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Development and characterization of a new chitosan-based scaffold associated with gelatin,microparticulate dentin and genipin for endodontic regeneration
Authors:Hector Caballero-Flores  Cleber Keiti Nabeshima  Giovanna Sarra  Maria Stella Moreira  Victor Elias Arana-Chavez  Márcia Martins Marques  Manoel Eduardo de Lima Machado
Affiliation:1. Discipline of Endodontic, Department of Restorative Dentistry, School of Dentistry, University of São Paulo, São Paulo, SP, Brazil;2. Post Graduation Program, School of Dentistry, Ibirapuera University, São Paulo, SP, Brazil;3. Department of Biomaterials and Oral Biology, School of Dentistry, University of São Paulo. São Paulo, SP, Brazil
Abstract:ObjectiveAn ideal scaffold for endodontic regeneration should allow the predictableness of the new tissue organization and limit the negative impact of residual bacteria. Therefore, composition and functionalization of the scaffold play an important role in tissue bioengineering. The objective of this study was to assess the morphological, physicochemical, biological and antimicrobial properties of a new solid chitosan-based scaffold associated with gelatin, microparticulate dentin and genipin.MethodsScaffolds based on chitosan (Ch); chitosan associated with gelatin and genipin (ChGG); and chitosan associated with gelatin, microparticulate dentin and genipin (ChGDG) were prepared by using the freeze-drying method. The morphology of the scaffolds was analyzed by scanning electron microscopy (SEM). The physicochemical properties were assessed for biodegradation, swelling and total released proteins. The biological aspects of the scaffolds were assessed using human cells from the apical papilla (hCAPs). Cell morphology and adhesion to the scaffolds were evaluated by SEM, cytotoxicity and cell proliferation by MTT reduction-assay. Cell differentiation in scaffolds was assessed by using alizarin red assay. The antimicrobial effect of the scaffolds was evaluated by using the bacterial culture method, and bacterial adhesion to the scaffolds was observed by SEM.ResultsAll the scaffolds presented porous structures. The ChCDG had more protein release, adhesion, proliferation and differentiation of hCAPs, and bacteriostatic effect on Enterococcus faecalis than Ch and ChGG (p < 0.05).SignificanceThe chitosan associated with gelatin, microparticulate dentin and genipin has morphological, physicochemical, biological and antibacterial characteristics suitable for their potential use as scaffold in regenerative endodontics.
Keywords:Tissue engineering  Cell differentiation  Dentin  Polymers  Chitosan  Genepin
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