本論文從牛、豬，大鼠和人的骨組織中提取脫鈣/脫細(xì)胞骨基質(zhì)（DBM/dDBM），和羥基磷灰石/納米晶硅取代羥基磷灰石（HA/nSiHA）復(fù)合，制備了復(fù)合支架材料，用細(xì)胞實(shí)驗(yàn)初步評(píng)估了所得的支架材料，研究了骨傳導(dǎo)和骨誘導(dǎo)性能，這些研究結(jié)果對(duì)同種異體移植/異種器官移植等研究領(lǐng)域具有重要的意義。論文分析了以下因素對(duì)骨髓間充質(zhì)干細(xì)胞培養(yǎng)系統(tǒng)的影響：物種間的生物相容性、脫細(xì)胞過(guò)程、 pH值、DBM/dDBM粉與磷灰石的重量比、細(xì)胞分化培養(yǎng)基。首先，論文對(duì)DBM/dDBM的制備過(guò)程進(jìn)行了優(yōu)化，得到了較好的脫細(xì)胞水平：50ng dsDNA/mg dDBM。制備支架材料的HA和nSiHA粉末通過(guò)化學(xué)沉淀法合成。所得支架和培養(yǎng)后支架通過(guò)SEM、EDX、FT-IR、組織學(xué)和免疫組化染色進(jìn)行了表征。通過(guò)壓片的方法，制備了不同重量含量的DBM/dDBM與HA/nSiHA復(fù)合支架：100%、40%、20%、10%和0%。將所得支架在蒸餾水和PBS中浸泡21和28天，檢驗(yàn)了其穩(wěn)定性和pH值變化。將支架進(jìn)行細(xì)胞培養(yǎng)，研究了其細(xì)胞增殖和分化的結(jié)果。將骨髓間充質(zhì)干細(xì)胞在支架上培養(yǎng)24小時(shí)后，用細(xì)胞毒性試劑盒和SEM分析了... 

【文章來(lái)源】：華中科技大學(xué)湖北省 211工程院校 985工程院校 教育部直屬院校

【文章頁(yè)數(shù)】：88 頁(yè)

【學(xué)位級(jí)別】：碩士

【文章目錄】：
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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