Elsevier

Carbohydrate Polymers

Volume 294, 15 October 2022, 119779
Carbohydrate Polymers

Amphiphilic nano-delivery system based on modified-chitosan and ovalbumin: Delivery and stability in simulated digestion

https://doi.org/10.1016/j.carbpol.2022.119779Get rights and content

Abstract

Nano-delivery systems play an important role in the development of nutritional supplements due to their efficient encapsulation and delivery properties for nutrients. Herein, we prepared protein-polysaccharide nanoparticles as a novel amphiphilic nano-delivery system based on gallic acid modified chitosan (GCS) and ovalbumin (OVA) by pH-driven and calcium ion crosslinking. The nanoparticles loaded with hydrophilic riboflavin (Rib) and hydrophobic quercetin (Que) as nutrient models were abbreviated as GCS-OVA-Rib NPs and GCS-OVA-Que NPs, respectively. Their encapsulation efficiencies for Rib and Que. were 66.36 % and 96.61 %, respectively. In addition, GCS-OVA-Rib NPs and GCS-OVA-Que NPs showed antioxidant activity as well as good stability and delivery capacity for Rib and Que. in simulated digestion with release ratios of 78.38 % and 84.15 %, respectively. More importantly, GCS-OVA-Rib/Que. NPs performed good biocompatibility for further applications. Overall, this work provides some useful insights for the design of novel amphiphilic nano-delivery systems based on polysaccharides and proteins.

Introduction

With the development of society and growing pursuit of health, people pay more attention to their health and nutrition, which are closely associated with bioactive nutrients (Tomadoni et al., 2020; Wang et al., 2021). Deficiency of these bioactive substances causes malnutrition and deficiency diseases, and many of them have been reported to make significant effects on diseases such as cancer, diabetes and depression. However, bioactive nutrients tend to be unstable and sensitive to light, oxygen, pH and gastrointestinal environment, limiting their application in practical production, affecting the biological accessibility and impairing the therapeutic effect. Fortunately, nano-delivery systems like nanoparticles show remarkable capacity to overcome these challenges and can improve the stability and bioavailability of bioactive nutrients (Chen et al., 2022; Dong et al., 2021; Li et al., 2022; Liang et al., 2019; Ma et al., 2020; Wang et al., 2022; Zhang, Cai, et al., 2021). Recently, many researchers have paid attention to nano-delivery systems for delivering hydrophilic or hydrophobic nutrients, but there are few studies on the nano-delivery system suitable for delivering both nutrients simultaneously.

Nano-delivery systems based on protein have been testified to possess high encapsulation efficiency for both hydrophilic nutrients (such as epigallocatechin gallate, crude extract of polyphenols and water-soluble vitamin) and hydrophobic nutrients (such as curcumin, β-carotene, quercetin, and lipid-soluble vitamins) due to the amphipathy of protein (Chai et al., 2013; Liu et al., 2022; Manjari et al., 2020; Wang et al., 2016; Yan et al., 2019; Yan et al., 2020; Zhang et al., 2022). Moreover, protein-based nano-delivery systems are biocompatible and can be fabricated by self-assembly without adding chemicals. Ovalbumin (OVA) is a main protein in egg white that can efficiently encapsulate and deliver bioactive nutrients including both hydrophilic and hydrophobic nutrients because of its amphipathy (Tomadoni et al., 2020; Visentini et al., 2019). Meanwhile, OVA can meet the nutritional requirements of human better than other commonly used plant proteins due to presence of useful amino acids (Rehault-Godbert et al., 2019). In addition, many studies have successfully applied OVA to nano-delivery systems. Hu et al. (2021) produced ovalbumin nanogels as stable delivery vehicles for curcumin achieving high encapsulation efficiency and sustained release in gastrointestinal condition; Zhang, Wang, et al. (2021) constructed nano-carriers based on OVA and propylene glycol alginate to improve the stability of anthocyanin; Zeng et al. (2021) synthesized OVA-pullulan nanogels to deliver curcumin. These studies successfully promoted the delivery application of OVA and identified its great potential for the delivery of hydrophobic and hydrophilic nutrients. However, there are few reports about the same delivery system based on OVA simultaneously studying both hydrophobic and hydrophilic nutrients. Therefore, it makes significant sense to design a novel nano-delivery system based on OVA simultaneously involving both hydrophobic and hydrophilic nutrients.

Considering nano-delivery system based on single protein tends to be unstable and easily broken down by pepsin so that embedded nutrients cannot be released in intestine where nutrients are absorbed (Patra et al., 2019), we turned our attention to polysaccharide-protein nano-delivery system. Advantages of polysaccharide-protein nano-delivery systems including stability enhancement, protein aggregation preventability (Thongkaew et al., 2014; Wei et al., 2015; Xia et al., 2015) and burst release retardation (Ru et al., 2010) have been thoroughly validated and reviewed. As an electropositive and widely applied polysaccharide (Parhi, 2020; Wei & Gao, 2016; Yang et al., 2019; Zou et al., 2016), chitosan (CS) stands out because it can assemble with protein avoiding harmful chemical agents or Maillard reactions unsuitable for heat-sensitive nutrients (Ding et al., 2019; Gupta, Jabrail, 2006a; Gupta, Jabrail, 2006b; Gupta & Jabrail, 2007; Hu et al., 2011; Teng et al., 2013; Zhang, Zeng, et al., 2021). However, its poor water solubility is a severe obstacle limiting the application in delivery system (Ivanova & Yaneva, 2020; Quan et al., 2021; Yang, Liu, et al., 2021). Modified chitosan with great water solubility while maintaining its original property is expected to form nano-complex with protein as novel amphiphilic delivery system, which can be used in food industry or pharmaceutical industry. Therefore, it is urgent need to modify chitosan through green method for its application in delivery systems.

Aspired by the above, for the first time, we designed a novel nano-delivery system based on OVA and gallic acid modified-chitosan through green and safe methods involving pH-driven self-assembly and calcium ion crosslinking. GCS-OVA-Rib NPs and GCS-OVA-Que NPs were fabricated with hydrophilic riboflavin (Rib) and hydrophobic quercetin (Que) as model substances. We hypothesized that OVA-GCS nano-delivery system in the form of nanoparticles could be successfully prepared through the method and efficiently delivery both hydrophilic (Rib) and hydrophobic (Que) drugs simultaneously. Therefore, we further studied whether the OVA-GCS nano-delivery system could maintain antioxidant property from GCS, perform great biocompatibility, especially control drug release and improve drug stability in gastric intestinal fluid. This study provides a new way for green synthesis of amphiphilic nano-delivery systems based on proteins and polysaccharides, making some effects on the development of nutritional supplements and drugs.

Section snippets

Materials

Chitosan (CS, deacetylation degree of 95 %, viscosity is 100–200 mPa·s), gallic acid (GA), trypsin originated from pig, 2,2-Diphenyl-1-picryl hydrazyl (DPPH) and 2,2′-Azino-bis, 3-ethylbenzothiazoline-6-sulphonic acid (ABTS) were obtained from Aladdin Biochemical Technology Co., ltd. (Shanghai, China). Ovalbumin (OVA) and Folin-Ciocalteu's phenol reagent were purchased from Beijing Solorbio Science & Technology. Calcium chloride dihydrate (CaCl2•2H2O), sodium chloride (NaCl), sodium hydroxide

Fabrication and characterization of gallic acid modified chitosan (GCS)

As shown in Fig. 1A, CS reacted with the mixture of ascorbic acid and hydrogen peroxide to produce GCS. UV, FTIR, and XRD characterization were applied to verify the synthesis mechanism and structure. In the UV spectra of CS, GA and GCS (Fig. 1B), CS showed no absorbance peak in the range of 200–800 nm, while there were two absorbance peaks in GA at 219 nm and 266 nm that were originated from the π system of its benzene ring (Xie et al., 2014). GCS also exhibited two peaks at 210 nm and 259 nm,

Conclusion

In summary, we designed a novel amphiphilic nano-delivery system based on OVA and GCS, which aimed at great delivery property for both hydrophilic Rib and hydrophobic Que. We successfully demonstrated that amphiphilic OVA-GCS nano-delivery system with high encapsulation capacity and slow-release property for Rib and Que. in SGF and SIF could be prepared in the form of nanoparticles by pH driven self-assembly and calcium ion crosslinking. The amphiphilic property suggests this delivery system

CRediT authorship contribution statement

Lihua Li: Conceptualization, Methodology, Validation, Formal analysis, Investigation, Writing – original draft. Xinyu Sun: Supervision, Writing – review & editing. Hui Zhang: Writing – review & editing. Mengna Dong: Writing – review & editing. Jiao Wang: Writing – review & editing. Shuang Zhao: Writing – review & editing. Minghui Shang: Writing – review & editing. Xin Wang: Writing – review & editing. Hui Zhangsun: Writing – review & editing. Li Wang: Conceptualization, Writing – review &

Declaration of competing interest

The authors declare no conflict of interest.

Acknowledgments

This work was supported by Talented Program (A279021724) and Chinese University Scientific Fund (No.2452021123).

References (71)

  • Q. Hu et al.

    Polyphenol-chitosan conjugates: Synthesis, characterization, and applications

    Carbohydrate Polymers

    (2016)
  • Q. Hu et al.

    Formation of redispersible polyelectrolyte complex nanoparticles from gallic acid-chitosan conjugate and gum arabic

    International Journal of Biological Macromolecules

    (2016)
  • L. Levy et al.

    The effect of gallic acid interactions with iron-coated clay on surface redox reactivity

    Water Research

    (2020)
  • J. Li et al.

    Characterization of calcium alginate/ deacetylated konjac glucomannan blend films prepared by Ca2+ crosslinking and deacetylation

    Food Hydrocolloids

    (2018)
  • S. Li et al.

    N-trimethyl chitosan coated targeting nanoparticles improve the oral bioavailability and antioxidant activity of vitexin

    Carbohydrate Polymers

    (2022)
  • X. Liang et al.

    Advances in research on bioactivity, metabolism, stability and delivery systems of lycopene

    Trends in Food Science & Technology

    (2019)
  • F. Liu et al.

    Tailoring the properties of double-crosslinked emulsion gels using structural design principles: Physical characteristics, stability, and delivery of lycopene

    Biomaterials

    (2022)
  • J. Liu et al.

    Preparation, characterization and antioxidant activity of phenolic acids grafted carboxymethyl chitosan

    International Journal of Biological Macromolecules

    (2013)
  • Y. Liu et al.

    Tyrosinase-mediated grafting and crosslinking of natural phenols confers functional properties to chitosan

    Biochemical Engineering Journal

    (2014)
  • M.S. Manjari et al.

    Highly biocompatible novel polyphenol cross-linked collagen scaffold for potential tissue engineering applications

    Reactive and Functional Polymers

    (2020)
  • R. Meng et al.

    Preparation and characterization of zein/carboxymethyl dextrin nanoparticles to encapsulate curcumin: Physicochemical stability, antioxidant activity and controlled release properties

    Food Chemistry

    (2021)
  • S. Omidi et al.

    Modification of chitosan and chitosan nanoparticle by long chain pyridinium compounds: Synthesis, characterization, antibacterial, and antioxidant activities

    Carbohydrate Polymers

    (2019)
  • W. Tan et al.

    Synthesis and antioxidant action of chitosan derivatives with amino-containing groups via azide-alkyne click reaction and N-methylation

    Carbohydrate Polymers

    (2018)
  • Z. Teng et al.

    Carboxymethyl chitosan-soy protein complex nanoparticles for the encapsulation and controlled release of vitamin D(3)

    Food Chemistry

    (2013)
  • C. Thongkaew et al.

    Polyphenol interactions with whey protein isolate and whey protein isolate–pectin coacervates

    Food Hydrocolloids

    (2014)
  • B. Tomadoni et al.

    Self-assembled proteins for food applications: A review

    Trends in Food Science & Technology

    (2020)
  • F.F. Visentini et al.

    Self-assembled nanoparticles from heat treated ovalbumin as nanocarriers for polyunsaturated fatty acids

    Food Hydrocolloids

    (2019)
  • L. Wang et al.

    Fucoxanthin-loaded nanoparticles composed of gliadin and chondroitin sulfate: Synthesis, characterization and stability

    Food Chemistry

    (2022)
  • W. Wang et al.

    Quercetagetin loaded in soy protein isolate–κ-carrageenan complex: Fabrication mechanism and protective effect

    Food Research International

    (2016)
  • X. Wang et al.

    Fabrication and characterization of zein-tea polyphenols-pectin ternary complex nanoparticles as an effective hyperoside delivery system: Formation mechanism, physicochemical stability, and in vitro release property

    Food Chemistry

    (2021)
  • Z. Wei et al.

    Physicochemical properties of β-carotene bilayer emulsions coated by milk proteins and chitosan–EGCG conjugates

    Food Hydrocolloids

    (2016)
  • Z. Wei et al.

    Evaluation of structural and functional properties of protein–EGCG complexes and their ability of stabilizing a model β-carotene emulsion

    Food Hydrocolloids

    (2015)
  • S. Xia et al.

    Glycosylation of bovine serum albumin via maillard reaction prevents epigallocatechin-3-gallate-induced protein aggregation

    Food Hydrocolloids

    (2015)
  • W. Yan et al.

    Corn fiber gum-soybean protein isolate double network hydrogel as oral delivery vehicles for thermosensitive bioactive compounds

    Food Hydrocolloids

    (2020)
  • J. Yang et al.

    Advanced applications of chitosan-based hydrogels: From biosensors to intelligent food packaging system

    Trends in Food Science & Technology

    (2021)
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