In Situ Microparticles Loaded with S-Nitrosoglutathione Protect from Stroke

PLoS One. 2015 Dec 8;10(12):e0144659. doi: 10.1371/journal.pone.0144659. eCollection 2015.

Abstract

Treatment of stroke, especially during the first hours or days, is still lacking. S-nitrosoglutathione (GSNO), a cerebroprotective agent with short life time, may help if administered early with a sustain delivery while avoiding intensive reduction in blood pressure. We developed in situ forming implants (biocompatible biodegradable copolymer) and microparticles (same polymer and solvent emulsified with an external oily phase) of GSNO to lengthen its effects and allow cerebroprotection after a single subcutaneous administration to Wistar rats. Arterial pressure was recorded for 3 days (telemetry, n = 14), whole-blood platelet aggregation up to 13 days (aggregometry, n = 58), and neurological score, cerebral infarct size and edema volume for 2 days after obstruction of the middle cerebral artery by autologous blood clots (n = 30). GSNO-loaded formulations (30 mg/kg) induced a slighter and longer hypotension (-10 vs. -56 ± 6 mmHg mean arterial pressure, 18 h vs. 40 min) than free GSNO at the same dose. The change in pulse pressure (-50%) lasted even up to 42 h for microparticles. GSNO-loaded formulations (30 mg/kg) prevented the transient 24 h hyper-aggregability observed with free GSNO and 7.5 mg/kg-loaded formulations. When injected 2 h after stroke, GSNO-loaded microparticles (30 mg/kg) reduced neurological score at 24 (-62%) and 48 h (-75%) vs. empty microparticles and free GSNO 7.5 mg/kg and, compared to free GSNO, divided infarct size by 10 and edema volume by 8 at 48 h. Corresponding implants reduced infarct size and edema volume by 2.5 to 3 times. The longer (at least 2 days) but slight effects on arterial pressures show sustained delivery of GSNO-loaded formulations (30 mg/kg), which prevent transient platelet hyper-responsiveness and afford cerebroprotection against the consequences of stroke. In conclusion, in situ GSNO-loaded formulations are promising candidates for the treatment of stroke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Pressure / drug effects
  • Disease Models, Animal
  • Injections, Subcutaneous
  • Male
  • Microspheres
  • Neuroprotective Agents / administration & dosage
  • Neuroprotective Agents / therapeutic use*
  • Platelet Aggregation / drug effects
  • Rats
  • Rats, Wistar
  • S-Nitrosoglutathione / administration & dosage
  • S-Nitrosoglutathione / therapeutic use*
  • Stroke / drug therapy*
  • Telemetry

Substances

  • Neuroprotective Agents
  • S-Nitrosoglutathione

Grants and funding

This work was supported by the Agence Nationale de la Recherche [ANR NanoSNO project 2010–2014, managed by P. LEROY, Université de Lorraine, EA3452 CITHEFOR]; and the French Ministry of Research and Education (Paris, France). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. There is no commercial affiliation between ETAP-Lab and the other laboratory implicated in this work. ETAP-Lab did not provide any financial support to the present project. ETAP-Lab provided salaries only for authors that are members of its staff: Nicolas Violle and Jean-François Bisson. ETAP-Lab realized the stroke study in its own facilities on the demand of EA3542. For this study, EA3542 provided GSNO, formulations and consumable materials (Animals, syringe,…). ETAP-Lab provided surgeons, experimenters and laboratory technician, as well as facilities and equipment. ETAP-Lab participated to the study design, data collection and analysis, decision to publish, and preparation of the manuscript relatives to the stroke study.