Mamatha Yadlapati1, Claudia Biguetti2, Franco Cavalla3, Francisco Nieves1, Christopher Bessey1, Pedram Bohluli1, Gustavo Pompermaier Garlet4, Ariadne Letra5, Walid D Fakhouri6, Renato Menezes Silva7. 1. Department of Endodontics, University of Texas School of Dentistry at Houston, Houston, Texas. 2. Department of Biological Sciences, School of Dentistry of Bauru, University of São Paulo (FOB/USP), Bauru, São Paulo, Brazil; Center for Craniofacial Research, University of Texas School of Dentistry at Houston, Houston, Texas. 3. Department of Biological Sciences, School of Dentistry of Bauru, University of São Paulo (FOB/USP), Bauru, São Paulo, Brazil; Center for Craniofacial Research, University of Texas School of Dentistry at Houston, Houston, Texas; Department of Conservative Dentistry, University of Chile School of Dentistry, Santiago, Chile. 4. Department of Biological Sciences, School of Dentistry of Bauru, University of São Paulo (FOB/USP), Bauru, São Paulo, Brazil. 5. Department of Endodontics, University of Texas School of Dentistry at Houston, Houston, Texas; Center for Craniofacial Research, University of Texas School of Dentistry at Houston, Houston, Texas. 6. Center for Craniofacial Research, University of Texas School of Dentistry at Houston, Houston, Texas; Department of Diagnostic and Biomedical Sciences, University of Texas School of Dentistry at Houston, Houston, Texas. 7. Department of Endodontics, University of Texas School of Dentistry at Houston, Houston, Texas; Center for Craniofacial Research, University of Texas School of Dentistry at Houston, Houston, Texas. Electronic address: Renato.M.Silva@uth.tmc.edu.
Abstract
INTRODUCTION: Vascular endothelial growth factor (VEGF) is a signal protein that stimulates angiogenesis and vasculogenesis and has been used in tissue regeneration and pulp regeneration experimental models. The purpose of this study was to develop a delivery system composed of a biodegradable fiber and controlled release of VEGF to promote cell viability and secure an adequate blood supply for the survival of human stem cells of the apical papilla (SCAP) favoring endodontic regenerative procedures. METHODS: We developed a polydioxanone fiber, 50 μm in diameter, loaded with VEGF at a linear concentration of 12.2 ng/cm. Cytotoxic effects of the VEGF-loaded fiber (VF) on SCAP and mouse fibroblasts were assessed by using a multiparametric assay kit (XTT-NR-CVDE [Xenometrix, Allschwil, Switzerland]). We evaluated VF-induced mRNA expression of downstream growth factors by using a human growth factor Taqman array in real-time polymerase chain reaction. We also assessed the in vivo subcutaneous reaction of C57BL/6 mice to implants of VF alone and human root fragments (10 mm in length) filled with VF after 10, 20, and 45 days. Statistical analyses were performed by using analysis of variance and Student t tests or non-parametric alternatives. RESULTS: Enzyme-linked immunosorbent assay verified detectable concentrations of released VEGF in solution for 25 days. No cytotoxicity was observed on SCAP and mouse fibroblasts treated with VEGF. In addition, VEGF treatment also induced the expression of additional growth factors with roles in tissue and blood vessel formation and neuroprotective function. Implantation of VF and root fragments filled with VF showed biocompatibility in vivo, promoting new blood vessels and connective tissue formation into the root canal space with negligible inflammation. CONCLUSIONS: Our results show that the VF used in this study is biocompatible and may be a promising scaffold for additional optimization and use in endodontic regenerative procedures.
INTRODUCTION:Vascular endothelial growth factor (VEGF) is a signal protein that stimulates angiogenesis and vasculogenesis and has been used in tissue regeneration and pulp regeneration experimental models. The purpose of this study was to develop a delivery system composed of a biodegradable fiber and controlled release of VEGF to promote cell viability and secure an adequate blood supply for the survival of human stem cells of the apical papilla (SCAP) favoring endodontic regenerative procedures. METHODS: We developed a polydioxanone fiber, 50 μm in diameter, loaded with VEGF at a linear concentration of 12.2 ng/cm. Cytotoxic effects of the VEGF-loaded fiber (VF) on SCAP and mouse fibroblasts were assessed by using a multiparametric assay kit (XTT-NR-CVDE [Xenometrix, Allschwil, Switzerland]). We evaluated VF-induced mRNA expression of downstream growth factors by using a human growth factor Taqman array in real-time polymerase chain reaction. We also assessed the in vivo subcutaneous reaction of C57BL/6 mice to implants of VF alone and human root fragments (10 mm in length) filled with VF after 10, 20, and 45 days. Statistical analyses were performed by using analysis of variance and Student t tests or non-parametric alternatives. RESULTS: Enzyme-linked immunosorbent assay verified detectable concentrations of released VEGF in solution for 25 days. No cytotoxicity was observed on SCAP and mouse fibroblasts treated with VEGF. In addition, VEGF treatment also induced the expression of additional growth factors with roles in tissue and blood vessel formation and neuroprotective function. Implantation of VF and root fragments filled with VF showed biocompatibility in vivo, promoting new blood vessels and connective tissue formation into the root canal space with negligible inflammation. CONCLUSIONS: Our results show that the VF used in this study is biocompatible and may be a promising scaffold for additional optimization and use in endodontic regenerative procedures.
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Authors: Sueli Patricia Harumi Miyagi de Cara; Clarice Silvia Taemi Origassa; Fernando de Sá Silva; Maria Stella N A Moreira; Danilo Candido de Almeida; Ana Clara Fagundes Pedroni; Giovanna Lopes Carvalho; Diego Pulzatto Cury; Niels Olsen Saraiva Câmara; Márcia Martins Marques Journal: Heliyon Date: 2019-04-28