Study of electrospun polycaprolacton/gelatin nanofiber scaffold for uterine tissue engineering

Introduction In this study, a novel polycaprolactone/gelatin (PCL/G) scaffold with improved mechanical properties and controllable porous structure was prepared through electro spinning method. In this research, the biocompatibility was evaluated, using endometrial cells. MTT results indicated that the growth of endometrial cells developed in the PCL/G scaffold. Although further investigations, such as the extracellular matrix production and the functional gene expression, are in progress to confirm the MTT results. Our data suggests that the PCL/G scaffold is a new suitable scaffold material for tissue engineering. Objectives The purpose of this project is to eliminate of problems similar to damaged uterus, rennet uterus (surrogacy), lack of womb for womens using a scaffold like uterine tissue. Methods Polycaprolactone (PCL) and gelatin (G) were purchased from Sigma-Aldrich® and trifluoroacetic acid (C2HF3O2) from Merck® companies. PCL/G copolymer was synthesized with volume ratios of 10:0, 0:10, 9:1, 7:3 and 5:5 (PCL:G). Fibers were electrospun on the aluminium foil. The resulting nanofibers were used for characterization and cell culture was tested on days 1, 3 and 7. Cell interaction and growth of endometrial cells on PCL/G fibers were investigated. Endometrial cell viability was studied by 3-[4,5-dimethylthiazol- 2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay. With an informed written consent, endometrial cells were extracted from normal human uterine tissue samples. Endometrial cells were cultured in standard condition with DMEM. Surface markers (CD90+, CD105+, CD146+, and CD45) distinguished them from mesenchymal stem cells (MSCs). After sterilization under UV light for 20 minutes, the electrospun fibers were placed in 24-well plates and seeded with endometrial cells at a density of 10,000 cells per well. The cells were incubated in a humidified standard incubator at 37 °C with 5% CO2. Cells were harvested at days 1, 3, and 7 for growth and phenotypical evaluation and MTT test. By triplicate assay the activity and survival of the cells on nanofibers were calculated as a percentage of samples to controls.Results The random andaligned scaffolds were developed by the electrospinning technique, and their morphological microstructures were investigated by SEM. SEM micrographs of the scaffolds were obtained before seeding of the cells. The endometrial cells on all four types of scaffolds were found to spread well and the cells were grown on the scaffold. Also, significant increase in cell growth and replication on the scaffold compared to control samples indicates the lack of toxicity of the wizard for the scaffold adaptation. Conclusion These findings could be used in tissue engineering as the first step for uterine tissue engineering. We planned to evaluate a fertilized egg’s reaction on this novel scaffold.

کلیدواژه ها:

Endometrium ، Nano fiber ، Polycaprolactone/Gelatin ، Scaffold ، Tissue engineering