{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T21:43:04Z","timestamp":1791495784901,"version":"4.3.4"},"reference-count":42,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2025,10,17]],"date-time":"2025-10-17T00:00:00Z","timestamp":1760659200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Tiger Wound Care Medical, LLC.","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The human placenta is a complex organ that supports fetal development and is rich in extracellular matrix proteins and growth factors, making it suitable as a biomaterial in wound care. Placenta-derived amnion-only allografts have traditionally been used in the clinic, but they lack the structural and biochemical complexity of the full three-layer placental membrane, which includes the amnion, intermediate, and chorion layers. Advances in tissue engineering have enabled preservation of multiple layers, giving rise to multilayer placental-based Cellular and Acellular Matrix-like Products (CAMPs) such as Full-Thickness (FT; amnion, intermediate, chorion) and ACA (amnion, intermediate, chorion, amnion). Although these advanced CAMPs are increasingly applied clinically, their molecular composition has not been comprehensively defined. This study presents a global proteomic analysis of FT and ACA, complemented by targeted multiplex analysis of soluble proteins and an in vitro angiogenesis assay. Proteomic profiling identified 8908 structural and bioactive components, with 32.5% of proteins associated with tissue repair and remodeling pathways. Multiplex analysis confirmed accessibility of biologically relevant soluble factors. Endothelial tube formation assays further supported biological relevance, demonstrating that soluble proteins in FT and ACA support angiogenesis. These data provide a molecular characterization of multilayer CAMPs and underscore their potential to deliver durable wound coverage while supporting the local microenvironment.<\/jats:p>","DOI":"10.3390\/ijms262010121","type":"journal-article","created":{"date-parts":[[2025,10,17]],"date-time":"2025-10-17T13:07:22Z","timestamp":1760706442000},"page":"10121","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Native Wound-Repair Proteins Retained in Multilayer Placental CAMPs"],"prefix":"10.3390","volume":"26","author":[{"given":"Pragya","family":"Singh","sequence":"first","affiliation":[{"name":"RegenTX Labs LLC, 3463 Magic Dr Ste 315, San Antonio, TX 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shantanu","family":"Guha","sequence":"additional","affiliation":[{"name":"RegenTX Labs LLC, 3463 Magic Dr Ste 315, San Antonio, TX 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2025-3692","authenticated-orcid":false,"given":"Odalis","family":"Landa","sequence":"additional","affiliation":[{"name":"RegenTX Labs LLC, 3463 Magic Dr Ste 315, San Antonio, TX 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-8929-8426","authenticated-orcid":false,"given":"Andrew Ryan","family":"King","sequence":"additional","affiliation":[{"name":"RegenTX Labs LLC, 3463 Magic Dr Ste 315, San Antonio, TX 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2710-040X","authenticated-orcid":false,"given":"Diego","family":"Valdes Cavazos","sequence":"additional","affiliation":[{"name":"RegenTX Labs LLC, 3463 Magic Dr Ste 315, San Antonio, TX 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanna","family":"Marquez","sequence":"additional","affiliation":[{"name":"RegenTX Labs LLC, 3463 Magic Dr Ste 315, San Antonio, TX 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shauna","family":"Hill","sequence":"additional","affiliation":[{"name":"RegenTX Labs LLC, 3463 Magic Dr Ste 315, San Antonio, TX 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Silini, A.R., Cargnoni, A., Magatti, M., Pianta, S., and Parolini, O. (2015). The Long Path of Human Placenta, and Its Derivatives, in Regenerative Medicine. Front. Bioeng. Biotechnol., 3.","DOI":"10.3389\/fbioe.2015.00162"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Protzman, N.M., Mao, Y., Long, D., Sivalenka, R., Gosiewska, A., Hariri, R.J., and Brigido, S.A. (2023). Placental-Derived Biomaterials and Their Application to Wound Healing: A Review. Bioengineering, 10.","DOI":"10.20944\/preprints202306.0211.v1"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"4789","DOI":"10.1039\/D2BM01977A","article-title":"Human placenta\/umbilical cord derivatives in regenerative medicine-Prospects and challenges","volume":"11","author":"Biswas","year":"2023","journal-title":"Biomater. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1700345","DOI":"10.1002\/adhm.201700345","article-title":"Human Perinatal-Derived Biomaterials","volume":"6","author":"Moore","year":"2017","journal-title":"Adv. Healthc. Mater."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"138S","DOI":"10.1097\/PRS.0000000000002643","article-title":"A Review of Cellular and Acellular Matrix Products: Indications, Techniques, and Outcomes","volume":"138","author":"Hughes","year":"2016","journal-title":"Plast. Reconstr. Surg."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"S1","DOI":"10.12968\/jowc.2023.32.Sup4b.S1","article-title":"Best practice for wound repair and regeneration use of cellular, acellular and matrix-like products (CAMPs)","volume":"32","author":"Wu","year":"2023","journal-title":"J. Wound Care"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Roy, A., Mantay, M., Brannan, C., and Griffiths, S. (2022). Placental Tissues as Biomaterials in Regenerative Medicine. Biomed. Res. Int., 2022.","DOI":"10.1155\/2022\/6751456"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Oyen, M.L., Ferguson, V.L., and Calvin, S.E. (2007). Fracture Resistance of Human Amnion. Am. Soceity Mech. Eng., 841\u2013842.","DOI":"10.1115\/SBC2007-174552"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1126","DOI":"10.1002\/term.3086","article-title":"Intermediate layer contribution in placental membrane allografts","volume":"14","author":"Roy","year":"2020","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"12","DOI":"10.21767\/2471-3082.100043","article-title":"Amnion and Chorion Membranes for Root Coverage Procedures: An In Vitro Evaluation of its Physical Characteristics","volume":"4","author":"George","year":"2018","journal-title":"Periodontics Prosthodont."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1177\/1933719111424454","article-title":"Extracellular matrix dynamics and fetal membrane rupture","volume":"20","author":"Strauss","year":"2013","journal-title":"Reprod. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1111\/iwj.12140","article-title":"Biological properties of dehydrated human amnion\/chorion composite graft: Implications for chronic wound healing","volume":"10","author":"Koob","year":"2013","journal-title":"Int. Wound J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1089\/wound.2020.1315","article-title":"Biophysical Characterization of a Novel Tri-Layer Placental Allograft Membrane","volume":"11","author":"Bonvallet","year":"2022","journal-title":"Adv. Wound Care"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Singh, P., Easley, A., Menchaca, K.T., Fanniel, V., Gomez, R., Marquez, J., and Hill, S. (2025). Comparative Study of Placental Allografts with Distinct Layer Composition. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26073406"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e14888","DOI":"10.1111\/iwj.14888","article-title":"Characterization of a full-thickness decellularized and lyophilized human placental membrane for clinical applications","volume":"21","author":"Wetzell","year":"2024","journal-title":"Int. Wound J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1007\/s00441-012-1424-6","article-title":"Amniotic membrane: From structure and functions to clinical applications","volume":"349","author":"Mamede","year":"2012","journal-title":"Cell Tissue Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1016\/j.biomaterials.2009.09.034","article-title":"Preservation, sterilization and de-epithelialization of human amniotic membrane for use in ocular surface reconstruction","volume":"31","author":"Riau","year":"2010","journal-title":"Biomaterials"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12014-022-09388-4","article-title":"A proteomic profile of the healthy human placenta","volume":"20","author":"Manna","year":"2023","journal-title":"Clin. Proteomics"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s13287-021-02684-0","article-title":"Applications of the amniotic membrane in tissue engineering and regeneration: The hundred-year challenge","volume":"13","author":"Elkhenany","year":"2022","journal-title":"Stem Cell Res. Ther."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1002\/jemt.10249","article-title":"Angiogenesis in wound repair: Angiogenic growth factors and the extracellular matrix","volume":"60","author":"Li","year":"2003","journal-title":"Microsc. Res. Tech."},{"key":"ref_21","first-page":"316235","article-title":"Curbing Inflammation in Skin Wound Healing: A Review","volume":"2015","author":"Rosique","year":"2015","journal-title":"Int. J. Inflam."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e70330","DOI":"10.1111\/iwj.70330","article-title":"Reactive Oxygen Species and Antioxidants in Wound Healing: Mechanisms and Therapeutic Potential","volume":"22","author":"Ukaegbu","year":"2025","journal-title":"Int. Wound J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e11","DOI":"10.1016\/j.jid.2016.11.020","article-title":"Research Techniques Made Simple: Analysis of Collective Cell Migration Using the Wound Healing Assay","volume":"137","author":"Grada","year":"2017","journal-title":"J. Investig. Dermatol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1038\/sj.jidsymp.5650001","article-title":"Reactive oxygen species and their detoxification in healing skin wounds","volume":"11","author":"Kumin","year":"2006","journal-title":"J. Investig. Dermatol. Symp. Proc."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1038\/sj.jid.5700701","article-title":"Inflammation in wound repair: Molecular and cellular mechanisms","volume":"127","author":"Eming","year":"2007","journal-title":"J. Investig. Dermatol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1089\/wound.2014.0608","article-title":"The Innate Immune System in Acute and Chronic Wounds","volume":"5","author":"MacLeod","year":"2016","journal-title":"Adv. Wound Care"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3861","DOI":"10.1007\/s00018-016-2268-0","article-title":"Transition from inflammation to proliferation: A critical step during wound healing","volume":"73","author":"Landen","year":"2016","journal-title":"Cell Mol. Life Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1089\/wound.2012.0406","article-title":"Critical Role of Transforming Growth Factor Beta in Different Phases of Wound Healing","volume":"2","author":"Pakyari","year":"2013","journal-title":"Adv. Wound Care"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"344","DOI":"10.1016\/j.addr.2018.06.019","article-title":"Re-epithelialization of adult skin wounds: Cellular mechanisms and therapeutic strategies","volume":"146","author":"Rousselle","year":"2019","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_30","unstructured":"Alhajj, M., and Goyal, A. (2025). Physiology, Granulation Tissue. StatPearls, StatPearls Publishing."},{"key":"ref_31","first-page":"E36","article-title":"Proteomic Comparison of Amnion and Chorion and Evaluation of the Effects of Processing on Placental Membranes","volume":"29","author":"McQuilling","year":"2017","journal-title":"Wounds"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1007\/s10561-020-09821-8","article-title":"Mapping of proteomic profile and effect of the spongy layer in the human amniotic membrane","volume":"21","author":"Chaventre","year":"2020","journal-title":"Cell Tissue Bank."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1133","DOI":"10.1002\/jbm.b.33265","article-title":"Cytokines in single layer amnion allografts compared to multilayer amnion\/chorion allografts for wound healing","volume":"103","author":"Koob","year":"2015","journal-title":"J. Biomed. Mater. Res. B Appl. Biomater."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1111\/wrr.12757","article-title":"A mechanistic evaluation of the angiogenic properties of a dehydrated amnion chorion membrane in vitro and in vivo","volume":"27","author":"McQuilling","year":"2019","journal-title":"Wound Repair. Regen."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1186\/2045-824X-6-10","article-title":"Angiogenic properties of dehydrated human amnion\/chorion allografts: Therapeutic potential for soft tissue repair and regeneration","volume":"6","author":"Koob","year":"2014","journal-title":"Vasc. Cell"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1111\/iwj.12748","article-title":"In vitro assessment of a novel, hypothermically stored amniotic membrane for use in a chronic wound environment","volume":"14","author":"McQuilling","year":"2017","journal-title":"Int. Wound J."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1186\/s13287-021-02192-1","article-title":"Conditioned medium from primary cytotrophoblasts, primary placenta-derived mesenchymal stem cells, or sub-cultured placental tissue promoted HUVEC angiogenesis in vitro","volume":"12","author":"Ma","year":"2021","journal-title":"Stem Cell Res. Ther."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Na, D., Son, H., and Gsponer, J. (2014). Categorizer: A tool to categorize genes into user-defined biological groups based on semantic similarity. BMC Genom., 15.","DOI":"10.1186\/1471-2164-15-1091"},{"key":"ref_39","unstructured":"Institute, E.B. (2025, September 04). QuickGO: Gene Ontology Browser. Available online: https:\/\/www.ebi.ac.uk\/QuickGO\/."},{"key":"ref_40","unstructured":"Consortium, G.O. (2025, September 04). Downloads-Gene Ontology Consortium. Available online: https:\/\/current.geneontology.org\/products\/pages\/downloads.html."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Carpentier, G., Berndt, S., Ferratge, S., Rasband, W., Cuendet, M., Uzan, G., and Albanese, P. (2020). Angiogenesis Analyzer for ImageJ-A comparative morphometric analysis of \u201cEndothelial Tube Formation Assay\u201d and \u201cFibrin Bead Assay\u201d. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-67289-8"},{"key":"ref_42","unstructured":"American Association of Tissue Banks (2016). Standards for Tissue Banking, American Association of Tissue Banks. [14th ed.]."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/20\/10121\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T04:10:58Z","timestamp":1760933458000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/20\/10121"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,17]]},"references-count":42,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["ijms262010121"],"URL":"https:\/\/doi.org\/10.3390\/ijms262010121","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,17]]}}}