{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T14:58:39Z","timestamp":1790089119881,"version":"4.0.1"},"reference-count":48,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2022,7,30]],"date-time":"2022-07-30T00:00:00Z","timestamp":1659139200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Postgraduate Innovative Project of Central South University"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The cryopreservation of red blood cells (RBCs) plays a key role in blood transfusion therapy. Traditional cryoprotectants (CPAs) are mostly organic solvents and may cause side effects to RBCs, such as hemolysis and membrane damage. Therefore, it is necessary to find CPAs with a better performance and lower toxicity. Herein, we report for the first time that N-[Tri(hydroxymethyl)methyl]glycine (tricine) showed a great potential in the cryopreservation of sheep RBCs. The addition of tricine significantly increased the thawed RBCs\u2019 recovery from 19.5 \u00b1 1.8% to 81.2 \u00b1 8.5%. The properties of thawed RBCs were also maintained normally. Through mathematical modeling analysis, tricine showed a great efficiency in cryopreservation. We found that tricine had a good osmotic regulation capacity, which could mitigate the dehydration of RBCs during cryopreservation. In addition, tricine inhibited ice recrystallization, thereby decreasing the mechanical damage from ice. Tricine could also reduce oxidative damage during freezing and thawing by scavenging reactive oxygen species (ROS) and maintaining the activities of endogenous antioxidant enzymes. This work is expected to open up a new path for the study of novel CPAs and promote the development of cryopreservation of RBCs.<\/jats:p>","DOI":"10.3390\/ijms23158462","type":"journal-article","created":{"date-parts":[[2022,8,1]],"date-time":"2022-08-01T02:06:42Z","timestamp":1659319602000},"page":"8462","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Tricine as a Novel Cryoprotectant with Osmotic Regulation, Ice Recrystallization Inhibition and Antioxidant Properties for Cryopreservation of Red Blood Cells"],"prefix":"10.3390","volume":"23","author":[{"given":"Xiangjian","family":"Liu","sequence":"first","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuying","family":"Hu","sequence":"additional","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenqian","family":"Zhang","sequence":"additional","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deyi","family":"Yang","sequence":"additional","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxin","family":"Pan","sequence":"additional","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3330-0769","authenticated-orcid":false,"given":"Marlene Davis","family":"Ekpo","sequence":"additional","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingxian","family":"Xie","sequence":"additional","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Zhao","sequence":"additional","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0698-4410","authenticated-orcid":false,"given":"Songwen","family":"Tan","sequence":"additional","affiliation":[{"name":"Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1845","DOI":"10.1016\/S0140-6736(13)60650-9","article-title":"Concepts of blood transfusion in adults","volume":"381","author":"Goodnough","year":"2013","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1016\/j.athoracsur.2018.08.073","article-title":"Real Age: Red Blood Cell Aging During Storage","volume":"107","author":"Koch","year":"2019","journal-title":"Ann. Thorac. Surg."},{"key":"ref_3","first-page":"112","article-title":"The controversy over the age of blood: What do the clinical trials really teach us?","volume":"15","author":"Belpulsi","year":"2017","journal-title":"Blood Transfus."},{"key":"ref_4","first-page":"267","article-title":"The relation between aged blood products and onset of transfusion-related acute lung injury. A review of pre-clinical data","volume":"57","author":"Vlaar","year":"2011","journal-title":"Clin. Lab."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1097\/CCM.0b013e3182711b38","article-title":"Resuscitation with Aged Blood Exacerbates Liver Injury in a Hemorrhagic Rat Model *","volume":"41","author":"Matot","year":"2013","journal-title":"Crit. Care Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1184","DOI":"10.1111\/j.1537-2995.2011.03466.x","article-title":"Transfusion of older stored blood and risk of death: A meta-analysis","volume":"52","author":"Wang","year":"2012","journal-title":"Transfusion"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2801","DOI":"10.1002\/anie.201511454","article-title":"Combining Biomimetic Block Copolymer Worms with an Ice-Inhibiting Polymer for the Solvent-Free Cryopreservation of Red Blood Cells","volume":"55","author":"Mitchell","year":"2016","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1021\/acsbiomaterials.8b01286","article-title":"Betaine Combined with Membrane Stabilizers Enables Solvent-Free Whole Blood Cryopreservation and One-Step Cryoprotectant Removal","volume":"5","author":"Sui","year":"2019","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/0014-4827(72)90303-5","article-title":"A two-factor hypothesis of freezing injury: Evidence from Chinese hamster tissue-culture cells","volume":"71","author":"Mazur","year":"1972","journal-title":"Exp. Cell Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"42516","DOI":"10.1021\/acsami.7b12189","article-title":"Exploring the Potential of Biocompatible Osmoprotectants as Highly Efficient Cryoprotectants","volume":"9","author":"Yang","year":"2017","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1021\/acsmacrolett.9b00386","article-title":"Enhancement of Macromolecular Ice Recrystallization Inhibition Activity by Exploiting Depletion Forces","volume":"8","author":"Ishibe","year":"2019","journal-title":"ACS Macro Lett."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"BSR20191601","DOI":"10.1042\/BSR20191601","article-title":"The roles of reactive oxygen species and antioxidants in cryopreservation","volume":"39","author":"Len","year":"2019","journal-title":"Biosci. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"648684","DOI":"10.3389\/fchem.2021.648684","article-title":"The Feasibility of Antioxidants Avoiding Oxidative Damages from Reactive Oxygen Species in Cryopreservation","volume":"9","author":"Liu","year":"2021","journal-title":"Front. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/S0011-2240(02)00124-4","article-title":"Biochemical stabilization enhances red blood cell recovery and stability following cryopreservation","volume":"45","author":"Wagner","year":"2002","journal-title":"Cryobiology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1039\/D0TB02426K","article-title":"Fine-tuned dehydration by trehalose enables the cryopreservation of RBCs with unusually low concentrations of glycerol","volume":"9","author":"Shen","year":"2021","journal-title":"J. Mater. Chem. B"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.biomaterials.2017.06.018","article-title":"Apatite nanoparticles strongly improve red blood cell cryopreservation by mediating trehalose delivery via enhanced membrane permeation","volume":"140","author":"Stefanic","year":"2017","journal-title":"Biomaterials"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.transci.2012.01.007","article-title":"Hydroxyethylstarch in cryopreservation\u2013Mechanisms, benefits and problems","volume":"46","author":"Stolzing","year":"2012","journal-title":"Transfus. Apher. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1016\/0011-2240(71)90001-0","article-title":"The preservation of erythrocytes at liquid nitrogen temperatures with hydroxyethyl starch: The removal of hydroxyethyl starch from erythrocytes after thawing","volume":"8","author":"Knorpp","year":"1971","journal-title":"Cryobiology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"7789","DOI":"10.1021\/jacs.9b00992","article-title":"Metal\u2013Organic Framework Nanoparticle-Assisted Cryopreservation of Red Blood Cells","volume":"141","author":"Zhu","year":"2019","journal-title":"J. Am. Chem. Soc."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1021\/acs.biomac.1c00599","article-title":"Red Blood Cell Cryopreservation with Minimal Post-Thaw Lysis Enabled by a Synergistic Combination of a Cryoprotecting Polyampholyte with DMSO\/Trehalose","volume":"23","author":"Murray","year":"2022","journal-title":"Biomacromolecules"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"100156","DOI":"10.1016\/j.mtbio.2021.100156","article-title":"Exploring the application and mechanism of sodium hyaluronate in cryopreservation of red blood cells","volume":"12","author":"Liu","year":"2021","journal-title":"Mater. Today Bio"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.cryobiol.2019.11.002","article-title":"Natural cryoprotectants combinations of l-proline and trehalose for red blood cells cryopreservation","volume":"91","author":"Dou","year":"2019","journal-title":"Cryobiology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1021\/bi00866a011","article-title":"Hydrogen ion buffers for biological research","volume":"5","author":"Good","year":"1966","journal-title":"Biochemistry"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1038\/nprot.2006.4","article-title":"Tricine\u2013SDS-PAGE","volume":"1","year":"2006","journal-title":"Nat. Protoc."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"377","DOI":"10.2527\/jas1972.352377x","article-title":"Effect of Some Buffers on Storage and Freezing of Boar Spermatozoa","volume":"35","author":"Crabo","year":"1972","journal-title":"J. Anim. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1016\/0014-5793(86)81230-3","article-title":"Rate constants for reaction of hydroxyl radicals with Tris, Tricine and Hepes buffers","volume":"199","author":"Hicks","year":"1986","journal-title":"FEBS Lett."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1007\/s001340050833","article-title":"An international view of hydroxyethyl starches","volume":"25","author":"Treib","year":"1999","journal-title":"Intensive Care Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1111\/rda.13402","article-title":"Antioxidants and glycine can improve the developmental competence of vitrified\/warmed ovine immature oocytes","volume":"54","author":"Ahmadi","year":"2019","journal-title":"Reprod. Domest. Anim."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1095\/biolreprod.109.081646","article-title":"Rescue of Postcompaction-Stage Mouse Embryo Development from Hypertonicity by Amino Acid Transporter Substrates That May Function as Organic Osmolytes1","volume":"82","author":"Richards","year":"2010","journal-title":"Biol. Reprod."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1093\/humupd\/dmp045","article-title":"Cell volume regulation in oocytes and early embryos: Connecting physiology to successful culture media","volume":"16","author":"Baltz","year":"2010","journal-title":"Hum. Reprod. Update"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"13982","DOI":"10.1073\/pnas.2334537100","article-title":"The glycine neurotransmitter transporter GLYT1 is an organic osmolyte transporter regulating cell volume in cleavage-stage embryos","volume":"100","author":"Hammer","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1007\/s13197-011-0251-1","article-title":"Genesis and development of DPPH method of antioxidant assay","volume":"48","author":"Kedare","year":"2011","journal-title":"J. Food Sci. Technol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1016\/S0308-8146(99)00089-8","article-title":"Applicability of an improved Trolox equivalent antioxidant capacity (TEAC) assay for evaluation of antioxidant capacity measurements of mixtures","volume":"66","author":"Haenen","year":"1999","journal-title":"Food Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.ab.2016.10.021","article-title":"Assessment of lipid peroxidation by measuring malondialdehyde (MDA) and relatives in biological samples: Analytical and biological challenges","volume":"524","author":"Tsikas","year":"2017","journal-title":"Anal. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"13051","DOI":"10.1016\/j.eswa.2012.05.056","article-title":"A state-of the-art survey of TOPSIS applications","volume":"39","author":"Behzadian","year":"2012","journal-title":"Expert Syst. Appl."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1016\/0377-2217(94)90282-8","article-title":"TOPSIS for MODM","volume":"76","author":"Lai","year":"1994","journal-title":"Eur. J. Oper. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1002\/bs.3830220402","article-title":"A theory of analytical hierarchies applied to political candidacy","volume":"22","author":"Saaty","year":"1977","journal-title":"Behav. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"15173","DOI":"10.3390\/ijerph121214976","article-title":"Applying Rank Sum Ratio (RSR) to the Evaluation of Feeding Practices Behaviors, and Its Associations with Infant Health Risk in Rural Lhasa, Tibet","volume":"12","author":"Wang","year":"2015","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ejor.2004.04.028","article-title":"Analytic hierarchy process: An overview of applications","volume":"169","author":"Vaidya","year":"2006","journal-title":"Eur. J. Oper. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1021\/acsmacrolett.0c00044","article-title":"Combinatorial Biomaterials Discovery Strategy to Identify New Macromolecular Cryoprotectants","volume":"9","author":"Stubbs","year":"2020","journal-title":"ACS Macro Lett."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/0022-2836(85)90180-9","article-title":"The active center of catalase","volume":"185","author":"Fita","year":"1985","journal-title":"J. Mol. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1021\/ar50058a001","article-title":"Superoxide radical and superoxide dismutase","volume":"5","author":"Fridovich","year":"1972","journal-title":"Acc. Chem. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1007\/s11738-018-2612-0","article-title":"CAT and MDH improve the germination and alleviate the oxidative stress of cryopreserved Paeonia and Magnolia pollen","volume":"40","author":"Jia","year":"2018","journal-title":"Acta Physiol. Plant."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3089","DOI":"10.20546\/ijcmas.2020.907.364","article-title":"Effects of Ascorbic acid as antioxidant semen additive in cryopreservation of cross-bred cattle bull semen","volume":"9","author":"Singh","year":"2020","journal-title":"Int. J. Curr. Microbiol. App. Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.tmrv.2010.08.004","article-title":"Circulatory Risk in the Transfusion of Red Blood Cells With Impaired Flow Properties Induced by Storage","volume":"25","author":"Barshtein","year":"2011","journal-title":"Transfus. Med. Rev."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1021\/acs.biomac.1c01477","article-title":"Ice Recrystallization Inhibition Is Insufficient to Explain Cryopreservation Abilities of Antifreeze Proteins","volume":"23","author":"Sun","year":"2022","journal-title":"Biomacromolecules"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1309\/YHYL-8XTH-5JM2-43EJ","article-title":"Validation of the Diesse Mini-Ves Erythrocyte Sedimentation Rate (ESR) Analyzer Using the Westergren ESR Method in Patients With Systemic Inflammatory Conditions","volume":"118","author":"Happe","year":"2002","journal-title":"Am. J. Clin. Pathol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1926","DOI":"10.1016\/j.apm.2010.11.001","article-title":"A method for group decision-making based on determining weights of decision makers using TOPSIS","volume":"35","author":"Yue","year":"2011","journal-title":"Appl. Math. Model."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/15\/8462\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:59:52Z","timestamp":1760140792000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/15\/8462"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,30]]},"references-count":48,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["ijms23158462"],"URL":"https:\/\/doi.org\/10.3390\/ijms23158462","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,30]]}}}