodological limitations. Later studies
have shown that the two-cell embryo has a unique
sensitivity to Zn2+, impeding further development
(Andrews et al. 1991); therefore, we hypothesise that
this may be responsible for the contraceptive effect of
a ZnIUD. As such, the placement of the experimental
suitable for copper, but suboptimal for zinc − being
IUDs near the cervix of rabbits (Zipper et al. 1969) was
located distant from the site at which we now expect
it to act.
There has only been one study that previously
considered the reversibility of a ZnIUD (Medel et al.
1972). This model involved rats and rabbits being
mated 2–3 days after insertion of IUDs, which were
then removed on day 2 after mating, with pregnancy
outcomes evaluated on day 10 (Medel et al. 1972). This
work showed pregnancy was able to proceed in these
animals; however, this study design only evaluates the
Reproduction
 (2022) 164 135–142
impact of IUD removal in early pregnancy rather than
demonstrating reversibility of these contraceptives as
claimed. In addition, authors acknowledged that surgical
trauma to the uterus during pregnancy was expected to
contribute to the reported findings of decreased fertility
in the IUD-treated horns.
Although these earlier studies provided initial
evidence supportive of zinc as a contraceptive agent, true
efficacy and reversibility of a ZnIUD have not yet been
established. Therefore, the present study has developed
a rat model to examine the contraceptive efficacy of a
ZnIUD with consideration of its potential mechanism of
action. This article also examines the potential for zinc
to provide continued contraception over an extended
period of time and assesses the return to fertility after
such long-term use of a ZnIUD, to determine the true
reversibility of such a contraceptive. The impact of a
ZnIUD on the endometrium is also compared with that of
a CuIUD. This work aims to evaluate the potential utility
of a ZnIUD as a non-hormonal LARC, in comparison
with the only current option, the CuIUD.
Methods
Ethics approval
All procedures were approved by The University of Sydney
Animal Ethics Committee.
Animals and mating
In total, 23 nulliparous female Wistar rats were housed in
plastic cages under a 12h light:12h darkness cycle at 21°C. All
rats were between 10–12 weeks old at the start of the protocol
and weighed 280–300 g. Rats were fed and watered ad libitum.
The oestrous cycle of the rats was tracked by vaginal smearing
and analysis of the cellular composition (Long & Evans 1922).
Mating was also confirmed by smearing, the presence of
sperm indicating day 1 post-coitum (pc). Pseudopregnancy
was confirmed by the cellular composition of daily vaginal
smears (Long & Evans 1922).
Treatment groups
Rats were randomly allocated to three experimental groups (Fig.
1). In the Efficacy Group, rats had either a zinc (n = 5), copper
Figure 1 Flowchart of experimental treatment
groups. The Efficacy Group had either a zinc
(n = 5), copper (n = 6) or nylon IUD (n = 5)
inserted into one uterine horn. The Long-term
Group (n = 3) rats had ZnIUDs inserted into
both uterine horns for 3 months. The
Reversibility Group (n = 4) had a ZnIUD
inserted into one uterine horn which was
removed 3 months later. Animals were mated
and tissue was collected on day 9 pc.
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(n = 6) or nylon IUD (n = 5) inserted into one uterine horn,
were mated after recovery and tissue was collected on day 9
pc. In the Long-term Group (n = 3), rats had ZnIUDs inserted
into both uterine horns, allowed to recover for 1 week and
then co-housed with a male for 3 months. Rats were regularly
smeared to track the oestrous cycle, mating events and number
of days in pseudopregnancy. At the end of the 3-month period,
tissue was collected 9 days after mating. In the Reversibility
Group (n = 4), rats had a ZnIUD inserted into one uterine horn
which was removed 3 months later. After recovery, animals
were mated and tissue was collected on day 9 pc.
Surgical insertion and removal of IUDs
Insertion
ZnIUD and CuIUD were made from 1 cm of 99.994% pure
zinc and 99.999% pure copper wire (Alfa Aesar, MA, USA),
0.5 mm in diameter and were finished with a small loop
at one end. Rats received an intraperitoneal injection of
ketamine (75 mg/kg; TROY Laboratories, NSW, Australia) and
xylazine (10 mg/kg; Parrell Laboratories, NSW, Australia). The
IUDs were inserted into the uterine lumen through a small
incision in the uterine wall, 1 cm from the ovary. All IUDs
were secured in place with a nylon suture through the IUD
loop and uterine wall, closing the uterine wall incision. A
foreign body control was also performed by inserting a 1-cm
loop of nylon thread into the uterine horn to test the effect
of the presence of a foreign object in the uterine lumen. Rats
were allowed to recover for at least 1 week after surgery.
Removal
Removal surgery for animals in the Reversibility Group
occurred during dioestrus, at which time the uterus is small,
not oedematous and lacks prominent vasculature, allowing
easier removal and minimising tissue damage. Similar to
insertion surgery, rats received an intraperitoneal injection
of ketamine (75 mg/kg) and xylazine (10 mg/kg), the uterine
horn was exposed and the IUD was carefully removed. Rats
recovered for at least a week.
Tissue collection
Rats were anaesthetised with 20 mg/kg sodium
pentobarbitone (TROY Laboratories, NSW, Australia) and
dissected under deep anaesthesia before euthanasia. Both
uterine horns were excised, pinned onto dental wax and
photographed. The implantation sites were counted and
compared to the control horn to determine contraceptive
efficacy (Liechty et al. 2015). Uteri were cut into
approximately 5 mm blocks from the region of the horn
containing the IUD or inter-implantation sites in the control
(Christofferson & Nilsson 1988) and immediately fixed in
10% neutral buffered formalin (Fronine, NSW, Australia).
The tissue was then washed in distilled H2O and dehydrated
in 30, 50 and 70% ethanol for 30 min each. Samples were
then placed in the Excelsior ES tissue processor (Thermo
Scientific) to further dehydrate tissue in graded ethanol baths
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Zinc as a non-hormonal contraceptive 137
over 4.5 h at 40°C before clearing with two 45-min changes
of xylene and infiltration with paraffin (Thermo Fisher)
over 3 changes of 30 min at 60°C. Finally, specimens were
embedded using the Tissue-Tek TEC 5 tissue embedding
console system (Sakura FineTek, Torrance, CA, USA).
Sectioning and staining
Transverse sections (7 µm) were cut on an HM 325 Rotary
Microtome (Thermo Fisher Scientific), and sections were
floated on a water bath (40°C) and placed onto gelatin-chrom-
alum-coated glass slides. At least five sections were collected
per block and two blocks were used for each animal. Slides
were then dried overnight in a 40°C oven.
Sections were stained using a routine haematoxylin and
eosin protocol (Carleton et al. 1980). Images were captured
on an Olympus BX53 brightfield microscope (Olympus), using
an Olympus DP73 camera (Olympus) and Olympus CellSens
software (version 1.11).
Statistical analysis
In the Efficacy Group, changes in the number of implantation
sites in IUD-treated horns and contralateral non-treated horns
were analysed using two-way ANOVA, with IUD material
and uterine horn as factors. Tukey post hoc test for multiple
comparisons (reporting multiplicity-adjusted P values)
was then applied to determine which pairs of means were
significantly different. Differences were determined to be
statistically significant when P < 0.05. For the data obtained in
the Reversibility Group, a paired t-test was used to determine
if a statistical difference existed between the number of
implantation sites present in a horn that had a ZnIUD removed
of ≤0.05 was considered statistically significant. All statistical
after 3 months and a contralateral non-treated horn. A P value
analysis was performed using GraphPad Prism Software
(version 7.02; GraphPad Software, Inc), and bar graphs were
generated where values represent the mean + s.e.m.
Results
Copper and zinc IUDs prevent pregnancy
The contraceptive efficacy of copper and zinc was assessed
by counting the number of implantation sites in the IUD-
treated and untreated horns on day 9 pc in rats receiving
different IUD materials (Fig. 2). In rats receiving a nylon
IUD, implantation sites were evident in both the IUD horn
and untreated, control horn with no significant difference
in the number of implantation sites in control vs treated
horns (P = 0.1059) (Fig. 2). In rats with either a CuIUD
or ZnIUD, there were no implantation sites in uterine
horns containing an IUD. Implantation sites were evident
in the untreated uterine horns, resulting in a statistically
significant difference in the number of implantation sites
in treated compared to untreated horns in both CuIUD
(P < 0.0001) and ZnIUD (P < 0.0001) animals (Fig. 2).
Reproduction (2022) 164 135–142
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138 K N Shankie-Williams and others
Figure 2 Contraceptive efficacy of metal IUDs (Cu2+ + Zn2+) and a control
nylon IUD on day 9 pc. (A) Implantation sites (arrow heads) were visible
in both IUD and control horns of the nylon treated rats, as well as the
region containing the IUD (bracket). (B) CuIUD-treated horns did not
contain implantation sites while the non-treated control horn contained
multiple. (C) ZnIUD-treated horns did not contain implantation sites
while the non-treated control horn contained multiple. (D) There was no
significant difference in the number of implantation sites between
treated and control uterine horns in rats receiving a nylon IUD (n = 5).
No implantation sites were seen in the CuIUD-treated (n = 6) and
ZnIUD-treated (n = 5) uterine horns in any animals, while the respective
control horns contained implantation sites. (**** P < 0.0001).
Short-term exposure to metal IUDs impacts
endometrial histology
Endometrial histology was assessed after exposure to
each of the IUD materials. The endometrium of the
copper-treated uterine horn exhibited infiltration of
inflammatory cells into the stroma, including dense
clusters of leukocytes, including neutrophils (Tigner et al.
2021) and sites of epithelial damage in the region of the
IUD (Fig. 3A). Along areas of intact epithelium, the simple
columnar epithelial cells had undergone metaplasia with
a resulting saw-tooth appearance (Fig. 3B). The underlying
stroma, glands and myometrium of the uterus appeared
normal. The untreated uterine horn demonstrated normal
histological appearance of the endometrium with no
epithelial damage, inflammation or metaplasia. In the
ZnIUD-treated uterine horns, the epithelium remained
intact, with no regions of epithelial damage or exposure
of the stroma and no dense clusters of leukocytes
(Fig. 3C). The epithelium showed mild metaplasia
resulting in an uneven apical border (Fig. 3D). The
Reproduction
 (2022) 164 135–142
Figure 3 Uterine histology of horns containing metal IUDs (Cu2+ and
Zn2+) on day 9 pc. (A) Haematoxylin and eosin showing inflammation in
the underlying stroma and epithelial damage in the CuIUD-treated horn.
(B) Metaplasia (arrows) of the uterine epithelial cells is present in the
CuIUD-treated horn. (C) The ZnIUD-treated horn exhibits normal overall
uterine morphology. (D) Mild metaplasia of the uterine epithelial cells is
present in the ZnIUD-treated horn. L, lumen; S, stroma; E, epithelium
underlying stroma, glands and myometrium appeared
normal. The untreated uterine horn did not show signs
of damage, inflammation or metaplasia. Uterine horns
containing a nylon IUD exhibited no abnormalities
and displayed normal uterine morphology (Fig. 4A
and B). The glands and myometrium appeared normal.
The untreated horn demonstrated normal histological
appearance with no signs of damage, inflammation or
metaplasia (Fig. 4C and D).
Zinc IUD provides long-term contraception
The long-term contraceptive efficacy of a ZnIUD was
assessed by inserting IUDs in both uterine horns and
co-housing rats with males for 3 months. Daily vaginal
smears showed that the rats regularly mated but did not
get pregnant. Cervical stimulation during mating causes
rescue of the corpus luteum in rats resulting in rats spending
an average of 10 days in a pseudopregnant state before
returning to oestrus and mating again (Smith et al. 1975).
Despite rats mating at least 8 times each, there were no
pregnancies during the 3-month treatment period (Table 1).
The contraceptive effect of the zinc IUD is reversible
The reversibility of the method was investigated by
inserting a ZnIUD into one uterine horn, removing it 3
months later and then examining implantation success.
No significant difference in the number of implantation
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 Zinc as a non-hormonal contraceptive 139

Figure 5 Number of implantation sites in uterine horns after a

Figure 4 Uterine histology of horn containing nylon IUD and
contralateral non-treated horn on day 9 pc. (A) Haematoxylin and
eosin showing no abnormalities present in the nylon IUD-treated
horn. (D) The contralateral non-treated horn exhibits normal uterine
morphology. L, lumen; S, stroma; E, epithelium
sites in the ZnIUD-treated uterine horn compared to
the untreated horn was found (P = 0.86) (Fig. 5). The
average number of implantation sites in ZnIUD-treated
uterine horns was 5.75 (±1.4) compared to 6 (±0.4)
implantation sites in control horns.
3-month treatment and subsequent removal of a ZnIUD (n = 4). There
was no significant change in implantation rate between treated and
untreated horns after IUD removal from the treated horns.
100% contraceptive efficacy, completely preventing
pregnancy in the IUD-treated uterine horns. This study
also demonstrated for the first time that a ZnIUD can
provide 100% contraceptive efficacy, equal to that of the
CuIUD. Comparison with the nylon IUD demonstrated
that the contraceptive effect of both the ZnIUD and

Long-term exposure to the zinc IUD does not impact

endometrial histology
Histological analysis of the endometrium after 3 months
exposure to a ZnIUD revealed no pathologies such
as metaplasia or inflammation in the ZnIUD-treated uterus
(Fig. 6A and B). The untreated horn did not show signs of
damage, inflammation or metaplasia (Fig. 6C and D).

Discussion
Both copper and zinc demonstrate 100%
contraceptive efficacy in the rat IUD model
In agreement with previous animal studies, the
present study determined that the CuIUD provided

Table 1 Contraceptive efficacy of the ZnIUD over a 3-month period.

Mating Mating Mating pair

pair 1 pair 2 3
Figure 6 Uterine histology of horn containing a ZnIUD for 3 months
No. of matings 8 8 9 and the non-treated control horn after removal and mating. (A and B)
No. of pregnancies 0 0 0 Haematoxylin and eosin showing no abnormalities of the uterine
Average no. of days in 11 11 9 epithelial cells and underlying stroma in the ZnIUD-treated horn,
pseudopregnancy closely resembling that of the non-treated control horn (C and D). L,
Average no. of days in oestrus 1 1 1
lumen; S, stroma; E, epithelium

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140 K N Shankie-Williams and others
CuIUD is due to the metal content of the IUD, rather
than resulting from the presence of a foreign body in
the uterine lumen. Our IUD model demonstrated higher
contraceptive efficacy than Zipper et al. (1969), showing
this model successfully takes into consideration the
mechanism of action.
Zinc IUD is an effective long-term and reversible
contraceptive in the rat
In order to mimic the clinical setting, this study for the
first time showed the long-term contraceptive efficacy
and reversibility of a ZnIUD. The ZnIUD prevented
all pregnancies in female rats co-housed with males
for a continuous 3-month period with no change in
the number of days in oestrus or the average days of
pseudopregnancy after cervical stimulation (Long &
Evans 1922). This indicates that zinc may have long-term
efficacy in clinical settings similar to that of the CuIUD.
The present study is also the first to clearly demonstrate
that the contraceptive effect of a ZnIUD is reversible.
When ZnIUDs were removed after 3 months exposure,
rats were able to rapidly return to fertility and exhibited
successful implantation in uterine horns previously
containing an IUD, at rates comparable to untreated
uterine horns. This is in contrast to the previous study
which found a significant decrease in implantation sites
when ZnIUDs were removed 2 days after mating, in
treated compared to control horns (Medel et al. 1972).
As such, this study has shown that the ZnIUD could be
an effective LARC and offer an alternative contraceptive
option to women.
Copper IUD impacts endometrial histology more
severely than zinc
The morphological changes and pathologies of the
CuIUD-treated horn, observed histologically in the
present study, reflect those seen in the earlier rabbit and
human trial of the CuIUD (Zipper et al. 1969). The CuIUD
in our rat IUD model caused severe metaplasia in the
uterine epithelium and epithelial damage. Inflammation
was also present in the CuIUD horn, in alignment
with the stromal hypertrophy documented by Zipper
et al. (1969) in copper-treated rabbit uterus (Zipper
et al. 1969). An influx of inflammatory cells around
areas that had lost epithelial integrity were common
amongst the copper-treated horns. This inflammation
and endometrial damage may lead to many commonly
reported side effects. Inflammatory mediators activate
and sensitise local nociceptors to prevent further tissue
damage (Julius & Basbaum 2001), whilst the continuous
release of proteolytic agents from leukocytes suppress
endometrial angiogenesis, resulting in pain and excess
bleeding (Ferenczy 2003). Recent studies have found a
higher concentration of copper in the endometrium of
women experiencing abnormal bleeding with a CuIUD
Reproduction
 (2022) 164 135–142
and a positive correlation with vascular endothelial
growth factor (VEGF), a potent angiogenic factor (Li
et al. 2016). In addition, higher fibrinolytic activity
has been found in areas of the endometrium that have
direct contact to the CuIUD, indicating inhibition of
homeostasis (Liedholm et al. 1983). The histological
data in the present study support these observations,
further supporting that copper directly exerts negative
effects on the endometrium.
These effects are particularly severe soon after
insertion, when the rate of dissolution of copper ions
into the uterine fluid is highest upon insertion of the
CuIUD and drops exponentially over the duration of
its use, an effect known as the ‘burst release’ (Alvarez
et al. 2012). This high initial Cu corrosion rate irritates
the endometrium, enhancing the inflammatory
response and deposition of proteins and cell debris
and may contribute to the inflammation and damage
of the endometrium after short-term exposure to the
CuIUD, seen in this study. This period is when the
highest removal rates occur, preventing many women
from having successful, long-term use of the CuIUD
(Hubacher et al. 2009).
The impact of a ZnIUD on uterine histology has
not been previously examined. Short-term exposure
(approximately 2–3 weeks; Efficacy Group) to the
ZnIUD resulted in mild metaplasia in the uterine
horn, with epithelial cells appearing irregular in
height. Unlike the CuIUD-treated horn, the epithelium
remained intact and no areas of inflammation were
observed. Therefore, it is possible that a ZnIUD would
cause users less pain and bleeding compared to the
CuIUD, due to the absence of localised inflammation
(Julius & Basbaum 2001, Ferenczy 2003). Uterine
horns that were subjected to long-term exposure
to Zn (approximately 16 weeks; Long-Term Group)
exhibited no pathologies, having the same appearance
as the control horn. This indicates that Zn acts as
a contraceptive without any long-term impact to
endometrial integrity. Although there have been
significantly fewer studies conducted on the interaction
of zinc in the uterus compared to those of copper, the
present findings, in conjunction with the emerging use
of zinc in biocompatible implants provide a positive
indication of the potential biocompatibility of a ZnIUD
(Drelich et al. 2017, Kannan et al. 2017). Zinc is
relatively harmless to humans as it is the second most
abundant essential metal in the body and found at a
mean serum concentration of 1000 μg/mL in humans,
with a recommended daily intake of 10–15 mg (Plum
et al. 2010). This is in contrast to copper, which is
found at a mean serum concentration of 0.1 μg/mL
and a recommended daily intake of 0.9 mg. Because
of this, the body is physiologically adept at transporting
and storing larger concentrations of zinc with no risk of
producing reactive oxygen species, a consequence of
high concentrations of copper and mediators of acute
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and chronic inflammatory pain (Khattab 2006, Babula
et al. 2012). Zn is now recognised as an excellent
candidate for Zn-based orthopaedic implants, being
chosen for its biocompatibility and safety (Kannan
et al. 2017, Yang et al. 2020).
Contraceptive effect of a zinc IUD is likely via a
unique mechanism, acting on the two-cell embryo
The improved contraceptive efficacy of zinc in the
present study compared to earlier studies (Zipper et al.
1969, Chang et al. 1970, Kincl & Rudel 1971) supports
our hypothesis that Zn exerts its contraceptive effect via a
unique effect on the two-cell embryo. Previous research
has shown that the two-cell embryo exhibits specific
vulnerability to Zn2+ (Andrews et al. 1991). At this stage,
the embryo has begun transcription of metallothionein,
an important metal binding protein. However, at such
an early stage, metallothionein is a precursor protein
without metal reactivity (Andrews et al. 1991). The metal
responsiveness occurs later in development meaning the
presence of the slightly increased concentrations of Zn2+
during this window of development may be lethal to the
embryo. The location of the ZnIUD prior to fertilisation
in our model facilitates high contraceptive efficacy and
would explain the lack of a contraceptive effect seen
in earlier studies when a Zn wire was inserted 3 days
post fertilisation (Chang et al. 1970), when embryos
would already have progressed past the two-cell stage.
The present study also demonstrates higher contraceptive
efficacy than another model used by Kincl and Rudel
(Kincl & Rudel 1971), where a 3-mm Zn loop was inserted
into the uterus 3 days post copulation. This supports our
suggestion that zinc acts on the two-cell embryo, which
is uniquely vulnerable to Zn (Andrews et al. 1991),
such that the ZnIUD must be in place within 24 h post-
fertilisation, before the zygote cleaves into 2 cells.
This study has refined an animal model to study
the effectiveness of non-hormonal IUDs in the rodent
uterus and for the first time allowed direct evaluation of
the endometrial responses to these metals. This model
showed that a ZnIUD provides long-term and reversible
contraception without causing local tissue damage to the
endometrium. As such the ZnIUD stands up to the gold-
standard for non-hormonal contraception, the CuIUD.
The ZnIUD may also have an additional advantage of
a new mechanism of action that impacts the embryo
directly, avoiding irritation and inflammation of the
uterus. Together, these findings suggest that the ZnIUD
may have utility as a LARC that could have significantly
fewer side effects.
Declaration of interest
The authors declare that there is no conflict of interest that
could be perceived as prejudicing the impartiality of the
research reported.
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Zinc as a non-hormonal contraceptive 141
Funding
This work was supported by an NWG Macintosh Memorial
Grant from The Ann Macintosh Foundation of the Discipline of
Anatomy and Histology and Murphy Laboratory Funds.
Author contribution statement
K N S-W designed the study, carried out experimental work,
analysed data and prepared manuscript. L A L and C R M gave
technical advice as well as experimental design and editing
of manuscript. S N D designed the study, gave technical and
experimental assistance and critical editing of manuscript.
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Received 1 May 2022
First decision 8 June 2022
Revised manuscript received 10 July 2022
Accepted 5 August 2022
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