Cultura de condrócitos em arcabouço tridimensional: hidrogel de alginato

Abstract

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.

Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules

“…Similar results to the present study were observed using 10 4 cells per microcapsules, through maintenance and cell proliferation for up to 4 weeks. With similar methodology to the present study, using 10 4 cells per microcapsules, was observed through maintenance and cell proliferation for up to 4 weeks [ 30 ]. Studies that performed the encapsulation of MSC from human bone marrow, at the concentration of sixty thousand cells per alginate microcapsule, reported that viability remained high (> 90%) over 4 weeks and the duration of culture in vitro had no effect on the cell viability [ 37 ].…”

“…A study using six different types of comercial alginate demostrated that type may influence chondrocyte culture upon finding variation in their pH, viscosity or n-gururonic acid-manuronic acid ratio [ 39 ]. Researchers confirmed that variations alginate concentration influence the cellular metabolism and diffusion of the necessary components of the culture medium, recommending the use of 1.5% (w/v) [ 30 ]. Needle diameter may interfere with cell viability, concluding that larger gauge (19G) needles for cell implantation reduce the loss of stress-induced viability compared to smaller (21G and 23G) needle gauges [ 40 ].…”

“…The gelatinization solution was discarded, and the microcapsules were washed 3 times in 5 vol. 0.15 M NaCl [ 30 ]. The microcapsules were cultured in Knockout™ DMEM medium.…”

3D Cell Culture in Alginate Hydrogels

“…Cells producing the anti‑angiogenic protein endostatin have been encapsulated in ultrapure alginate and implanted into the brains of dogs being treated for spontaneous brain cancer [84,85,86]. Many other cell types have been immobilized in alginate such as adipose-derived stem cells [87,88], mesenchymal stem cells [89,90], and chondrocytes [91,92]. …”

Light-emitting diode effects on combined decellularization of tracheae. A novel approach to obtain biological scaffolds

“…Even though a cell is usually able to overcome small disturbances of the redox balance, larger-scale disturbances may be lethal to the cells [17][18][19] , such the apoptosis caused by oxidative stress resulting from LED irradiation 17 . Our research group has been interested in suitable scaffolds for tissue engineering for a long time 12,20,21 . This paper describes different techniques of cell removal from tracheae as a proposed three dimensional scaffold, by associating traditional approaches with the novel LED proposal.…”