A novel Cordyceps Militaris Ferment Extract with Chinese characteristics targets on well aging
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Presented by: Shaojing Sun
1. Introduction:
Cordyceps militaris, a traditional herbal drugs in China, has gained considerable significance in several clinical and biotechnological applications, while it is difficult to be fully extracted the bioactive constituents from C. militaris, by traditional extraction methods. Generally, bio-fermentation is one innovative technology to extract active substances effectively, especially, probiotics and fermentation by probiotics is famous in food, cosmeitics and pharmaceuticals industry.
In order to gain the active substances, we developed the Cordyceps Militaris Ferment Extract (CMF) through fermentation with probiotics as strain, and it have been proved that the active components were increased significantly than common extraction method. Moroever, the potential applications of CMF in cosmetics and medicine industry have been carried out by in vitro and in-vivo studies.
2. Methods used:
First of all, the Cordyceps Militaris Ferment Extract (CMF) was obtained by high-efficiency fermentation process using highly active Lactobacillus as the strain and Cordyceps militaris as the main substrate, and the main active substances were quantitatively detected, such as cordycepin by HPLC method, cordycepic acid through colorimetric method. Then the cell cytotoxicity was assessed by the WST method with Human keratinocytes (HaCaT) cells; the safety evaluation in human skin was evaluated by the single closed skin patch test. Further, the multi-dimensional anti-ageing tests have being carried out, for anti-oxidant efficacy, the scavenging rate of hydroxyl free radicals (·OH) and superoxide free radical (ROS) were measured by chemical method and HaCaT-UV damage model (in-vitro test) respectively; for anti-inflammatory effect, the expression value of inflammatory factors were monitored in mice macrophages model (in-vitro test); moreover, for the well aging, the expression level of collagen I was measured with fibroblast model using immunofluorescence statistics; at last, the moisturizing and anti-aging efficacy were researched on the skin of 30 healthy volunteers.
3. Results obtained:
The content of main active substances were improved significantly after lactobacillus fermentation, the value of cordycepin was up to 180 ppm, as well, cordycepic acid was increased to 2%. According to the WST assay and patch test, the viability of HaCaT cells don’t show any dramatic change and maintain above 80% in the range of ≤5% for CMF, moreover, it was proved that 2% CMF, as cosmetics active ingredient, was safe in skin.
In anti-oxidant test, it was found the 67.8% of ·OH could be scavenged with 2% CMF; and the scavenging rate of ROS was as high as 35.7% at 1.5%.CMF. For anti-inflammatory research, the inhibition ratio of IL-1β, IL-6 and TNF-β were 100%, 11.5% and 8.7%. Furthermore, the expression level of collagen I was up-regulated by 90% with 0.01% CMF. Finally, several in-vivo tests had been carried out, the results show CMF has a better water-binding ability, moreover, CMF significantly improve the skin elasticity after continuous application of a facial cream containing 1% CMF.
All the results indicate that CMF has a vast potential application and effectiveness in medicine and cosmetics industry.
4. Discussion and conclusion:
Fermentation by probiotics is an effective manner of active ingredient extraction and enrichment, after fermentation, the value of active ingredients in CMF were greatly increased, especially cordycepin and cordycepic acid, which is the foundation of skin protective effect.
Our findings conclude that CMF at lower concentration effectively, allround ameliorated the skin aging caused by oxidant damage, inflammatory response and intrinsic aging, thus CMF may consider as a desirable cosmetics ingredient for skin protection and/or preparation of skin care products.
Cordyceps militaris, a traditional herbal drugs in China, has gained considerable significance in several clinical and biotechnological applications, while it is difficult to be fully extracted the bioactive constituents from C. militaris, by traditional extraction methods. Generally, bio-fermentation is one innovative technology to extract active substances effectively, especially, probiotics and fermentation by probiotics is famous in food, cosmeitics and pharmaceuticals industry.
In order to gain the active substances, we developed the Cordyceps Militaris Ferment Extract (CMF) through fermentation with probiotics as strain, and it have been proved that the active components were increased significantly than common extraction method. Moroever, the potential applications of CMF in cosmetics and medicine industry have been carried out by in vitro and in-vivo studies.
2. Methods used:
First of all, the Cordyceps Militaris Ferment Extract (CMF) was obtained by high-efficiency fermentation process using highly active Lactobacillus as the strain and Cordyceps militaris as the main substrate, and the main active substances were quantitatively detected, such as cordycepin by HPLC method, cordycepic acid through colorimetric method. Then the cell cytotoxicity was assessed by the WST method with Human keratinocytes (HaCaT) cells; the safety evaluation in human skin was evaluated by the single closed skin patch test. Further, the multi-dimensional anti-ageing tests have being carried out, for anti-oxidant efficacy, the scavenging rate of hydroxyl free radicals (·OH) and superoxide free radical (ROS) were measured by chemical method and HaCaT-UV damage model (in-vitro test) respectively; for anti-inflammatory effect, the expression value of inflammatory factors were monitored in mice macrophages model (in-vitro test); moreover, for the well aging, the expression level of collagen I was measured with fibroblast model using immunofluorescence statistics; at last, the moisturizing and anti-aging efficacy were researched on the skin of 30 healthy volunteers.
3. Results obtained:
The content of main active substances were improved significantly after lactobacillus fermentation, the value of cordycepin was up to 180 ppm, as well, cordycepic acid was increased to 2%. According to the WST assay and patch test, the viability of HaCaT cells don’t show any dramatic change and maintain above 80% in the range of ≤5% for CMF, moreover, it was proved that 2% CMF, as cosmetics active ingredient, was safe in skin.
In anti-oxidant test, it was found the 67.8% of ·OH could be scavenged with 2% CMF; and the scavenging rate of ROS was as high as 35.7% at 1.5%.CMF. For anti-inflammatory research, the inhibition ratio of IL-1β, IL-6 and TNF-β were 100%, 11.5% and 8.7%. Furthermore, the expression level of collagen I was up-regulated by 90% with 0.01% CMF. Finally, several in-vivo tests had been carried out, the results show CMF has a better water-binding ability, moreover, CMF significantly improve the skin elasticity after continuous application of a facial cream containing 1% CMF.
All the results indicate that CMF has a vast potential application and effectiveness in medicine and cosmetics industry.
4. Discussion and conclusion:
Fermentation by probiotics is an effective manner of active ingredient extraction and enrichment, after fermentation, the value of active ingredients in CMF were greatly increased, especially cordycepin and cordycepic acid, which is the foundation of skin protective effect.
Our findings conclude that CMF at lower concentration effectively, allround ameliorated the skin aging caused by oxidant damage, inflammatory response and intrinsic aging, thus CMF may consider as a desirable cosmetics ingredient for skin protection and/or preparation of skin care products.