| Objective In this research,we try to investigate the applications of chitosan (CHT)/polycaprolactone (PCL) blend scaffolds in bio-engineering, and then to make a comparison of the capability of three different types of CHT/PCL blend scaffolds to promote cartilage repair, and analysis of the internal mechanism of the performance.Methods The present experiment using three different types of chitosan (CHT)/polycaprolactone (PCL) blend scaffolds:100:0(100CHT),75:25(75CHT) and50:50(50CHT)wt.%CHT/PCL. CHT and PCL was dissolved in a solution of acetic acid of different concentrations, respectively, and in accordance with a predetermined weight ratio of mixture of two solution, and adding a certain amount of salt particles, mixing, heating and then cast into membrane onto a Teflon dish, followed by removing the salt particles in a5.0wt%NaOH solution. After that, they were dehydrated and totally dried at ambient temperature. The three different types of blend scaffolds were obtained. Scanning electron microscopy (SEM) analysis revealed an adequate porosity, pore size and interconnectivity for cartilage tissue engineering. Chondrocytes were separated from SD rats and cultured, using type Ⅱ collagen immunohistochemical staining to identify the cell types. The second generation chondrocyte cell suspension was prepared and inoculated into the three-dimensional scaffolds, cultured in a cell incubator. At day3,7,14,21of cuiture we measured the total number of viable cells and its metabolic activity in the scaffolds, and detected the formation of the extracellular matrix (ECM). Besides, section staining and scanning electron microscopy were peformed to observed cell line state, and then executed comparative analysis of capability of promoting cartilage repair among each groups.Results The experiments showed that cells attached, survival, proliferated and were metabolically active over all scaffolds formations. Cartilage extracellular matrix (ECM) formation was observed in all formulations. Total determination of the proliferation cell revealed that75CHT had the largest number of new cartilage cells, and formed the most amount of ECM, significantly higher than the50CHT and100CHT. These results were statistically significant (P<0.05). Scanning electron microscopy (SEM) showed uniform distribution of new cartilage cells in the surface of the scaffolds occurred.Conclusions The75CHT scaffolds supported the most new cartilage tissue formation, and maintained chondrocytes physiological characteristics.It may be more suitable for the repair of cartilage tissue engineering. |
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