Fabrication and characterization of PVA/WPI nanofibers containing probiotics using electrospinning technique

Document Type : Research Paper


1 Department of Food Hygiene and Aquaculture, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

2 Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 Departments of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad, University of Medical Sciences, Mashhad, Iran


Objective(s): This study aimed to evaluate the viability of encapsulated probiotics using electrospinning technique. Specifically, the study focused on polyvinyl alcohol-whey protein isolate nanofibers (PVA/WPI) containing Bifidobacterium bifidum. These nanofibers have potential applications in active food packaging to improve food safety and extend shelf life, as well as in medical and pharmaceutical fields.
Materials and Methods: PVA/WPI nanofibers were electrospun in varying ratios (ranging from 100:00 to 50:50) and evaluated for their morphology, mechanical properties, FT-IR and DSC characteristics. B. bifidum was also encapsulated in the optimized PVA/WPI nanofibers to assess their encapsulation efficiency and viability, and the antimicrobial properties of the nanofibers were determined using the disk diffusion method.
Results: All prepared nanofibers displayed a diameter range of 186.42-612.5 nm, with an inverse relationship between WPI ratio and nanofiber diameter. The PVA/WPI nanofiber with a ratio of 60:40 was found to be the most favorable. DSC analysis showed that adding WPI decreased thermal stability, and the enthalpy of endothermic peaks decreased in nanofibers containing B. bifidum. Mechanical evaluation revealed that adding WPI reduced tensile strength and elongation at break, without significant effects from B. bifidum (P>0.05). Bacterial encapsulation efficiency was 80.58%. Probiotic nanofibers exhibited antimicrobial properties against Listeria monocytogenes (11.00±0.37 mm) and Escherichia coli (9.71±0.06 mm).
Conclusion: According to the obtained results, the optimized PVA/WPI nanofiber (60:40) contained suitable morphological, mechanical and thermal characteristics with the highest encapsulation efficiency in regards to B. bifidum (>80%). Probiotics-containing PVA/WPI nanofibers are a suitable platform for medical applications and food industry packaging due to their antimicrobial properties.


1. Oak SJ, Jha R. The effects of probiotics in lactose intolerance: a systematic review. Crit Rev Food Sci Nutr. 2019;59(11):1675-1683.
2.    Shu G, He Y, Chen L, Song Y, Meng J, Chen H. Microencapsulation of Bifidobacterium bifidum BB01 by xanthan–chitosan: Preparation and its stability in pure milk. Artif Cells Nanomed Biotechnol. 2018;46(sup1):588-596.
3.    Fujiya M, Ueno N, Kohgo Y. Probiotic treatments for induction and maintenance of remission in inflammatory bowel diseases: a meta-analysis of randomized controlled trials. Clin J Gastroenterol. 2014;7(1):1-3.
4.    Mirhadi Zadi, T, Mohsenzadeh, M, Ghahramani Seno, MM. Molecular Isolation, Probiotic Property, and Bacteriocin Production of Enterococcus faecium (TM81) and Lactobacillus curvatus (TM51) with Anti-Listerial Activity in Native Dairy Products of Iran. Iran J Chem Chem Eng. 2021; 40(4):1346-1363. 
5.    Rahimifard N, Moghni M. Naseri. Evaluation and comparison of three antimicrobial activity methods using bifidobacteria bifidum and bifidobacteria infantis as probiotic bacteria against salmonella enterica serotype enteritidis. J Bacteriol Mycol. 2016;2(3):61-64.
6.    Afzaal M, Saeed F, Ateeq H, Ahmed A, Ahmad A, Tufail T, et al. Encapsulation of Bifidobacterium bifidum by internal gelation method to access the viability in cheddar cheese and under simulated gastrointestinal conditions. Food Sci Nutr. 2020;8(6):2739-2747.
7.    Liu H, Gong J, Chabot D, Miller SS, Cui SW, Zhong F, et al. Improved survival of Lactobacillus zeae LB1 in a spray dried alginate-protein matrix. Food Hydrocoll. 2018;78:100-108.
8.    Chávarri M, Marañón I, Villarán MC. Encapsulation technology to protect probiotic bacteria.InProbiotics 2012. IntechOpen.
9.    Çanga EM, Dudak FC. Improved digestive stability of probiotics encapsulated within poly (vinyl alcohol)/cellulose acetate hybrid fibers. Carbohydr Polym. 2021;264:117990.
10.    Fahami A, Fathi M. Fabrication and characterization of novel nanofibers from cress seed mucilage for food applications. J Appl Polym Sci. 2018;135(6):45811.
11.    Ramakrishna S. An introduction to electrospinning and nanofibers. World Scientific. 2005.
12.    Ma J, Xu C, Yu H, Feng Z, Yu W, Gu L, et al. Electro-encapsulation of probiotics in gum Arabic-pullulan blend nanofibres using electrospinning technology. Food Hydrocoll. 2021;111:106381.
13.    Feng K, Huang RM, Wu RQ, Wei YS, Zong MH, Linhardt RJ, et al. A novel route for double-layered encapsulation of probiotics with improved viability under adverse conditions. Food chem. 2020;310:125977.
14.    Mojaveri SJ, Hosseini SF, Gharsallaoui A. Viability improvement of Bifidobacterium animalis Bb12 by encapsulation in chitosan/poly (vinyl alcohol) hybrid electrospun fiber mats. Carbohydr Polym. 2020;241: 116278.
15.    Santos C, Silva CJ, Büttel Z, Guimarães R, Pereira SB, Tamagnini P, et al. Preparation and characterization of polysaccharides/PVA blend nanofibrous membranes by electrospinning method. Carbohydr polym. 2014;99:584-592.
16.    Badr K, Ahmed ZS, El-Gamal M. Assessment of antimicrobial activity of whey protein films incorporated with biocide plant-based essential oils. J Appl Sci Res. 2013;9(4):2811-2818.
17.    Tatlisu NB, Yilmaz MT, Arici M. Fabrication and characterization of thymol-loaded nanofiber mats as a novel antimould surface material for coating cheese surface. Food Packag Shelf Life. 2019;21:100347.
18.    Azevedo VM, Dias MV, de Siqueira Elias HH, Fukushima KL, Silva EK, Carneiro JD, et al. Effect of whey protein isolate films incorporated with montmorillonite and citric acid on the preservation of fresh-cut apples. Food Res Int. 2018; 107:306-313.
19.    Cordeiro BF, Oliveira ER, Da Silva SH, Savassi BM, Acurcio LB, Lemos L, et al. Whey protein isolate-supplemented beverage, fermented by Lactobacillus casei BL23 and Propionibacterium freudenreichii 138, in the prevention of mucositis in mice. Front Microbiol. 2018;9:2035.
20.    Duman D, Karadag A. Inulin added electrospun composite nanofibres by electrospinning for the encapsulation of probiotics: characterisation and assessment of viability during storage and simulated gastrointestinal digestion. Int J of Food Sci Technol. 2021;56(2):927-935.
21.    Wang WB, Clapper JC. Antibacterial Activity of Electrospun Polyacrylonitrile Copper Nanoparticle Nanofibers on Antibiotic Resistant Pathogens and Methicillin Resistant Staphylococcus aureus (MRSA). Nanomaterials. 2022; 12(13):2139.
22.    Colin-Orozco J, Zapata-Torres M, Rodriguez-Gattorno G, Pedroza-Islas R. Properties of poly (ethylene oxide)/whey protein isolate nanofibers prepared by electrospinning. Food Biophys. 2015;10(2):134-144.
23.    Sharma P, Trivedi N, Gat Y. Development of functional fermented whey–oat-based product using probiotic bacteria. 3 Biotech. 2017;7(4):1-8.
24.    Lugo A. C. V. Understanding the Role of Poly (ethylene oxide) in the Electrospinning of Whey Protein Isolate Fibers (Doctoral dissertation, University of Guelph). 2012.
25.    Diep E, Schiffman JD. Encapsulating bacteria in alginate-based electrospun nanofibers. Biomater Sci. 2021;9(12): 4364-4373.
26.    Škrlec K, Zupančič Š, Mihevc SP, Kocbek P, Kristl J, Berlec A. Development of electrospun nanofibers that enable high loading and long-term viability of probiotics. Eur J Pharm Biopharm. 2019;136:108-119.
27.    Keramat M, Esteghlal S, Safari J, Golmakani MT, Khalesi M. Fabrication of electrospun persian Gum/Poly (Vinyl Alcohol) and whey protein Isolate/Poly (vinyl alcohol) nanofibers incorporated with Oliveria decumbens Vent. essential oil. Nanosci Nanotechnol. 2019;9(3):371-380.
28.    Vega‐Lugo AC, Lim LT. Effects of poly (ethylene oxide) and pH on the electrospinning of whey protein isolate. J Polym Sci B Polym Phys. 2012;50(16):1188-1197.
29.    Ceylan Z, Meral R, Karakaş CY, Dertli E, Yilmaz MT. A novel strategy for probiotic bacteria: Ensuring microbial stability of fish fillets using characterized probiotic bacteria-loaded nanofibers. Innov Food Sci Emerg Technol. 2018;48:212-218.
30.    Yilmaz MT, Taylan O, Karakas CY, Dertli E. An alternative way to encapsulate probiotics within electrospun alginate nanofibers as monitored under simulated gastrointestinal conditions and in kefir. Carbohydr Polym. 2020;244: 116447.
31.    Dissanayake M, Vasiljevic T. Functional properties of whey proteins affected by heat treatment and hydrodynamic high-pressure shearing. J Dairy Sci. 2009;92(4):1387-1397.
32.    Patel MT, Kilara A, Huffman LM, Hewitt SA, Houlihan AV. Studies on whey protein concentrates. 1. Compositional and thermal properties. J Dairy Sci. 1990;73(6):1439-1449.
33.    Ucpinar Durmaz B, Aytac A. Effects of Polyol-Based Plasticizer Types and Concentration on the Properties of Polyvinyl Alcohol and Casein Blend Films. J Polym Environ. 2021;29(1):313-322.
34.    Sun Y, Liu Z, Zhang L, Wang X, Li L. Effects of plasticizer type and concentration on rheological, physico-mechanical and structural properties of chitosan/zein film. Int J Biol Macromol. 2020;143:334-340.
35.    Gialamas H, Zinoviadou KG, Biliaderis CG, Koutsoumanis KP. Development of a novel bioactive packaging based on the incorporation of Lactobacillus sakei into sodium-caseinate films for controlling Listeria monocytogenes in foods. Food Res Int. 2010;43(10):2402-2408.
36.    Ghalehjooghi HD, Tajik H, Shahbazi Y. Development and characterization of active packaging nanofiber mats based on gelatinsodium alginate containing probiotic microorganisms to improve the shelf-life and safety quality of silver carp fillets. Int J Food Microbiol. 2023;384: 109984.
37.    Kanmani P, Lim ST. Development and characterization of novel probiotic-residing pullulan/starch edible films. Food chem. 2013;141(2):1041-1049.
38.    Oliveira-Alcântara AV, Abreu AA, Gonçalves C, Fuciños P, Cerqueira MA, Gama FM, et al. Bacterial cellulose/cashew gum films as probiotic carriers. LWT-Food Sci Technol. 2020;130:109699.
39.    Kurt A, Toker OS, Tornuk F. Effect of xanthan and locust bean gum synergistic interaction on characteristics of biodegradable edible film. Int J Biol Macromol. 2017;102:1035-1044.
40.    Leuangsukrerk M, Phupoksakul T, Tananuwong K, Borompichaichartkul C, Janjarasskul T. Properties of konjac glucomannan–whey protein isolate blend films. LWT-Food Sci Technol. 2014;59(1):94-100.
41.    Marvdashti LM, Koocheki A, Yavarmanesh M. Alyssum homolocarpum seed gum-polyvinyl alcohol biodegradable composite film: Physicochemical, mechanical, thermal and barrier properties. Carbohydr Polym. 2017;155:280-293.
42.    Durmaz BU, Aytac A. Development and characterization of poly (vinyl alcohol) and casein blend films. Polym Int. 2019; 68(6):1140-1145.
43.    Aloui H, Baraket K, Sendon R, Silva AS, Khwaldia K. Development and characterization of novel composite glycerol-plasticized films based on sodium caseinate and lipid fraction of tomato pomace by-product. Int J Biol Macromol. 2019; 139:128-138.
44.    Fung WY, Yuen KH, Liong MT. Agrowaste-based nanofibers as a probiotic encapsulant: fabrication and characterization. J Agric Food Chem. 2011; 59(15):8140-8147.
45.    Hu D, Wang L. Physical and antibacterial properties of polyvinyl alcohol films reinforced with quaternized cellulose. J Appl Polym Sci. 2016;133(25).
46.    Chollakup R, Pongburoos S, Boonsong W, Khanoonkon N, Kongsin K, Sothornvit R, Sukyai P, et al. Antioxidant and antibacterial activities of cassava starch and whey protein blend films containing rambutan peel extract and cinnamon oil for active packaging. LWT-Food Sci Technol. 2020;130:109573.
47.    Wahbi W, Siam R, Kegere J, El-Mehalmey WA, Mamdouh W. Novel inulin electrospun composite nanofibers: prebiotic and antibacterial activities. ACS omega. 2020; 5(6):3006-3015.
48.    Gibson GR, Wang X. Regulatory effects of bifidobacteria on the growth of other colonic bacteria. J Appl Bacteriol. 1994;77(4):412-420.
49.    Lim H. J., Shin H. S. Antimicrobial and immunomodulatory effects of Bifidobacterium strains: A review. J Microbiol Biotechnol. 2020;1793-1800.