Fabrication and evaluation of collagen scaffolds for endometrial cells growth and proliferation in order to endometrial tissue regeneration

Abstract

Introduction: Due to the fact that endometrium related diseases are one of the basic and important factors of female infertility, researchers have closely scrutinized endometrial tissue regeneration. Endometrium as the innermost layer of the uterus plays a significant role in women's fertility and reproductive health.Aims: Here, we explain the stages of design and development of the biocompatible COL/PVA nanofibrous scaffolds in order to investigate the effects of HUC-MSCs loaded scaffolds in endometrium repair and regeneration.Methods: In this study, by optimizing the percentage ratio of COL to PVA, the need for cross linking process to maintain the nanofibers stability in aqueous environments had been eliminated. Chemical structure of synthesized scaffolds was evaluated by FT-IR. In addition, other physicochemical and biological aspects of the fabricated scaffolds, including nanofiber diameter, in vitro degradation, swelling behavior, mechanical properties, morphologies, and biocompatibility were surveyed. In this research, the indirect co-culture method was used with the aim of simulating the relationship between different types of cells in natural tissues to investigate the effect of HUC-MSCs loaded COL/PVA nanofiber scaffolds on the HESCs proliferation.Results: Physiochemical assessments showed that scaffolds with well-developed porous morphology and stable structure was obtained. None of synthesized scaffolds had no toxicity on HUC-MSCs. SEM images showed proper cell adhesion and distribution of HUC-MSCs throughout the nonafibrous scaffolds. The effects of HUC-MSCs loaded uncross-linked COL/PVA nanofibrous scaffolds on HESCs proliferation via DAPI/Ki67 Immuno-fluorescence staining, and cell cycle analysis was investigated. According to obtained results, 3 days after the start of co-culture HUC-MSCs loaded COL/PVA nanofiber scaffolds with HESCs amount of KI67 protein expression in the test group compared to the control group increased 39/7%. In addition, HESCs cell cycle studies showed that, the cell population in the G2+M phases in the test group compared to the control group reached from 7.92% to 15.95% and had a significant increase.Conclusion: The results demonstrate that biocompatible and biodegradable COL/PVA nanofibrous scaffolds hold potential for endometrium regeneration applications.