摘要
背景:股骨头坏死形成机制中,脂代谢紊乱学说受到许多学者的重视。茶叶提取物茶黄素具有降脂作用,在骨髓间充质干细胞培养中可抑制激素诱导的成脂分化,加强其成骨能力。目的:观察骨髓间充质干细胞联合茶黄素治疗激素性股骨头坏死的效果。设计、时间及地点:随机对照动物实验,于2008-03/06在安徽医科大学实验中心完成。材料:洁净级健康成年新西兰大白兔24只,随机分为4组:模型对照组、单纯减压组、细胞移植组、细胞联合茶黄素组,6只/组。实验所用茶黄素为茶黄素单体,纯度>60%,由上海嘉会生物科技有限公司生产,批号20040828。方法:体外分离培养兔自体骨髓间充质干细胞,传至第3代后调整细胞密度为2×1010L-1复合于3cm×2cm明胶海绵上,每块明胶海绵含细胞悬液60μL,在细胞培养箱中孵育4h备用。各组兔均采用液氮冷冻法建立激素性股骨头坏死模型,自然复温后钻孔,模型对照组不作任何处理,单纯减压组植入明胶海绵,细胞移植组、细胞联合茶黄素组植入复合有兔自体骨髓间充质干细胞的明胶海绵,同时细胞联合茶黄素组每天分3次灌服茶黄素250mg,直至处死。主要观察指标:动物一般情况,髋部MRI扫描观察股骨头坏死区信号变化,以及股骨头标本组织学观察、扫描电镜观察结果。结果:造模后8周,细胞移植组、细胞联合茶黄素组兔活动明显增加,走路姿势逐步好转。造模后4周,各组均未见明显坏死区信号改变;造模后8周,模型对照组坏死低信号区明显扩大,单纯减压组低信号区无明显变化,细胞移植组低信号区范围略微缩小,细胞联合茶黄素组低信号区明显缩小,骨密度信号接近正常。与其余3组比较,造模后4,8周细胞联合茶黄素组空骨陷窝阳性数均明显减少(P<0.05)。扫描电镜下,造模后8周模型对照组骨小梁多处断裂塌陷,髓腔内有大量脂肪细胞堆积;单纯减压组部分骨小梁可见裂痕;细胞移植组骨小梁不致密;细胞联合茶黄素组骨小梁结构规整致密,成骨细胞多,骨细胞及基质胶原纤维正常,髓腔规则。结论:茶黄素能促进骨髓间充质干细胞成骨,在髓芯减压并骨髓间充质干细胞移植的基础上灌服给予茶黄素治疗兔激素性股骨头坏死的效果较好。
BACKGROUND： Of the mechanisms of femoral head necrosis formation, lipid metabolic disorder has received much attention by many scholars. Theaflavin extracted from tea can decrease lipid, inhibit hormone-derived adipocytic differentiation, and strengthen osteogenic ability during bone marrow mesenchymal stem cell （BMSC） culture, OBJECTIVE： To observe curative effects of combination of theaflavin and BMSCs for treating steroid-associated osteonecrosis of femoral head. DESIGN, TIME AND SETTING： The randomized controlled animal study was performed at the Experimental Center of Anhui Medical University from March to June 2008. MATERIALS： A total of 24 clean healthy adult New Zealand rabbits were equally and randomly divided into model control, decompression, BMSC transplantation, theaflavin ＋ BMSC transplantation groups. Theaflavin used in this study was theaflavin monomer, with a pudty of 〉60% （lot number 20040828, Jiahui, Shanghai, China）. METHODS： Rabbit autologous BMSCs were isolated and cultured in vitro. At the third passage, BMSCs at 2×10^10/L were combined on 3 cm×2 cm gelatin sponge. Each gelatin sponge contained 60 μL cell suspension, placed in a cell incubator for 4 hours for further use. in each group, rabbit models of steroid-associated femoral head necrosis were established by liquid nitrogen frozen method. After natural rewarming, a hole was drilled. Rabbits in the model control group were left intact. Rabbits in the decompression group were implanted with gelatin sponge. Rabbits of the BMSC transplantation and theaflavin ＋ BMSC transplantation groups were implanted with gelatin sponge with rabbit autologous BMSCs, and perfused with 250 mg theaflavin three time every day till death. MAIN OUTCOME MEASURES： The following parameters were measured： general conditions of animals; signal changes in the femoral head necrosis region were observed in the hip by magnetic resonance imaging scanning. Histology of femoral head samples was observed. Results of scanning electron microscope observation. RESULTS： After 8 weeks, rabbits became more active than before treatment, and walking way became normal gradually in the BMSC transplantation, theaflavin ＋ BMSC transplantation groups. Four weeks after model induction, magnetic resonance imaging signal region of all groups had no obvious changes, but 8 weeks later, the necrosis signal region of the model control group magnified while it reduced obviously in the decompression group. Low signal region range became slightly small in the cell transplantation group. The low signal region range became significantly small in the theaflavin ＋ BMSC transplantation group, and bone density signal was nearly normal. Compared with other three groups, positive number of empty lacunae significantly reduced in the theaflavin ＋ BMSC transplantation group at 4 and 8 weeks following model induction （P 〈 0.05）. Under the scanning electron microscope, after 8 weeks of treatment, the bone trabecula collapsed in many regions, and a large number of adipocytes accumulated in the medullary ccavity in the model control group. Fracture of some bone trabecula appeared in the decompression group. Bone trabecula was not dense in the BMSC transplantation group. Bone trabecula was closely dense, with many osteoblasts, normal osteocytes and stromal collagen fiber and regular medullary cavity in the theaflavin ＋ BMSC transplantation group. CONCLUSION： Theaflavin can promote the differentiation of osteocyte from BMSCs. The combination of theaflavin and BMSC transplantation based on core decompression for the treatment of steroid-associated osteonecrosis of femoral head have good application prospects.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第19期3730-3734,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
安徽省自然科学基金资助项目(070413083)~~
