Effects of Attachment and Proliferation of Retinal Pigment Epithelial Cells on Silk/PLGA Film

Abstract

초록: 망막 조직공학을 위한 생체 재료는 기계적 안전성, 생체적합성, 낮은 분해속도 등을 포함하여 in vivo에서 잠 재적인 유용성을 위한 몇 가지 중요한 특징이 입증되어야 한다. 실크 필름 생체재료는 이러한 기능적인 요구에 맞게 디자인되었다. 0, 10, 20, 40, 및 80 wt%의 실크가 함유된 천연/합성물질과 하이브리드화된 silk/PLGA 필름을 용매 증발법으로 제조하였다. 1, 2, 및 3일 후에 부착된 세포 수를 확인하기 위해 MTT 분석을 하였고 SEM을 통해 필름 에 부착된 세포 모폴로지를 확인하였다. 또한, mRNA 발현정도를 알아보기 위해 retinal pigment epithelium(RPE) 세 포의 프라이머인 RPE65를 사용하여 RT-PCR을 실시하였고 RPE 세포의 특정 단백질인 cytokeratin의 발현을 확 인하고 세포의 증식을 비교하기 위해 면역화학염색을 실시하였다. 본 실험을 통해 실크/PLGA 필름에서 20∼40 wt% 실크를 함유한 경우에 RPE 세포의 부착과 증식에 가장 좋은 영향을 미치는 것을 확인하였다.Abstract: Biomaterials for retinal tissue engineering must demonstrate several critical features for potential utility, including mechanical integrity, biocompatibility, and slow biodegradation. Silk film biomaterials were designed and characterized to meet these functional requirements. We prepared natural/synthetic hybrid silk/PLGA films using 0, 10, 20, 40, and 80 wt% of silk by a solvent evaporation method. MTT assay was used to confirm the number of cells attached on film at 1, 2, and 3 days, respectively. The morphology of cellular adhesion on films was also confirmed by scanning electron microscope (SEM). RT-PCR was conducted to confirm mRNA expression of retinal pigment epithelium(RPE) using RPE65 as a RPEs marker and the expression of cytokeratin were determined by immunofluorescence staining. We confirmed that the silk/PLGA film of 20∼40 wt% silk was superior for the adhesion and proliferation of RPEs.

A study of a three-dimensional PLGA sponge containing natural polymers co-cultured with endothelial and mesenchymal stem cells as a tissue engineering scaffold

“…The PLGA, PLGA/DBP, PLGA/silk, PLGA/SIS scaffolds were prepared using the solvent casting/salt leaching technique [42]. Briefly, 1 g of PLGA was dissolved in 4 ml of methylene chloride (J. T. Baker, USA), and then 9 g of sodium chloride (180 ~ 250 µm) was added to the solution to create the porous structure in the scaffolds with or without various natural polymer powders (20 wt% of PLGA) because 20% concentration of natural polymers was suitable for attachment, proliferation, differentiation and inflammatory response of cells in our previous studies [23,43,44]. Scaffolds were pressed at room temperature for 24 h after inserting the solution in a silicon mold which has a disc shape (1 cm × 0.5 cm width, 0.3 cm thickness).…”

Adhesion and Proliferation Behavior of Retinal Pigment Epithelial Cells on Hesperidin/PLGA Films

“…[13][14][15] 그러나 생체활성물질이 부족하고 소수성 성질로 인한 세포 부착 및 주변조직과의 융합에 어려움이 있 어 세포의 활성이 낮은 단점이 있다. 16 이러한 PLGA의 단점 을 보완하고자 사용한 헤스페리딘(3',5,7-trihydroxy-4-methoxyflavanone-7-rhamno-glucoside)은 감귤류에서 풍부하게 발견 되는 플라바논 글리코사이드이다( Figure 1). 헤스페리딘은 항 산화 효과, 항염증 효과, 항-고콜레스테롤혈증 활성, 항-고혈 압성 활성, 이뇨특성을 갖고 있다.…”