Bücheler M, Scheffler B, von Foerster U, Bruinink A, Bootz F, Wintermantel E
Klinik und Poliklinik für Hals-Nasen-Ohrenheilkunde, Plastische Operationen, Universität Leipzig.
Laryngorhinootologie. 2000 Mar;79(3):160-4. doi: 10.1055/s-2000-286.
Clinical application of bioartificial tracheal prosthesis must still be regarded as an experimental concept because restoration of a functional respiratory epithelium outlining the prosthesis is still not possible. Tissue engineering as a relatively new biotechnological discipline may offer new methods in expanding differentiated respiratory epithelium in vitro. In this study we compare two different cell and tissue culture procedures for growing human nasal mucosa on commercially available collagen foil.
Harvested specimens of human nasal mucosa (n = 6, 4 x 4 cm) were placed on collagen foil and incubated as tissue cultures for 4, 6 and 8 weeks. A suspension of enzymatically dispersed nasal epithelium seeded on collagen foil (5 x 10(5) cells) served as control. Cell growth and ciliary beat were monitored through an inverted microscope with Hoffman's modulation contrast and video set-up. Histological examination was performed after 4, 6 and 8 weeks.
In the tissue cultures, the collagen foil was initially covered with fibroblasts growing from the mucosa specimen before epithelial cells spread out. The epithelial layer showed mostly ciliated cells which developed metachronous ciliary beat after 4 weeks in vitro. Ciliary activity was observed until the end of the experiments in 8 weeks. New cells on the suspension cultures were mesenchymal and did not exhibit any ciliary activity.
Mucosa specimens seem to be more appropriate for tissue engineering of respiratory epithelium than cell suspensions from nasal epithelium. Collagen foil as tissue scaffold initiates epithelial-mesenchymal interaction and may play an important role in epithelial differentiation of new respiratory epithelium.
生物人工气管假体的临床应用仍被视为一种实验性概念,因为在假体表面恢复功能性呼吸上皮仍然无法实现。组织工程作为一门相对较新的生物技术学科,可能会提供在体外扩增分化的呼吸上皮的新方法。在本研究中,我们比较了两种不同的细胞和组织培养程序,用于在市售胶原箔上培养人鼻黏膜。
采集人鼻黏膜标本(n = 6,4×4 cm),置于胶原箔上,作为组织培养物孵育4、6和8周。将酶分散的鼻上皮细胞悬液接种在胶原箔上(5×10⁵个细胞)作为对照。通过带有霍夫曼调制对比度的倒置显微镜和视频装置监测细胞生长和纤毛摆动。在4、6和8周后进行组织学检查。
在组织培养中,胶原箔最初被从黏膜标本中生长出来的成纤维细胞覆盖,然后上皮细胞才扩散开来。上皮层主要显示为纤毛细胞,在体外培养4周后出现同步纤毛摆动。在8周的实验结束前均观察到纤毛活动。悬浮培养中的新细胞是间充质细胞,未表现出任何纤毛活动。
黏膜标本似乎比鼻上皮细胞悬液更适合用于呼吸上皮的组织工程。胶原箔作为组织支架可引发上皮-间充质相互作用,并可能在新呼吸上皮的上皮分化中发挥重要作用。