In Vitro Evaluation of the Biosafety of Hyaluronic Acid PEG Cross-Linked with Micromolecules of Calcium Hydroxyapatite in Low Concentration
Keywords:hyaluronic acid, biosafety, calcium hydroxyapatite, human keratinocytes, Neauvia Stimulate
OBJECTIVE: Neauvia Stimulate is biocompatible, injectable hyaluronic acid (HA) filler (26 mg/ml) PEG cross-linked with 1% of calcium hydroxyapatite (CaHA) for facial soft-tissue augmentation that provides volume to tissues, followed by process of neocollagenesis for improving skin quality.
AIM: The aim of the present study is to evaluate the biosafety of the product (Lot. 160517-26-1/2 PEG) on human keratinocytes cultured in vitro.
MATERIAL AND METHODS: The experimental model proposed, despite being an in vitro system, allows the derivation of useful information to predict the possible activity of the product in further in vivo application. Human keratinocytes (HaCaT cells) were treated with the product for 24h at increasing concentrations of product respect to control (untreated cells).
RESULTS: The biosafety of the product to be tested has been evaluated performing different methods: MTT test, NRU test, Kenacid Blue assay. Moreover, any possible effect on the structure, morphology, and viability of cells has been evaluated.CONCLUSION: In conclusion, the results obtained by the different methods show that the product Neauvia StimulateÂ® does not cause any cytotoxic effect and does not affect the correct structure and morphology of cells cultures.
Larsen NE, Leshchiner E, Pollak CT, Balasz EA, Piacquadio E. Evaluation of Hylan B (hylan gel) as soft tissue dermal implants. Proc Mat Res Soc (Spring Meeting, San Francisco, CA). 1995; 394:193-7. https://doi.org/10.1557/PROC-394-193
Larsen NE, Pollak CT, Reiner K, Leshchiner E, Balazs EA. Hylan gel biomaterial: dermal and immunological compatibility. J Biomed Mater Res. 1993; 27:1129-34. https://doi.org/10.1002/jbm.820270903 PMid:8126011
Duranti F, Salti G, Bovani B, et al. Injectable hyaluronic acid gel for soft tissue augmentation. A clinical and histological study. Dermatol Surg. 1998; 24: 1317-25. https://doi.org/10.1111/j.1524-4725.1998.tb00007.x PMid:9865196
Varani J, Dame MK, Rittie L, et al. Decreased collagen production in chronologically aged skin. Am J Pathol. 2006; 168:1861-8. https://doi.org/10.2353/ajpath.2006.051302 PMid:16723701 PMCid:PMC1606623
Marmur ES, Phelps R, Goldberg DJ. Clinical, histologic and electron microscopic findings after injection of a calcium hydroxylapatite filler. J Cosmet Laser Ther. 2004; 6:223ï€6.
Berlin AL, Hussain M, Goldberg DJ. Calcium hydroxylapatite filler for facial rejuvenation: a histologic and immunohistochemical analysis. Dermatol Surg. 2008; 34(S1):64ï€7.
Mosman T.Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.J Immunol Methods. 1993; 65:55-63.https://doi.org/10.1016/0022-1759(83)90303-4
van Meerloo J, Kaspers GJ, Cloos J.Cell sensitivity assays: the MTT assay. Methods Mol Biol. 2011; 731:237-45. https://doi.org/10.1007/978-1-61779-080-5_20 PMid:21516412
Repetto G, del Peso A, Zuritsa JL. Neutral red uptake for the estimation of cell viability/cytotoxicity. Nat Prot. 2008; 3: 1125-31. https://doi.org/10.1038/nprot.2008.75 PMid:18600217
Knox, P, Uphill PF, Fry JR, Benford J, Balls M. The FRAME Multicentre project on In Vitro Toxicology.Chemical Toxicology. 1986; 24:457-63. https://doi.org/10.1016/0278-6915(86)90092-X
Wulf E, Deboben A, Faulstich H, Wieland T. Fluorescent phallotoxin, a tool for the visualization of cellular actin. Cell Biology. 1979; 4498: 1-15. https://doi.org/10.1073/pnas.76.9.4498
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