Innovative screening and application of Lactobacillus paracasei with antioxidant efficacy
308
Presented by: Jinzhao Liu
Abstract
Probiotic Lactobacillus widely exists in human intestine, which is widely used in the food and pharmaceutical field. With the in-depth study of skin microecology, probiotic skincare products have gained extensive attention and application due to their mildness, naturalness and high efficiency. However, most of the probiotic products are embodied in the forms of combinations. and the efficacy claims are confusing, the relevant experimental data are insufficiently supported, as well, the development direction is unclear. In order to make up for the shortcomings of the current research methods in probiotic skincare products, we have established an efficacy-oriented strategy for screening and developing strains with specific antioxidant activity.
2. Methods
With efficacy-oriented screening, the UVB-H2O2 dual-model screening technology was established for preliminary screening, then 4 rounds of screening and verification were carried out by multiple dimensions such as superoxide free radical (ROS) level, the total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity, Catalase (CAT) activity and Nrf2 secretion. Fortunately, a target strain with strong antioxidant effect, Lactobacillus paracasei, was obtained from 85 original probiotics, as well, the Lactobacillus Ferment (LF) product with better stability was prepared.
And then, the safety evaluation of this LF product is carried out by cytotoxicity test with keratinocyte (HaCaT) cells -MTT model, ocular irritation test by Chorioallantoic Membrane Vasular Assay (HET-CAM), as well, human repeated insult patch test (HRIPT) performed on 50 subjects.
In addition, the antioxidant effect of the product was confirmed further by HaCaT-H2O2 damage model, T-AOC ability, SOD & CAT activity, ROS value and Nrf2 secretion level were quantitatively detected by fluorescence spectrophotometer or ELISA determination kit method. Finally, the anti-oxidative and anti-aging effects were tested on the face of 30 healthy subjects.
3. Results
According to the cytotoxicity assay, the viability of HaCaT cells maintain above 80% in the range of 0.5%-10%; with the HET-CAM model, 90% LF is non-irritating, moreover, base on HRIPT test, 10% LF has no potential irritation and induced sensitization, thus LF product, as cosmetic raw material, is safe for skincare.
Based on the HaCaT-H2O2 damage model, compared with the H2O2 damage group, the T-AOC, SOD activity, CAT content and Nrf2 secretion of the experimental group containing 5% LF product were significantly improved by 2274.07%, 483.58%, 3324.24% and 64.83%, respectively; at the same time, the level of ROS was decreased significantly by 37.79% with 5% LF product. From those results, it was found that the Nrf2 antioxidant signal pathway was activated by LF, then antioxidant capacity was enhanced by reduce oxidative damage with T-AOC, SOD and CAT activity. Additionally, in the human efficacy experiment, the water content of the stratum corneum increased by 12% to 14% compared with the placebo within 3 weeks application, the skin elasticity was improved by 10% within one week, and the area of brown spots was reduced by 10% ~12% within three weeks.
4. Discussion and conclusion
Probiotic skincare product with specific efficacy was obtained by efficacy-oriented screening which provided a new ideas and innovative directions. Take Lactobacillus paracasei for example, we focus on the anti-oxidant efficacy and assess the indicators on antioxidant signal pathways, thus the Lactobacillus Ferment (LF) is developed targeting on anti-oxidant and anti-aging. Furthermore, from the in-vivo test results, it is verified the LF product has been shown to firm and brighten skin by increasing skin hydration and skin elasticity, and reducing skin pigmentation.
The above experimental results show that establishing a screening model to obtain strains with specific efficacy can provide a new research method for the development of probiotic skin care products or active materials. At present, the Lactobacillus Ferment product with specific antioxidant effect obtained by this method has a good application prospect in the field of skincare industry.
Probiotic Lactobacillus widely exists in human intestine, which is widely used in the food and pharmaceutical field. With the in-depth study of skin microecology, probiotic skincare products have gained extensive attention and application due to their mildness, naturalness and high efficiency. However, most of the probiotic products are embodied in the forms of combinations. and the efficacy claims are confusing, the relevant experimental data are insufficiently supported, as well, the development direction is unclear. In order to make up for the shortcomings of the current research methods in probiotic skincare products, we have established an efficacy-oriented strategy for screening and developing strains with specific antioxidant activity.
2. Methods
With efficacy-oriented screening, the UVB-H2O2 dual-model screening technology was established for preliminary screening, then 4 rounds of screening and verification were carried out by multiple dimensions such as superoxide free radical (ROS) level, the total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity, Catalase (CAT) activity and Nrf2 secretion. Fortunately, a target strain with strong antioxidant effect, Lactobacillus paracasei, was obtained from 85 original probiotics, as well, the Lactobacillus Ferment (LF) product with better stability was prepared.
And then, the safety evaluation of this LF product is carried out by cytotoxicity test with keratinocyte (HaCaT) cells -MTT model, ocular irritation test by Chorioallantoic Membrane Vasular Assay (HET-CAM), as well, human repeated insult patch test (HRIPT) performed on 50 subjects.
In addition, the antioxidant effect of the product was confirmed further by HaCaT-H2O2 damage model, T-AOC ability, SOD & CAT activity, ROS value and Nrf2 secretion level were quantitatively detected by fluorescence spectrophotometer or ELISA determination kit method. Finally, the anti-oxidative and anti-aging effects were tested on the face of 30 healthy subjects.
3. Results
According to the cytotoxicity assay, the viability of HaCaT cells maintain above 80% in the range of 0.5%-10%; with the HET-CAM model, 90% LF is non-irritating, moreover, base on HRIPT test, 10% LF has no potential irritation and induced sensitization, thus LF product, as cosmetic raw material, is safe for skincare.
Based on the HaCaT-H2O2 damage model, compared with the H2O2 damage group, the T-AOC, SOD activity, CAT content and Nrf2 secretion of the experimental group containing 5% LF product were significantly improved by 2274.07%, 483.58%, 3324.24% and 64.83%, respectively; at the same time, the level of ROS was decreased significantly by 37.79% with 5% LF product. From those results, it was found that the Nrf2 antioxidant signal pathway was activated by LF, then antioxidant capacity was enhanced by reduce oxidative damage with T-AOC, SOD and CAT activity. Additionally, in the human efficacy experiment, the water content of the stratum corneum increased by 12% to 14% compared with the placebo within 3 weeks application, the skin elasticity was improved by 10% within one week, and the area of brown spots was reduced by 10% ~12% within three weeks.
4. Discussion and conclusion
Probiotic skincare product with specific efficacy was obtained by efficacy-oriented screening which provided a new ideas and innovative directions. Take Lactobacillus paracasei for example, we focus on the anti-oxidant efficacy and assess the indicators on antioxidant signal pathways, thus the Lactobacillus Ferment (LF) is developed targeting on anti-oxidant and anti-aging. Furthermore, from the in-vivo test results, it is verified the LF product has been shown to firm and brighten skin by increasing skin hydration and skin elasticity, and reducing skin pigmentation.
The above experimental results show that establishing a screening model to obtain strains with specific efficacy can provide a new research method for the development of probiotic skin care products or active materials. At present, the Lactobacillus Ferment product with specific antioxidant effect obtained by this method has a good application prospect in the field of skincare industry.