Methods and Model Systems Used to Study Pregnant Human Uterine Smooth Muscle

Ilicic, Marina; Paul, Jay P.

doi:10.5772/intechopen.75201

Cited by 4 publications

(3 citation statements)

References 89 publications

(214 reference statements)

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“…Based on the in vitro inhibition of Ca 2+ -mobilization from the myometrial cells and in vitro uterine-selectivity compared to aortic VSMCs by mundulone and MA, we chose to examine their tocolytic ability and confirm uterine-selectivity at the tissue level. A well-established and widely utilized ex vivo isometric contractility assay was used to examine the effect of mundulone and MA on contractile activity of term pregnant mouse myometrial tissue [3,[33][34][35][36][37][38][39]. During our initial experiments, we observed the precipitation of mundulone and MA from a DMSO stock solution at higher concentrations tested in an aqueous organ bath assay (data not shown).…”

Section: Ex Vivo Tocolytic Ability and Lack Of Off-target Effect On Fetal Ductus Arteriosusmentioning

confidence: 89%

Identification of mundulone and mundulone acetate as natural products with tocolytic efficacy in mono- and combination-therapy with current tocolytics

Siricilla

et al. 2021

Preprint

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Currently, there are a lack of FDA-approved tocolytics for the management of preterm labor. We previously observed that the isoflavones mundulone and mundulone acetate (MA) inhibit intracellular Ca2+-regulated myometrial contractility. Here, we further probed the potential of these natural products to be small molecule leads for discovery of novel tocolytics by: (1) examining uterine-selectivity by comparing concentration-response between human primary myometrial cells and a major off-target site, aortic vascular smooth muscle cells (VSMCs), (2) identifying synergistic combinations with current clinical tocolytics to increase efficacy or and reduce off-target side effects, (3) determining cytotoxic effects and (4) investigating the efficacy, potency and tissue-selectivity between myometrial contractility and constriction of fetal ductus arteriosus (DA), a major off-target of current tocolytics. Mundulone displayed significantly greater efficacy (Emax = 80.5% vs. 44.5%, p=0.0005) and potency (IC50 = 27 μM and 14 μM, p=0.007) compared to MA in the inhibition of intracellular-Ca2+ from myometrial cells. MA showed greater uterine-selectivity, compared to mundulone, based on greater differences in the IC50 (4.3 vs. 2.3 fold) and Emax (70% vs. 0%) between myometrial cells compared to aorta VSMCs. Moreover, MA demonstrated a favorable in vitro therapeutic index of 8.8, compared to TI = 0.8 of mundulone, due to its significantly (p<0.0005) smaller effect on the viability of myometrial (hTERT-HM), liver (HepG2) and kidney (RPTEC) cells. However, mundulone exhibited synergism with two current tocolytics (atosiban and nifedipine), while MA only displayed synergistic efficacy with only nifedipine. Of these synergistic combinations, only mundulone + atosiban demonstrated a favorable TI = 10 compared to TI=0.8 for mundulone alone. While only mundulone showed concentration-dependent inhibition of ex vivo mouse myometrial contractions, neither mundulone or MA affected mouse fetal DA vasoreactivity. The combination of mundulone and atosiban yielded greater tocolytic efficacy and potency on term pregnant mouse and human myometrial tissue compared to single-drugs. Collectively, these data highlight the difference in uterine‐selectivity of Ca2+‐mobilization, effects on cell viability and tocolytic efficacy between mundulone and MA. These natural products could benefit from medicinal chemistry efforts to study the structural activity relationship for further development into a promising single- and/or combination-tocolytic therapy for management of preterm labor.

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Section: Ex Vivo Tocolytic Ability and Lack Of Off-target Effect On Fetal Ductus Arteriosusmentioning

confidence: 89%

Identification of mundulone and mundulone acetate as natural products with tocolytic efficacy in mono- and combination-therapy with current tocolytics

Siricilla

et al. 2021

Preprint

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“…The remaining tissue is stored at 4 °C and used within one 51 or two 52 days of collection. Samples are then further sliced into tissue strips on the order of millimeters, 37,53 often using an oscillating vibroslicer. 49 The length of the strip ultimately depends on the type of analysis that will be performed, whether functional or molecular.…”

Section: In Vivo and Ex Vivo Models Of The Myometriummentioning

confidence: 99%

Tools, techniques, and future opportunities for characterizing the mechanobiology of uterine myometrium

Maxey

McCain

2021

Exp Biol Med (Maywood)

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The myometrium is the smooth muscle layer of the uterus that generates the contractions that drive processes such as menstruation and childbirth. Aberrant contractions of the myometrium can result in preterm birth, insufficient progression of labor, or other difficulties that can lead to maternal or fetal complications or even death. To investigate the underlying mechanisms of these conditions, the most common model systems have conventionally been animal models and human tissue strips, which have limitations mostly related to relevance and scalability, respectively. Myometrial smooth muscle cells have also been isolated from patient biopsies and cultured in vitro as a more controlled experimental system. However, in vitro approaches have focused primarily on measuring the effects of biochemical stimuli and neglected biomechanical stimuli, despite the extensive evidence indicating that remodeling of tissue rigidity or excessive strain is associated with uterine disorders. In this review, we first describe the existing approaches for modeling human myometrium with animal models and human tissue strips and compare their advantages and disadvantages. Next, we introduce existing in vitro techniques and assays for assessing contractility and summarize their applications in elucidating the role of biochemical or biomechanical stimuli on human myometrium. Finally, we conclude by proposing the translation of “organ on chip” approaches to myometrial smooth muscle cells as new paradigms for establishing their fundamental mechanobiology and to serve as next-generation platforms for drug development.

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“…A study by Nani [4] resulted that drinking the soaking water containing a dose of 40 grams of A. hierochuntica L. increased the frequency of uterine contractions [4]. However, until now, the mechanism that regulates myometrial contraction activity during pregnancy, labor, and birth has not been fully observed [16]. Several studies have obtained an overview of the estrogenic effect of phytoestrogens [17].…”

Section: Introductionmentioning

confidence: 99%

The Effect of Anastatica hierochuntica L. Extract on the Histology of Myometrial Cells and Prostaglandin levels (PGE2, PGF2α) in Pregnant Mice

Astutik¹,

Santoso²,

Agil³

2020

Proceedings of the 4th International Symposium on Health Research (ISHR 2019)

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Anastatica hierochuntica L. is well known as a traditional medicine which brings benefit to the final trimester of pregnancy by increase uterine contractions. This study aimed to analyze the effect of A. hierochuntica L. extract on the histology of myometrial cells and prostaglandin levels (PGE2 and PGF2α) of pregnant mice. The mice were divided into three groups: (1) control group, treated by 0.5% CMC solution 10 ml/kg bw/day; (2) two treatment groups that treated by A. hierochuntica L. extract with 100 and 150 mg/kg bw/day doses, respectively. Uterine tissue was collected for histological analysis and blood serum was collected to examine the PGE2 and PGF2α levels. It obtained that myometrial hypertrophy increased significantly in the treatment groups (p = 0.001) than in the control group. Moreover, there were significant differences in the PGE2 levels (p = 0.001) and PGF2α levels (p = 0.000) between the control group and the treatment groups. The 100 mg/kg bw/day dose had a greater effect on myometrial hypertrophy than 150 mg/kg bw/day, which resulted in a higher increase of PGE2 and PGF2α levels. Hence, it was concluded that the 100 mg/kg bw/day dose of A. hierochuntica L. extract was the appropriate dose for increasing prostaglandin levels.

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Methods and Model Systems Used to Study Pregnant Human Uterine Smooth Muscle

Cited by 4 publications

References 89 publications

Identification of mundulone and mundulone acetate as natural products with tocolytic efficacy in mono- and combination-therapy with current tocolytics

Identification of mundulone and mundulone acetate as natural products with tocolytic efficacy in mono- and combination-therapy with current tocolytics

Tools, techniques, and future opportunities for characterizing the mechanobiology of uterine myometrium

The Effect of Anastatica hierochuntica L. Extract on the Histology of Myometrial Cells and Prostaglandin levels (PGE2, PGF2α) in Pregnant Mice

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