Drug Delivery and Translational Research

, Volume 8, Issue 5, pp 1014–1024 | Cite as

Comparison of healing of full-thickness skin wounds grafted with multidirectional or unidirectional autologous artificial dermis: differential delivery of healing biomarkers

  • M. R. FontanillaEmail author
  • S. Casadiegos
  • R.H. Bustos
  • M.A. Patarroyo
Original Article


Cytokines, chemokines, and growth and remodeling factors orchestrate wound healing when skin damage occurs. During early stages, when the wound is still open, detection and quantification of these compounds might provide biomarkers of skin wound healing, which could aid to complete the scenario provided by clinical follow-up data and histological and histomorphometric analyses. This work assessed and compared the healing of full-thickness skin wounds grafted with artificial dermis made with autologous skin fibroblasts and unidirectional or multidirectional type I collagen scaffolds to test this hypothesis. Biomarkers of healing were detected and quantified in the culture medium of artificial dermis and exudates from the grafted wounds. Clinical follow-up of animals and histological and histomorphometric analysis showed differences in graft integration, wound closure, and histological and histomorphometric parameters. Surface plasmon resonance quantification of 13 healing biomarkers indicated differential secretion of most of the quantified factors in culture medium by the multidirectional and unidirectional artificial dermis. Also, there were significant differences between the concentration of some of the factors analyzed in the exudates of wounds grafted with the evaluated artificial dermis. These findings suggest that differential delivery of healing biomarkers induced by the directionality of the scaffold used to produce the multidirectional and unidirectional dermis was sufficient to create two skin wound microenvironments that determined a different outcome of healing. Overall, data indicate that healing of wounds grafted with multidirectional autologous artificial dermis is better than that of the wounds grafted with the unidirectional one.


Multidirectional and unidirectional autologous artificial dermis Differential delivery of bioactive compounds Fiber orientation Healing of full-thickness skin wounds Healing biomarkers SPR quantification 


Authors’ contributions

M. R. Fontanilla wrote the manuscript and participated in the planning of the experiments and discussion of results. S. Casadiegos carried out most of the experimental work, prepared all the figures of the manuscript, participated in the planning of the experiments and discussion of results, and helped in the preparation of the manuscript. R.H. Bustos obtained the data for the calibration curves, participated in the planning of the SPR-experiments, discussion of SPR-results, and critical revision of the manuscript. M.A. Patarroyo participated in the discussion of animal experiment results and critical revision of the manuscript.

Funding information

This work was supported by the Colombian Administrative Department of Science, Technology, and Innovation (Colciencias) Grant 1101-569-35037. Casadiegos S. and Bustos R.H. were supported by the same grant. Casadiegos S. was also supported by a Colciencias-Colfuturo Ph.D. Scholarship to finish his Ph.D. thesis work. Authors would like to thank Miguelangel Moncayo Donoso for helping with the edition of the figures.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13346_2018_528_MOESM1_ESM.pdf (8.2 mb)
ESM 1 (PDF 8440 kb)


  1. 1.
    Enoch S, Leaper DJ. Basic science of wound healing. Surg. 2005;23(2):37–42.Google Scholar
  2. 2.
    Yannas IV, Kwan MD, Longaker MT. Early fetal healing as a model for adult organ regeneration. Tissue Eng. 2007;13(8):1789–98.CrossRefPubMedGoogle Scholar
  3. 3.
    Yannas IV, Lee E, Orgill DP, Skrabut EM, Murphy GF. Synthesis and characterization of a model extracellular matrix that induces partial regeneration of adult mammalian skin. Proc Natl Acad Sci U S A. 1989;86(3):933–7.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453(7193):314–21.CrossRefPubMedGoogle Scholar
  5. 5.
    Krafts KP. Tissue repair. Organ. 2010;6(4):225–33.Google Scholar
  6. 6.
    Barrientos S, Stojadinovic O, Golinko MS, Brem H, Tomic-Canic M. Growth factors and cytokines in wound healing. Wound Repair Regen. 2008;16(5):585–601.CrossRefPubMedGoogle Scholar
  7. 7.
    Mast BA, Schultz GS. Interactions of cytokines, growth factors, and proteases in acute and chronic wounds. Wound Repair Regen. 1996;4(4):411–20.CrossRefPubMedGoogle Scholar
  8. 8.
    Werner S, Grose R. Regulation of wound healing by growth factors and cytokines. Physiol Rev. 2003;83(3):835–70.CrossRefPubMedGoogle Scholar
  9. 9.
    Mooney DP, O’Reilly M, Gamelli RL. Tumor necrosis factor and wound healing. Ann Surg. 1990;211(2):124–9.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Salmon-Ehr V, Ramont L, Godeau G, Birembaut P, Guenounou M, Bernard P, et al. Implication of interleukin-4 in wound healing. Lab Investig. 2000;80(8):1337–43.CrossRefPubMedGoogle Scholar
  11. 11.
    Maxson S, Lopez EA, Yoo D, Danilkovitch-Miagkova A, LeRoux MA. Concise review: role of mesenchymal stem cells in wound repair. Stem Cells Transl Med. 2012;1(2):142–9.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Bao P, Kodra A, Tomic-Canic M, Golinko MS, Ehrlich HP, Brem H. The role of vascular endothelial growth factor in wound healing. J Surg Res. 2009;153(2):347–58.CrossRefPubMedGoogle Scholar
  13. 13.
    Kämpfer H, Pfeilschifter J, Frank S. Expressional regulation of angiopoietin-1 and -2 and the tie-1 and -2 receptor tyrosine kinases during cutaneous wound healing: a comparative study of normal and impaired repair. Lab Investig. 2001;81(3):361–73.CrossRefPubMedGoogle Scholar
  14. 14.
    Bloch W. The angiogenesis inhibitor endostatin impairs blood vessel maturation during wound healing. FASEB J. 2000;14(15):2373–6.CrossRefPubMedGoogle Scholar
  15. 15.
    Espinosa L, Sosnik A, Fontanilla MR. Development and preclinical evaluation of acellular collagen scaffolding and autologous artificial connective tissue in the regeneration of oral mucosa wounds. Tissue Eng Part A. 2010;16(5):1667–79.CrossRefPubMedGoogle Scholar
  16. 16.
    Fontanilla MR, Espinosa LG. In vitro and in vivo assessment of oral autologous artificial connective tissue characteristics that influence its performance as a graft. Tissue Eng Part A. 2012;18(17–18):1857–66.CrossRefPubMedGoogle Scholar
  17. 17.
    Bustos RH, Suesca E, Millán D, González JM, Fontanilla MR. Real-time quantification of proteins secreted by artificial connective tissue made from uni- or multidirectional collagen I scaffolds and oral mucosa fibroblasts. Anal Chem. 2014;86(5):2421–8.CrossRefPubMedGoogle Scholar
  18. 18.
    Nguyen H, Park J, Kang S, Kim M. Surface plasmon resonance: a versatile technique for biosensor applications. Sensors. 2015;15(5):10481–510.CrossRefPubMedGoogle Scholar
  19. 19.
    Chou T-H, Chuang C-Y, Wu C-M. Quantification of interleukin-6 in cell culture medium using surface plasmon resonance biosensors. Cytokine. 2010;51(1):107–11.CrossRefPubMedGoogle Scholar
  20. 20.
    Martinez-Perdiguero J, Retolaza A, Bujanda L, Merino S. Surface plasmon resonance immunoassay for the detection of the TNFα biomarker in human serum. Talanta. 2014;119:492–7.CrossRefPubMedGoogle Scholar
  21. 21.
    Ulman A. Formation and structure of self-assembled monolayers. Chem Rev. 1996;96(4):1533–54.CrossRefPubMedGoogle Scholar
  22. 22.
    Camillone N. Diffusion-limited thiol adsorption on the gold(111) surface. Langmuir. 2004;20(4):1199–206.CrossRefPubMedGoogle Scholar
  23. 23.
    Battaglia TM, Masson J-F, Sierks MR, Beaudoin SP, Rogers J, Foster KN, et al. Quantification of cytokines involved in wound healing using surface plasmon resonance. Anal Chem. 2005;77(21):7016–23.CrossRefPubMedGoogle Scholar
  24. 24.
    Snowden JM, Kennedy DF, Cliff WJ. Wound contraction. The effects of scab formation and the nature of the wound bed. Aust J Exp Biol Med Sci. 1982;60(Pt 1):73–82.CrossRefPubMedGoogle Scholar
  25. 25.
    Niksa L, Geneviève M, Reyes-Gomez E, Lilin T, Crosaz O, Dohan Ehrenfest DM. Cutaneous reepithelialization and wound contraction after skin biopsies in rabbits: a mathematical model for healing and remodelling index. Vet Arh. 2010;80(5):637–52.Google Scholar
  26. 26.
    Suesca E, Dias AMA, Braga MEM, de Sousa HC, Fontanilla MR. Multifactor analysis on the effect of collagen concentration, cross-linking and fiber/pore orientation on chemical, microstructural, mechanical and biological properties of collagen type I scaffolds. Mater Sci Eng C. 2017;77:333–41.CrossRefGoogle Scholar
  27. 27.
    Lohana P, Hassan S, Watson S. Integra™ in burns reconstruction: our experience and report of an unusual immunological reaction. Ann Burns Fire Disasters. 2014;27(1):17–21.PubMedPubMedCentralGoogle Scholar
  28. 28.
    Veves A, Falanga V, Armstrong DG, Sabolinski ML. Graftskin, a human skin equivalent, is effective in the management of noninfected neuropathic diabetic foot ulcers: a prospective randomized multicenter clinical trial. Diabetes Care. 2001;24(2):290–5.CrossRefPubMedGoogle Scholar
  29. 29.
    Falanga V, Sabolinski M. A bilayered living skin construct (APLIGRAF) accelerates complete closure of hard-to-heal venous ulcers. Wound Repair Regen. 1999;7(4):201–7.CrossRefPubMedGoogle Scholar
  30. 30.
    Hull BE, Sher SE, Rosen S, Church D, Bell E. Fibroblasts in isogeneic skin equivalents persist for long periods after grafting. J Invest Dermatol. 1983;81(5):436–8.CrossRefPubMedGoogle Scholar
  31. 31.
    Bell E, Ehrlich HP, Buttle DJ, Nakatsuji T. Living tissue formed in vitro and accepted as skin-equivalent tissue of full thickness. Science. 1981;211(4486):1052–4.CrossRefPubMedGoogle Scholar
  32. 32.
    Burke J, Yannas I, Quinby Jr W, Bondoc C, Jung W. Successful use of a physiologically acceptable artificial skin in the treatment of extensive burn injury. Ann Surg. 1981;194(4):413–28.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Lee LF, Porch JV, Spenler W, Garner WL. Integra in lower extremity reconstruction after burn injury. Plast Reconstr Surg. 2008;121(4):1256–62.CrossRefPubMedGoogle Scholar
  34. 34.
    Ghahary A, Marcoux Y, Karimi-Busheri F, Li Y, Tredget EE, Kilani RT, et al. Differentiated keratinocyte-releasable stratifin (14-3-3 Sigma) stimulates MMP-1 expression in dermal fibroblasts. J Invest Dermatol. 2005;124(1):170–7.CrossRefPubMedGoogle Scholar
  35. 35.
    Skalli O, Giulio G. The biology of the myofibroblast relationship to wound contraction and fibrocontractive diseases. In: Clark RAF, Henson PM, editors. The molecular and cellular biology of wound repair. Boston, MA: Springer US; 1988. p. 373–402.CrossRefGoogle Scholar
  36. 36.
    Millán D, Jiménez RA, Nieto LE, Linero I, Laverde M, Fontanilla MR. Preclinical evaluation of collagen type I scaffolds, including gelatin-collagen microparticles and loaded with a hydroglycolic calendula officinalis extract in a lagomorph model of full-thickness skin wound. Drug Deliv Transl Res. 2016;6(1):57–66.CrossRefPubMedGoogle Scholar
  37. 37.
    Galiano RD, Michaels VJ, Dobryansky M, Levine JP, Gurtner GC. Quantitative and reproducible murine model of excisional wound healing. Wound Repair Regen. 2004;12(4):485–92.CrossRefPubMedGoogle Scholar
  38. 38.
    Rittié L. Cellular mechanisms of skin repair in humans and other mammals. J Cell Commun Signal. 2016 Jun 12;10(2):103–20.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Colangelo E, Comenge J, Paramelle D, Volk M, Chen Q, Lévy R. Characterizing self-assembled monolayers on gold nanoparticles. Bioconjug Chem. 2017;28(1):11–22.CrossRefPubMedGoogle Scholar
  40. 40.
    Matharu Z, Bandodkar AJ, Gupta V, Malhotra BD. Fundamentals and application of ordered molecular assemblies to affinity biosensing. Chem Soc Rev. 2012;41(3):1363–402.CrossRefPubMedGoogle Scholar

Copyright information

© Controlled Release Society 2018

Authors and Affiliations

  • M. R. Fontanilla
    • 1
    Email author
  • S. Casadiegos
    • 1
  • R.H. Bustos
    • 1
  • M.A. Patarroyo
    • 2
    • 3
  1. 1.Grupo de Trabajo en Ingeniería de Tejidos, Departamento de FarmaciaUniversidad Nacional de ColombiaBogotáColombia
  2. 2.Molecular Biology and Immunology DepartmentFundación Instituto de Inmunología de Colombia (FIDIC)BogotáColombia
  3. 3.Basic Sciences DepartmentUniversidad del RosarioBogotáColombia

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