Assessment of the proteome profile of decellularized human amniotic membrane and its biocompatibility with umbilical cord-derived mesenchymal stem cells
Kainat Ahmed
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorHaadia Tauseef
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorJahan Ara Ainuddin
Dow University of Health Sciences, Karachi, Pakistan
Search for more papers by this authorMuneeza Zafar
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorIrfan Khan
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorAsmat Salim
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorMunazza Raza Mirza
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorCorresponding Author
Omair Anwar Mohiuddin
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Correspondence
Omair Anwar Mohiuddin, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan.
Email: [email protected]
Search for more papers by this authorKainat Ahmed
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorHaadia Tauseef
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorJahan Ara Ainuddin
Dow University of Health Sciences, Karachi, Pakistan
Search for more papers by this authorMuneeza Zafar
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorIrfan Khan
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorAsmat Salim
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorMunazza Raza Mirza
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Search for more papers by this authorCorresponding Author
Omair Anwar Mohiuddin
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan
Correspondence
Omair Anwar Mohiuddin, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan.
Email: [email protected]
Search for more papers by this authorKainat Ahmed and Haadia Tauseef contributed equally.
Abstract
Extracellular matrix-based bio-scaffolds are useful for tissue engineering as they retain the unique structural, mechanical, and physiological microenvironment of the tissue thus facilitating cellular attachment and matrix activities. However, considering its potential, a comprehensive understanding of the protein profile remains elusive. Herein, we evaluate the impact of decellularization on the human amniotic membrane (hAM) based on its proteome profile, physicochemical features, as well as the attachment, viability, and proliferation of umbilical cord-derived mesenchymal stem cells (hUC-MSC). Proteome profiles of decellularized hAM (D-hAM) were compared with hAM, and gene ontology (GO) enrichment analysis was performed. Proteomic data revealed that D-hAM retained a total of 249 proteins, predominantly comprised of extracellular matrix proteins including collagens (collagen I, collagen IV, collagen VI, collagen VII, and collagen XII), proteoglycans (biglycan, decorin, lumican, mimecan, and versican), glycoproteins (dermatopontin, fibrinogen, fibrillin, laminin, and vitronectin), and growth factors including transforming growth factor beta (TGF-β) and fibroblast growth factor (FGF) while eliminated most of the intracellular proteins. Scanning electron microscopy was used to analyze the epithelial and basal surfaces of D-hAM. The D-hAM displayed variability in fibril morphology and porosity as compared with hAM, showing loosely packed collagen fibers and prominent large pore areas on the basal side of D-hAM. Both sides of D-hAM supported the growth and proliferation of hUC-MSC. Comparative investigations, however, demonstrated that the basal side of D-hAM displayed higher hUC-MSC proliferation than the epithelial side. These findings highlight the importance of understanding the micro-environmental differences between the two sides of D-hAM while optimizing cell-based therapeutic applications.
CONFLICT OF INTEREST STATEMENT
The authors declare no conflict of interest.
Open Research
DATA AVAILABILITY STATEMENT
The data that supports the findings of this study are available in the supplementary material of this article.
Supporting Information
| Filename | Description |
|---|---|
| jbma37685-sup-0001-supplementary_data.xlsxExcel 2007 spreadsheet , 287.7 KB | Data S1. Supplementary Data. |
| jbma37685-sup-0002-Figure_S1.tifTIFF image, 699.1 KB | Figure S1. Human placental tissue processing. Amnion processing (A) human placental tissues were collected along with fetal membranes, amnion membrane separated from placenta and chorion, respectively, and washed to remove blood remnants. Preparation of biological scaffold. (B) hAM was decellularized using salt-based method, lyophilized and cut into 1 cm2 pieces. |
| jbma37685-sup-0003-Figure_S2.tifTIFF image, 356.3 KB | Figure S2. Human umbilical cord processing. Human umbilical cord were minced into 1 cm pieces, washed to remove blood and cultured into T-75 tissue culture flask. |
| jbma37685-sup-0004-Figure_S3.tifTIFF image, 774.5 KB | Figure S3. hUC-MSCs characterization. Representative illustrations of hUC-MSCs at (A) P0, P1, P2, and P3 passages (scale bar 100 μm). Proliferation of hUC-MSCs determined by population doubling time assay. (B) At day 1, 3, 5, and 7. Representative histograms (C) showing positive expression of MSCs markers that is, CD44 (PE-H), CD73 (APC-H), and CD90 (FITC-H) analyzed by flow cytometry. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1Dzobo K, Thomford NE, Senthebane DA, et al. Advances in regenerative medicine and tissue engineering: innovation and transformation of medicine. Stem Cells Int. 2018; 2018:2495848.
- 2Brovold M, Almeida JI, Pla-Palacín I, et al. Naturally-derived biomaterials for tissue engineering applications. Adv Exp Med Biol. 2018; 1077: 421-449.
- 3Alaribe FN, Manoto SL, Motaung SCJB. Scaffolds from biomaterials: advantages and limitations in bone and tissue engineering. Biologia. 2016; 71(4): 353-366.
- 4Dolivo D, Xie P, Sun L, et al. Amnion membranes support wound granulation in a delayed murine excisional wound model. Clin Exp Pharmacol Physiol. 2023; 50(3): 238-246.
- 5Tenenhaus M. The use of dehydrated human amnion/chorion membranes in the treatment of burns and complex wounds: current and future applications. Ann Plast Surg. 2017; 78(2 Suppl 1): S11-s13.
- 6Holtzclaw DJ, Toscano NJJCAiP. Amnion–chorion allograft barrier used for guided tissue regeneration treatment of periodontal intrabony defects: a retrospective observational report. Clinical Advances in Periodontics. 2013; 3(3): 131-137.
10.1902/cap.2012.110110 Google Scholar
- 7Díaz-Prado S, Rendal-Vázquez ME, Muiños-López E, et al. Potential use of the human amniotic membrane as a scaffold in human articular cartilage repair. Cell Tissue Bank. 2010; 11(2): 183-195.
- 8Sarı E, Yalçınozan M, Polat B, Özkayalar H. The effects of cryopreserved human amniotic membrane on fracture healing: animal study. Acta Orthop Traumatol Turc. 2019; 53(6): 485-489.
- 9Jahanafrooz Z, Bakhshandeh B, Behnam Abdollahi S, Seyedjafari E. Human amniotic membrane as a multifunctional biomaterial: recent advances and applications. J Biomater Appl. 2023; 37(8): 1341-1354.
- 10Terada S, Matsuura K, Enosawa S, et al. Inducing proliferation of human amniotic epithelial (HAE) cells for cell therapy. Cell Transplant. 2000; 9(5): 701-704.
- 11Doudi S, Barzegar M, Taghavi EA, et al. Applications of acellular human amniotic membrane in regenerative medicine. Life Sci. 2022; 310:121032.
- 12Rana D, Zreiqat H, Benkirane-Jessel N, Ramakrishna S, Ramalingam M. Development of decellularized scaffolds for stem cell-driven tissue engineering. J Tissue Eng Regen Med. 2017; 11(4): 942-965.
- 13Scarritt ME, Pashos NC, Bunnell BA. A review of cellularization strategies for tissue engineering of whole organs. Front Bioeng Biotechnol. 2015; 3: 43.
- 14Fu X, Liu G, Halim A, Ju Y, Luo Q, Song AG. Mesenchymal stem cell migration and tissue repair. Cell. 2019; 8(8): 784.
- 15Song N, Scholtemeijer M, Shah K. Mesenchymal stem cell immunomodulation: mechanisms and therapeutic potential. Trends Pharmacol Sci. 2020; 41(9): 653-664.
- 16Zhang X, Chen X, Hong H, Hu R, Liu J, Liu C. Decellularized extracellular matrix scaffolds: recent trends and emerging strategies in tissue engineering. Bioact Mater. 2022; 10: 15-31.
- 17Nazari Hashemi P, Chaventre F, Bisson A, Gueudry J, Boyer O, Muraine M. Mapping of proteomic profile and effect of the spongy layer in the human amniotic membrane. Cell Tissue Bank. 2020; 21(2): 329-338.
- 18Avilla-Royo E, Gegenschatz-Schmid K, Grossmann J, et al. Comprehensive quantitative characterization of the human term amnion proteome. Matrix Biology Plus. 2021; 12:100084.
- 19McQuilling JP, Kammer MR, Kimmerling KA, Mowry KC. Characterisation of dehydrated amnion chorion membranes and evaluation of fibroblast and keratinocyte responses in vitro. Int Wound J. 2019; 16(3): 827-840.
- 20Rodríguez-Ares MT, López-Valladares MJ, Touriño R, et al. Effects of lyophilization on human amniotic membrane. Acta Ophthalmol. 2009; 87(4): 396-403.
- 21Russo A, Bonci P, Bonci P. The effects of different preservation processes on the total protein and growth factor content in a new biological product developed from human amniotic membrane. Cell Tissue Bank. 2012; 13(2): 353-361.
- 22Fenelon M, B Maurel D, Siadous R, et al. Comparison of the impact of preservation methods on amniotic membrane properties for tissue engineering applications. Mater Sci Eng C Mater Biol Appl. 2019; 104:109903.
- 23Naseer N, Bashir S, Latief N, Latif F, Khan SN, Riazuddin S. Human amniotic membrane as differentiating matrix for in vitro chondrogenesis. Regen Med. 2018; 13(7): 821-832.
- 24Sous Naasani LI, Rodrigues C, Azevedo JG, Damo Souza AF, Buchner S, Wink MR. Comparison of human denuded amniotic membrane and porcine small intestine submucosa as scaffolds for limbal mesenchymal stem cells. Stem Cell Rev. 2018; 14: 744-754.
- 25Lakkireddy C, Vishwakarma SK, Bardia A, et al. Biofabrication of allogenic bone grafts using cellularized amniotic scaffolds for application in efficient bone healing. Tissue Cell. 2021; 73:101631.
- 26Zafar M, Mirza MR, Awan FR, et al. Effect of APOB polymorphism rs562338 (G/a) on serum proteome of coronary artery disease patients: a “proteogenomic” approach. Sci Rep. 2021; 11(1): 22766.
- 27Mirza MR, Rainer M, Güzel Y, Choudhary IM, Bonn GK. A novel strategy for phosphopeptide enrichment using lanthanide phosphate co-precipitation. Anal Bioanal Chem. 2012; 404(3): 853-862.
- 28Ekram S, Khalid S, Bashir I, Salim A, Khan I. Human umbilical cord-derived mesenchymal stem cells and their chondroprogenitor derivatives reduced pain and inflammation signaling and promote regeneration in a rat intervertebral disc degeneration model. Mol Cell Biochem. 2021; 476(8): 3191-3205.
- 29Peck Justice SA, McCracken NA, Victorino JF, Qi GD, Wijeratne AB, Mosley AL. Boosting detection of low-abundance proteins in thermal proteome profiling experiments by addition of an isobaric trigger channel to TMT multiplexes. Anal Chem. 2021; 93(18): 7000-7010.
- 30Echan LA, Tang HY, Ali-Khan N, Lee KB, Speicher DW. Depletion of multiple high-abundance proteins improves protein profiling capacities of human serum and plasma. Proteomics. 2005; 5(13): 3292-3303.
- 31Sun W, Wu S, Wang X, Zheng D, Gao Y. An analysis of protein abundance suppression in data dependent liquid chromatography and tandem mass spectrometry with tryptic peptide mixtures of five known proteins. Eur J Mass Spectrom. 2005; 11(6): 575-580.
- 32Sandora N, Fitria NA, Kusuma TR, Winarno GA, Tanjunga SF, Wardhana A. Amnion bilayer for dressing and graft replacement for delayed grafting of full-thickness burns; a study in a rat model. PLoS One. 2022; 17(1):e0262007.
- 33Nasiry D, Khalatbary AR, Noori A, Abouhamzeh B, Jamalpoor Z. Accelerated wound healing using three-dimensional amniotic membrane scaffold in combination with adipose-derived stem cells in a diabetic rat model. Tissue Cell. 2023; 82:102098.
- 34Furdova A, Czanner G, Koller J, Vesely P, Furda R, Pridavkova Z. Amniotic membrane application in surgical treatment of conjunctival tumors. Sci Rep. 2023; 13(1): 2835.
- 35Phillips M, Maor E, Rubinsky B. Nonthermal irreversible electroporation for tissue decellularization. J Biomech Eng. 2010; 132(9):091003.
- 36Mohiuddin OA, Campbell B, Poche JN, et al. Decellularized adipose tissue hydrogel promotes bone regeneration in critical-sized mouse femoral defect model. Front Bioeng Biotechnol. 2019; 7: 211.
- 37Li Y, An S, Deng C, Xiao S. Human acellular amniotic membrane as skin substitute and biological scaffold: a review of its preparation, preclinical research, and clinical application. Pharmaceutics. 2023; 15(9): 2249.
- 38Zou Y, Zhang Y. Mechanical evaluation of decellularized porcine thoracic aorta. J Surg Res. 2012; 175(2): 359-368.
- 39Carbonaro D, Putame G, Castaldo C, et al. A low-cost scalable 3D-printed sample-holder for agitation-based decellularization of biological tissues. Med Eng Phys. 2020; 85: 7-15.
- 40Kim JK, Koh YD, Kim JO, Seo DH. Development of a decellularization method to produce nerve allografts using less invasive detergents and hyper/hypotonic solutions. J Plast Reconstr Aesthet Surg. 2016; 69(12): 1690-1696.
- 41Guo Q, Lu X, Xue Y, Zheng H, Zhao X, Zhao H. A new candidate substrate for cell-matrix adhesion study: the acellular human amniotic matrix. J Biomed Biotechnol. 2012; 2012:306083.
- 42Chen C, Zheng S, Zhang X, et al. Transplantation of amniotic scaffold-seeded mesenchymal stem cells and/or endothelial progenitor cells from bone marrow to efficiently repair 3-cm circumferential urethral defect in model dogs. Tissue Eng Part A. 2018; 24(1–2): 47-56.
- 43Shi P, Gao M, Shen Q, Hou L, Zhu Y, Wang J. Biocompatible surgical meshes based on decellularized human amniotic membrane. Mater Sci Eng C Mater Biol Appl. 2015; 54: 112-119.
- 44Amensag S, McFetridge PS. Tuning scaffold mechanics by laminating native extracellular matrix membranes and effects on early cellular remodeling. J Biomed Mater Res A. 2014; 102(5): 1325-1333.
- 45Fenelon M, Etchebarne M, Siadous R, et al. Assessment of fresh and preserved amniotic membrane for guided bone regeneration in mice. J Biomed Mater Res A. 2020; 108(10): 2044-2056.
- 46Kwansa AL, De Vita R, Freeman JW. Tensile mechanical properties of collagen type I and its enzymatic crosslinks. Biophys Chem. 2016; 214-215: 1-10.
- 47Boudko SP, Danylevych N, Hudson BG, Pedchenko VK. Basement membrane collagen IV: isolation of functional domains. Methods in Cell Biology. Elsevier; 2018: 171-185.
- 48Theocharidis G, Drymoussi Z, Kao AP, et al. Type VI collagen regulates dermal matrix assembly and fibroblast motility. J Invest Dermatol. 2016; 136(1): 74-83.
- 49Nyström A, Velati D, Mittapalli VR, Fritsch A, Kern JS, Bruckner-Tuderman L. Collagen VII plays a dual role in wound healing. J Clin Invest. 2013; 123(8): 3498-3509.
- 50Izu Y, Birk DEJFiC, Biology D. Collagen XII mediated cellular and extracellular mechanisms in development, regeneration, and disease. Front. Cell Dev. Biol. 2023; 11:1129000.
- 51Nunes QM, Li Y, Sun C, Kinnunen TK, Fernig DG. Fibroblast growth factors as tissue repair and regeneration therapeutics. PeerJ. 2016; 4:e1535.
- 52Maddaluno L, Urwyler C, Werner S. Fibroblast growth factors: key players in regeneration and tissue repair. Development. 2017; 144(22): 4047-4060.
- 53Bao P, Kodra A, Tomic-Canic M, Golinko MS, Ehrlich HP, Brem H. The role of vascular endothelial growth factor in wound healing. J Surg Res. 2009; 153(2): 347-358.
- 54Kearney KJ, Ariëns RA, Macrae FL. The role of fibrin (ogen) in wound healing and infection control. Seminars in Thrombosis and Hemostasis. Thieme Medical Publishers, Inc; 2021.
- 55Li S, Sampson C, Liu C, Piao HL, Liu HX. Integrin signaling in cancer: bidirectional mechanisms and therapeutic opportunities. Cell Commun. Signaling. 2023; 21(1): 266.
- 56Yang Y, Kim SC, Yu T, et al. Functional roles of p38 mitogen-activated protein kinase in macrophage-mediated inflammatory responses. Mediators Inflamm. 2014; 2014:352371.
- 57Milan PB, Amini N, Joghataei MT, et al. Decellularized human amniotic membrane: from animal models to clinical trials. Methods. 2020; 171: 11-19.
- 58Kakabadze Z, Mardaleishvili K, Loladze G, et al. Clinical application of decellularized and lyophilized human amnion/chorion membrane grafts for closing post-laryngectomy pharyngocutaneous fistulas. J Surg Oncol. 2016; 113(5): 538-543.
- 59Mohiuddin OA, Motherwell JM, Rogers E, et al. Characterization and proteomic analysis of Decellularized adipose tissue hydrogels derived from lean and overweight/obese human donors. Adv Biosyst. 2020; 4(10):e2000124.
- 60Batchelder CA, Martinez ML, Tarantal AFJPo. Natural scaffolds for renal differentiation of human embryonic stem cells for kidney tissue engineering. PloS One. 2015; 10(12):e0143849.
- 61Naasani LIS, Pretto L, Zanatelli C, et al. Bioscaffold developed with decellularized human amniotic membrane seeded with mesenchymal stromal cells: assessment of efficacy and safety profiles in a second-degree burn preclinical model. Biofabrication. 2022; 15(1):015012.
- 62Gholipourmalekabadi M, Sameni M, Radenkovic D, Mozafari M, Mossahebi-Mohammadi M, Seifalian A. Decellularized human amniotic membrane: how viable is it as a delivery system for human adipose tissue-derived stromal cells? Cell Prolif. 2016; 49(1): 115-121.
- 63Davoodi S, Ebrahimpour-Malekshah R, Ayna Ö, et al. Decellularized human amniotic membrane engraftment in combination with adipose-derived stem cells transplantation, synergistically improved diabetic wound healing. J Cosmet Dermatol. 2022; 21(12): 6939-6950.
- 64Diller RB, Tabor AJ. The role of the extracellular matrix (ECM) in wound healing: a review. Biomimetics. 2022; 7(3): 87.
- 65Magatti M, Vertua E, Cargnoni A, Silini A, Parolini O. The immunomodulatory properties of amniotic cells: the two sides of the coin. Cell Transplant. 2018; 27(1): 31-44.
- 66Li WJ, Tuli R, Huang X, Laquerriere P, Tuan RS. Multilineage differentiation of human mesenchymal stem cells in a three-dimensional nanofibrous scaffold. Biomaterials. 2005; 26(25): 5158-5166.
- 67Shariatzadeh S, Shafiee S, Zafari A, et al. Developing a pro-angiogenic placenta derived amniochorionic scaffold with two exposed basement membranes as substrates for cultivating endothelial cells. Sci Rep. 2021; 11(1): 22508.
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