Introduction Hyaluronic acid (HA) is a high‐molecular‐weight polysaccharide composed of glucuronic acid and N‐acetylglucosamine. HA constitutes a major component of the extracellular matrix (ECM) of the skin. With aging, the amount of HA in skin decreases, which implies a loss of skin moisture causing wrinkles, sagging and aging of the skin. Oral supplementation with HA has been widely studied in order to provide an anti‐aging effect on the skin. There are mainly two types of HA: HA of fermentation and HA of extraction from a natural source. Dermial® is a HA matrix ingredient composed of HA and other naturally occurring components such as, dermatan sulphate and collagen, which have been shown to exert beneficial effects for skin health. This study was performed to determine the comparative in‐vitro effects of Dermial®, pure HA from extraction (HA‐E), and pure HA from fermentation (HA‐F). Methods The effect of the HA products on cell proliferation was assessed in Human Dermal Fibroblasts (HDF) and in Human Epidermal Keratinocytes (HEK) using the method bromodeoxyuridine (BrdU) incorporation, detected by ELISA. The cell migration was evaluated in HDF and in HEK using the Oris™ Cell Migration Assay. The inductive capacity on ECM protein synthesis was evaluated by immunolocalization‐ELISA. Results The results showed that the rate of cell proliferation in HDF with respect to untreated HDF was significantly higher in Dermial® showing a dose‐dependent response and reaching a proliferation rate of 1.81 (± 27) times with the highest concentration tested (1mg/mL) in comparison to HA‐E and HA‐F. Regarding HEK cell proliferation, Dermial® showed a significant capacity to induce this mechanism, increasing a rate of 85‐95% at 25 and 50 µg/mL. HA‐E and HA‐F showed similar effects, with significant increases on HEK cell proliferation (88% increase at 50 µg/mL of HA‐E and 100% at 25 µg/mL of HA‐F). Dermial®, HA‐E and HA‐F induced the migration of HDF with significant increases equal to or greater than 50%. However, only Dermial® showed a significant stimulatory effect on HEK migration, showing an increase of 102.77% (± 40.86) at 200 µg/mL. Dermial® showed a significant effect on the production of type I collagen, with an increase of 48.04% (± 11.49%) at 1 mg/mL. The quantification of type III collagen showed a significant effect of Dermial® and HA‐F, with increases of 77% for 1mg/mL Dermial® and 89‐118% for 50 µg/mL and 1 mg/mL HA‐F. Regarding the quantification of elastin, all evaluated products showed a significant effect, with increases of 17‐38%. Only Dermial® showed significant effect on the production of three proteins tested. Conclusions The overall results showed a superior effect of HA matrix ingredient (Dermial®) as compared to pure HA, either HA‐E or HA‐F, on HDF cell proliferation, on HEK cell migration and in the stimulation of the synthesis of type I collagen. Dermial® presents better regenerative properties of the dermis and epidermis, in addition to stimulating the synthesis of ECM proteins, favouring the maintena...
Introduction Dermatan sulfate (DS) is a linear polysaccharide classified as a sulfated glycosaminoglycan (GAGs). GAGs form an integral part of the skin and make up 0.1–0.3% of total skin weight. DS is covalently attached to the core proteins to form proteoglycans (PGs). PGs predominates in dermis presenting higher level of DS than other sulfated GAGs. The intrinsic aging processes of the skin entails the reduction of total sulfated GAGs levels causing a thinning of the dermis and modifying the quality of the skin. DS possess strong biological activity that can be modulated to improve skin quality. This study was performed to determine the effects of DS on Human Dermal Fibroblasts (HDF) to evaluate its anti‐aging and regenerative properties. Methods Human Dermal Fibroblasts (HDF) were isolated from a sample of the foreskin of a neonatal caucasian donor (Lonza). The effect of the DS on cell proliferation was assessed at 24 h and 48 h in HDF, using the method bromodeoxyuridine (BrdU) incorporation, detected by ELISA. The cell migration was evaluated in HDF at 48 h using the Oris™ Cell Migration Assay (Platypus Technologies). To monitor the migration progress, the cells were labelled with calcein. The wound healing capacity of DS was evaluated in HDF using the scratch test. The inductive capacity on Extracellular matrix proteins (ECMp) synthesis was evaluated by immunolocalization‐ELISA quantifying type I and type III collagen and elastin. Results The results showed that DS induced HDF proliferation after 24 h and 48 h of treatment at all tested concentrations. A dose‐dependent increase in proliferation was observed after 48 h of treatment of these cells, reaching a maximum increase of 5.10 (± 0.82) fold at the highest concentration tested (5 mg/mL) compared to uHDF. Regarding HDF cell migration, no significant differences were observed at 24 h after treatment with DS. The results showed a significant improvement in the in vitro wound healing capacity of HDF treated with 0.25 and 1 mg/mL of DS, with an increase in the healed area of 30.28% (± 4.27%) and 23.76% (± 8.87%) respectively, compared to uHDF. Regarding the ECMp synthesis‐stimulating capacity of DS, the results showed a significant increase of both type III Collagen (50.98 ± 34.48% increase) and Elastin (32.25 ± 2.61%) levels in HDF treated for 72 h with 1 mg/mL of DS compared to uHDF. HDF treated with 0.25 mg/mL also showed higher type I Collagen production than uHDF, although the increase (30.96 ±7.04%) was not statistically significant. Conclusions The results showed that DS improves the proliferation and regeneration of HDF present in dermis. In addition, DS increases the synthesis of ECMp as collagen type III and elastin, promoting the maintenance and functionality of the dermis. All these properties support the regenerative and anti‐aging activity of DS acting against intrinsic aging in dermis.
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