RESUMO
Cold Atmospheric Plasma (CAP) is used as a promising method in surface modification for immobilization of chondroitin sulfate functional biomacromolecules on PCL nanofibrous substrates for cartilage tissue engineering. The GAG-grafted scaffolds are able to successfully support the attachment and proliferation of mesenchymal stem cells (MSCs). The seeded scaffolds show the chondro-differentiation of MSCs during a 21-days cell culture in a non-differential medium. Expression of SOX9, Collagen10 and Collagen2 proved the chondro-inductive effect of GAG-grafted scaffolds. Besides, no external chondro-genic differential agent was used in the differentiation of MSCs to chondrocyte. The cells passed the last phase of chondrogenesis after 14â¯days of incubation. Thus, the GAG-fabricated fibrous scaffolds using CAP are potential candidates for cartilage tissue engineering.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Condrogênese/efeitos dos fármacos , Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanofibras/química , Poliésteres/química , Condrócitos/citologia , Condrócitos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Teste de Materiais , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismoRESUMO
Poly(Æ-caprolactone) (PCL) is a biocompatible polymer with a high potential to be used in tissue engineering especially in tight tissues. In the current study, cold atmospheric plasma (CAP) is used as a promising method for immobilization of gelatin as a functional biomacromolecule on PCL nanofibrous substrates. The CAP surface modification leads to oxidation of chemical groups existing on the PCL surface without doing any damage to the bulk properties of biomaterials for gelatin biomacromolecule grafting. The water contact angle (WCA) of the CAP-treated surface and gelatin-grafted PCL using CAP indicates an effective increment in the hydrophilicity of the PCL surface. Also to achieve the highest levels of gelatin grafting on the PCL surface, two different grafting methods and gelatin concentration diversity are utilized in the grafting process. The immobilization of gelatin biomacromolecules onto the CAP surface-modified PCL nanofibers is investigated using scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR). The gelatin-modified PCL substrates revealed uniform nanofibrous morphology with increased average fiber diameter. The results of FTIR spectra, including hydroxyl groups, NH groups, and amide II of gelatin-grafting peaks, confirm the gelatin immobilization on the surface of nanofibers. The metabolic activity of cultured mesenchymal stem cells (MSCs) on the surface-modified scaffolds is evaluated using MTT analysis (P ≤ 0.05). The results of metabolic activity and also SEM and DAPI staining observations indicate proper attachment on the surface and viability for MSCs on the surface-immobilized nanofibrous scaffolds. Therefore, CAP treatment would be an effective method for biomacromolecule immobilization on nanofibers towards the enhancement of cell behavior.