Automatic Quantitative Micro-Computed Tomography Evaluation of Angiogenesis in an Axially Vascularized Tissue-Engineered Bone Construct
Automatic Quantitative Micro-Computed Tomography Evaluation of Angiogenesis in an Axially Vascularized Tissue-Engineered Bone Construct
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DOI：
10.1089/ten.tec.2010.0016
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发表时间：
2010-12-01
影响因子：
3
通讯作者：
Kneser, Ulrich
中科院分区：
文献类型：
--
作者：
Arkudas, Andreas;Beier, Justus Patrick;Kneser, Ulrich
简介：我们发明了一种独立于观察者的自动定量方法，利用显微计算机断层扫描（CT）和组织工程模型的三维重建来分析血管化。材料与方法：在30只大鼠大腿内侧建立动静脉环，置于多孔羟基磷灰石和β -磷酸三钙颗粒基质中，填充纤维蛋白（10mg/mL纤维蛋白原和2iu /mL凝血酶），不加（a组）或加（B组）纤维蛋白凝胶固定血管生成生长因子血管内皮生长因子（VEGF(165)）和碱性成纤维细胞生长因子（bFGF）。外植间隔分别为2周、4周和8周。通过显微ct对标本进行调查，然后进行自动3D分析，这与组织形态学结果相关。结果：在两组中，动静脉环导致基质内血管分化和功能血管的致密化结缔组织的产生。显微ct对血管形成的定量分析优于组织学分析。micro-CT分析还允许在3D构建中评估不同的其他更复杂的血管化参数，表明纤维蛋白凝胶固定化VEGF（165）和bFGF的应用可以早期改善血管化。结论：在本研究中，利用显微ct定量分析血管形成，并结合三维重建和自动分析，是一种优于横截面组织学评估的有效方法。
Introduction: We invented an automatic observer-independent quantitative method to analyze vascularization using micro-computed tomography (CT) along with three-dimensional (3D) reconstruction in a tissue engineering model.Materials and Methods: An arteriovenous loop was created in the medial thigh of 30 rats and was placed in a particulated porous hydroxyapatite and beta-tricalcium phosphate matrix, filled with fibrin (10mg/mL fibrinogen and 2 IU/mL thrombin) without (group A) or with (group B) application of fibrin-gel-immobilized angiogenetic growth factors vascular endothelial growth factor (VEGF(165)) and basic fibroblast growth factor (bFGF). The explantation intervals were 2, 4, and 8 weeks. Specimens were investigated by means of micro-CT followed by an automatic 3D analysis, which was correlated to histomorphometrical findings.Results: In both groups, the arteriovenous loop led to generation of dense vascularized connective tissue with differentiated and functional vessels inside the matrix. Quantitative analysis of vascularization using micro-CT showed to be superior to histological analysis. The micro-CT analysis also allows the assessment of different other, more complex vascularization parameters within 3D constructs, demonstrating an early improvement of vascularization by application of fibrin-gel-immobilized VEGF(165) and bFGF.Conclusions: In this study quantitative analysis of vascularization using micro-CT along with 3D reconstruction and automatic analysis exhibit to be a powerful method superior to histological evaluation of cross sections.