VEGF转染MSCs结合PLA/HA多孔支架治疗兔早期股骨头坏死的实验研究
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摘要
第一部分激素联合脂多糖诱导早期兔股骨头坏死的实验研究
目的运用组织学技术、CT扫描和显微CT成像观察激素联合脂多糖诱导早期兔股骨头坏死的实验效果。
方法健康雄性新西兰大白兔30只,随机分为实验组25只和对照组5只,实验组经耳缘静脉注射10μg/Kg的脂多糖二次,再肌肉注射20 mg/Kg甲泼尼龙三次,对照组注射同等剂量的生理盐水,5周后CT扫描髋关节,随机处死实验组和对照组兔4只,显微CT成像扫描股骨近端,并作组织切片观察。结果CT扫描显示实验组兔股骨头不同程度的骨密度不均匀,显微CT成像显示骨质疏松,软骨下骨囊性化,周围见硬化骨,组织学切片见骨细胞陷窝空疏,脂肪细胞增多,部分血管栓塞。
结论激素联合脂多糖能够安全便捷地诱导兔早期股骨头坏死模型。
第二部分VEGF基因转染MSCs结合PLA/HA复合多孔支架的异位成骨研究
目的观察pcDNA3.1-VEGF165重组质粒转染大鼠骨髓间充质干细胞(MSCs)结合聚乳酸/纳米羟基磷灰石复合多孔支架的异位成骨效果。方法实验分为3组：(1)VEGF转染MSCs复合材料组,(2)空白载体转染MSCs复合材料组,(3)单纯支架组。分离大鼠骨髓间充质干细胞,诱导成骨条件培养,基因转染后RT-PCR和Western blot方法检测VEGF的表达,细胞与材料复合后扫描电镜观察。将复合细胞的支架材料植入大鼠肌袋后第2,4,8周进行X线和组织学检测。结果成功分离出大鼠骨髓基质干细胞,并诱导成骨方向培养,RT-PCR和Western blot显示转染后MSCs的VEGF表达水平明显高于其他2组,聚乳酸/纳米羟基磷灰石复合多孔支架呈三维多孔隙结构,细胞能有效生长,植入大鼠体内后,转染组4周可见异位成骨,8周更加明显,另2组未见明显成骨。
结论VEGF基因转染大鼠MSCs复合PLA/HA多孔支架能在大鼠体内诱导成骨。
第三部分VEGF基因转染MSCs结合PLA/HA复合多孔支架治疗兔早期股骨头坏死
目的探讨利用pcDNA3.1-VEGF165重组质粒转染兔MSCs与PLA/HA多孔支架复合后植入股骨头坏死模型兔的股骨头内,观察其成血管和成骨能力。
方法分离、培养兔骨髓基质干细胞,pcDNA3.1-VEGF165重组质粒转染兔MSCs后RT-PCR检测VEGF表达情况,将转染后的兔MSCs与PLA/HA多孔支架复合。实验分为四组：(1)pcDNA3.1-VEGF165重组质粒转染MSCs复合支架组,(2)空白载体转染MSCs复合支架组,(3)单纯支架植入组,(4)未处理组。经兔股骨大转子后下方向股骨头钻直径3mm工作隧道,植入支架材料。术后分别于8、12周对股骨头行显微CT及组织病理学切片检查,第12周行X线摄影。
结果成功分离培养出兔MSCs, pcDNA3.1-VEGF165重组质粒转染后,VEGF表达水平明显高于其他组。pcDNA3.1-VEGF165重组质粒转染MSCs支架植入股骨头后8周可见坏死骨小梁周围部分新骨形成,支架周围成纤维细胞、淋巴细胞浸润,小血管形成,12周见新生骨小梁生成,支架吸收,而其他组未见明显新生骨。显微CT显示重组质粒转染组股骨头形态正常,骨小梁分布均匀,其他组部分股骨头变形,软骨下骨变薄,骨小梁稀疏、断裂。X线显示重组质粒转染组股骨头形态正常、密度均匀,未治疗组股骨头明显变形,骨密度不均匀。
结论pcDNA3.1-VEGF165重组质粒转染兔MSCs复合PLA/HA多孔支架能够诱导兔股骨头内新生血管和骨的形成,是一种较好的修复股骨头坏死的方法。
The experimental research of early osteonecrosis of rabbit femoral head induced by a comibination of methylprednisolone and lipopolysaccharide
objective To research the experimental result of early osteonecrosis of rabbit femoral head induced by a comibination of methylprednisolone and lipopolysaccharide by histopathologic technique, CT scanning and micro CT imaging.
Methods 30 male new-Zealand rabbits were divided into experimental group (25 rabbits) and control group (5 rabbits) randomly. Two injections of 10μg/Kg body weight of lipopolysaccharide and three injections of 20 mg/Kg body weight of methylprednisolone were performed in experimental group. Physiologic saline was injected in control group. We scanned the hip joint of every rabbit by CT, and executed 4 rabbits in both groups randomly for micro CT sacnning and histopathology.
Results The CT scanning showed non-uniform of bone density of femoral head in experimental group. Osteoporosis, cystis and sclerotization were observed in subchondral bone from micro CT imaging. Bone cell lacunae, increased fat cells and thrombokinesis were observed in histopathologic check.
Conclusion A comibination of methylprednisolone and lipopolysaccharide could induce the early osteonecrosis of rabbit femoral head safely.
objective To evaluate the ectopic osteogenesis potential of polylactic acid/hydroxyapatite ceramic (PLA/HA) porous scaffold delivery of rat bone marrow-derived mesenchymal stem cells transfected by pcDNA3.1-VEGF165 plasmid.
Methods Rat MSCs were separated and cultured with osteoinduction medium. Cells were transfected with pcDNA3.1-VEGF165 plasmid or blank plasmid. RT-PCR and Western blot were used to determine the phenotype of MSCs. There were three groups:plasmid transfection with scaffold group, blank plasmid transfection with scaffold group and only scaffold group. Scanning electron microscope was used to analyzed the biocompatibility of scaffold. Cells-scaffold complexes were implanted into the muscles of rat. Radiographic and histological evaluations were performed to observed the bone formation 2,4 and 8 weeks after operation.
Results Rat MSCs were sucessed to be separated and cultured with osteoinduction medium. By RT-PCR test and Western blot analysis, only the pcDNA3.1-VEGF165 transfected MSCs producted VEGF165. Scanning electron microscope showed the 3D-structure with more pores of PLA/HA porous scaffold.4 weeks after operation, ectopic bones were observed in plasmid transfection with scaffold group. little ectopic bones were observed in other groups.
Conclusions PLA/HA porous scaffold delivery of rat MSCs transfected by pcDNA3.1-VEGF165 plasmid can induce ectopic osteogenesis of rat in vivo.
Objective To observe the blood vessels and osteogenic capacity after implantation the combination of plasmid pcDNA3.1-VEGF165 transfection of rabbit bMSCs with the PLA/HA porous scaffold into the femoral head of rabbit femoral head necrosis model.
Methods Rabbit MSCs were isolated and cultured. After transfected with pcDNA3.1-VEGF165 plasmid, we detected VEGF expression by RT-PCR, then we combinated rabbit MSCs with the PLA/HA porous scaffold. There were 4 groups:(1) pcDNA3.1-VEGF165 plasmid transfection with scaffold group, (2)blank vector transfection with scaffold group, (3)simple scaffold group,(4)untreated group. We drilled 3mm diameter tunnel under the greater trochanter to femoral head, and implanted porous scaffolds. Micro-CT and histological evaluations were performed to observe the rabbit femoral head 8 and 12 weeks after operation. X-ray photography was performed after 12 weeks.
Result Rabbit MSCs were separated and cultured with osteoinduction medium successfully. After transfected with recombinant plasmid pcDNA3.1-VEGF165, VEGF expressipn level was significantly higher than other groups.8 weeks after operation, we can observe the new bone formation around the necrotic trabecular, ibroblast cells and lymphocyte infiltration, small blood vessel formation in recombinant plasmid transfected MSC group. After 12 weeks, we saw the new trabecular bone generation and scaffold absorption, while the other group had no obvious new bone. Micro-CT showed normal femoral head shape, uniform distribution of trabecular bone in plasmid transfection group, but femoral head deformation, thin subchondral bone and trabecular bone fracture in other groups. X-ray showed morphologically normal femoral heads and uniform bone mineral density in plasmid transfection group, but femoral head deformation obviously and non-uniform bone mineral density in untreated group.
Conclusion Plasmid pcDNA3.1-VEGF165 transfection of rabbit MSCs with the PLA/HA porous scaffold could induce the formation of blood vessels and bone in rabbit femoral head, which is a better method for repair of femoral head necrosis.
目的运用组织学技术、CT扫描和显微CT成像观察激素联合脂多糖诱导早期兔股骨头坏死的实验效果。
方法健康雄性新西兰大白兔30只,随机分为实验组25只和对照组5只,实验组经耳缘静脉注射10μg/Kg的脂多糖二次,再肌肉注射20 mg/Kg甲泼尼龙三次,对照组注射同等剂量的生理盐水,5周后CT扫描髋关节,随机处死实验组和对照组兔4只,显微CT成像扫描股骨近端,并作组织切片观察。结果CT扫描显示实验组兔股骨头不同程度的骨密度不均匀,显微CT成像显示骨质疏松,软骨下骨囊性化,周围见硬化骨,组织学切片见骨细胞陷窝空疏,脂肪细胞增多,部分血管栓塞。
结论激素联合脂多糖能够安全便捷地诱导兔早期股骨头坏死模型。
第二部分VEGF基因转染MSCs结合PLA/HA复合多孔支架的异位成骨研究
目的观察pcDNA3.1-VEGF165重组质粒转染大鼠骨髓间充质干细胞(MSCs)结合聚乳酸/纳米羟基磷灰石复合多孔支架的异位成骨效果。方法实验分为3组：(1)VEGF转染MSCs复合材料组,(2)空白载体转染MSCs复合材料组,(3)单纯支架组。分离大鼠骨髓间充质干细胞,诱导成骨条件培养,基因转染后RT-PCR和Western blot方法检测VEGF的表达,细胞与材料复合后扫描电镜观察。将复合细胞的支架材料植入大鼠肌袋后第2,4,8周进行X线和组织学检测。结果成功分离出大鼠骨髓基质干细胞,并诱导成骨方向培养,RT-PCR和Western blot显示转染后MSCs的VEGF表达水平明显高于其他2组,聚乳酸/纳米羟基磷灰石复合多孔支架呈三维多孔隙结构,细胞能有效生长,植入大鼠体内后,转染组4周可见异位成骨,8周更加明显,另2组未见明显成骨。
结论VEGF基因转染大鼠MSCs复合PLA/HA多孔支架能在大鼠体内诱导成骨。
第三部分VEGF基因转染MSCs结合PLA/HA复合多孔支架治疗兔早期股骨头坏死
目的探讨利用pcDNA3.1-VEGF165重组质粒转染兔MSCs与PLA/HA多孔支架复合后植入股骨头坏死模型兔的股骨头内,观察其成血管和成骨能力。
方法分离、培养兔骨髓基质干细胞,pcDNA3.1-VEGF165重组质粒转染兔MSCs后RT-PCR检测VEGF表达情况,将转染后的兔MSCs与PLA/HA多孔支架复合。实验分为四组：(1)pcDNA3.1-VEGF165重组质粒转染MSCs复合支架组,(2)空白载体转染MSCs复合支架组,(3)单纯支架植入组,(4)未处理组。经兔股骨大转子后下方向股骨头钻直径3mm工作隧道,植入支架材料。术后分别于8、12周对股骨头行显微CT及组织病理学切片检查,第12周行X线摄影。
结果成功分离培养出兔MSCs, pcDNA3.1-VEGF165重组质粒转染后,VEGF表达水平明显高于其他组。pcDNA3.1-VEGF165重组质粒转染MSCs支架植入股骨头后8周可见坏死骨小梁周围部分新骨形成,支架周围成纤维细胞、淋巴细胞浸润,小血管形成,12周见新生骨小梁生成,支架吸收,而其他组未见明显新生骨。显微CT显示重组质粒转染组股骨头形态正常,骨小梁分布均匀,其他组部分股骨头变形,软骨下骨变薄,骨小梁稀疏、断裂。X线显示重组质粒转染组股骨头形态正常、密度均匀,未治疗组股骨头明显变形,骨密度不均匀。
结论pcDNA3.1-VEGF165重组质粒转染兔MSCs复合PLA/HA多孔支架能够诱导兔股骨头内新生血管和骨的形成,是一种较好的修复股骨头坏死的方法。
The experimental research of early osteonecrosis of rabbit femoral head induced by a comibination of methylprednisolone and lipopolysaccharide
objective To research the experimental result of early osteonecrosis of rabbit femoral head induced by a comibination of methylprednisolone and lipopolysaccharide by histopathologic technique, CT scanning and micro CT imaging.
Methods 30 male new-Zealand rabbits were divided into experimental group (25 rabbits) and control group (5 rabbits) randomly. Two injections of 10μg/Kg body weight of lipopolysaccharide and three injections of 20 mg/Kg body weight of methylprednisolone were performed in experimental group. Physiologic saline was injected in control group. We scanned the hip joint of every rabbit by CT, and executed 4 rabbits in both groups randomly for micro CT sacnning and histopathology.
Results The CT scanning showed non-uniform of bone density of femoral head in experimental group. Osteoporosis, cystis and sclerotization were observed in subchondral bone from micro CT imaging. Bone cell lacunae, increased fat cells and thrombokinesis were observed in histopathologic check.
Conclusion A comibination of methylprednisolone and lipopolysaccharide could induce the early osteonecrosis of rabbit femoral head safely.
objective To evaluate the ectopic osteogenesis potential of polylactic acid/hydroxyapatite ceramic (PLA/HA) porous scaffold delivery of rat bone marrow-derived mesenchymal stem cells transfected by pcDNA3.1-VEGF165 plasmid.
Methods Rat MSCs were separated and cultured with osteoinduction medium. Cells were transfected with pcDNA3.1-VEGF165 plasmid or blank plasmid. RT-PCR and Western blot were used to determine the phenotype of MSCs. There were three groups:plasmid transfection with scaffold group, blank plasmid transfection with scaffold group and only scaffold group. Scanning electron microscope was used to analyzed the biocompatibility of scaffold. Cells-scaffold complexes were implanted into the muscles of rat. Radiographic and histological evaluations were performed to observed the bone formation 2,4 and 8 weeks after operation.
Results Rat MSCs were sucessed to be separated and cultured with osteoinduction medium. By RT-PCR test and Western blot analysis, only the pcDNA3.1-VEGF165 transfected MSCs producted VEGF165. Scanning electron microscope showed the 3D-structure with more pores of PLA/HA porous scaffold.4 weeks after operation, ectopic bones were observed in plasmid transfection with scaffold group. little ectopic bones were observed in other groups.
Conclusions PLA/HA porous scaffold delivery of rat MSCs transfected by pcDNA3.1-VEGF165 plasmid can induce ectopic osteogenesis of rat in vivo.
Objective To observe the blood vessels and osteogenic capacity after implantation the combination of plasmid pcDNA3.1-VEGF165 transfection of rabbit bMSCs with the PLA/HA porous scaffold into the femoral head of rabbit femoral head necrosis model.
Methods Rabbit MSCs were isolated and cultured. After transfected with pcDNA3.1-VEGF165 plasmid, we detected VEGF expression by RT-PCR, then we combinated rabbit MSCs with the PLA/HA porous scaffold. There were 4 groups:(1) pcDNA3.1-VEGF165 plasmid transfection with scaffold group, (2)blank vector transfection with scaffold group, (3)simple scaffold group,(4)untreated group. We drilled 3mm diameter tunnel under the greater trochanter to femoral head, and implanted porous scaffolds. Micro-CT and histological evaluations were performed to observe the rabbit femoral head 8 and 12 weeks after operation. X-ray photography was performed after 12 weeks.
Result Rabbit MSCs were separated and cultured with osteoinduction medium successfully. After transfected with recombinant plasmid pcDNA3.1-VEGF165, VEGF expressipn level was significantly higher than other groups.8 weeks after operation, we can observe the new bone formation around the necrotic trabecular, ibroblast cells and lymphocyte infiltration, small blood vessel formation in recombinant plasmid transfected MSC group. After 12 weeks, we saw the new trabecular bone generation and scaffold absorption, while the other group had no obvious new bone. Micro-CT showed normal femoral head shape, uniform distribution of trabecular bone in plasmid transfection group, but femoral head deformation, thin subchondral bone and trabecular bone fracture in other groups. X-ray showed morphologically normal femoral heads and uniform bone mineral density in plasmid transfection group, but femoral head deformation obviously and non-uniform bone mineral density in untreated group.
Conclusion Plasmid pcDNA3.1-VEGF165 transfection of rabbit MSCs with the PLA/HA porous scaffold could induce the formation of blood vessels and bone in rabbit femoral head, which is a better method for repair of femoral head necrosis.
引文
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18.方园 戴红莲 李世普.液相吸附法制备羟基磷灰石/聚乳酸复合生物材料.生物骨科材料与临床研究,2006；3(4)：38-41.
19. Rodrigues CV, Serricella P, Linhares AB, et al. Characterization of a bovine collagen-hydroxyapatite composite scaffold for bone tissue engineering. Biomaterials, 2003; 24(27):4987-4997.
20.郭晓东,郑启新,杜靖远等.可吸收羟基磷灰石/聚DL-乳酸骨折内固定材料机械强度和生物降解性研究.中国生物医学工程学报,2001；20(1)：23-28.
21. Maes C, Carmeliet P, Moermans K, et al. Impaired angiogenesis and endochondral bone formation in mice lacking the vascular endothelial growth factor isoforms VEGF164 and VEGF 188. Mech Dev,2002; 111(1-2):61-73.
22. Mayer H, Bertram H, Lindenmaier W, et al. Vascular endothelial growth factor (VEGF-A) expression in human mesenchymal stem cells:autocrine and paracrine role on osteoblastic and endothelial differentiation. J Cell Biochem,2005; 95(4):827-839.
23. Deckers MM, van Bezooijen RL, van der Horst G, et al. Bone morphogenetic proteins stimulate angiogenesis through osteoblast-derived vascular endothelial growth factor A. Endocrinology,2002,143(4):1545-1553.
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2. Steinberg ME, Larcom PG, Strafford B, et al. Core decompression with bone grafting for osteonecrosis of the femoral head. Clin Orthop Relat Res,2001; (386):71-78.
3. Hernigou P, Beaujean F, Lambotte JC. Decrease in the mesenchymal stem-cell pool in the proximal femur in corticosteroid-induced osteonecrosis. J Bone Joint Surg Br, 1999; 81(2):349-355.
4. Suh KT, Kim SW, Roh HL, et al. Decreased osteogenic differentiation of mesenchymal stem cells in alcohol-induced osteonecrosis. Clin Orthop Relat Res,2005; (431): 220-225.
5. Lee JS, Lee JS, Roh HL, et al. Alterations in the differentiation ability of mesenchymal stem cells in patients with nontraumatic osteonecrosis of the femoral head:comparative analysis according to the risk factor. J Orthop Res,2006; 24(4):604-609.
6. Lee HS, Huang GT, Chiang H, et al. Multipotential mesenchymal stem cells from femoral bone marrow near the site of osteonecrosis. Stem Cells,2003; 21(2):190-199.
7. Deten A, Volz HC, Clamors S, et al. Hematopoietic stem cells do not repair the infarcted mouse heart. Cardiovasc Res,2005; 65(1):52-63.
8. Priller J. Robert Feulgen Prize Lecture. Grenzganger:adult bone marrow cells populate the brain. Histochem Cell Biol,2003; 120(2):85-91.
9. Wang Z, Goh J, Das De S, et al. Efficacy of bone marrow-derived stem cells in strengthening osteoporotic bone in a rabbit model. Tissue eng,2006; 12(7):1753-1761.
10. Gangji V, Hauzeur JP, Schoutens A, et al. Abnormalities in the replicative capacity of Osteoblastic cells in the proximal femur of patients with osteonecrosis of the femoral head. J Rheumatol,2003; 30(2):348-351.
11. Hernigou P, Beaujean F, Lambotte JC. Decrease in the mesenchymal stem-cell pool in the proximal femur in corticosteroid-induced osteonecrosis. Bone Joint Surg Br,1999; 81(2):349-355.
12. Tang TT, Lu B, Yue B, et al. Treatment of osteonecrosis of the femoral head with hBMP-2-gene-modified tissue-engineered bone in goats. J Bone Joint Surg Br,2007; 89(1):127-129.
13. Geiger F, Bertram H, Berger I, et al. Vascular endothelial growth factor gene-activated matrix (VEGF165-GAM) enhances osteogenesis and angiogenesis in large segmental bone defects. J Bone Miner Res.2005; 20(11):2028-2035.
14.Mayer H, Bertram H, Lindenmaier W, et al. Vascular endothelial growth factor (VEGF-A) expression in human mesenchymal stem cells:autocrine and paracrine role on osteoblastic and endothelial differentiation. J Cell Biochem,2005; 95(4):827-839.
15.Hofbaner LC, Heufclder AE. Updating the mctalopmtease nomenclature:bone morphogenetic protein 1 identified as procollagen C proteinase. Eur J Endocrinol, 1996; 135(1):35-36.
16.曹凯,黄伟,安洪等.VEGF基因转染结合脱蛋白骨修复股骨头坏死的实验研究.重庆医科大学学报,2005；30(4)：505-509.
17. Hiltunen MO, Ruuskanen M, Huuskonen J, et al.Adenovirus-mediated VEGF-A gene transfer induces bone formation in vivo. FASEB J,2003; 17(9):1147-1149.
18. Shuler MS, Rooks MD, Roberson JR. Porous tantalum implant in early osteonecrosis of the hip:preliminary report on operative, survival, and outcomes results. J Arthroplasty,2007; 22(1):26-31.
19. Stolzing A, Scutt A. Effect of reduced culture temperature on antioxidant defences of mesenchymal stem cells. Free Radic Biol Med,2006; 41(2):326-338.
20. Gangji V, Toungouz M, Hauzeur JP. Stem cell therapy for osteonecrosis of the femoral head. Expert Opin Biol Ther, 2005;5(4):437-442.
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1. Koike N, Fukumura D, Gralla O, et al. Tissue engineering:creation of long-lasting blood vessels. Nature 2004; 428:138-139.
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1. Babis GC, Soucacos PN. Efectiveness of total hip arthroplasty in the management of hip osteonecrosis. Orthop Clin North Am,2004,35(3):359-364.
2. Steinberg ME, Larcom PG, Strafford B, et al. Core decompression with bone grafting for osteonecrosis of the femoral head. Clin Orthop Relat Res,2001; (386):71-78.
3. Hernigou P, Beaujean F, Lambotte JC. Decrease in the mesenchymal stem-cell pool in the proximal femur in corticosteroid-induced osteonecrosis. J Bone Joint Surg Br, 1999; 81(2):349-355.
4. Suh KT, Kim SW, Roh HL, et al. Decreased osteogenic differentiation of mesenchymal stem cells in alcohol-induced osteonecrosis. Clin Orthop Relat Res,2005; (431): 220-225.
5. Lee JS, Lee JS, Roh HL, et al. Alterations in the differentiation ability of mesenchymal stem cells in patients with nontraumatic osteonecrosis of the femoral head:comparative analysis according to the risk factor. J Orthop Res,2006; 24(4):604-609.
6. Lee HS, Huang GT, Chiang H, et al. Multipotential mesenchymal stem cells from femoral bone marrow near the site of osteonecrosis. Stem Cells,2003; 21(2):190-199.
7. Deten A, Volz HC, Clamors S, et al. Hematopoietic stem cells do not repair the infarcted mouse heart. Cardiovasc Res,2005; 65(1):52-63.
8. Priller J. Robert Feulgen Prize Lecture. Grenzganger:adult bone marrow cells populate the brain. Histochem Cell Biol,2003; 120(2):85-91.
9. Wang Z, Goh J, Das De S, et al. Efficacy of bone marrow-derived stem cells in strengthening osteoporotic bone in a rabbit model. Tissue eng,2006; 12(7):1753-1761.
10. Gangji V, Hauzeur JP, Schoutens A, et al. Abnormalities in the replicative capacity of Osteoblastic cells in the proximal femur of patients with osteonecrosis of the femoral head. J Rheumatol,2003; 30(2):348-351.
11. Hernigou P, Beaujean F, Lambotte JC. Decrease in the mesenchymal stem-cell pool in the proximal femur in corticosteroid-induced osteonecrosis. Bone Joint Surg Br,1999; 81(2):349-355.
12. Tang TT, Lu B, Yue B, et al. Treatment of osteonecrosis of the femoral head with hBMP-2-gene-modified tissue-engineered bone in goats. J Bone Joint Surg Br,2007; 89(1):127-129.
13. Geiger F, Bertram H, Berger I, et al. Vascular endothelial growth factor gene-activated matrix (VEGF165-GAM) enhances osteogenesis and angiogenesis in large segmental bone defects. J Bone Miner Res.2005; 20(11):2028-2035.
14.Mayer H, Bertram H, Lindenmaier W, et al. Vascular endothelial growth factor (VEGF-A) expression in human mesenchymal stem cells:autocrine and paracrine role on osteoblastic and endothelial differentiation. J Cell Biochem,2005; 95(4):827-839.
15.Hofbaner LC, Heufclder AE. Updating the mctalopmtease nomenclature:bone morphogenetic protein 1 identified as procollagen C proteinase. Eur J Endocrinol, 1996; 135(1):35-36.
16.曹凯,黄伟,安洪等.VEGF基因转染结合脱蛋白骨修复股骨头坏死的实验研究.重庆医科大学学报,2005；30(4)：505-509.
17. Hiltunen MO, Ruuskanen M, Huuskonen J, et al.Adenovirus-mediated VEGF-A gene transfer induces bone formation in vivo. FASEB J,2003; 17(9):1147-1149.
18. Shuler MS, Rooks MD, Roberson JR. Porous tantalum implant in early osteonecrosis of the hip:preliminary report on operative, survival, and outcomes results. J Arthroplasty,2007; 22(1):26-31.
19. Stolzing A, Scutt A. Effect of reduced culture temperature on antioxidant defences of mesenchymal stem cells. Free Radic Biol Med,2006; 41(2):326-338.
20. Gangji V, Toungouz M, Hauzeur JP. Stem cell therapy for osteonecrosis of the femoral head. Expert Opin Biol Ther, 2005;5(4):437-442.
1. Malizos KN, Quarles LD, Seaber AV, et al. An experimental canine model of osteonecrosis:characterization of the repair process. J Orthop Res,1993; 11(3): 350-357.
2. Mont MA, Jones LC, Hungerford DS. Nontraumatic osteonecrosis of the femoral head: ten years later.J Bone Joint Surg Am.2006; 88(5):1117-1132.
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