<p>The Efficacy of Electrospun PAN/Kefiran Nanofiber and Kefir in Mammalian Cell Culture: Promotion of PC12 Cell Growth, Anti-MCF7 Breast Cancer Cells Activities, and Cytokine Production of PBMC</p>

Abstract

Background: Kefiran is a useful polysaccharide made of branched glucogalactose which is produced by microorganisms. Here the anti-MCF-7 breast cancer cells activity of kefiran and cytokine productions (IL-6) of peripheral blood mononuclear cells (PBMC) treated by kefiran was studied. Also, the effect of using kefiran as a useful and cost-effective scaffold in neural stem cell culture (PC12 cell culture) was investigated. Material and Methods: Kefiran was produced from raw milk with 0.5% fat and 10 g of kefir grains. After incubation for 48 hrs at room temperature, the solvent collected (crude kefiran). These samples were kept at 100°C for 1 hr (boiled kefiran) and the supernatant was precipitated by ethanol (pure kefiran). Then, the electrospun nanofibers, pure polyacrylonitrile (PAN), PAN/kefiran 5%, and PAN/kefiran 10% were fabricated and used as scaffolds in the cell culture. The structure of fabricated was studied by SEM and the cytokine production (IL-6) in vitro in the cell culture supernatant of PBMC line after treatment with kefiran (1mg/mL, 5 mg/mL) and kefiran-PAN 5% and 10% were carried out by enzyme-linked immunosorbent assay (ELISA). The attachment of PC12 cells was examined by inverted microscope. Also, cytotoxicity of kefiran for PC12 and MCF7 cells and morphological changes of PC12 cells were evaluated by MTT and Cresyl violet staining (Nissl staining) respectively. Results: The mean diameter of fabricated PAN/kefiran 5% and 10% nanofibers were 310.2±43.97 nm. The contact angle measurement results (26.9± 1.9 for the pure PAN scaffold vs 12.3± 1.13 for the PAN/kefiran) revealed enhanced hydrophilicity of scaffolds upon the incorporation of kefiran and PAN. Seeding of PC12 cells on the scaffolds showed that fabrication of kefiran into PAN led to the enhancement of cell attachment, proliferation, and morphological changes. Also, the promotion of PBMC growth and decreasing of MCF7 cell lines viability were shown through MTT assay. No significant changes were measured for the level of IL-6 in PAN/kefiran 5% treated cells compared to the control (p ≥ 0.05). Conclusion: These results suggest superior properties of kefiran/PAN nanofibrous scaffolds for the neural stem cell culture especially for repairing injured spinal cord. Also, the pure kefiran could be used for the enhancement of PBMC growth and reducing the MCF7 cancerous cells growth. So, using biocompatible, anti-bacterial, and anti-tumor kefiran/ PAN nanofibers for regenerative medicine seems promising.

Bioactive Compounds from Kefir and Their Potential Benefits on Health: A Systematic Review and Meta‐Analysis

“…For EPS, although the intervention time has been similar for both a pro- and anti-inflammatory effect assay, the EPS concentration employed was dramatically higher; in vitro , the concentration for immunostimulating varied from 50 to 5000 μ g/mL [ 4 , 46 , 51 ], while for inhibition, it reached the maximum of 100 μ g/mL [ 50 ]. In vivo , 100 mg/kg bw was administered orally for immunostimulating [ 58 ], while to inhibit the immune response, the concentration ranged from 0.03 to 3 mg/kg bw [ 50 ].…”

“…EPSs were as efficient as lipopolysaccharides in promoting cell proliferation, phagocytosis, and cytokine secretion by macrophages [ 51 , 52 ]. Kefiran at 1000 to 5000 μ g/mL increased IL-6 secretion, and concentrations from 2000 to 4000 μ g/mL stimulated cell proliferation of human PBMC culture by up to 200% after 24 h [ 46 ]. Bioactive peptide from L. kefiranofaciens , turn on, improved secretion of TNF α , IL-1 β , IL-6, and IL-12 by 1000, 700, 1300, and 3000% by murine peritoneal macrophage culture.…”

“…Kefiran reduced up to 45% of MCF7 breast cancer cell viability after 48 h of intervention, losing effect under the highest tested concentration of 4 mg/mL. However, efficiency decreased to 15.6% after 72 h, without effect at the lowest tested concentration of 0.5 mg/mL [ 46 ]. For HepG2 hepatocellular carcinoma cells, kefiran reduced their viability by up to 82% from the minimum concentration of 250 μ g/mL.…”

“…Finally, the HT-29 colon cancer cell line [ 47 , 49 ] appeared to be more sensitive to EPS than the MCF7 breast cancer cell line [ 46 ]. For the latter, the concentration reported for the assay was ten times greater ( Table 2 ).…”

In-Vitro Cytotoxicity Study: Cell Viability and Cell Morphology of Carbon Nanofibrous Scaffold/Hydroxyapatite Nanocomposites

“…To regulate the actions of cells, tissue engineering (TE) uses physical, chemical, biological, and engineering processes. More specifically, the field of regenerative medicine (RM) is defined as the application of TE and is related to the principles of biology and engineering that restore the structure and function of damaged tissues and organs [ 1 ]. This involves not only in vivo, but also in vitro functional tissue generation ideal for various purposes, such as drug testing, disease models, including cell/tissue/organ one-chip approaches [ 2 ].…”