EPCs与BMSCs干细胞联合培养体系修复机体骨缺损

赵娴; 韩雪松; 周箐竹; 李杉; 刘流; 王松梅

EPCs与BMSCs干细胞联合培养体系修复机体骨缺损

1. 昆明医科大学第一附属医院整形外科
2. 云南大学生命科学学院
3. 昆明医科大学公共卫生学院

基金项目:

基金：国家自然科学基金资助项目 (81460296); 云南省应用基础研究面上基金资助项目 (2017FB121); 云南省卫生科技后备人才计划资助项目 (H-201631);

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出版历程
- 收稿日期: 2017-09-24

The Bone Defect Repairment in Vivo with Co-cultured EPCs and BMSCs

Funds:

基金：国家自然科学基金资助项目 (81460296); 云南省应用基础研究面上基金资助项目 (2017FB121); 云南省卫生科技后备人才计划资助项目 (H-201631);

摘要

摘要: 目的为解决整形外科骨缺损修复, 促进移植骨成活, 应用干细胞联合培养体系复合部分脱蛋白生物骨 (PDPBB) , 对机体胫骨缺损进行修复.方法新西兰大耳兔18只, 分别抽取骨髓液及外周血提取骨髓间充质干细胞 (BMSCs) 及外周血内皮祖细胞 (EPCs) , 构建联合培养体系后, 复合PDPBB构建组织工程骨;于兔胫骨制造1 cm长骨缺损, 将组织工程骨植入缺损区, 分别于术后14 d、28 d及2个月观察骨缺损修复的情况.结果 BMSCs组、联合培养细胞组和空白组各个时间点及组间的吸光度值比较, 差异均有统计学意义 (P<0.001) ;组织工程骨植入体内后骨胶原蛋白含量逐渐升高, 各分组差异有统计学意义 (P<0.01) ;复合联合干细胞体系的PDPBB修复骨缺损能力最强, 修复了胫骨缺损区的结构和功能.结论 EPCs与BMSCs联合干细胞体系复合PDPBB构建的组织工程骨是良好的骨缺损修复材料.
- 组织工程骨 /
- 内皮组细胞 /
- 骨髓间充质干细胞 /
- 联合培养 /
- 骨缺损修复
Abstract: Objective To reconstruct the deformity of appearance and function of patients with bone defect, co-cultured system with two stem cells were combined with partial deproteinized biological bone to reconstruct the defect of tibia which is one of the main weight-bearing bone. Methods The bone marrow and peripheral blood were harvested form 18 New Zealand rabbits to isolated bone marrow stem cells and epithelial progenitor cells, and engineering bone was constructed with co-cultured system with these two stem cells and partial deproteinized biological bones; about 1 CM of bone defect of each rabbit was made with bone rongeur, then engineering bones were transplanted into the defect area, the osteogenesis and bone defect recovery were observed on day 14, 28 and month 2.Results The difference of absorbance values of BMSCs group, co-cultured cell group and blank group at each time point and between groups were all statistically significant (P<0.001) , and the collagen content of bone tissue increased gradually after implantation of tissue engineered bone, and the difference between each group was statistically significant (P <0.001) . The repairment of bone defect with the PDPBB combined with BMSCs and EPCs system has the strongest ability to repair the structure andfunction of the tibial defect area. Conclusion The engineering bone constructed with two stem cells and partial deproteinized bone is a good material for bone defect reconstruction.
- Engineering bone /
- Epithelial progenitor cell /
- Bone marrow mesenchymal stem cell /
- Co-culture /
- Bone defect repairment

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参考文献(19)

[1]	[1]KANCZLER J M, OREFFO R.Osteogenesis and angiogenesis:The potential for engineering bone[J].European Cells and Material, 2008, 15 (1) :100-114.
[2]	[2]MUELLER S, GOSAU M, WENDY C M, et al.Postoperative evaluation of microvascularization in mandibular 290 reconstructions with microvascular flaps-first results with a new perfusion software for contrast-enhanced sonography (CEUS) [J].Clin Hemorheol Microcirc, 2012, 52 (2-4) :187-196.
[3]	[3]COLNOT C, ROMERO D M, HUANG S, et al.Mechanisms of action of demineralized bone matrix in the repair of cortical bone defects[J].Clin Orthop Relat Res, 2005, 435 (435) :69-78.
[4]	[4]BRANDI ML, COLLIN-OSDOBY P.Vascular biology and the skeleton[J].J Bone Miner Res, 2006, 21 (2) :183-192.
[5]	[5]COULTAS L, CHAWENGSAKSOPHAK K, ROSSANT J.Endothelial cells and VEGF in vascular development[J].Nature, 2005, 438 (7070) :937-945.
[6]	[6]JAIN R K.Molecular regulation of vessel maturation[J].Nat Med, 2003, 6 (6) :685-693.
[7]	[7]BILOUSOVA G, JUN DU H, KING KB, et al.Osteoblasts derived from induced pluripotent stem cells form calcified structrues in scaffolds both in vitro and in vivo[J].Stem Cells, 2011, 29 (2) :206-216.
[8]	[8]WHEELER D L, LANE J M, SEIM H B, et al.Allogenetic mesenchymal progenitor cells for posterolateral lumbar spine fusion in sheep[J].Spine J, 2014, 14 (3) :435-444.
[9]	[9]LARGER M, PRISBY R, PETER Z, et al.Quantitative investigation of bone microvascularization from 3D synchrotron micro-computed tomography in a rat model[J].Conf Proc IEEE Eng Med Biol Soc, 2009, 1 (1) :1004-1007.
[10]	[10]WERNIKE E, MONTJOVENT M O, LIU Y, et al.VEGF incorporated into calciuma phosphated ceramics promotes 305 vascularisation and bone formation in vivo[J].Eur Cell Mater, 2010, 22 (19) :30-40.
[11]肖仕辉, 韦积华, 韦庆军.组织工程血管化的研究进展[J].广东医学, 2012, 33 (11) :1680-1682.
[12]	[12]YAMADA Y, UEDA M, HIBI H, et al.Translational research for inject able tissue-engineered bone regeneration using mesenchymal stem cells and platelet-rich plasma:from basic research to clinical case study[J].Cell Transplant, 2004, 13 (4) :343-355.
[13]	[13]CHOONG C S, HUTMACHER D W, TRIFFITT J T.Co-culture of bone marrow fibroblasts and endothelial cells on modified polycaprolactone substrates for enhanced potentials in bone tissue engineering[J].Tissue Eng, 2006, 12 (9) :2521-2531.
[14]	[14]DECKERS M M, VAN BEZOOIJEN R L, VAN DER HORST G, et al.Bone morphogenetic proteins stimulate angiogenesis through osteroblast-derived vascular endothelial growth factor A[J].Endocrinology, 2002, 143 (4) :1545-1553.
[15]	[15]MUROTA S I, ONODERA M, MORITA I.Regulation of angiogenesis by controlling VEGF receptor[J].Ann N Y Acad Sci, 2000, 902 (1) :208-212.
[16]	[16]PALMIERI D, CAMARDELLA L, ULIVI V, et al.Trimer carboxyl propeptide of collagen I produced by mature osteoblasts ischemotactic for endothelial cells[J].J Biol Chem, 2000, 275 (42) :32658-32663.
[17]	[17]VELAZQUEZ O C, SNYDER R, LIU Z J, et al.Fibroblast-dependent differentiation of human microvascular320 endothelial cells into capillary-like 3-dimensional networks[J].FASEB J, 2002, 16 (10) :1316-1318.
[18]	[18]PHILIPPE P, FRANK P, SIMONE M P, et al.Influences of visualization and osteogenic cells on heterotopic bone formation within a madreporci ceramic in rats[J].Plast Surg, 2003, 111 (6) :1932-1941.
[19]	[19]VILLARS F, BORDENAVE L, BAREILLE R, et al.Effect of human endothelial cells on human bone marrow stromal cell phenotype:role of VEGF[J].J Cell Biochem, 2000, 79 (4) :672-685.

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