ORIGINAL PAPER Designing a gelatin/chitosan/hyaluronic acid biopolymer using a thermophysical approach for use in tissue engineering Javier Enrione • Paulo Dı ´az-Caldero ´n • Caroline R. Weinstein-Oppenheimer • Elizabeth Sa ´nchez • Miguel A. Fuentes • Donald I. Brown • Hugo Herrera • Cristian A. Acevedo Received: 24 April 2013 / Accepted: 7 May 2013 / Published online: 25 May 2013 Ó Springer-Verlag Berlin Heidelberg 2013 Abstract Cell culture on biopolymeric scaffolds has provided treatments for tissue engineering. Biopolymeric mixtures based on gelatin (Ge), chitosan (Ch) and hyalu- ronic acid (Ha) have been used to make scaffolds for wound healing. Thermal and physical properties of scaf- folds 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 experi- ment 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 char- acterization may be used to design scaffolds for tissue engineering. Keywords Biopolymeric scaffold Á Chitosan Á Gelatin Á Hyaluronic acid Á Thermophysical properties Á Tissue engineering Introduction Cell culture on biopolymeric scaffolds has provided a range of promising therapeutic treatments for tissue engi- neering allowing proliferation of several cell types such as fibroblasts [1], keratinocytes [2], chondrocytes [3] and stem cells [4, 5]. The use of scaffolds to seed cells using dif- ferent types of biomaterials, such as gelatin (Ge) or chitosan (Ch) recreates a 3D environment that mimics in part the geometry and chemistry of an extra-cellular matrix. The main effect produced by the use of scaffolds is that cells find a more suitable microenvironment compared to traditional monolayer culture methods. Traditional methods of cell growth in 2D have proven to be less effi- cient for the growth of cells with productive or therapeutic purposes. Biopolymer mixtures based on Ge/Ch have been used to make scaffolds for tissue engineering, alone or in combi- nation with other materials [1, 2, 5–11]. The addition of hyaluronic acid (Ha) in the preparation of Ge/Ch-scaffolds, firstly reported by Liu et al. [6], allowed obtaining an J. Enrione Á P. Dı ´az-Caldero ´n Faculty of Medicine, School of Nutrition and Dietetics, School of Service Management, Universidad de los Andes, San Carlos de Apoquindo 2200, Las Condes, Santiago, Chile P. Dı ´az-Caldero ´n Departamento de Ciencia y Tecnologı ´a de los Alimentos, Facultad Tecnolo ´gica, Universidad de Santiago de Chile (USACH), Avenida Ecuador 3769, Santiago, Chile C. R. Weinstein-Oppenheimer Departamento de Bioquı ´mica, Facultad de Farmacia, Universidad de Valparaı ´so, Avenida Gran Bretan ˜a 1093, Valparaı ´so, Chile E. Sa ´nchez Á M. A. Fuentes Á C. A. Acevedo (&) Centro de Biotecnologı ´a, Universidad Te ´cnica Federico Santa Marı ´a, Avenida Espan ˜a 1680, Valparaı ´so, Chile e-mail: cristian.acevedo@usm.cl D. I. Brown Á H. Herrera Departamento de Biologı ´a y Ciencias Ambientales, Facultad de Ciencias, Universidad de Valparaı ´so, Avenida Gran Bretan ˜a 1111, Valparaı ´so, Chile 123 Bioprocess Biosyst Eng (2013) 36:1947–1956 DOI 10.1007/s00449-013-0971-x