Mondon Philippe, Hillion Mélanie, Peschard Olivier, Andre Nada, Marchand Thibault, Doridot Emmanuel, Feuilloley Marc Gj, Pionneau Cédric, Chardonnet Solenne
SEDERMA, Le Perray-en-Yvelines, France.
LMSM EA4312, University of Rouen, Evreux, France.
J Cosmet Dermatol. 2015 Jun;14(2):152-60. doi: 10.1111/jocd.12135. Epub 2015 Mar 27.
This study was conducted to establish a new methodology for evaluating elements of dermal extracellular matrix (ECM), of epidermal-dermal junction (EDJ), and effects of molecules which can modulate their synthesis. This methodology is based on matrix-assisted laser desorption/ionization mass spectrometric imaging (MALDI-MSI). In vivo reflectance confocal microscopy (in vivo RCM) and echography were also used. Using immunohistochemistry methods on explants, age-related modification data were obtained for selected dermal ECM and EDJ proteins (collagen I, collagen IV, collagen VII, collagen XVII, nidogen I, decorin/decorunt) and used as reference for MALDI-MSI studies. A methodology was developed with MALDI-MSI to map epidermis and dermis proteins. Then MALDI-MSI was used to study age modifications. In vivo RCM and high-frequency ultrasounds were used to evaluate ECM and EDJ undulation modifications caused by aging. Anti-aging molecule evaluations were performed with a blend of palmitoyl oligopeptide and palmitoyl tetrapeptide-7. Immunohistochemistry studies demonstrated that the selected proteins were found to be less abundant in aged group explants vs. young group except for decorin. MALDI-MSI studies correlated the results obtained for decorin. In vivo RCM measurements indicated a decrease of EDJ undulation depth with age and ECM modifications in the upper part of dermis. Echography demonstrated that the peptide blend reduced subepidermal low-echogenic band thickness and improved its density. In vivo RCM studies indicated that the peptides improved the ECM structure vs. placebo. This preliminary MALDI-MSI study raised some technical difficulties that were overcome. Further studies will be conducted to identify more proteins and to demonstrate the interest of this method for cosmetic evaluations.
本研究旨在建立一种新的方法，用于评估真皮细胞外基质（ECM）、表皮-真皮连接（EDJ）的成分，以及能够调节其合成的分子的作用。该方法基于基质辅助激光解吸/电离质谱成像（MALDI-MSI）。还使用了体内反射共聚焦显微镜（体内RCM）和超声检查。通过对植块进行免疫组织化学方法，获得了所选真皮ECM和EDJ蛋白（I型胶原、IV型胶原、VII型胶原、XVII型胶原、巢蛋白I、核心蛋白聚糖/双糖链蛋白聚糖）与年龄相关的修饰数据，并将其用作MALDI-MSI研究的参考。开发了一种利用MALDI-MSI绘制表皮和真皮蛋白图谱的方法。然后使用MALDI-MSI研究年龄相关的修饰。使用体内RCM和高频超声评估衰老引起的ECM和EDJ波动变化。使用棕榈酰寡肽和棕榈酰四肽-7的混合物进行抗衰分子评估。免疫组织化学研究表明，除核心蛋白聚糖外，老年组植块中所选蛋白的含量低于年轻组。MALDI-MSI研究使核心蛋白聚糖的结果具有相关性。体内RCM测量表明，随着年龄增长，EDJ波动深度降低，真皮上部的ECM发生改变。超声检查表明，该肽混合物降低了表皮下低回声带的厚度并改善了其密度。体内RCM研究表明，与安慰剂相比，这些肽改善了ECM结构。这项初步的MALDI-MSI研究提出了一些克服的技术难题。将进行进一步研究以鉴定更多蛋白质，并证明该方法在化妆品评估中的价值。
