Causes of Low Milk Supply: The Roles of Estrogens, Progesterone, and Related External Factors.

Low milk supply (LMS) poses a significant challenge to exclusive and continued breastfeeding, affecting ∼10% to 15% of mothers. Milk production is intricately regulated by both endocrine and autocrine control mechanisms, with estrogens and progesterone playing pivotal roles in this process. In addition to endogenously produced hormones, external substances capable of interfering with normal hormonal actions, including phytoestrogens, mycoestrogens, synthetic estrogens, and hormonal contraceptives, can influence milk production. The effects of these extrinsic hormones on milk production may vary based on maternal body mass index. This comprehensive review examines the multifaceted causes of LMS, focusing on the involvement of estrogens, progesterone, and related external factors in milk production. Furthermore, it investigates the interplay between hormonal factors and obesity, aiming to elucidate the endocrine mechanisms underlying obesity-associated LMS. Insights from this review provide valuable perspectives for developing interventions to improve milk production and address the challenges associated with LMS.

Oestrogens and progesterone in human milk and their effects on infant health outcomes: A narrative review.

Human milk (HM) is a complex biological system that contains a wide range of bioactive components including oestrogens and progesterone. Whilst maternal oestrogens and progesterone concentrations drop rapidly after birth, they remain detectable in HM across lactation. Phytoestrogens and mycoestrogens, which are produced by plants and fungi, are also present in HM and can interact with oestrogen receptors to interfere with normal hormone functions. Despite the potential impact of HM oestrogens and progesterone on the infant, limited research has addressed their impact on the growth and health of breastfed infants. Furthermore, it is important to comprehensively understand the factors that contribute to these hormone levels in HM, in order to establish effective intervention strategies. In this review, we have summarized the concentrations of naturally occurring oestrogens and progesterone in HM from both endogenous and exogenous sources and discussed both maternal factors impacting HM levels and relationships with infant growth.

Human Milk Metabolic Hormones: Analytical Methods and Current Understanding.

Human milk (HM) contains a wide array of peptide hormones including leptin and adiponectin, which are involved in the regulation of infant growth and development. These essential hormones might play an important role in the regulation of metabolic reprogramming of the new-born infant. However, HM hormone studies are sparse and heterogeneous in regard to the study design, sample collection, preparation and analysis methods. This review discussed the limitations of HM hormone analysis highlighting the gaps in pre-analytical and analytical stages. The methods used to quantify HM metabolic hormones (leptin, adiponectin, ghrelin, insulin, obestatin, resistin and apelin) can be classified as immunoassay, immunosensor and chromatography. Immunoassay methods (ELISA and RIA) have been predominantly used in the measurement of these HM hormones. The relative validity parameters of HM hormones analysis are often overlooked in publications, despite the complexity and differences of HM matrix when compared to that of plasma and urine. Therefore, appropriate reports of validation parameters of methodology and instrumentation are crucial for accurate measurements and therefore better understanding of the HM metabolic hormones and their influences on infant outcomes.

Effect of nipple shield use on milk removal: a mechanistic study.

BACKGROUND: Concerns about reduced milk transfer with nipple shield (NS) use are based on evidence from studies with methodological flaws. Milk removal during breastfeeding can be impacted by infant and maternal factors other than NS use. The aim of this study was to control electric breast pump vacuum strength, pattern and duration across multiple study sessions to determine if NS use reduces milk removal from the breast. METHODS: A within-subject study with two groups of breastfeeding mothers (infants < 6 months) were recruited; Control Group (CG): no breastfeeding difficulties; Pain Group (PG) used NS for persistent nipple pain. Mothers completed three randomised 15 min pumping sessions using the Symphony vacuum curve (Medela AG); no NS, fitted NS, and a small NS. Sessions were considered valid where the applied vacuum was within 20 mmHg of the set vacuum. Milk removal was considered as pumped milk volume, and also percentage of available milk removed (PAMR), which is calculated as the pumped volume divided by the estimated milk volume stored in the breast immediately prior to pumping. RESULTS: Of 62 sessions (all: n = 31 paired sessions) a total of 11 paired sessions from both PG (n = 03) and CG (n = 08) were valid (subset) with and without a fitted NS. Only 2 small shield sessions were valid and so all small shield measurements were excluded. Both pumped volumes and PAMR were significantly lower with NS use for all data but not for subset data. (All: Volume and PAMR median: no NS: 76.5 mL, 69%, Fitted NS: 32.1 mL, 41% respectively (volume p = 0.002, PAMR p = 0.002); Subset: Volume and PAMR median: no NS: 83.8 mL, 72%; Fitted NS: 35.2 mL, 40% (volume p = 0.111 and PAMR p = 0.045). The difference in PAMR, but not volume, was statistically significant when analysed by linear mixed modelling. A decrease of 10 mmHg was associated with a 4.4% increase in PAMR (p = 0.017). CONCLUSIONS: This experimental data suggests that nipple shield use may reduce milk removal. Close clinical monitoring of breastfeeding mothers using nipple shields is warranted.

Detection of Milk Ejection Using Bioimpedance Spectroscopy in Lactating Women during Milk Expression Using an Electric Breast Pump.

Milk ejection is essential for effective milk removal during breastfeeding and pumping, and for continued milk synthesis. Many women are unable to accurately sense milk ejection to determine whether their infant is receiving milk or, when pumping, to switch the pump to a more effective expression pattern. To determine if changes in bioimpedance parameters are associated with milk ejection in the lactating breast during pumping. 30 lactating women participated in 2 pumping sessions within 2 weeks of each other. During pumping the breasts were monitored with bioimpedance spectroscopy (on either the pumped or the non- pumped breast), and milk flow rate and volume were measured simultaneously. All mothers completed 24-h milk productions. Linear mixed effects models were used to determine associations between milk flow rate and bioimpedance changes. Changes in bioimpedance parameters were greater at the first milk ejection when measured on the pumped breast (median (IQR): R zero: -7 (-17, -4,) % (n = 30); R infinity: -8 (-20, -2) % (n = 29); membrane capacitance: -24 (-59, -7) % (n = 27). Changes in bioimpedance detected in the non-pumped breast were lower at the first milk ejection, R zero: -3 (-8, -2) % (n = 25); R infinity: -5 (-8, -2) % (n = 23); membrane capacitance: -9 (-17, 15) % (n = 24). Smaller less consistent decreases in the bioimpedance characteristics were detected at the second milk ejection in both breasts. Bioimpedance parameters showed a consistent decrease associated with the first milk ejection when electrodes were placed on the pumped breast. Smaller decreases were observed when the non-pumped breast was monitored for the first and second milk ejection. There was wide variation in the magnitude of changes observed, and hence further development of the methodology is needed to ensure reliability.

Human milk miRNAs primarily originate from the mammary gland resulting in unique miRNA profiles of fractionated milk.

Human milk (HM) contains regulatory biomolecules including miRNAs, the origin and functional significance of which are still undetermined. We used TaqMan OpenArrays to profile 681 mature miRNAs in HM cells and fat, and compared them with maternal peripheral blood mononuclear cells (PBMCs) and plasma, and bovine and soy infant formulae. HM cells and PBMCs (292 and 345 miRNAs, respectively) had higher miRNA content than HM fat and plasma (242 and 219 miRNAs, respectively) (p < 0.05). A strong association in miRNA profiles was found between HM cells and fat, whilst PBMCs and plasma were distinctly different to HM, displaying marked inter-individual variation. Considering the dominance of epithelial cells in mature milk of healthy women, these results suggest that HM miRNAs primarily originate from the mammary epithelium, whilst the maternal circulation may have a smaller contribution. Our findings demonstrate that unlike infant formulae, which contained very few human miRNA, HM is a rich source of lactation-specific miRNA, which could be used as biomarkers of the performance and health status of the lactating mammary gland. Given the recently identified stability, uptake and functionality of food- and milk-derived miRNA in vivo, HM miRNA are likely to contribute to infant protection and development.

Levels of innate immune factors in preterm and term mothers' breast milk during the 1st month postpartum.

There is a paucity of data on the effect of preterm birth on the immunological composition of breast milk throughout the different stages of lactation. We aimed to characterise the effects of preterm birth on the levels of immune factors in milk during the 1st month postpartum, to determine whether preterm milk is deficient in antimicrobial factors. Colostrum (days 2-5 postpartum), transitional milk (days 8-12) and mature milk (days 26-30) were collected from mothers of extremely preterm (<28 weeks of gestation, n 15), very preterm (28-<32 weeks of gestation, n 15), moderately preterm (32-<37 weeks of gestation, n 15) and term infants (37-41 weeks of gestation, n 15). Total protein, lactoferrin, secretory IgA, soluble CD14 receptor (sCD14), transforming growth factor-ß2 (TGF-ß2), α defensin 5 (HD5), ß defensins 1 (HBD1) and 2, IL-6, IL-10, IL-13, interferon-γ, TNF-α and lysozyme (LZ) were quantified in milk. We examined the effects of lactation stage, gestational age, volume of milk expressed, mode of delivery, parity and maternal infection on milk immune factor concentrations using repeated-measures regression analysis. The concentrations of all factors except LZ and HD5 decreased over the 1st month postpartum. Extremely preterm mothers had significantly higher concentrations of HBD1 and TGF-ß2 in colostrum than term mothers did. After controlling for other variables in regression analyses, preterm birth was associated with higher concentrations of HBD1, LZ and sCD14 in milk samples. In conclusion, preterm breast milk contains significantly higher concentrations of some immune proteins than term breast milk.

Gene expression in breastmilk cells is associated with maternal and infant characteristics.

Breastmilk is a rich source of cells with a heterogeneous composition comprising early-stage stem cells, progenitors and more differentiated cells. The gene expression profiles of these cells and their associations with characteristics of the breastfeeding mother and infant are poorly understood. This study investigated factors associated with the cellular dynamics of breastmilk and explored variations amongst women. Genes representing different breastmilk cell populations including mammary epithelial and myoepithelial cells, progenitors, and multi-lineage stem cells showed great variation in expression. Stem cell markers ESRRB and CK5, myoepithelial marker CK14, and lactocyte marker α-lactalbumin were amongst the genes most highly expressed across all samples tested. Genes exerting similar functions, such as either stem cell regulation or milk production, were found to be closely associated. Infant gestational age at delivery and changes in maternal bra cup size between pre-pregnancy and postpartum lactation were associated with expression of genes controlling stemness as well as milk synthesis. Additional correlations were found between genes and dyad characteristics, which may explain abnormalities related to low breastmilk supply or preterm birth. Our findings highlight the heterogeneity of breastmilk cell content and its changes associated with characteristics of the breastfeeding dyad that may reflect changing infant needs.

Milk ejection patterns: an intra- individual comparison of breastfeeding and pumping.

BACKGROUND: Milk ejection is a transient episode critical to milk removal and women typically have multiple milk ejections during breastfeeding and pumping. Recently it was found that milk ejection characteristics such as number of milk ejections and periodicity were consistent throughout 12 months of lactation in women who expressed their milk with an electric breast pump. It is not known whether the stimulation of an infant at the breast influences milk ejection patterns or whether this is a programmed event. The aim of this study was to compare milk ejection patterns during breastfeeding and expressing milk with an electric pump within mothers. METHODS: Twelve lactating mothers with normal milk production (502-1356 mL) had milk ejection recorded by measuring the diameter of a major milk duct with ultrasound imaging throughout an entire breastfeed and a 15-min pumping session. Scans were analysed for timing, duration of duct dilation and maximum duct diameter. RESULTS: The initial milk ejection defined as the first increase in duct diameter was observed earlier during breastfeeding than during two phase pumping sessions but was not statistically significant (p = .057). There were no significant differences between the duration of the first or second milk ejection for mothers when breastfeeding or pumping at their maximum comfortable vacuum (p = .18; p = .99). The times taken to reach the peak duct diameter, or the first half of the milk ejection were also not found to be significantly different between breastfeeding and pumping. CONCLUSION: This study suggests that milk ejection patterns remain consistent within individual mothers regardless of whether the mother is breastfeeding or expressing milk indicating a likelihood of the process either being programmed or innate to the individual.

Breastmilk is a novel source of stem cells with multilineage differentiation potential.

The mammary gland undergoes significant remodeling during pregnancy and lactation, which is fuelled by controlled mammary stem cell (MaSC) proliferation. The scarcity of human lactating breast tissue specimens and the low numbers and quiescent state of MaSCs in the resting breast have hindered understanding of both normal MaSC dynamics and the molecular determinants that drive their aberrant self-renewal in breast cancer. Here, we demonstrate that human breastmilk contains stem cells (hBSCs) with multilineage properties. Breastmilk cells from different donors displayed variable expression of pluripotency genes normally found in human embryonic stem cells (hESCs). These genes included the transcription factors (TFs) OCT4, SOX2, NANOG, known to constitute the core self-renewal circuitry of hESCs. When cultured in the presence of mouse embryonic feeder fibroblasts, a population of hBSCs exhibited an encapsulated ESC-like colony morphology and phenotype and could be passaged in secondary and tertiary clonogenic cultures. While self-renewal TFs were found silenced in the normal resting epithelium, they were dramatically upregulated in breastmilk cells cultured in 3D spheroid conditions. Furthermore, hBSCs differentiated in vitro into cell lineages from all three germ layers. These findings provide evidence that breastmilk represents a novel and noninvasive source of patient-specific stem cells with multilineage potential and establish a method for expansion of these cells in culture. They also highlight the potential of these cells to be used as novel models to understand adult stem cell plasticity and breast cancer, with potential use in bioengineering and tissue regeneration.

Blood flow characteristics of the human lactating breast.

BACKGROUND: Despite the increased metabolic activity of the lactating breast, no studies have been carried out to determine mammary blood flow (MBF) parameters or the relationship between MBF and milk production in women. The aim of this study was to measure the MBF in the internal mammary artery (IMA) and lateral thoracic artery (LTA) of lactating women and determine if these were related to milk production. METHODS: Blood flow in the IMA and LTA was measured with color Doppler ultrasound in 55 lactating women. Twenty-four-hour milk production was determined with the test-weigh method. RESULTS: IMA contributed the greater proportion of blood flow to the lactating breast (70%). MBF was highly variable between women but consistent between the left (126 L/24 h; interquartile range, 76-169) and right (110 L/24 h) breasts. No relationship between MBF and milk production was demonstrated. For 3 women, MBF was markedly reduced in 1 breast that was synthesizing almost no milk compared to the other that was producing a normal volume of milk. DISCUSSION: Although no relationship between MBF and milk production was found, the substantial reduction in blood flow in the breasts of lactating women producing almost no milk suggests a threshold below which milk production is compromised. CONCLUSION: Doppler ultrasound did not demonstrate a relationship between MBF and milk production in lactating women. Further investigation is required to fully understand the role of blood flow in milk synthesis.

Tongue movement and intra-oral vacuum of term infants during breastfeeding and feeding from an experimental teat that released milk under vacuum only.

BACKGROUND: Recent literature supports the theory that vacuum is integral to the removal of milk from the breast rather than peristaltic compression of the breast. AIM: We aimed to determine if breastfed infants could remove breast milk from an experimental teat designed to release milk only when a vacuum is applied. METHODS: Submental ultrasound images and intra-oral vacuum measurements were recorded simultaneously during both a breastfeed and a feed with the experimental teat. RESULTS: Infants placed the nipple and teat a similar distance from the nipple hard-soft palate junction when the tongue was lowered (4.7 mm vs 5.3 mm). As the tongue lowered the nipple and teat expanded evenly although the nipple expanded more than the teat (3.1mm vs 1.5 mm). Both baseline (-31 mm Hg vs -12 mm Hg) and peak vacuum (-122 mm Hg vs -67 mm Hg) applied to the breast were significantly higher than for the teat. CONCLUSION: Breastfed infants are able to remove milk from a teat using only vacuum with a similar tongue movement to that of breastfeeding. This evidence supports the theory that vacuum is a critical factor in the removal of milk from the breast.

A comparison of corneal cellular responses after 213-nm compared with 193-nm laser photorefractive keratectomy in rabbits.

PURPOSE: Corneal refractive surgery is typically performed using a 193-nm excimer laser. However, a recently developed 213-nm solid-state (5th harmonic) Nd:YAG laser presents some practical and user safety advantages, although the biological impact of using this wavelength remains poorly characterized. Here, we provide in vivo and in vitro comparisons of the corneal cellular outcomes after irradiation with 213 and 193 nm wavelengths. METHODS: New Zealand White rabbits underwent photorefractive keratectomy with -5 diopters and a 6.5-mm optical zone and studied at time points up to 1 year. The development of haze was examined ophthalmologically and by detecting myofibroblasts immunohistochemically. Cell death was quantified using a terminal deoxynucleotidyl transferase-dUTP nick end labeling (TUNEL) assay, and the number of stromal keratocytes undergoing apoptosis estimated histologically. Superoxide dismutase activity was estimated in vitro by enzyme-linked immunosorbent assay in irradiated rabbit corneal keratocytes. RESULTS: Our results demonstrate subtle differences in the cellular outcomes after irradiation with 213- and 193-nm lasers, despite similar degrees of corneal haze developing in both treatment groups. In vivo, the 213-nm laser results in more stable stromal cell numbers, implying a more predictable ablation outcome. In vitro, higher levels of superoxide dismutase in corneal keratocytes irradiated with 213 nm compared with 193 nm wavelengths suggest a better endogenous protection against free radicals induced by laser surgery. CONCLUSIONS: The more favorable cellular responses after irradiation with 213 nm compared with 193 nm wavelengths are consistent with good clinical outcomes previously reported. Ablation with a 213 nm wavelength may result in better wound healing, leading to a more reliable correction of refractive errors.