纖維母細胞培養於不同生物薄膜所引發之類似老化之表徵

Abstract

前言：回顧過去組織工程的研究可知，不同生醫材料表面的化學和物理性質與細胞的生存、生長等細胞行為表現息息相關，但至今尚未有文獻指出，用於培養細胞之生醫材料對於細胞老化上的影響。因此，本研究的主要目標為觀察不同之高分子培養薄膜對於細胞老化的影響。 材料與方法：實驗選用具有聚乙烯莫耳比44的聚乙烯－乙烯醇共聚合物(EVAL44)、具有聚乙烯莫耳比為27的聚乙烯－乙烯醇共聚合物(EVAL27)、商業用組織培養盤(TCPS)三種常見的高分子薄膜作為實驗材料，並以繼代培養時是否更換鋪有不同材料之培養盤為條件，將實驗條件區分為兩組。利用真皮纖維母細胞作為實驗細胞，培養於不同的高分子培養薄膜上，建立細胞長期之生長曲線與 SA-β-galactosidase酵素活性等老化相關指標，最後進行細胞骨架染色與相關的基因分析。 結果：實驗結果顯示，在更換培養盤這一組實驗中，培養於三種高分子培養薄膜上的細胞在生長曲線、SA-β-galactosidase酵素活性、細胞型態變化、和老化相關基因的表現，產生了明顯的差異性，且在細胞骨架染色的部分，也可以發現細胞上的Focal adhesion與細胞骨架均有所不同；但在不更換培養盤這組實驗中並沒有發現相同的現象，培養於三種高分子培養薄膜上的細胞在老化表徵上並沒有產生差異性。 結論：不同高分子培養薄膜所提供的培養環境，明顯的影響細胞產生類似於老化之行為，而細胞外基質於材料上的型態與堆積之數量與高分子培養薄膜的表面親疏水性，或許會是影響細胞於培養材料上老化的主要原因。

Introduction: It has been known that different chemical and physical surface properties of biomembranes controlled the cell behavior performance, such as cell survival, and growth. However, there is no report which is focused on the effects of biomembranes to cellular senescence. Thus, the objective of this study was to investigate the influence of different biomembranes on cellular senescence. Materials and methods: Three kinds of common polymer materials, Poly(vinyl alcohol-co-ethylene) with 44mol% ethylene (EVAL44), Poly(vinyl alcohol-co- ethylene) with 27mol% ethylene (EVAL 27) , and tissue culture polystyrene (TCPS) , were selected as test materials, and there were two groups under the condition of culture plate replacement or not. Primary skin dermal fibroblasts were cultured on these membranes to establish senescence correlation index, such as growth curves and SA-β-galactosidase activity. Finally, the cell skeleton and senescence associated gene analysis were carried out. Results: In this study, the group with culture plate replacement demonstrated that the cells on three biomembranes have obvious differences in the growth curve, SA-β-galactosidase activity, cell morphology, senescence associated genes, and cell skeleton. According to the results of cell skeleton staining, focal adhesion and cell skeleton in three kinds of biomembranes have distinct performances. The same phenomenon was not observed in cells that grew in another group with no culture plate replacement. Conclusions: This study showed that the environment that provided by different polymer biomembranes obviously affected the behavior of senescence-like phenotype. The formation or the amount of extracellular matrix piling up on the membrane and the surface hydrophobicity of biomembrane might be the main influence factors.