The effect of ethyl acetate mediated silver nanoparticles from Urtica diocia on hemostasis; in-vitro and in-vivo study

Abstract

Bleeding may easily result in death during surgery or military operations. The need for safer and more effective hemostatic materials has received wide attention. Most recent topical hemostatic agents were demonstrated to be ineffective at reducing bone bleeding. Searching among biosynthesized nanoparticles (NPs) has provided promising results. This study reports an ethyl acetate-mediated silver nanoparticle (AgNP) effect using Urtica diocia (UD) leaves for bone hemostasis. The active components of UD-AgNPs were identified by gas chromatography–mass spectrometry (GC‒MS). MTT assays were used to evaluate the cytotoxicity of UD-AgNPs. Hemorrhagic circular defects (3 mm) were created on rat femur bone (n = 40). Group I: defect left unfilled (negative control), and the remaining defects were filled with UD-AgNPs, bone wax or Surgicel in groups II, III and IV, respectively. The bleeding times and amounts were calculated for each group. Prothrombin time (PT), activated partial thromboplastin time (aPTT), platelets (PLT), white blood cells (WBCs), red blood cells (RBCs) and hemoglobin (Hgb) were measured at baseline and one and four weeks postoperatively. GC‒MS identified the active components involved in bleeding control, bone regeneration and antibacterial activities, such as phytol, neophytadiene, carboxylic acid and hexadecenoic acid. MTT assays showed cell viabilities greater than 75% for all UD-AgNP concentrations against normal cells. UD-AgNPs demonstrated immediate hemostasis (7.7 s), similar to bone wax (7.3 s), with a similar amount of blood loss (0.05 gm). UD-AgNPs increased PLT (1347.5 × 103/μL) and reduced aPTT (12.6 s) and maintained normal WBCs compared to bone wax (19.2 × 103/μL) at the first week. These observations suggested that UD-AgNPs are an effective hemostatic agent with good biocompatibility.