Hyaluronic acid-cross-linked filler stimulates collagen type 1 and elastic fiber synthesis in skin through the TGF-β/Smad signaling pathway in a nude mouse model

J Plast Reconstr Aesthet Surg. 2019 Aug;72(8):1355-1362. doi: 10.1016/j.bjps.2019.03.032. Epub 2019 Apr 11.


Compared to pure hyaluronic acid filler, cross-linked hyaluronic acid (HAc) exhibits superior biocompatibility and longevity as a dermal filler. We previously developed composite HAc-hydroxyapatite (HAp) fillers. Herein, we systematically compared the protein-level increase and gene expression between HAc-micro-HAp and HAc-nano-HAp in mice and determined the mechanisms underlying the biological responses to HAc and HAp. Five-week-old female BALB/c-nude mice were classified into five groups: normal skin, Radiesse, Restylane, HAc-nano-HAp, and HAc-micro-HAp. Fillers (200 μl) were injected to evenly fill the back of mice. Skin biopsies were performed to investigate collagen and elastic fiber synthesis after filler injections. Western blot analysis, real-time polymerase chain reaction analysis, and immunohistochemistry were performed to investigate protein and gene expression changes. Organ (liver, lung, spleen, and kidney) toxicity of HAc-nano-HAp was determined by hematoxylin and eosin staining after 12 weeks. Protein and gene expression analyses indicated that, compared with pure fillers, HAc-nano-HAp and HAc-micro-HAp hydrogels preferentially promoted collagen and elastic fiber formation through the TGF-β pathway. The composite fillers also exhibited no evidence of organ toxicity. HAc-HAp filler might play an important role in collagen and elastic fiber regeneration. HAc filler stimulates collagen type 1 and elastic fiber synthesis through the TGF-β/Smad pathway. The role of HAc-HAp composite fillers in photoaging in animal models and their effects on skin, including elasticity and tensile strength, should be investigated.

Keywords: Collagen type 1; Dermal filler; Elastic fiber; HAc-micro-HAp; HAc-nano-HAp; TGF-β/Smad pathway.

MeSH terms

  • Animals
  • Biocompatible Materials / pharmacology
  • Collagen Type I / biosynthesis*
  • Collagen Type I / genetics
  • Dermal Fillers / pharmacology*
  • Disease Models, Animal
  • Elastic Tissue / metabolism*
  • Female
  • Gene Expression
  • Humans
  • Hyaluronic Acid / pharmacology*
  • Hydrogels
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Skin / drug effects*
  • Skin / metabolism
  • Smad Proteins / metabolism*
  • Transforming Growth Factor beta / metabolism*


  • Biocompatible Materials
  • Collagen Type I
  • Dermal Fillers
  • Hydrogels
  • Smad Proteins
  • Transforming Growth Factor beta
  • Hyaluronic Acid