椎间盘器官培养模型的建立及临床意义

目的 建立类似于体内生理环境的椎间盘器官培养模型并评估其临床意义。方法 研究材料取白10周龄小鼠的腰椎脊柱功能单位,每个脊柱功能单位由椎间盘和两端的部分椎体组成。将小鼠的腰椎脊柱功能单位在培养液和不同浓度的布比卡凶溶液中,经过不同时段的培养,通过组织学染色和MTT方法 检测椎间盘的组织学变化和细胞活性。结果 小鼠椎问盘的组织学特点和细胞活性在培养液中维持4周无明显变化。0.25％布比卡因中培养1h导致25％椎间盘细胞死亡,而浓度为0.5％相同时间培养死亡细胞比例上升至60％。结论 小鼠椎间盘经体外器官培养4周,维持了组织结构和细胞功能的完整性。随着布比卡因作用浓度的增加,椎间盘的细胞活性明显降低。

Development of an intervertebral disc organ culture model and its clinical significance

Objective To develop an in vivo intervertebral disc organ culture model for a physiological environment and evaluate its clinical significance. Methods Murine functional spine units (FSUs) were isolated from 10-week-old mouse lumbar spines. FSUs consisted of two vertebrae surrounding one disc. Murine FSUs were cultured in medium and different concentrations of bupivacaine for different periods. Histological change and cell viability within intervertebral disc tissue were assessed by histological staining and MTT (3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. Results Murine disc organs cultured for up to 4 weeks showed minimal changes in tissue histology and cell viability. A 1 -hour incubation in 0.25％ bupivacaine resulted in about 25％ cell death while 0.5％ bupivacaine exposure yielded 60％ cell death over the same time. Conclusion Murine intervertebral disc maintains the integrity of tissue structures and cell functions for an ex vivo 4-week culture. And the exposure to bupivacaine dramatically decreases cell viability in a dose-dependent manner.