| 瓦楞状组织工程骨支架对种子细胞接种和成骨的影响 |
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| 引用本文： | 曾宪利,杨春露,李 江,徐 湲,王 硕,吴 鹏,王翠芳,丁艳丽,郑新丽. 瓦楞状组织工程骨支架对种子细胞接种和成骨的影响[J]. 中国组织工程研究, 2014, 18(25): 3953-3960. DOI: 10.3969/j.issn.2095-4344.2014.25.004 |
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| 作者姓名： | 曾宪利 杨春露 李 江 徐 湲 王 硕 吴 鹏 王翠芳 丁艳丽 郑新丽 |
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| 作者单位： | 1沈阳医学院附属中心医院，辽宁省手外科组织工程实验室，辽宁省沈阳市 110024；2武警辽宁省总队医院骨科，辽宁省沈阳市 110034；3沈阳医学院科学实验中心，辽宁省沈阳市 110034 |
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| 基金项目： | 辽宁省手外科组织工程实验室建设项目(201104523)；辽宁省科技厅自然科技基金(201102217)；沈阳医学院优秀人才基金(20081024) |
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| 摘 要： | 背景：不同形状工程骨支架负载种子细胞修复骨缺损的研究效果评价不一，而负载细胞数量的多少是影响疗效的重要因素之一，目前该方面研究证据不多。目的：自制瓦楞状组织工程骨支架和其他3种形状的支架，比较4种不同形状支架负载种子细胞的数量，以及瓦楞状组织工程骨支架体内成骨时凹槽的优势及特点。方法：①体外实验：将体积和样本数相同的4组支架分为单纯瓦楞状支架组、无瓦楞支架组、圆柱状支架组和带中空管柱状支架组，分别以相同密度、相同容积的成骨诱导兔骨髓间充质干细胞悬液接种于支架表面，孵育、培养、消化、收集，进行细胞计数、吸光度值检测以及碱性磷酸酶和茜素红染色。②体内实验：将兔随机分为重组人骨形态发生蛋白2/瓦楞状自固化磷酸钙人工骨组，瓦楞状自固化磷酸钙人工骨组和松质骨组，将体积相同的3组支架植入兔L5-6两侧横突间，植入后4，8，12周行大体、组织学观察。结果与结论：体外实验显示，瓦楞状工程骨支架上滴注的细胞液能充分停留在表面，由于表面瓦楞状凹槽和液体的表面张力以及支架本身的无孔隙性使得细胞液不向培养皿流失，每个样本平均消化下来的正常形态的细胞数量高于其他3组(P < 0.05)，吸光度值差异无显著性意义(P > 0.05)。体内实验显示，各时间点重组人骨形态发生蛋白2/瓦楞状自固化磷酸钙人工骨组的成骨量比瓦楞状自固化磷酸钙人工骨组明显多(P < 0.05)，与松质骨组之间比较差异无显著性意义(P > 0.05)。结果证实，实验自制的瓦楞状工程骨支架的形状特点有利于种子细胞负载，从数量上保证了接种种子细胞的有效性，可促进支架大量成骨和段状骨缺损的愈合。中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程全文链接：
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| 关 键 词： | 生物材料 骨生物材料 瓦楞状 骨组织构建 自固化磷酸钙人工骨 重组人骨形态发生蛋白2 凹槽 组织工程骨支架 骨髓间充质干细胞 接种 静态接种法 成骨 兔 |
| 收稿时间： | 2014-05-16 |
Effects of corrugated tissue engineered bone scaffold on cell seeding and osteogenesis |
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| Zeng Xian-li,Yang Chun-lu,Li Jiang,Xu Yuan,Wang Shuo,Wu Peng,Wang Cui-fang,Ding Yan-li,Zheng Xin-li. Effects of corrugated tissue engineered bone scaffold on cell seeding and osteogenesis[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(25): 3953-3960. DOI: 10.3969/j.issn.2095-4344.2014.25.004 |
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| Authors: | Zeng Xian-li Yang Chun-lu Li Jiang Xu Yuan Wang Shuo Wu Peng Wang Cui-fang Ding Yan-li Zheng Xin-li |
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| Affiliation: | 1Tissue Engineering Laboratory of Hand Surgery of Liaoning Province, Central Hospital Affiliated to Shenyang Medical College, Shenyang 110024, Liaoning Province, China; 2Department of Orthopedics, Armed Police Corps General Hospital in Liaoning Province, Shenyang 110034, Liaoning Province, China; 3Science Experiment Center of Shenyang Medical College, Shenyang 110034, Liaoning Province, China |
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| Abstract: | BACKGROUND:The effects of engineered bone scaffold containing seeding cells with different shapes to repair bone defect are varied, while the loaded cell quantity is the important factor influencing the curative effect, but which is rarely reported. OBJECTIVE: By preparing self-made corrugated tissue-engineered bone scaffold and other three forms of bone tissue engineering scaffolds, to study the quantity of loaded cells on different scaffolds and osteogenesis of corrugated tissue-engineered bone scaffold so as to discuss the advantages and features of self-made corrugated tissue-engineered bone scaffold.METHODS: (1) Experiment in vitro: There were four kinds of scaffolds with the same volume and samples: calcium phosphate cement (CPC) corrugated surface scaffold group, smooth surface scaffold group, cylindrical scaffold group and porous cylindrical scaffold with hollow tubes group, in which the latter three groups are control ones. A certain volume with same density of rabbit bone marrow mesenchymal stem cells (BMSCs) suspension after osteogenesis induction was seeded onto the scaffolds. After incubation, culture, digestion and collection, cell quantity was counted, absorbance value was finally detected and cell activity was proofed by alkaline phosphatase and alizarin red staining. (2) Experiment in vivo: New Zealand rabbits were randomly and equally divided into recombinant human bone morphogenetic protein-2 (rhBMP-2)/CPC/BMSCs corrugated scaffold group, pure CPC corrugated scaffold group and cancellous bone implant group. Three kinds of scaffold implants with the same volume were inserted into the area between rabbit’s L5, 6 transverse processes bilaterally. At 4, 8, 12 weeks postoperatively, gross and histological observation was performed. RESULTS AND CONCLUSION:(1) Experiment in vitro: The drip of cell suspension steadily stayed on the surface of corrugated scaffold because of corrugated shape groove and the surface tension of the liquid. The amount of cells per sample digested down from the CPC corrugated surface scaffold was significantly more than that from the other three groups (P < 0.05), while the absorbance values did not differ (P > 0.05). (2) Experiment in vivo: At each time point the osteogenesis quantity of rhBMP-2/CPC/BMSCs corrugated scaffold group was more than that of the pure CPC corrugated scaffold group (P < 0.05), while there was no difference from the cancellous bone implant group (P > 0.05). These findings indicate that the characteristics of the self-made corrugated engineered bone scaffold are beneficial to seed cell loading, which supports a large number of osteogenesis and provides feasibility to promote the healing of segmental bone defects. |
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| Keywords: | calcium phosphates bone morphogenetic proteins stents mesenchymal stem cells |
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