The assessment of natural scaffolds ability in chondrogenic differentiation of human adipose-derived mesenchymal stem cells

Abstract

ABSTRACT:The ability of cartilage to repair damage is limited due to lack of blood vessels and low cell density. Recently, tissue engineering is considerably preferred to other treatments as a way to solve this problem. Regardless of cell sources, one of the crucial factors in tissue engineering is to select an appropriate scaffold, which is essential for healing and renewal procedure of tissues in vivo and in vitro. Mesenchymal stem cells (MSCs) were isolated from adipose tissue in liposuction surgeries by use of collagenase enzyme. After verification by flow cytometry methods, adipose-derived mesenchymal stem cells (ADMSCs) were embedded into alginate and agarose scaffolds, separately; and then they were cultured in chondrogenic medium for 3 weeks. The ability of alginate and agarose scaffolds was assessed by use of MTT assay and histological analysis. In addition, analysis of chondrogenic genes expression by Real-time PCR was done. The obtained data were analyzed statistically by means of SPSS software. There was a significant difference between alginate and agarose groups in maintaining cells viable but, about chondrogenic differentiation analyzed by use of real-time PCR, statistical analysis has shown a significant difference in expression of aggrecan (as a chondrocyte-specific gene) and collagen II (as an chondrocyte-specific gene) between cell/alginate and cell/agarose and MSCs (p<0.05). Chondrocyte differentiation of cells was verified by histological analysis. Alginate scaffold can provide a suitable environment for chondrogenic differentiation of adipose derived mesenchymal stem cells.

Mesenchymal stem cells as a valuable agent in osteoarthritis treatment

“…The application of conventional treatment methods, such as pain medication and physical therapy techniques, could temporarily alleviate pain, but these methods do not have the ability to recover the primary function of the cartilage (3). Additionally, many difficulties are encountered in autologous chondrocyte implantation for cartilage regeneration, such as donor site damage and cell limited life span after transplantation (4,5). These problems make new therapeutic methods, such as regenerative medicine, to repair the damaged cartilage tissue an exciting prospect (6).…”

Electrospun nanofibrous alginate sulfate scaffolds promote mesenchymal stem cells differentiation to chondrocytes

“…Several in vitro and in vivo researches have published MSC‐scaffold‐supported chondrogenic differentiation, such as collagen, polyethersulfone, gelatin, hyaluronic acid, alginate. Also, research of Sonomoto et al confirmed that MSCs scaffold heightens cartilage repair with increased SOX9 and COL10A1 expression 18–21 …”

“…Also, research of Sonomoto et al confirmed that MSCs scaffold heightens cartilage repair with increased SOX9 and COL10A1 expression. [18][19][20][21] Alginate is a promising natural hydrophilic polysaccharide and considered as one of the excellent biomaterials for cartilage tissue engineering due to its favorable biocompatibility, accessibility, low toxicity, useful structural feature, biodegradability, and cost-effective. However, neat alginate as a bioinert material shows limited interactions with biomolecules of ECM and the cells.…”

Filmes de Langmuir, nanopartículas e biomembranas para estudos de biomineralização óssea: do modelo biomiméticos às aplicações biotecnológicas

“…297 Os mecanismos de sinalização intracelular desencadeados durante a diferenciação condrogênica e osteogênica foram revisados. [298][299][300][301][302] Vários deles são mediados por membros da família de proteínas quinase ativada por mitógenos (MAP) e proteína quinase extracelular regulada (ERK), que ativam o fator de transcrição Cbfα-1, induzindo assim a regulação do genes específicos dos ossos. 303 Os scaffolds 3D baseados em COL1 aumentaram a ativação dessas vias de sinalização em comparação com culturas 2D, que também afetou o fenótipo celular.…”

“…[350][351][352][353] −Carr é um dos três tipos mais importantes de carragenana, que exibe o menor grau de sulfatação com um grupo sulfato nas unidades da cadeia de galactose/anidrogalactose. 298,332 Na presença de cátions específicos, esse polissacarídeo se organiza em uma estrutura helicoidal, como a do COL. As propriedades do gel, como tempo de gelificação, temperatura de transição sol-gel, dureza e sinérese, dependem do arranjo supramolecular das hélices relacionadas à estrutura do polissacarídeo e sua agregação, geralmente promovida por cátions de metal mono e divalentes. 331 Devido à sua biocompatibilidade, hidrofilicidade e capacidade de estimular a adesão e proliferação celular, os hidrogéis à base de −Carr podem ser aplicados em sistemas carregados de células para a regeneração de cartilagem e ossos.…”