J. Cosmet. Sci., 61, 311–324 (July/August 2010) 311 Evaluation of the effect of Thai breadfruit’s heartwood extract on the biological functions of fi broblasts from wrinkles JARUPA VIYOCH, SUPASIRI BURANAJAREE, FRANÇOIS GRANDMOTTET, SOPHIE ROBIN, DELPHINE BINDA, CÉLINE VIENNET, NETI WARANUCH, and PHILIPPE HUMBERT, Cosmetics and Natural Products Research Center (CosNat), Faculty of Pharmaceutical Sciences and Center of Excellence for Innovation in Chemistry, Naresuan University, Phitsanulok, 65000 Thailand (J.V., N.W.), Faculty of Pharmaceutical Sciences and Center for Innovation in Chemistry, Naresuan University, Phitsanulok, 65000 Thailand (S.B.), Engineering and Cutaneous Biology Team, University of Franche-Comté, Inserm UMR 645, IFR133, Besançon, 25000 France (F.G., S.R., D.B., C.V., P.H.), and Dermatology Department, St. Jacques Hospital, Besançon, 25030 France (P.H.). Accepted for publication April 23, 2010. Synopsis In previous studies, extract from Artocarpus incisus’s heartwood (breadfruit tree) had antioxidant and antimelanogenic activities. Here, we investigated the extract’s action on facial skin fi broblasts from wrin- kled skin and nonwrinkled skin biopsies, particularly in the production of type I procollagen and metallopro- teinase-1 (MMP-1) and in the reorganization of collagen fi bers. We found that the extract at a concentration of 50 μg/ml signifi cantly enhanced percent viability and proliferation of wrinkled-skin fi broblasts. Flow cytometry showed that a 3.6-fold increased proportion of the wrinkled-skin fi broblasts were in their cell cycle S-phase, indicating increased proliferation. Type I procollagen synthesis by wrinkled-skin fi broblasts was augmented by the extract. Nonwrinkled-skin fi broblasts had higher synthesis and were unaffected by the extract. MMP-1 secretion was greater for wrinkled-skin fi broblasts, but the extract decreased its secre- tion for both fi broblasts samples. Fibroblasts were incorporated in collagen lattice disks. Lattices with nonwrinkled-skin fi broblasts contracted uniformly by 56% after a three-day culture and the extract had little effect. However, wrinkled-skin fi broblast lattices failed to show appreciable contractions (to 12% after three days). But remarkably, the extract conferred an ability of the wrinkled-skin fi broblast lattices to fully contract (to 53%). This shows that wrinkled-skin fi broblasts have the ability to reorganize collagen but that the extract can reactivate this latent potential. Our fi ndings for the fi rst time reveal that A. in- cisus’s heartwood extract reversed the fi broblast defi ciencies in the metabolism and reorganization of colla- gen and may underlie a wrinkle treatment. Address all correspondence to Philippe Humbert.
JOURNAL OF COSMETIC SCIENCE 312 INTRODUCTION Aging is now generally regarded as a failure of an organism to repair tissue at the same rate as it is damaged (1,2). The changes occurring during aging are associated with al- terations in skin appearance due to loss of tensile strength, with wrinkle formation, in- creased fragility, and decreased epidermal moisture content. Among these effects, wrinkles are an important indicator of aging and an interesting fi eld in cosmetic dermatology. The appearance of wrinkles is related to an infl exibility of the skin coupled with a slackening of the dermis (3,4). This arises from a reduced production of collagen fi bers accompanied by the degeneration of the surrounding collagenous network (5–7). Resident fi broblasts are intimately involved in the decreased synthesis of collagens such as type I procollagen and an increase in the degradation of collagens. This is through increased collagenases such as matrixmetalloproteinase-1 (MMP-1), which occurs as fi broblasts age (8–10). In addition, aged fi broblasts lose their capacity to adhere and move over collagen fi bers, thereby limiting their ability to reorganize and reorient dermal tissue including collagen fi bers (11–13). For this reason, the improvement of fi broblast activities in terms of dermal tissue biosynthesis and reorganization may be useful for a variety of cosmetic and thera- peutic applications. It has been demonstrated that agents such as all-trans retinol (14–17) and vitamin C (18,19) can stimulate collagen production and suppress MMP activity. In addition, it has been reported that vitamin C and soy peptides can restore the capacity of fi broblasts to realign the collagen fi bers, which in turn improves locomotion, distribution, and adhe- sion of fi broblasts in the collagen matrix (11). These fi ndings indicate that aged-related fi broblast dysfunction is, at least in part, reversible. Thus it would be realistic to develop new agents whose activities are similar to those of retinol- and/or soy proteins that could be applied as cosmetics. In the cosmetics industry, there is demand for multifunctional and effi cacious products based on real innovation and aligned with trends in the cosmetics market. For example, natural products with antioxidant, antityrosinase and antiaging activities have been sought for the treatment of photoaging, unwanted skin pigmentation, and wrinkles. Recently, we found that diethyl either extracts using the heartwood from the Moraceae family, particularly Artocarpus incisus (breadfruit or Sa-kae in Thai) exhibited antioxi- dant activity in a dose-dependent manner using the DPPH assay (20). Furthermore, it has been reported that a crude extract as well as some purifi ed compounds isolated from A. incisus’s heartwood inhibited melanin production (20–22). In the present study, therefore we compared the effects of the extract on some biological functions of fi bro- blasts from nonwrinkled and wrinkled skin that were obtained from a biopsy of skin at the outer corner of the eye. The fi broblast functions assessed included the stimulatory effect on proliferation, production of type I procollagen, and the inhibitory effect on the production of the major collagen-degrading enzyme (MMP-1). In addition, we studied the effect of the extract on the ability of “wrinkled-skin fi broblasts” to reorga- nize collagen fi bers in a collagen lattice. The effect can be measured as a contraction of the collagen lattice, which thus indicates reorganization capacity (23,24). The results obtained from our study fi rst revealed the biological effects A. incisus’s heartwood ex- tract on human wrinkled-skin fi broblasts that were distinct from non-wrinkled skin cells. They also show that A. incisus’s heartwood extract could have potential cosmetic applications.
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