{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,22]],"date-time":"2026-01-22T16:41:45Z","timestamp":1769100105176,"version":"3.49.0"},"reference-count":57,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2018,9,19]],"date-time":"2018-09-19T00:00:00Z","timestamp":1537315200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Greige cotton (unbleached cotton) is an intact plant fiber that retains much of the outer cotton fiber layers. These layers contain pectin, peroxidases, and trace metals that are associated with hydrogen peroxide (H2O2) generation during cotton fiber development. When greige cotton is subjected to a nonwoven hydroentanglement process, components of the outer cotton fiber layers are retained. When hydrated, this fabric can generate H2O2 (5\u201350 micromolar). This range has been characterized as inducing accelerated wound healing associated with enhanced cell signaling and the proliferation of cells vital to wound restoration. On the other hand, H2O2 levels above 50 micromolar have been associated with bacteriostatic activity. Here, we report the preparation and hydrogen peroxide activity of copper\/ascorbate formulations, both as adsorbed and in situ synthesized analogs on cotton. The cooper\/ascorbate-cotton formulations were designed with the goal of modulating hydrogen peroxide levels within functional ranges beneficial to wound healing. The cotton\/copper formulation analogs were prepared on nonwoven unbleached cotton and characterized with cotton impregnation titers of 3\u201314 mg copper per gram of cotton. The copper\/ascorbate cotton analog formulations were characterized spectroscopically, and the copper titer was quantified with ICP analysis and probed for peroxide production through assessment with Amplex Red. All analogs demonstrated antibacterial activity. Notably, the treatment of unbleached cotton with low levels of ascorbate (~2 mg\/g cotton) resulted in a 99 percent reduction in Klebsiella pneumoniae and Staphylococcus aureus. In situ synthesized copper\/ascorbate nanoparticles retained activity and did not leach out upon prolonged suspension in an aqueous environment. An assessment of H2O2 effects on fibroblast proliferation are discussed in light of the copper\/cotton analogs and wound healing.<\/jats:p>","DOI":"10.3390\/molecules23092399","type":"journal-article","created":{"date-parts":[[2018,9,19]],"date-time":"2018-09-19T10:50:31Z","timestamp":1537354231000},"page":"2399","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Hydrogen Peroxide Generation of Copper\/Ascorbate Formulations on Cotton: Effect on Antibacterial and Fibroblast Activity for Wound Healing Application"],"prefix":"10.3390","volume":"23","author":[{"given":"J. Vincent","family":"Edwards","sequence":"first","affiliation":[{"name":"Southern Regional Research Center, USDA-ARS, New Orleans, LA 70120, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicolette T.","family":"Prevost","sequence":"additional","affiliation":[{"name":"Southern Regional Research Center, USDA-ARS, New Orleans, LA 70120, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Santiago","sequence":"additional","affiliation":[{"name":"Southern Regional Research Center, USDA-ARS, New Orleans, LA 70120, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Terri","family":"Von Hoven","sequence":"additional","affiliation":[{"name":"Southern Regional Research Center, USDA-ARS, New Orleans, LA 70120, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brian D.","family":"Condon","sequence":"additional","affiliation":[{"name":"Southern Regional Research Center, USDA-ARS, New Orleans, LA 70120, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huzaifah","family":"Qureshi","sequence":"additional","affiliation":[{"name":"Plastic and Reconstructive Surgery, Virginia Commonwealth University, Richmond, VA 23111, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dorne R.","family":"Yager","sequence":"additional","affiliation":[{"name":"Plastic and Reconstructive Surgery, Virginia Commonwealth University, Richmond, VA 23111, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,9,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1533\/9781845696306.1.48","article-title":"Textile materials and structures for wound care products","volume":"Volume 85","author":"Gupta","year":"2009","journal-title":"Advanced Textiles for Wound Care"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1038\/emm.1999.9","article-title":"Redox signaling: Hydrogen peroxide as intracellular messenger","volume":"31","author":"Rhee","year":"1999","journal-title":"Exp. Mol. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.1016\/j.bbagen.2008.01.006","article-title":"Redox signals in wound healing","volume":"1780","author":"Sen","year":"2008","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.ymthe.2005.07.684","article-title":"Dermal wound healing is subject to redox control","volume":"13","author":"Roy","year":"2006","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1038\/nature08119","article-title":"A tissue-scale gradient of hydrogen peroxide mediates rapid wound detection in zebrafish","volume":"459","author":"Niethammer","year":"2009","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1159\/000475501","article-title":"Hydrogen peroxide: A potential wound therapeutic target?","volume":"26","author":"Zhu","year":"2017","journal-title":"Med. Princ. Pract."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1083\/jcb.201102095","article-title":"Signal transduction by reactive oxygen species","volume":"194","author":"Finkel","year":"2011","journal-title":"J. Cell Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1111\/j.1365-2133.1993.tb03518.x","article-title":"Quantification of hydrogen peroxide generation by granuflex (DuoDERM) hydrocolloid granules and its constituents (gelatin, sodium carboxymethylcellulose, and pectin)","volume":"129","author":"Schmidt","year":"1993","journal-title":"Br. J. Dermatol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"14908","DOI":"10.1038\/srep14908","article-title":"Electrochemical scaffold generates localized, low concentration of hydrogen peroxide that inhibits bacterial pathogens and biofilms","volume":"5","author":"Sultana","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Banerjee, J., Ghatak, P.D., Roy, S., Khanna, S., Hemann, C., Deng, B., Das, A., Zweier, J.L., Wozniak, D., and Sen, C.K. (2015). Silver-zinc redox-coupled electroceutical wound dressing disrupts bacterial biofilm. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0119531"},{"key":"ref_11","first-page":"50","article-title":"Efficacy of a bio-electric dressing in healing deep, partial-thickness wounds using a porcine model","volume":"58","author":"Harding","year":"2012","journal-title":"Ostomy Wound Manag."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3","DOI":"10.7150\/jbji.16690","article-title":"Hydrogen peroxide wound irrigation in orthopaedic surgery","volume":"2","author":"Lu","year":"2017","journal-title":"J. Bone Jt. Infect."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1089\/10755530360623383","article-title":"The effect of dilution on the rate of hydrogen peroxide production in honey and its implications for wound healing","volume":"9","author":"Bang","year":"2003","journal-title":"J. Altern. Complement. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1016\/S1043-4666(03)00092-9","article-title":"Honey stimulates inflammatory cytokine production from monocytes","volume":"21","author":"Tonks","year":"2003","journal-title":"Cytokine"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Edwards, J.V., Prevost, N.T., Nam, S., Hinchliffe, D., Condon, B., and Yager, D. (2017). Induction of low-level hydrogen peroxide generation by unbleached cotton nonwovens as potential wound dressing materials. J. Funct. Biomater., 8.","DOI":"10.3390\/jfb8010009"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1042\/bj3320507","article-title":"Oxidative scission of plant cell wall polysaccharides by ascorbate-induced hydroxyl radicals","volume":"332","author":"Fry","year":"1998","journal-title":"Biochem. J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/S0031-9422(99)00586-5","article-title":"Scission of polysaccharides by peroxidase-generated hydroxyl radicals","volume":"53","author":"Schweikert","year":"2000","journal-title":"Phytochemistry"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1007\/s00425-008-0734-0","article-title":"Cu\/Zn superoxide dismutases in developing cotton fibers: Evidence for an extracellular form","volume":"228","author":"Kim","year":"2008","journal-title":"Planta"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"195","DOI":"10.3349\/ymj.2017.58.1.195","article-title":"Efficacy of oxidized regenerated cellulose, surgiguard\u00ae, in porcine surgery","volume":"58","author":"Kim","year":"2017","journal-title":"Yonsei Med. J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1021\/bm010008g","article-title":"Molecular basis of C(2+)-induced gelation in alginates and pectins: The egg-box model revisited","volume":"2","author":"Braccini","year":"2001","journal-title":"Biomacromolecules"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1177\/1528083708092012","article-title":"Development of a continuous finishing chemistry process for manufacture of a phosphorylated cotton chronic wound dressing","volume":"39","author":"Edwards","year":"2009","journal-title":"J. Ind. Text."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"900","DOI":"10.1039\/c0gc00772b","article-title":"Synthesis of highly stable dispersions of nanosized copper particles using l-ascorbic acid","volume":"13","author":"Xiong","year":"2011","journal-title":"Green Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.watres.2014.06.022","article-title":"Incorporation of copper nanoparticles into paper for point-of-use water purification","volume":"63","author":"Dankovich","year":"2014","journal-title":"Water Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1007\/s11051-005-9065-2","article-title":"One-step green route to narrowly dispersed copper nanocrystals","volume":"8","author":"Wu","year":"2006","journal-title":"J. Nanopart. Res."},{"key":"ref_25","first-page":"143","article-title":"Dependence of spectroscopic properties of copper oxide based silica supported nanostructure on temperature","volume":"8","author":"Tohidi","year":"2012","journal-title":"Nano Sci. Technol. Int."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"532","DOI":"10.2307\/3579271","article-title":"Catalytic metals, ascorbate and free radicals: Combinations to avoid","volume":"145","author":"Buettner","year":"1996","journal-title":"Radiat. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/0165-022X(88)90100-5","article-title":"In the absence of catalytic metals ascorbate does not autoxidize at pH 7: Ascorbate as a test for catalytic metals","volume":"16","author":"Buettner","year":"1988","journal-title":"J. Biochem. Biophys. Methods"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"38541","DOI":"10.1039\/C6RA02843H","article-title":"Generation of hydrogen peroxide and hydroxyl radical resulting from oxygen-dependent oxidation of l-ascorbic acid via copper redox-catalyzed reactions","volume":"6","author":"Zhou","year":"2016","journal-title":"RSC Adv."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2963","DOI":"10.1007\/s10570-014-0270-y","article-title":"Internally dispersed synthesis of uniform silver nanoparticles via in situ reduction of [Ag(NH3)2]+ along natural microfibrillar substructures of cotton fiber","volume":"21","author":"Nam","year":"2014","journal-title":"Cellulose"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1177\/0040517511416276","article-title":"Effect of water pressure on absorbency of hydroentangled greige cotton non-woven fabrics","volume":"82","author":"Sawhney","year":"2012","journal-title":"Text. Res. J."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1111\/iwj.12557","article-title":"Reactive oxygen species (ROS) and wound healing: The functional role of ROS and emerging ROS-modulating technologies for augmentation of the healing process","volume":"14","author":"Dunnill","year":"2017","journal-title":"Int. Wound J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1723","DOI":"10.1167\/iovs.10-5866","article-title":"Low levels of hydrogen peroxide stimulate corneal epithelial cell adhesion, migration, and wound healing","volume":"52","author":"Pan","year":"2011","journal-title":"Investig. Ophthalmol. Visual Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1233","DOI":"10.4315\/0362-028X-59.11.1233","article-title":"Antibacterial effects of hydrogen peroxide and methods for its detection and quantitation","volume":"59","author":"Juven","year":"1996","journal-title":"J. Food Prot."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"721","DOI":"10.7150\/thno.9052","article-title":"Luminescent dual sensors reveal extracellular pH-gradients and hypoxia on chronic wounds that disrupt epidermal repair","volume":"4","author":"Schreml","year":"2014","journal-title":"Theranostics"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2119","DOI":"10.1007\/s10570-014-0215-5","article-title":"Copper nanoparticles on bleached cotton fabric: In situ synthesis and characterization","volume":"21","author":"Sedighi","year":"2014","journal-title":"Cellulose"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2506","DOI":"10.1002\/adfm.201100123","article-title":"Copper-based nanostructured coatings on natural cellulose: Nanocomposites exhibiting rapid and efficient inhibition of a multi-drug resistant wound pathogen, A. baumannii, and mammalian cell biocompatibility in vitro","volume":"21","author":"Cady","year":"2011","journal-title":"Adv. Funct. Mater."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1186\/s11671-016-1240-0","article-title":"Antibacterial and antimycotic activity of cotton fabrics, impregnated with silver and binary silver\/copper nanoparticles","volume":"11","author":"Eremenko","year":"2016","journal-title":"Nanoscale Res. Lett."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"135101","DOI":"10.1088\/0957-4484\/25\/13\/135101","article-title":"Mechanism of antibacterial activity of copper nanoparticles","volume":"25","author":"Chatterjee","year":"2014","journal-title":"Nanotechnology"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"88","DOI":"10.4103\/jcrsm.jcrsm_35_17","article-title":"Antimicrobial activity of vitamin c demonstrated on uropathogenic Escherichia coli and Klebsiella pneumoniae","volume":"3","author":"Verghese","year":"2017","journal-title":"J. Curr. Res. Sci. Med."},{"key":"ref_40","first-page":"32","article-title":"Formaldehyde-free durable-press finishes","volume":"22","author":"Welch","year":"1992","journal-title":"Rev. Prog. Color. Relat. Top."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1021\/jp209542x","article-title":"Crystallographic verification that copper(II) coordinates to four of the oxygen atoms of zeolite 6-rings. Two single-crystal structures of fully dehydrated, largely Cu2+-exchanged zeolite Y (FAU, Si\/Al = 1.56)","volume":"116","author":"Seo","year":"2012","journal-title":"J. Phys. Chem. C"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1016\/j.jphotochem.2005.12.020","article-title":"Performance and characterization of Ag\u2013cotton and Ag\/TiO2 loaded textiles during the abatement of E. coli","volume":"181","author":"Yuranova","year":"2006","journal-title":"J. Photochem. Photobiol. A Chem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2269","DOI":"10.1016\/j.freeradbiomed.2012.10.538","article-title":"Reactive oxygen species-dependent wound responses in animals and plants","volume":"53","author":"Suzuki","year":"2012","journal-title":"Free Radic. Biol. Med."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.bbrc.2012.05.100","article-title":"Effects of hydrogen peroxide in a keratinocyte-fibroblast co-culture model of wound healing","volume":"423","author":"Loo","year":"2012","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.pharmthera.2009.02.005","article-title":"Regulation of cell proliferation by nadph oxidase-mediated signaling: Potential roles in tissue repair, regenerative medicine and tissue engineering","volume":"122","author":"Chan","year":"2009","journal-title":"Pharmacol. Ther."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1159\/000455548","article-title":"Effect of copper on menkes\u2019 and normal cultured skin fibroblasts","volume":"1","author":"Beratis","year":"1980","journal-title":"Dev. Pharmacol. Ther."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1007\/s10544-012-9651-x","article-title":"Concentration-dependent cytotoxicity of copper ions on mouse fibroblasts in vitro: Effects of copper ion release from TCu380A vs. TCu220C intra-uterine devices","volume":"14","author":"Cao","year":"2012","journal-title":"Biomed. Microdevices"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1111\/j.1751-1097.1979.tb07128.x","article-title":"Action of hydrogen peroxide on human fibroblast in culture","volume":"30","author":"Hoffmann","year":"1979","journal-title":"Photochem. Photobiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1016\/j.jdermsci.2016.09.001","article-title":"Human fibroblasts treated with hydrogen peroxide stimulate human melanoblast proliferation and melanocyte differentiation, but inhibit melanocyte proliferation in serum-free co-culture system","volume":"84","author":"Hirobe","year":"2016","journal-title":"J. Dermatol. Sci."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2977","DOI":"10.1016\/j.bbamcr.2016.09.012","article-title":"Activation of apoptosis signalling pathways by reactive oxygen species","volume":"1863","year":"2016","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Rieger, S., and Sagasti, A. (2011). Hydrogen peroxide promotes injury-induced peripheral sensory axon regeneration in the zebrafish skin. PLoS Biol., 9.","DOI":"10.1371\/journal.pbio.1000621"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1615\/PlasmaMed.2016015738","article-title":"In vitro and in vivo analysis of hydrogen peroxide-enhanced plasma-induced effluent for infection and contamination mitigation at research and medical facilities","volume":"5","author":"Golkowski","year":"2015","journal-title":"Plasma Med."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1021\/bk-2001-0792.ch012","article-title":"Determining antimicrobial efficacy and biocompatibility of treated articles using standard test methods","volume":"Volume 792","author":"Joiner","year":"2001","journal-title":"Bioactive Fibers and Polymers"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"379","DOI":"10.4103\/0970-0358.101321","article-title":"Recent advances in topical wound care","volume":"45","author":"Sarabahi","year":"2012","journal-title":"Indian J. Plast. Surg."},{"key":"ref_55","unstructured":"Elliott, I.M.Z., and Elliott, J.R. (1964). A Short History of Surgical Dressings, Pharmaceutical Press."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1089\/wound.2014.0586","article-title":"Choosing a wound dressing based on common wound characteristics","volume":"5","author":"Dabiri","year":"2016","journal-title":"Adv. Wound Care"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1007\/s12325-017-0478-y","article-title":"Chronic wound healing: A review of current management and treatments","volume":"34","author":"Han","year":"2017","journal-title":"Adv. Ther."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/23\/9\/2399\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:21:27Z","timestamp":1760196087000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/23\/9\/2399"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,9,19]]},"references-count":57,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2018,9]]}},"alternative-id":["molecules23092399"],"URL":"https:\/\/doi.org\/10.3390\/molecules23092399","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,9,19]]}}}