丝素蛋白/纳米羟基磷灰石/淫羊藿苷可促进移植骨髓间充质干细胞增殖分化为髓核样细胞  

Silk fibroin/nano hydroxyapatite composite combined with icariin can promote the proliferation and differentiation of bone marrow mesenchymal stem cells into nucleus pulposus like cells

作　　者：郭晓鹏[1] 刘莹松[1] 尚晖[1] Guo Xiaopeng;Liu Yingsong;Shang Hui(Shiyan Taihe Hospital,Shiyan 442000,Hubei Province,China)

机构地区：[1]十堰市太和医院,湖北省十堰市442000

出　　处：《中国组织工程研究》2022年第22期3528-3534,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:近年来利用组织工程技术修复椎间盘退变成为脊柱外科领域的研究热点。目的:对比丝素蛋白/纳米羟基磷灰石/淫羊藿苷与丝素蛋白/纳米羟基磷灰石复合材料在椎间盘退变修复中的效果。方法:采用物理共混法与致孔剂制备丝素蛋白/纳米羟基磷灰石水凝胶,将水凝胶直接与淫羊藿苷物理混合制备含淫羊藿苷(10 mmol/L)的水凝胶。采用髓核抽吸法建立新西兰大白兔椎间盘退变动物模型,将造模成功的36只兔随机分3组,每组12只:模型组采用微量注射器向椎间盘内注射PBS,对照组注射丝素蛋白/羟基磷灰石水凝胶与骨髓间充质干细胞悬液,实验组注射丝素蛋白/羟基磷灰石水/淫羊藿苷凝胶与骨髓间充质干细胞悬液。正常组(n=12)不建模也不进行任何处理。术后4,8,12周进行X射线片、MRI检查,术后12周进行组织学(苏木精-伊红染色与甲苯胺蓝染色)、RT-PCR检测(Ⅱ型胶原、蛋白聚糖、Sox9 mRNA的表达)与Western blotting检测(Ⅱ型胶原、蛋白聚糖、Sox9蛋白的表达)。结果与结论:(1)X射线片显示,术后4,8,12周时,模型组椎间盘高度逐渐下降,对照组、实验组椎间盘高度逐渐增加,其中实验组兔椎间盘高度的恢复好于对照组;(2)MRI检查显示,术后4,8,12周时,模型组兔椎间盘T2加权像信号值呈逐渐降低趋势,对照组与实验组逐渐升高,其中以实验组升高更明显;(3)苏木精-伊红染色显示,模型组髓核组织破坏较明显,髓核细胞显著减少;对照组髓核组织欠完整,中央髓核区可见少量髓核样细胞增生;实验组髓核组织较完整,中央髓核区可见大量髓核样细胞增生;(4)甲苯胺蓝染色显示,模型组髓核无着色,对照组中度染色,实验组髓深染;(5)RT-PCR与Western blotting检测显示,模型组Ⅱ型胶原、蛋白聚糖、Sox9的mRNA及蛋白表达水平低于正常组(P<0.05),对照组与实验组的Ⅱ型胶原、蛋白聚糖、Sox9的mRNA及蛋白表达水平BACKGROUND:In recent years,using tissue engineering technology to repair intervertebral disc degeneration has become a research hotspot in the field of spinal surgery.OBJECTIVE:To compare the effect of silk fibroin/nano hydroxyapatite composite material combined with icariin and silk fibroin/nano hydroxyapatite composite in the repair of intervertebral disc degeneration.METHODS:Silk fibroin/nano hydroxyapatite hydrogel was prepared by physical blending and porogen.The hydrogel was directly physically mixed with icariin to prepare a hydrogel containing icariin(10 mmol/L).Animal models of intervertebral disc degeneration in New Zealand white rabbits were established by nucleus pulposus aspiration method.Thirty-six rabbits were randomly divided into 3 groups,with 12 in each group.The model group was injected with PBS using microinjector into intervertebral disc.The control group was injected with silk fibroin/hydroxyapatite hydrogel and bone marrow mesenchymal stem cell suspension.The experimental group was injected with fibroin/hydroxyapatite hydrogel containing icariin and bone marrow mesenchymal stem cell suspension.The normal group(n=12)was not modeled and did not receive any treatment.X-ray and MRI examinations were performed 4,8,and 12 weeks after surgery.Histology(hematoxylin-eosin staining and toluidine blue staining),RT-PCR detection(typeⅡcollagen,proteoglycan,Sox9 mRNA),and western blot assay(typeⅡcollagen,proteoglycan,Sox9 protein expression)were performed 12 weeks after surgery.RESULTS AND CONCLUSION:(1)X-ray films showed that the height of the intervertebral disc in the model group gradually decreased at 4,8,and 12 weeks after the operation,while the height of the intervertebral disc in the control group and the experimental group increased gradually.Among them,the rabbit disc height in the experimental group recovered better than the control group.(2)MRI demonstrated that at 4,8,and 12 weeks after surgery,the T2 weighted image signal value of the rabbit intervertebral disc in the model group gradua

关 键 词：材料 丝素蛋白 纳米羟基磷灰石 水凝胶 椎间盘退变 淫羊藿苷 实验动物 

分 类 号：R459.9[医药卫生—治疗学] R318.8[医药卫生—临床医学]