In Vitro Evaluation of the Sensitivity of a Hyaluronic Acid PEG Cross-Linked to Bovine Testes Hyaluronidase

Nicola Zerbinati; Torello Lotti; Damiano Monticelli; Virginia Martina; Giovanna Cipolla; Edoardo D'Este; Alberto Calligaro; Roberto Mocchi; Cristina Maccario; Sabrina Sommatis; Jacopo Lotti; Uwe Wollina; Georgi Tchernev; Katlein FranÃ§a

doi:10.3889/oamjms.2018.046

Authors

Nicola Zerbinati Universita degli Studi dell'Insubria Dipartimento di Scienze Chirurgiche e Morfologiche, Varese
Torello Lotti University G. Marconi of Rome, Dermatology and Venereology, Rome
Damiano Monticelli Universita degli Studi dell'Insubria Dipartimento di Scienza e Alta Tecnologia, Como
Virginia Martina Matex Lab, Brindisi
Giovanna Cipolla UniversitÃ di Pavia - Centro Medico Polispecialistico, Pavia
Edoardo D'Este UniversitÃ di Pavia - Centro Medico Polispecialistico, Pavia
Alberto Calligaro University of Pavia - Department of Public Health, Experimental and Forensic Medicine, Pavia
Roberto Mocchi UB - CARE S.r.l. - Spin - off University of Pavia, Pavia
Cristina Maccario UB - CARE S.r.l. - Spin - off University of Pavia, Pavia
Sabrina Sommatis UB - CARE S.r.l. - Spin - off University of Pavia, Pavia
Jacopo Lotti University of Rome â€œG. Marconiâ€ - Department of Nuclear, Subnuclear and Radiation Physics, Rome
Uwe Wollina StÃ¤dtisches Klinikum Dresden, Department of Dermatology and Venereology, Dresden, Sachsen
Georgi Tchernev Medical Institute of Ministry of Interior (MVR), Department of Dermatology, Venereology and Dermatologic Surgery, Sofia
Katlein FranÃ§a University of Miami School of Medicine, Department of Psychiatry & Behavioral Sciences, Miami, Florida

DOI:

https://doi.org/10.3889/oamjms.2018.046

Keywords:

Hyaluronic acid, Sensitivity to hyaluronidase, N-acetyl-D-glucosamine, Neauvia Intense, Poly ethylene glycol

Abstract

Neauvia Intense is biocompatible, injectable hyaluronic acid (HA) filler PEG cross-linked for facial soft-tissue augmentation that provides volume to tissues. The aim of the present study is to evaluate the sensitivity of Neauvia Intense in hyaluronidase from bovine testes in a time-course analysis. The test is based on the colourimetric determination of the N-acetyl â€“ D - glucosamine (NAG) released by the hyaluronidase in standardised conditions. The in vitro conditions involve the treatment of Neauvia Intense with a known concentration of the enzyme (6080U/ml). The NAG content was determined at different times to assess the kinetics of the degradation (1h, 3h, 6h, 24h, 48h, 72h, 120h, and 168h); the Ehrlichâ€™s reagent was used for the colourimetric quantification, by the method described by Reissing and colleagues. The intensity of the violet colour developed after the chemical reaction was proportional to the NAG present in each sample. A microplate reader at 585 nm read the absorbance. The amount of NAG released by the product was proportional to the time of incubation with bovine hyaluronidase, reaching a plateau after 168 hours.

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