基于细胞外基质来源的组织工程椎间盘双相支架裸鼠皮下异位构建椎间盘

发布时间：2018-10-30 07:43

【摘要】：[目的]探讨椎间盘细胞和细胞外基质来源的一体化纤维环-髓核双相支架在裸鼠体内异位构建组织工程一体化椎间盘的可行性,并采用PKH26荧光标记和小动物活体荧光成像系统无创评估组织工程化细胞-支架复合体在体内生长情况。[方法]纤维环细胞和髓核细胞分别标记PKH26荧光,分别接种入细胞外基质来源的一体化支架不同相中,扫描电镜、Dead/Live荧光染色观察细胞粘附及活性,植入裸鼠背部皮下,6周后利用分子小动物活体荧光成像系统评价组织工程化组织在裸鼠体内生长情况,取材进行荧光显微镜下观察、组织学染色。[结果]扫描电镜观察细胞粘附在双相支架上且周围有基质分泌,Dead/Live染色示细胞在双相支架上活性良好;6周后,活体荧光成像显示椎间盘细胞在支架内生长良好,从髓核往纤维环荧光强度减弱,细胞支架复合体在裸鼠体内形成椎间盘样组织,HE、番红O染色、甲苯胺蓝染色阳性。[结论]天然骨基质明胶和软骨基质来源的一体化纤维环-髓核支架复合椎间盘细胞能够在裸鼠皮下异位构建椎间盘样组织。
[Abstract]:[objective] to investigate the feasibility of constructing an integrated intervertebral disc in nude mice with intervertebral disc cells and extracellular matrix (ECM) derived fibro-annulus pulposus biphasic scaffolds in vivo. The growth of tissue-engineered cell-scaffold complexes in vivo was evaluated by PKH26 fluorescence labeling and small animal in vivo fluorescence imaging system. [methods] Fibroannular cells and nucleus pulposus cells were labeled with PKH26 fluorescence, respectively, and inoculated into different phases of integrated scaffold derived from extracellular matrix. Scanning electron microscopy and Dead/Live fluorescence staining were used to observe cell adhesion and activity. After 6 weeks, the growth of tissue engineered tissue in nude mice was evaluated by fluorescence imaging system of small molecular animals in vivo. The samples were observed under fluorescence microscope and stained by histology. [results] the cell adhesion to the biphasic scaffold was observed by scanning electron microscope (SEM) and there was matrix secretion around the scaffold. Dead/Live staining showed that the cell was active on the biphasic scaffold. After 6 weeks, the in vivo fluorescence imaging showed that the intervertebral disc cells grew well in the scaffold, the fluorescence intensity from the nucleus pulposus to the fibrous ring decreased, the scaffold complex formed disklike tissue in the nude mice, and HE, staining. Toluidine blue staining was positive. Conclusion the intervertebral disc cells derived from natural bone matrix gelatin and cartilage matrix can construct intervertebral disc-like tissue in nude mice subcutaneously.
【作者单位】：天津医科大学研究生院;天津市天津医院;天津理工大学机械工程学院;
【基金】：国家自然科学基金课题资助项目(编号:81272046,31000432,31300798) 中国博士后科学基金项目(编号:2011M500530,2012T50221) 天津市卫生局科技基金项目(编号:2013KR16)
【分类号】：R318.08

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