Effect of Using Zinc Oxide and Gold Nanoparticles on ...

Republic of Iraq Ministry of Higher Education &Scientific Research University of Baghdad/College of Science Biotechnology Department

Effect of Using Zinc Oxide and Gold Nanoparticles on Experimentally Induced Diabetic Rats

A thesis Submitted to the College of Sciences at the University of Baghdad In Partial fulfillment of the Requirements for the Degree of Doctor of Philosophy of Sciences in Biotechnology By

Laith Ahmad Yaaqoob M.Sc.: Genetic Engineering and Biotechnology Institute for post graduate / University of Baghdad in 2005 B.Sc.: Plant Protection Department / Agriculture Engineering College / University of Baghdad in 1999

Supervised by

Prof. Dr. Hasan F. Samir

2016 A.D.

1437 A.H.

‫ْ‬ ‫ْ‬ ‫َٰ‬ ‫ِبس ِم اللَّ ِه ال َّرحمن ال َّ ِرح مي‬ ‫ِ‬ ‫ك َْ‬ ‫تَ َب َار َك ذ ِاَّلي ِب َي ِد ِه الْ ُم ْ ُ‬ ‫ْل َوه َُو عَ َ َٰل ُ ِ ل‬ ‫َش ٍء‬ ‫قَ ِدير ﴿‪﴾1‬‬

‫ل‬ ‫ع‬ ‫صدق هللا ا ظيم‬ ‫سورة تبارك‬

Certification

Certification I certify that this thesis was prepared under my supervision at the department of Biotechnology, College of sciences, and University of Baghdad as a partial requirement for the Degree of Doctor of Philosophy in Biotechnology.

Signature: Supervisor prof. Dr. Hassan F. Samir Department of Biotechnology College of Sciences University of Baghdad

In view of available recommendation, I forward this thesis debate by the Examination Committee

Signature: Prof. Dr. Muayad Sabri Shawkat Department head of Biotechnology College of Sciences University of Baghdad

Committee Certification

Committee Certification We certify that we have read this thesis, entitled "Effect of Using Zinc Oxide and Gold Nanoparticles on Experimentally Induced Diabetic Rats", and as Examining Committee, examined the student Laith Ahmad Yaaqoob on its content, and that in our opinion it is adequate for the partial fulfillment of the requirements for the Degree of Doctor of Philosophy in Biotechnology.

Signature: Name: Dr. Muayad Sabri Shawkat Title: Professor Date: / / (Chairman)

Signature: Name: Dr. Salim Rasheed Hamoudi Alobaidie Title: Professor Date: / / (Member)

Signature: Name: Dr. Salwa H. N. Al-Rubae'i Title: Professor Date: / / (Member)

Signature: Name: Dr. Abas Mahdi Rhma Title: Assistant Professor Date: / / (Member)

Signature: Name: Dr. Muhammed Mizher Radhi Title: Assistant Professor Date: / / (Member)

Signature: Name: Dr. Hasan F. Al-Azzawie Title: Professor Date: / / (Supervisor)

Approved by the Dean of the college of Science. Signature: Name: Dr. Fadhil Abd Rasin Title: Assistant Professor Date: / / Dean of the College of Science / University of Baghdad

Dedication

Dedication This thesis is dedicated to my parents, my

wife, my brothers and my children My endless love for them to support and encouragement

Laith

Acknowledgements

Acknowledgements At the beginning, I would like to thank my gratitude to Allah merciful for complete this research and for helping me and giving the patience along the life. I would like to express my thanks and full appreciation and deep gratitude to my supervisor Professor Dr.Hassan F. Samir Al-azzawie, for his advice, guidance, and invaluable suggestions and endless support throughout the entire period of my work. He has been a constant source of inspiration and encouragement. I would like to express my special thanks to Assistant professor Dr.Salah Mahdy in Al-Nahrain Research Center for his supervision and guidance in the animal experimental work and for his kind helpfulness. I am also most grateful to the head of Biotechnology Department professor Dr. Muayad Sabri Shawkat, I am very grateful and would like to express my sincere appreciation to the members of biotechnology department, College of sciences, Baghdad University for their kind assistance during the work. Last but not the least, I express my deep gratitude to my family and thanks to all my colleagues and my friends for their continuous interest to complete this work, Thanks also go to everyone who helps me..

Summary

Summary This study developed a rapid, ecofriendly and convenient green environment method for the synthesis of stable Zinc oxide nanoparticles (ZnO NPs) of average diameter of 40 ± 5.0 nm and gold nanoparticles (Au NPs) of average diameter of 35 ± 5.0 using the aqueous solution of Olea europeae leaves extract as reducing and capping agent used to increase the stability of the nanoparticles. The characterization studied was done by UV-Visible spectroscopy, Scanning electron microscopy (SEM), Atomic force microscopy (AFM), X-Ray diffraction (XRD), Fourier Transmission infrared spectroscopy (FTIR)

Application of nanoparticles in medicine is an attractive proposition. In the present study, the antihyperglycemic effect of intake each gold, and ZnO NPs alone and their mixture at different concentrations on oxidative stress status in alloxan induced diabetes rats were evaluated. One hundred and thirty male albino rats with weight 320 ± 25 grams and age 6 months were used in experimental design. Animals were grouped randomly to thirteenth group (each group=10) as follows: first group as healthy non diabetic control (G1) did not receive any type of treatment. Second group as healthy non diabetic control receive either of Au and ZnO NPs or their mixture according to the experimental design (G2).The other groups treated intraperitoneally once with 150 mg/kg b.w of alloxan, then when these rats became hyperglycemic, they divided to eleven groups .Group (G3) as diabetic group did not receive any type of treatment. Groups (G4, G5 and G6) as Diabetic rats received intraperitoneally single daily dose of (2.5, 5.0 and 10) mg/kg b.w ZnO NPs in suspension respectively for 8 weeks. Diabetic rats received a single daily subcutaneous dose of mixed I

Summary

insulin 2 U/kg b.w (G7). Diabetic rats received intraperitoneally single daily dose of (2.5, 5.0 and 10) mg/kg b.w Au NPs in suspension respectively (G8, G9 and G10), and finally diabetic rats received intraperitoneally single daily dose of half concentration of mixture of both nanoparticles (G11, G12 and G13). At the end of experimental time 8 weeks, fasting blood glucose, serum insulin, glycoslated HbA1c, malondialdehyde (MDA), reduced glutathione (GSH) and serum activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and Catalase (Cat), in addition lipid profile were determined in all rat groups. The results indicated that the blood glucose, MDA levels were increased highly significantally (P