Publicação
Development of hyaluronic acid, dextrin and extracellular matrix hydrogels for cell expansion
| Resumo: | Every day thousands of surgical procedures are performed to replace or repair tissue that has been damaged through disease or trauma. The developing field of Tissue Engineering (TE) aims to regenerate damaged tissues by combining cells from the body with highly porous scaffold biomaterials, which act as templates for tissue regeneration, to guide the growth of new tissue. Cells, scaffolds and growth-stimulating signals are generally referred to as the TE triad, the key components of engineered tissues. Scaffolds, typically made of polymeric biomaterials, provide the structural support for cell attachment and subsequent tissue development. However, researchers often encounter an enormous variety of choices when selecting scaffolds for Tissue Engineering. The aim of this project is to develop a new hydrogel made of Dextrin (Dex), Hyaluronic acid (HA) and Extracellular matrix (ECM) from Small Intestine Submucosa (SIS), using different types of HA, mixed different proportions. In this study, HA and Dex were oxidized by sodium periodate to create aldehyde functional groups, which could be cross-linked by Adipic Acid Dihidrazide (ADH). Their characterization was performed based on gelation period and degradation rate. In addition, cell viability tests were performed through a Resazurin assay, a MTS assay and Live and Dead (LD) using osteoblastic cell line MC3T3 calvaria from mouse. Results showed a low gelation time for all the hydrogels and low degradation rates for mixed hydrogels with high contents of high molecular weight (MW) HA. The degradation tests demonstrated that the selected hydrogel could maintain the gel matrix over 70 days. Interactions between the hydrogel, MC3T3 cells, and the extracellular matrix (ECM) were also evaluated, as were the effects of the hydrogels on MC3T3 cell growth. The hydrogels possess several clinical advantages, including sterilizability and rapid gelation time for injection. Cultured MC3T3 cells did not adhere to hydrogels, but survived and maintained their round morphology, evidencing that the hydrogel is biocompatible, but does not promote cell growth. |
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| Autores principais: | Carvalho, Alberto Dimas Fernandes Leite de |
| Assunto: | Engenharia e Tecnologia::Biotecnologia Industrial |
| Ano: | 2015 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade do Minho |
| Idioma: | inglês |
| Origem: | RepositóriUM - Universidade do Minho |
| Resumo: | Every day thousands of surgical procedures are performed to replace or repair tissue that has been damaged through disease or trauma. The developing field of Tissue Engineering (TE) aims to regenerate damaged tissues by combining cells from the body with highly porous scaffold biomaterials, which act as templates for tissue regeneration, to guide the growth of new tissue. Cells, scaffolds and growth-stimulating signals are generally referred to as the TE triad, the key components of engineered tissues. Scaffolds, typically made of polymeric biomaterials, provide the structural support for cell attachment and subsequent tissue development. However, researchers often encounter an enormous variety of choices when selecting scaffolds for Tissue Engineering. The aim of this project is to develop a new hydrogel made of Dextrin (Dex), Hyaluronic acid (HA) and Extracellular matrix (ECM) from Small Intestine Submucosa (SIS), using different types of HA, mixed different proportions. In this study, HA and Dex were oxidized by sodium periodate to create aldehyde functional groups, which could be cross-linked by Adipic Acid Dihidrazide (ADH). Their characterization was performed based on gelation period and degradation rate. In addition, cell viability tests were performed through a Resazurin assay, a MTS assay and Live and Dead (LD) using osteoblastic cell line MC3T3 calvaria from mouse. Results showed a low gelation time for all the hydrogels and low degradation rates for mixed hydrogels with high contents of high molecular weight (MW) HA. The degradation tests demonstrated that the selected hydrogel could maintain the gel matrix over 70 days. Interactions between the hydrogel, MC3T3 cells, and the extracellular matrix (ECM) were also evaluated, as were the effects of the hydrogels on MC3T3 cell growth. The hydrogels possess several clinical advantages, including sterilizability and rapid gelation time for injection. Cultured MC3T3 cells did not adhere to hydrogels, but survived and maintained their round morphology, evidencing that the hydrogel is biocompatible, but does not promote cell growth. |
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