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Collection and culture of human connective tissue cells from gingival explant technique for oral tissue bioengineering / Colección de células de tejido conectivo humano a partir de la técnica de explante gingival para bioingeniería de tejidos orales
Abreu, Fernando Antônio Mauad de; Reis, Igor Daniel Garcia; Silva, Gerluza Aparecida Borges; Jorge, Erika Cristina.
  • Abreu, Fernando Antônio Mauad de; Federal University of Minas Gerais. Biological Sciences Institute. Department of Morphology. Belo Horizonte. BR
  • Reis, Igor Daniel Garcia; Federal University of Minas Gerais. Biological Sciences Institute. Department of Morphology. Belo Horizonte. BR
  • Silva, Gerluza Aparecida Borges; Federal University of Minas Gerais. Biological Sciences Institute. Department of Morphology. Belo Horizonte. BR
  • Jorge, Erika Cristina; Federal University of Minas Gerais. Biological Sciences Institute. Department of Morphology. Belo Horizonte. BR
Int. j. morphol ; 37(4): 1229-1233, Dec. 2019. graf
Article in English | LILACS | ID: biblio-1040117
Responsible library: CL1.1
ABSTRACT

SUMMARY:

Cell culture is an important tool in medical, odontological and biological research laboratories, supporting cell therapies and tissue bioengineering strategies. Gingival fibroblasts present structural function, being able to modulate their metabolic capacity, which is reflected in the tissue morphology. The possibility of culturing fibroblasts in vitro, in monolayer or on three-dimensional scaffolds, for subsequent transplants in vivo opens important perspectives for the periodontal surgical clinic. The objective of the present article is to present a method of obtaining and cultivating viable human gingival fibroblasts for in vitro research. Explants derived from periodontal surgical discards were used, grown in 25 cm2 bottles to obtain a primary cell culture. After observing the proliferation and growth of the fibroblasts that interconnected and formed a monolayer network, involving the periphery of the explants, it was possible to remove the explants, to make the passage and the new subcultures were obtained in a ratio of 11. After 7 days, the amount of viable cells was analyzed in triplicate, using the Neubauer chamber technique, in cell culture bottles of 25 mm2 (T25) and 75 mm2 (T75). Fibroblasts were described and subclassified morphologically. The results showed a growth pattern in both bottles, but with a larger number in bottles of 75 cm2. Cells with fibroblastic morphology were subclassified into reticular and fusiform, being predominant those with fusiform morphology. In conclusion, culture of explant of human gingival connective tissue is a viable method for obtaining gingival connective tissue cells suitable for laboratory tests in cell culture, aiming at obtaining constructs for gingival tissue engineering.
RESUMEN
RESUMEN El cultivo celular es una herramienta importante en los laboratorios de investigación médica, odontológica y biológica, que apoyan las terapias celulares y las estrategias de bioingeniería de tejidos. Los fibroblastos gingivales presentan una función estructural, pudiendo modular su capacidad metabólica, que se refleja en la morfología tisular. La posibilidad de cultivar fibroblastos in vitro, en monocapa o en andamios tridimensionales, para trasplantes posteriores in vivo abre perspectivas importantes para la clínica de cirugía periodontal. El objetivo del presente artículo es presentar un método para obtener y cultivar fibroblastos gingivales humanos viables para investigación in vitro. Se utilizaron explantes derivados de los descartes quirúrgicos periodontales, crecidos en frascos de 25 cm2 para obtener un cultivo de células primarias. Después de observar la proliferación y el crecimiento de los fibroblastos que se interconectaron y formaron una red de monocapa, que involucraba la periferia de los explantes, fue posible eliminar los explantes, hacer el pasaje y los nuevos subcultivos se obtuvieron en una proporción de 11. Después de 7 días, la cantidad de células viables se analizó por triplicado, utilizando la técnica de cámara de Neubauer, en botellas de cultivo celular de 25 mm2 (T25) y 75 mm2 (T75). Los fibroblastos fueron descritos y sub-clasificados morfológicamente. Los resultados mostraron un patrón de crecimiento en ambas botellas, pero con un número mayor en botellas de 75 cm2. Las células con morfología fibroblástica se subclasificaron en reticulares y fusiformes, predominando aquellas con morfología fusiforme. En conclusión, el cultivo de explante de tejido conectivo gingival humano es un método viable para obtener células de tejido conectivo gingival adecuadas para pruebas de laboratorio en cultivos celulares, con el objetivo de obtener construcciones para la ingeniería del tejido gingival.
Subject(s)

Full text: Available Index: LILACS (Americas) Main subject: Connective Tissue Cells / Cell Culture Techniques / Bioengineering / Gingiva Limits: Humans Language: English Journal: Int. j. morphol Journal subject: Anatomy Year: 2019 Type: Article Affiliation country: Brazil Institution/Affiliation country: Federal University of Minas Gerais/BR

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Full text: Available Index: LILACS (Americas) Main subject: Connective Tissue Cells / Cell Culture Techniques / Bioengineering / Gingiva Limits: Humans Language: English Journal: Int. j. morphol Journal subject: Anatomy Year: 2019 Type: Article Affiliation country: Brazil Institution/Affiliation country: Federal University of Minas Gerais/BR