Research Article
Nanofiber encapsulation of probiotic cultures via electrospinning: fabrication and quality compliance with ISO/IEC 17043 and ISO 22117 standards
Abstract
Probiotics offer numerous health benefits, including inhibiting pathogenic growth, supporting intestinal microbiota, and synthesizing essential biomolecules. However, their viability during storage remains a challenge due to sensitivity to environmental conditions. This study investigates the encapsulation of Lactobacillus rhamnosus and Lactobacillus acidophilus in polyvinyl alcohol (PVA) nanofibers via electrospinning to enhance stability and viability. Near-optimized electrospinning parameters, including solution concentration, voltage, and collector distance, were used to produce nanofibers, which were characterized using Field Emission Scanning Electron Microscopy (FESEM). The results showed non-uniform fiber diameter distributions, with 16 kV producing thicker fibers with an average diameter of 479.11 nm. Homogeneity assessment confirmed uniform probiotic distribution within the nanofibers, with a coefficient of variation of 5.3%. Storage stability tests at 4°C over 15 days were conducted following ISO/IEC 17043 and ISO 22117 standards. The findings demonstrated that encapsulation effectively preserved L. rhamnosus viability in 16LR/PVA nanofibers, whereas L. acidophilus exhibited reduced viability at both 10 kV and 16 kV.
Ethical Statement
Ethics committee approval is not required for this study.
Supporting Institution
This work was supported by the project numbered 2022HZDP011 by the scientific research project unit of Pamukkale University - Scientific Research Projects Coordinatorship.
Project Number
2022HZDP011 [ D29 ]
References
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- 12. Champagne CP, Gardner NJ (2008): Effect of storage in a fruit drink on subsequent survival of probiotic lactobacilli to gastro-intestinal stresses. Food Res Int, 41, 539–543.
- 13. Champagne CP, Ross RP, Saarela M (2011): Recommendations for the viability assessment of probiotics as concentrated cultures and in food matrices. Int J Food Microbiol, 149, 185–193.
- 14. Choo K, Ching YC, Chuah CH, et al (2016): Preparation and characterization of polyvinyl alcohol-chitosan composite films reinforced with cellulose nanofiber. Materials, 9, 644.
- 15. Coghetto CC, Brinques GB, Ayub MAZ (2016): Probiotics production and alternative encapsulation methodologies to improve their viabilities under adverse environmental conditions. Int J Food Sci Nutr, 67, 929–943.
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- 17. Deitzel JM, Kleinmeyer JD, Harris D, et al (2001): The effect of processing variables on the morphology of electrospun nanofibers and textiles. Polym, 42, 261–272.
- 18. Deng L, Zhang H (2020): Recent advances in probiotics encapsulation by electrospinning. ES Food Agrofor, 2, 3-12.
- 19. Eratte D, McKnight S, Gengenbach TR, et al (2015): Co-encapsulation and characterisation of omega-3 fatty acids and probiotic bacteria in whey protein isolate–gum Arabic complex coacervates. J Funct Foods, 19, 882–892.
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- 21. Feng K, Huangfu L, Liu C, et al (2023): Electrospinning and electrospraying: emerging techniques for probiotic stabilization and application. Polymer, 15, 2402.
- 22. Feng K, Wen P, Yang H, et al (2017): Enhancement of the antimicrobial activity of cinnamon essential oil-loaded electrospun nanofilm by the incorporation of lysozyme. RSC Adv, 7, 1572–1580.
- 23. Feng K, Zhai M, Zhang Y, et al (2018): Improved viability and thermal stability of the probiotics encapsulated in a novel electrospun fiber mat. J Agric Food Chem, 66, 10890–10897.
- 24. Fung W, Yuen K, Liong M (2011): Agrowaste-Based Nanofibers as a Probiotic Encapsulant: fabrication and characterization. J Agric Food Chem, 59, 8140–8147.
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- 30. ISO 4833-1 (2013): ISO. Available at https://www.iso.org/standard/53728.html. (Accessed May 5, 2024).
- 31. Khan MA, Hussain Z, Ali S, et al (2019): Fabrication of electrospun probiotic functionalized nanocomposite scaffolds for infection control and dermal burn healing in a mice model. ACS Biomater Sci Eng, 5, 6109-6116.
- 32. Kidoaki S, Kwon IK, Matsuda T (2005): Mesoscopic spatial designs of nano- and microfiber meshes for tissue-engineering matrix and scaffold based on newly devised multilayering and mixing electrospinning techniques. Biomaterials, 26, 37–46.
- 33. Koski A, Yim K, Shivkumar S (2004): Effect of molecular weight on fibrous PVA produced by electrospinning. Mater Lett, 58, 493–497.
- 34. Kumar M, Mohania D, Poddar D, et al (2009): A probiotic fermented milk prepared by mixed culture reduces pathogen shedding and alleviates disease signs in rats challenged with pathogens. Int J Probiotics Prebiotics, 4, 211–218.
- 35. Li Q, Jia Z, Yang Y, et al (2008): Preparation and properties of poly (vinyl alcohol) nanofibers by electrospinning. J Polym Eng. 28, 87-100.
- 36. Liu H, Cui SW, Chen M, et al (2019): Protective approaches and mechanisms of microencapsulation to the survival of probiotic bacteria during processing, storage and gastrointestinal digestion: a review. Crit Rev Food Sci Nutr, 59, 2863–2878.
- 37. Mitchell GR, Mohan SD, Davis FJ, et al (2015): Electrospinning: Principles, Practice and Possibilities. The Royal Society of Chemistry, Cambridge.
- 38. Mojaveri SJ, Hosseini SF, Gharsallaoui A (2020): Viability improvement of Bifidobacterium animalis Bb12 by encapsulation in chitosan/poly(vinyl alcohol) hybrid electrospun fiber mats. Carbohydr Polym, 241, 116278.
- 39. Mortazavian AM, Ehsani MR, Mousavi M, et al (2007): Effect of refrigerated storage temperature on the viability of probiotic microorganisms in yogurt. Int J Dairy Technol, 60, 123–127.
- 40. Nagy Z, Wagner I, Suhajda Á, et al (2014): Nanofibrous solid dosage form of living bacteria prepared by electrospinning. Express Polym Lett, 8, 352–361.
- 41. Njjar MS, Akduman Ç, Koluman A (2023): Antibakteriyel, kanama durdurucu ve yaralanma tespit sistemi içeren askeri operasyon kıyafeti. Savunma Bilim Derg, 2, 424–453.
- 42. Ricaurte L, Quintanilla-Carvajal MX (2019): Use of electrospinning technique to produce nanofibres for food industries: a perspective from regulations to characterisations. Trends Food Sci Technol, 85, 92–106.
- 43. Rwei S, Huang C (2012): Electrospinning PVA solution-rheology and morphology analyses. Fibers Polym, 13, 44–50.
- 44. Salalha W, Dror Y, Khalfin RL, et al (2004): Single-walled carbon nanotubes embedded in oriented polymeric nanofibers by electrospinning. Langmuir, 20, 9852–9855.
- 45. Sanz Y (2007): Ecological and functional implications of the acid-adaptation ability of Bifidobacterium: a way of selecting improved probiotic strains. Int Dairy J, 17, 1284–1289.
- 46. Simonič M, Slapničar Š, Trček J, et al (2023): Probiotic Lactobacillus paragasseri K7 nanofiber encapsulation using nozzle-free electrospinning. Appl Biochem Biotechnol, 195, 6768–6789.
- 47. Škrlec K, Zupančič Š, Mihevc SP, et al (2019): Development of electrospun nanofibers that enable high loading and long-term viability of probiotics. Eur J Pharm Biopharm, 136, 108–119.
- 48. Soukoulis C, Singh P, MacNaughtan W, et al (2016): Compositional and physicochemical factors governing the viability of Lactobacillus rhamnosus GG embedded in starch-protein based edible films. Food Hydrocoll, 52, 876–887.
- 49. Tsiouris CG, Tsiouri MG (2017): Human microflora, probiotics and wound healing. Wound Med, 19, 33–38.
- 50. Zavišić G, Ristić S, Petković B, et al (2023): Microbiological quality of probiotic products. Arhiv Za Farmaciju, 73, 17–34.
- 51. Ziabari M, Mottaghitalab V, Haghi AK (2010): A new approach for optimization of electrospun nanofiber formation process. Korean J Chem Eng, 27, 340–354.
Year 2025,
Volume: 72 Issue: 3, 323 - 333, 01.07.2025
Abstract
Project Number
2022HZDP011 [ D29 ]
References
- 1. Agarwal S, Greiner A (2011): On the way to clean and safe electrospinning—green electrospinning: emulsion and suspension electrospinning. Polym Adv Technol, 22, 372–378.
- 2. Akduman Ç, Morsümbül S, Kumbasar EPA (2019): The removal of reactive red 141 from wastewater: a study of dye adsorption capability of water-stable electrospun polyvinyl alcohol nanofibers. Autex Research Journal, 21, 20–31.
- 3. Amna T, Hassan MS, Pandeya DR, et al (2013): Classy non-wovens based on animate l. Gasseri-inanimate poly(vinyl alcohol): upstream application in food engineering. Appl Microbiol Biotechnol, 97, 4523–4531.
- 4. Anekella K, Orsat V (2013): Optimization of microencapsulation of probiotics in raspberry juice by spray drying. LWT- Food Sci Technol, 50, 17–24.
- 5. Atkins P, Stainback L (2022): What does your proficiency testing (PT) prove? a look at inorganic analyses, proficiency tests, and contamination and error. Spectroscopy, Part I, 9–14.
- 6. Ayutsede J, Gandhi M, Sukigara S, et al (2005): Regeneration of Bombyx mori silk by electrospinning. Part 3: characterization of electrospun nonwoven mat. Polymer, 46, 1625–1634.
- 7. Baker SC, Atkin N, Gunning PA, et al (2006): Characterisation of electrospun polystyrene scaffolds for three-dimensional in vitro biological studies. Biomaterials, 27, 3136–3146.
- 8. Barrientos S, Stojadinović O, Golinko MS, et al (2008): Perspective Article: Growth factors and cytokines in wound healing. Wound Rep Reg, 16, 585–601.
- 9. Bhushani JA, Anandharamakrishnan C (2014): Electrospinning and electrospraying techniques: potential food based applications. Trends Food Sci Technol, 38, 21–33.
- 10. Caramia G, Atzei A, Fanos V (2008): Probiotics and the skin. Clin Dermatol, 26, 4–11.
- 11. Ceylan Z, Meral R, Karakaş CY, et al (2018): A novel strategy for probiotic bacteria: ensuring microbial stability of fish fillets using characterized probiotic bacteria-loaded nanofibers. Innov Food Sci Emerg Technol, 48, 212–218.
- 12. Champagne CP, Gardner NJ (2008): Effect of storage in a fruit drink on subsequent survival of probiotic lactobacilli to gastro-intestinal stresses. Food Res Int, 41, 539–543.
- 13. Champagne CP, Ross RP, Saarela M (2011): Recommendations for the viability assessment of probiotics as concentrated cultures and in food matrices. Int J Food Microbiol, 149, 185–193.
- 14. Choo K, Ching YC, Chuah CH, et al (2016): Preparation and characterization of polyvinyl alcohol-chitosan composite films reinforced with cellulose nanofiber. Materials, 9, 644.
- 15. Coghetto CC, Brinques GB, Ayub MAZ (2016): Probiotics production and alternative encapsulation methodologies to improve their viabilities under adverse environmental conditions. Int J Food Sci Nutr, 67, 929–943.
- 16. De Mandal S, Hati S (2016): Microencapsulation of bacterial cells by emulsion technique for probiotic application. 273-279. In: EC Opara (Ed), Cell Microencapsulation: Methods and Protocols. Humana Press, New York.
- 17. Deitzel JM, Kleinmeyer JD, Harris D, et al (2001): The effect of processing variables on the morphology of electrospun nanofibers and textiles. Polym, 42, 261–272.
- 18. Deng L, Zhang H (2020): Recent advances in probiotics encapsulation by electrospinning. ES Food Agrofor, 2, 3-12.
- 19. Eratte D, McKnight S, Gengenbach TR, et al (2015): Co-encapsulation and characterisation of omega-3 fatty acids and probiotic bacteria in whey protein isolate–gum Arabic complex coacervates. J Funct Foods, 19, 882–892.
- 20. FAO (2006): Probiotics in food: Health and nutritional properties and guidelines for evaluation. Food and Agriculture Organization of the United Nations, Rome.
- 21. Feng K, Huangfu L, Liu C, et al (2023): Electrospinning and electrospraying: emerging techniques for probiotic stabilization and application. Polymer, 15, 2402.
- 22. Feng K, Wen P, Yang H, et al (2017): Enhancement of the antimicrobial activity of cinnamon essential oil-loaded electrospun nanofilm by the incorporation of lysozyme. RSC Adv, 7, 1572–1580.
- 23. Feng K, Zhai M, Zhang Y, et al (2018): Improved viability and thermal stability of the probiotics encapsulated in a novel electrospun fiber mat. J Agric Food Chem, 66, 10890–10897.
- 24. Fung W, Yuen K, Liong M (2011): Agrowaste-Based Nanofibers as a Probiotic Encapsulant: fabrication and characterization. J Agric Food Chem, 59, 8140–8147.
- 25. Gaaz TS, Sulong AB, Akhtar MN, et al (2015): Properties and applications of polyvinyl alcohol, halloysite nanotubes and their nanocomposites. Molecules, 20, 22833–22847.
- 26. Goktepe I, Juneja VK, Ahmedna M (2005): Probiotics in food safety and human health. 1st edn. CRC Press, Boca Raton.
- 27. Han J, Liang C, Cui Y, et al (2018): Encapsulating microorganisms inside electrospun microfibers as a living material enables room-temperature storage of microorganisms. ACS Appl Mater Interfaces, 10, 38799–38806.
- 28. ISO 22117 (2017): ISO. Available at https://www.iso.org/standard/67052.html. (Accessed May 5, 2024).
- 29. ISO/IEC 17043 (2023): ISO. Available at https://www.iso.org/standard/80864.html. (Accessed May 5, 2024).
- 30. ISO 4833-1 (2013): ISO. Available at https://www.iso.org/standard/53728.html. (Accessed May 5, 2024).
- 31. Khan MA, Hussain Z, Ali S, et al (2019): Fabrication of electrospun probiotic functionalized nanocomposite scaffolds for infection control and dermal burn healing in a mice model. ACS Biomater Sci Eng, 5, 6109-6116.
- 32. Kidoaki S, Kwon IK, Matsuda T (2005): Mesoscopic spatial designs of nano- and microfiber meshes for tissue-engineering matrix and scaffold based on newly devised multilayering and mixing electrospinning techniques. Biomaterials, 26, 37–46.
- 33. Koski A, Yim K, Shivkumar S (2004): Effect of molecular weight on fibrous PVA produced by electrospinning. Mater Lett, 58, 493–497.
- 34. Kumar M, Mohania D, Poddar D, et al (2009): A probiotic fermented milk prepared by mixed culture reduces pathogen shedding and alleviates disease signs in rats challenged with pathogens. Int J Probiotics Prebiotics, 4, 211–218.
- 35. Li Q, Jia Z, Yang Y, et al (2008): Preparation and properties of poly (vinyl alcohol) nanofibers by electrospinning. J Polym Eng. 28, 87-100.
- 36. Liu H, Cui SW, Chen M, et al (2019): Protective approaches and mechanisms of microencapsulation to the survival of probiotic bacteria during processing, storage and gastrointestinal digestion: a review. Crit Rev Food Sci Nutr, 59, 2863–2878.
- 37. Mitchell GR, Mohan SD, Davis FJ, et al (2015): Electrospinning: Principles, Practice and Possibilities. The Royal Society of Chemistry, Cambridge.
- 38. Mojaveri SJ, Hosseini SF, Gharsallaoui A (2020): Viability improvement of Bifidobacterium animalis Bb12 by encapsulation in chitosan/poly(vinyl alcohol) hybrid electrospun fiber mats. Carbohydr Polym, 241, 116278.
- 39. Mortazavian AM, Ehsani MR, Mousavi M, et al (2007): Effect of refrigerated storage temperature on the viability of probiotic microorganisms in yogurt. Int J Dairy Technol, 60, 123–127.
- 40. Nagy Z, Wagner I, Suhajda Á, et al (2014): Nanofibrous solid dosage form of living bacteria prepared by electrospinning. Express Polym Lett, 8, 352–361.
- 41. Njjar MS, Akduman Ç, Koluman A (2023): Antibakteriyel, kanama durdurucu ve yaralanma tespit sistemi içeren askeri operasyon kıyafeti. Savunma Bilim Derg, 2, 424–453.
- 42. Ricaurte L, Quintanilla-Carvajal MX (2019): Use of electrospinning technique to produce nanofibres for food industries: a perspective from regulations to characterisations. Trends Food Sci Technol, 85, 92–106.
- 43. Rwei S, Huang C (2012): Electrospinning PVA solution-rheology and morphology analyses. Fibers Polym, 13, 44–50.
- 44. Salalha W, Dror Y, Khalfin RL, et al (2004): Single-walled carbon nanotubes embedded in oriented polymeric nanofibers by electrospinning. Langmuir, 20, 9852–9855.
- 45. Sanz Y (2007): Ecological and functional implications of the acid-adaptation ability of Bifidobacterium: a way of selecting improved probiotic strains. Int Dairy J, 17, 1284–1289.
- 46. Simonič M, Slapničar Š, Trček J, et al (2023): Probiotic Lactobacillus paragasseri K7 nanofiber encapsulation using nozzle-free electrospinning. Appl Biochem Biotechnol, 195, 6768–6789.
- 47. Škrlec K, Zupančič Š, Mihevc SP, et al (2019): Development of electrospun nanofibers that enable high loading and long-term viability of probiotics. Eur J Pharm Biopharm, 136, 108–119.
- 48. Soukoulis C, Singh P, MacNaughtan W, et al (2016): Compositional and physicochemical factors governing the viability of Lactobacillus rhamnosus GG embedded in starch-protein based edible films. Food Hydrocoll, 52, 876–887.
- 49. Tsiouris CG, Tsiouri MG (2017): Human microflora, probiotics and wound healing. Wound Med, 19, 33–38.
- 50. Zavišić G, Ristić S, Petković B, et al (2023): Microbiological quality of probiotic products. Arhiv Za Farmaciju, 73, 17–34.
- 51. Ziabari M, Mottaghitalab V, Haghi AK (2010): A new approach for optimization of electrospun nanofiber formation process. Korean J Chem Eng, 27, 340–354.
There are 51 citations in total.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Microbiology |
| Journal Section | Research Article |
| Authors | |
| Project Number | 2022HZDP011 [ D29 ] |
| Early Pub Date | April 22, 2025 |
| Publication Date | July 1, 2025 |
| Submission Date | July 8, 2024 |
| Acceptance Date | March 20, 2025 |
| Published in Issue | Year 2025Volume: 72 Issue: 3 |
