软骨终板干细胞动物体内成骨／软骨的初步研究

摘要

Objective To observe the osteogenic and/or chondrogenic differentiation potential of cartilage endplate derived stem cells ( CESCs) .Methods The cartilage endplate ( CEP) was obtained from 5 patients who underwent posterior discectomy procedure for the lumber degenerative disease.The agarose culture was used to select CESCs.The expanded CESCs were injected into the hydroxyapatite ceram-ic(HA) and incubated for 7 days.After that,the HA contained CESCs were implanted into the subcutaneous of nude mouse for 7 weeks.Then the HA was obtained and sliced for the specific stain and immunohistochemisty.The expanded CESCs were injected into the degenerated in-tervertebral disk of the rabbit for 16 weeks.Then MRI was used to detect the repair of the degenerated disc.Results The metachromasia of toluidin blue indicated that cartilage matrix was formed in the vacant space of HA.Mallory stain indicated that there was bone matrix was formed.The results of immunohistochemistry indicated that there was more collageⅠand collageⅩ,and a small quantity of collage Ⅱ.MRI results indicated that the CESCs could be induced into the chondrocyte-like cells in the rabbit intervertebral disk.Conclusion The CESCs could be induced into the bone/cartilage in vivo.%目的：观察软骨终板干细胞在动物体内成骨／软骨的能力。方法临床获取腰椎退变椎间盘软骨终板，采用琼脂糖筛选软骨终板干细胞后进行传代扩增，扩增后的软骨终板干细胞与羟基磷灰石复合后植入裸鼠皮下，7周后处死裸鼠取出复合物，分别进行特殊染色和免疫组化染色分析。同时，将扩增的干细胞注入兔子退变椎间盘中，16周后行MRI检测，观察退变椎间盘修复情况。结果软骨终板干细胞羟基磷灰石复合物切片经甲苯胺蓝染色后有异染性，提示有软骨基质形成；同时Mallory染色显示羟基磷灰石空隙中有蓝染存在，说明有成熟的骨基质形成。免疫组织化学结果提示，所形成的基质主要为Ⅰ型和Ⅹ型胶原，同时有少量Ⅱ型胶原表达。 MRI结果提示植入退变椎间盘的终板干细胞在动物体内可以分化为类软骨样细胞，从而修复退变椎间盘。结论软骨终板干细胞可以在动物体内向成骨／软骨方向分化。