诱导膜在骨修复中的作用机制.docx

The Mechanism of Action of Induced Membranes in Bone Repair.诱导膜在骨修复中的作用机制AbstractBACKGROUND:Inducement of foreign-body granulation tissue is a relatively novel therapeutic modality in bone repair. A two-stage bone reconstruction method, known as the Masquelet technique, combines inducement of a granulation tissue membrane and subsequent bone autografting as a biphasic technique allowing reconstruction of large bone defects. In light of their already well-characterized osteogenesis-improving capabilities in animals, we performed this translational study to investigate these membranes in patients.摘要背景：诱导异物肉芽组织在骨修复中是一种较新的治疗方法。两阶段骨重建方法，即熟知的Masquelet技术，是结合诱导肉芽组织膜和随后的骨自体移植的双向技术以允许大段骨缺损重建。根据它们在动物实验中良好的提高成骨能力特点，我们将这些膜植入患者中进行研究。METHODS:Fourteen patients with complicated fractures and bone defects were randomly selected for this study. Biopsy samples of foreign-body-induced membranes were collected at different time points during scheduled surgical procedures. The membranes were co-cultured with mesenchymal stromal cells, and differentiation into the osteoblastic lineage was assessed by measuring alkaline phosphatase activity, aminoterminal propeptide of type-I procollagen (PINP) production, and Ca2+ concentration. Histological characteristics were evaluated with image analysis. Quantitative reverse transcription polymerase chain reaction was used to measure vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), and type-I collagen (Col-1) expression.方法：本研究将14例复杂骨折并骨缺损的患者随机分组。按预定的手术方法在不同时间点活检收集异物诱导的膜。这些膜和间充质干细胞共同培养，通过测定碱性磷酸酶活性、I型胶原（PINP）产物氨基端前肽、以及钙离子浓度评估其分化成成骨细胞谱系。用图像分析对组织学特点进行评价。应用定量逆转录聚合酶链反应法对血管内皮生子因子(VEGF)、白细胞介素-6 (IL-6)，以及I型胶原蛋白(Col-1)表达进行测量。RESULTS:The induced membranes were characterized histologically by maturating vascularized fibrous tissue. The vascularization was greatest in one-month-old samples and decreased to 60% in three-month-old samples. One-month-old membrane samples had the highest expression of VEGF, IL-6, and Col-1, whereas two-month-old membranes expressed 40% of the levels of the one-month-old membranes. Specific alkaline phosphatase activity, PINP production, and Ca2+ concentration were increased in co-cultures when a membrane sample was present. In cultures of one-month-old membranes, PINP production was more than two times and Ca2+ deposition was four times higher than that in cultures of two-month-old membranes.结果：诱导的膜具有成熟的血管纤维组织特点。血管化在一个月的的标本中程度最高，在三个月的标本中减少至少于60。一个月膜标本中VEGF、IL-6以及Col-1表达最高，而在两个月膜标本中这些因子表达不到一个月膜标本的40。特异碱性磷酸酶活性、PINP产物以及钙离子浓度在膜存在共同培养时增高。在培养中一个月的膜标本较两个月的膜标本，PINP产物增高两倍多，钙离子浓度增高四倍。CONCLUSIONS:The induced membranes have osteogenesis-improving capabilities. These capabilities, however, appear to decrease over time. We speculate that the optimal time for performing second-stage surgery may be within a month after implantation of foreign material.结果：诱导的膜具有提高成骨的能力。然而这种能力似乎随着时间减弱。我们推测在异物植入后一个月内可能是进行第二阶段手术的最佳时间。CLINICAL RELEVANCE:The two-stage bone reconstruction method is an easily adaptable technique for repair of large bone defects. Further understanding of membrane biology and matura