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Vitamin D nanoemulsion corrected deficiency and improved bones in 1 week (high dose in rats) – Jan 2019

Stabilization of Vitamin D in Pea Protein Isolate Nanoemulsions Increases Its Bioefficacy in Rats

Nutrients 2019, 11(1), 75; https://doi.org/10.3390/nu11010075 (registering DOI)
Ali M. Almajwal 1,* , Mahmoud M. A. Abulmeaty 1,2, Hao Feng 3, Nawaf W. Alruwaili 3, Astrid Dominguez-Uscanga 3, Juan E. Andrade 3,*, Suhail Razak 1 and Mohamed F. ElSadek 1
1 Dept of Community Health Sciences, College of Applied Medical Sciences, King Saud University, Riyadh 11433, Saudi Arabia
2 Dept of Medical Physiology, Faculty of Medicine, Zagazig University, Zagazig 44519, Egypt
3 Dept of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Champaign, IL 61801, USA
(This article belongs to the Special Issue Food Fortification: Traditional and New Strategies to Combat Micronutrient Deficiencies)

  • This is one of many studies on non-emulsion of vitamin D, etc.
  • They started with Vitamin D deficient rats
  • They gave a high dose every 2nd day for just one week
  • Huge increase in levels in blood vs virtually no increase for rats getting just Vitamin D
  • Also, big decrease in bone problems vs controls, again in just one week

Clipped from PDF

  • "The increase in serum 25(OH)VitD was 2.3 times higher than its control (Nano-control) after one-week of treatment, potentially due to increased protection of the vitamin during digestion and enhancement of oral bioavailability due to improved micellarization of VitD in the small intestine "
  • "The group receiving Oil-VitD did not improve 25(OH)VitD status. This could be due to lower absorption"
  • "Oral supplementation of VitD dispersed in nanoparticles in water increases circulating levels within three days "

See also VitaminDWiki

 Download the PDF from VitaminDWiki
Table 2. Concentration of several blood biomarkers of VitD deficiency in rats after one week of dietary treatments.

GROUPWhat it means
Typically for week 6
ControlRats fed Vitamin D
from birth
Nano-control Nano, without Vit D
Oil-controlOil, without Vit D
Nano-VitDNano + Vitamin D
Oil-VitDOil + Vitamin D

Daily nanoemulsion improved bone measureents in just 1 week!!

Table 3. Histomorphometric parameters among study groups.
.Note: Improved bone in growing rats does not imply improved bone in elderly humans (osteoporosis)

Micronutrient delivery formulations based on nanoemulsions can enhance the absorption of nutrients and bioactives, and thus, are of great potential for food fortification and supplementation strategies. The aim was to evaluate the bioefficacy of vitamin D (VitD) encapsulated in nanoemulsions developed by sonication and pH-shifting of pea protein isolate (PPI) in restoring VitD status in VitD-deficient rats. Weaned male albino rats (n = 35) were fed either normal diet AIN-93G (VitD 1000 IU/kg) (control group; n = 7) or a VitD-deficient diet (<50 IU/kg) for six weeks (VitD-deficient group; n = 28). VitD-deficient rats were divided into four subgroups (n = 7/group). Nano-VitD and Oil-VitD groups received a dose of VitD (81 µg) dispersed in either PPI-nanoemulsions or in canola oil, respectively, every other day for one week. Their control groups, Nano-control and Oil-control, received the respective delivery vehicles without VitD. Serum 25-hydroxyvitamin D [25(OH)VitD], parathyroid hormone (PTH), Ca, P, and alkaline phosphatase (ALP) activity were measured. After one week of treatment, the VitD-deficient rats consuming Nano-VitD recovered from Vitamin D deficiency (VDD) as compared against baseline and had serum 25(OH)VitD higher than the Nano-control. Enhancement in VitD status was followed with expected changes in serum PTH, Ca, P, and ALP levels, as compared against the controls. Stabilization of VitD within PPI-based nanoemulsions enhances its absorption and restores its status and biomarkers of bone resorption in VitD-deficient rats.


  1. Roth, D.E.; Abrams, S.A.; Aloia, J.; Bergeron, G.; Bourassa, M.W.; Brown, K.H.; Calvo, M.S.; Cashman, K.D.; Combs, G.; De-Regil, L.M.; et al. Global prevalence and disease burden of vitamin D deficiency: A roadmap for action in low- and middle-income countries. Ann. N. Y. Acad. Sci. 2018,1430,44-79.
  2. Mithal, A.; Wahl, D.A.; Bonjour, J.P.; Burckhardt, P.; Dawson-Hughes, B.; Eisman, J.A.; El-Hajj Fuleihan, G.; Josse, R.G.; Lips, P.; Morales-Torres, J. Global vitamin D status and determinants of hypovitaminosis D. Osteoporos. Int. 2009,20,1807-1820.
  3. Holick, M.F. High prevalence of vitamin D inadequacy and implications for health. Mayo Clin. Proc. 2006, 81, 353-373.
  4. Wang, H.; Chen, W.; Li, D.; Yin, X.; Zhang, X.; Olsen, N.; Zheng, S.G. Vitamin D and Chronic Diseases. Aging Dis. 2017, 8, 346-353.
  5. Haines, S.T.; Park, S.K. Vitamin D supplementation: What's known, what to do, and what's needed. Pharmacotherapy 2012, 32, 354-382.
  6. Cashman, K.D.; Kiely, M. Vitamin D and Food Fortification. In Vitamin D: Health, Disease and Therapeutics, 4th ed.; Academic Press: London, UK, 2018; Volume 2, pp. 109-127. ISBN 9780128099636.
  7. Park, S.J.; Garcia, C.V.; Shin, G.H.; Kim, J.T. Development of nanostructured lipid carriers for the encapsulation and controlled release of vitamin D3. Food Chem. 2017, 225, 213-219.
  8. Shu, G.; Khalid, N.; Zhao, Y.; Neves, M.A.; Kobayashi, I.; Nakajima, M. Formulation and stability assessment of ergocalciferol loaded oil-in-water nanoemulsions: Insights of emulsifiers effect on stabilization mechanism. Food Res. Int. 2016, 90, 320-327.
  9. Ozturk, B.; Argin, S.; Ozilgen, M.; McClements, D.J. nanoemulsion delivery systems for oil-soluble vitamins: Influence of carrier oil type on lipid digestion and vitamin D3 bioaccessibility. Food Chem. 2015,187, 499-506.
  10. Mohammadi, M.; Ghanbarzadeh, B.; Hamishehkar, H. Formulation of nanoliposomal vitamin D3 for potential application in beverage fortification. Adv. Pharm. Bull. 2014, 4, 569-575.
  11. Diarrassouba, F.; Garrait, G.; Remondetto, G.; Alvarez, P.; Beyssac, E.; Subirade, M. Food protein-based microspheres for increased uptake of Vitamin D3. Food Chem. 2015,173,1066-1072.
  12. Hasanvand, E.; Fathi, M.; Bassiri, A. Production and characterization of vitamin D3-loaded starch nanoparticles: Effect of amylose to amylopectin ratio and sonication parameters. J. Food Sci. Technol. 2018, 55,1314-1324.
  13. Teng, Z.; Luo, Y.; Wang, Q. Carboxymethyl chitosan-soy protein complex nanoparticles for the encapsulation and controlled release of vitamin D3. Food Chem. 2013,141, 524-532.
  14. Sharma, A.; Sharma, U.S. Liposomes in drug delivery: Progress and limitations. Int. J. Pharm. 1997,154, 123-140.
  15. McClements, D.J. Encapsulation, protection, and release of hydrophilic active components: Potential and limitations of colloidal delivery systems. Adv. Colloid Interface Sci. 2015, 219, 27-53.
  16. Jain, K.; Kumar Mehra, N.; Jain, N.K. nanotechnology in drug delivery: Safety and toxicity issues. Curr. Pharm. Des. 2015, 21, 4252-4261.
  17. Wang, Q.; Allen, J.C.; Swaisgood, H.E. Binding of Vitamin D and Cholesterol to |3-Lactoglobulin. J. Dairy Sci. 1997, 80,1054-1059.
  18. Yang, M.C.; Guan, H.H.; Liu, M.Y.; Lin, Y.H.; Yang, J.M.; Chen, W.L.; Chen, C.J.; Mao, S.J.T. Crystal structure of a secondary vitamin D3-binding site of milk |3-lactoglobulin. Proteins Struct. Funct. Genet. 2008, 71, 1197-1210.
  19. Yang, M.C.; Guan, H.H.; Yang, J.M.; Ko, C.N.; Liu, M.Y.; Lin, Y.H.; Huang, Y.C.; Chen, C.J.; Mao, S.J.T. Rational design for crystallization of |3-lactoglobulin and vitamin D3 complex: Revealing a secondary binding site. Cryst. Growth Des. 2008, 8, 4268-4276.
  20. Kohl, E.A.; Schaefer, P.C. Improved high-pressure liquid chromatographic assay of serum 25-hydroxycholecalciferol and 25-hydroxyergocalctferol after reverse-phase sep-pak C18cartridge preparation of sample. J. Liq. Chromatogr. 1981, 4, 2023-2037.
  21. Kao, P.C.; Heser, D.W. Simultaneous determination of 25-hydroxy- and 1,25-dihydroxyvitamin D from a single sample by dual-cartridge extraction. Clin. Chem. 1984, 30, 56-61.
  22. Foegeding, E.A.; Davis, J.P. Food protein functionality: A comprehensive approach. Food Hydrocoll. 2011, 25, 1853-1864.
  23. Lam, R.S.H.; Nickerson, M.T. Food proteins: A review on their emulsifying properties using a structure-function approach. Food Chem. 2013,141, 975-984.
  24. Beverung, C.J.; Radke, C.J.; Blanch, H.W. Protein adsorption at the oil/water interface: Characterization of adsorption kinetics by dynamic interfacial tension measurements. Biophys. Chem. 1999, 81, 59-80.
  25. Karaca, A.C.; Low, N.; Nickerson, M. Emulsifying properties of chickpea, faba bean, lentil and pea proteins produced by isoelectric precipitation and salt extraction. Food Res. Int. 2011, 44, 2742-2750.
  26. Jiang, S.; Ding, J.; Andrade, J.; Rababah, T.M.; Almajwal, A.; Abulmeaty, M.M.; Feng, H. Modifying the physicochemical properties of pea protein by pH-shifting and ultrasound combined treatments. Ultrason. Sonochem. 2017, 38, 835-842.
  27. Lee, H.; Yildiz, G.; dos Santos, L.C.; Jiang, S.; Andrade, J.E.; Engeseth, N.J.; Feng, H. Soy protein nano-aggregates with improved functional properties prepared by sequential pH treatment and ultrasonication. Food Hydrocoll. 2016, 55, 200-209.
  28. Fleet, J.C.; Gliniak, C.; Zhang, Z.; Xue, Y.; Smith, K.B. Serum metabolite profiles and target tissue gene expression define the effect of cholecalciferol intake on calcium metabolism in rats and mice. J. Nutr. 2008, 138,1114-1120.
  29. Reeves, P.G.; Nielsen, F.H.; Fahey, G.C. AIN-93 Purified Diets for Laboratory Rodents: Final Report of the American Institute of Nutrition Ad Hoc Writing Committee on the Reformulation of the AIN-76A Rodent Diet. J. Nutr. 1993,123,1939-1951.
  30. Itakura, C.; Yamasaki, K.; Goto, M. Pathology of experimental vitamin D deficiency rickets in growing chickens. II. Parathyroid gland. Avian Pathol. 1978, 7, 515-532.
  31. Egan, K.P.; Brennan, T.A.; Pignolo, R.J. Bone histomorphometry using free and commonly available software. Histopathology 2012, 61,1168-1173.
  32. Holick, M.F. The vitamin D deficiency pandemic: Approaches for diagnosis, treatment and prevention. Rev. Endocr. Metab. Disord. 2017,18,153-165.
  33. Salvia-Trujillo, L.; Martin-Belloso, O.; McClements, D. Excipient nanoemulsions for Improving Oral Bioavailability of Bioactives. nanomaterials 2016, 6, 17.
  34. Kadappan, A.S.; Guo, C.; Gumus, C.E.; Bessey, A.; Wood, R.J.; McClements, D.J.; Liu, Z. The Efficacy of nanoemulsion-Based Delivery to Improve Vitamin D Absorption: Comparison of In Vitro and In Vivo Studies. Mol. Nutr. Food Res. 2018, 62,1700836.
  35. Lee, Y.-J.; Kwon, M.; Kim, T.-H.; Kim, K.; Jeong, S.-H.; Chang, H.-R. Pharmacokinetic Characterization of nano-emulsion Vitamin A, D and E (LaVita) in Rats. Korean J. Environ. Agric. 2011, 30,196-201.
  36. Diarrassouba, F.; Garrait, G.; Remondetto, G.; Alvarez, P.; Beyssac, E.; Subirade, M. Improved bioavailability of vitamin D3 using a |3-lactoglobulin-based coagulum. Food Chem. 2015,172, 361-367.
  37. European Chemical Agency. Regulation (EU) No 528/2012 Concerning the Making Available on the Market and Use of biocidal Products. Assessment Report: Cholecalciferol, PT 14 (Rodenticides). 2018. Available online: https://echa.europa.eu/documents/10162/652777b2-d738-a2e8-7b1d-431fd61ba82f (accessed on 28 December 2018).
  38. Toromanoff, A.; Ammann, P.; Mosekilde, L.; Thomsen, J.S.; Riond, J.L. Parathyroid hormone increases bone formation and improves mineral balance in vitamin D-deficient female rats. Endocrinology 1997,138, 2449-2457.
  39. Khundmiri, S.J.; Murray, R.D.; Lederer, E. PTH and Vitamin D. Compr. Physiol. 2016, 6, 561-601.
  40. Lips, P.; Van Schoor, N.M. The effect of vitamin D on bone and osteoporosis. Best Pract. Res. Clin. Endocrinol. Metab. 2011, 25,585-591.
  41. Millan, J.L. The role of phosphatases in the initiation of skeletal mineralization. Calcif. Tissue Int. 2013, 93, 299-306.
  42. Chuang, L.H.; Tung, Y.C.; Liu, S.Y.; Lee, C.T.; Chen, H.L.; Tsai, W.Y. Nutritional rickets in Taiwanese children: Experiences at a single center. J. Formos. Med. Assoc. 2018,117, 583-587.
  43. Bhambri, R.; Naik, V.; Malhotra, N.; Taneja, S.; Rastogi, S.; Ravishanker, U.; Mithal, A. Changes in Bone Mineral Density Following Treatment of Osteomalacia. J. Clin. Densitom. 2006, 9, 120-127.
  44. Shaheen, S.; Noor, S.S.; Barakzai, Q. Serum alkaline phosphatase screening for vitamin D deficiency states. J. Coll. Phys. Surg. Pak. 2012, 22, 424-427.
  45. Uchida, H.; Kurata, Y.; Hiratsuka, H.; Umemura, T. The effects of a vitamin D—Deficient diet on chronic cadmium exposure in rats. Toxicol. Pathol. 2010, 38, 730-737.
  46. DeLuca, H.F. Overview of general physiologic features and functions of vitamin D. Am. J. Clin. Nutr. 2004, 80, 1689S-1696S.
  47. Lips, P. Vitamin D physiology. Prog. Biophys. Mol. Biol. 2006, 92, 4-8.
  48. Need, A.G.; O'Loughlin, P.D.; Morris, H.A.; Coates, P.S.; Horowitz, M.; Nordin, B.E.C. Vitamin D metabolites and calcium absorption in severe vitamin D deficiency. J. Bone Miner. Res. 2008,23,1859-1863.
  49. Brautbar, N.; Walling, M.W.; Coburn, J.W. Interactions between vitamin D deficiency and phosphorus depletion in the rat. J. Clin. Investig. 1979, 63, 335-341.
  50. Coburn, J.W.; Massry, S.G. Changes in serum and urinary calcium during phosphate depletion: Studies on mechanisms. J. Clin. Investig. 1970, 49,1073-1087.
  51. Clark, I.; Rivera-Cordero, F. Effects of endogenous parathyroid hormone on calcium, magnesium and phosphate metabolism in rats. II. Alterations in dietary phosphate. Endocrinology 1974, 95, 360-369.
  52. Steele, T.H. Renal resistance to parathyroid hormone during phosphorus deprivation. J. Clin. Investig. 1976, 58, 1461-1464.
  53. Coburn, K.R. Preliminary investigation of bone change as a result of exposure to reduced atmospheric pressure. Aerosp. Med. 1970, 41,188-190.
  54. Rader, J.I.; Howard, G.A.; Feist, E.; Turner, R.T.; Baylink, D.J. Bone mineralization and metabolism of 3H-25-hydroxyvitamin D3 in thyroparathyroidectomized rats treated with parathyroid extract. Calcif. Tissue Int. 1979, 29, 21-26.
  55. Khaw, K.; Sneyd, M.; Compston, J. Bone density, parathyroid hormone and 25-hydroxyvitaminD concentrations in middle-aged women. Br. Med. J. 1992, 305, 273-277.
  56. Bonifacio, B.V.; da Silva, P.B.; Aparecido dos Santos Ramos, M.; Maria Silveira Negri, K.; Maria Bauab, T.; Chorilli, M. nanotechnology-based drug delivery systems and herbal medicines: A review. Int. J. nanomedicine 2013, 9,1-15.
  57. Silva, A.C.; Santos, D.; Ferreira, D.; Lopes, C.M. Lipid-Based nanocarriers as an Alternative for Oral Delivery of Poorly Water-Soluble Drugs: Peroral and Mucosal Routes. Curr. Med. Chem. 2012,4495-4510.
  58. McClements, D.J. Edible lipid nanoparticles: Digestion, absorption, and potential toxicity. Prog. Lipid Res. 2013,52,409-423.
  59. Yen, C.C.; Chen, Y.C.; Wu, M.T.; Wang, C.C.; Wu, Y.T. nanoemulsion as a strategy for improving the oral bioavailability and anti-inflammatory activity of andrographolide. Int. J. nanomedicine 2018,13, 669-680.
  60. Sun, L.; Wan, K.; Hu, X.; Zhang, Y.; Yan, Z.; Feng, J.; Zhang, J. Functional nanoemulsion-hybrid lipid nanocarriers enhance the bioavailability and anti-cancer activity of lipophilic diferuloylmethane. nanotechnology 2016, 27, 085102.
  61. Cheong, A.M.; Tan, C.P.; Nyam, K.L. Effect of Emulsification Method and Particle Size on the Rate of in vivo Oral Bioavailability of Kenaf (Hibiscus cannabinus L.) Seed Oil. J. Food Sci. 2018, 83,1964-1969.
  62. Saratale, R.G.; Lee, H.-S.; Koo, Y.E.; Saratale, G.D.; Kim, Y.J.; Imm, J.Y.; Park, Y. Absorption kinetics of vitamin E nanoemulsion and green tea microstructures by intestinal in situ single perfusion technique in rats. Food Res. Int. 2018,106,149-155.
  63. Belhaj, N.; Dupuis, F.; Arab-Tehrany, E.; Denis, F.M.; Paris, C.; Lartaud, I.; Linder, M. Formulation, characterization and pharmacokinetic studies of coenzyme Q10PUFA's nanoemulsions. Eur. J. Pharm. Sci. 2012, 47, 305-312.
  64. Kralova, I.; Sjoblom, J. Surfactants used in food industry: A review. J. Dispers. Sci. Technol. 2009, 30, 1363-1383.
  65. McClements, D.J.; Rao, J. Food-Grade nanoemulsions: Formulation, fabrication, properties, performance, Biological fate, and Potential Toxicity. Crit. Rev. Food Sci. Nutr. 2011, 51, 285-330.
  66. Yerramilli, M.; Longmore, N.; Ghosh, S. Stability and Bioavailability of Curcumin in Mixed Sodium Caseinate and Pea Protein Isolate nanoemulsions. J. Am. Oil Chem. Soc. 2018, 95,1013-1026.
  67. Malekzad, H.; Mirshekari, H.; Sahandi Zangabad, P.; Moosavi Basri, S.M.; Baniasadi, F.; Sharifi Aghdam, M.; Karimi, M.; Hamblin, M.R. Plant protein-based hydrophobic fine and ultrafine carrier particles in drug delivery systems. Crit. Rev. Biotechnol. 2018, 38, 47-67.
  68. Glerup, H. Vitamin D deficiency among immigrants. Ugeskr. Laeger 2000,162, 6196-6199.
  69. Kauppinen-Makelin, R.; Tahtela, R.; Loyttyniemi, E.; Karkkainen, J.; Valimaki, M.J. A high prevalence of hypovitaminosis D in Finnish medical in- and outpatients. J. Intern. Med. 2001, 249, 559-563.
  70. Thomas, M.K.; Lloyd-Jones, D.M.; Thadhani, R.I.; Shaw, A.C.; Deraska, D.J.; Kitch, B.T.; Vamvakas, E.C.; Dick, I.M.; Prince, R.L.; Finkelstein, J.S. Hypovitaminosis D in Medical Inpatients. N. Engl. J. Med. 1998, 338, 777-783.
  71. Marshall, I.; Mehta, R.; Ayers, C.; Dhumal, S.; Petrova, A. Prevalence and risk factors for vitamin D insufficiency and deficiency at birth and associated outcome. BMC Pediatr. 2016,16, 208.
  72. Kift, R.; Berry, J.L.; Vail, A.; Durkin, M.T.; Rhodes, L.E.; Webb, A.R. Lifestyle factors including less cutaneous sun exposure contribute to starkly lower vitamin D levels in U.K. South Asians compared with the white population. Br. J. Dermatol. 2013,169,1272-1278.
  73. Clemens, T.L.; Henderson, S.L.; Adams, J.S.; Holick, M.F. Incerased skin pigment reduces the capacity of skin to synthesise vitamin D3. Lancet 1982, 319, 74-76.
  74. Al Jurayyan, N.A.; Mohamed, S.; Al Issa, S.D.; Al Jurayyan, A.N. Rickets and osteomalacia in Saudi children and adolescents attending endocrine clinic, Riyadh, Saudi Arabia. Sudan. J. Paediatr. 2012,12,56-63.
  75. Gonnet, M.; Lethuaut, L.; Boury, F. New trends in encapsulation of liposoluble vitamins. J. Control. Release 2010,146, 276-290.
  76. Holick, M.F. Calcium and vitamin D. Diagnostics and therapeutics. Clin. Lab. Med. 2000,20,569-590.
  77. Murphy, S.C.; Whited, L.J.; Rosenberry, L.C.; Hammond, B.H.; Bandler, D.K.; Boor, K.J. Fluid milk vitamin fortification compliance in New York State. J. Dairy Sci. 2001, 84, 2813-2820.
  78. Marshall, E.F. Cholecalciferol: A unique toxicant for rodent control. In Proceedings of the Eleventh Vertebrate Pest Conference, Sacramento, CA, USA, 6-8 March 1984.
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