Abstract
Cell culture on biopolymeric scaffolds has provided treatments for tissue engineering. Biopolymeric mixtures based on gelatin (Ge), chitosan (Ch) and hyaluronic acid (Ha) have been used to make scaffolds for wound healing. Thermal and physical properties of scaffolds prepared with Ge, Ch and Ha were characterized. Thermal characterization was made by using differential scanning calorimetry (DSC), and physical characterization by gas pycnometry and scanning electron microscopy. The effects of Ge content and cross-linking on thermophysical properties were evaluated by means of a factorial experiment design (central composite face centered). Gelatin content was the main factor that affects the thermophysical properties (microstructure and thermal transitions) of the scaffold. The effect of Ge content of the scaffolds for tissue engineering was studied by seeding skin cells on the biopolymers. The cell attachment was not significantly modified at different Ge contents; however, the cell growth rate increased linearly with the decrease of the Ge content. This relationship together with the thermophysical characterization may be used to design scaffolds for tissue engineering.
Similar content being viewed by others
References
Acevedo C, Somoza R, Weinstein-Oppenheimer C, Silva S, Moreno M, Sanchez E, Albornoz F, Young M, MacNaughtan W, Enrione J (2013) Improvement of human skin cell growth by radiation-induced modifications of a Ge/Ch/Ha scaffold. Bioprocess Biosyst Eng 36:317–324
Weinstein-Oppenheimer CR, Aceituno AR, Brown DI, Acevedo C, Ceriani R, Fuentes MA, Albornoz F, Henríquez-Roldán CF, Morales P, Maclean C, Tapia SM, Young ME (2010) The effect of an autologous cellular gel-matrix integrated implant system on wound healing. J Transl Med 8:59
Ko CL, Tien YC, Wang JC, Chen WC (2012) Characterization of controlled highly porous hyaluronan/gelatin cross-linking sponges for tissue engineering. J Mech Behav Biomed Mater 14:227–238
Li Z, Leung M, Hopper R, Ellenbogen R, Zhang M (2010) Feeder-free self-renewal of human embryonic stem cells in 3D porous natural polymer scaffolds. Biomaterials 31(3):404–412
Miranda SC, Silva GA, Hell RC, Martins MD, Alves JB, Goes AM (2011) Three-dimensional culture of rat BMMSCs in a porous chitosan-gelatin scaffold: a promising association for bone tissue engineering in oral reconstruction. Arch Oral Biol 56:1–15
Liu H, Mao J, Yao K, Yang G, Cui L, Cao Y (2004) A study on a chitosan-gelatin-hyaluronic acid scaffold as artificial skin in vitro and its tissue engineering applications. J Biomater Sci Polym Ed 15:25–40
Kathuria N, Tripathi A, Kar KK, Kumar A (2009) Synthesis and characterization of elastic and macroporous chitosan-gelatin cryogels for tissue engineering. Acta Biomater 5:406–418
Thein-Han WW, Saikhun J, Pholpramoo C, Misra RD, Kitiyanant Y (2009) Chitosan-gelatin scaffolds for tissue engineering: physico-chemical properties and biological response of buffalo embryonic stem cells and transfectant of GFP-buffalo embryonic stem cells. Acta Biomater 5:3453–3466
Enrione J, Osorio F, Lopez D, Weinstein-Oppenheimer C, Fuentes MA, Ceriani R, Brown DI, Albornoz F, Sanchez E, Villalobos P, Somoza RA, Young ME, Acevedo CA (2010) Characterization of a gelatin/chitosan/hyaluronan scaffold-polymer. Electron J Biotechnol 13:15
Iyer P, Walker KJ, Madihally SV (2012) Increased matrix synthesis by fibroblasts with decreased proliferation on synthetic chitosan-gelatin porous structures. Biotechnol Bioeng 109:1314–1325
Miranda SC, Silva GA, Mendes RM, Abreu FA, Caliari MV, Alves JB, Goes AM (2012) Mesenchymal stem cells associated with porous chitosan-gelatin scaffold: a potential strategy for alveolar bone regeneration. J Biomed Mater Res A 100:2775–2786
Chang CH, Liu HC, Lin CC, Chou CH, Lin FH (2003) Gelatin-chondroitin-hyaluronan tri-copolymer scaffold for cartilage tissue engineering. Biomaterials 24:4853–4858
Huang Y, Onyeri S, Siewe M, Moshfeghian A, Madihally SV (2005) In Vitro characterization of chitosan-gelatin scaffolds for tissue engineering. Biomaterials 26:7616–7627
Mao JS, Zhao LG, Yin YJ, Yao KD (2003) Structure and properties of bilayer chitosan-gelatin scaffolds. Biomaterials 24:1067–1074
Xia WY, Liu W, Cui L, Liu YC, Zhong W, Liu D, Wu J, Chua K, Cao Y (2004) Tissue engineering of cartilage with the use of chitosan-gelatin complex scaffolds. J Biomed Mater Res B Appl Biomater 71B:373–380
Lazaridou A, Biliaderis CG (2002) Thermophysical properties of chitosan, chitosan-starch and chitosan-pullulan films near the glass transition. Carbohydr Polym 48:179–190
Enrione JI, Sáez C, López D, Skurtys O, Acevedo C, Osorio F, MacNaughtan W, Hill S (2012) Structural relaxation of salmon gelatin films in the glassy state. Food Bioprocess Technol 5:2446–2453
Díaz P, López D, Matiacevich S, Osorio F, Enrione J (2011) State diagram of salmon (Salmo salar) gelatin films. J Sci Food Agric 91:2558–2565
Fox TG (1956) Influence of diluent and copolymer composition on the glass temperature of a polymer system. Bull Am Phys Soc 1:123
Couchman PR, Karasz FE (1978) A Classical Thermodynamic Discussion of the Effect of Composition on Glass-Transition Temperatures. Macromolecules 11:117–119
Heath DE, Cooper SL (2010) Interaction of endothelial cells with methacrylic terpolymer biomaterials. J Biomed Mater Res B Appl Biomater 92(2):289–297
Heath DE, Kobe C, Jones D, Moldovan NI, Cooper SL (2013) In vitro endothelialization of electrospun terpolymer scaffolds: evaluation of scaffold type and cell source. Tissue Eng Part A 19:79–90
Acevedo CA, Brown DI, Young ME, Reyes JG (2009) Senescent cultures of human dermal fibroblasts modified phenotype when immobilized in fibrin polymer. J Biomater Sci Polym Ed 20:1929–1942
Marreco PR, da Luz Moreira P, Genari SC, Moraes AM (2004) Effects of different sterilization methods on the morphology, mechanical properties, and cytotoxicity of chitosan membranes used as wound dressings. J Biomed Mater Res B Appl Biomater 71:268–277
Xiao J, Zhang Y, Wang J, Yu W, Wang W, Ma X (2010) Monitoring of cell viability and proliferation in hydrogel-encapsulated system by resazurin assay. Appl Biochem Biotechnol 162:1996–2007
Rahman MS, Al-Saidi G, Guizani N, Abdullah (2010) A Development of state diagram of bovine gelatin by measuring thermal characteristics using differential scanning calorimetry (DSC) and cooling curve method. Thermochim Acta 509:111–119
Yakimets I, Wellner N, Smith AC, Wilson RH, Farhat I, Mitchell J (2005) Mechanical properties with respect to water content of gelatin films in glassy state. Polymer 46:12577–12585
Sobral PJA, Menegalli FC, Hubinger MD, Roques MA (2001) Mechanical, water vapor barrier and thermal properties of gelatin based edible films. Food Hydrocoll 15:423–432
Badii F, Martinet C, Mitchell JR, Farhat I (2006) Enthalpy and mechanical relaxation of glassy gelatin films. Food Hydrocoll 20:879–884
Badii F, MacNaughtan W, Farhat IA (2005) Enthalpy relaxation of gelatin in the glassy state. Int J Biol Macromol 36:263–269
Coppola M, Djabourov M, Ferrand M (2008) Phase diagram of gelatin plasticized by water and glycerol. Macromol Symp 273:56–65
Slade L, Levine H (1995) Glass transition and water-food structure interactions. Adv Food Res 38:103–269
Lee YH, Lee JH, An IG, Kim C, Lee DS, Lee YK, Nam JD (2005) Electrospun dual-porosity structure and biodegradation morphology of Montmorillonite reinforced PLLA nanocomposite scaffolds. Biomaterials 26:3165–3172
Cooper JA, Lu HH, Ko FK, Freeman JW, Laurencin CT (2005) Fiber-based tissue-engineered scaffold for ligament replacement: design considerations and in vitro evaluation. Biomaterials 26:1523–1532
Huang Y, Onyeri S, Siewe M, Moshfeghian A, Madihally SV (2005) In vitro characterization of chitosan–gelatin scaffolds for tissue engineering. Biomaterials 26:7616–7627
Madihally SV, Matthew HW (1999) Porous chitosan scaffolds for tissue engineering. Biomaterials 20:1133–1142
Huang Y, Siewe M, Madihally SV (2006) Effect of spatial architecture on cellular colonization. Biotechnol Bioeng 93:64–75
Zhang F, He C, Cao L, Feng W, Wang H, Mo X, Wang J (2011) Fabrication of gelatin-hyaluronic acid hybrid scaffolds with tunable porous structures for soft tissue engineering. Int J Biol Macromol 48:474–481
Bowers KT, Keller JC, Randolph BA, Wick DG, Michaels CM (1992) Optimization of surface micromorphology for enhanced osteoblast responses in vitro. Int J Oral Maxillofac Implants 7:302–310
Anselme K (2000) Osteoblast adhesion on biomaterials. Biomaterials 21:667–681
Acknowledgments
The authors wish to thank CONICYT from Chile for FONDECYT Grant 1120166.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Enrione, J., Díaz-Calderón, P., Weinstein-Oppenheimer, C.R. et al. Designing a gelatin/chitosan/hyaluronic acid biopolymer using a thermophysical approach for use in tissue engineering. Bioprocess Biosyst Eng 36, 1947–1956 (2013). https://doi.org/10.1007/s00449-013-0971-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-013-0971-x