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. 2018 Jul 11:11:339-346.
doi: 10.2147/CCID.S156740. eCollection 2018.

A novel animal model for residence time evaluation of injectable hyaluronic acid-based fillers using high-frequency ultrasound-based approach

Filomena Merola  1 , Mario Scrima  1 , Carmela Melito  1 , Antonio Iorio  1 , Claudio Pisano  2 , Andrea Maria Giori  3 , Angela Ferravante  1
Affiliations

A novel animal model for residence time evaluation of injectable hyaluronic acid-based fillers using high-frequency ultrasound-based approach

Filomena Merola et al. Clin Cosmet Investig Dermatol. .

Abstract

Background: Hyaluronic acid (HA)-based devices are among the most popular filler agents for skin rejuvenation. One of the principal goals is the improvement in residence time of HA-based products, to increase their performance and reduce frequency of the treatment. So, understanding fillers, behavior after subcutaneous injection is a fundamental aspect for discovery and optimization of new products. Current in vivo approaches to detect/quantify injected HA fillers are not always well optimized or easy to apply.

Objective: To develop more efficacious and noninvasive diagnostic tools to make a quantitative evaluation of the degradation of fillers in a small animal model.

Materials and methods: We evaluated the residence time of different HA-based fillers, fluorescein-labeled and not, injected subcutaneously in mice. Volumes of fillers were monitored through high-frequency ultrasound (HF-US) method while fluorescence intensity through the well-established fluorescence living imaging method. To confirm the effectiveness of HF-US, obtained volumetric measurements were compared with fluorescence intensity values.

Results: Both the presented methods revealed the same degradation kinetics for the tested products.

Conclusion: The two used methods are fully comparable and quantitatively accurate. The presented approach has been proved to be noninvasive, sensitive, and reproducible.

Keywords: HF-US; animal model; dermal filler; hyaluronan; hyaluronic acid; ultrasound.

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Conflict of interest statement

Disclosure Andrea Maria Giori is an employee of IBSA Farmaceutici Italia, a pharmaceutical company based in Lodi, Italy. Filomena Merola, Mario Scrima, Carmela Melito, Antonio Iorio, and Angela Ferravante are employees of Bouty SpA., a pharmaceutical company based in Milan, Italy. Claudio Pisano is a consultant of Bouty SpA. The authors report no other conflicts of interest in this work.

Figures

Figure 1
Figure 1
(A) Subcutaneous injection of test items. On the dorsal region of the mouse are visible the two mounds generated by injection of test items. (B) Bi-dimensional image. Sonographic image of the subcutaneous injection of dermal filler in the dorsal area. The filler appears as anechogenic area. (C) Three-dimensional image of a dermal filler. The figure shows a 2D ultrasound image and the 3D reconstruction of the volume occupied by the filler.
Figure 2
Figure 2
(A) Volumetric degradation of fillers monitored through HF-US. (B) Representative images of degradation kinetics of CL-HA by HF-US. Abbreviations: HF-US, high-frequency ultrasound; CL-HA, cross-linked hyaluronic acid; L-HA, linear hyaluronic acid; L-HA-F, fluorescein-labeled linear hyaluronic acid.
Figure 3
Figure 3
Volumetric degradation of hyaluronic acid products. (A) Degradation kinetics of L-HA-F 0.8% monitored through HF-US. (B) Degradation kinetics of (L-HA + L-HA-F) 0.7% monitored through HF-US. (C) Degradation kinetics of L-HA-F 0.8% (left flank) and (L-HA + L-HA-F) 0.7% (right flank) monitored through FLI. Abbreviations: HF-US, high-frequency ultrasound; L-HA, linear hyaluronic acid; L-HA-F, fluorescein-labeled linear hyaluronic acid; FLI, fluorescence living imaging.
Figure 4
Figure 4
Graphical representation of the statistical correlation (P<0.01) existing between volumes of fillers and absorbance values obtained through HF-US and FLI techniques for L-HA-F 0.8% (A) and (L-HA + L-HA-F) 0.7% (B). Abbreviations: HF-US, high-frequency ultrasound; L-HA, linear hyaluronic acid; L-HA-F, fluorescein-labeled linear hyaluronic acid; FLI, fluorescence living imaging.
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