title: The effect of cryopreservation on anti-cancer activity of human amniotic membrane.
abstract: Human amniotic membrane (AM) is an appropriate candidate for treatment of cancer due to special properties, such as inhibition of angiogenesis and secretion of pro-apoptotic factors. This research was designed to evaluate the impact of cryopreservation on cancer cell death induction and anti-angiogenic properties of the AM. Cancer cells were treated with fresh and cryopreserved amniotic condition medium during 24 h and cancer cell viability was determined by MTT assay. To evaluate angiogenesis, the rat aorta ring assay was performed for both fresh and cryopreserved AM within 7 days. In addition, four anti-angiogenic factors Tissue Inhibitor of Matrix Metalloproteinase-1 and 2 (TIMP-1 and TIMP-2), Thrombospondin, and Endostatin were measured by ELISA assay before and after cryopreservation. The results showed that the viability of cultured cancer cells dose-dependently decreased after treatment with condition medium of fresh and cryopreserved tissue and no significant difference was observed between the fresh and cryopreserved AM. The results revealed that the amniotic epithelial stem cells inhibit the penetration of fibroblast-like cells and angiogenesis. Moreover, the penetration of fibroblast-like cells in both epithelial and mesenchymal sides of fresh and cryopreserved AM was observed after removing of epithelial cells. The cryopreservation procedure significantly decreased anti-angiogenic factors TIMP-1, TIMP-2, Thrombospondin, and Endostatin which shows that angio-modulatory property is not fully dependent on proteomic and metabolomic profiles of the AM. These promising results demonstrate that cancer cell death induction and anti-angiogenic properties of the AM were maintained within cryopreservation; a procedure which can circumvent limitations of the fresh AM.
authors.affiliation: Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Biomaterials, Faculty of Biomedical Engineering, Amirkabir University of Technology, Tehran, Iran.
authors.forename: Khashayar
authors.initials: K
authors.lastname: Modaresifar
authors.affiliation: Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Biomaterials, Faculty of Biomedical Engineering, Amirkabir University of Technology, Tehran, Iran.
authors.forename: Sara
authors.initials: S
authors.lastname: Azizian
authors.affiliation: Nanomedicine and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Zolghadr
authors.affiliation: Skin Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Hamideh
authors.initials: H
authors.lastname: Moravvej
authors.affiliation: Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Abolhassan
authors.initials: A
authors.lastname: Ahmadiani
authors.affiliation: Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Nanomedicine and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address: niknejad@sbmu.ac.ir.
authors.forename: Hassan
authors.initials: H
authors.lastname: Niknejad
mesh_terms: D000650:Amnion / Q000166:cytology / Q000502:physiology*; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002452:Cell Count; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D043169:Endostatins / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D009369:Neoplasms / Q000473:pathology*; D009389:Neovascularization, Pathologic / Q000473:pathology*; D051381:Rats; D013234:Stem Cells / Q000502:physiology; D019699:Thrombospondins / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D043169:Endostatins; C560822:TIMP1 protein, human; C560823:TIMP2 protein, human; D019699:Thrombospondins; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1
keywords: Amniotic membrane; Angiogenesis; Apoptosis; Cancer; Cryopreservation; Epithelial cell