中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (28): 5950-5956.doi: 10.12307/2025.479

• 组织工程骨材料 tissue-engineered bone • 上一篇    下一篇

骨诱导膜中H型血管动态表达及耦合骨生成修复大段骨缺损

申  震，黄梓越，和智娟，王艺婷，陈奇刚，耿春梅，黄雅静，吴祖贵   

  1. 云南中医药大学第三附属医院(昆明市中医医院)，云南省昆明市   650011
  • 收稿日期:2024-04-08 接受日期:2024-07-20 出版日期:2025-10-08 发布日期:2024-12-07
  • 通讯作者: 黄雅静，护师，云南中医药大学第三附属医院(昆明市中医医院)风湿肾病科，云南省昆明市 650011 吴祖贵，博士，医师，云南中医药大学第三附属医院(昆明市中医医院)骨伤科，云南省昆明市 650011
  • 作者简介:申震，男，1989年生，江苏省徐州市人，汉族，博士，主治医师，主要从事中医药防治骨与关节损伤方面的研究。
  • 基金资助:
    国家自然科学基金项目(82360943)，项目负责人：申震；云南省“兴滇英才”支持计划青年人才专项项目(XDYC-QNRC-2022-0609)，项目负责人：申震；云南省基础研究专项基金项目(202201AU070120)，项目负责人：申震；云南省中医联合专项基金项目(202101AZ070001-123，2021AZ070001-257)，项目负责人：申震；昆明市卫生科技人才培养项目曁“十百千”工程项目(2021-SW-52)，项目负责人：申震

Dynamic expression of H-type vessels coupled with bone repair effect in bone induced membrane for massive bone defects

Shen Zhen, Huang Ziyue, He Zhijuan, Wang Yiting, Chen Qigang, Geng Chunmei, Huang Yajing, Wu Zugui   

  1. Third Affiliated Hospital (Kunming Traditional Chinese Medicine Hospital) of Yunnan University of Chinese Medicine, Kunming 650011, Yunnan Province, China
  • Received:2024-04-08 Accepted:2024-07-20 Online:2025-10-08 Published:2024-12-07
  • Contact: Huang Yajing, Senior nurse, Third Affiliated Hospital (Kunming Traditional Chinese Medicine Hospital) of Yunnan University of Chinese Medicine, Kunming 650011, Yunnan Province, China Wu Zugui, MD, Physician, Third Affiliated Hospital (Kunming Traditional Chinese Medicine Hospital) of Yunnan University of Chinese Medicine, Kunming 650011, Yunnan Province, China
  • About author:Shen Zhen, MD, Attending physician, Third Affiliated Hospital (Kunming Traditional Chinese Medicine Hospital) of Yunnan University of Chinese Medicine, Kunming 650011, Yunnan Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 82360943 (to SZ); “Xingdianyingcai” Young Talent Support Project of Yunnan Province, No. XDYC-QNRC-2022-0609 (to SZ); Basic Research Project of Science and Technology Department of Yunnan Province, No. 202201AU070120 (to SZ); Yunnan Province Traditional Chinese Medicine Joint Special Fund Project, No. 202101AZ070001-123, 2021AZ070001-257 (to SZ); Kunming Health Science and Technology Talent Cultivation Project and “Ten Hundred Thousand” Talent Project, No. 2021SW52 (to SZ)

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摘要：


文题释义：

Masquelet 技术：又称骨诱导膜技术，是法国医师Masquelet于1986年首次提出的一种骨再生技术，分2个阶段：第一阶段，先经过彻底清创，将骨水泥填充在骨缺损处，防止周围软组织嵌入后阻碍骨修复；第二阶段，第一阶段术后 6-8周，在骨缺损处骨水泥周围生成完整诱导膜，切开诱导膜，在保持诱导膜相对完整的情况下取出骨水泥，进行骨移植手术。经过数十年的发展，骨诱导膜技术被广泛应用于肢体形态与功能重建领域，并获得了较好疗效。
H型血管：是存在于骨骼系统血管网络末端的一种特殊的毛细血管亚型，对血小板内皮细胞黏附分子CD31和唾液糖蛋白内黏蛋白Endomucin(Emcn)同时高表达(CD31hiEmcnhi)，在形态、功能和空间分布方面具有特异性，在骨骼生长发育、修复重建和维持骨量平衡过程中具有分泌因子、信号传递、功能调控等重要作用。


背景：骨诱导膜技术治疗大段骨缺损存在骨修复缓慢、成骨质量不佳等问题。H型血管具有诱导成骨作用，可增强局部成血管-成骨偶联作用，促进骨修复，但H型血管在骨诱导膜中的作用鲜有报道。

目的：构建SD大鼠胫骨大段骨缺损模型，观察骨诱导膜中H型血管表达特征，进而明确骨诱导膜中H型血管表达高峰点与植骨最佳时期。
方法：采用随机数字表法将60只SD大鼠随机分为对照组(n=30)与模型组(n=30)，两组又各自分为骨水泥植入后4，6，8周3个亚组，每组10只大鼠。对照组与模型组大鼠均构建右侧胫骨4 mm骨缺损模型，模型组植入聚甲基丙烯酸甲酯骨水泥诱导骨生物膜形成，对照组不植入骨水泥。骨水泥植入后4，6，8周，每个时间点随机取6只大鼠，模型组切取骨诱导膜组织，对照组切取对应部位的非骨性软组织，通过免疫荧光鉴定H型血管动态表达，苏木精-伊红染色观察诱导膜组织形态改变，血管造影观察血管形成情况，免疫组化染色检测成骨细胞特异性转录因子表达；每个时间点剩余4只大鼠，模型组切开诱导膜后取出骨水泥，植入自体尾骨，对照组骨缺损区植入自体尾骨，植骨后8周进行胫骨缺损部位Micro-CT评估。

结果与结论：①免疫荧光染色显示，模型组H型血管表达在骨水泥植入后6周最明显，模型组骨水泥植入后各时间点的H型血管表达高于对照组(P < 0.05)；②苏木精-伊红染色与血管造影检测显示，模型组骨水泥植入后各时间点的新生血管数、血管体积均大于对照组(P < 0.05)，模型组组内新生血管数、血管体积大小顺序为：骨水泥植入后8周>骨水泥植入后6周>骨水泥植入后4周；③免疫组化染色显示，模型组骨水泥植入后各时间点的成骨细胞特异性转录因子阳性表达高于对照组(P < 0.05)，模型组成骨细胞特异性转录因子阳性表达在骨水泥植入后6周最明显；④Micro-CT检测显示，模型组3个亚组的骨修复效果明显优于对照组对应的亚组，模型组骨水泥植入后6周亚组的骨修复效果优于骨水泥植入后4，8周亚组。结果表明：骨诱导膜中H型血管呈动态表达并在骨水泥植入后6周达高峰，在骨水泥植入后6周进行骨诱导膜植骨可获得良好的骨修复效果。

https://orcid.org/0000-0002-6109-9775 (申震) 

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

关键词: Masquelet技术, 骨诱导膜, H型血管, 动态表达, 骨修复, 大段骨缺损, 骨水泥, 工程化骨材料

Abstract: BACKGROUND: Slow bone repair and poor bone formation quality are still problems during masquelet technique in the treatment of large segment bone defects. H-type vessels can induce osteogenesis, enhance the local angiogenesis and osteogenesis coupling, and promote bone repair. However, there are few reports on the role of H-type blood vessels in the bone induced membrane. 
OBJECTIVE: To construct a large segment bone defect model of SD rat tibia, observe the expression characteristics of H-type blood vessels in the bone induced membrane, then to identify the expression peak point of H-type blood vessels in the bone induced membrane and determine the optimal period of bone grafting.
METHODS: Sixty SD rats were randomly divided into a control group (n=30) and a model group (n=30) by random number table method. The two groups were further divided into three subgroups at 4, 6, and 8 weeks after bone cement implantation, with 10 rats in each group. A 4 mm bone defect model of the right tibia was constructed in both the control and the model groups. Polymethyl methacrylate bone cement was implanted in the model group to induce bone biomembrane formation, while bone cement was not implanted in the control group. At 4, 6, and 8 weeks after bone cement implantation, 6 rats were randomly selected at each time point. The bone induction membrane tissue was cut from the model group, and the non-bone soft tissue of the corresponding part was cut from the control group. The dynamic expressions of H-type blood vessels in the bone induced membrane were identified by immunofluorescence. The morphological changes of the bone induced membrane were observed by hematoxylin-eosin staining. The formation of blood vessels in the bone induced membrane was observed by angiography. The expression levels of osteoblast-specific transcription factor in the bone induced membrane were detected by immunohistochemistry. Four rats remained at each time point. In the model group, the bone induced membrane was cut open and the bone cement was removed and autologous coccyx was implanted. In the control group, autologous coccyx was implanted in the bone defect area. Micro-CT evaluation of the tibial defect was performed 8 weeks after bone grafting. 
RESULTS AND CONCLUSION: (1) Immunofluorescence staining showed that the expression of H-type vessels in the model group was most obvious 6 weeks after bone cement implantation, and the expression of H-type vessels in the model group at each time point after bone cement implantation was higher than that in the control group (P < 0.05). (2) Hematoxylin-eosin staining and angiography showed that the number and volume of new blood vessels at each time point after bone cement implantation in the model group were greater than those in the control group (P < 0.05). The order of the number and volume of new blood vessels in the model group was: 8 weeks after bone cement implantation > 6 weeks after bone cement implantation > 4 weeks after bone cement implantation. (3) Immunohistochemical staining showed that the positive expression of osteoblast-specific transcription factors at each time point after bone cement implantation in the model group was higher than that in the control group (P < 0.05), and the positive expression of osteoblast-specific transcription factors in the model group was most obvious 6 weeks after bone cement implantation. (4) Micro-CT detection showed that the bone repair effect of the three subgroups in the model group was significantly better than that of the corresponding subgroups in the control group, and the bone repair effect of the subgroup in the model group 6 weeks after bone cement implantation was better than that of the subgroups 4 and 8 weeks after bone cement implantation. The results indicate that H-type blood vessels are dynamically expressed in the bone induced membrane and reached a peak 6 weeks after bone cement implantation. Good bone repair effects can be obtained by the bone induced membrane bone grafting 6 weeks after bone cement implantation. 

Key words: Masquelet technique, bone induced membrane, H-type vessels, dynamic expression, bone repair, massive bone defect, bone cement, engineered bone material

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