Schumann Paul, Tavassol Frank, Lindhorst Daniel, Stuehmer Constantin, Bormann Kai-Hendrik, Kampmann Andreas, Mülhaupt Rolf, Laschke Matthias W, Menger Michael D, Gellrich Nils-Claudius, Rücker Martin
Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Strasse 1, D-30625 Hannover, Germany.
Microvasc Res. 2009 Sep;78(2):180-90. doi: 10.1016/j.mvr.2009.06.003. Epub 2009 Jun 21.
Implantation of tissue engineering constructs is a promising technique to reconstruct injured tissue. However, after implantation the nutrition of the constructs is predominantly restricted to vascularization. Since cells possess distinct angiogenic potency, we herein assessed whether scaffold vitalization with different cell types improves scaffold vascularization. 32 male balb/c mice received a dorsal skinfold chamber. Angiogenesis, microhemodynamics, leukocyte-endothelial cell interaction and microvascular permeability induced in the host tissue after implantation of either collagen coated poly (L-lactide-co-glycolide) (PLGA) scaffolds (group 4), additionally seeded with osteoblast-like cells (OLCs, group 1), bone marrow mesenchymal stem cells (bmMSCs, group 2) or a combination of OLCs and bmMSCs (group 3) were analyzed repetitively over 14 days using intravital fluorescence microscopy. Apart from a weak inflammatory response in all groups, vascularization was found distinctly accelerated in vitalized scaffolds, indicated by a significantly increased microvascular density (day 6, group 1: 202+/-15 cm/cm(2), group 2: 202+/-12 cm/cm(2), group 3: 194+/-8 cm/cm(2)), when compared with controls (group 4: 72+/-5 cm/cm(2)). This acceleration was independent from the seeded cell type. Immunohistochemistry revealed in vivo VEGF expression in close vicinity to the seeded OLCs and bmMSCs. Therefore, the observed lack of cell type confined differences in the vascularization process suggests that the accelerated vascularization of vitalized scaffolds is VEGF-related rather than dependent on the potential of bmMSCs to differentiate into specific vascular cells.
组织工程构建体的植入是一种重建受损组织的有前景的技术。然而,植入后构建体的营养主要限于血管化。由于细胞具有不同的血管生成能力,我们在此评估用不同细胞类型对支架进行活化是否能改善支架的血管化。32只雄性balb/c小鼠接受了背部皮褶腔室。在植入胶原包被的聚(L-丙交酯-共-乙交酯)(PLGA)支架(第4组)、另外接种成骨样细胞(OLCs,第1组)、骨髓间充质干细胞(bmMSCs,第2组)或OLCs与bmMSCs的组合(第3组)后,使用活体荧光显微镜在14天内重复分析宿主组织中诱导的血管生成、微循环动力学、白细胞-内皮细胞相互作用和微血管通透性。除了所有组中均有微弱的炎症反应外,与对照组(第4组:72±5个/cm/cm²)相比,活化支架中的血管化明显加速,表现为微血管密度显著增加(第6天,第1组:202±15个/cm/cm²,第2组:202±12个/cm/cm²,第3组:194±8个/cm/cm²)。这种加速与接种的细胞类型无关。免疫组织化学显示在接种的OLCs和bmMSCs附近有体内VEGF表达。因此,在血管化过程中观察到的缺乏细胞类型限制的差异表明,活化支架的血管化加速与VEGF相关,而不是依赖于bmMSCs分化为特定血管细胞的潜能。