Karpenko A A, Rozanova I A, Poveshchenko O V, Lykov A P, Bondarenko N A, Kim I I, Nikonorova Iu V, Podkhvatilina N A, Sergeevichev D S, Popova I V, Konenkov V I
Laboratory of Experimental Surgery and Morphology, Centre for Vascular and Hybrid Surgery, Novosibirsk Scientific Research Institute for Circulatory Pathology named after Academician E.N. Meshalkin under the RF Ministry of Public Health, Novosibirsk, Russia.
Laboratory of Experimental Surgery and Morphology, Centre for Vascular and Hybrid Surgery, Novosibirsk Scientific Research Institute for Circulatory Pathology named after Academician E.N. Meshalkin under the RF Ministry of Public Health, Novosibirsk, Russia; Laboratory of Cellular Technologies, Scientific Research Institute of Clinical and Experimental Lymphology under the Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
Angiol Sosud Khir. 2015;21(4):178-84.
Search for new bioengineering materials for creation of small-diameter vascular grafts is currently a priority task. One of the promising trends of creating tissue engineering constructions is coating the internal layer of implants made of polytetrafluoroethylene (PTFE) with autologous mesenchymal multipotent stromal cells. In the study we assessed the ability of separate components of the extracellular matrix such as fibronectin, type I collagen and type IV collagen to influence adhesion, proliferation and morphology of mesenchymal multipotent stromal cells being cultured on PTFE. Bone marrow multipotent stromal cells taken from second-passage Wistar rats in the amount of 106 per 1 cm2 were applied onto PTFE. We used the following variants of preliminary treatment of the material prior to seeding: fibronectin with type I collagen, fibronectin with type IV collagen, fibronectin with a mixture of type I and IV collagens, as well as a control group without coating. After six weeks of cell growth on PTFE patches the samples were subjected to fixation in 10% formalin followed by haematoxylin-eosin stain and morphometric assessment of adhered cells by calculation using the software AxioVision (Carl Zeiss), assessing the number of cells, area of the cellular monolayer, dimensions and ratios of the area of separate cells and the area of cellular nuclei. The maximal area of the monolayer from mesenchymal multipotent stromal cells on the PTFE surface was revealed while culturing with a mixture of fibronectin and type I and IV collagens. Cell colonization density while treatment of the synthetic material with mixtures of fibronectin with type I collagen, type IV collagen and type I and IV collagens demonstrated the results exceeding the parameters of the control specimen 5-, 2.5- and 7-fold, respectively. Hence, extracellular matrix components considerably increase enhance adhesion of cells to PTFE, as well as improve formation of a monolayer from mesenchymal multipotent stromal cells.
寻找用于制造小口径血管移植物的新型生物工程材料是当前的一项优先任务。创建组织工程结构的一个有前景的趋势是用自体间充质多能基质细胞覆盖由聚四氟乙烯（PTFE）制成的植入物内层。在该研究中，我们评估了细胞外基质的单独成分，如纤连蛋白、I型胶原蛋白和IV型胶原蛋白，对在PTFE上培养的间充质多能基质细胞的黏附、增殖和形态的影响。从第二代Wistar大鼠获取的骨髓多能基质细胞，以每1平方厘米106个的数量接种到PTFE上。在接种前，我们使用了以下材料预处理变体：纤连蛋白与I型胶原蛋白、纤连蛋白与IV型胶原蛋白、纤连蛋白与I型和IV型胶原蛋白的混合物，以及未涂层的对照组。在PTFE贴片上细胞生长六周后，将样本用10%福尔马林固定，然后进行苏木精-伊红染色，并使用AxioVision软件（卡尔·蔡司公司）通过计算对黏附细胞进行形态计量评估，评估细胞数量、细胞单层面积、单个细胞面积与细胞核面积的尺寸及比例。在用纤连蛋白与I型和IV型胶原蛋白的混合物培养时，PTFE表面的间充质多能基质细胞形成的单层面积最大。在用纤连蛋白与I型胶原蛋白、IV型胶原蛋白以及I型和IV型胶原蛋白的混合物处理合成材料时，细胞定植密度分别比对照样本的参数高出5倍、2.5倍和7倍。因此，细胞外基质成分显著增强了细胞与PTFE的黏附，并改善了间充质多能基质细胞单层的形成。
