Title : Microencapsulation of lactobacillus plantarum probiotic and evaluation for viability, antimicrobial property and cytotoxic activities of its postbiotic metabolites on mcf-7 breast cancer cell line.
Abstract:
Background: Probiotics are live microbial feed supplement that beneficially affects the host. Several probiotics as well as their postbiotic products have been used for prevention or treatment of various conditions. However, cell viability in these products is often low due to harsh conditions that come across during processing, handling, storage, and gastrointestinal conditions. These strongly influence the benefits that probiotics can provide, thus, viability is an important parameter for probiotic to produce health benefits on the host. Microencapsulation is a promising technique with considerable effects on probiotic survival. The study is aimed to formulate microencapsulated probiotic and evaluate its viability, antimicrobial efficacy and cytotoxic activity of its postbiotic on MCF-7 breast cancer cell line.
Method: Human breast milk, infant feaces and breast milk from animals (cows, goats and sheep) were sampled for lactic acid bacteria, and the isolated bacteria were identified using conventional and vitek 2 system. The identified lactic acid bacterium was encapsulated using spray dried and extrusion methods. The free, encapsulated and chitosan coated encapsulated probiotics were tested for viability in simulated-gastric intestinal (SGI) fluid and in different storage conditions, at refrigerated (4 oC) and room (25oC) temperatures. The disintegration time and weight uniformity of the spray dried hard gelatin capsules were tested. Furthermore, antimicrobial property of free and encapsulated probiotic was tested against enteric pathogenic isolates from antiretroviral therapy (ART) treated HIV positive patients. The postbiotic of the free cells was extracted and its cytotoxic effects on MCF-7 breast cancer cell line was tested through MTT assay.
Result: Lactic acid bacterium was isolated from cow breast milk and was identified as Lactobacillus plantarum. It was designated Lactobacillus plantarum NBL1. A zero-size hard gelatin Lactobacillus plantarum NBL1 capsules with granules within a size range of 0.71–1.00 mm diameter was formulated. The disintegration time ranges from 2.14±0.045 to 2.91±0.293 minutes, while the average weight is 502.1mg. The viability of free cells stored at 4o C was less than 3 log CFU/g after 6months, while encapsulated cells maintain the highest viability, 8 log CFU/G at 4o C but lost little viability, less than 6 log CFU/g at 25o C. In simulated gastrointestinal juice, encapsulated cells were released, viable and effective against the enteric pathogens tested. However, chitosan coated calcium alginate encapsulated probiotic significantly inhibited Shigella flexneri, Candida albicans and Escherichia coli. The Postbiotic Metabolites (PM) of L. plantarum produced cytotoxic effect on MCF-7 breast cancer cell line. The postbiotic showed significant cytotoxic activity very similar to 5FU, a standard antineoplastic agent. The inhibition concentration of 50% growth (IC50) of postbiotic metabolite, K3 is low and consistent with the IC50 of the positive control (Cisplatin).
Conclusions: Lactobacillus plantarum postbiotic exhibited cytotoxic effect on MCF-7 breast cancer cell line and could be used as combine adjuvant therapy in breast cancer management. Microencapsulation technique provides protection to probiotic, improves its viability and delivery to gastrointestinal tract. Chitosan enhances antibacterial efficacy, thus, chitosan coated microencapsulated L. plantarum probiotic could be more effective and use as combined therapy in HIV management of opportunistic enteric infection.
Audience Take Away:
- Probiotic is a good supplement that boost good health.
- Microencapsulation technique provides protection to probiotic, improves its viability and delivery to gastrointestinal tract.
- Thus, microencapsulation will provide solution to survival and viability of probiotic in some food matrices; since the analysis of probiotic products in several different countries has confirmed that probiotic strains exhibit poor survival in food such as fermented dairy products (Shah, 2000).
- Postbiotic product of L. plantarum could be used as combine adjuvant therapy in breast cancer management. Studies have shown that clinical health merits of LAB metabolites associated with the cancer therapies include the establishment of eubiosis conditions in the gut ecosystem, contribution to the recovery (after cancer surgery) and decreasing hospitalization period, eliminating superficial incisional surgical position infection, and preventing some side effects of conventional cancer therapies (chemotherapy and antibiotic-induced diarrhea) (Aisu et al., 2015; Tan et al., 2016 and Homayouni et al., 2020).