Acj acs 80 4

Niacinamide (NIA), k-C RG, polyvinylpyrrolidone (PVP), jojoba oil, tween 80, oleic acid, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) powder, k-carrageenan, chloroform, jojoba oil, trypsin, and phosphate-buffered saline (PBS) were supplied by Sigma-Aldrich (Alabaster, AL, USA). l-α-phosphatidylcholine (EPC) was purchased from Avanti Polar Lipids (Alabaster, AL, USA). Ethanol absolute (≥99.8%) and methanol (≥99.8% HPLC grade) were obtained from Fisher Chemical (Thermo Fisher Scientific, Loughborough, UK). Double-deionized water was provided by an ultra-pure water system (Arium Pro, Sartorius AG, Gottingen, Germany). The porcine ears were acquired in a local slaughterhouse (Porto, Portugal). Reagents were weighted in a digital analytical balance Kern ACJ/ACS 80-4 (Kern & Sohn; Balingen, Germany). The pH measurements were achieved using a Crison pH meter GLP 22 with a Crison 52-02 tip (Crison; Barcelona, Spain).

Sodium alginate was purchased from ACROS Organics^TM (Thermo Fisher Scientific, Massachusetts, MA, USA). Calcein, k-CRG, RV tartrate, and Dulbecco’s phosphate-buffered saline (PBS) (10×) were acquired from Sigma-Aldrich (St. Louis, MO, USA). Exelon^® patches were acquired from Novartis. Acetonitrile was acquired from Fisher Scientific (Hampton, NY, USA). Silicone moulds were obtained from Micropoint Technologies (Pioneer junction, Singapore). Fresh porcine ears obtained from different adult animals were purchased in a local slaughter (Porto, Portugal). Double-deionised water was provided by an ultra-pure water system (Arium Pro, Sartorius AG, Gottingen, Germany). The reagents were weighted in a digital analytical balance Kern ACJ/ACS 80-4 (Kern & Sohn; Balingen, Germany). pH measurements were achieved using a JENWAY 550 pH meter (Staffordshire, UK). Texture analysis was performed using a TA.XT2 Texture Analyser (Stable Micro Systems Ltd., Haslemere, UK). SEM and confocal fluorescence microscopy studies were performed at the Laboratory for Scanning Electron Microscopy and X-Ray Microanalysis and High-Resolution Microscopy Unit, respectively, both at CEMUP (Materials Centre of the University of Porto, “Centro de Estudos de Materiais da Universidade do Porto”). Histological samples have been processed in the histological service of “ICBAS, Universidade do Porto”.

Fucoidan extracted from Fucus vesiculosus (92.9% fucoidan phytonutrient content, high molecular weight, code 5619002900, batch FVF2016507) was a kind gift from Kraeber & Co. GMBH (Ellerbek, Germany). Sodium alginate (from Lessonia nigrescens, code 177775000, lot A0376873, sulfated ash 34.16% (on dried substance)) was purchased from ACROS OrganicsTM (Thermo Fisher Scientific, Waltham, MA, USA). (−)-Menthol was acquired from Fluka (Buchs, Switzerland). Bergamot oil was purchased from Pranarôm (Ghislenghien, Belgium). L-Linalool, (−)-β-pinene, calcein, 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES) hemisodium salt, and trypsin (from the porcine pancreas) were acquired from Sigma-Aldrich (St. Louis, MO, USA). The porcine ears were purchased at a local slaughter (Porto, Portugal). Double-deionized water was provided by an ultra-pure water system (Arium Pro, Sartorius AG, Gottingen, Germany). The reagents were weighted in a digital analytical balance Kern ACJ/ACS 80-4 (Kern & Sohn; Balingen, Germany). All other reagents were analytical grade and were used without any further purification.