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European Journal of Medical Research
Published in: European Journal of Medical Research 1/2020

Open Access 01-12-2020 | Research

Dose- and time-dependent effects of hyaluronidase on structural cells and the extracellular matrix of the skin

Authors: Bettina Alexandra Buhren, Holger Schrumpf, Katharina Gorges, Oliver Reiners, Edwin Bölke, Jens W. Fischer, Bernhard Homey, Peter Arne Gerber

Published in: European Journal of Medical Research | Issue 1/2020

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Abstract

Introduction

Hyaluronic acid (hyaluronan; HA) is an essential component of the extracellular matrix (ECM) of the skin. The HA-degrading enzyme hyaluronidase (HYAL) is critically involved in the HA-metabolism. Yet, only little information is available regarding the skin’s HA–HYAL interactions on the molecular and cellular levels.

Objective

To analyze the dose- and time-dependent molecular and cellular effects of HYAL on structural cells and the HA-metabolism in the skin.

Materials and methods

Chip-based, genome-wide expression analyses (Affymetrix® GeneChip PrimeView™ Human Gene Expression Array), quantitative real-time PCR analyses, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (DAB), and in vitro wound healing assays were performed to assess dose-dependent and time-kinetic effects of HA and HYAL (bovine hyaluronidase, Hylase “Dessau”) on normal human dermal fibroblasts (NHDF), primary human keratinocytes in vitro and human skin samples ex vivo.

Results

Genome-wide expression analyses revealed an upregulation of HA synthases (HAS) up to 1.8-fold change in HA- and HYAL-treated NHDF. HA and HYAL significantly accelerated wound closure in an in vitro model for cutaneous wound healing. HYAL induced HAS1 and HAS2 mRNA gene expression in NHDF. Interestingly, low concentrations of HYAL (0.015 U/ml) resulted in a significantly higher induction of HAS compared to moderate (0.15 and 1.5 U/ml) and high concentrations (15 U/ml) of HYAL. This observation corresponded to increased concentrations of HA measured by ELISA in conditioned supernatants of HYAL-treated NHDF with the highest concentrations observed for 0.015 U/ml of HYAL. Finally, immunohistochemical analysis of human skin samples incubated with HYAL for up to 48 h ex vivo demonstrated that low concentrations of HYAL (0.015 U/ml) led to a pronounced accumulation of HA, whereas high concentrations of HYAL (15 U/ml) reduced dermal HA-levels.

Conclusion

HYAL is a bioactive enzyme that exerts multiple effects on the HA-metabolism as well as on the structural cells of the skin. Our results indicate that HYAL promotes wound healing and exerts a dose-dependent induction of HA-synthesis in structural cells of the skin. Herein, interestingly the most significant induction of HAS and HA were observed for the lowest concentration of HYAL.
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Literature
1.
Averbeck M, Gebhardt C, Anderegg U, Simon JC. Suppression of hyaluronan synthase 2 expression reflects the anthropogenic potential of glucocorticoids. Exp Dermatol. 2010;19:757–9.PubMedCrossRef
2.
Averbeck M, Gebhardt CA, Voigt S, Beilharz S, Anderegg U, Termeer CC, Sleeman JP, Simon JC. Differential regulation of hyaluronan metabolism in the epidermal and dermal compartments of human skin by UVB irradiation. J Invest Dermatol. 2007;127:687–97.PubMedCrossRef
3.
Aya KL, Stern R. Hyaluronan in wound healing: rediscovering a major player. Wound Repair Regen. 2014;22:579–93.PubMedCrossRef
4.
Barrientos S, Stojadinovic O, Golinko MS, Brem H, Tomic-Canic M. Growth factors and cytokines in wound healing. Wound Repair Regen. 2008;16:585–601.PubMedCrossRef
5.
Bissell MJ, Hall HG, Parry G. How does the extracellular matrix direct gene expression? J Theor Biol. 1982;99:31–68.PubMedCrossRef
6.
Buhren BA, Schrumpf H, Hoff NP, Bolke E, Hilton S, Gerber PA. Hyaluronidase: from clinical applications to molecular and cellular mechanisms. Eur J Med Res. 2016;21:5.PubMedPubMedCentralCrossRef
7.
Byl NN, Mckenzie A, West J, Stern M, Halliday B, Stern R, Wong T. Amniotic fluid enhances wound healing—a randomized controlled three week trial in mini Yucatan pigs. Eur J Phys Med Rehab. 1993;3:105–13.
8.
Camenisch TD, Schroeder JA, Bradley J, Klewer SE, Mcdonald JA. Heart-valve mesenchyme formation is dependent on hyaluronan-augmented activation of ErbB2-ErbB3 receptors. Nat Med. 2002;8:850–5.PubMedCrossRef
9.
Chen WY, Abatangelo G. Functions of hyaluronan in wound repair. Wound Repair Regen. 1999;7:79–89.PubMedCrossRef
10.
Ciccone V, Zazzetta M, Morbidelli L. Comparison of the effect of two hyaluronic acid preparations on fibroblast and endothelial cell functions related to angiogenesis. Cells. 2019;8:1479.PubMedCentralCrossRef
11.
Csoka AB, Frost GI, Stern R. The six hyaluronidase-like genes in the human and mouse genomes. Matrix Biol. 2001;20:499–508.PubMedCrossRef
12.
D’agostino A, Stellavato A, Busico T, Papa A, Tirino V, Papaccio G, La Gatta A, De Rosa M, Schiraldi C. In vitro analysis of the effects on wound healing of high- and low-molecular weight chains of hyaluronan and their hybrid H-HA/L-HA complexes. BMC Cell Biol. 2015;16:19.PubMedPubMedCentralCrossRef
13.
David-Raoudi M, Tranchepain F, Deschrevel B, Vincent JC, Bogdanowicz P, Boumediene K, Pujol JP. Differential effects of hyaluronan and its fragments on fibroblasts: relation to wound healing. Wound Repair Regen. 2008;16:274–87.PubMedCrossRef
14.
Delmage JM, Powars DR, Jaynes PK, Allerton SE. The selective suppression of immunogenicity by hyaluronic acid. Ann Clin Lab Sci. 1986;16:303–10.PubMed
15.
Dunphy JE, Udupa KN. Chemical and histochemical sequences in the normal healing of wounds. N Engl J Med. 1955;253:847–51.PubMedCrossRef
16.
17.
Flynn TC, Thompson DH, Hyun SH. Molecular weight analyses and enzymatic degradation profiles of the soft-tissue fillers Belotero Balance, Restylane, and Juvederm Ultra. Plast Reconstr Surg. 2013;132:22s–32s.PubMedCrossRef
18.
Fronza M, Caetano GF, Leite MN, Bitencourt CS, Paula-Silva FW, Andrade TA, Frade MA, Merfort I, Faccioli LH. Hyaluronidase modulates inflammatory response and accelerates the cutaneous wound healing. PLoS ONE. 2014;9:E112297.PubMedPubMedCentralCrossRef
19.
Funt D, Pavicic T. Dermal fillers in aesthetics: an overview of adverse events and treatment approaches. Clin Cosmet Investig Dermatol. 2013;6:295–316.PubMedPubMedCentral
20.
21.
Heldin P, Basu K, Olofsson B, Porsch H, Kozlova I, Kahata K. Deregulation of hyaluronan synthesis, degradation and binding promotes breast cancer. J Biochem. 2013;154:395–408.PubMedCrossRef
22.
Hirsch RJ, Brody HJ, Carruthers JD. Hyaluronidase in the office: a necessity for every dermasurgeon that injects hyaluronic acid. J Cosmet Laser Ther. 2007;9:182–5.PubMedCrossRef
23.
Homey B, Dieu-Nosjean MC, Wiesenborn A, Massacrier C, Pin JJ, Oldham E, Catron D, Buchanan ME, Muller A, Dewaal Malefyt R, Deng G, Orozco R, Ruzicka T, Lehmann P, Lebecque S, Caux C, Zlotnik A. Up-regulation of macrophage inflammatory protein-3 alpha/CCL20 and CC chemokine receptor 6 in psoriasis. J Immunol. 2000;164:6621–32.PubMedCrossRef
24.
Isnard N, Legeais JM, Renard G, Robert L. Effect of hyaluronan on MMP expression and activation. Cell Biol Int. 2001;25:735–9.PubMedCrossRef
25.
Jahn K, Homey B, Gerber PA. Management of complications after aesthetic hyaluronic acid injections. Hautarzt. 2014;65:851–3.PubMedCrossRef
26.
27.
Kaya G, Tran C, Sorg O, Hotz R, Grand D, Carraux P, Didierjean L, Stamenkovic I, Saurat JH. Hyaluronate fragments reverse skin atrophy by a CD44-dependent mechanism. Plos Med. 2006;3:E493.PubMedPubMedCentralCrossRef
28.
King SR, Hickerson WL, Proctor KG. Beneficial actions of exogenous hyaluronic acid on wound healing. Surgery. 1991;109:76–84.PubMed
29.
Larnier C, Kerneur C, Robert L, Moczar M. Effect of testicular hyaluronidase on hyaluronate synthesis by human skin fibroblasts in culture. Biochim Biophys Acta. 1989;1014:145–52.PubMedCrossRef
30.
Laurent TC, Fraser JR. The properties and turnover of hyaluronan. Ciba Found Symp. 1986;124:9–29.PubMed
31.
32.
Li L, Asteriou T, Bernert B, Heldin CH, Heldin P. Growth factor regulation of hyaluronan synthesis and degradation in human dermal fibroblasts: importance of hyaluronan for the mitogenic response of PDGF-BB. Biochem J. 2007;404:327–36.PubMedPubMedCentralCrossRef
33.
Lichtenberger BM, Gerber PA, Holcmann M, Buhren BA, Amberg N, Smolle V, Schrumpf H, Boelke E, Ansari P, Mackenzie C, Wollenberg A, Kislat A, Fischer JW, Rock K, Harder J, Schroder JM, Homey B, Sibilia M. Epidermal EGFR controls cutaneous host defense and prevents inflammation. Sci Transl Med. 2013;5:199111.CrossRef
34.
Meyer LJ, Stern R. Age-dependent changes of hyaluronan in human skin. J Invest Dermatol. 1994;102:385–9.PubMedCrossRef
35.
Mian N. Analysis of cell-growth-phase-related variations in hyaluronate synthase activity of isolated plasma-membrane fractions of cultured human skin fibroblasts. Biochem J. 1986a;237:333–42.PubMedPubMedCentralCrossRef
36.
Mian N. Characterization of a high-Mr plasma-membrane-bound protein and assessment of its role as a constituent of hyaluronate synthase complex. Biochem J. 1986b;237:343–57.PubMedPubMedCentralCrossRef
37.
Moczar M, Robert L. Stimulation of cell proliferation by hyaluronidase during in vitro aging of human skin fibroblasts. Exp Gerontol. 1993;28:59–68.PubMedCrossRef
38.
Monsuur HN, Boink MA, Weijers EM, Roffel S, Breetveld M, Gefen A, Van Den Broek LJ, Gibbs S. Methods to study differences in cell mobility during skin wound healing in vitro. J Biomech. 2016;49:1381–7.PubMedCrossRef
39.
40.
Narayanan R, Kuppermann BD. Hyaluronidase for pharmacologic vitreolysis. Dev Ophthalmol. 2009;44:20–5.PubMedCrossRef
41.
Noble PW. Hyaluronan and its catabolic products in tissue injury and repair. Matrix Biol. 2002;21:25–9.PubMedCrossRef
42.
Philipson LH, Schwartz NB. Subcellular localization of hyaluronate synthetase in oligodendroglioma cells. J Biol Chem. 1984;259:5017–23.PubMed
43.
Philipson LH, Westley J, Schwartz NB. Effect of hyaluronidase treatment of intact cells on hyaluronate synthetase activity. Biochemistry. 1985;24:7899–906.CrossRef
44.
45.
Quan T, Wang F, Shao Y, Rittie L, Xia W, Orringer JS, Voorhees JJ, Fisher GJ. Enhancing structural support of the dermal microenvironment activates fibroblasts, endothelial cells, and keratinocytes in aged human skin in vivo. J Invest Dermatol. 2013;133:658–67.PubMedCrossRef
46.
Reed MJ, Damodarasamy M, Chan CK, Johnson MN, Wight TN, Vernon RB. Cleavage of hyaluronan is impaired in aged dermal wounds. Matrix Biol. 2013;32:45–51.PubMedCrossRef
47.
Rock K, Grandoch M, Majora M, Krutmann J, Fischer JW. Collagen fragments inhibit hyaluronan synthesis in skin fibroblasts in response to ultraviolet B (UVB): new insights into mechanisms of matrix remodeling. J Biol Chem. 2011;286:18268–76.PubMedPubMedCentralCrossRef
48.
49.
Stern R. Devising a pathway for hyaluronan catabolism: are we there yet? Glycobiology. 2003;13:105r–15r.PubMedCrossRef
50.
51.
52.
Stern R, Maibach HI. Hyaluronan in skin: aspects of aging and its pharmacologic modulation. Clin Dermatol. 2008;26:106–22.PubMedCrossRef
53.
Stern R, Asari AA, Sugahara KN. Hyaluronan fragments: an information-rich system. Eur J Cell Biol. 2006;85:699–715.PubMedCrossRef
54.
Takahashi Y, Li L, Kamiryo M, Asteriou T, Moustakas A, Yamashita H, Heldin P. Hyaluronan fragments induce endothelial cell differentiation in a CD44- and CXCL1/GRO1-dependent manner. J Biol Chem. 2005;280:24195–204.PubMedCrossRef
55.
Tian X, Azpurua J, Hine C, Vaidya A, Myakishev-Rempel M, Ablaeva J, Mao Z, Nevo E, Gorbunova V, Seluanov A. High-molecular-mass hyaluronan mediates the cancer resistance of the naked mole rat. Nature. 2013;499:346–9.PubMedPubMedCentralCrossRef
56.
Wahby A, Dicaprio KD, Stern R. Hyaluronan inside and outside of skin. In: Lodén M, Maibach H, editors. Treatment of dry skin syndrome. Berlin: Springer; 2012.
57.
Wang F, Garza LA, Kang S, Varani J, Orringer JS, Fisher GJ, Voorhees JJ. In vivo stimulation of de novo collagen production caused by cross-linked hyaluronic acid dermal filler injections in photodamaged human skin. Arch Dermatol. 2007;143:155–63.PubMed
58.
Weber GC, Buhren BA, Schrumpf H, Wohlrab J, Gerber PA. Clinical applications of hyaluronidase. Adv Exp Med Biol. 2019;1148:255–77.PubMedCrossRef
59.
West DC, Hampson IN, Arnold F, Kumar S. Angiogenesis induced by degradation products of hyaluronic acid. Science. 1985;228:1324–6.PubMedCrossRef
60.
Wohlrab J, Finke R, Franke WG, Wohlrab A. Clinical trial for safety evaluation of hyaluronidase as diffusion enhancing adjuvant for infiltration analgesia of skin with lidocaine. Dermatol Surg. 2012;38:91–6.PubMedCrossRef
Metadata
Title
Dose- and time-dependent effects of hyaluronidase on structural cells and the extracellular matrix of the skin
Authors
Bettina Alexandra Buhren
Holger Schrumpf
Katharina Gorges
Oliver Reiners
Edwin Bölke
Jens W. Fischer
Bernhard Homey
Peter Arne Gerber
Publication date
01-12-2020
Publisher
BioMed Central
Published in
European Journal of Medical Research / Issue 1/2020
Electronic ISSN: 2047-783X
DOI
https://doi.org/10.1186/s40001-020-00460-z

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