Publicação
Keratin peptides from chicken feathers for biomedical application
| Resumo: | There are several skin disorders, like eczema, psoriasis, acne, chronic wounds …and all of them involved several steps for skin regeneration. Depending on the skin disorder it is important to stimulate or delay keratinocytes migration and proliferation. It is known that keratin peptides have an influence of the metabolism of skin cells. Keratin can be obtained from several sources, like nails or hair. These sources have a limited supply, so it is important to obtain a new source for bioactive keratin peptides. Chicken feather are mainly composed by keratin and its disposal is a critical environmental problem. Through chicken feathers hydrolyze it is possible to obtain high value keratin peptides minimizing the impact on the environment and giving value to otherwise waste material. On this project, chicken feathers were hydrolyzed by keratinolytic enzymes produced by different bacteria strains (S196D, S195A, S59D, S188D, CD1 and S194B). After each fermentation the peptides were subjected to different purification procedures: stage1- the peptides were isolated using a C18 column; stage2 - eluted using a cut-off of 5kDA and fractioned with different percentages of acetonitrile 20% and 40% and finally stage3- each fraction was eluted using a Superdex peptide column. At each stage the of the purification 45 μg/mL of the peptides were placed in contact with skin cells (keratinocytes). The influence of the keratin peptides on cell migration and cell proliferation was evaluated over time. The anti-inflammatory activity of each peptide was evaluated using macrophages. The peptides were subjected to final purification prior to their contact with the cells; they were filtered. As this step could influence the composition of peptidic solution, the bioactivity of the non-filtered peptidic solution was also evaluated. The composition of the keratin peptides was analyzed by LC-MS/MS. The peptides subjected to the Stage 2 purification P188D 20%, P188D 40% (filtered and unfiltered), induced a decrease on keratinocytes migration. In Stage 3, all P188D 20 % peptides, enhanced cell migration, both filtered and unfiltered, with the exception of P188D 20% P2 that had no effect when filtered. The P188D 40% filtered solution promoted an increase on cell migration over 30%. These peptides do not seem to have anti-inflammatory activity. The chicken feathers keratin peptidic solution obtained and characterized on this project will have an important role on the treatment of different in skin disorders. |
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| Autores principais: | Dinis, Hugo Francisco Moreira |
| Assunto: | Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
| Ano: | 2018 |
| 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: | There are several skin disorders, like eczema, psoriasis, acne, chronic wounds …and all of them involved several steps for skin regeneration. Depending on the skin disorder it is important to stimulate or delay keratinocytes migration and proliferation. It is known that keratin peptides have an influence of the metabolism of skin cells. Keratin can be obtained from several sources, like nails or hair. These sources have a limited supply, so it is important to obtain a new source for bioactive keratin peptides. Chicken feather are mainly composed by keratin and its disposal is a critical environmental problem. Through chicken feathers hydrolyze it is possible to obtain high value keratin peptides minimizing the impact on the environment and giving value to otherwise waste material. On this project, chicken feathers were hydrolyzed by keratinolytic enzymes produced by different bacteria strains (S196D, S195A, S59D, S188D, CD1 and S194B). After each fermentation the peptides were subjected to different purification procedures: stage1- the peptides were isolated using a C18 column; stage2 - eluted using a cut-off of 5kDA and fractioned with different percentages of acetonitrile 20% and 40% and finally stage3- each fraction was eluted using a Superdex peptide column. At each stage the of the purification 45 μg/mL of the peptides were placed in contact with skin cells (keratinocytes). The influence of the keratin peptides on cell migration and cell proliferation was evaluated over time. The anti-inflammatory activity of each peptide was evaluated using macrophages. The peptides were subjected to final purification prior to their contact with the cells; they were filtered. As this step could influence the composition of peptidic solution, the bioactivity of the non-filtered peptidic solution was also evaluated. The composition of the keratin peptides was analyzed by LC-MS/MS. The peptides subjected to the Stage 2 purification P188D 20%, P188D 40% (filtered and unfiltered), induced a decrease on keratinocytes migration. In Stage 3, all P188D 20 % peptides, enhanced cell migration, both filtered and unfiltered, with the exception of P188D 20% P2 that had no effect when filtered. The P188D 40% filtered solution promoted an increase on cell migration over 30%. These peptides do not seem to have anti-inflammatory activity. The chicken feathers keratin peptidic solution obtained and characterized on this project will have an important role on the treatment of different in skin disorders. |
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