Gille J, Meisner U, Ehlers E M, Müller A, Russlies M, Behrens P
Department of Orthopaedic Surgery, University of Schleswig-Holstein, Campus Lübeck, Ratzeburger Allee 160 D-23538, Germany.
Tissue Cell. 2005 Oct;37(5):339-48. doi: 10.1016/j.tice.2005.05.004.
We studied the migration pattern, morphology and viability of cells suspended in five different fibrin glues. Besides this, the behaviour of chondrocytes seeded on porous matrices comprising different collagen types sealed with fibrin glue was investigated.
In an experiment A, cell suspension (0.5x10(6) cells) was incubated with different fibrin glues. Experiment B was set up to evaluate chondrocytes migration either through a collagen I/III (Chondro-Gide, Geistlich Biomaterials, Switzerland) or collagen II matrix sealed with different fibrin glues in a perfusion chamber system. Analysis were performed by lightmicroscopy (Mayer's hematoxylin-eosin; Masson-Goldner; TUNEL test) and by transmission and scanning electron microscopy. All fibrin glues were measured for TGF-beta 1 and 2 with a specific ELISA.
After incubation of cell suspension in autologous fibrin glue, the morphology of cells is chondrocyte-like. Spindly, process-bearing cells were seen in commercial fibrin glue. Cells suspended in commercial fibrin glue revealed a significant higher percentage of TUNEL positive cells compared to fibrin tissue adhesives mixed with autologous serum (p=0.006). The TGF-beta 1 and 2 concentration was significantly higher in partial autologous fibrin sealant (PAF) compared to their commercial counterparts (p=0.001). Cells seeded on the collagen I/III matrix retained their chondrocytic morphology, while in the type II collagen matrix the chondrocytes displayed a fibroblastic phenotype. The ratio of TUNEL positive cells for the collagen I/III matrix was significantly surpassed by the values, when a collagen II matrix was used (p=0.008). No ingrowth of cells was seen in any of the experimental conditions.
Partial autologous fibrin glue and collagen I/III matrices are favourable in respect to migration pattern, morphology and viability, but definitive conclusions can only be drawn after in vivo studies. This will be addressed in future animal studies.
我们研究了悬浮于五种不同纤维蛋白胶中的细胞的迁移模式、形态及活力。此外，还研究了接种在由不同类型胶原蛋白构成并用纤维蛋白胶封闭的多孔基质上的软骨细胞的行为。
在实验A中，将细胞悬液（0.5×10⁶个细胞）与不同的纤维蛋白胶一起孵育。实验B旨在评估软骨细胞在灌注室系统中通过用不同纤维蛋白胶封闭的I/III型胶原蛋白（Chondro - Gide，Geistlich生物材料公司，瑞士）或II型胶原蛋白基质的迁移情况。通过光学显微镜（苏木精 - 伊红染色；马松 - 戈德纳染色；TUNEL检测）以及透射和扫描电子显微镜进行分析。使用特异性酶联免疫吸附测定法检测所有纤维蛋白胶中的转化生长因子 - β1和 - β2。
细胞悬液在自体纤维蛋白胶中孵育后，细胞形态呈软骨细胞样。在商用纤维蛋白胶中可见细长的、有突起的细胞。与混合自体血清的纤维蛋白组织粘合剂相比，悬浮于商用纤维蛋白胶中的细胞显示出TUNEL阳性细胞的比例显著更高（p = 0.006）。部分自体纤维蛋白密封剂（PAF）中的转化生长因子 - β1和 - β2浓度显著高于其商用对应物（p = 0.001）。接种在I/III型胶原蛋白基质上的细胞保持其软骨细胞形态，而在II型胶原蛋白基质中软骨细胞呈现成纤维细胞表型。当使用II型胶原蛋白基质时，其TUNEL阳性细胞的比例显著超过I/III型胶原蛋白基质的值（p = 0.008）。在任何实验条件下均未见细胞向内生长。
部分自体纤维蛋白胶和I/III型胶原蛋白基质在迁移模式、形态及活力方面是有利的，但只有在体内研究后才能得出明确结论。这将在未来的动物研究中进行探讨。
