Mikrodalga Desteği İle Metakrilatlanmış Kappa Carrageenan Hidrojellerin 3B Baskılanması ve Kıkırdak Doku Mühendisliğinde Kullanımının Araştırılması

Abstract

This “3D Bioprinting of Microwave Assisted Methacrylated Kappa Carrageenan Hydrogels and Investigation of Their Usage in Cartilage Tissue Engineering” entitled study was supported by the The Scientific and Technological Research Council of Turkey (TUBITAK), 1002 program with project number 118M734. The aim of the presented study is development of microwave assisted methacrylated kappa carrageenan (κ-CA) polymer as a bioink with suitable properties for the 3 dimensional (3D) bioprinting, and investigation of the printed hydrogels with cartilage precursor cells (ATDC5) in cartilage tissue engineering. Within the scope of the study, the kappa carrageenan (κ-CA) precursor was methacrylated with different methacrylic anhydride (MA) ratios (4% and 12% by volume), following conventional and microwave methods. κ-CA was successfully methacrylated at low MA concentrations (4% (v/v) by microwave method to obtain a high yields of methacrylation (85 ± 9%). Then, 8% (w/v) solutions of κ-CA polymers in PBS (pH=7.4) were prepared. These solutions were treated with UV for 40 s in the presence of IRGACURE-2959® affording hydrogel structures in photo-crosslinking process, and the obtained hydrogels were fully characterized. According to the characterization results the methacrylated κ-CA polymer (12% v/v MA) by microwave process was chosen and successfully used as a bioink in 3D printing process. In the following studies, the ATDC5 cells (2 x 107 cells/mL) were mixed with the κ-CA solutions that were methacrylated with microwave and traditional method, and the result solution, in other words bioink, was 3D printed. The printed structure was cultured for 21 days in the differentiation medium. In both groups, the cells differentiated into a cartilage tissue. Scanning electron microscopy (SEM) images showed that cells exhibit spherical morphology specific to cartilage tissue. Glycosaminoglycan (GAG) content was determined by the spectrophotometric method and the presence of GAG in hydrogels was demonstrated by Safranin O / Fast Green staining. The presence of aggregan and collagen II was also demonstrated by immunocytochemical analyzes. Consequently, the 3D printed methacrylated κ-CA hydrogels including ATDC5 cells differentiated into cartilage tissue in-vitro. Thus, it is proposed that the microwave-functionalized κ-CA polymer may be considered as a suitable bioink for cartilage tissue engineering studies and microextrusion based bioprinters.