Three dimensional（3D） bioprinting, which involves depositing bioinks（mixed biomaterials） layer by layer to form computer-aided designs, is an ideal method for fabricating complex 3D biological structures. However,...Three dimensional（3D） bioprinting, which involves depositing bioinks（mixed biomaterials） layer by layer to form computer-aided designs, is an ideal method for fabricating complex 3D biological structures. However, it remains challenging to prepare biomaterials with micro-nanostructures that accurately mimic the nanostructural features of natural tissues. A novel nanotechnological tool, electrospinning, permits the processing and modification of proper nanoscale biomaterials to enhance neural cell adhesion, migration, proliferation, differentiation, and subsequent nerve regeneration. The composite scaffold was prepared by combining 3D bioprinting with subsequent electrochemical deposition of polypyrrole and electrospinning of silk fibroin to form a composite polypyrrole/silk fibroin scaffold. Fourier transform infrared spectroscopy was used to analyze scaffold composition. The surface morphology of the scaffold was observed by light microscopy and scanning electron microscopy. A digital multimeter was used to measure the resistivity of prepared scaffolds. Light microscopy was applied to observe the surface morphology of scaffolds immersed in water or Dulbecco＇s Modified Eagle＇s Medium at 37℃ for 30 days to assess stability. Results showed characteristic peaks of polypyrrole and silk fibroin in the synthesized conductive polypyrrole/silk fibroin scaffold, as well as the structure of the electrospun nanofiber layer on the surface. The electrical conductivity was 1 × 10^-5–1 × 10^-3 S/cm, while stability was 66.67%. A 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay was employed to measure scaffold cytotoxicity in vitro. Fluorescence microscopy was used to observe Ed U-labeled Schwann cells to quantify cell proliferation. Immunohistochemistry was utilized to detect S100β immunoreactivity, while scanning electron microscopy was applied to observe the morphology of adherent Schwann cells. Results demonstrated that the polypyrrole/silk fibroin scaffold was not cytotoxic and 展开更多
[目的]通过比较常用的M-CSF与L929细胞培养上清两种诱导方式获得骨髓巨噬细胞(BMDM)的形态及基本生物学功能,为选择制备BMDM的方法提供实验依据。[方法]用两种方式诱导小鼠骨髓获取BMDM,然后分别利用Zeiss荧光显微镜和流式细胞仪比较BMD...[目的]通过比较常用的M-CSF与L929细胞培养上清两种诱导方式获得骨髓巨噬细胞(BMDM)的形态及基本生物学功能,为选择制备BMDM的方法提供实验依据。[方法]用两种方式诱导小鼠骨髓获取BMDM,然后分别利用Zeiss荧光显微镜和流式细胞仪比较BMDM的细胞形态及纯度;通过吞噬实验、杀菌实验以及液相芯片技术(liquid chip)比较两组BMDM的生物学功能;同时将上述结果与体内分化成熟的腹腔巨噬细胞相比较。[结果]两组诱导分化成熟的BMDM均呈现不规则形或梭形;BMDM的纯度分别为M-CSF组98.6%,L929上清组99.6%;腹腔细胞贴壁后巨噬细胞的纯度是98.8%;吞噬实验、杀菌实验两组之间及其与腹腔巨噬细胞相比无显著差异;炎症因子释放水平检测中3组细胞存在IL-6或TNF-α某些时间点释放水平差异。[结论]M-CS与L929上清诱导方式所获得BMDM的形态相似,纯度相近(98.6%比99.6%),吞噬细菌能力无显著统计学差异(21.31±5.83比26.10±6.11,t=-0.745,P>0.05),杀菌能力无显著差异(杀菌30min:36.41%±4.21%比39.53%±6.75%;60min:44.35%±6.75%比45.27%±1.96%,两者均P>0.05),但是炎症因子释放的某些时间点有差异(TNF-α的释放在LPS刺激后24 h M-CSF组高于L929组:549.92±412比271.47±432,t=-0.34,P<0.05),应根据实验目的选择获取BMDM的诱导方式。展开更多
The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios.The water contents of the hydrogels were in the range of 50%～80%(wt).The attachment and growth of fibroblast cells(L929...The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios.The water contents of the hydrogels were in the range of 50%～80%(wt).The attachment and growth of fibroblast cells(L929) on the hydrogels were studied.The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.展开更多
基金supported by the National Natural Science Foundation of China,No.81671823,81701835a grant from the National Key Research and Development Program of China,No.2016YFC1101603a grant from the Natural Science Research Program of Nantong of China,No.MS12016056
文摘Three dimensional（3D） bioprinting, which involves depositing bioinks（mixed biomaterials） layer by layer to form computer-aided designs, is an ideal method for fabricating complex 3D biological structures. However, it remains challenging to prepare biomaterials with micro-nanostructures that accurately mimic the nanostructural features of natural tissues. A novel nanotechnological tool, electrospinning, permits the processing and modification of proper nanoscale biomaterials to enhance neural cell adhesion, migration, proliferation, differentiation, and subsequent nerve regeneration. The composite scaffold was prepared by combining 3D bioprinting with subsequent electrochemical deposition of polypyrrole and electrospinning of silk fibroin to form a composite polypyrrole/silk fibroin scaffold. Fourier transform infrared spectroscopy was used to analyze scaffold composition. The surface morphology of the scaffold was observed by light microscopy and scanning electron microscopy. A digital multimeter was used to measure the resistivity of prepared scaffolds. Light microscopy was applied to observe the surface morphology of scaffolds immersed in water or Dulbecco＇s Modified Eagle＇s Medium at 37℃ for 30 days to assess stability. Results showed characteristic peaks of polypyrrole and silk fibroin in the synthesized conductive polypyrrole/silk fibroin scaffold, as well as the structure of the electrospun nanofiber layer on the surface. The electrical conductivity was 1 × 10^-5–1 × 10^-3 S/cm, while stability was 66.67%. A 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay was employed to measure scaffold cytotoxicity in vitro. Fluorescence microscopy was used to observe Ed U-labeled Schwann cells to quantify cell proliferation. Immunohistochemistry was utilized to detect S100β immunoreactivity, while scanning electron microscopy was applied to observe the morphology of adherent Schwann cells. Results demonstrated that the polypyrrole/silk fibroin scaffold was not cytotoxic and
文摘[目的]通过比较常用的M-CSF与L929细胞培养上清两种诱导方式获得骨髓巨噬细胞(BMDM)的形态及基本生物学功能,为选择制备BMDM的方法提供实验依据。[方法]用两种方式诱导小鼠骨髓获取BMDM,然后分别利用Zeiss荧光显微镜和流式细胞仪比较BMDM的细胞形态及纯度;通过吞噬实验、杀菌实验以及液相芯片技术(liquid chip)比较两组BMDM的生物学功能;同时将上述结果与体内分化成熟的腹腔巨噬细胞相比较。[结果]两组诱导分化成熟的BMDM均呈现不规则形或梭形;BMDM的纯度分别为M-CSF组98.6%,L929上清组99.6%;腹腔细胞贴壁后巨噬细胞的纯度是98.8%;吞噬实验、杀菌实验两组之间及其与腹腔巨噬细胞相比无显著差异;炎症因子释放水平检测中3组细胞存在IL-6或TNF-α某些时间点释放水平差异。[结论]M-CS与L929上清诱导方式所获得BMDM的形态相似,纯度相近(98.6%比99.6%),吞噬细菌能力无显著统计学差异(21.31±5.83比26.10±6.11,t=-0.745,P>0.05),杀菌能力无显著差异(杀菌30min:36.41%±4.21%比39.53%±6.75%;60min:44.35%±6.75%比45.27%±1.96%,两者均P>0.05),但是炎症因子释放的某些时间点有差异(TNF-α的释放在LPS刺激后24 h M-CSF组高于L929组:549.92±412比271.47±432,t=-0.34,P<0.05),应根据实验目的选择获取BMDM的诱导方式。
基金国家重点基础研究发展规划 ( 973)项目基金资助 (No .G19990 6 470 5 )~~
文摘The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios.The water contents of the hydrogels were in the range of 50%～80%(wt).The attachment and growth of fibroblast cells(L929) on the hydrogels were studied.The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.
