不同生物材料對內皮細胞生長之研究

Growth Endothelial Cell on GRGD Modified PU and Chitosan Surfaces

王欣雅(Hsin-Ya Wang)

指導教授 ： 鍾次文 ； 蔡正倫

中原大學/工學院/醫學工程研究所/碩士(2002年)

https://doi.org/10.6840/cycu200200537

摘要

本文研究內皮細胞（endothelial cell）在不同的生物材料上之貼附生長。主要探討的因素有兩種：（1）粗糙度對內皮細胞生長之影響、（2）材料上接枝Gly-Arg-Gly-Asp（GRGD）對內皮細胞生長之影響。
PU材料的粗糙度的形成，是以三種分子量（Mw = 1100、2000、5000）混合聚乙二醇分子（PEG）混合接枝在PU表面，使修飾的PU具較大粗糙度之表面（PU-PEGmix），經由原子力顯微鏡（AFM）之量測，PU-PEGmix平均粗糙度為56.80±35.09 nm（n=3）大於PU-PEG2000平均粗糙度為20.10±7.87 nm（n=3）及PU材料平均粗糙度為1.53±0.2 nm（n=3）；由細胞貼附生長實驗顯示，PU-PEGmix材料對細胞生長貼附量比PU-PEG2000高35％；且細胞活性測試（MTT test）也顯示，PU-PEGmix材料測試吸收值為0.028±0.003（n=6），其細胞活性比PU-PEG2000高25％。此外，將GRGD接枝到PU-PEG上形成PU-PEG-GRGD，經HPLC分析，GRGD接枝密度為4.2×10-8 mole/cm2；PU-PEG-GRGD細胞生長貼附量比PU-PEG高40％。且在細胞活性測試顯示，PU-PEG-GRGD材料材料測試吸收值為0.033±0.002（n=6），其細胞活性比PU-PEG高16％。
在幾丁聚醣方面，幾丁聚醣本研究經由光化學反應將GRGD接枝於材料上形成chitosan-GRGD，經FTIR測定G、R、D的存在，並由HPLC分析，知GRGD接枝密度為8.5×10-8 mole/cm2，另外，ESCA分析chitosan-GRGD其O/C值為0.46、N/C值為0.11皆比chitosan高。上述接枝chitosan-GRGD細胞貼附量比chitosan高52％；且在細胞活性測試顯示，chitosan-GRGD材料材料測試吸收值為0.04±0.01（n=6），其細胞活性比chitosan高65％。
結論，當生醫材料表面的粗糙度較大或具有GRGD peptides時，其內皮細胞貼附生長之情形較佳。

關鍵字

GRGD ； 幾丁聚醣 ； 聚胺基甲酸酯 ； 內皮細胞 ； 粗糙度

並列摘要

We investigated the adhesion and growth of human endothelial cells (HUVECs) on different biomaterials, polyurethane (PU) and chitosan. Two factors that might affect the growth HUVECs on the biomaterials have been studied. There are: (a) roughness of the surface, (b) Gly-Arg-Gly-Asp (GRGD) grafted on the surface.
To produce roughness of PU surface, we mixed equivalent molar of polyethylene glycol (PEG) of different molecular weight (Mw.) 1100, 2000 and 5000, and then grafted to PU surface (PU-PEGmix). The roughness defined by atomic force microscopic (AFM) measurements were 56.80±35.09 nm（n=3）, 20.10±7.87 nm and 1.53±0.2 nm for PU-PEGmix, PU-PEG2000 and PU surface, respectively. Regards to growth of HUVECs on those surfaces, the cell density were 35% higher than PU-PEG2000 surface, respectively. MTT test for cell viability also confirmed the results (e.g. The cell viability of HUVECs growth on PU-PEGmix was 25% higher than PU-PEG2000). In addition, PU-PEG grafted with GRGD to form PU-PEG-GRGD surface were also shown 40% higher growth density of HUVECs than PU-PEG without GRGD grafted although GRGD grafted density is only 4.2×10-8mole/cm2. MTT for HUVECs viability test for PU-PEG-GRGD was also show 16% (n=6) higher than without grafted.
The grafted density for GRGD in chitosan was 8.5×10-8mole/cm2. The HUVECs cell density for chitosan-GRGD surfaces are 52% higher than that for chitosan surface only. MTT test for cell viability of HUVECs growth on chitosan-GRGD surface was also 65％(n=6) higher than that of chitosan without GRGD grafted.
In conclusion, higher roughness of PU surface or GRGD peptide grafted on PU or chitosan surfaces could enhance HUVECs adhesion and growth.

並列關鍵字

polyurethane ； HUVEC ； roughness and GRGD ； chitosan

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