Drug Delivery and Translational Research

, Volume 9, Issue 1, pp 25–36 | Cite as

Evaluation of collagen type I scaffolds including gelatin-collagen microparticles and Aloe vera in a model of full-thickness skin wound

  • Liliana Gil-Cifuentes
  • Ronald A. Jiménez
  • Marta R. FontanillaEmail author
Original Article


Research on collagen type I scaffolds with Aloe vera is sparse. The aim of this work was to develop collagen type I scaffolds with gelatin-collagen microparticles and loaded with a dispersion of A. vera, to assess their performance as grafting material for healing of skin wounds. Scaffolds were evaluated in a Cavia porcellus model with full-thickness skin wound and compared with wounds healed by secondary intention (controls). Animals grafted with scaffolds without A. vera and their control wounds were also included in the study. Evaluation of enzymatic degradation and percentage of the scaffolds’ free amino groups—as an indirect assessment of their cross-linking—were also carried out because A. vera contains compounds which affect their stability. We found that dispersions of lyophilized A. vera extract loaded on scaffolds do not have cytotoxic potential, and they decrease collagenase degradation of scaffolds in the range of 0.1 to 0.3% w/v in a dose-dependent manner. Only the A. vera dispersion with the highest concentration (0.3% w/v) decreased the percentage of free amino groups, which are the ones involved in the cross-link of collagen fibers. This finding suggests that cross-linking is not the mechanism by which the tested dispersions stabilize the scaffolds. Preclinical, histochemical, and histomorphometric analyses of repaired wound tissue indicate that loading collagen type I scaffolds, including microparticles of gelatin-collagen, with A. vera in the concentrations tested does not improve wound healing. Low biodegradability of the tested scaffolds caused by the inhibition of collagenase activity might account for these results.


Aloe vera Collagen type I scaffolds Microparticles of gelatin-collagen Collagenase degradation Cross-linking Wound healing Total-thickness skin wounds 



The authors would like to thank Dr. Felipe Cabello for the critical reading of the manuscript and his suggestions for improving it, Ph.D. candidate Julia Morales and Dr. Manuel Morales for editing the English, Ph.D. candidate Diana Millán for helping with the edition of the figures, and DVM Alejandra Muñoz for helping with the animal experiments.

Financial information

This work was supported by the Colombian Administrative Department of Science, Technology and Innovation (Colciencias) Grant 1101-569-34982.

Compliance of ethical standards

All institutional and national guidelines for the care and use of laboratory animal were followed.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Wells A, Nuschke A, Yates CC. Skin tissue repair: matrix microenvironmental influences. Matrix Biol. 2016;49:25–36.CrossRefGoogle Scholar
  2. 2.
    Chattopadhyay S, Raine RT, Glick GD. Review collagen-based biomaterials for wound healing. Biopolymers. 2014;101:821–33.CrossRefGoogle Scholar
  3. 3.
    Zhang Q, Lu H, Kawazoe N, Chen G. Preparation of collagen scaffolds with controlled pore structures and improved mechanical property for cartilage tissue engineering. J Bioact Compat Polym. 2013;28:426–38.CrossRefGoogle Scholar
  4. 4.
    Soller EC, Tzeranis DS, Miu K, So PTC, Yannas IV. Common features of optimal collagen scaffolds that disrupt wound contraction and enhance regeneration both in peripheral nerves and in skin. Biomaterials. 2012;33:4783–91.CrossRefGoogle Scholar
  5. 5.
    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 A. 2010;16:1667–79.CrossRefGoogle Scholar
  6. 6.
    Jimenez RA, Millan D, Suesca E, Sosnik A, Fontanilla MR. Controlled release of an extract of Calendula officinalis flowers from a system based on the incorporation of gelatin-collagen microparticles into collagen I scaffolds: design and in vitro performance. Drug Deliv Transl Res. 2015;5:209–18.CrossRefGoogle Scholar
  7. 7.
    Hamman JH. Composition and applications of Aloe vera leaf gel. Molecules. 2008;13:1599–616.CrossRefGoogle Scholar
  8. 8.
    Maenthaisong R, Chaiyakunapruk N, Niruntraporn S, Kongkaew C. The efficacy of Aloe vera used for burn wound healing: a systematic review. Burns. 2007;33:713–8.CrossRefGoogle Scholar
  9. 9.
    Oryan A, Mohammadalipour A, Moshiri A, Tabandeh MR. Topical application of Aloe vera accelerated wound healing, modeling, and remodeling: an experimental study with significant clinical value. Ann Plast Surg. 2016;77:37–46.CrossRefGoogle Scholar
  10. 10.
    Gupta VK, Malhotra S. Pharmacological attribute of Aloe vera: revalidation through experimental and clinical studies. Ayu. 2012;33:193–6.CrossRefGoogle Scholar
  11. 11.
    Choi S, Son B, Son Y, Park Y, Lee S, Chung M. The wound-healing effect of a glycoprotein fraction isolated from Aloe vera. Br J Dermatol. 2001;145:535–45.CrossRefGoogle Scholar
  12. 12.
    Lee M, Lee O, Yoon S, Lee S, Chung M, Park Y, et al. In vitro angiogenic activity of Aloe vera gel on calf pulmonary artery endothelial (CPAE) cells. Arch Pharm Res. 1998;21:260–5.CrossRefGoogle Scholar
  13. 13.
    Jettanacheawchankit S, Sasithanasate S, Sangvanich P, Banlunara W, Thunyakitpisal P. Acemannan stimulates gingival fibroblast proliferation; expressions of keratinocyte growth factor-1, vascular endothelial growth factor, and type I collagen; and wound healing. J Pharmacol Sci. 2009;109:525–31.CrossRefGoogle Scholar
  14. 14.
    Chithra P, Sajithlal GB, Chandrakasan G. Influence of Aloe vera on the healing of dermal wounds in diabetic rats. J Ethnopharmacol. 1998;59:195–201.CrossRefGoogle Scholar
  15. 15.
    Chung M-H, Choi S. A review on the relationship between Aloe vera components and their biologic effects. Semin Integr Med. 2003;1:53–62.CrossRefGoogle Scholar
  16. 16.
    Wahedi HM, Jeong M, Chae JK, Do SG, Yoon H, Kim SY. Aloesin from Aloe vera accelerates skin wound healing by modulating MAPK/Rho and Smad signaling pathways in vitro and in vivo. Phytomedicine. 2017;28:19–26.CrossRefGoogle Scholar
  17. 17.
    Salehi M, Farzamfar S, Bastami F, Tajerian R. Fabrication and characterization of electrospun PLLA/collagen nanofibrous scaffold coated with chitosan to sustain release of Aloe vera gel for skin tissue engineering. Biomed Eng Appl Basis Commun. 2016;28:1650035.CrossRefGoogle Scholar
  18. 18.
    Jithendra P, Rajam AM, Kalaivani T, Mandal AB, Rose C. Preparation and characterization of Aloe vera blended collagen-chitosan composite scaffold for tissue engineering applications. ACS Appl Mater Interfaces. 2013;5:7291–8.CrossRefGoogle Scholar
  19. 19.
    Tummalapalli M, Berthet M, Verrier B, Deopura BL, Alam MS, Gupta B. Composite wound dressings of pectin and gelatin with Aloe vera and curcumin as bioactive agents. Int J Biol Macromol. 2016;82:104–13.CrossRefGoogle Scholar
  20. 20.
    Rodríguez E, Darias J, Díaz C. Aloe vera as a functional ingredient in foods. Crit Rev Food Sci Nutr. 2010;50:305–26.CrossRefGoogle Scholar
  21. 21.
    Moreno M, Isla MI, Sampietro AR, Vattuone MA. Comparison of the free radical-scavenging activity of propolis from several regions of Argentina. J Ethnopharmacol. 2000;71:109–14.CrossRefGoogle Scholar
  22. 22.
    Brilla E, Schosinsky K, Esquivel JM, Chavarria M. Cuantificacion de la glucosa por el metodo de la o-toluidina. Acta Med Cost. 1977;20:18–23.Google Scholar
  23. 23.
    USP (The United States Pharmacopeia). Water determination. In: Pharmacopeia—national formulary [USP 39 NF 34]. Rockville: United States Pharmacopeial Convention, Inc; 2013. p. 740.Google Scholar
  24. 24.
    USP (The United States Pharmacopeia). Residue on ignition. In: Pharmacopeia—national formulary [USP 39 NF 34]. Rockville: United States Pharmacopeial Convention, Inc; 2013. p. 308.Google Scholar
  25. 25.
    ISO. The International Organization for Standardization 10993-5: biological evaluation of medical devices. Part 5: test for in vitro cytotoxicity. 2009. p.1–24.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.
    Fontanilla MR, Casadiegos S, Bustos RH, Patarroyo MA. Comparison of healing of full-thickness skin wounds grafted with multidirectional or unidirectional autologous artificial dermis: differential delivery of healing biomarkers. Drug Deliv Transl Res. 2018;8:1014–24. Scholar
  28. 28.
    Bustos RH, Suesca E, Millan D, Gonzales 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;86:2421–8.CrossRefGoogle Scholar
  29. 29.
    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 A. 2012;18:1857–66.CrossRefGoogle Scholar
  30. 30.
    Barrantes E, Guinea M. Inhibition of collagenase and metalloproteinases by aloins and aloe gel. Life Sci. 2003;72:843–50.CrossRefGoogle Scholar
  31. 31.
    Starcher B. A ninhydrin-based assay to quantitate the total protein content of tissue samples. Anal Biochem. 2001;292:125–9.CrossRefGoogle Scholar
  32. 32.
    Council NR. Guide for the care and use of laboratory animals. Eight ed. Washington, DC: The National Academies Press; 2011.Google Scholar
  33. 33.
    Harris C, Bates-Jensen B, Parslow N, Raizman R, Singh M, Ketchen R. Bates-Jensen wound assessment tool: pictorial guide validation project. J Wound Ostomy Cont Nurs. 2010;37:253–9.CrossRefGoogle Scholar
  34. 34.
    Leary S, Underwood W, Lilly E, Anthony R, Cartner S, Corey D, et al. AVMA guidelines for the euthanasia of animals: 2013 Edition. 2013. p. 5–98.Google Scholar
  35. 35.
    Eshun K, He Q. Aloe Vera: a valuable ingredient for the food, pharmaceutical and cosmetic industries—a review. Crit Rev Food Sci Nutr. 2004;44:91–6.CrossRefGoogle Scholar
  36. 36.
    Park MK, Park JH, Kim NY, Shin YG, Choi YS, Lee JG, et al. Analysis of 13 phenolic compounds in Aloe species by high performance liquid chromatography. Phytochem Anal. 1998;9:186–91.CrossRefGoogle Scholar
  37. 37.
    du Plessis LH, Hamman JH. In vitro evaluation of the cytotoxic and apoptogenic properties of aloe whole leaf and gel materials. Drug Chem Toxicol. 2014;37:169–77.CrossRefGoogle Scholar
  38. 38.
    Vidal CMP, Leme AA, Aguiar TR, Phansalkar R, Nam J-W, Bisson J, et al. Mimicking the hierarchical functions of dentin collagen cross-links with plant derived phenols and phenolic acids. Langmuir. 2014;30:14887–93.CrossRefGoogle Scholar
  39. 39.
    Walter R, Miguez PA, Arnold RR, Pereira PNR, Duarte WR, Yamauchi M. Effects of natural cross-linkers on the stability of dentin collagen and the inhibition of root caries in vitro. Caries Res. 2008;42:263–8.CrossRefGoogle Scholar
  40. 40.
    Han B, Jaurequi J, Tang BW, Nimni ME. Proanthocyanidin: a natural crosslinking reagent for stabilizing collagen matrices. J Biomed Mater Res A. 2003;65A:118–24.CrossRefGoogle Scholar
  41. 41.
    McManus JP, Davis KG, Beart JE, Gaffney SH, Lilley TH, Haslam E. Polyphenol interactions. Part 1. Introduction; some observations on the reversible complexation of polyphenols with proteins and polysaccharides. J Chem Soc Perkin Trans. 1985;2:1429–38.CrossRefGoogle Scholar
  42. 42.
    Tanzer ML. Cross-linking of collagen. Science. 1973;180:561–6.CrossRefGoogle Scholar
  43. 43.
    Yates CC, Whaley D, Kulasekeran P, Hancock WW, Lu B, Bodnar R, et al. Delayed and deficient dermal maturation in mice lacking the CXCR3 ELR-negative CXC chemokine receptor. Am J Pathol. 2007;171:484–95.CrossRefGoogle Scholar
  44. 44.
    Gabbiani G. The myofibroblast in wound healing and fibrocontractive diseases. J Pathol. 2003;200:500–3.CrossRefGoogle Scholar
  45. 45.
    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;6:57–66.CrossRefGoogle Scholar
  46. 46.
    Wang C, Wang Q, Gao W, Zhang Z, Lou Y, Jin H, et al. Highly efficient local delivery of endothelial progenitor cells significantly potentiates angiogenesis and full-thickness wound healing. Acta Biomater. 2018;69:156–69.CrossRefGoogle Scholar
  47. 47.
    Jimi S, Kimura M, De Francesco F, Riccio M, Hara S, Ohjimi H. Acceleration mechanisms of skin wound healing by autologous micrograft in mice. Int J Mol Sci. 2017;18:1675.CrossRefGoogle Scholar
  48. 48.
    Lammers G, Verhaegen P, Ulrich M, Schalkwijk J, Middelkoop E, Weiland D, et al. An overview of methods for the in vivo evaluation of tissue-engineered skin constructs. Tissue Eng B. 2011;17:33–54.CrossRefGoogle Scholar
  49. 49.
    Hermann P. The direction of growth of human epidermis. Br J Dermatol. 1970;83:556–64.CrossRefGoogle Scholar
  50. 50.
    Ye Q, van Amerongen MJ, Sandham JA, Bank RA, van Luyn MJ, Harmsen MC. Site-specific tissue inhibitor of metalloproteinase-1 governs the matrix metalloproteinases-dependent degradation of crosslinked collagen scaffolds and is correlated with interleukin-10. J Tissue Eng Regen Med. 2011;5:264–74.CrossRefGoogle Scholar
  51. 51.
    Eming SA, Krieg T, Davidson JM. Inflammation in wound repair: molecular and cellular mechanisms. J Invest Dermatol. 2007;127:514–25.CrossRefGoogle Scholar
  52. 52.
    Janis JE, Harrison B. Wound healing. Part I. Basic science. Plast Reconstr Surg. 2014;133:199e–207e.CrossRefGoogle Scholar
  53. 53.
    Velnar T, Bailey T, Smrkolj V. The wound healing process: an overview of the cellular and molecular mechanisms. J Int Med Res. 2009;37:1528–42.CrossRefGoogle Scholar

Copyright information

© Controlled Release Society 2018

Authors and Affiliations

  1. 1.Tissue Engineering Group, Department of PharmacyUniversidad Nacional de ColombiaBogotáColombia

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