Skip to main content
Log in

The reconstructed skin model as a new tool for investigating in vitro dermal fillers: increased fibroblast activity by hyaluronic acid

  • Investigative Report
  • Published:
European Journal of Dermatology

Abstract

Background

Clinical studies on dermal fillers have essentially focused upon visible improvement of skin quality and any eventual side effects, whereas very little is known about their detailed biological effects.

Objectives

New skin equivalent models were created to investigate the biological impact of hyaluronic acid (HA) fillers on the dermal compartment in vitro.

Materials and methods

Two different reconstructed skin models were developed to incorporate HA within the collagen fibers. In the mixed model, HA was distributed throughout the whole collagen gel whereas the HA was concentrated in the center of collagen gel in the inclusion model.

Results

A comparison of the addition of fillers in two models of reconstructed skin has permitted a better understanding of the biological impact of HA fillers. Protein profiling of supernatants from both models suggested a regulation of MMP-1 secretion by fibroblasts as a function of HA volume, distribution in the dermis and degree of cross-linking. Immunostaining of the inclusion model revealed increased production of type I and III procollagens close to the cross-linked HA. Fibroblasts located in this area showed a fusiform morphology as well as an increase in α-smooth actin expression. The observed increase in collagen production may thus result in part from tension in fibroblasts surrounding the cross-linked HA.

Conclusion

The inclusion reconstructed skin model, as compared to the mixed model, presented here, appears to be a useful tool for investigating the properties of various fillers in vitro and closer to the in vivo situation; our results show that HA fillers promote in vitro remodeling of the dermis by fibroblasts.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Buck DW, Alam M, Kim JY. Injectable fillers for facial rejuvenation: a review. J Plast Reconstr Aesthet Surg 2009; 62: 11–8.

    Article  PubMed  Google Scholar 

  2. Monheit GD, Coleman KM. Hyaluronic acid fillers. Dermatol Ther 2006; 19: 141–50.

    Article  PubMed  Google Scholar 

  3. Stern R, Maibach HI. Hyaluronan in skin: aspects of aging and its pharmacologic modulation. Clin Dermatol 2008; 26: 106–22.

    Article  PubMed  Google Scholar 

  4. Tammi RH, Tammi MI. Hyaluronan accumulation in wounded epidermis: a mediator of keratinocyte activation. J Invest Dermatol 2009; 129: 1858–60.

    Article  CAS  PubMed  Google Scholar 

  5. Noble PW. Hyaluronan and its catabolic products in tissue injury and repair. Matrix Biol 2002; 21: 25–9.

    Article  CAS  PubMed  Google Scholar 

  6. Narins RS, Brandt F, Leyden J, Lorenc ZP, Rubin M, Smith S. A randomized, double-blind, multicenter comparison of the efficacy and tolerability of Restylane versus Zyplast for the correction of nasolabial folds. Dermatol Surg 2003; 29: 588–95.

    PubMed  Google Scholar 

  7. Bogdan A I A, Baumann L. Hyaluronic acid gel (Juvederm) preparations in the treatment of facial wrinkles and folds. Clin Interv Aging 2008; 3: 629–34.

    Google Scholar 

  8. Carruthers A, Carey W, De LC, Remington K, Schachter D, Sapra S. Randomized, double-blind comparison of the efficacy of two hyaluronic acid derivatives, restylane perlane and hylaform, in the treatment of nasolabial folds. Dermatol Surg 2005; 31: 1591–8.

    Article  CAS  PubMed  Google Scholar 

  9. Fernandez-Cossio S, Castano-Oreja MT. Biocompatibility of two novel dermal fillers: histological evaluation of implants of a hyaluronic acid filler and a polyacrylamide filler. Plast Reconstr Surg 2006; 117: 1789–96.

    Article  CAS  PubMed  Google Scholar 

  10. Arlette JP, Trotter MJ. Anatomic location of hyaluronic acid filler material injected into nasolabial fold: a histologic study. Dermatol Surg 2008; 34(Suppl 1): S56–62.

    CAS  PubMed  Google Scholar 

  11. Kerscher M, Bayrhammer J, Reuther T. Rejuvenating influence of a stabilized hyaluronic acid-based gel of nonanimal origin on facial skin aging. Dermatol Surg 2008; 34: 720–6.

    CAS  PubMed  Google Scholar 

  12. Turlier V, Rouquier A, Black D, et al. Assessment of the clinical efficacy of a hyaluronic acid-based deep wrinkle filler using new instrumental methods. J Cosmet Laser Ther 2010; 12: 195–202.

    Article  PubMed  Google Scholar 

  13. Wang F, Garza LA, Kang S, et al. 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.

    CAS  PubMed  Google Scholar 

  14. Mine S, Fortunel NO, Pageon H, Asselineau D. Aging alters functionally human dermal papillary fibroblasts but not reticular fibroblasts: a new view of skin morphogenesis and aging. PLoS ONE 2008; 3: e4066.

    Article  PubMed Central  PubMed  Google Scholar 

  15. Rheinwald JG, Green H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 1975; 6: 331–43.

    Article  CAS  PubMed  Google Scholar 

  16. Asselineau D, Prunieras M. Reconstruction of’ simplified’ skin: control of fabrication. Br J Dermatol 1984; 111(Suppl 27): 219–22.

    Article  PubMed  Google Scholar 

  17. Bell E, Sher S, Hull B, et al. The reconstitution of living skin. J Invest Dermatol 1983; 81: 2s–10s.

    Article  CAS  PubMed  Google Scholar 

  18. Mehra TD, Ghosh K, Shu XZ, Prestwich GD, Clark RA. Molecular stenting with a crosslinked hyaluronan derivative inhibits collagen gel contraction. J Invest Dermatol 2006; 126: 2202–9.

    Article  CAS  PubMed  Google Scholar 

  19. Canty EG, Kadler KE. Procollagen trafficking, processing and fibrillogenesis. J Cell Sc 2005; 118: 1341–53.

    Article  CAS  Google Scholar 

  20. Eckes B, Zweers MC, Zhang ZG, et al. Mechanical tension and integrin alpha 2 beta 1 regulate fibroblast functions. J Investig Dermatol Symp Proc 2006; 11: 66–72.

    Article  CAS  PubMed  Google Scholar 

  21. Lambert CA, Soudant EP, Nusgens BV, Lapiere CM. Pretranslational regulation of extracellular matrix macromolecules and collagenase expression in fibroblasts by mechanical forces. Lab Invest 1992; 66: 444–51.

    CAS  PubMed  Google Scholar 

  22. Parsons M, Kessler E, Laurent GJ, Brown RA, Bishop JE. Mechanical load enhances procollagen processing in dermal fibroblasts by regulating levels of procollagen C-proteinase. Exp Cell Res 1999; 252: 319–31.

    Article  CAS  PubMed  Google Scholar 

  23. Kessler D, Dethlefsen S, Haase I, et al. Fibroblasts in mechanically stressed collagen lattices assume a “synthetic” phenotype. J Biol Chem 2001; 276: 36575–85.

    Article  CAS  PubMed  Google Scholar 

  24. Lambert CA, Colige AC, Lapiere CM, Nusgens BV. Coordinated regulation of procollagens I and III and their post-translational enzymes by dissipation of mechanical tension in human dermal fibroblasts. Eur J Cell Biol 2001; 80: 479–85.

    Article  CAS  PubMed  Google Scholar 

  25. Varani J, Dame MK, Rittie L, et al. Decreased collagen production in chronologically aged skin: roles of age-dependent alteration in fibroblast function and defective mechanical stimulation. Am J Pathol 2006; 168: 1861–8.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Varani J, Spearman D, Perone P, et al. Inhibition of type I procollagen synthesis by damaged collagen in photoaged skin and by collagenase-degraded collagen in vitro. Am J Pathol 2001; 158: 931–42.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Varani J, Perone P, Fligiel SE, Fisher GJ, Voorhees JJ. Inhibition of type I procollagen production in photodamage: correlation between presence of high molecular weight collagen fragments and reduced procollagen synthesis. J Invest Dermatol 2002; 119: 122–9.

    Article  CAS  PubMed  Google Scholar 

  28. Dumas M, Chaudagne C, Bonte F, Meybeck A. In vitro biosynthesis of type I and III collagens by human dermal fibroblasts from donors of increasing age. Mech Ageing Dev 1994; 73: 179–87.

    Article  CAS  PubMed  Google Scholar 

  29. Reed MJ, Ferara NS, Vernon RB. Impaired migration, integrin function, and actin cytoskeletal organization in dermal fibroblasts from a subset of aged human donors. Mech Ageing Dev 2001; 122: 1203–20.

    Article  CAS  PubMed  Google Scholar 

  30. Kablik J, Monheit GD, Yu L, Chang G, Gershkovich J. Comparative physical properties of hyaluronic acid dermal fillers. Dermatol Surg 2009; 35(Suppl 1): 302–12.

    Article  CAS  PubMed  Google Scholar 

  31. Tezel A, Fredrickson GH. The science of hyaluronic acid dermal fillers. J Cosmet Laser Ther 2008; 10: 35–42.

    Article  PubMed  Google Scholar 

  32. Park KY, Kim HK, Kim BJ. Comparative study of hyaluronic acid fillers by in vitro and in vivo testing. J Eur Acad Dermatol Venereol 2014; 28: 565–8.

    Article  CAS  PubMed  Google Scholar 

  33. Fagot D, Asselineau D, Bernerd F. Matrix metalloproteinase-1 production observed after solar-simulated radiation exposure is assumed by dermal fibroblasts but involves a paracrine activation through epidermal keratinocytes. Photochem Photobiol 2004; 79: 499–505.

    Article  CAS  PubMed  Google Scholar 

  34. Turlier V, Delalleau A, Casas C, et al. Association between collagen production and mechanical stretching in dermal extracellular matrix: In vivo effect of cross-linked hyaluronic acid filler. A randomised, placebo-controlled study. J Dermatol Sci 2013; 69: 187–94.

    Article  CAS  PubMed  Google Scholar 

  35. Quan T, Wang F, Shao Y, et al. 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.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  36. Yang C, Cao M, Liu H, et al. The high and low molecular weight forms of hyaluronan have distinct effects on CD44 clustering. JBC 2012; 287: 43094–107.

    Article  CAS  Google Scholar 

  37. Lisignoli G, Cristino S, Piacentini A, Cavallo C, Caplan AI, Facchini A. Hyaluronan-based polymer scaffold modulates the expression of inflammatory and degradative factors in mesenchymal stem cells: Involvement of Cd44 and Cd54. J Cell Physiol 2006; 207: 364–73.

    Article  CAS  PubMed  Google Scholar 

  38. Isnard N, Legeais JM, Renard G, Robert L. Effect of hyaluronan on MMP expression and activation. Cell Biol Int 2001; 25: 735–9.

    Article  CAS  PubMed  Google Scholar 

  39. Isnard N, Robert L, Renard G. Effect of sulfated GAGs on the expression and activation of MMP-2 and MMP-9 in corneal and dermal explant cultures. Cell Biol Int 2003; 27: 779–84.

    Article  CAS  PubMed  Google Scholar 

  40. Huang L, Gu H, Burd A. A reappraisal of the biological effects of hyaluronan on human dermal fibroblast. J Biomed Mater Res A 2009; 90: 1177–85.

    Article  PubMed  Google Scholar 

  41. David-Raoudi M, Tranchepain F, Deschrevel B, et al. Differential effects of hyaluronan and its fragments on fibroblasts: relation to wound healing. Wound Repair Regen 2008; 16: 274–87.

    Article  PubMed  Google Scholar 

  42. Gao F, Liu Y, He Y, et al. Hyaluronan oligosaccharides promote excisional wound healing through enhanced angiogenesis. Matrix Biol 2010; 29: 107–16.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sarah Girardeau-Hubert.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Girardeau-Hubert, S., Teluob, S., Pageon, H. et al. The reconstructed skin model as a new tool for investigating in vitro dermal fillers: increased fibroblast activity by hyaluronic acid. Eur J Dermatol 25, 312–322 (2015). https://doi.org/10.1684/ejd.2015.2563

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1684/ejd.2015.2563

Keywords

Navigation