Evaluation of Electrospun Polycaprolactone/Gelatin/ Polydimethylsiloxane Nanofiber Scaffold for Endometrial Cell Growthand Proliferation as Part of Uterine Tissue Engineering

Background and Aim: In this study a PCL- G- PDMS copolymer wasfurther engineered into porous fiber scaffolds by electrospinning. Thebiocompatibility was evaluated, using fibroblasts and endometrialcells and assessing cell metabolic activity by MTT assay. the aim of thisstudy was to develop and evaluate electrospun nano-fibrous scaffoldsto support the growth of endometrial cells. The aim of this project isto solve the problems similar to hysterectomized uterus, rennet uterus(surrogacy), and the lack or defective womb using an engineered uterinetissue.Methods: PCL- G- PDMS segments with different ratios weresynthesized. Cell interaction and growth of endometrial cells andendometrial cell viability were investigated by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay on PCL- G- PDMSfibers. With an informed written consent, endometrial cells wereextracted from normal human uterine tissue samples. The surfacemarkers (CD90+, CD105+, CD146+, and CD45)were distinguishedthem from mesenchymal stem cells (MSCs). After sterilization under UVlight for 20 minutes, the electrospun fibers were placed as a disk at thebottom of every well in a 24-well cell culture plate and were seededwith endometrial cells at a density of 10,000 cells per well. Cells wereharvested at days 1, 3, and 7 for growth and phenotypical evaluationand MTT test. By triplicate assay, the activity and survival of the cells onnanofibers were calculated as a percentage of samples to controls.Results: The electrospinning technique has been used to tailormicrostructures including fiber diameter and pore size of nano-fibrousscaffolds for tissue engineering applications. Electrospinning allows forthe generation of fibers with nanoscale diameters. Small fiber diametercombined with the high specific surface area, together with porousstructures of nano-fibrous scaffolds, are essential for tissue engineeringapplications. There are several parameters that can be manipulated totailor fiber diameter and pore size. These factors affect cell adhesion onthe scaffold. So the random, aligned scaffolds were developed by theelectrospinning technique, and their morphological microstructures wereinvestigated by SEM. SEM micrographs of the scaffolds were obtainedbefore seeding of the cells. The endometrial cells on all six types ofscaffolds were found to attach and spread well. There is a difference incell growth due to differences in fiber diameter and porosity per percent.Also, a significant increase in cell growth and replication on the scaffoldcompared to control samples indicates the lack of toxicity of the wizardfor the scaffold adaptation.Conclusion: In this study, we synthesized a series of novel biodegradablescaffolds comprising different ratios of PCL- G- PDMS. These PCL- GPDMScopolymers were further engineered via electrospinning. The PCLG-PDMS fibers showed decent mechanical properties and hold greatpotential for various biomedical applications. The advanced mechanicalproperties and biocompatibility of these PCL- G- PDMS fibers wouldallow them to become promising candidates for tissue engineering.Evaluation of endometrial cells on this scaffold could be the first stepfor uterine tissue engineering. We planned to evaluate a fertilized egg’sreaction to this novel scaffold.

کلیدواژه ها:

Endometrium ، Nano-fiber ، Polycaprolactone ، Gelatin ، Polydimethylsiloxane ، Tissue engineering