Repurposing amniotic membrane as a native scaffold for cancer cell invasion studies

Abstract

Modeling cancer cell invasion requires physiologically relevant systems, yet traditional 2D/3D assays and animal models fail to capture the biochemical and mechanical complexity of the human extracellular matrix (ECM). The human amniotic membrane (AM) is a clinically approved, abundant, and immunologically privileged tissue with a rich ECM composition and favorable mechanical properties. Despite its extensive use in regenerative medicine, its potential as a cancer invasion scaffold remains underexplored. We propose repurposing decellularized AM (dAM) as a human-derived ECM platform to study tumor invasion. dAM retains structural proteins, growth factor reservoirs, and stiffness gradients that influence epithelial-to-mesenchymal transition (EMT) and invasion pathways. Compared with conventional matrices, it offers improved biochemical fidelity and compatibility with patient-derived organoids. Key challenges, including donor variability, decellularization optimization, and reproducibility, are also addressed. dAM provides a non-invasive, scalable, and physiologically relevant tool for cancer invasion assays, drug screening, and patient-specific models. Its integration into oncology research may enhance translational relevance and accelerate personalized medicine.

Keywords

amniotic membrane / cancer invasion / EMT

References

[1] Brabletz T, Kalluri R, Nieto MA, Weinberg RA. EMT in cancer. Nat Rev Cancer. 2018; 18: 128-34.

[2] Tuveson D, Clevers H. Cancer modeling meets human organoid technology. Science. 2019; 364: 952-55.

[3] Aisenbrey EA, Murphy WL. Synthetic alternatives to Matrigel. Nat Rev Mater. 2020; 5: 539-51.

[4] Sanders FWB, Huang J, Barrio JLAD, Hamada S, McAlinden C. Amniotic membrane transplantation: structural and biological properties, tissue preparation, application and clinical indications. Eye (Lond). 2024; 38: 668-79.

[5] Milan PB, Amini N, Joghataei MT, Ebrahimi L, Amoupour M, Sarveazad A, et al. Decellularized human amniotic membrane: From animal models to clinical trials. Methods. 2020; 171: 11-9.

[6] Ahmed K, Tauseef H, Ainuddin JA, Zafar M, Khan I, Salim A, et al. Assessment of the proteome profile of decellularized human amniotic membrane and its biocompatibility with umbilical cord-derived mesenchymal stem cells. J Biomed Mater Res A. 2024; 112: 1041-56.

[7] Abou-Shanab AM, Gaser OA, Soliman MW, Oraby A, Salah RA, Gabr M, et al. Human amniotic membrane scaffold enhances adipose mesenchymal stromal cell mitochondrial bioenergetics promoting their regenerative capacities. Mol Cell Biochem. 2025; 480: 2611-32.

[8] Guo M, Wu K, Yang M, Yin Y, Li Z, Wang G, et al. Bovine amniotic membrane with antioxidant and anti-inflammatory properties for the repair of alkali-burned corneas. Acta Biomater. 2025; 201: 198-211.

[9] Jafari A, Mirzaei Y, Mer AH, Rezaei-Tavirani M, Jafari Z, Niknejad H. Comparison of the effects of preservation methods on structural, biological, and mechanical properties of the human amniotic membrane for medical applications. Cell Tissue Bank. 2024; 25: 305-23.

[10] Esmaeili Z, Nokhbedehghan Z, Alizadeh S, majidi J, Chahsetareh H, Daryabari SH, et al. Biomimetic amniotic/silicone-based bilayer membrane for corneal tissue engineering. Mater Des. 2024; 237: 112614.

[11] Ganjibakhsh M, Mehraein F, Koruji M, Aflatoonian R, Farzaneh P. Three-dimensional decellularized amnion membrane scaffold as a novel tool for cancer research; cell behavior, drug resistance and cancer stem cell content. Mater Sci Eng C Mater Biol Appl. 2019; 100: 330-40.

[12] Abou-Shanab AM, Gaser OA, Salah RA, El-Badri N. Application of the Human Amniotic Membrane as an Adjuvant Therapy for the Treatment of Hepatocellular Carcinoma. Adv Exp Med Biol. 2024; 1470: 129-46.

[13] Etchebarne M, Fricain J, Kerdjoudj H, Pietro RD, Wolbank S, Gindraux F, et al. Use of Amniotic Membrane and Its Derived Products for Bone Regeneration: A Systematic Review. Front Bioeng Biotechnol. 2021; 9: 661332.

[14] Wells HC, Sizeland KH, Kirby N, Haverkamp RG. Structure and Strength of Bovine and Equine Amniotic Membrane. Biology (Basel). 2022; 11: 1096.

[15] Peng C, Zhang Q, Yang Q, Zhu Q. Strain and stress variations in the human amniotic membrane and fresh corpse autologous sciatic nerve anastomosis in a model of sciatic nerve injury. Neural Regen Res. 2012; 7: 1779-85.

[16] Mao Y, John N, Protzman NM, Long D, Sivalenka R, Azimi S, et al. A tri-layer decellularized, dehydrated human amniotic membrane scaffold supports the cellular functions of human tenocytes in vitro. J Mater Sci Mater Med. 2023; 34: 37.

[17] Tanaka K, Nagayama T, Katayama T, Koizumi N. Tensile properties of amniotic membrane. High Perform Struct Mater. 2010; 112: 197-206.

[18] Sarrigiannidis SO, Rey JM, Dobre O, González-García C, Dalby MJ, Salmeron-Sanchez M. A tough act to follow: collagen hydrogel modifications to improve mechanical and growth factor loading capabilities. Mater Today Bio. 2021; 10: 100098.

[19] Niknejad H, Peirovi H, Jorjani M, Ahmadiani A, Ghanavi J, Seifalian AM. Properties of the amniotic membrane for potential use in tissue engineering. Eur Cell Mater. 2008; 15: 88-99.

[20] Sharma R, Nappi V, Empeslidis T. The developments in amniotic membrane transplantation in glaucoma and vitreoretinal procedures. Int Ophthalmol. 2023; 43: 1771-83.

[21] Hu Z, Luo Y, Ni R, Hu Y, Yang F, Du T, et al. Biological importance of human amniotic membrane in tissue engineering and regenerative medicine. Mater Today Bio. 2023; 22: 100790.

[22] Abou-Shanab AM, Fytory M, Shouman S, Atta D, Khairy A, Salah RA, et al. Amniotic membrane-encapsulated chitosan-lecithin nanoparticles promote the regenerative potential of mesenchymal stromal cells and fibroblasts. Nanoscale Adv. 2025; 7: 5042-57.

[23] Abou-Shanab AM, Shouman S, Hussein AE, Gaser OA, Magdy S, Ashraf E, et al. Amniotic membrane promotes doxorubicin potency by suppressing SH-SY5Y neuroblastoma cell angiogenesis. BMC Cancer. 2025; 25: 1021.

[24] Abou-Shanab AM, Gaser OA, Galal N, Mohamed A, Atta D, Kamar SS, et al. PHD-2/HIF-1α axis mediates doxorubicin-induced angiogenesis in SH-SY5Y neuroblastoma microenvironment: a potential survival mechanism. Sci Rep. 2025; 15: 7487.

[25] Baig IF, Le NT, Al-Mohtaseb Z. Amniotic membrane transplantation: an updated clinical review for the ophthalmologist. Ann Eye Sci. 2023; 8: 5.

[26] Hjazi A. A collagen-based amniotic membrane scaffold combined with photobiomodulation accelerates wound repair in diabetic rats through modulation of inflammation and tissue regeneration. Tissue Cell. 2025; 97: 103063.

[27] Elkhenany H, Abou-Shanab AM, Magdy S, Kamar SS, Salah RA, El Badri N. Comprehensive evaluation of ethanol-preserved amniotic extracts: Exploring antioxidant properties, proliferation enhancement, protective efficacy and regeneration potential in wound healing. J Drug Delivery Sci Technol. 2024; 100: 106062.

[28] Chen H, Song G, Xu T, Meng C, Zhang Y, Xin T, et al. Biomaterial Scaffolds for Periodontal Tissue Engineering. J Funct Biomater. 2024; 15: 233.

[29] Guimarães M, Calheiro GAT, Sant’Anna LB. Analysis of the Protective Potential of the Amniotic Membrane in an In Vitro Experimental Model of Demyelination in Mouse Brain Organotypic Slices . ACS Omega. 2025; 10: 33162-77.

[30] Atta D, Abou-Shanab AM, Kamar SS, Soliman MW, Magdy S, El-Badri N. Amniotic Membrane-Derived Extracellular Matrix for Developing a Cost-Effective Xenofree Hepatocellular Carcinoma Organoid Model. J Biomed Mater Res A. 2025; 113: e37882.

[31] Marshall J. Transwell^® Invasion Assays . In: Wells CM, Parsons M, editors. Cell Migration. Humana Totowa: Springer. 2011. pp. 97-110.

[32] Hira VV, Breznik B, Van Noorden CJ, Lah T, Molenaar RJ. 2D and 3D in vitro assays to quantify the invasive behavior of glioblastoma stem cells in response to SDF-1α. Biotechniques. 2020; 69: 339-46.

[33] Elkin M, Vlodavsky I. Tail vein assay of cancer metastasis. Curr Protoc Cell Biol. 2001; Chapter 19: 19.2.1-19.2.7.