Hongfei Qi1, Zhihong Ye1, Hailong Ren1, Nana Chen1, Qingyan Zeng1, Xianglong Wu1, Tingli Lu2. 1. Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi, People's Republic of China. 2. Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi, People's Republic of China. Electronic address: lutinglixinxin@nwpu.edu.cn.
Abstract
AIMS: The purpose of this paper was to fabricate PLLA/PCL nanofibrous scaffolds containing HAP to mimic the native bone extracellular matrix for potential applications as bone tissue engineering scaffolds materials and ultimately to help the repairing of bone defects. MATERIALS AND METHODS: PLLA (MW 200kDa), PCL (MW 80kDa), HAP, dichloromethane, N,N-dimethylformamide; α-MEM, FBS, trypsin-EDTA, penicillin G, streptomycin, β-sodium glycerophosphate, l-ascorbic acid, dexamethasone; CCK-8, Alkaline Phosphatase Assay Kit, Mouse Osteocalcin ELISA Kit, MC3T3-E1 cells. PLLA, PCL and HAP were dissolved in the solution of DCM and DMF to fabricate nanofibrous scaffolds through electrospinning. The morphology of the scaffolds was investigated with SEM, while the diameter of the fibers, pore size and water uptake of the scaffolds were tested, respectively. TGA was carried out to verify the percentage of HAP in the composite scaffolds fabricated with different HAP concentrations. Cell count kit-8 assay, alkaline phosphatase (ALP) assay, and osteocalcin assay were applied to observe the MC3T3-E1 cells proliferation, differentiation on the composite scaffolds. KEY FINDINGS: MC3T3-E1 cells were found to grow actively on the composite scaffolds based on the results of CCK-8 assay. The level of MC3T3-E1 differentiation was evaluated through the ALP activity and osteocalcin concentration, which showed higher value with HAP containing (PLLA/PCL/HAP) than that ones without (PLLA/PCL). SIGNIFICANCE: The results demonstrated that the biocomposite PLLA/PCL/HAP nanofibrous scaffold should be a promising candidate for proliferation, differentiation and mineralization of osteoblasts, and potentially can be used for bone tissue regeneration.
AIMS: The purpose of this paper was to fabricate PLLA/PCL nanofibrous scaffolds containing HAP to mimic the native bone extracellular matrix for potential applications as bone tissue engineering scaffolds materials and ultimately to help the repairing of bone defects. MATERIALS AND METHODS: PLLA (MW 200kDa), PCL (MW 80kDa), HAP, dichloromethane, N,N-dimethylformamide; α-MEM, FBS, trypsin-EDTA, penicillin G, streptomycin, β-sodium glycerophosphate, l-ascorbic acid, dexamethasone; CCK-8, Alkaline Phosphatase Assay Kit, MouseOsteocalcin ELISA Kit, MC3T3-E1 cells. PLLA, PCL and HAP were dissolved in the solution of DCM and DMF to fabricate nanofibrous scaffolds through electrospinning. The morphology of the scaffolds was investigated with SEM, while the diameter of the fibers, pore size and water uptake of the scaffolds were tested, respectively. TGA was carried out to verify the percentage of HAP in the composite scaffolds fabricated with different HAP concentrations. Cell count kit-8 assay, alkaline phosphatase (ALP) assay, and osteocalcin assay were applied to observe the MC3T3-E1 cells proliferation, differentiation on the composite scaffolds. KEY FINDINGS: MC3T3-E1 cells were found to grow actively on the composite scaffolds based on the results of CCK-8 assay. The level of MC3T3-E1 differentiation was evaluated through the ALP activity and osteocalcin concentration, which showed higher value with HAP containing (PLLA/PCL/HAP) than that ones without (PLLA/PCL). SIGNIFICANCE: The results demonstrated that the biocomposite PLLA/PCL/HAP nanofibrous scaffold should be a promising candidate for proliferation, differentiation and mineralization of osteoblasts, and potentially can be used for bone tissue regeneration.
Authors: Lilian de Siqueira; Nilza Ribeiro; Maria B A Paredes; Liliana Grenho; Cassilda Cunha-Reis; Eliandra S Trichês; Maria H Fernandes; Susana R Sousa; Fernando J Monteiro Journal: Materials (Basel) Date: 2019-11-24 Impact factor: 3.623