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Abstract Infectious diseases represent a great risk as more than half of the deaths happening worldwide. Brucellosis is one of the diseases that caused by Brucella species, a bacterial pathogen. Despite the profound success achieved by the usage of antibiotics against infectious diseases, bacterial infections still conseder one the greatest challenges the in global healthcare. In the last few years, nanotechnology allows the development of new products that are used in human medicine; for quick and specific drug delivery into cells and tissues. The present study was aimed at preparing and characterizeing the nano- albumin particles and evaluateing thier safety on living lymphocyte cell culture for their potential therapeutic as drug carrier to enhance the treatment of brucella infection in male Balb.c mice. The study was divided in to three parts, the first part aimed to prepare the nano-albumin particles using the desolvation technique. Three techniques were used for characterization of the prepared particles; Transmission Electron Microscopy (TEM) was used to determine the particles shape and size; Zeta Sizer to estimate the zeta potential of the surface of the prepared nano particles and the size of this particles and finally by using X Ray Diffraction (XRD) technique to assess the chemical structure of the prepared nanoparticles. The second part aimed to evaluate the safety of the prepared nano-albumin In Vitro by applying MTT test for the cytotoxicity, comet test for genotoxicity and also study of immune effect of this particles on living lymphocyte cells. Finally, the third part amid to load two different types of antibiotics (Streptomycin and Doxycycline) as combination therapy and (Gentamycin) as mono 111 therapy on three different concentrations of the prepared nano-albumin and to follow its efficiency to treat the intracellular Brucella infection. This was by infecting male Balb/c mice intraperitoneally with 105 CFU of B. melitensis 16M and divideding the mice into two groups, The first group was diveded to six groups: negative group (received only saline), positive gp. (infected with brucella without treatment), 3ed gp.received two doses of non loaded form (free form) of (180 µg Streptomycin and 36 µg Doxycycline) / mouse/ dose intraperitoneally, 4th, 5th and 6th gps. received the loaded form of (180 µg streptomycin and 36 µg doxycycline) / mouse/ douse loaded on three different concentration of nano-albumin (1mg/ml, 0.5 mg/ml, 0.25 mg/ml). As a combination therapy, the second group also divided to six group: negative group (received only saline), positive gp. (infected with brucella without treatment), 3ed gp. received three doses of nonloaded form of 100µg gentamycin / mouse/ dose and 4th, 5th and 6th gps received the loaded form of 100µg Gentamycin / mouse/ dose on three different concentration of nano- albumin (1mg/ml, 0.5 mg/ml, 0.25 mg/ml) as mono therapy, three mice were sacarified after each dose and after 5 days of last dose in combination therapy and after one week of last dose of mono therapy to collect the spleen that used to follow the treatment by molecular study using PCR and histopathology studies. The prepared nano-albumin particles were nearly spherical in shape and haveing smooth surface as determined by Transmission Electron Microscopy (TEM). The sizes of the obtained nanoparticles were 70 ± 10 nm with negative surface zeta potential. Additionally, the in vitro safety of albumin nanoparticles has been demonstrated. Both cytotoxicity and genotoxicity studies indicated that, there was no observed toxic effect of nano-albumin on lymphocytes. Also, the results 111 showed that the albumin nanoparticles enhances and promotes the response of lymphocytes to PHA miotagen. The produced nano- albumin particles could succefully interact with streptomycin, doxaciclin and gentamicin to be used as a drug delivary system. Also the results showed that by using the different forms of antibiotics (combination therapy and mono therapy) loaded on three different concentrations of nano- albumin (1mg/ml, 0.5 mg/ml, 0.25 mg/ml ), the loaded form of the two types of therapy was more effective in the treatment of the infection with B. melitensis than the non loaded forms. The two concentrations (0.5 mg/ml and 0.25 mg/ml) were more effective than the high concentration (1mg/ml) since the two low concentrations cleared the infection totally (100%) but at the high concentration it decreased the infection and the treatment in (S+D) to 66% and in (G) was 33%. Finally the combination therapy was more effective than the mono therapy in the intracellular infection treatment. In conclusion, the nano- albumin is easy to prepare, have no toxic effect on living cells and it enhance the immune proliferation activity of lymphocyts. Also it could successfully interact with different drugs. So the of nano- albumin can be used as a carrier for antibiotic for treatment of intracellular infection like Brucella. Further studies are also required to confirm these preliminary results. Moreover, following up the treatment efficiency for longer time are required. |