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Effects of pasteurization on osteopontin concentrations in human breastmilk

Pediatric Research volume 95, pages 641–646 (2024)Cite this article

Abstract

Background

Osteopontin (OPN) is an important breastmilk protein involved in infant intestinal, immunological, and brain development. However, little is known about how common milk pasteurization and storage techniques affect this important bioactive protein.

Methods

Human milk osteopontin concentration was measured in single-donor fresh (n = 1) or frozen (n = 20) breastmilk, pooled Holder-pasteurized donor breastmilk (n = 11), and a shelf-stable (retort pasteurized) breastmilk product (n = 2) by ELISA. Single-donor breastmilk samples were subjected to pasteurization and/or freezing before measuring osteopontin concentrations.

Results

Holder pasteurization of breastmilk resulted in an ∼50% decrease in osteopontin concentration within single-donor samples. Breastmilk from mothers of preterm infants trended toward higher osteopontin concentration than mothers of term infants; however, samples from preterm mothers experienced greater osteopontin degradation upon pasteurization. A commercial breastmilk product that underwent retort pasteurization had lower osteopontin concentration than a Holder-pasteurized pooled breastmilk product. Finally, freezing breastmilk prior to Holder pasteurization resulted in less osteopontin degradation than Holder pasteurization prior to freezing.

Conclusions

Commonly used breastmilk pasteurization and storage techniques, including freezing and Holder pasteurization, decrease the concentration of the bioactive protein osteopontin in human breastmilk. Holder pasteurization reduced osteopontin concentration by an average of 63%, while freezing resulted in an 8–12% decrease.

Impact

  • Pasteurization of human breastmilk significantly decreases the concentration of the bioactive protein osteopontin.

  • Use of both pasteurization and freezing techniques for breastmilk preservation results in greater loss of osteopontin.

  • This study presents for the first time an analysis of osteopontin concentrations in single-donor pasteurized milk samples.

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Fig. 1: Acquisition and treatment of human breastmilk samples.
Fig. 2: Osteopontin concentrations in the indicated breastmilk products.
Fig. 3: Osteopontin concentrations in frozen single-donor breastmilk from mothers of preterm and term infants.
Fig. 4: Osteopontin concentrations in single-donor breastmilk from mothers of preterm and term infants, before and after Holder pasteurization.
Fig. 5: Osteopontin concentrations in fresh and processed breastmilk.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank Josh McCrary (Monroe Carell Jr. Children’s Hospital at Vanderbilt) for providing pooled donor breastmilk and retort-pasteurized breastmilk samples. Cartoons in Figs. 4 and 5 were created with BioRender.com.

Funding

Funding

This material is based upon work supported by the National Science Foundation Graduate Research Fellowship under Grant No. 1937963 (to K.G.M.), as well as NIH grant R01DK111671 (to D.O-V.); Vanderbilt Training in Cellular, Biochemical and Molecular Sciences Training Program, T32GM008554-25 (to K.G.M.); and the Vanderbilt Institute for Clinical and Translational Research (VICTR) grant VR62082.

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Authors and Affiliations

  1. Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN, USA

    Kathleen G. McClanahan & Danyvid Olivares-Villagómez

  2. Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN, USA

    Jeff Reese & Jörn-Hendrik Weitkamp

  3. Center for Mucosal Inflammation and Cancer, Vanderbilt University Medical Center, Nashville, TN, USA

    Danyvid Olivares-Villagómez

  4. Vanderbilt Institute for Infection, Immunology, and Inflammation, Vanderbilt University Medical Center, Nashville, TN, USA

    Danyvid Olivares-Villagómez

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  1. Kathleen G. McClanahan
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  4. Danyvid Olivares-Villagómez
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Contributions

K.G.M., J.R., and J.-H.W., conceived the project idea; J.-H.W. provided samples; K.G.M. performed data acquisition and analysis; K.G.M. and D.O.-V. wrote the manuscript; J.R., J.-H.W., K.G.M., and D.O.-V. revised and approved the final article.

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Correspondence to Danyvid Olivares-Villagómez.

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All samples were acquired from patients who had given informed consent for their samples to be used for research.

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McClanahan, K.G., Reese, J., Weitkamp, JH. et al. Effects of pasteurization on osteopontin concentrations in human breastmilk. Pediatr Res 95, 641–646 (2024). https://doi.org/10.1038/s41390-023-02838-1

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