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不同表面处理方法对聚乙醇酸支架物理、化学和生物学性能的影响。

The effect of different surface treatment methods on the physical, chemical and biological performances of a PGA scaffold.

作者信息

Song Yimin, Ren Minghua, Wu Yadong, Li Siyu, Song Chun, Wang Fang, Huang Yudong

机构信息

Department of Health Medicine, Peking Union Medical College Hospital Beijing 100730 China.

Surgery Department, the Key Laboratory of Cell Transplantation of Ministry of Health of the First Affiliated Hospital of Harbin Medical University Harbin 150001 China.

出版信息

RSC Adv. 2019 Jun 28;9(35):20174-20184. doi: 10.1039/c9ra02100k. eCollection 2019 Jun 25.

Abstract

In order to improve the adhesion between a PGA scaffold and islet cells, it is necessary to find a suitable method to modify the scaffold. In this study, the PGA scaffold surface was modified by plasma, polylysine coating and plasma combined with polylysine coating (P-P-PGA). The surface adhesion of the modified PGA scaffold was examined, and the stretchability and infiltration of the PGA scaffold were also tested. Then, the PGA scaffold treated under the optimal treatment conditions was selected to co-culture with rat islet cells, and the survival activity of the rat islet cells on the untreated PGA scaffold and the P-P-PGA scaffold was examined the MTT method. Rhodamine staining and DAPI staining were used to detect the number of islet cells adhered to four groups of scaffolds at different culture time points. The PGA-islet graft in the leg muscle of rats was stained with HE to perform the PGA-islet graft pathological examination. The experimental results showed that when the plasma treatment power was 240 W, the processing time was 4 min; the concentration of the polylysine coating solution was 2 mg ml, the tensile strength of the PGA scaffold was 320.45 MPa and the amount of infiltration of the PGA scaffold by the serum medium presented the maximum value: 3.17 g g. The MTT survival activity test results showed that after 3 d of culture, the survival activity of the islet cells of the treated PGA scaffold culture group (2.02 ± 0.13) was significantly different from the survival activity of the islet cells of the untreated PGA scaffold culture group (1.93 ± 0.10). The survival activities of the islet cells in the experimental groups (1.60 ± 0.13, 1.40 ± 0.12) were still higher than those of the control groups (0.96 ± 013, 0.69 ± 0.09) at 15 and 21 d. The results of the rhodamine and DAPI staining showed that with the increase in culture time, the number of the adherent cells in each group increased, and the number of the adherent islet cells in the experimental group was higher than that in the untreated group. The HE staining results showed that the islet cells on the P-P-PGA scaffold were more than those on the untreated PGA scaffold. After modification of the PGA scaffold, the adhesion of the islet cells improved, which was conducive to the growth of islet cells. These results confirmed that the plasma combined with polylysine coating treatment could enhance the adhesion of the PGA scaffold surface, so that the scaffold and the islet cells exhibited better adhesion and biocompatibility, and the modified PGA scaffold (P-P-PGA) could be used as a promising islet cell scaffold.

摘要

为了提高聚乙醇酸(PGA)支架与胰岛细胞之间的黏附力,有必要找到一种合适的方法对支架进行改性。在本研究中,采用等离子体、聚赖氨酸包被以及等离子体与聚赖氨酸联合包被(P-P-PGA)的方法对PGA支架表面进行改性。检测改性PGA支架的表面黏附力,并测试PGA支架的拉伸性和渗透性。然后,选择在最佳处理条件下处理的PGA支架与大鼠胰岛细胞共培养,采用MTT法检测大鼠胰岛细胞在未处理的PGA支架和P-P-PGA支架上的存活活性。利用罗丹明染色和DAPI染色检测不同培养时间点四组支架上黏附的胰岛细胞数量。对大鼠腿部肌肉中的PGA-胰岛移植物进行苏木精-伊红(HE)染色,以进行PGA-胰岛移植物的病理检查。实验结果表明,当等离子体处理功率为240 W、处理时间为4 min;聚赖氨酸包被溶液浓度为2 mg/ml时,PGA支架的拉伸强度为320.45 MPa,血清培养基对PGA支架的渗透量呈现最大值:3.17 g/g。MTT存活活性测试结果显示,培养3 d后,处理后的PGA支架培养组胰岛细胞的存活活性(2.02±0.13)与未处理的PGA支架培养组胰岛细胞的存活活性(1.93±0.10)有显著差异。在第15天和第21天,实验组胰岛细胞的存活活性(1.60±0.13,1.40±0.12)仍高于对照组(0.96±0.13,0.69±0.09)。罗丹明和DAPI染色结果表明,随着培养时间的增加,各组黏附细胞数量增加,实验组黏附的胰岛细胞数量高于未处理组。HE染色结果显示,P-P-PGA支架上的胰岛细胞比未处理的PGA支架上的多。PGA支架改性后,胰岛细胞的黏附力提高,有利于胰岛细胞的生长。这些结果证实,等离子体与聚赖氨酸联合包被处理可增强PGA支架表面的黏附力,使支架与胰岛细胞表现出更好的黏附性和生物相容性,改性后的PGA支架(P-P-PGA)可作为一种有前景的胰岛细胞支架。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/9065566/c416db9136fa/c9ra02100k-f1.jpg

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