Standardization of organoid culture for evaluation of melanogenesis induced by UVB, UVA and visible light,
Abstract Background: Organoid cultures are primary cultures that maintain architectural characteristics and the relationships between cells, as well as the extracellular matrix. They are alternatives for pathophysiological or therapeutic investigation rather than animal and in vitro tests. Objecti...
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| Format: | Article |
| Language: | English |
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Sociedade Brasileira de Dermatologia
2020-03-01
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| Series: | Anais Brasileiros de Dermatologia |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0365-05962020000100046&tlng=en |
| _version_ | 1818987749832654848 |
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| author | Thainá Oliveira Felicio Olivatti Giovana Piteri Alcantara Ana Cláudia Cavalcante Espósito Lemos Márcia Guimarães da Silva Hélio Amante Miot |
| author_facet | Thainá Oliveira Felicio Olivatti Giovana Piteri Alcantara Ana Cláudia Cavalcante Espósito Lemos Márcia Guimarães da Silva Hélio Amante Miot |
| author_sort | Thainá Oliveira Felicio Olivatti |
| collection | DOAJ |
| description | Abstract Background: Organoid cultures are primary cultures that maintain architectural characteristics and the relationships between cells, as well as the extracellular matrix. They are alternatives for pathophysiological or therapeutic investigation rather than animal and in vitro tests. Objective: Development of a cutaneous organoid culture model, aiming at the study of radiation-induced melanogenesis. Method: A validation study, which involved biopsies of the skin of the back of the adult ear. One sample was irradiated with different doses of UVB, UVA, or visible light (VL); the other was maintained in the dark for 72 h. The viability of the tissues was evaluated from the morphological and architectural parameters of the histology, and the expression of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene, by real-time polymerase chain reaction (PCR). The radiation-induced melanin pigmentation was standardized according to the doses of each radiation and evaluated by digital image analysis (Fontana-Masson). Results: The primary skin culture was standardized at room temperature using DMEM medium. The doses of UVB, UVA, and VL (blue light) that induced differential melanogenesis were: 166 mJ/cm2, 1.524 J/cm2, and 40 J/cm2. The expression of the GAPHD constitutional gene did not differ between the sample of skin processed immediately after tissue collection and the sample cultured for 72 h in the standardized protocol. Study limitations: This was a preliminary study that evaluated only the viability and integrity of the melanogenic system, and the effect of the radiation alone. Conclusions: The standardized model maintained viable melanocytic function for 72 h at room temperature, allowing the investigation of melanogenesis induced by different forms of radiation. |
| first_indexed | 2024-12-20T19:11:38Z |
| format | Article |
| id | doaj.art-7983d6d2f3da4e139f57489ea70ba397 |
| institution | Directory Open Access Journal |
| issn | 0365-0596 |
| language | English |
| last_indexed | 2024-12-20T19:11:38Z |
| publishDate | 2020-03-01 |
| publisher | Sociedade Brasileira de Dermatologia |
| record_format | Article |
| series | Anais Brasileiros de Dermatologia |
| spelling | doaj.art-7983d6d2f3da4e139f57489ea70ba3972022-12-21T19:29:12ZengSociedade Brasileira de DermatologiaAnais Brasileiros de Dermatologia0365-05962020-03-01951465110.1016/j.abd.2019.06.005Standardization of organoid culture for evaluation of melanogenesis induced by UVB, UVA and visible light,Thainá Oliveira Felicio Olivattihttps://orcid.org/0000-0002-4719-4821Giovana Piteri Alcantarahttps://orcid.org/0000-0001-6749-7160Ana Cláudia Cavalcante Espósito Lemoshttps://orcid.org/0000-0001-9283-2354Márcia Guimarães da Silvahttps://orcid.org/0000-0002-4663-7926Hélio Amante Miothttps://orcid.org/0000-0002-2596-9294Abstract Background: Organoid cultures are primary cultures that maintain architectural characteristics and the relationships between cells, as well as the extracellular matrix. They are alternatives for pathophysiological or therapeutic investigation rather than animal and in vitro tests. Objective: Development of a cutaneous organoid culture model, aiming at the study of radiation-induced melanogenesis. Method: A validation study, which involved biopsies of the skin of the back of the adult ear. One sample was irradiated with different doses of UVB, UVA, or visible light (VL); the other was maintained in the dark for 72 h. The viability of the tissues was evaluated from the morphological and architectural parameters of the histology, and the expression of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene, by real-time polymerase chain reaction (PCR). The radiation-induced melanin pigmentation was standardized according to the doses of each radiation and evaluated by digital image analysis (Fontana-Masson). Results: The primary skin culture was standardized at room temperature using DMEM medium. The doses of UVB, UVA, and VL (blue light) that induced differential melanogenesis were: 166 mJ/cm2, 1.524 J/cm2, and 40 J/cm2. The expression of the GAPHD constitutional gene did not differ between the sample of skin processed immediately after tissue collection and the sample cultured for 72 h in the standardized protocol. Study limitations: This was a preliminary study that evaluated only the viability and integrity of the melanogenic system, and the effect of the radiation alone. Conclusions: The standardized model maintained viable melanocytic function for 72 h at room temperature, allowing the investigation of melanogenesis induced by different forms of radiation.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0365-05962020000100046&tlng=enMelanosisOrganoidsPhotobiology |
| spellingShingle | Thainá Oliveira Felicio Olivatti Giovana Piteri Alcantara Ana Cláudia Cavalcante Espósito Lemos Márcia Guimarães da Silva Hélio Amante Miot Standardization of organoid culture for evaluation of melanogenesis induced by UVB, UVA and visible light, Anais Brasileiros de Dermatologia Melanosis Organoids Photobiology |
| title | Standardization of organoid culture for evaluation of melanogenesis induced by UVB, UVA and visible light, |
| title_full | Standardization of organoid culture for evaluation of melanogenesis induced by UVB, UVA and visible light, |
| title_fullStr | Standardization of organoid culture for evaluation of melanogenesis induced by UVB, UVA and visible light, |
| title_full_unstemmed | Standardization of organoid culture for evaluation of melanogenesis induced by UVB, UVA and visible light, |
| title_short | Standardization of organoid culture for evaluation of melanogenesis induced by UVB, UVA and visible light, |
| title_sort | standardization of organoid culture for evaluation of melanogenesis induced by uvb uva and visible light |
| topic | Melanosis Organoids Photobiology |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0365-05962020000100046&tlng=en |
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