Potential of Nanonutraceuticals in Increasing Immunity
Nanomaterials 2020,10,2224; doi:10.3390/nano10112224
Josef Jampilek1,2' josef.jampilek at gmail.com and Katarina Kralova
Department of Analytical Chemistry, Faculty of Natural Sciences, Comenius University, Ilkovicova 6, 842 15 Bratislava, Slovakia
Regional Centre of Advanced Technologies and Materials, Faculty of Science, Palacky University, Slechtitelu 27, 783 71 Olomouc, Czech Republic
Institute of Chemistry, Faculty of Natural Sciences, Comenius University, Ilkovicova 6,
842 15 Bratislava, Slovakia; kata.kralova at gmail.com
Nutraceuticals are defined as foods or their extracts that have a demonstrably positive effect on human health. According to the decision of the European Food Safety Authority, this positive effect, the so-called health claim, must be clearly demonstrated best by performed tests. Nutraceuticals include dietary supplements and functional foods. These special foods thus affect human health and can positively affect the immune system and strengthen it even in these turbulent times, when the human population is exposed to the COVID-19 pandemic. Many of these special foods are supplemented with nanoparticles of active substances or processed into nanoformulations. The benefits of nanoparticles in this case include enhanced bioavailability, controlled release, and increased stability. Lipid-based delivery systems and the encapsulation of nutraceuticals are mainly used for the enrichment of food products with these health-promoting compounds.
This contribution summarizes the current state of the research and development of effective nanonutraceuticals influencing the body's immune responses, such as vitamins (C, D, E, B12, folic acid), minerals (Zn, Fe, Se), antioxidants (carotenoids, coenzyme Q10, polyphenols, curcumin), omega-3 fatty acids, and probiotics.
This study does not appear to mention important features of nano forms
1) Can get into the body without going thru the gut - which is 2x to 4 X faster
2) More gets into the body 2X to 10X more
3) Several of the nanonutraceuticals can be in the same liquid
The founder of VitaminDWiki in March 2021 takes emulsion Omega-3, nanoemulsion vitamin D, and combination Vitamin C and B12 nanoemulsion
Immunity category in VitaminDWiki
see alsoVirus category listing has
Overview Influenza and vitamin D
Vitamin D helps both the innate and adaptive immune systems fight COVID-19 – Jan 2022
Vitamin D aids the clearing out of old cells (autophagy) – many studies
600,000 IU of Vitamin D (total) allowed previously weak immune systems to fight off a virus antigen - Nov 2020
Search for treg OR "t-cell" in VitaminDWiki 1440 items as of Jan 2020
141 VitaminDWiki pages contained "infection" in title (June 2021)
Search VitaminDWik for BACTERIA in title 25 items as of Aug 2019
Vitamin D and the Immune System – chapter Aug 2019
7X less risk of influenza if Vitamin D levels higher than 30 ng – Oct 2017
Common cold prevented and treated by Vitamin D, Vitamin C, Zinc, and Echinacea – review April 2018
Vitamin D improves T Cell immunity – RCT Feb 2016
Immune system - great 11-minute animated video - Aug 2021 nothing about Vitamin D
13 titles in VitaminDWiki contained INNATE or ADAPTIVE as of July 2021
Increasing publications on vitamin D and Infection
COVID-19 treated by Vitamin D - studies, reports, videos
As of June 28, 2022, the VitaminDWiki page had: 34 trials, 11 trial results, 36 meta-analyses and reviews, 69 observations, 38 recommendations, 55 associations, 89 speculations, 58 videos, 45 Mortality studies see related: Governments, HealthProblems, Hospitals, Dark Skins, 26 risk factors are ALL associated with low Vit D, Recent Virus pages Fight COVID-19 with 50K Vit D weekly Vaccines Take lots of Vitamin D at first signs of COVID COVID Clinical Trials (06/22) using Vit D: 114, Vit D & Zinc: 26, Calcidiol: 26
Vitamin D and Omega-3 category starts with
- Vitamin D Receptor is associated in over 40 autoimmune studies
- Vitamin D Receptor activation can be increased by any of: Resveratrol, Omega-3, Magnesium, Zinc, Quercetin, non-daily Vit D, Curcumin, intense exercise, Ginger, Essential oils, etc Note: The founder of VitaminDWiki uses 10 of the 12 known VDR activators
- Vitamin B12 – great improvements with nano forms – March 2021
- Inhaled nanoemulsion of Vitamin D killed lung bacteria – Sept 2017
- Inhaling Vitamin D nanoemulsion through nose gets lots more to the brain (mice) – July 2020
- COVID-19 defeated 3x faster by 420,000 IU Vitamin D nanoemulsion – RCT Nov 12, 2020
- Bioavailability of nanoemulsion formulations of Vitamin D3 – Nov 2019
- Ulcerative Colitis significantly reduced by 480,000 IU loading dose of nano Vitamin D – RCT July 2019
- Vitamin D nanoemulsion etc. for fortification, pills, injections, topical and cancer – July 2019
- Vitamin D Cofactors in a nutshell
- Nanoemulsion Vitamin D may be a substantially better form has the following comparison
|50,000 IU powder in capsule |
Example Biotech Pharmacal
Example micro D3
per day for 10,000 IU
|4 cents||8 cents|
|IU per serving||50,000 IU = capsule||2,000 IU = drop|
|Servings if want average |
of 10,000 IU/day
|1 capsule |
per 5 days
|25 drops = 1 /4 teaspoon |
per 5 days
|Shelf life||1 year?||6 months?|
|Add to food/drink||Yes (powder)||possiblly|
|Apply to skin||No||Yes|
|Swish in mouth|
for fast response
|Yes if put powder in saliva|
or swish vitamin D water
|Availability to cell|
- better than bio-availability
|standard||perhaps 2X more|
- due to small size
or activation of Vitamin D Receptor
Download the PDF from VitaminDWiki
Table of Contents
extracted from 42 page PDF
Vitamin D (calciferols, Figure 2) is the name for the steroid hormonal precursors of calcitriol, a hormone that affects the resorption of calcium and phosphate from the intestine, regulating the levels of calcium and phosphorus in the blood, so it is important for strong and undamaged bones [67,74,94]. It is important for the proper functioning of the immune system (long-term deficiency is associated with respiratory infections and influenza). It is important for alleviating immunodermatological problems [20,21,67]. Vitamin D affects approx. 200 different chemical reactions in the body and is found in all types of human cells and in all human tissues. Structurally, vitamin D occurs in two modifications: vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol) . Vitamin D3 is produced in the skin by the action of sunlight (UVB) from provitamin 7-dehydrocholesterol. This synthesis covers 80% of the daily requirement. The amount of vitamin D produced is reduced by protective creams, frequent baths in hot water, dry skin of the elderly, large amounts of melanin in the skin, body envelopes, and air pollution. In food, vitamin D3 is found in fish oil, liver, egg yolk, and milk. The recommended dose for adults is 2000-4000 IU per day with blood levels of 30-60 ng/mL. At least two-thirds of all people living in northern latitudes are deficient in vitamin D. In plants, the precursor of vitamin D2 is ergosterol ormorphine [67,74,94].
Maurya et al.  comprehensively overviewed present findings related to fortification of food products with vitamin D with emphasis on factors affecting its bioavailability and the application of various suitable microencapsulation techniques, including liposomes, SLNPs, NLCs, NEs, spray drying, etc., which can be used for this purpose. Vitamin D NE-based delivery systems fabricated by spontaneous emulsification, which could be used in food industry, were reported by Guttoff et al. . The NE-based delivery system was found to increase the in vitro bioavailability of vitamin D3 3.94-fold, and according to the in vivo test, in which vitamin D3 NE and vitamin D3 coarse emulsion were used, the application of the nanoformulation resulted in a ca. two-fold increase in 25-hydroxycholecalciferol in serum compared to the coarse emulsion (an increase of 73% vs. 36%) .
Vitamin D3 incorporated into the polymeric complex of carboxymethyl chitosan (CMCS) with soy protein isolate (SPI) showing sizes of 162-243 nm, zeta potentials ranging from -10 to -20 mV, and EE up to 96.8% exhibited a ca. two-fold lower release in SGF (42.3% vs. 86.1%) but a ca. 4.4-fold higher release in SIF (36.0% vs. 8.2%) compared to NPs fabricated using SPI . Spherical N,N-dimethylhexadecyl CMCS core-shell micelles with a positive charge (+50.7 mV) encapsulated vitamin D3 with 53.2% EE resulting in its improved solubility. These core-shell micelles released vitamin D3 at first rapidly; later its sustained release was observed . Nanocomplexes prepared from ovalbumin, high-methoxylated pectin and encapsulated vitamin D3 showing high EE of 96.37% were characterized with electrostatic interactions, hydrogen bonding, and hydrophobic interactions among the three constituents and released only a small amount of vitamin D3 in SGF, while a large amount in SIF suggesting their potential to be used in food applications . High amylose starch nanocarriers with particle sizes of 14.2-31.8 nm and a negative surface charge loaded with vitamin D3 and achieving 37.06-78.11% EE that were investigated for food fortification using milk as a model food supplementing Ca improved the bioavailability of vitamin D3 and masked the after taste, suggesting their potential to be used for the fortification of food with vitamin D3 . In oil-in-water (O/W) Pickering emulsions stabilized by nanofibrillated cellulose (NFC; diameter: ca. 60 nm, length: several micrometers) encapsulating vitamin D3 containing 0.01% w/w vitamin D3,9.99% w/w soybean oil, 0.10-0.70% w/w NFC as emulsifier at phosphate buffer of pH 7, the extent of lipid digestion and vitamin bioavailability decreased with increasing NFC concentration . Mitbumrung et al.  encapsulated vitamin D3 in 10% wt soybean O/W Pickering emulsions stabilized by NFC or whey protein isolate (WPI) providing good stability to the emulsions via a combination of steric and electrostatic repulsion, where emulsions properties and EE were not affected by heating or ionic strength, and at highly acidic conditions (pH 2), particle size increased and EE showed a decrease. By an increase in NFC or WPI concentration, the stability and EE of the emulsions was improved and the encapsulated vitamin was effectively protected against environmental stresses occurring in industrial food production (e.g., pH changes, salt addition, and thermal processing).
The application of digestible oil (DO), indigestible oil (IO), or their mixture affected both the lipid digestion rate and the bioavailability of vitamin D3 encapsulated in NEs. The highest lipid digestion rate and vitamin bioavailability were observed with NEs using DO, the lowest one with those using IO, while comparable results were obtained with oil mixture (OM) consisting of 1:1 DO:IO mixed before homogenization and a 1:1 mixture consisting of DO and IO NEs mixed after homogenization. The maximum amount of vitamin D3 was estimated after ca. 30 min, and then its level showed a decrease during the following 24 h, which could be connected with an initial solubilization of the vitamin within the mixed micelles and following precipitation during prolonged incubation . From O/W NEs prepared using various oils and natural surfactant, quillaja saponin, encapsulating vitamin D3, the release of free fatty acids during lipid digestion in a simulated gastrointestinal tract (GIT) model decreased as follows: medium chain triglycerides (MCT) > corn oil > fish oil > orange oil > mineral oil, while the bioavailability of vitamin D3 increased in following order MCT < mineral oil< orange oil < fish oil < corn oil suggesting that the greatest increase in vitamin D3 bioavailability can be obtained with NEs fabricated with long chain triglycerides (corn or fish oil) . By blending caprylic-/capric triglyceride and Kolliphor HS®15, vitamin D3 and sodium chloride in optimal ratio, Maurya and Aggarwal  prepared a formulation with encapsulated vitamin able to tolerate environmental stress conditions, and based on sensory evaluation it was found to be suitable for fortification of vitamin D3 in “Lassi”, a milk based beverage. Uncoated nanoliposomes loaded with vitamins D3 and K2 were fabricated using a novel, a semi continuous technique based on simil-microfluidic principles and covered with CS to enhance the mucoadhesiveness and the stability of the liposomal structures, whereby CS was tested as covering material. Such polymer-lipid hybrid NPs encapsulating the above-mentioned vitamins were characterized with improved stability, loading, and mucoadhesiveness, suggesting their potential to be used in nutraceutical applications .
Vitamin D3 was incorporated into an NLC consisting of Precirol® (glyceryl palmitostearate) as a solid lipid and octyl octanoate as a liquid lipid. The surface of these NLCs was coated with either Poloxamer 407 or Tween 20. Both of these surfactants prevented agglomeration during the homogenization process while increasing intestinal absorption of the entire formulation, suggesting that NLCs can be used as an excellent carrier to enrich beverages with vitamin D3 .
Berinoetal.studied the interaction of vitamin D3 with p-lactoglobulinathighvitamin/protein ratios and found that when 100 uM vitamin D3 and 20 uM p-lactoglobulin in 20 mM phosphate buffer at pH 7.0 were used, vitamin D3 interacted in the hydrophobic calix in the protein, and the binding of the vitamin caused conformational changes in the secondary p-lactoglobulin structure. With the increasing vitamin concentration, the proportion of bound vitamin increased likely due to a cooperative phenomenon and/or a stacking process. Moeller et al.  enriched low fat yoghurt by spray- and freeze-dried casein micelles loaded with vitamin D2 maintaining constant vitamin content in powders during 4 months of storage. Based on the results of an in vitro proteolysis, when 90% of the vitamin D2 encapsulated in dry casein micelles remained active compared to 67% of free vitamin D2, it was assumed that after proteolysis, the vitamin will be ultimately available in the lumen. Using the optimal loading of vitamin D3 into re-assembled casein micelles (1.38-1.46 mg/100 mg casein) performed at 4.9 mM PO43-, 4.0 mM citrate, and 26.1 mM Ca, more vitamin D3 was retained in the re-assembled casein micelles than in control powders during storage, however its loss after 21 days of refrigerated storage with light exposure was comparable with that of the control fortified milks suggesting that re-assembled casein micelles can improve vitamin D3 stability during dry storage . The highly protective effect of the re-assembled casein micelles against gastric degradation of vitamin D3 resulted in its four-fold higher bioavailability compared to the free vitamin D3 .
Vitamin D3 and potato protein co-assemblies formed in phosphate buffer at pH 2.5 provided transparent solutions that were able to significantly protect and reduce vitamin D3 losses during pasteurization. Testing performed under different storage conditions suggests that potato protein could be used as a good carrier of vitamin D3 and the entire stable formulation could be used to fortify clear beverages, other foods, and drink products with vitamin D3 .
Pea protein-stabilized NEs with particle sizes of 170-350 nm and zeta-potential of -25 mV, which were characterized with good stability and the high EE of D vitamin (94-96%) exhibited considerably higher cellular uptake than emulsions fabricated using a combination of protein and lecithin, the cellular uptake of NEs with particle sizes of 233 nm being higher than that observed with NEs of 350 nm. Evently the transport efficiency of vitamin D in NEs with smaller particle sizes across Caco-2 cell was 5.3-fold greater than that of free vitamin D suspension, suggesting that pea protein could be considered as an effective emulsifier for fabrication of food NEs ensuring the improved bioavailability of vitamin D . Pea protein isolate (PPI), the function properties of which were modified using pH-shifting and sonication combined treatment, was applied to prepare NEs encapsulating vitamin D3. The NEs ensured good protection of vitamin D3 against UV radiation, were stable during 30-day storage, and showed ameliorated antioxidant activity as well as markedly higher recovery of vitamin D3 in micelles through in vitro digestion, suggesting that such NEs could be used for protection and delivery of nutraceuticals in foods . The application of vitamin D3 encapsulated in PPI NE at the dose of 81 daily to vitamin D deficient rats for one week resulted in higher serum 25-hydroxycholecalciferol levels compared to the control as well as in changes in serum parathyroid hormone, Ca, P, and alkaline phosphatase levels as compared to the controls. Hence, vitamin D3 encapsulated in PPI-based NEs improved its absorption and restored its status and biomarkers of bone resorption in vitamin D deficient rats .
Salvia-Trujillo et al.  investigated the impact of the initial lipid droplet size on the in vitro bioavailability and in vivo absorption of vitamin D2 encapsulated in O/W NE. The in vitro studies, in which vitamin D2-loaded lipid droplets were passed through a simulated GIT, showed the highest
bioavailability of the vitamin with the emulsions containing the smallest droplets, because they were digested more rapidly than larger ones and were able to form quickly mixed micelles in the small intestine capable to solubilize the lipophilic vitamin. On the other hand, in the in vivo rat feeding studies, the highest absorption of vitamin D2 was observed with NEs containing the largest droplets. This discrepancy could be connected with the fact that the simulated GIT cannot precisely reflect the complexity of a real GIT and by the applied in vivo approach, the changes in vitamin levels in the blood were not monitored over time.
Using mixed surfactant (Tween 80 and soya lecithin), vitamin D NEs were fabricated by ultrasonic homogenization showing droplet sizes of 140.15 nm and 155.5 nm after 2 months storage at 4 and 25 〇C, respectively; after 30 days of storage at 4 and 25 〇C, the NEs retained 74.4 ± 1.2 and 55.3 ± 2.1%〇 of vitamin D, suggesting their suitability to be used in food and beverages . The optimized vitamin D NEs fabricated by Mehmood et al.  using ultrasonication and lecithin and Tween 80 at a ratio 2:3 showed the size of 112.36 ± 3.6 nm and the vitamin D retention of 76.65 ± 1.7%〇. The higher release of vitamin D3 under simulated intestinal condition was observed from NEs co-encapsulating vitamin D3 and saffron petals' bioactive compounds, which were stabilized with basil seed gum and prepared using high pressure and ultrasound compared to those fabricated using WPC and Tween 80 emulsifiers . The investigation of a series of 2 wt% O/W emulsions containing different initial levels and locations of CS NPs and Tween 80 with encapsulated vitamin D3 showed that the NEs stabilized with Tween 8 exhibited 30% higher lipid digestion and 45% higher vitamin D3 bioavailability than those prepared with CS NPs, and the resulting effect depended on the applied ratio of CS NPs and Tween 80. It can be assumed that a layer of CS NPs limit the lipase to reach the lipid phase, the significant aggregation of droplets coated with CS NPs reduced the area of lipids, which is accessible to the lipase, and the positively charged CS NPs bound to anionic bile acids, fatty acids, or lipase. While the slowing of lipid digestion by CS NPs would be favorable at application in high-satiety foods, the reduced bioavailability of vitamin D is unfavorable . O/W NEs prepared using Tween 20, soybean lecithin, and their mixtures as emulsifiers and soybean oil or mixtures of the oil with cocoa butter as a dispersed oil phase using high pressure homogenization, showing oil droplets encapsulating vitamin D3 with average diameters <200 nm, maintained physical stability for several weeks. In systems stabilized by Tweens, partial vitamin's embedment in the interface of NEs was observed. The whole-fat milk fortified with vitamin D3 enriched NEs remained stable to particle aggregation and gravitational separation for at least 10 days .
Leaving aside the above mentioned combined nanoformulation of vitamin D with vitamin C , it appears that the described nanoencapsulation of vitamin D into casein , micelles and their application to yoghurt  has the greatest benefit for immunity of the nanoformulations
ALG (alginate); AST (astaxanthin); (3-Car (p-carotene); p-CD (p-cyclodextrin); CFU (colony-forming unit); CLPs (colloidal lipid particles); CMC (carboxymethyl cellulose); CMCS (carboxymethyl chitosan); CNC (central nervous system); CoQ10 (Coenzyme Q10); COS (chitooligosaccharide); COVID-19 (coronavirus disease caused by the SARS-CoV-2 virus); CS (chitosan); CUR (curcumin); DHA (docosahexaenoic acid); DO (digestible oil); DPPH (2,2-diphenyl-1-picrylhydrazyl); EE (encapsulation efficiency); EFSA (European Food Safety Authority); EGCG (epigallocatechin-3-gallate); FA (folic acid); FFAs (free fatty acids); FJM (fruitjuice-milk); GA (gum arabic); GABA (gamma-aminobutyric acid); GI (gastrointestinal); GIT (gastrointestinal tract); HA (hyaluronic acid); IFNy (interferon gamma); IL-6 (interleukin-6); IO (indigestible oil); a-LA (a-linolenic acid); LbL (layer-by-layer); LDHs (layered double hydroxides); LNCPs (lipid nanocapsules); MAPK (mitogen activated protein kinase); MCT (medium chain triglycerides); MDX (maltodextrin); MPs (microparticles); Na-ALG (sodium alginate); NaOl (sodium oleate); NE (nanoemulsion); NFC (nanofibrillated cellulose); NLCs (nanostructured lipid carriers); NPs (nanoparticles); ovalbumin (OVA); O/W (oil-in-water); PBMCs (peripheral blood mononuclear cells); PC (phosphatidylcholine); PCR (polymerase chain reaction); PEG (polyethylene glycol); PPI (pea protein isolate); PUFA (polyunsaturated fatty acid); PWP (polymerized whey protein); QR (quercetin); RebA (rebaudioside A); RES (resveratrol); ROS (reactive oxygen species); SGF (simulated gastric fluid); SIF (simulated intestinal fluid); SLNPs (solid lipid nanoparticles); SPI (soy protein isolate); TNFa (tumor necrosis factor alpha); a-Toc (a-tocopherol); W/O (water-in-oil); WP (whey protein); WPC (whey protein concentrate); WPI (whey protein isolate); ZX (zeaxanthin).
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