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Supportive properties of basement membrane layer of human amniotic membrane enable development of tissue engineering applications

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Abstract

Human amniotic membrane (HAM) has been widely used as a natural scaffold in tissue engineering due to many of its unique biological properties such as providing growth factors, cytokines and tissue inhibitors of metalloproteinases. This study aimed at finding the most suitable and supportive layer of HAM as a delivery system for autologous or allogeneic cell transplantation. Three different layers of HAM were examined including basement membrane, epithelial and stromal layers. In order to prepare the basement membrane, de-epithelialization was performed using 0.5 M NaOH and its efficiency was investigated by histological stainings, DNA quantification, biomechanical testing and electron microscopy. Adipose-derived stromal cells (ASCs) and a human immortalized keratinocyte cell line (HaCaT) were seeded on the three different layers of HAM and cultured for 3 weeks. The potential of the three different layers of HAM to support the attachment and viability of cells were then monitored by histology, electron microscopy and (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Moreover, mechanical strengths of the basement membrane were assessed before and after cell culture. The results indicated that the integrity of extra cellular matrix (ECM) components was preserved after de-epithelialization and resulted in producing an intact basement amniotic membrane (BAM). Moreover, all three layers of HAM could support the attachment and proliferation of cells with no visible cytotoxic effects. However, the growth and viability of both cell types on the BAM were significantly higher than the other two layers. We conclude that growth stimulating effectors of BAM and its increased mechanical strength after culturing of ASCs, besides lack of immunogenicity make it an ideal model for delivering allogeneic cells and tissue engineering applications.

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Abbreviations

AECs:

Amniotic epithelial cells

AM:

Amniotic membrane

AMCs:

Amniotic mesenchymal cells

ASCs:

Adipose-derived stromal cells

BM:

Basement membrane

DAPI:

4′,6-Diamidino-2-phenylindole

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

Dimethyl sulfoxide

EAM:

Epithelial amniotic membrane

ECM:

Extracellular matrix

EDTA:

Ethylenediaminetetraacetic acid

EGF:

Epidermal growth factor

FBS:

Fetal bovine serum

GAG:

Glycosaminoglycan

HaCaT:

Human immortalized keratinocyte cell line

HAM:

Human amniotic membrane

H–E:

Hematoxylin and eosin

KGF:

Keratinocyte growth factor

MSCs:

Mesenchymal stem cells

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

OD:

Optical density

PBS:

Phosphate buffered saline

PDGF:

Platelet-derived growth factor

SAM:

Stromal amniotic membrane

SEM:

Scanning electron microscope

TEM:

Transmission electron microscope

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Acknowledgements

This work was supported by Ferdowsi University of Mashhad (Grant Number 3.38091) and also ACECR-Khorasan Razavi Branch. We gratefully acknowledge the collaboration of Dr. Nasser Sanjar Moosavi in supplying liposuction materials and also Fatemeh Vakili for providing the HAMs. In addition, the authors gratefully acknowledge Fatemeh Naseri and  Mahmood Raeesolmohaddesin for their technical assistance.

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

    Sonia Iranpour, Nasser Mahdavi-Shahri & Maryam M. Matin

  2. Research Center for HIV/AIDS, HTLV and Viral Hepatitis, ACECR-Khorasan Razavi Branch, Mashhad, Iran

    Raheleh Miri

  3. Stem Cells and Regenerative Medicine Research Group, ACECR-Khorasan Razavi Branch, Mashhad, Iran

    Halimeh Hasanzadeh, Hamid Reza Bidkhori, Hojjat Naderi-Meshkin & Maryam M. Matin

  4. Dental Materials Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Ehsan Zahabi

  5. Novel Diagnostics and Therapeutics Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

    Maryam M. Matin

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  1. Sonia Iranpour
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  2. Nasser Mahdavi-Shahri
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Correspondence to Maryam M. Matin.

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Iranpour, S., Mahdavi-Shahri, N., Miri, R. et al. Supportive properties of basement membrane layer of human amniotic membrane enable development of tissue engineering applications. Cell Tissue Bank 19, 357–371 (2018). https://doi.org/10.1007/s10561-017-9680-z

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