Importantly, salmon FG was the most resistant to degradation by cellular proteases. collectively, blood immunological and coagulation guidelines support the suitability of salmon-derived FG in the development of fibrin sealants for medical use. Keywords:fibrin glue, coagulation, thrombin time, antibodies, animal experiments == Intro == Fibrin glues (FG) of thrombin and fibrinogen composition were originally used to minimize blood loss during procedures in medical practice.1Current use and studies have expanded to protect many areas in biomedicine and reveal the potential of FG in cell therapy and tissue executive.26Studies have been conducted to assess FG like a potential controlled or extended launch system for medicines. Fibrin, as a relatively slowly degrading biological scaffold can be loaded with active ingredients to accomplish high local concentration of medicines for controlled launch.712However, the main use and experimental study is still focused on haemostasis and wound healing in a variety of surgical settings. FGs currently authorized for medical use are derived PEG3-O-CH2COOH from human being or bovine plasma. As any plasma-derived product these materials present potential risk for transmission of bacteria, viruses, or prions. This concern limits widespread use of mammalian plasma-derived FG. Use of bovine proteins in FG may also compromise the patient homeostasis. Development of antibodies and their mix reactions to sponsor coagulation proteins can seriously alter normal haemostasis. For example, two thirds of reports about the event of antibodies against coagulation element V are due to earlier bovine thrombin exposure.13 Our earlier studies showed that Atlantic salmon blood proteins represent a potentially safer, equally effective, and less costly alternative to human being or additional mammalian blood proteins. Coagulation and immunologic studies inside a pilot study on rats and rabbits, treated intraperitoneally with salmon FG, showed no deleterious effects on coagulation profiles and no cross-reactivity with sponsor fibrinogen or thrombin.14Similarly, Ju et al. assessed the growth of mammalian neurons in bovine, human being, and salmon fibrin and found that salmon fibrin gels motivated neurite (dendrite and axon) growth to the PEG3-O-CH2COOH greatest degree. Importantly, salmon FG was the most resistant to degradation by cellular proteases. It was suggested that salmon FG may be a beneficial scaffold for neuronal regrowth after central nervous system injury. 15 In this study, we lengthen our earlier experimental investigations aiming to assess the peripheral blood coagulation profiles and antibodies against FG parts in enzyme-linked immunosorbent and immunoblot assays on rats intraperitoneally treated with PEG3-O-CH2COOH salmon FG. We also analyzed possible cross-reactivity to human being element Va and identified C reactive protein levels to assess acute phase response before and after the intraperitoneal administration of salmon FG. == MATERIALS AND METHODS == == Coagulation proteins == Salmon fibrinogen PEG3-O-CH2COOH (Lot #1289) and thrombin (Lot #5031), purified as previously explained were used for administration to experimental animals and as antigen in immunologic studies. Salmon fibrinogen was treated with 10 kGy gamma irradiation. Human being Element Va (Lot L6050564 C6050564) was from US Biologicals (Massachusetts, MA). == Secondary antibodies, enzyme substrates, along with other materials == Rabbit alkaline phosphatase-conjugated antirat IgG (A6066, Sigma, St. Louis) was used in combination with the enzyme substrates p-nitrophenyl phosphate (PNPP, Sigma, St. Louis, USA) at a concentration of 1 1 mg/ml in diethanolamine buffer, pH-9.8, in ELISA and 5-bromo-4-chloro-3-indolyl PEG3-O-CH2COOH phosphate/nitroblue tetrazolium (BCIP/NBT) (B-8503 Lot 32K1430; N-6876 Lot 32K5307, Sigma-Aldrich, St. Louis, MO) in immunoblot assays. Microtiter Assembly Strip 18 enhanced binding, EB ELISA strip-plates (Cat no. 95029180, ThermoElectron OY, Finland) were used in ELISA and Trans-Blot Transfer Medium Pure Nitrocellulose Membrane (NCM) at 0.2 m (Control 13108, Bio-Rad Laboratories, CA) in immunoblotting throughout the studies. Swine serum was separated from clotted swine blood obtained from a local slaughterhouse by centrifugation. == Experimental animals == Forty-six Wistar KLRC1 antibody rats (24 males, 22 females) were bred and kept at the local animal care facility (Biomedicum’s Animal House, University or college of Tartu, Estonia). Rats were 11 months aged, weighting 280418 g (females) and 450748 g (males) at the start of.