Microencapsulation of Probiotic, Evaluation for Viability and Cytotoxic Activities of Its Postbiotic Metabolites on MCF-7 Breast Cancer Cell Line
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 33122
Microencapsulation of Probiotic, Evaluation for Viability and Cytotoxic Activities of Its Postbiotic Metabolites on MCF-7 Breast Cancer Cell Line

Authors: N. V. Enwuru, B. Nkeki, E. A. Adekoya, O. A. Adebesin, B. O. Ojo, R. F. Peters, V. A. Aikhomu, U. E. Mendie, O. Akinloye

Abstract:

Awareness about probiotic health benefits is increasing tremendously. However, cell viability is often low due to harsh conditions exposed during processing, handling, storage, and gastrointestinal transit. Thus, encapsulation is a promising technique that increases cell viability. The study aims to encapsulate the probiotic, evaluate its viability and cytotoxic activity of its postbiotic on the Michigan Cancer Foundation (MCF)-7 breast cancer cell line. Human and animal raw milk was sampled for lactic acid bacteria. Isolated bacteria were identified using conventional and VITEK 2 systems. The identified bacteria were encapsulated using the spray-drying method. The free and encapsulated probiotic cells were exposed to simulated gastric intestinal (SGI) fluid conditions and different storage conditions for their viability. The properties of the formed probiotic granules, their disintegration time, and the weight uniformity of the microcapsules were tested. Furthermore, the postbiotic of the free cells was extracted, and its cytotoxic effect on the MCF-7 breast cancer cell line was tested through [3-(4,5-dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide] (MTT) assay. The bacteria isolated were identified as Lactobacillus plantarum. The size of the formed probiotic granules ranges within 0.71-1.00 mm in diameter, and disintegration time ranges from 2.14 ± 0.045 to 2.91 ± 0.293 minutes, while the average weight is 502.1 mg. The viability of encapsulated cells stored at refrigerated condition (4oC) was higher than that of cells stored at room temperature (25 oC). The encapsulated probiotic cells exhibited better viability after exposure to SGI solution at different pH levels than free cells. The Postbiotic Metabolites (PM) of L. plantarum produced a cytotoxic effect that shows significant activity similar to 5FU, a standard antineoplastic agent. The inhibition concentration of 50% growth (IC50) of postbiotic metabolite was consistent with the IC50 of the positive control (Cisplatin). Lactobacillus plantarum postbiotic exhibited a cytotoxic effect on the MCF-7 breast cancer cell line and could be used as combined adjuvant therapy in breast cancer management. The microencapsulation technique protects the probiotics and maintains their viability.

Keywords: Cytotoxicity effect, encapsulation, postbiotic, probiotic.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 116

References:


[1] H. E Sidjabat and L. Blackall, “One health Probiotics” Microbiology Australia, 2020, https://doi.org/10.1071/ma20016.
[2] C. Ma et al., “Probiotic consumption influences universal adaptive mutations in indigenous human and mouse gut microbiota,” Communications Biology 2021 4:1, vol. 4, no. 1, pp. 1–12, doi: 10.1038/s42003-021-02724-8.
[3] Su, Grace L., Ko, W. Cynthia, Premysl Bercik, Yngve Falck-Ytter Shahnaz Sultan, Weizman V. Adam, and Morgan L. Rebecca, "AGA Clinical Practice Guidelines on the Role of Probiotics in the Management of Gastrointestinal Disorders." Gastroenterology, 2020, https://doi.org/10.1053/j.gastro.2020.05.059.
[4] S. K. Kim et al., “Role of Probiotics in Human Gut Microbiome-Associated Diseases,” J. Microbiol. Biotechnol, vol. 29, no. 9, pp. 1335–1340, Sep. 2019, doi: 10.4014/JMB.1906.06064.
[5] U. Wendel, “Assessing Viability and Stress Tolerance of Probiotics-A Review,” Front Microbiol, vol. 12, Jan. 2022 doi:10.3389/FMICB.2021.818468.
[6] S. Choi, Y. J. Hwang, M. J. Shin, and H. Yi, “Difference in the Gut Microbiome between Ovariectomy-Induced Obesity and Diet-Induced Obesity,” J. Microbiol. Biotechnol., vol. 27, no. 12, pp. 2228–2236, Dec.2017, doi:10.4014/JMB.1710.10001.
[7] G. D’Ettorre et al., “Probiotic supplementation promotes a reduction in T-cell activation, an increase in Th17 frequencies, and a recovery of intestinal epithelium integrity and mitochondrial morphology in ART-treated HIV-1-positive patients,” Immun Inflamm Dis, vol. 5, no. 3, pp.244–260, Sep.2017, doi: 10.1002/IID3.160.
[8] F. Cardoso et al., “Early breast cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow- up†,” Ann Oncol, vol. 30, no. 8, pp.1194–1220, Aug. 2019, doi: 10.1093/ANNONC/MDZ173.
[9] H. Sung et al., “Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries,” CA Cancer J Clin, vol. 71, no. 3, pp. 209–249, May 2021, doi:10.3322/CAAC.21660.
[10] F. Cardoso et al., “European Breast Cancer Conference manifesto on breast centres/units,” Eur J Cancer, vol. 72 pp. 244–250, Feb. 2017, doi:10.1016/J.EJCA.2016.10.023.
[11] S. Chaluvadi, A. T. Hotchkiss, and K. L. Yam, “Gut Microbiota: Impact of Probiotics, Prebiotics, Synbiotics, Pharmabiotics, and Postbiotics on Human Health,” Probiotics, Prebiotics, and Synbiotics: Bioactive Foods in Health Promotion, pp. 515–523, Oct. 2016, doi: 10.1016/B978-0-12-802189-7.00036-8.
[12] M. Sedighi et al., “Therapeutic bacteria to combat cancer; current advances, challenges, and opportunities,” Cancer Med, vol. 8, no. 6, p.3167, Jun. 2019, doi: 10.1002/CAM4.2148.
[13] V. Sankarapandian et al., “An Update on the Effectiveness of Probiotics in the Prevention and Treatment of Cancer,” Life, vol. 12, no. 1, Jan.2022, doi: 10.3390/LIFE12010059.
[14] A. H. Rad, L. A. Maleki, H. S. Kafil, H. F. Zavoshti, and A. Abbasi, “Postbiotics as promising tools for cancer adjuvant therapy,” Adv Pharm Bull, vol. 11, no. 1, pp. 1–5, 2021, doi: 10.34172/APB.2021.007.
[15] Konstantinov SR, Kuipers EJ, Peppelenbosch MP. Functional genomic analyses of the gut microbiota for crc screening. Nat Rev Gastroenterol. 2013; 10:741–5 Foo HL, Loh TC, Law FL, Lim YZ, Kufli CN, Rusul G. Hepatol. Nature publishing group.
[16] J. Angelin and M. Kavitha, “Exopolysaccharides from probiotic bacteria and their health potential,” Int J Biol Macromol, vol. 162, pp. 853–865, Nov. 2020, doi: 10.1016/J.IJBIOMAC.2020.06.190.
[17] H. U. Celebioglu, Y. Erden, and H. B. Ozel, “In vitro cytotoxic effects of lactobacilli grown with lime honey on human breast and colon cancer cells,” Food Biosci, vol. 41, p. 101020, Jun. 2021, doi: 10.1016/J.FBIO.2021.101020.
[18] K. C. Baral, R. Bajracharya, S. H. Lee, and H. K. Han, “Advancements in the Pharmaceutical Applications of Probiotics: Dosage Forms and Formulation Technology,” Int J Nanomedicine, vol. 16, pp. 7535–7556, 2021, doi:10.2147/IJN.S337427.
[19] K. Venema, J. Verhoeven, S. Verbruggen, L. Espinosa, and S. Courau, “Probiotic survival during a multi-layered tablet development as tested in a dynamic, computer-controlled in vitro model of the stomach and small intestine (TIM-1),” Lett Appl Microbiol, vol. 69, no. 5, pp. 325–332, Nov. 2019, doi: 10.1111/LAM.13211.
[20] D. de A. Paula, E. M. F. Martins, N. de A. Costa, P. M. de Oliveira, E.B de Oliveira, and A. M. Ramos, “Use of gelatin and gum arabic for microencapsulation of probiotic cells from Lactobacillus plantarum by a dual process combining double emulsification followed by complex coacervation,” Int J Biol Macromol, vol. 133, pp. 722–731, Jul. 2019, doi: 10.1016/J.IJBIOMAC.2019.04.110.
[21] S. Razavi, S. Janfaza, N. Tasnim, D. L. Gibson, and M. Hoorfar, “Microencapsulating polymers for probiotics delivery systems: Preparation, characterization, and applications,” Food Hydrocoll, vol.120, p. 106882, Nov.2021, doi: 10.1016/J.FOODHYD.2021.106882.
[22] H. Liu et al., “Protective approaches and mechanisms of microencapsulation to the survival of probiotic bacteria during processing, storage a gastrointestinal digestion: A review,” Crit Rev Food Sci Nutr, vol. 59, no. 17, pp.2863–2878, Sep. 2019, doi:10.1080/10408398.2017.1377684.
[23] N. Choudhury, M. Meghwal, and K. Das, “Microencapsulation: An overview on concepts, methods, properties and applications in foods,” Food Front, vol. 2, no. 4, pp. 426–442, Dec. 2021, doi:10.1002/FFT2.94.
[24] L. Huan, W. C. Steve, C. Maoshen, I. Yue, L. Rong, X. Feifei & Z.Fang (2019) Protective approaches and mechanisms of Microencapsulation to the survival of probiotic bacteria during processing, storage and gastrointestinal digestion: areview; Critical Reviews in Food Science and Nutrition, https://doi.org/10.1080/10408398.2017.1377684.
[25] Y. Taye, T. Degu, H. Fesseha, and M. Mathewos, “Isolation and Identification of Lactic Acid Bacteria from Cow Milk and Milk Products,” Scientific World Journal, vol. 2021, 2021, doi:10.1155/2021/4697445.
[26] Y. B. Soemarie, T. Milanda, and M. I. Barliana, “Isolation, Characterization, and Identification Candidate of Probiotic Bacteria Isolated from Wadi Papuyu (Anabas testudineus Bloch.) a Fermented Fish Product from Central Kalimantan, Indonesia,” Int J Food Sci, vol.2022, 2022, doi:10.1155/2022/4241531.
[27] L. O. Chuah et al., “Postbiotic metabolites produced by Lactobacillus plantarum strains exert selective cytotoxicity effects on cancer cells,” BMC Complement Altern Med, vol. 19, no. 1, pp. 1–12, Jun. 2019, doi:10.1186/S12906-019-2528-2/TABLES/3.
[28] D. N. Pujara and B. R. Parmar, “Formulation and Evaluation of Hard Gelatin Capsule of Losartan Potassium,” Inventi Rapid: Pharm Tech, vol. 2013, no. 2, pp. 1–4, 2013, (Online). Available: www.inventi.in.
[29] B. D. Kumar, and N. Sharma. "Design, Formulation and Evaluation of Fast Disintegrating Tablets of Glipizide."2019. https://doi.org/10.22270/jddt.v9i3-s.2806
[30] P. Hrishav Das, and B. Nath. "Formulation and Evaluation of Oral Fast Disintegrating Tablet of Ibuprofen Using Two Super Disintegrants." International Journal of Current Pharmaceutical Research; Vol 9, Issue 4, 2017, https://core.ac.uk/download/482244375.
[31] United States Pharmacopeia and National Formulary (USP 36- NF 31), “Disintegration 701,” in United States Pharmacopeia Convention, Rockville, MD, 2013, pp. 305–307.
[32] D. Arepally, R. S. Reddy, and T. K. Goswami, “Studies on survivability, storage stability of encapsulated spray dried probiotic powder,” Curr Res Food Sci, vol. 3, pp. 235–242, Nov. 2020, doi 10.1016/J.CRFS.2020.09.001.
[33] H. K. Tan, H. L. Foo, T. C. Loh, N. B. M. Alitheen, and R. A. Rahim,“Cytotoxic effect of proteinaceous postbiotic metabolites produced by Lactobacillus plantarum I-UL4 cultivated in different media composition on MCF-7 breast cancer cell,” Malays J Microbiol, vol. 11, no. 2, pp.207–214, Jun. 2015, Accessed: Oct. 12, 2023. (Online) Available: https://doaj.org/article/fc6f9a748a19405790ab20aae5179e2e.
[34] L. K. Ursell, J. L. Metcalf, L. W. Parfrey, and R. Knight, “Defining the human microbiome,” Nutr Rev, vol. 70, no. 1, pp. 38–44, Aug. 2012, doi: 10.1111/J.1753-4887.2012.00493. X.
[35] Stasiak-Róza´nska, L.; Berthold-Pluta, A.; Pluta, A.S.; Dasiewicz, K.; Garbowska, M. Effect of Simulated Gastrointestinal Tract Conditions on Survivability of Probiotic Bacteria Present in Commercial Preparations. Int. J. Environ. Res. Public Health 2021, 18, 1108; https://doi.org/10.3390/ijerph18031108
[36] Hoetelmans RM. "IC50 versus EC50". PK-PD relationships for anti-retroviral drugs. Amsterdam: Slotervaart Hospital. Archived from the original on 2017-05-28 – via U.S. Food and Drug Administration.
[37] S. Huang, F. Gaucher, C. Cauty, J. Jardin, Y. L. Loir, R. Jeantet et al., Growth in hyper-concentrated sweet whey triggers multing, stress tolerance and spray drying survival in Lactobacillus casei BL23: from the molecular basis to new perspectives for sustainable probiotic production. Front. Microbiol.9:2548, doi: 10.3389/fmicb.2018.02548.
[38] N. Kabilan, M. Murugesan, T. Balasubramanian, S. Geethalakshmi. “Physico-Chemical Analysis of Siddha Drug Poora parpam - A Comparative Evaluation between Natural and Synthetic Source”, Scholars Research Library, European Journal of Applied Engineering and Scientific Research, 2017, 5(2):6-14.
[39] A. Punitha, S. Visweswaran, N.J. Muthukumar, “Physico-chemical evaluation and hptlc finger print of siddha poly herbal formulation “Swasa kudori mathirai” 2015.https://core.ac.uk/download/482239090.pdf.
[40] Bulk Density and Tapped Density of Powders; Stage 6 Harmonization Official August 1, 2015. https://www.usp.org/.../gen-chapter/bulk_density
[41] M.A. Kaleem, M. Z. Alam, M. Khan, M., Jaffery, S. H. I., & Rashid, B. An experimental investigation accuracy of Hausner Ratio and Carr Index of powders in additive manufacturing processes. Metal Powder Report. doi:10.1016/j. mprp.2020.06.061.
[42] United States Pharmacopeia National Formulary (USP 36- NF 31), “Disintegration 701,” in United States Pharmacopeia Convention, Rockville MD, 2013, pp. 305–307.
[43] A. M. Agiba, S. S. Abul-Ella, Rehab A. Abd El-Monem, “Pharmaco-technical Development and Optimization of Multilayered Tablets: An Updated Industrial Review with Emphasis on Bilayer Tablets”, International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 13, Issue 4, 2021. DOI: https://dx.doi.org/10.22159/ijap.2021v13i4.41528.
[44] U. Wendel, “Assessing Viability and Stress Tolerance of Probiotics”, Front. Microbio., Vol.12- 2022; https://doi.org/org/10.3389/fmicb.2021.818468
[45] W. Fiore, S. Arioli and S. Guglielmetti, The neglected microbial components of commercial probiotic formulations. Microorganisms 2020, doi:10.3390/microorganisms8081177
[46] S. Han, Y. Lu, J. Xie, Y. Fei, G. Zheng, Z. Wang, J. Liu, L. Lv, Z. Ling, B. Berglund, M. Yao and L. Li (2021) Probiotic Gastrointestinal Transit and Colonization After Oral Administration: A Long Journey; Front. Cell. Infect. Microbiol. 11:609722. doi:10.3389/fcimb.2021.609722.