摘要
目的评估比格犬内皮祖细胞(EPCs)对组织工程骨的血管化和成骨效果。方法将比格犬骨髓来源的骨髓间充质干细胞(BMSCs)和EPCs进行分离、培养、扩增,在β-磷酸三钙(β-TCP)支架材料上进行接种。将EPCs/BMSCs/TCP、BMSCs/TCP细胞支架复合物和空白TCP分别植入比格犬下肢肌肉内,于术后3、6个月进行CT平扫,计算骨的相对CT值,并于术后6个月取材,行HE染色和免疫组织化学染色观察血管化和成骨效果。结果术后3个月,EPCs/BMSCs/TCP组的相对CT值为(366. 67±19. 51) HU,明显高于BMSCs/TCP组的(163. 00±30. 81) HU (t=2. 10,P=0. 0006);术后6个月, EPCs/BMSCs/TCP组的相对CT值为(553. 34±26. 86) HU,也明显高于BMSCs/TCP组的(241. 34±21. 57) HU (t=2. 11,P=0. 0006); BMSCs/TCP组(t=2. 10,P=0. 0255)和EPCs/BMSCs/TCP组(t=2. 10,P=0. 0006)术后6个月的相对CT值均明显高于该组术后3个月的相对CT值。术后6个月,HE染色显示空白TCP组未见骨结构形成和钙质沉积,EPCs/BMSCs/TCP组和BMSCs/TCP组可见成骨细胞、成熟的骨小梁及钙质沉积。免疫组织化学染色结果显示,EPCs/BMSCs/TCP组的外周、中心和整个支架的血管数量分别为(21. 67±1. 45)、(23. 33±2. 60)和(45. 00±1. 16)支,均明显高于BMSCs/TCP组的(8. 67±0. 88)(t=2. 07,P=0. 0016)、(9. 33±0. 67)(t=2. 07,P=0. 0065)和(18. 00±1. 00)支(t=2. 07,P=0. 001); BMSCs/TCP组与空白TCP组整个支架的血管数量差异无统计学意义[(18. 00±1. 00)支比(16. 67±2. 40)支; t=2. 07,P=0. 636]。EPCs/BMSCs/TCP组的成骨面积为(1 322 000±141300)像素,明显高于BMSCs/TCP组的(874 900±49 430)像素(t=2. 10,P=0. 04)。BMSCs/TCP组边缘区域的成骨面积明显高于中心区域[(170 000±42 320)像素比(613 900±90 290)像素; t=2. 10,P=0. 02]; EPCs/BMSCs/TCP组边缘区域和中心区域的成骨面积差异无统计学意义[(376 400±20 160)像素比(310 400±6917)像素; t=2. 10,P=0. 07]。结论 EPCs作为种子细胞联合BMSCs可在比格犬体内促进组织工程骨�
Objective The explore the effects of endothelial progenitor cells(EPCs)on vascularization and osteogenesis of tissue-engineered bones in Beagle dogs.Methods Bone marrow mesenchymal stem cells(BMSCs)and EPCs from bone marrow of Beagle dogs were isolated,cultured,and expanded and then seeded inβ-tricalcium phosphate(TCP)scaffolds.The cell scaffold complexes(EPCs/BMSCs/TCP group and BMSCs/TCP group)and the blank TCP scaffold were transplanted into the limb muscles of Beagle dogs,respectively.The relative CT values were calculated in the 3rd and 6th month through CT scan after operation.The vascularization and the osteogenesis were detected 6 months after operation by histological examination.Results Three months after the surgery,the relative CT value in the EPCs/BMSCs/TCP group was(366.67±19.51)HU,significantly higher than that[(163.00±30.81)HU]in the BMSCs/TCP group(t=2.10,P=0.0006);6 months after surgery,the relative CT value in the EPCs/BMSCs/TCP group was(553.34±26.86)HU,which was also significantly higher than that[(241.34±21.57)HU]in the BMSCs/TCP group(t=2.11,P=0.0006).The relative CT values at 6 months after operation were significantly higher than those at 3 months after operation in both BMSCs/TCP group(t=2.10,P=0.0255)and EPCs/BMSCs/TCP group(t=2.10,P=0.0006).Six months after the operation,HE staining showed no bone formation or calcium deposition in the blank TCP group,and osteoblasts,mature bone trabeculae,and calcium deposition were observed in EPCs/BMSCs/TCP group and BMSCs/TCP group.Immunohistochemical staining showed that the number of peripheral,central,and whole vessels in the EPCs/BMSCs/TCP group was 21.67±1.45,23.33±2.60,and 45.00±1.16,which were significantly higher than those in the BMSCs/TCP group[8.67±0.88(t=2.07,P=0.0016),9.33±0.67(t=2.07,P=0.0065),and 18.00±1.00(t=2.07,P=0.001),respectively].The number of vessels was not significantly different between the BMSCs/TCP group and the blank TCP group(18.00±1.00 vs.16.67±2.40;t=2.07,P=0.636).The osteogenic area of the EPCs/BMSCs/TC
作者
吴骁伟
尹琎
韦颍昕
WU Xiaowei;YIN Jin;WEI Yingxin(Department of Thoracic Surgery,Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology,Wuhan 430030,China;Department of Hematology,Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology,Wuhan 430030,China;Department of General Surgery,PUMC Hospital,CAMS and PUMC,Beijing 100730,China)
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2018年第5期642-650,共9页
Acta Academiae Medicinae Sinicae
基金
华中科技大学自主创新研究基金(2017KFYXJJ115)~~
关键词
组织工程骨
骨髓间充质干细胞
内皮祖细胞
tissue engineered bone
bone marrow mesenchymal stem cells
endothelial progenitor cells
