![]() Depending on the integrity of the HA-filler the fluorescent dye remained either inclosed in the gel, resulting in high fluorescence intensity, or the dye was released and diluted in the surrounding medium, resulting in a decrease in fluorescence intensity. Next, the now fluorescent gel was placed in a larger volume of 350 µl of serum-free keratinocyte medium (SFM, Thermo Fisher Scientific) and wells were placed in a time-lapse videomicroscopy workstation (Zeiss Axiovert 200 M, Carl Zeiss Microscopy GmbH, Göttingen, Germany 37 ☌, 5% CO 2). First, 50 µl of each filler were mixed with 10 µl of a green fluorescent cell linker dye (PKH67, Sigma) in addition to 10 units (U/ml) of bovine testicular hyaluronidase (HYAL Hylase “Dessau”, Riemser Pharma, Greifswald, Germany), which represents the standard hyaluronidase used in Germany, or an equal volume of NaCl and placed in 24-well plates (BD Bioscience). Taking in account the potential risks associated with filler injections, the proven degradability of an HA-filler by hyaluronidase can be regarded as “safety-feature” and potential competitive edge over other manufacturers of dermal fillers.Īgainst this background, we here set out to systematically analyze the degradability of three commonly used HA-fillers by bovine hyaluronidase in a time dependent manner, using a novel standardized video-microscopic in vitro approach.įor our analysis we used three comparable commercially available HA-fillers, Belotero Balance Lidocain (BEL monophasic double cross-linked, BDDE, HA 22.5 mg/ml, with lidocaine Merz Pharmaceuticals GmbH, Frankfurt, Germany manufactured by ANTEIS SA, Geneva, Switzerland), Emervel classic (EMV cross-linked, biphasic, sizing, BDDE, HA 20.0 mg/ml, lidocaine Galderma manufactured by Q-Med AB, Uppsala, Sweden), and Juvederm Ultra 3 (JUV cross-linked “Hyalcross”, BDDE, HA 24.0 mg/ml, lidocaine Pharm-Allergan, Irvine, CA, USA). A difference in or even resistance to “degradability” may be attributed to the concentration of HA in the filler, the degree of cross-linking, and/or its cohesive properties. Despite the availability of hyaluronidase, it is controversially discussed whether all HA-fillers can be degraded by hyaluronidase assimilably effective. Since the timely infiltration of hyaluronidase may degrade HA-fillers and may rescue from more severe vascular complications, the immediate availability of hyaluronidase is regarded a necessity for every physician that injects HA. The availability of a specific antidote, hyaluronidase, for the management of complications of filler treatments is one major reason for the preferred use of HA-based fillers over other injectable fillers, such as calcium hydroxylapatite (CHA) or poly- l-lactic acid. Potential complications of filler treatments range from unaesthetic overcorrections, Tyndall effect or lower eyelid edema following tear-trough augmentation, to granulomas, infections, up to tissue necrosis or even blindness due to vascular occlusions. Today, the injection of reversible HA-based dermal fillers is regarded as gold standard for tissue augmentation, deep skin hydration or facial recontouring. A decrease in the skin’s HA content is considered as main characteristic of skin aging. Hyaluronic acid (hyaluronan, HA) is a non-sulfated glycosaminoglycan (GAG) and an essential part of the skin's extracellular matrix (ECM). Time-lapse microscopy enables systematically, standardized, comparative in vitro analyses of the interactions of hyaluronidase and HA-fillers. No effect of hyaluronidase was observed for JUV. Degradation was measurable at 5 h (BEL) and 7 h (EMV), respectively significance was reached at 14 h (BEL) and 13 h (EMV). Hyaluronidase showed a significant degradation of the HA-fillers BEL and EMV. HA-filler plus control, quantified by computer-assisted image analysis (ImageJ). The degradation of HA-fillers was assessed as decrease in fluorescence intensity of HA-filler plus hyaluronidase vs. MethodsĬomparable HA-fillers, Belotero Balance Lidocaine (BEL Merz), Emervel classic (EMV Galderma) and Juvederm Ultra 3 (JUV Allergan), were incubated with a fluorescent dye and bovine hyaluronidase (HYAL Hylase “Dessau”, Riemser) or control (NaCl) and monitored by time-lapse videomicroscopy. To analyze the interactions of HA-fillers and hyaluronidase in a time-dependent manner using a novel standardized in vitro approach. Yet, up to date there are only few studies that have systematically assessed the degradability of different HA-fillers by hyaluronidase. The “off-label” use of hyaluronidase is regarded as gold standard for the management of HA-filler-associated complications. Hyaluronidase is a hyaluronic acid (HA) metabolizing enzyme, which is approved as an adjuvant for infiltration anesthesia.
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