Xie Jian, Tong Pei-Jian, Xiao Lu-Wei, Jin Hong-Ting, Wu Cheng-Liang, Shan Le-Tian, Mao Qiang, Pan Jia-Fei
Zhejiang University of Traditional Chinese Medicine, Hangzhou 310053, Zhejiang, China.
Zhongguo Gu Shang. 2013 Jun;26(6):481-5.
To compare biological characteristics between nucleus pulposus and annulus fibrosus cells in vitro model.
Five New Zealand white rabbits (2 to 3 kg, either gender) were isolated nucleus pulposus and annulus fibrosus under sterilized condition, then cultured in nutrient solution with 15% FBS and DMEM/F12 (1:1) by enzyme digestion combined with tissue block method. When 90% cells fused, subcultring were performed. Cell morphology were observed by inverted phase contrast microscope, cell viability were detected by trypan blue staining, histological were observed by a toluidine blue and HE staining, cell proliferation were tested by MTT method, then the cell morphology, viability, proliferation between nucleus pulposus and annulus fibrosus were compared.
There were no obviously differences between nucleus pulposus and annulus fibrosus in original and the first strain. Physalides were appeared in annulus fibrosus on the second generation. The strapping time was later, and activity was lower in nucleus pulposus than annulus fibrosus. The growth of cell proliferation in nucleus pulposus was lower than annulus fibrosus from the ninth day.
The cell activity in annulus fibrosus is higher than nucleus pulposus. Digenerative disc disease may caused by recession of nucleus pulposus,local biomechnical changes, furether caused structure change and function loss of annulus fibrosus.
比较体外模型中髓核细胞和纤维环细胞的生物学特性。
选取5只新西兰白兔（2至3千克，雌雄不限），在无菌条件下分离髓核和纤维环，然后通过酶消化结合组织块法，在含15%胎牛血清和DMEM/F12（1:1）的营养液中培养。当细胞融合达90%时进行传代培养。通过倒置相差显微镜观察细胞形态，用台盼蓝染色检测细胞活力，用甲苯胺蓝和苏木精-伊红染色观察组织学变化，用MTT法检测细胞增殖，然后比较髓核和纤维环之间的细胞形态、活力及增殖情况。
原代及第一代时髓核和纤维环之间无明显差异。第二代纤维环出现成纤维细胞。髓核的贴壁时间较晚，活性低于纤维环。从第9天起，髓核细胞增殖生长低于纤维环。
纤维环细胞活性高于髓核。椎间盘退变疾病可能由髓核退变、局部生物力学改变引起，进而导致纤维环结构改变和功能丧失。
