Evaluation of Fibroblast Viability Seeded on Acellular Human Amniotic Membrane.

Hamideh Moravvej,Maryam Kabir-Salmani,Mojtaba Memariani,Hamed Memariani,Alireza Shoae-Hassani,Fahimeh Abdollahimajd,Maciej Gagat

doi:10.1155/2021/5597758

Abstract

Background Investigating the viability and proliferative rates of fibroblast cells on human amniotic membrane (HAM) as a scaffold will be an important subject for further research. The aim of this study was to assess the fibroblast viability seeded on acellular HAM, since foreskin neonatal allogenic fibroblasts seeded on HAM accelerate the wound healing process. Methods Fibroblasts were retrieved from the foreskin of a genetically healthy male infant, and we exploited AM of healthy term neonates to prepare the amniotic scaffold for fibroblast transfer. After cell culture, preparation of acellular HAM, and seeding of cells on HAM based on the protocol, different methods including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 4′,6-Diamidino-2-phenylindole dihydrochloride (DAPI), and propidium iodide (PI) staining were employed for assessment of fibroblast viability on HAM. Results Based on the results obtained from the DAPI and PI staining, the percentage of viable cells in the former staining was clearly higher than that of the dead cells in the latter one. The results of DAPI and PI staining were in accordance with the findings of MTT assay, confirming that fibroblasts were viable and even proliferate on HAM. Conclusion Our findings showed the viability of fibroblasts seeded on the acellular HAM using MTT assay, DAPI, and PI staining; however, this study had some limitations. It would be an interesting subject for future research to compare the viability and proliferation rate of fibroblasts seeded on both cellular and acellular HAM.

Highlights

As one of the major public health problems, chronic wounds lead to increased morbidity, disability, and risk of mortality, imposing a considerable financial burden on both patients and healthcare system [1, 2]
MTT assay indicated a significant increase in viability when fibroblast cells were cultured on the amniotic membrane (P < 0:05) compared to control (Figure 2)
There was a statistically significant increase in cell viability in the sample group compared to the control groups (P < 0:05; Figure 2)

Summary

Introduction

As one of the major public health problems, chronic wounds lead to increased morbidity, disability, and risk of mortality, imposing a considerable financial burden on both patients and healthcare system [1, 2]. Preparation of acellular HAM, and seeding of cells on HAM based on the protocol, different methods including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 4′,6Diamidino-2-phenylindole dihydrochloride (DAPI), and propidium iodide (PI) staining were employed for assessment of fibroblast viability on HAM. The results of DAPI and PI staining were in accordance with the findings of MTT assay, confirming that fibroblasts were viable and even proliferate on HAM. Our findings showed the viability of fibroblasts seeded on the acellular HAM using MTT assay, DAPI, and PI staining; this study had some limitations.

Objectives

Methods

Results

Discussion

Conclusion