纳米硅取代羟基磷灰石/脱细胞骨基质复合材料的研究

发布时间：2022-01-22 12:07

　　本论文从牛、猪，大鼠和人的骨组织中提取脱钙/脱细胞骨基质（DBM/dDBM），和羟基磷灰石/纳米晶硅取代羟基磷灰石（HA/nSiHA）复合，制备了复合支架材料，用细胞实验初步评估了所得的支架材料，研究了骨传导和骨诱导性能，这些研究结果对同种异体移植/异种器官移植等研究领域具有重要的意义。论文分析了以下因素对骨髓间充质干细胞培养系统的影响：物种间的生物相容性、脱细胞过程、 pH值、DBM/dDBM粉与磷灰石的重量比、细胞分化培养基。首先，论文对DBM/dDBM的制备过程进行了优化，得到了较好的脱细胞水平：50ng dsDNA/mg dDBM。制备支架材料的HA和nSiHA粉末通过化学沉淀法合成。所得支架和培养后支架通过SEM、EDX、FT-IR、组织学和免疫组化染色进行了表征。通过压片的方法，制备了不同重量含量的DBM/dDBM与HA/nSiHA复合支架：100%、40%、20%、10%和0%。将所得支架在蒸馏水和PBS中浸泡21和28天，检验了其稳定性和pH值变化。将支架进行细胞培养，研究了其细胞增殖和分化的结果。将骨髓间充质干细胞在支架上培养24小时后，用细胞毒性试剂盒和SEM分析了... 

【文章来源】：华中科技大学湖北省 211工程院校 985工程院校 教育部直属院校

【文章页数】：88 页

【学位级别】：硕士

【文章目录】：
摘要
Abstract
Contents
1 INTRODUCTION
    1.1 Aim
    1.2 Rationale
    1.3 The key issue of optimization
2 MATERIALS AND METHODS
    2.1 Bone preparation
    2.2 Demineralization
    2.3 Decellularization
    2.4 nSiHA and HA production
    2.5 DBM/dDBM morphology characterization - Evidence of having osteogenic and osteoconductive components - Scanning electron microscopy (SEM)
    2.6 DBM/dDBM functional groups characterization - Evidence of having osteogenic and osteoconductive components - Fourier transform infrared (FT-IR) spectroscopy)
    2.7 DBM/dDBM cellular characterization - Evidence of having osteogenic and osteoconductive components - Immunohistological & histological staining
    2.8 Incorporation of DBM/dDBM and HA/nSiHA
    2.9 Scaffold fabrication and substrate distribution
    2.10 HA/nSiHA characterization
    2.11 Local pH
    2.12 Mass loss (lyophilization) and SEM (degradation)
    2.13 Sterilization
    2.14 Cell culture
    2.15 Cell seeding and culture in vitro
    2.16 Cytotoxicity - Cck8 assay
    2.17 Cytotoxicity - Scanning electron microcopy (SEM)
    2.18 Osteogenic induction
    2.19 Differentiation - Characterization of osteogenic differentiation using morphological features - Optical microscopy & immunohistological staining (DAPI)
    2.20 Differentiation - Alkaline phosphatase activity (ALP)
3 EVALUATION PLAN
    3.1 Lyophilization and decellularization
    3.2 pH
    3.3 Cytotoxicity
    3.4 Differentiation
4 RESULTS
    4.1 DBM/dDBM production and lyophilization
    4.2 nSiHA and HA production
    4.3 DBM/dDBM morphology characterization - Evidence of having osteogenic and osteoconductive components - Scanning electron microscopy (SEM)
    4.4 DBM/dDBM functional groups characterization - Evidence of having osteogenic and osteoconductive components - Fourier transform infrared (FT-IR) spectroscopy
    4.5 DBM/dDBM cellular characterization - Evidence of having osteogenic and osteoconductive components
    4.6 HA and nSiHA characterization – SEM and EDX
    4.7 Local pH
    4.8 Degradation - SEM
    4.9 Cytotoxicity
    4.10 Differentiation
5 DISCUSSION
6 CONCLUSION
Acknowledgements
REFERENCES



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