Wang Xiaohua, Wei Fuda, Luo Fei, Huang Ke, Xie Zhao
National & Regional United Engineering Laboratory of Tissue Engineering, Department of Orthopaedics, Southwest Hospital, Third Military Medical University, Chongqing, 400038, The People's Republic of China.
J Orthop Surg Res. 2015 Sep 17;10:147. doi: 10.1186/s13018-015-0287-4.
The use of the Masquelet technique in the repair of large bone defects has gained increased acceptance in recent years. The core of this technique is the induction of granulation tissue membrane formation and the implantation of an autologous cancellous bone to reconstruct bone defects in the membrane. In this study, we purpose to explore the structure of induced membrane and the content of growth factors as well to compare between the structure and the effects on osteogenesis of induced membranes and the periosteum in animal models.
Bilateral radial bone defects were generated in 32 healthy adult rabbits. The defects were implanted with bone cement. The induced membranes and periosteum were removed after 2, 4, 6, and 8 weeks. Thereafter, hematoxylin-eosin staining (HE) and an enzyme-linked immunosorbent assay (ELISA) were performed to detect vascular endothelial growth factor (VEGF), angiotensin II (ANG-II), bone morphogenetic protein 2 (BMP2), fibroblast growth factor 2 (FGF2), and prostaglandin E2 (PGE2). Proteins isolated from total cell lysates were cultured with mesenchymal stem cells to test the cell proliferation and alkaline phosphatase activity using epimysium as a control.
The induced membrane and periosteum exhibited similar structures and growth factor levels after 4 and 6 weeks. The highest concentration of BMP-2 and VEGF in the induced membranes occurred in week 6, and FGF-2 and ANG-II concentrations peaked in week 4. The thickness and vascular density of induced membranes gradually decreased with time.
Induced membrane matured between the 4th and the 6th week and secreted growth factors to promote osteogenesis. The matured induced membrane and periosteum had similar structures and abilities to promote the osteogenesis of mesenchymal stem cells. However, the induced membrane was thicker than the periosteum.
近年来，Masquelet技术在大骨缺损修复中的应用越来越受到认可。该技术的核心是诱导肉芽组织膜形成，并植入自体松质骨以重建膜内的骨缺损。在本研究中，我们旨在探索诱导膜的结构和生长因子含量，并在动物模型中比较诱导膜和骨膜的结构及其对成骨的影响。
在32只健康成年兔中制造双侧桡骨缺损。缺损处植入骨水泥。在2、4、6和8周后取出诱导膜和骨膜。此后，进行苏木精-伊红染色（HE）和酶联免疫吸附测定（ELISA），以检测血管内皮生长因子（VEGF）、血管紧张素II（ANG-II）、骨形态发生蛋白2（BMP2）、成纤维细胞生长因子2（FGF2）和前列腺素E2（PGE2）。从总细胞裂解物中分离的蛋白质与间充质干细胞一起培养，以使用肌外膜作为对照测试细胞增殖和碱性磷酸酶活性。
诱导膜和骨膜在4周和6周后表现出相似的结构和生长因子水平。诱导膜中BMP-2和VEGF的最高浓度出现在第6周，FGF-2和ANG-II浓度在第4周达到峰值。诱导膜的厚度和血管密度随时间逐渐降低。
诱导膜在第4至6周成熟并分泌生长因子以促进成骨。成熟的诱导膜和骨膜具有相似的结构和促进间充质干细胞成骨的能力。然而，诱导膜比骨膜厚。
