Phytocelltec™ Goji Stem Cell Activation for a V-Shaped Face

PhytoCellTec™ Goji Stem cell activation for a V-shaped face

Goji Plant Stem Cells Vitalize Skin Stem PhytoCellTec™ Goji Cells for an Improved Face Shape • Stimulates exosome signaling PhytoCellTec™ Goji is based on a plant stem cell • Increases collagen and elastin expression culture of a goji seedling that vitalizes mesenchymal • Improves skin density stem cells to ensure an improved V-shaped face. • Tightens facial contours • Minimizes fine wrinkles and deeper lines Goji (Lycium barbarum) plants are native to Southeast Europe and Asia. The red-orange goji berries that are Applications harvested from this plant are one of the most famous • Tightening and contouring serums for the face and neck superfruits. And thanks to our PhytoCellTec™ tech • Collagen boosting formulations nology, we are able to produce goji plant stem cells • Contouring masks that can be used in cosmetic formulations in order to • Lifting and firming anti-aging formulas improve the function of skin stem cells. Formulating with PhytoCellTec™ Goji As we age, the production of collagen and elastin is • Recommended use level: 0.4 – 1 % reduced, which results in sagging skin that can most • Incorporation: For cold processes, dissolve notably be observed at the face contours of the jawline. PhytoCellTec™ Goji into the aqueous phase. In hot / cold processes, add during the cooling Mesenchymal stem cells have been shown to improve phase below 40 °C. collagen production and regenerate the skin, for example • Thermostability: Temperatures of up to 40 °C during wound healing. These processes are mediated for a short time will not affect the stability of by vesicles known as exosomes which are produced and PhytoCellTec™ Goji. secreted by these stem cells. INCI (EU / PCPC) Declaration PhytoCellTec™ Goji was shown to: Lycium Barbarum Callus Culture Extract (and) • improve the stemness of aged mesenchymal stem cells Isomalt (and) Lecithin (and) Aqua / Water • increase exosome signaling by mesenchymal stem cells, which in turn improves extracellular matrix production Additional Information in fibroblasts. • Not preserved • Alcohol-free The improved extracellular matrix rejuvenates the skin by improving skin density, reducing wrinkles and reshaping the face for an improved oval face shape.

October 2019 PhytoCellTec™ Goji Goji plant stem cells for skin stem cells

Goji – the superfruit PhytoCellTec™ by Mibelle Biochemistry Goji (Lycium barbarum) plants are native to Southeast Mibelle Biochemistry developed the PhytoCellTec™ Europe and Asia. They belong to the nightshade family technology to enable the large scale cultivation of – just like tomatoes and potatoes – and they grow up plant stem cells. This technology relies on the wound to 3 meters high. The red-orange goji berries that are healing mechanism of a plant, whereupon part of a harvested from this plant are one of the most famous plant is wounded in order to induce the formation of superfruits. callus cells. This healing tissue consists of dedifferen tiated cells which are stem cells. Callus cells are har- According to legend, goji berries were first discovered by vested and cultivated in a suspension in an innovative a Buddhist monk and it transpired that those monks bioreactor. who incorporated them into their diets lived longer than those who did not. For hundreds of years, goji berries PhytoCellTec™ Goji is based on a plant stem cell have been eaten in Asia for their medicinal and nutritive culture of a goji seedling. By producing a plant properties. Nowadays, goji berries are eaten all over stem cell culture of goji for cosmetics, the optimal the world, mostly in dried form, and there are various goji composition to improve skin stem cell function can berry supplements on the market with the reported be achieved. effect claims ranging from anti-aging, immune boosting and energizing to improved memory. Exosomes Dermal rejuvenation for a V-shaped face

Mesenchymal Stem Cells – of Importance for Improved V-Shape Via Dermal Rejuvenation Regenerative Therapeutics When our skin ages, the vitality of fibroblasts is reduced Mesenchymal stem cells are precursors of different cell and this results in reduced extracellular matrix production types, including dermal skin cells, and they were shown and sagging skin. In addition, mesenchymal stem cells are to be essential for various regeneration processes, such also aging and this reduces their regenerative properties. as wound healing. Interestingly, it has been demon- strated that predominantly mesenchymal stem cells are PhytoCellTec™ Goji revitalizes mesenchymal stem cells not needed for this process, but rather vesicles known and boosts their exosome production. This results in the as exosomes that are secreted by these cells. improved production of extracellular matrix components such as collagen and elastin by fibroblasts. Cell-to-Cell Communication via Exosomes Therefore, one can support the dermal rejuvenation of Exosomes are small, approximately 100 nm vesicles that the skin by stimulating mesenchymal stem cells in order are surrounded by a membrane which contains messenger to produce more exosomes. This will in turn lead to a molecules such as RNAs, proteins and lipids. Exosomes reduced wrinkle appearance, less sagging and thus an derived from mesenchymal stem cells have been shown improved V-shaped face. to have a beneficial effect on other cell types such as fibroblasts and are currently being heavily investigated for their diagnostic and therapeutic purposes. The topic of exosomes is quite new; while in 2006 there were less than 100 scientific articles on this subject, there were more than 9,000 scientific articles in 2018. This underlines the unquestionable relevance of this relatively new field.

Exosomes from Mesenchymal Stem Cells Rejuvenate Fibroblasts

Collagen

Fibroblast Fibroblast

PhytoCellTec™ Goji Fusion of exosome with the ﬁbroblast. Activation of collagen Exosome expression.

Mesenchymal stem cell Mesenchymal stem cell PhytoCellTec™ Goji Study results

Improving the Stemness of Aged Mesenchymal Stem cells To investigate whether PhytoCellTec™ Goji influences the vitality of mesenchymal stem cells (MSCs), adipose-derived human MSCs were grown for 14 pas- sages to mimic the aging process. These aged MSCs were then either treated or not (control) with 1 % Goji stem cell extract for 72 hours. The expression of stem cell markers was then assessed with RT-qPCR.

Results showed that treatment with Goji stem cell extract increased the expression of the stem cell markers CD34, Oct4 and Nanog in aged MSCs. This means that PhytoCellTec™ Goji is able to rejuvenate mesen chymal stem cells and help them maintain their stemness.

Increase in Stem Cell Marker Expression

Control 1 % Goji stem cell extract

8 ol (=1) 6

4 malized to contr

nor 2 Fold change of stem cell markers

0 Control CD34 Oct4 Nanog

S-707 / © Mibelle Biochemistry Increase in Exosome Production Stimulation of Extracellular Matrix Genes by Mesenchymal Stem Cells through Cell-to-Cell Communication In order to assess the effect of PhytoCellTec™ After demonstrating the effect on exosome Goji on the exosome production capability, human mes- production in MSCs, it was investigated as to whether enchymal stem cells (MSCs) were either treated or not mesenchymal stem cells treated with Goji stem cell (control) with 0.1 % Goji stem cell extract for a period of extract are able to communicate with fibroblasts in order 24 hours. The exosomes that were released from the to produce more extracellular matrix (ECM) proteins. cells were isolated and quantified in two different ways: quantification of total protein amount and quantification For this purpose, human MSCs were treated with 1 % of the activity of Acetylcholinesterase, which is a known Goji stem cell extract for a period of 72 hours. The super- exosomal protein. natant, which includes soluble factors as well as exosomes («conditioned medium»), was added to fibroblasts for a Both quantification methods revealed that treatment period of 24 hours, which was followed by gene expres- with Goji stem cell extract leads to an increase in exo- sion analysis by RT-qPCR. Fibroblasts treated with medium some production by MSCs. from untreated MSCs served as the control.

Results showed that the conditioned medium from Goji stem cell extract-treated MSCs increased the gene expression of several ECM factors such as collagen 3 and 16 and Elastin. PLOD1 is crucial for collagen production, while LOX (lysyl oxidase) connects collagen and elastin to ensure more stability and elasticity. The direct treatment of the fibroblasts did not have the same effect. Therefore, PhytoCellTec™ Goji improves cell-to-cell communication between MSCs and fibroblasts.

Increase in Exosome Production Improved Cell-to-Cell Communication

Control 0.1 % Goji stem cell extract Conditioned medium from untreated MSCs Conditioned medium from MSCs treated with 1 % Goji stem cell extract 2 300

1.5 *

ession 200 ol (=1) ol (=100) in % 1

100 ed to contr malized to contr ease in exosome levels 0.5 nor Incr Change in gene expr compar

0 0 Control COL3A1 COL16A1 Elastin PLOD1 LOX Total protein Enzyme activity quantiﬁcation quantiﬁcation

*p<0.05 versus control

S-762 / © Mibelle Biochemistry S-799/ © Mibelle Biochemistry PhytoCellTec™ Goji Study results

Improvement of Wrinkle Depth Twenty-three Caucasian women aged between In addition, pictures of the face were taken with 41 and 69 years (mean age: 56 years) who dis- Visioscan. played signs of photo-aging applied either a cream with 0.4 % PhytoCellTec™ Goji on one-half of the face and As shown in the pictures below, treatment with 0.4 % the corresponding placebo on the other half of the face, PhytoCellTec™ Goji improved the appearance of twice daily for a period of 56 days. The wrinkle depth wrinkles in the eye area. Notably, not only fine wrinkles was measured using PRIMOS lite. below the eye appeared smoother (see upper panel), but deeper lines were also visibly improved (see lower Treatment with PhytoCellTec™ Goji led to an average panel). decrease in wrinkle depth of 11 % compared to the placebo.

Decrease in Wrinkle Depth on Crow’s Feet Visible Smoothing of the Eye Area

Placebo 0.4 % PhytoCellTec™ Goji Before After 56 days 14

12 *

8 Before After 56 days 6 ease in wrinkle depth

ed to initial conditions in % 4 Decr

compar 2

*p<0.05 versus initial conditions

S-755 / © Mibelle Biochemistry S-755 / © Mibelle Biochemistry Improvement of Skin Density in Photo-Aged Skin Twenty-three Caucasian women aged between image. However, disruption to this regular architecture 41 and 69 years (mean age: 56 years) who displayed leads to weaker reflections and dark patches, as can signs of photo-aging applied a cream with 0.4 % Phyto- be seen in the ultrasonographic images before at Day 0. CellTec™ Goji on the inner side of one forearm and These so-called subepidermal low-echogenic bands the corresponding placebo cream on the inner side of (SLEB) are commonly found in aged and photo-dam- the other forearm twice daily for a period of 28 days. aged skin. The density of their skin (epidermis + dermis) was determined by ultrasonic measurements. Treatment with 0.4 % PhytoCellTec™ Goji for one month significantly increased skin density by 6 % compared to The collagen and elastic fiber structure of an intact initial conditions and reduced the SLEB in photo-aged dermis yields many reflections from the ultrasonic wave skin. that are visible as bright colors in the ultrasonographic

Increase in Skin Density

Placebo 0.4 % PhytoCellTec™ Goji Before After 28 days 8

ed to 6 **

2 eated and initial conditions in % ease in skin density compar untr Incr

*p<0.05 versus untreated

mis mis **p<0.01 versus untreated SLEB Der Epider

Improvement of Oval Face Shape Sixty-seven Caucasian women aged between the face, vertical parallel lines were drawn to intersect at 39 and 70 years (average age 57 years) with the start of the jawline. The points of the jawline inter- sagging facial skin were split into two groups. One group section were then connected with the middle of the chin applied a cream with 0.4 % PhytoCellTec™ Goji and (see illustration below). The area below the drawn V lines the other group applied the corresponding placebo cream was determined. A reduction of this area would suggest on the entire face and neck twice daily for 28 days. a reduction in facial sagging and an improved V-shape of the face. The facial sagging and oval face shape was determined by measuring the size of the neck / submandibular triangle area by image analysis of the pictures of the volunteers’ faces that were taken with Visioface.

For the image analysis, a straight line was drawn verti- cally through the middle of the chin. On either side of

Measurement of the V-Shape of the Face

PhytoCellTec™ Goji Results showed that treatment for four weeks with 0.4 % PhytoCellTec™ Goji significantly improved the oval face shape by 6 % compared to initial conditions.

The improved V-shape of the face was also visible in the Visioface pictures of the jawline.

This demonstrates that PhytoCellTec™ Goji is indeed able to reduce facial sagging and improve the oval shape of the face.

Improved V-Shape of the Face

Placebo 0.4 % PhytoCellTec™ Goji

8 ea

6 * Before

4 ed to initial conditions in % 2

compar After ease in neck / submandibular triangle ar 28 days

Decr 0

*p<0.05 versus initial conditions

S-749 / © Mibelle Biochemistry S-749 / © Mibelle Biochemistry Mibelle AG Biochemistry, 5033 Buchs / Switzerland, Phone + 41 62 836 17 31, [email protected], www.mibellebiochemistry.com • Marketing Benefits anti-agingformulas • Liftingandfirming • Contouringmasks • Collagenboostingformulations • Tightening andcontouringserumsforthefaceneck Applications • Minimizesfinewrinklesanddeeperlines • Tightens facialcontours • Improves skindensity • Increases collagenandelastinexpression • Stimulatesexosomesignaling PhytoCellTec™ Goji for aV-shaped face Stem cellactivation PhytoCellTec™ Goji publication maybe reproduced ofMibelleAGBiochemistry. inany mannerwithoutthepriorwritten permission anddisclaims anyandallliabilityinconnection withyourproduct of thisinformation oritsuse.Nopartofthis erty rights.Mibelle AGBiochemistrywillnotassume anyexpressed orimplied liabilityinconnectionwithanyuse your obligationtocomplywithallapplicable lawsandregulations andtoobserveallthird partyintellectualprop statements are offered solelyforyourconsideration, investigationandverificationdonot relieve youfrom No legallybindingpromise orwarrantyregarding thesuitabilityofourproducts foranyspecificuseismade.Any containedinthis publication isprovidedThe information in good faithandisbased onourcurrent knowledge. Realized byscience naturally derivedcompoundsandprofound scientificknow-how. Inspired bynature – Mibelle Biochemistrydesignsanddevelopsinnovative, high-qualityactivesbasedon Innovating foryoursuccess • www.phytocelltec.ch • Sustainableproduction ofrawmaterial • Proven efficacy onmesenchymalstemcells • Applicationofexosomesignaling • Plantstemcellsfrom asuperfruit • Advancedstemcellcosmetics 3 Cosmetics –RawMaterialsActives rd PrizeattheBSBInnovationAward inthecategory -