{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T07:18:09Z","timestamp":1787987889545,"version":"build-2784847793"},"reference-count":43,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,2,1]],"date-time":"2025-02-01T00:00:00Z","timestamp":1738368000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Universidad de La Sabana","award":["MED 291-2020"],"award-info":[{"award-number":["MED 291-2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["MPs"],"abstract":"<jats:p>The human amniotic membrane (hAM) has been used as an implant to enhance the regenerative process and control inflammation in different diseases, given their structure, biocompatibility, presence of stem cells and multiple growth factors. The objective of this study was to generate a standardized protocol for obtaining, processing, and storing hAMs that guarantee the conservation of their structural and cellular characteristics as well as their mechanical properties, ensuring their ease of handling, sterility, and quality that allows their implementation for therapeutic purposes in the field of regenerative medicine. The hAMs were obtained from mothers with healthy, full-term, controlled pregnancies and by cesarean section. The hAMs were processed under sterile conditions, manually separated from the placenta and, subsequently, they were frozen in a solution of culture medium plus 50% v\/v glycerol. The protocol allows obtaining sterile hAMs composed of both epithelium and stroma with adequate preservation of the amniotic cells. The glycerol\u2019s impact on the mechanical properties may enhance the membrane\u2019s adaptability and conformability to diverse wound surfaces, potentially improving the healing process. It is necessary to repeat microbiological, cell viability and mechanical studies at 6 and 12 months to ensure that long-term frozen conditions do not affect the quality of the hAMs.<\/jats:p>","DOI":"10.3390\/mps8010012","type":"journal-article","created":{"date-parts":[[2025,2,3]],"date-time":"2025-02-03T04:36:51Z","timestamp":1738557411000},"page":"12","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Procurement, Processing, and Storage of Human Amniotic Membranes for Implantation Purposes in Non-Healing Pressure Ulcers"],"prefix":"10.3390","volume":"8","author":[{"given":"Lina A.","family":"G\u00f3mez","sequence":"first","affiliation":[{"name":"Biomedical Research Center (CIBUS), Faculty of Medicine, Universidad de La Sabana, Ch\u00eda 53753, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2068-8530","authenticated-orcid":false,"given":"Carlos","family":"Dom\u00ednguez-Paz","sequence":"additional","affiliation":[{"name":"Department of Prototypes and Manufacturing, Grupo de Energia, Materiales y Ambiente (GEMA), Universidad de La Sabana, Ch\u00eda 140013, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan F.","family":"Ospina","sequence":"additional","affiliation":[{"name":"Hospital Universitario La Samaritana, Ginecohus Research Group, Bogot\u00e1 111321, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5254-2256","authenticated-orcid":false,"given":"Elga J.","family":"Vargas","sequence":"additional","affiliation":[{"name":"Pathology Laboratory, Department of Pathology, Faculty of Medicine, Universidad de La Sabana, Ch\u00eda 140013, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1007\/BF00317947","article-title":"Intercellular junctions in the human fetal membranes","volume":"166","author":"Bartels","year":"1983","journal-title":"Anat. Embryol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/0002-9378(64)90264-9","article-title":"Fetal weight and membrane tensile strength","volume":"88","author":"Polishuk","year":"1964","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1095\/biolreprod.106.055244","article-title":"Stem cells derived from human fetal membranes display multilineage differentiation potential","volume":"77","author":"Ilancheran","year":"2007","journal-title":"Biol. Reprod."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0143-4004(05)80244-9","article-title":"Immunohistochemical localization of insulin-like growth factors (IGFs) and IGF binding proteins-1, -2 and -3 in human placenta and fetal membranes","volume":"14","author":"Hill","year":"1993","journal-title":"Placenta"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Pandansari, P., Tantin, R.D., Abbas, B., and Hilmy, N. (2017). Biological Properties and Functions of Amnion. Human Amniotic Membrane: Basic Science and Clinical Application, World Scientific Publishing Company.","DOI":"10.1142\/9789813226357_0006"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"111","DOI":"10.5500\/wjt.v4.i2.111","article-title":"Human amniotic membrane transplantation: Different modalities of its use in ophthalmology","volume":"4","author":"Malhotra","year":"2014","journal-title":"World J. Transplant."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.placenta.2006.01.006","article-title":"Expression of Natural Antimicrobials by Human Placenta and Fetal Membranes","volume":"28","author":"King","year":"2007","journal-title":"Placenta"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1016\/j.survophthal.2017.09.005","article-title":"Treatment of acute ocular chemical burns","volume":"63","author":"Sharma","year":"2018","journal-title":"Surv. Ophthalmol."},{"key":"ref_9","first-page":"101","article-title":"Transplante de membrana amni\u00f3tica en patolog\u00eda de la superficie ocular","volume":"12","year":"2001","journal-title":"Arch. Soc. Canar. Oftalmol."},{"key":"ref_10","first-page":"82","article-title":"Membrana amni\u00f3tica, revisi\u00f3n de su uso oftalmol\u00f3gico y resultados en los \u00faltimos cinco a\u00f1os (2013\u20132017) en Granada. Estudio preliminar","volume":"103","year":"2018","journal-title":"Actual. Medica"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.cpm.2014.09.002","article-title":"The Use of Human Amnion\/Chorion Membrane in the Clinical Setting for Lower Extremity Repair: A Review","volume":"32","author":"Zelen","year":"2015","journal-title":"Clin. Podiatr. Med. Surg."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.1186\/s40064-016-3107-3","article-title":"Epidemiological and cost analysis of burn injuries admitted to the emergency department of a tertiary burn center","volume":"5","author":"Eser","year":"2016","journal-title":"Springerplus"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.1177\/1071100718788549","article-title":"Randomized Controlled Trial of Micronized Dehydrated Human Amnion\/Chorion Membrane (dHACM) Injection Compared to Placebo for the Treatment of Plantar Fasciitis","volume":"39","author":"Cazzell","year":"2018","journal-title":"Foot Ankle Int."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1136\/bmj.1.4134.517","article-title":"A New Method of Preventing Adhesions the Use of Amnioplastin After Craniotomy","volume":"1","author":"Penfield","year":"1940","journal-title":"Br. Med. J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"S4","DOI":"10.12968\/jowc.2019.28.Sup5.S4","article-title":"An evaluation of dehydrated human amnion\/chorion membrane allografts for pressure ulcer treatment: A case series","volume":"28","author":"Berhane","year":"2019","journal-title":"J. Wound Care"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"249","DOI":"10.18869\/acadpub.beat.5.4.452","article-title":"Grafting with Cryopreserved Amniotic Membrane versus Conservative Wound Care in Treatment of Pressure Ulcers: A Randomized Clinical Trial","volume":"5","author":"Ankvmhee","year":"2017","journal-title":"Bull. Emerg. Trauma."},{"key":"ref_17","first-page":"475","article-title":"Membranas amni\u00f3ticas y su aplicaci\u00f3n en el tratamiento de quemaduras: Una revisi\u00f3n sistem\u00e1tica exploratoria","volume":"75","author":"Hpjspkgskgrl","year":"2023","journal-title":"Rev. Cir."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1137","DOI":"10.1016\/j.burns.2013.01.017","article-title":"Effect of amniotic membrane on graft take in extremity burns","volume":"39","author":"Mohammadi","year":"2013","journal-title":"Burns"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2551","DOI":"10.1097\/SCS.0000000000005826","article-title":"The Use of Dehydrated Human Amniotic\/Chorionic Membrane Skin Substitute in the Treatment of Pediatric Facial Burn","volume":"30","author":"Puyana","year":"2019","journal-title":"J. Craniofacial Surg."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1097\/01.ASW.0000695752.52235.e3","article-title":"Evaluation of Dried Amniotic Membrane on Wound Healing at Split-Thickness Skin Graft Donor Sites: A Randomized, Placebo-Controlled, Double-blind Trial","volume":"33","author":"Vaheb","year":"2020","journal-title":"Adv. Skin. Wound Care"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1774","DOI":"10.1007\/s43032-021-00525-2","article-title":"Application of Tissue Engineering and Regenerative Medicine in Prelabor Rupture of Membranes: A Review of the Current Evidence","volume":"28","author":"Jung","year":"2021","journal-title":"Reprod. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1089\/gyn.1992.8.25","article-title":"Vaginoplasty using Amniotic Membranes in Cases of Vaginal Agenesis or after Vaginectomy","volume":"8","author":"Nisolle","year":"1992","journal-title":"J. Gynecol. Surg."},{"key":"ref_23","unstructured":"Ministerio De Protecci\u00f3n Social (2005). Manual de Buenas Pr\u00e1cticas para Bancos de Tejidos y de M\u00e9dula \u00d3sea y se dictan otras disposiciones. RESOLUCI\u00d3N 5108 COLOMBIA: MINISTERIO DE PROTECCI\u00d3N SOCIAL, Presidencia de la Rep\u00fablica de Colombia."},{"key":"ref_24","first-page":"290","article-title":"Human amniotic membrane as a versatile biological dressing--a preliminary report","volume":"37","author":"Kasi","year":"1987","journal-title":"J. Pak. Med. Assoc."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1016\/S0002-9394(14)71693-9","article-title":"Amniotic membrane transplantation for conjunctival surface reconstruction","volume":"124","author":"Tseng","year":"1997","journal-title":"Am. J. Ophthalmol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.1007\/s00417-018-3973-1","article-title":"Cryopreservation of amniotic membrane with and without glycerol additive","volume":"256","author":"Wagner","year":"2018","journal-title":"Graefe\u2019s Arch. Clin. Exp. Ophthalmol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Ingraldi, A.L., Audet, R.G., and Tabor, A.J. (2023). The Preparation and Clinical Efficacy of Amnion-Derived Membranes: A Review. VolJ. Funct. Biomater., 14.","DOI":"10.3390\/jfb14100531"},{"key":"ref_28","unstructured":"(2024). Title 21-Food and Drugs Chapter I-Food and Drug Administration, Department of Health and Human Services Subchapter L-Regulations Under Certain Other Acts Administered by the Food and Drug Administration, FDA."},{"key":"ref_29","unstructured":"(2025, January 11). Ministerio de Protecci\u00f3n Social, DECRETO N\u00daMERO 2493 DE 2004; Presidencia de la Rep\u00fablica de Colombia: Bogot\u00e1, Colombia, 4 August 2004, Available online: https:\/\/www.minsalud.gov.co\/Normatividad_Nuevo\/DECRETO%202493%20DE%202004.pdf."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1254","DOI":"10.1016\/j.burns.2019.07.005","article-title":"How preparation and preservation procedures affect the properties of amniotic membrane? How safe are the procedures?","volume":"46","author":"Gholipourmalekabadi","year":"2020","journal-title":"Burns"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1198","DOI":"10.1002\/jbm.b.34782","article-title":"Human Amniotic Membrane: A review on tissue engineering, application, and storage","volume":"109","author":"Kern","year":"2021","journal-title":"J. Biomed. Mater. Res. Part B Appl. Biomater."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Pogozhykh, O., Hofmann, N., Gryshkov, O., von Kaisenberg, C., Mueller, M., Glasmacher, B., Pogozhykh, D., B\u00f6rgel, M., Blasczyk, R., and Figueiredo, C. (2020). Repeated freezing procedures preserve structural and functional properties of amniotic membrane for application in ophthalmology. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21114029"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1007\/s10561-023-10100-5","article-title":"Cryopreserved amniotic membrane in chronic nonhealing wounds: A series of case reports","volume":"25","author":"Horvath","year":"2023","journal-title":"Cell Tissue Bank."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1016\/S0305-4179(02)00304-2","article-title":"\u201cAmnion Bank\u201d\u2013The use of long-term glycerol preserved amniotic membranes in the management of superficial and superficial partial thickness burns","volume":"29","author":"Ravishanker","year":"2003","journal-title":"Burns"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1016\/j.survophthal.2009.04.011","article-title":"The Use of Vital Dyes in Ocular Surgery","volume":"54","author":"Rodrigues","year":"2009","journal-title":"Surv. Ophthalmol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1007\/s10561-014-9437-x","article-title":"Storage and qualification of viable intact human amniotic graft and technology transfer to a tissue bank","volume":"15","author":"Laurent","year":"2014","journal-title":"Cell Tissue Bank."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1016\/S0305-4179(99)00072-8","article-title":"Effectiveness of human amnion preserved long-term in glycerol as a temporary biological dressing","volume":"25","author":"Maral","year":"1999","journal-title":"Burns"},{"key":"ref_38","first-page":"115","article-title":"Preparaci\u00f3n en glicerina: Una t\u00e9cnica para la conservaci\u00f3n prolongada de cuerpos en anatom\u00eda veterinaria","volume":"26","author":"Ortiz","year":"2013","journal-title":"Rev. Med. Vet."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"C125","DOI":"10.1152\/ajpcell.1984.247.3.C125","article-title":"Freezing of living cells: Mechanisms and implications","volume":"247","author":"Mazur","year":"1984","journal-title":"Am. J. Physiol. Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1177\/08853282221137609","article-title":"Human amniotic membrane as a multifunctional biomaterial: Recent advances and applications","volume":"37","author":"Jahanafrooz","year":"2023","journal-title":"J. Biomater. Appl."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2226","DOI":"10.1021\/acsbiomaterials.8b00015","article-title":"Human Amniotic Membrane: A Versatile Scaffold for Tissue Engineering","volume":"4","author":"Arrizabalaga","year":"2018","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1053\/j.otsm.2016.12.004","article-title":"Amniotic Fluid, Cells, and Membrane Application","volume":"25","author":"Friel","year":"2017","journal-title":"Oper. Tech. Sports Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e2371","DOI":"10.1097\/GOX.0000000000002371","article-title":"Placental Membrane Provides Improved Healing Efficacy and Lower Cost Versus a Tissue-Engineered Human Skin in the Treatment of Diabetic Foot Ulcerations","volume":"7","author":"Glat","year":"2019","journal-title":"Plast. Reconstr. Surg. Glob. Open"}],"container-title":["Methods and Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2409-9279\/8\/1\/12\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:25:32Z","timestamp":1760027132000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2409-9279\/8\/1\/12"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,1]]},"references-count":43,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["mps8010012"],"URL":"https:\/\/doi.org\/10.3390\/mps8010012","relation":{},"ISSN":["2409-9279"],"issn-type":[{"value":"2409-9279","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,1]]}}}