Advanced quantitative proteomics to evaluate molecular effects of low-molecular-weight hyaluronic acid in human dermal fibroblasts

• Published in: Journal of pharmaceutical and biomedical analysis 2020-06, Vol.185, p.113199, Article 113199
• Main Authors: Radrezza, Silvia, Baron, Giovanna, Nukala, Sarath Babu, Depta, Gabriele, Aldini, Giancarlo, Carini, Marina, D’Amato, Alfonsina
• Format: Article
• Language: English
• Subjects: Cell Line; Collagen - biosynthesis; Consumer Product Safety; Cosmetics - adverse effects; Cosmetics - chemistry; Cosmetics - standards; Dermal fibroblasts; Extracellular Matrix - drug effects; Feasibility Studies; Fibroblasts - drug effects; Fibroblasts - metabolism; Humans; Hyaluronic Acid - adverse effects; Hyaluronic Acid - chemistry; Hyaluronic Acid - standards; Low-molecular-weight hyaluronic acid; Mass spectrometry; Mass Spectrometry - methods; Molecular Weight; Networking; Proteoglycans - biosynthesis; Proteomics - methods; Quantitative proteomics; Skin - cytology
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2020.113199

[Display omitted] •Low-molecular-weight hyaluronic acid influenced intra- and extra-cellular proteome.•Label free proteomics allowed a complete vision of molecular impact on fibroblast.•Safety and benefits for dermal use were confirmed. Hyaluronic acid (HA) is physiologically synthesized by several human cells types but it is also a widespread ingredient of commercial products, from pharmaceuticals to cosmetics. Despite its extended use, the precise intra- and extra-cellular effects of HA at low-molecular-weight (LWM-HA) are currently unclear. At this regard, the aim of this study is to in-depth identify and quantify proteome’s changes in normal human dermal fibroblasts after 24 h treatment with 0.125, 0.25 and 0.50 % LMW-HA (20−50 kDa) respectively, vs controls. To do this, a label-free quantitative proteomic approach based on high-resolution mass spectrometry was used. Overall, 2328 proteins were identified of which 39 significantly altered by 0.125 %, 149 by 0.25 % and 496 by 0.50 % LMW-HA. Protein networking studies indicated that the biological effects involve the enhancement of intracellular activity at all concentrations, as well as the extracellular matrix reorganization, proteoglycans and collagen biosynthesis. Moreover, the cell’s wellness was confirmed, although mild inflammatory and immune responses were induced at the highest concentration. The more complete comprehension of intra- and extra-cellular effects of LMW-HA here provided by an advanced analytical approach and protein networking will be useful to further exploit its features and improve current formulations.