Triple Cross-Linked Hyaluronic Acid Based on Tri-Hyal Technique Has More Durable Effect on Dermal Renewal

Abstract

Background Hyaluronic acid (HA)-based fillers are applied to treat facial wrinkles and volume loss. Many efforts have been made to improve properties of HA to prolong the duration in aesthetic indications. A new cross-linking technique called “Tri-Hyal”, could make HAs to achieve desired rheological characteristics. HAs synthesized by Tri-Hyal are triple cross-linked and sustained-release, which could increase duration of promoting skin rejuvenation after injection. Purpose To evaluate the efficiency and persistence of HAs with Tri-Hyal on skin rejuvenation and further investigate underlying mechanisms, we compared the performances of cross-linked HA (AF) based on Tri-Hyal with another highly acceptable HA filler (Res) in vivo and in vitro. Methods Male BALB/c mice were divided into three groups, treated with AF, Res and vehicle, respectively. Skin biopsies were taken on day 0, 30, 90 and 180 after injection and hematoxylin and eosin (H&E), Masson’s trichrome (MT), immunohistochemical (IHC) stainings for CD31, TGF-β and MMP9 were performed. EdU incorporation, cell counting kit-8 (CCK-8), SA-β-Gal staining and activity were measured by biochemical analysis. RFP-GFP-LC3 adeno virus was used for autophagic flux detection. Protein levels of CD44, P62 and LC3I/II were detected by Western blot. Reactive oxygen species (ROS) level was detected by flow cytometry with DCFH-DA probe. Results The AF synthesized by Tri-Hyal showed persistent dermal structural correction without attenuation up to 6 months, which was illustrated by skin thickness, formation of elastic fibers and vascular density. Consistently, in fibroblasts the AF improved cell proliferation and slowed the senescent in vitro. Furthermore, it promoted cellular autophagy to reduce ROS level, which would account for its function in skin renewal. Conclusion The HA with Tri-Hyal could stimulate the production of extracellular matrix components more persistently than traditional HA fillers. In terms of mechanisms, it delayed senescence in dermal fibroblasts through reducing oxidative stress mediated by induction of autophagy.

A prospective multicenter clinical trial evaluating the efficacy and safety of a hyaluronic acid‐based filler with Tri‐Hyal technology in the treatment of lips and the perioral area

“…Several advantages have been discussed regarding this manufacturing technology. Exemplary, studies have suggested the minimization of cross‐linking agent that could reduce potential toxicity, the slow release of free HA which can promote extra‐cellular matrix production by fibroblasts and longer‐lasting stimulation of dermal growth 27–29 . However, more basic science research needs to be conducted to further define the molecular mechanisms by which soft‐tissue fillers manufactured using Tri‐Hyal technology improve skin quality.…”

Water Absorption and 3D Expansion of Different Injectable Hyaluronic Acids

“…The ART FILLER® line is a group of fillers produced with Trihyal technology, meaning they have long chain, very-long chain, and free non-crosslinked HA in their composition [34,35]. The Universal filler was chosen as the control group, as it is the most used one out of the five and the most versatile.…”