74 MARINE INGREDIENTS
Material and methods The complex studied named Okinacea®
a (Sea
grape) contains water (65%), Caulerpa lentillifera extract (31%) and hydrolyzed rice bran protein (4%). It is safe for cosmetic applications following the results of Patch - Het Cam – Ames and sensitizing potential testing. INCI name is: Water (and) Caulerpa
Lentillifera Extract (and) Hydrolyzed Rice Bran Protein.
The thallus of Caulerpa lentillifera consists of horizontal irregularly branched stolons bearing numerous erect fronds which are densely covered by subspherical short ramelli. Each ramulus consists of a short stalk and a globose tip, 1-3 mm in diameter (Fig 1). The distinct constriction between the tip of the stalk and the base of the globose tip is a characteristic of this species (Fig 2).
Caulerpa lentillifera used for the manufacturing of this active came from controlled cultures in Asia.
In vitro study The synthesis of procollagen 1 has been quantified on the supernatant of the cultures of normal human fibroblasts after 48 H cultivation in the presence of 2% active by using the kit ‘Procollagen type 1C- peptide (PIP) EIA Kit - ref. MK101 – TANAKA’. The standard was vitamin C 50 µg/ml. – positive control. The total protein content was evaluated according to the bicinchoninic acid (BCA) method.
Ex vivo study and immunolabelling All ex vivo studies have been performed on 24 skin explants obtained from an abdominal plastic surgery of a healthy Caucasian woman (53 years old). They are cultured in the specific survival explants medium BEM. 2 mg of a formulation (Carbopol gel) containing 2% of the active mixture were applied on the skin stripes at the following times: Day 0, Day 2, Day 5 and Day 7. The results are compared with untreated explants. Explants were taken off at D0 and D9. Each explant was cut into
a b
Figure 3: Immunolabelling of collagen IV in human skin explants at Day 9. A Control explants without any treatment. The labelling appears as a scalloped band relatively clear and relatively regular along the DEJ. B Treated explants with 2% sea grape. The marking is distinct and regular along the DEJ. It also appears in the basal keratinocytes
a b
Figure 4: Immunolabelling of laminin V in human skin explants at Day 9. A Control explants without any treatment. The labelling appears as a scalloped band relatively clear and relatively regular along the DEJ. B Treated explants with 2% sea grape. The marking is distinct and regular along the DEJ. It also appears in the basal keratinocytes.
two parts: one half was fixed in ordinary Bouin solution for previous morphological analysis while the other half was frozen at - 75°C for immunostaining.
Specific immunolabellings of collagen IV, laminin-5, GAGs and collagen I were performed on frozen cryostat cut tissues thanks to specific monoclonal antibodies and revealed by FITC (fluorescein isothiocyanate). Cells nuclei are then stained with propodium iodide. Optical observations have been performed by
b
using a microscope Leica type DMLB equipped with a camera Olympus DP 72.
In vivo study The in vivo study has been performed on 25 healthy women (age from 40 to 59). A formulation with Okinacea (5%) as a unique active ingredient has been applied twice a day for 28 days on crow’s feet area. The measurement of the skin roughness and 3D pictures have been performed by fringes projection using a GFM PRIMOS®
device.
Results and discussion Results presented here after show that sea grape was able to prevent some anti- ageing damage.
Figure 5: Immunolabelling of GAGs in human skin explants at Day 9. A Control explants without any treatment. GAGs present along the DEJ form a pink-violet band, regular and thin in the papillary dermis. B Treated explants with 2% sea grape. The pink-violet staining is regular and variously important.
PERSONAL CARE EUROPE
Sea grape strengthens the dermal epidermal junction (DEJ) The DEJ serves various functions; e.g. epidermal-dermal adherence, mechanical support for the epidermis. It also controls proliferation and differentiation of basal cells in the epidermis. It maintains the polarity of the epidermis ensuring the cohesion and exchanges between the two major skin compartments. The ageing of the skin comes with significant physiological changes to the skin, especially at the DEJ
June 2019
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