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Peer-reviewedCC BY 4.0Nutrients

Whey Protein–Polysaccharide Nanocomplexes: Structure–Function Characterization and Bone-Related Outcomes in Ovariectomized Rats

8 affiliations
  1. 1Laboratorio de Investigación Química y Farmacológica de Productos Naturales, Facultad de Química, Universidad Autónoma de Querétaro, Cerro de las Campanas S/N, Santiago de Querétaro C.P. 76017, Querétaro, Mexico
  2. 2Laboratorio de Procesos en Ingeniería Agroalimentaria, Unidad de Investigación Multidisciplinaria (UIM), FES-Cuautitlán, Universidad Nacional Autónoma de México, Cuautitlán Izcalli C.P. 54714, State of Mexico, Mexico
  3. 3Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Campus Juriquilla, Santiago de Querétaro C.P. 76230, Querétaro, Mexico
  4. 4Escuela de Bachilleres, Plantel Norte, Universidad Autónoma de Querétaro, Av Cerro Sombrerete, Desarrollo San Pablo S/N, Santiago de Querétaro C.P. 76125, Querétaro, Mexico
  5. 5Departamento de Ingeniería Física, División de Ciencias e Ingenierías, Universidad de Guanajuato, Campus León, Lomas del Bosque 103, Col. Lomas del Campestre, León C.P. 37150, Guanajuato, Mexico
  6. 6Departamento de Ingeniería Electromecánica, Tecnológico Nacional de México/ITS de Purísima del Rincón, Blvd. Del Valle 2301, Col. Guardarrayas, Purísima del Rincón C.P. 36413, Guanajuato, Mexico
  7. 7División de Ciencias Biológicas y de La Salud, Departamento de Sistemas Biológicos, Unidad Xochimilco, Universidad Autónoma Metropolitana, Mexico City C.P. 04960, Mexico
  8. 8Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Campus Juriquilla, Santiago de Querétaro C.P. 76230, Querétaro, Mexico

Abstract

Background/Objectives: Estrogen deficiency is associated with impaired bone mineralization and deterioration of bone quality. Protein–polysaccharide nanostructures have attracted interest as food-grade formulations because their physicochemical properties can be modified by the nature and organization of their constituent biopolymers. This study evaluated the effects of whey protein–polysaccharide nanocomplex formulations on bone-related outcomes in ovariectomized female Wistar rats. Methods: Whey protein nanocomplexes containing either Opuntia ficus-indica mucilage or Tamarindus indica xyloglucan were produced by complex coacervation and ultrasonication. The nanocomplexes were characterized in terms of particle size, polydispersity index, ζ-potential, and morphology. Ovariectomized rats received the nanostructured formulations, their corresponding physical mixtures, the individual ingredients, or control diets for 12 weeks. Femoral bone quality was evaluated through biomechanical testing, calcium and phosphorus content, Ca/P ratio, bone mineral density, scanning electron microscopy and histomorphometry, and X-ray diffraction analysis. Results: The developed nanocomplexes exhibited particle sizes below 500 nm and polydispersity indices of approximately 0.3. Their physicochemical characteristics and morphology differed according to the polysaccharide incorporated. Ovariectomy was associated with unfavorable changes in bone mineralization, microarchitecture, crystallographic characteristics, and biomechanical competence. Among the evaluated treatments, the whey protein–O. ficus-indica mucilage nanostructured formulation (LMCC) showed the most favorable overall pattern across most bone-related outcomes, including bone mineral density, femoral calcium content, microarchitecture, hydroxyapatite crystallite size, and biomechanical properties. However, the response to nanostructuring was not uniform across the two polysaccharide systems, and the xyloglucan-based formulations exhibited parameter-dependent responses. Conclusions: The biological response to whey protein–polysaccharide formulations was associated with polysaccharide type and physicochemical organization. The mucilage-based nanostructured formulation showed a more favorable overall pattern across most evaluated bone-related outcomes in ovariectomized rats. However, calcium association within the nanostructures, calcium-release kinetics, gastrointestinal bioaccessibility, and intestinal calcium absorption were not evaluated; therefore, the mechanisms underlying these responses remain to be established. These findings should be interpreted as preclinical and should not be extrapolated directly to osteoporosis prevention in humans.

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BibTeX
@article{rojasmolina2026whey,
  title = {{Whey Protein–Polysaccharide Nanocomplexes: Structure–Function Characterization and Bone-Related Outcomes in Ovariectomized Rats}},
  author = {Juana Isela Rojas-Molina and Elsa Gutiérrez-Cortez and Ricardo Moisés González-Reza and Omar M. Gomez-Vazquez and Brandon Correa-Piña and Ezequiel Hernández-Becerra and Daniel Jiménez-Mendoza and Alejandra Rojas-Molina and Carlos Quirino-Barreda and Michelle Quintero-García},
  journal = {Nutrients},
  year = {2026},
  month = oct,
  doi = {10.3390/nu18203321},
  url = {https://www.mdpi.com/2072-6643/18/20/3321},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Whey Protein–Polysaccharide Nanocomplexes: Structure–Function Characterization and Bone-Related Outcomes in Ovariectomized Rats
AU  - Juana Isela Rojas-Molina
AU  - Elsa Gutiérrez-Cortez
AU  - Ricardo Moisés González-Reza
AU  - Omar M. Gomez-Vazquez
AU  - Brandon Correa-Piña
AU  - Ezequiel Hernández-Becerra
AU  - Daniel Jiménez-Mendoza
AU  - Alejandra Rojas-Molina
AU  - Carlos Quirino-Barreda
AU  - Michelle Quintero-García
PY  - 2026
DA  - 2026/10/01
JO  - Nutrients
DO  - 10.3390/nu18203321
UR  - https://www.mdpi.com/2072-6643/18/20/3321
AB  - Background/Objectives: Estrogen deficiency is associated with impaired bone mineralization and deterioration of bone quality. Protein–polysaccharide nanostructures have attracted interest as food-grade formulations because their physicochemical properties can be modified by the nature and organization of their constituent biopolymers. This study evaluated the effects of whey protein–polysaccharide nanocomplex formulations on bone-related outcomes in ovariectomized female Wistar rats. Methods: Whey protein nanocomplexes containing either Opuntia ficus-indica mucilage or Tamarindus indica xyloglucan were produced by complex coacervation and ultrasonication. The nanocomplexes were characterized in terms of particle size, polydispersity index, ζ-potential, and morphology. Ovariectomized rats received the nanostructured formulations, their corresponding physical mixtures, the individual ingredients, or control diets for 12 weeks. Femoral bone quality was evaluated through biomechanical testing, calcium and phosphorus content, Ca/P ratio, bone mineral density, scanning electron microscopy and histomorphometry, and X-ray diffraction analysis. Results: The developed nanocomplexes exhibited particle sizes below 500 nm and polydispersity indices of approximately 0.3. Their physicochemical characteristics and morphology differed according to the polysaccharide incorporated. Ovariectomy was associated with unfavorable changes in bone mineralization, microarchitecture, crystallographic characteristics, and biomechanical competence. Among the evaluated treatments, the whey protein–O. ficus-indica mucilage nanostructured formulation (LMCC) showed the most favorable overall pattern across most bone-related outcomes, including bone mineral density, femoral calcium content, microarchitecture, hydroxyapatite crystallite size, and biomechanical properties. However, the response to nanostructuring was not uniform across the two polysaccharide systems, and the xyloglucan-based formulations exhibited parameter-dependent responses. Conclusions: The biological response to whey protein–polysaccharide formulations was associated with polysaccharide type and physicochemical organization. The mucilage-based nanostructured formulation showed a more favorable overall pattern across most evaluated bone-related outcomes in ovariectomized rats. However, calcium association within the nanostructures, calcium-release kinetics, gastrointestinal bioaccessibility, and intestinal calcium absorption were not evaluated; therefore, the mechanisms underlying these responses remain to be established. These findings should be interpreted as preclinical and should not be extrapolated directly to osteoporosis prevention in humans.
SN  - 2072-6643
ER  -