Farnaud, S.

Renshaw, D.

Somavarapu, S.

Zariwala, M.

WestminsterResearch

See	discussions,	stats,	and	author	profiles	for	this	publication	at:	https://www.researchgate.net/publication/282669679Novel	whey	protein	isolate	nanocarriers	for	oralmicronutrient	deliveryConference	Paper	·	September	2015CITATIONS0READS531	author:Mohammed	Gulrez	ZariwalaUniversity	of	Westminster29	PUBLICATIONS			151	CITATIONS			SEE	PROFILEAll	content	following	this	page	was	uploaded	by	Mohammed	Gulrez	Zariwala	on	18	March	2016.The	user	has	requested	enhancement	of	the	downloaded	file.Novel whey protein isolate nanocarriers for oral micronutrient delivery Shamarah Mathurin-Charles1, Peter Agala Owanaro1, Sebastien Farnaud1, Derek Renshaw2, Satyanarayana Somavarapu3 and M.Gulrez Zariwala1,21 Department of Life Sciences, University of Bedfordshire, Park Square, Luton, Bedford LU1 3JU, United Kingdom2 Faculty of Science and Technology, University of Westminster, 115 New Cavendish street, London W1W 6UW, United Kingdom3 Department of Pharmaceutics, UCL School of Pharmacy, 29-39 Brunswick Square, London, WC1N 1AX United KingdomMethodsWPI nanocarriers encapsulating FeSO4 (WPI-NC) were prepared using coldhomogenisation method. The mucoadhesive polysaccharide chitosan (CHI) wasadded to the aqueous phase to prepare chitosan coated nanocarriers (WPI-CHI-NC).Nanocarrier physiochemical characteristics were assessed by particle size, zetapotential and morphological analysis. Iron uptake from formulations wascompared by caco-2 cell uptake experiments using simulated GI fluid, withintracellular ferritin protein as a measure of iron absorption and pure FeSO4 asreference.Potential toxic effects of nanocarrier formulations on caco-2 cells were assessedby carrying out the colourimetric MTT assay incubating caco-2 cell monolayerswith formulations diluted at a final iron concentrations of 20, 50 and 100 µM (andequivalent volumes of corresponding blank iron-free nanocarriers).FeSO4 was used as a reference standard for quantitaive iron absorptionexperiments. Equivalent amounts of iron (20 µM) from each formulation wasadded to caco-2 cells cultured in six-well plates (n = 6 per sample) for 2 hours, theestimated physiological transit time through the duodenum, and cells harvestedafter 24 hours.Intracellular ferritin concentration was determined by ELISA. Ferritinconcentrations were then standardised against total protein concentration and ngferritin/mg protein considered an indice of iron absorption in Caco-2 cells. Data ispresented as mean ± SEM and difference between samples was analysed byANOVA followed by Tukey's post-hoc test using the PRISM software package.IntroductionIron deficiency is the most prevalent nutritional disorders worldwide (WHO).Ferrous sulphate (FeSO4) is the most common iron supplement/fortificant,however, it causes gastrointestinal (GI) side effects and has a poor sensoryprofile. Encapsulation approaches are used to overcome this, but poor absorptionis a limitation.Milk proteins are generally inexpensive, with an established safety andbiocompatibility profile and are widely used in the food industry for their nutritionaland functional properties.ResultsFig. 2. Particle size and morphological analysis was carried out by scanning electronmicroscopy as well as using a Malvern Zetasizer. All formulations were found to be within asubmicron size range (113.76 ± 7 nm – 125.92 ± 12 nm) favourable for intestinalpermeability. SEM analysis was in agreement with Zetasizer measurements, withnanocarrier particle size observed to be of nanoscale dimensions.Fig. 3. Caco-2 cell viability as assessed by MTT assay following 48 hour (a) and 72 hour (b)incubation with nanocarrier formulations containing increasing drug concentrations (n=6).Results demonstrate that the nanocarrier formulations did not exert any significant toxiceffects upon caco-2 cell viability.Fig. 4. Quantitative iron uptake from the nanocarrier formulations was compared by carryingout caco-2 cell uptake experiments with intracellular ferritin protein as a measure of ironabsorption. Caco-2 iron absorption from WPI-NC (88.96 ± 8.9 ng/mg) was only 23% ofFeSO4 control (374.61 ± 22.03 ng/mg). However, chitosan inclusion significantly increasedcellular iron absorption, with WPI-CHI-NC iron absorption (410.1ng/mg ± 24.77) 365.9 %higher than WPI-NC and 9.6 % higher than FeSO4 control (P ≤ 0.05).Summary Low level iron absorption from WPI-NC composed of WPI alone may potentially be attributed to poor nanocarrier integrity in the GI microenvironment. Chitosan inclusion leads to formation of robust hybrid protein-polysaccharide nanocarriers possessing greater membrane permeability resulting in highcellular iron delivery. Our results demonstrate the potential of WPI as a novel biomaterial for formulation of nanocarriers for micronutrient delivery.†Corresponding Author: Dr M.Gulrez Zariwala  Email: zariwam@westminster.ac.ukPoster No. 52Whey proteins isolate (WPI) isobtained by processing andfiltration of milk whey (Fig. 1.). Ithas a high protein content(97.5%) and is virtually lactose,carbohydrate and fat free.WPI is readily available andbiocompatible, and also hasphysiochemical properties suchas self assembly. Thesecharacteristics make it apromising material forencapsulation and formulation ofbioactive compounds.We describe for the first time preparation and characterisation of novelWPI composite nanocarriers for oral iron formulation and delivery.Fig. 1. Separation of dairy milk into proteinfractionsView publication stats

Novel whey protein isolate nanocarriers for oral micronutrient delivery

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Novel whey protein isolate nanocarriers for oral micronutrient delivery

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