Development and mechanistic investigation of recombinant type III humanized collagen gel for mid-facial soft tissue repair

Abstract

Mid-facial depression is a key sign of facial aging, primarily caused by the loss of collagen leading to depletion of the extracellular matrix (ECM). However, the existing fillers for soft tissue augmentation have shown certain limitations in repairing mid-facial depression. Therefore, we herein report the development of a novel recombinant humanized type III collagen gel (C3Gel) through rational design and modification of a commercially available recombinant type III humanized collagen lyophilized fiber product. Both biological activity and tissue repair mechanisms of C3Gel were systematically evaluated in vitro and in vivo. C3Gel formed a dense fibrous structure around cells, significantly improving the ECM environment and providing strong support for cells, thereby promoting cell adhesion, migration, and proliferation. After injection of C3Gel into the dorsal region of rats, we observed a significant increase in the expression of type I collagen and elastin that improved tissue mechanical properties and elasticity. High-throughput RNA sequencing analysis revealed that C3Gel activated the integrin signaling pathway to improve binding between cells and ECM, resulting in the increased expression of downstream genes by activating the PI3K-Akt pathway which promoted the production of ECM components, such as collagen and laminin. At the same time, the expression of matrix metalloproteinases was inhibited to maintain ECM stability. Moreover, C3Gel is not carcinogenic in mice. Therefore, C3Gel demonstrates excellent biocompatibility and significant tissue repair ability, offering a safe, efficient, and long-term stable solution for mid-facial soft tissue augmentation, while providing new insights for other applications in regenerative medicine.

Graphical abstract

Keywords

Recombinant type III humanized collagen / Gel / Function / Mechanism of action / Stabilization / Biological Sciences / Biochemistry and Cell Biology

References

[1] LiQ, ShiB, ZhaoQ, ChenP, GuoZ, JinY, et al. . Expert consensus on facial rejuvenation in Chinese population. Chin J Med Aesthet Cosmet, 2020, 26011-7.

[2] LiuC, WangT. Progress in diagnosis and treatment of midface rejuvenation. Chin J Med Aesthet Cosmet, 2023, 290170-73.

[3] GuoJ, FangW, WangF. Injectable fillers: current status, physicochemical properties, function mechanism, and perspectives. RSC Adv, 2023, 133423841-23858.

[4] LafailleP, BenedettoA. Fillers: contraindications, side effects and precautions. J Cutan Aesthet Surg, 2010, 3116-19.

[5] KhaterR, AtanassovaPAgulloF. Autologous fat grafting—factors of influence on the therapeutic results. Current concepts in plastic surgery, 2012LondonIntechOpen183-210.

[6] TrinhLN, GuptaA. Non-hyaluronic acid fillers for midface augmentation: a systematic review. Fac Plast Surg, 2021, 3704536-542.

[7] TrévidicP, Kaufman-JanetteJ, WeinkleS, WuR, DhillonB, AntunesS, et al. . Injection guidelines for treating midface volume deficiency with hyaluronic acid fillers: the ATP approach (anatomy, techniques, products). Aesthet Surg J, 2022, 428920-934.

[8] Borzabadi-FarahaniA, MosahebiA, ZargaranD. A scoping review of hyaluronidase use in managing the complications of aesthetic interventions. Aesthet Plast Surg, 2024, 4861193-1209.

[9] Moers-CarpiMM, SherwoodS. Polycaprolactone for the correction of nasolabial folds: a 24-month, prospective, randomized, controlled clinical trial. Dermatol Surg, 2013, 393 Pt 1457-463.

[10] LiH, XuX, WuL, ChenX, AkhterH, WangY, et al. . Recent progress and clinical applications of advanced biomaterials in cosmetic surgery. Regen Biomater., 2023, 10: rbad005.

[11] QiaoS, PeiY, LanX. A new biomedicalmaterial of type A recombinant humanized collagen type III. Emerg Sci Technol, 2024, 1: 0009-0017.

[12] HuaC, ZhuY, XuW, YeS, ZhangR, LuL, et al. . Characterization by high-resolution crystal structure analysis of a triple-helix region of human collagen type III with potent cell adhesion activity. Biochem Biophys Res Commun, 2019, 50841018-1023.

[13] BoudkoSP, BachingerHP. The NC2 domain of type IX collagen determines the chain register of the triple helix. J Biol Chem, 2012, 2875344536-44545.

[14] YaoJ, YanagisawaS, AsakuraT. Design, expression and characterization of collagen-like proteins based on the cell adhesive and crosslinking sequences derived from native collagens. J Biochem, 2004, 1365643-649.

[15] MechlingDE, BachingerHP. The collagen-like peptide (GER)15GPCCG forms pH-dependent covalently linked triple helical trimers. J Biol Chem, 2000, 2751914532-14536.

[16] BoudkoS, FrankS, KammererRA, StetefeldJ, SchulthessT, LandwehrR, et al. . Nucleation and propagation of the collagen triple helix in single-chain and trimerized peptides: transition from third to first order kinetics. J Mol Biol, 2002, 3173459-470.

[17] HeinoJ. Cellular signaling by collagen-binding integrins. Adv Exp Med Biol, 2014, 819: 143-155.

[18] KechagiaJZ, IvaskaJ, Roca-CusachsP. Integrins as biomechanical sensors of the microenvironment. Nat Rev Mol Cell Biol, 2019, 208457-473.

[19] YounesiFS, MillerAE, BarkerTH, RossiFMV, HinzB. Fibroblast and myofibroblast activation in normal tissue repair and fibrosis. Nat Rev Mol Cell Biol, 2024, 258617-638.

Funding

Natural Science Foundation of Beijing Municipality(JQ24056)

RIGHTS & PERMISSIONS

The Author(s)