{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T11:58:03Z","timestamp":1782993483566,"version":"3.54.5"},"reference-count":31,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2021,10,25]],"date-time":"2021-10-25T00:00:00Z","timestamp":1635120000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000968","name":"American Heart Association","doi-asserted-by":"publisher","award":["19AIREA34450204"],"award-info":[{"award-number":["19AIREA34450204"]}],"id":[{"id":"10.13039\/100000968","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Institute of Health","award":["HL055374"],"award-info":[{"award-number":["HL055374"]}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["1919560"],"award-info":[{"award-number":["1919560"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Ischemic stroke is the most common type of stroke and thrombolytic therapy is the only approved treatment. However, current thrombolytic therapy with tissue plasminogen activator (tPA) is often hampered by the increased risk of hemorrhage. Plasmin, a direct fibrinolytic, has a significantly superior hemostatic safety profile; however, if injected intravenously it becomes rapidly inactivated by anti-plasmin. Nanoformulations have been shown to increase drug stability and half-life and hence could be applied to increase the plasmin therapeutic efficacy. Here in this paper, we report a novel heparin and arginine-based plasmin nanoformulation that exhibits increased plasmin stability and efficacy. In vitro studies revealed significant plasmin stability in the presence of anti-plasmin and efficient fibrinolytic activity. In addition, these particles showed no significant toxicity or oxidative stress effects in human brain microvascular endothelial cells, and no significant blood brain barrier permeability. Further, in a mouse photothrombotic stroke model, plasmin nanoparticles exhibited significant efficacy in reducing stroke volume without overt intracerebral hemorrhage (ICH) compared to free plasmin treatment. The study shows the potential of a plasmin nanoformulation in ischemic stroke therapy.<\/jats:p>","DOI":"10.3390\/ijms222111477","type":"journal-article","created":{"date-parts":[[2021,10,25]],"date-time":"2021-10-25T21:40:21Z","timestamp":1635198021000},"page":"11477","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Heparin and Arginine Based Plasmin Nanoformulation for Ischemic Stroke Therapy"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3736-8899","authenticated-orcid":false,"given":"Ramsha","family":"Aamir","sequence":"first","affiliation":[{"name":"Department of Mechanical Engineering, University of Michigan-Dearborn, Dearborn, MI 48128, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cameron","family":"Fyffe","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering, University of Michigan-Dearborn, Dearborn, MI 48128, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7244-889X","authenticated-orcid":false,"given":"Netanel","family":"Korin","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Technion-Israel Institute of Technology, Haifa 32000, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel A.","family":"Lawrence","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enming J.","family":"Su","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathumai","family":"Kanapathipillai","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering, University of Michigan-Dearborn, Dearborn, MI 48128, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,25]]},"reference":[{"key":"ref_1","first-page":"e29","article-title":"Heart disease and stroke statistics--2015 update: A report from the American Heart Association","volume":"131","author":"Mozaffarian","year":"2015","journal-title":"Circulation"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1016\/S1474-4422(12)70104-7","article-title":"Intracerebral haemorrhage: Mechanisms of injury and therapeutic targets","volume":"11","author":"Keep","year":"2012","journal-title":"Lancet Neurol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"The NINDS t-PA Stroke Study Group (1997). Intracerebral hemorrhage after intravenous t-PA therapy for ischemic stroke. Stroke, 28, 2109\u20132118.","DOI":"10.1161\/01.STR.28.11.2109"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1679","DOI":"10.1161\/01.CIR.0000012747.53592.6A","article-title":"Markers of increased risk of intracerebral hemorrhage after intravenous recombinant tissue plasminogen activator therapy for acute ischemic stroke in clinical practice: The Multicenter rt-PA Stroke Survey","volume":"105","author":"Tanne","year":"2002","journal-title":"Circulation"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1016\/S0140-6736(04)15692-4","article-title":"Association of outcome with early stroke treatment: Pooled analysis of ATLANTIS, ECASS, and NINDS rt-PA stroke trials","volume":"363","author":"Hacke","year":"2004","journal-title":"Lancet"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1503\/cmaj.1041561","article-title":"Thrombolysis for acute ischemic stroke: Results of the Canadian Alteplase for Stroke Effectiveness Study","volume":"172","author":"Hill","year":"2005","journal-title":"CMAJ"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1159\/000103110","article-title":"Symptomatic intracerebral hemorrhage following thrombolytic therapy for acute ischemic stroke: A review of the risk factors","volume":"24","author":"Lansberg","year":"2007","journal-title":"Cerebrovasc. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.1001\/jama.2015.3058","article-title":"Acute stroke intervention: A systematic review","volume":"313","author":"Prabhakaran","year":"2015","journal-title":"JAMA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"S129","DOI":"10.1161\/STROKEAHA.113.001457","article-title":"Temporal trends in acute stroke management","volume":"44","author":"Kleindorfer","year":"2013","journal-title":"Stroke"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"National Institute of Neurological Disorders and Stroke rt-PA Stroke Study Group (1995). Tissue plasminogen activator for acute ischemic stroke. N. Engl. J. Med., 333, 1581\u20131587.","DOI":"10.1056\/NEJM199512143332401"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"866","DOI":"10.1016\/S1474-4422(10)70165-4","article-title":"Implementation and outcome of thrombolysis with alteplase 3-4.5 h after an acute stroke: An updated analysis from SITS-ISTR","volume":"9","author":"Ahmed","year":"2010","journal-title":"Lancet Neurol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2352","DOI":"10.1016\/S0140-6736(12)60768-5","article-title":"The benefits and harms of intravenous thrombolysis with recombinant tissue plasminogen activator within 6 h of acute ischaemic stroke (the third international stroke trial [IST-3]): A randomised controlled trial","volume":"379","author":"Sandercock","year":"2012","journal-title":"Lancet"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3034","DOI":"10.1161\/01.STR.31.12.3034","article-title":"Metalloproteinase inhibition reduces thrombolytic (tissue plasminogen activator)-induced hemorrhage after thromboembolic stroke","volume":"31","author":"Lapchak","year":"2000","journal-title":"Stroke"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1161\/STROKEAHA.111.635250","article-title":"A rat model of studying tissue-type plasminogen activator thrombolysis in ischemic stroke with diabetes","volume":"43","author":"Fan","year":"2012","journal-title":"Stroke"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1038\/nm1787","article-title":"Activation of PDGF-CC by tissue plasminogen activator impairs blood-brain barrier integrity during ischemic stroke","volume":"14","author":"Su","year":"2008","journal-title":"Nat. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1007\/s00401-017-1749-z","article-title":"Microglial-mediated PDGF-CC activation increases cerebrovascular permeability during ischemic stroke","volume":"134","author":"Su","year":"2017","journal-title":"Acta Neuropathol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"S56","DOI":"10.1016\/S0049-3848(09)70145-8","article-title":"Thrombolytic therapy for deep vein thrombosis: Potential application of plasmin","volume":"123","author":"Marder","year":"2009","journal-title":"Thromb. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1111\/j.1538-7836.2009.03701.x","article-title":"Direct fibrinolytic agents: Biochemical attributes, preclinical foundation and clinical potential","volume":"8","author":"Marder","year":"2010","journal-title":"J. Thromb. Haemost."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1034","DOI":"10.1046\/j.1538-7836.2003.00128.x","article-title":"Thrombolytic potency of acid-stabilized plasmin: Superiority over tissue-type plasminogen activator in an in vitro model of catheter-assisted thrombolysis","volume":"1","author":"Novokhatny","year":"2003","journal-title":"J. Thromb. Haemost."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1007\/s12975-014-0376-4","article-title":"Plasmin-loaded echogenic liposomes for ultrasound-mediated thrombolysis","volume":"6","author":"Kandadai","year":"2015","journal-title":"Transl. Stroke Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1016\/j.nano.2017.12.022","article-title":"A targeted ferritin-microplasmin based thrombolytic nanocage selectively dissolves blood clots","volume":"14","author":"Seo","year":"2018","journal-title":"Nanomedicine"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1038\/nrd2591","article-title":"Strategies in the design of nanoparticles for therapeutic applications","volume":"9","author":"Petros","year":"2010","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2971","DOI":"10.1039\/c2cs15344k","article-title":"Targeted polymeric therapeutic nanoparticles: Design, development and clinical translation","volume":"41","author":"Kamaly","year":"2012","journal-title":"Chem. Soc. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2004","DOI":"10.1111\/jth.14637","article-title":"Engineered microparticles and nanoparticles for fibrinolysis","volume":"17","author":"Disharoon","year":"2019","journal-title":"J. Thromb. Haemost."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1126\/science.1217815","article-title":"Shear-activated nanotherapeutics for drug targeting to obstructed blood vessels","volume":"337","author":"Korin","year":"2012","journal-title":"Science"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1002\/wnan.68","article-title":"Heparin-based nanoparticles","volume":"2","author":"Kemp","year":"2010","journal-title":"Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol."},{"key":"ref_27","first-page":"1872","article-title":"Significant role of cationic polymers in drug delivery systems","volume":"46","author":"Farshbaf","year":"2018","journal-title":"Artif. Cells Nanomed. Biotechnol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1038\/246355a0","article-title":"Anticoagulant action of heparin","volume":"246","author":"Damus","year":"1973","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/S0049-3848(01)00368-1","article-title":"Arginine inhibits hemostasis activation","volume":"104","author":"Stief","year":"2001","journal-title":"Thromb. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4752","DOI":"10.1182\/blood-2011-07-369512","article-title":"t-PA-specific modulation of a human blood-brain barrier model involves plasmin-mediated activation of the Rho kinase pathway in astrocytes","volume":"119","author":"Niego","year":"2012","journal-title":"Blood"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Niego, B., Lee, N., Larsson, P., De Silva, T.M., Au, A.E., McCutcheon, F., and Medcalf, R.L. (2017). Selective inhibition of brain endothelial Rho-kinase-2 provides optimal protection of an in vitro blood-brain barrier from tissue-type plasminogen activator and plasmin. 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