Cultura de condrócitos em arcabouço tridimensional: hidrogel de alginato
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Data
2009-01-01
Autores
Bittencourt, Renata Aparecida de Camargo [UNESP]
Pereira, Hamilton da Rosa [UNESP]
Felisbino, Sérgio Luís [UNESP]
Ferreira, Rosana Rossi [UNESP]
Guilherme, Gabrielle Reinoldes Bizarria [UNESP]
Moroz, Andrei[UNESP]
Deffune, Elenice [UNESP]
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Título de Volume
Editor
Sociedade Brasileira de Ortopedia e Traumatologia
Resumo
OBJETIVOS: O presente estudo teve como objetivo cultivar condrócitos retirados da articulação do joelho de coelhos encapsulados em hidrogel de alginato (HA) e caracterizar a produção de matriz extracelular (ECM). MÉTODOS: A cartilagem articular foi removida do joelho de coelhos, com três a seis meses, fragmentada em pedaços de 1mm e submetida à digestão enzimática. Uma concentração de 1x106 céls/mL foram ressuspensas em uma solução de alginato de sódio a 1,5% (w/v), em seguida fez-se o processo de gelatinização em CaCl2 (102 mM), permitindo a formação do HA e cultivo em meio DMEM-F12 durante quatro semanas. A distribuição das células e a ECM foram acessadas através das secções histológicas coradas com e azul de toluidina hematoxilina e eosina (HE). RESULTADOS: Houve um aumento no número e na viabilidade dos condrócitos durante as quatro semanas de cultura. Através das análises histológicas dos HAs corados com azul de toluidina e HE foi possível observar a distribuição definida dos condrócitos no hidrogel, assemelhando-se a grupos isógenos e formação de matriz territorial. CONCLUSÃO: Este estudo demonstrou a eficiência do HA como arcabouço para ser usado na cultura de condrócitos, constituindo uma alternativa no reparo de lesões na cartilagem articular.
OBJECTIVES: The aim of this study was to culture chondrocytes from knee joint cartilage of rabbits encapsulated in alginate hydrogel (HA) and to characterize the production of extracelular matrix (ECM). METHODS: Joint cartilage was obtained from rabbits' knees, three to six months old, fragmented into 1-mm pieces and submitted to enzymatic digestion. A concentration of 1x106 cells/mL were re-suspended into a 1.5% (w/v) sodium alginate solution, followed by gel formation process with CaCl2 (102 mM), allowing HA to build for culturing it into a DMEM-F12 medium for four weeks. The distribution of cells and ECM were assessed from histological slices stained toluidine blue and hematoxyline-eosin (HE). RESULTS: There was an increase of the number and viability of the chondrocytes during the four weeks of culture. By assessing the histological sections stained with toluidine blue and HE, we could note the definitive distribution of chondrocytes in the hydrogel, similarly to isogenous groups and territorial matrix formation. CONCLUSION: In this study, the alginate was shown to be an effective scaffold for use in chondrocytes culture, constituting an alternative for repairing joint cartilage defects.
OBJECTIVES: The aim of this study was to culture chondrocytes from knee joint cartilage of rabbits encapsulated in alginate hydrogel (HA) and to characterize the production of extracelular matrix (ECM). METHODS: Joint cartilage was obtained from rabbits' knees, three to six months old, fragmented into 1-mm pieces and submitted to enzymatic digestion. A concentration of 1x106 cells/mL were re-suspended into a 1.5% (w/v) sodium alginate solution, followed by gel formation process with CaCl2 (102 mM), allowing HA to build for culturing it into a DMEM-F12 medium for four weeks. The distribution of cells and ECM were assessed from histological slices stained toluidine blue and hematoxyline-eosin (HE). RESULTS: There was an increase of the number and viability of the chondrocytes during the four weeks of culture. By assessing the histological sections stained with toluidine blue and HE, we could note the definitive distribution of chondrocytes in the hydrogel, similarly to isogenous groups and territorial matrix formation. CONCLUSION: In this study, the alginate was shown to be an effective scaffold for use in chondrocytes culture, constituting an alternative for repairing joint cartilage defects.
Descrição
Palavras-chave
Cartilagem articular, Condrócitos, Alginato, Hidrogel, Regeneração, Engenharia tissular, Articular Cartilage, Chondrocytes, Alginate, Hydrogel, Regeneration, Tissue engineering
Como citar
Acta Ortopédica Brasileira. Sociedade Brasileira de Ortopedia e Traumatologia, v. 17, n. 4, p. 242-246, 2009.