Bio-nanomaterials for Biomedical Technology

Bio-nanomaterials for Biomedical Technology

Bio-nanomaterials for Biomedical Technology

Editors V. Rajendran P. Prabu K.E. Geckeler

© KSR Group of Institutions, 2015 First Published, 2015 All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage or retrieval system, without prior permission in writing from the copyright holder. No responsibility for loss caused to any individual or organization acting on or refraining from action as a result of the material in this publication can be accepted by Bloomsbury India or the author/editor. BLOOMSBURY PUBLISHING INDIA PVT. LTD. New Delhi London Oxford New York Sydney ISBN: 978-93-85436-92-5 10 9 8 7 6 5 4 3 2 1 Published by Bloomsbury Publishing India Pvt. Ltd. DDA Complex LSC, Building No. 4, 2nd Floor Pocket 6 & 7, Sector – C Vasant Kunj, New Delhi 110070 Printed at anvi composers, Paschim Vihar, New Delhi

The publisher believes that the contents of this book do not violate any existing copyright/intellectual property of others in any manner whatsoever. However, in case any source has not been duly attributed, the publisher may be notified in writing for necessary action.

Foreword The past few decades have seen unprecedented advancements in the field of nanotechnology with spectacular developments in a wide area of Material Science. Currently, nanotechnology is a fast growing field with a broad spectrum of applications and it is appreciated that the Centre for Nano Science and Technology (CNST) of the K.S. Rangasamy College of Technology (KSRCT) has undertaken an initiative to organize an International Conference on Nanomaterials and Nanotechnology (NANO-15) with a special topic on ‘Research to Innovations to Technology Transfer’ in India, especially at Tamil Nadu. Based on its good infrastructure and human resources, CNST develops R & D activities with international standards by many funded projects and research publications. KSRCT collaborates with academic institutions and national and international research laboratories/ industries of high reputation. The conference features many plenary and key note addresses presented by invited speakers and more than 1100 delegates from around the world are participating, interacting, and discussing the exciting and rapidly developing aspects of Nano Science and Technology. I trust that this conference will be an ideal platform for the presentation and discussion of new concepts and developments of new functional nanomaterials and their applications in new devices and sensors. NANO-15 provides a forum to discuss eco-friendly technologies and promote interactions and collaborations between the delegates. I appreciate that the organising team publishes peer reviewed papers in six independent books. The articles therein will describe new ideas in a rapidly developing field and so stimulate further progress. I am pleased to write a foreword for these books to be published during NANO-15 and wish the Conference participants a fruitful and enjoyable stay in India and I want to thank the organizing team for the their kind helpfulness and hospitality. 20.11.15

Dr. H.C. Mult Robert Huber FMRS Noble Laureate

Message MeSSaGe

Date : 21.11.2015

I am happy to note that our Centre for Nano Science and Technology (CNST) of K. S. Rangasamy College of Technology (KSRCT) is organising an International Conference on Nanomaterials and Nanotechnology (NANO-15) with a special topic on Research to Innovations to Technology transfer during December 07-10, 2015 at our campus. The CNST established with good infrastructure facilities to meet the Scientists and Academicians to an advanced level in the field of nanotechnology. It also focuses in organising such International conferences and workshops to recognize the research outcomes of the young researchers. NANO-15 is organized in KSRCT with plenary lectures by Noble laureates and distinguished scientists, Key note address, invited talks and more than 550 contributed papers. The plenary talk by Nobel Laureates and invited talks from reputed organizations of India and abroad would bring out the current status in material science and technology. I ensure that the participants will have effective deliberations through this conference. I thank Dr. K. Thyagarajah, Principal, KSRCT and Dr V. Rajendran, Director R&D, Organising Chair and his team to organise this event as a successful. The kind support from the various government and private organisations/agencies for the successful conduct of the conference is highly acknowledged. I extend my warm greetings to all the participants and best wishes for the success of the Conference.

Dr. K. S. Rangasamy MJF

Sponsors Science and Engineering Research Board, Department of Science and Technology, New Delhi

Defence Research and Development Organisation, New Delhi

Board of Research in Nuclear Sciences, Mumbai

Indian Council for Medical Research, New Delhi

Tamilnadu State Council for Science and Technology, Chennai

Indian Society for Technical Education, New Delhi

Axis Bank Limited, India

Co-Sponsors National Institute for Nanotechnology (NINT) Innovation Centre, Alberta, Canada NanoCanada, Canada

Silver Sponsor Shimadzu India Pvt. Ltd, Chennai

x Sponsors

The Professor Venkatachalam Rajendran Research Foundation

Exhibitors

CSIR - Central Glass and Ceramics Research Institute, Kolkata

Tekna Plasma India, Chennai, India Lark Innovative Fine Teknowledge, Chennai

Industrial Partners Exigo Knowledge Ventures Private Limited, Bangaluru

Global Connect Inc., Saskatoon, Canada

Higginbothams Private Limited, Chennai, Tamil Nadu

Samraj Constructions, Tamil Nadu

Talent2Success Learning Pvt. Ltd, India Zealtech Electromec India Private Limited, Tamil Nadu

Sponsors xi

Publication Partners Bloomsbury Publishing India Pvt. Ltd, New Delhi The Higher Education Review, Bangalore

Journal Partners Polymer International

Nano System : Physics, Chemistry, Mathematics

Synthesis and Reactivity in Inorganic, Metal Organic and Nano-Metal Chemistry IET Nanobiotechnology

Media Partner The Hindu

Hospitality Partner Radisson 5 Star Hotel, Salem

xiv International Organising Committee

Editors’ Profile Dr. V. Rajendran FUSI, FASI, FInstP, is Director, Research & Development, K S R Group of Institutions and Centre for Nano Science and Technology, K.S Rangasamy College of Technology, Tamil Nadu, India. Under his able guidance, 20 scholars have completed and 09 scholars are pursuing their Ph.D. degrees. He has published more than 200 research papers in reputed international and national journals, 60 papers in conference proceedings, 32 refereed books, 4 R&D books and 11 patents. He has won many awards including the UNESCO visiting Scientist Fellowship, South Africa (2016), Fulbright Fellowship (2015), USA, PSN National Award for Excellence in Science (2013), Prof. K. Arumugam National Award in 2011, Best faculty award in 2010, Raman-Chandra Sekhar silver medal (2010), Tamil Nadu Scientist Award, NDT Man of Year 2004, Indo –Australia Senior Scientist Science and Technology visiting fellowship (2013), DAAD from Germany (2002), INSA, TNSCST Young Scientist, DAE/ BRNS Visiting Scientist, Best paper award from MRSI, ASI, ASA and USI and Outstanding Organiser Award for the 7th National Symposium on Ultrasonics, 1996. Dr. P. Prabu is an Associate Professor in the Department of Nanoscience and Technology. He has two years experience as a post doctorate in Chonbuk National University in the Republic of Korea and also in Nanyang Technological University, Singapore. He obtained his Ph.D. degree from Chonbuk National University, Republic of Korea. He has to his credit more than 23 research papers in reputed international journals, 3 books to his credit. He also attended many international conferences in various countries like South Korea, Thailand, Singapore, Taiwan etc. He has professional affiliation with Korea Tissue Engineering and Regenerative Medicine Society and also member of the Korean Polymer Society and Korean Fiber Society from 2005 till present. Dr. K.E. Geckeler is affiliated with the Gwangju Institute of Science and Technology (GIST), South Korea. and is a Professor at the School of Materials Science and Engineering. He has been the Founding Chair of the Department of Nanobio Materials and Electronics, World Class University (WCU), Gwangju, South Korea. In addition, he serves as Vice Director of the Gruenberg Center for Magnetic Nanomaterials (GCMN). He received his Ph.D. and M.D. degrees from the University of Tuebingen, Germany (both degrees: “magna cum laude”) and spent sabbatical leaves at Harvard University, University of Montana, Clemson University (USA), and at the University of Montpellier (France). He received a series of prestigious awards including the “Fonds of the Chemical Industry”, the “Fritz-Ter-Meer Award”, and the “Science Prize of the President of Korea”. The biannual international IUPAC symposium series on “Macro- and Supra-molecular Architectures and Materials (MAM)” has been initiated and co-organized by him. Prof. Geckeler is Editor-in-Chief of the journal "Polymer International", published by John Wiley & Sons, and is also on editorial boards of a series of other international journals. He has published more than 350 research articles and short communications, 12 book chapters, 15 books, and over 130 patents. His recent books cover different aspects of nanomaterials including the two standard references: “Advanced Nanomaterials” and “Functional Nanomaterials” published by Wiley.

Preface The International Conference on Nanomaterials and Nanotechnology (NANO-15), third in this series with a special topic on Research to Innovation to Technology transfer is organised by Centre for Nano Science and Technology (CNST) of K. S. Rangasamy College of Technology, Tamil Nadu, India during December 07-10, 2015. CNST has established the state-of-the-art experimental facilities, offering undergraduate (B.Tech.), post-graduate (M.Tech.) and research (Ph.D.) programmes and looking for industrial collaboration and partners for the need based development of products in nanotechnology. NANO-15 has been received tremendous supports and overwhelming responses worldwide. More than 950 abstracts have been received from 33 countries. All the received papers are classified under six titles namely Synthesis and Fabrication of Nanomaterials, Advanced Nanomaterials: Synthesis and Applications, Nanoelectronics and Sensors, Applications of Nanostructured Materials for Energy and Environmental Technology, Bio-nanomaterials for Biomedical Technology and Industrial Applications of Nanostructural Materials. 450 have been selected from 600 received full papers, and peer reviewed by the expert committee for the publications in conference books. Out of total 450 full papers accepted for NANO-15, a total of 61 have been identified for the inclusion in the book entitled Bio-nanomaterials for Biomedical Technology after peer review. This book is a collective of biology, chemistry, materials science, engineering and nanotechnology to present an interdisciplinary approach for solving of problems in biomedical technology. The effort put forth by the organizing team in getting the review of all the contributed papers is commendable. All the contributed authors are extended by our sincere thanks for their timely submission and cooperation in carrying out suggestions by the referees. We owe our special thanks to members of technical committee for peer review of contributed papers. The various government funding agencies, private organizations and industries are thankful for their munificent support and sponsor for the successful conduct the conference. The various committee chairs and members are highly acknowledged for making this event a grant success. The support extended by Bloomsbury Publishing India Pvt. Ltd in bringing out this book on time is highly appreciated. V. Rajendran P. Prabu Kurt E. Geckeler

Contents Foreword v Message vii Sponsors ix International Organising Committee xiii Editors’ Profile xv Preface xvii

Plenary Speakers 1. Structural Aspects of Protease Control in Health and Disease Robert Huber

Keynote Speakers 2. Nanoparticles for Bio-Imaging

3

7

Chandan Srivastava

3. Catalysis and Metal Sensing with Transiton Metal Complexes Immoblized on Nanopporous Micro-Scale and Nano-Scale Silica Polyamine Composites E. Rosenberg, Geoffrey Abbott and Abdul Goni

8

Invited Speakers 4. Potential Discrimination of Human Mesenchymal Stem Cells from Different Niches In-vitro and In-vivo by Raman Spectroscopy Opens New Horizons for Regenerative Medicine and Tissue Engineering T. Abruzzese, E. Brauchle, K. Schenke-Layland and W.K. Aicher 5. Potential Applications for Translational Nanotechnology Research and Education in the Pre-Hospital Poly-Trauma Environment Annette L. Sobel and Sharmilla Dissanaike 6. Tunning of the Surface Topography to Prevent Bacterial Adhesion Oscar Komla Awitor 7. Emerging and Re-Emerging Infectious Diseases: Nano/Micro-Scale Devices in Disease Control Basavaraj Madhusudhan 8. Multifunctional Nanoceramics and Their Composites: Applications in Energy Conversion, Memory Devices, Bioimaging and Drug Delivery Pankaj Poddar 9. Surface Modification of Magnesium Alloys for Biomedical Applications P. Amaravathy, A. Srinivasan and N. Rajendran 10. Using Stem Cells as a Model System to Investigate Toxicity of Silver Nanoparticles Pavan Rajanahalli

13

14 15

16

17 18 19

xx Contents

Contributed Papers 11. Biogenic Synthesis, Optimization, Characterization and Antibacterial Activity of Silver Nanoparticles Using Ocimum Sanctum Linn. Leaf Extract R. Ramachandran, C. Krishnaraj and P.T. Kalaichelvan 12. Structural and Optical Investigation of the Nanostructure on the Wings of Idea Malabarica (Moore, 1877) J. Sackey, Z.Y. Nuru, N. Delorme, B.T. Sone, S. Berthier and M. Maaza 13. Preparation of Zinc Oxide (ZnO) Nanoparticles Using Melia Dubia Leaf Extract and Its Charactraization S. Prabhu, K. Vaideki and S. Anitha 14. Tb3+ Doped Gadolinia Nanoparticles as Multifunctional Material for Biomedical Application Manisha Kumari, Rashmi Madhuri and Prashant K. Sharma 15. Biocompatibility Assessment of SiO2-TiO2 Composite Powder on MG63 Osteoblast Cell Lines for Orthopedic Applications M. Chellappa, B. Thejaswini and U. Vijayalakshmi 16. In-situ Fabrication of Nano-Composite and its Coatings by Electrophoretic Deposition on Ti-6Al-4V for Biomedical Applications M. Chellappa and U. Vijayalakshmi 17. Hemocompatibility Evaluation of Hydroxyapatite/Polymer based Scaffolds for Biomedical Applications S. Mary Stella, B. Priyadarshini, S. Arul Xavier Stango and U. Vijayalakshmi 18. Design and Development of Self Nanoemulsifying Drug Delivery System of Raloxifene Hydrochloride S.P. Sharavanan, P. Chandra Sekar, C. Senthil Kumar, S. Abimanyu, K. Venkateshwaran and N. Subramanian 19. Formulation Development and Evaluation of Solid Lipid Nanoparticles: As Ocular Drug Delivery System for Anti-Glaucoma Drugs C. Senthil Kumar, S.P. Sharavanan, P. Chandrasekar, P. Saravana Kumar and N. Subramanian 20. Resveratrol Loaded Chitosan/Pegylated Chitosan Nanoparticles: For Efficient Ocular Delivery P. Saravanakumar, P. Chandrasekar and N. Subramanian 21. Green Synthesis of Copper Oxide Nanostructures Using Morinda Citrifolia L. Fruit Extract: Optical and Electrical Studies S. Preethi, S. Vaishnavi, M. Sivakumar and N. Subramanian 22. Fabrication of a Novel 5% Ag Doped Hydroxyapatite-Fe3O4-Chitosan Hybrid Composites by Planetary Ball Milling Technique and Its In-vitro Biocompatibility Studies for Biomedical Applications U. Anjaneyulu, P. Manusha, J. Harathi and U. Vijayalakshmi 23. Formulation and In-vitro Evaluation of Stavudine Loaded Human Serum Albumin Nanoparticles J. Josephine Leno Jenita, K.S. Suparna, C. Vijaya, V. Murugan and Wilson Barnabas

23

27

33

37

41

47

53

57

61

65

69

73

79

Contents xxi

24. Cytotoxicity of Spherical Gold Nanoparticles Synthesized Using Aqueous Extract of Aerial Roots of Rhaphidophora aurea (Linden ex Andre) Intertwined over Areca catechu on MCF-7 Cell Lines 85 M. Jannathul Firdhouse and P. Lalitha 25. Green Synthesis of Metal Oxide Nanoparticles by Hibiscus Sabdariffa Extract as an Effective Chelating Agent 89 N. Thovhogi, A. Gurib-Fakim and M. Maaza 26. Synthesis and Characterization of Bioceramic/Triterpenoid Porous Nanocomposite for Biomedical Applications 95 J. Indira 27. Synthesis and Various Studies of Nano Powder of a Siddha Drug – Madhumega Choornam (MMC) 99 T. Gladys Vimala, V. Bena Jothy and P. Selvarajan 28. CNano Sensors for the Early Diagnosis of Cancer 105 D. Joslin Vijaya 29. Silver Nanoparticles: A Green Approach Mediated by Lantana Camara Leaf Extract 109 S. Hemanth Kumar, K. Hari Krishna, K. Roja Rani and P.J. Goutam 30. Preparation and Characterization of Atorvastatin and Curcumin Combination-Loaded Chitosan Nanoformulation for Oral Delivery in Atherosclerosis 113 J.B. Varuna Kumara, Sistla Ramakrishna and Basavaraj Madhusudhan 31. In-vitro Osteogenic Effect of Naringin Loaded Albumin Nanoparticles on C3H10T1/2 Mouse Mesenchymal Stem Cell Line 119 R. Ranjith, S. Balraj and M.C. John Milton 32. Development and In-vitro Evaluation of Oxytetracycline-loaded PMMA Nanoparticles for Oral Delivery against Anaplasmosis 123 T.S. Lakshmi Narayana, Prahlad C. Ghosh and Basavaraj Madhusudhan 33. In-vitro Antioxidant and Xanthine Oxidase Inhibitory Activities using Ethanolic Extract Caesalpinia coriaria pods 129 V. Yamini Priya-, D. Anandhi and E. Manikandan 34. Green Synthesis of Zinc Oxide Nano Particles (ZnONPs) from the Leaves of Eugenia Jambolana and its Toxicty Effects in Embryos of Zebrafish (Danio rerio) 135 Subhaschandrabose Jeyabharathi, Kalimuthu Kalishwaralal, Krishnan Sundar and Azhaguchamy Muthukumaran 35. Preparation of Chitosan/Different Organomodified Clay Polymer Nanocomposites: Studies on Morphological, Swelling, Thermal Stability and Anti-Bacterial Properties 141 B.H. Nanjunda Reddy, V. Venkata Lakshmi and K.R. Vishnu Mahesh 36. Amine Functionalized Copper (II) Oxide Nanoparticles Based Impedimetric Immunosensor for CA125 147 Ragini Raghav and Sudha Srivastava 37. Synthesis and Characterization of New Conducting Biopolymer: Chitosan Grafted Poly (Aniline-Co-m-Chloroaniline) 151 R. Anandarasu, J. Vivekanandan, A. Jeeva and P.S. Vijayanand

xxii Contents 38. Green Synthesis, Characterization of Copper Nanoparticles Using Syzygium aromaticum L. and Their Efficacy against Exserohilum turcicum (Pass.) Leonard and Suggs 155 V.B. Nargund, Chikkanna Swamy, Hulagappa and Pradeep Manyam 39. Biological Synthesis of Silver Nanoparticles Using Turbinaria Ornata and It’s In-vitro Antioxidative Activity 159 R.R. Remya, S.R. Radhika Rajasree, L. Aranganathan, T.Y. Suman and S. Gayathri 40. Investigation of Antimicrobial Activities and Mechanism of Silver Nanoparticles Synthesized from Cantharanthus Roseus Extract 165 S. Senthil Kumar and L.C. Nehru 41. Polymer-Lipid Hybrid Nanocapsules Improves Lutein Solubility, Stability and Biovailability in Mice 169 Arunkumar Ranganathan and Vallikannan Baskaran 42. Nano Scale Analysis of Siddhadrugs in Essential Drug List 173 M.S. Shree Devi, P. SathiyaRajeswaran, M. Kannan, R. Vasudevan and S.D. Muralidass 43. Glucose Monitoring System Using Nanopellet 177 C. Rajasekaran, K.B. Jayanthi and M. Nirmala 44. Phytofabrication of Gold and Palladium Nanoparticles Using the Leaves of Polygala Javana DC 183 R. Malathi and V. Ganesan 45. Chemical and Green Synthesized Gold Nanoparticles Cytotoxic Effect In-vitro: A Comparative Study 189 R. Saranya and E. David 46. Crystal Structure of Hemoglobin from Painted Turtle (Chrysemyspicta) 195 V. Maheshwaran, V. Thiruselvam, K. Ravichandran and M.N. Ponnuswamy 47. Crystal Structure of Hemoglobin from Hedgehog (Paraechinusmicropus) 199 V. Maheshwaran, V. Thiruselvam, P. Sugumar, K. Saraboji and M.N. Ponnuswamy 48. Crystal Structure Determination of Hemoglobin Fromgoldfish (Carassiusauratus) 203 M. Mohamed Abubakkar and M.N. Ponnuswamy 49. Structural and Functional Studies of an Iron Storage Protein: Ferritin—A Vehicle for Nano Drugs 209 V. Thiruselvam, T.S. Kumarevel and M.N. Ponnuswamy 50. In-vitro and Corrosion Study of Hydroxy Apatite Coatings by Pulse Laser Deposition on Ti6Al4V and 316L Stainless Steel 213 S. Gnanavel, S. Ponnusamya and C. Muthamizhchelvan 217 51. Interaction of Biocompatible Bi2S3 Nanorods with Human Serum Albumin Selvaraj Naveenraj, Ramalinga Viswanathan Mangalaraja, Thangaraj Pandiyarajan and Sambandam Anandan 52. A Versatile Synthetic Polymer from Chitosan Cross Links with Salicylaldehyde Derivative —Aniline Nanocomposites for Biomedical Applications 223 R. Ida Malaeselvi, C. Ramachandra Raja and J. Priscilla 53. A Critical Review on Bio Nano Encapsulation of Nutraceuticals 229 S. Thiruchenduran, K. Uma Maheswari, T.N.V.K.V. Prasad and N. Supraja

Contents xxiii

54. Phytosynthesis of Silver Nanoparticles from the Rind Extract of Garciniamangostana and its Synergistic Effect with Antibiotics against Human Pathogenic Bacteria 233 R. Nishanthi, S. Malathi and P. Palani 55. Chitosan-Alginate Scaffolds for Seeding Mesenchymal Stem Cells 237 S.G. Kumbhar and S.H. Pawar 56. Effect of Nanoparticle Ferric Pyrophosphate in Iron Deficiency, Its Impact on Plasma Proteins In-vivo 241 Y.S. Bindu, G. Mitra, M. Muralidharan, J. Pinto, D. Srivastava, A.V. Kurpad and A.K. Mandal 57. Mesoporous Silica Nanoparticle Loaded with Curcumin Reduces the Cell Survival of MCF-7 Cells 243 Lakshminarasimhan Harini, Bose Karthikeyan, Sweta Srivatsava, Srinag B.S., Cecil Ross, Gnanakumar G., Rajagopalan S. and Thandavarayan Kathiresan 58. Multiminerals Substituted Hydroxyapatite Coating on Surface Treated Surgical Grade Stainless Steel for Orthopedic Applications 249 D. Rajeswari, S. Ramya, L. Kavitha and D. Gopi 59. Gold Polymer Janus Nano-Bridge for the Combined Photothermal and Photodynamic Therapy 255 Vijayakumar Shanmugam 60. Synthesis and Characterization of Neomycin Functionalized Chitosan Stabilized Silver Nanoparticles and Study its Antimicrobial Activity 257 R.K. Preethika, R. Ramya, M. Ganesan, S. Nagaraj and K. Pandian 61. Antibacterial and Catalytic Properties of Silver Nanoparticles Loaded Zeolite: Green Method for Synthesis of Silver Nanoparticles Using Lemon Fruit Juice as Reducing Agent 263 J. Selvamuthumari, S. Meenakshi, M. Ganesan, S. Nagaraj and K. Pandian 62. In-Situ Functionalization of Aniline Oligomer onto Layered Graphene Sheet and Study of its Application on Electrochemical Detection of Ascorbic Acid in Food Samples 269 S. Devasena, S. Meenakshi, R. Sayeekannan and K. Pandian 63. Observations on FeCl3 Influence in Nano-Hydroxyapatite Property Modifications 275 A. Nishara Begum, S. Aravindan and C. Deepa 64. Synthesis and Characterization of Mwcnt/TiO2/Au Nanocomposite for Photocatalytic Activity 277 V. Karthika and A. Arumugam 65. Synthesis of PEG Modified Chitosan Nanocapsules Loaded with Thymoquinone 281 S. Vignesh Kumar, E. Hemananthan, S. Harish and P. Renuka Devi 66. Sodium Polyacrylate (SPA) Enhanced FPIA Based Detection of Pesticide Residue with PPB/PPT Level Detection Limit 287 Swathi Korrapati, Phani Kumar Pullela and Vijayalakshmi 67. ZnO Nanostructures and their Potential Applications 293 M.A. Shah 68. Poly (Vinyl Alcohol)/Poly (Acrylamide-Co-Diallyldimethyl Ammonium Chloride) PH Sensitive Semi-IPN Hydrogels for Ciprofloxacin Hydrochloride Drug Delivery 297 Siva Sankar Sana and Vijaya Kumar Naidu Boya 69. Hydroxyapatite Alumina Nanocomposites for Bone Implant Applications 301 M. Rajkumar, S. Vignesh Raj, A. Kanakaraj, N. Meenakshi Sundaram and V. Rajendran

xxiv Contents 70. Novel Biomimetic Three Dimensional Porous CMC-Gel Hydroxyapatite Nanocomposites as Major Load Bearing Synthetic Bone Graft Chandrani Sarkar and Subhadra Garai 71. Cadmium Toxicity on Hydroxyapatite Nanocrystals V. Ochigbo, V.O. Ajibola, E.B. Agbaji, A. Giwa, P. Mannivasakan and V. Rajendran Author Index

305 307 313

PLENARY SPEAKERS

Structural Aspects of Protease Control in Health and Disease Robert Huber Max-Planck-Institut fuer Biochemie, Emeritusgruppe Strukturforschung, D-82152 Martinsried, Germany Technische Universität München, TUM Emeritus of Excellence, D-85747 Garching, Germany Universität Duisburg-Essen, Zentrum für Medizinische Biotechnologie, D-45117 Essen, Germany Cardiff University, School of Biosciences, Cardiff CF10 3US, UK

ABSTRACT As a student in the early nineteen sixties, I had the privilege to attend winter seminars organized by my mentor, W. Hoppe, and by M. Perutz, which took place in a small guesthouse in the BavarianAustrian Alps. The entire community of a handful of protein crystallographers assembled in a room which served as living and dining room and as auditorium for the lectures. Today structural biologists organize large congresses with thousands of attendants and there exist many hundreds of laboratories specialized in this field. It appears to dominate biology and biochemistry very visibly if we count covers in scientific journals displaying macromolecular structures. Structural biology was successful, because it was recognized that understanding biological phenomena at the molecular and atomic level requires seeing those molecules. Structural biology revealed the structure of genes and their basic mechanism of regulation, the mechanism of enzymes’ function, the structural basis of immune diversity, the mechanisms of energy production in cells by photosynthesis and its conversion into energy-rich chemical compounds and organic material, the mechanism that makes muscle work, the architecture of viruses and multienzyme complexes, and many more. New methods had an essential impact on the development of structural biology. Methods seemed to become available in cadence with the growing complexity of the problems and newly discovered methods brought biological problems within reach for researchers, a co-evolutionary process of the development of methods and answerable problems. An important additional incentive for structural biology came from its potential application for drug design and development by the use of knowledge of drug receptors at the atomic level. The commercial interest in application spurred this direction of research enormously. My lecture will start out with a very brief review of the history of protein crystallography and continue with our studies since 1970 on proteolytic enzymes and their control. Proteolytic enzymes catalyze a very simple chemical reaction, the hydrolytic cleavage of a peptide bond. Nevertheless they constitute a most diverse and numerous lineage of proteins. The reason lies in their role as components of many regulatory physiological cascades in all organisms. To serve this purpose and to avoid unwanted destructive action, proteolytic activity must be strictly controlled.

V. Rajendran, P. Prabu and K.E. Geckeler (eds.) Bio-Nanomaterials for Biomedical Technology, pp. 3–4 (2015)

4

Application of Nanostructured Materials for Energy and Environmental Technology

Control is based on different mechanisms which I will discuss and illustrate with examples of systems and structures determined in my laboratory: (a) by specific inhibition with natural and synthetic inhibitors (b) by enzymatic specificity (c) by activation from inactive precursors accompanied or not by allosteric changes (d) by co-localization of enzyme and substrate (e) by cofactor binding accompanied or not by allosteric changes (f) by controlled access to the proteolytic site. The regulatory principles offer new opportunities of intervention for therapeutic purposes and use in crop science. I then will let you share my experience with the foundation and development of two biotech companies with different business models, but both based on basic academic research in structural biology. Proteros (www.Proteros.com) offers enabling technology services for Pharma- and Crop science companies imbedding all steps of the workflow molecular and structural biology can provide and commands and uses its platform for the generation of leads from identified targets to in vivo Proof of Concept (PoC). Suppremol (www.Suppremol.com) specializes in the development of novel immune-regulatory therapeutics for the treatment of autoimmune diseases on the basis of a recombinant, soluble, nonglycosylated version of the human Fcγ receptor IIB and of receptor binding antibodies. Suppremol was recently acquired by Baxter International Inc. (NYSE:BAX) offering an ideal setting for its therapeutic projects.

KEYNOTE SPEAKERS

Nanoparticles for Bio-Imaging Chandan Srivastava Department of Materials Engineering, Indian Institute of Science, Bangalore

ABSTRACT Magnetic resonance imaging (MRI) is a cellular imaging technique that is widely used as a diagnosis tool in medical science. In spite of rendering an excellent imaging spatial resolution, MRI technique suffers from limited probe sensitivity. Probe sensitivity of MRI can be enhanced by using materials that act as image contrast enhancing agents during the MRI process. This talk will illustrate the potential of the following materials as contrast agents for MRI: (a) graphene oxide-Fe3O4 nanoparticle composite (b) MnFe2O4-Fe3O4 core-shell nanoparticles and (c) CoFe2O4-ZnO core-shell nanoparticles. In the case of the work on graphene oxide-Fe3O4 nanoparticle composites, it was observed that the GO-Fe3O4 composite framework that contains graphene oxide with least extent of reduction of the carboxyl groups and largest spacing between the graphene oxide sheets provides the optimum structure for yielding a very high transverse proton relaxivity value. In case of work on MnFe2O4Fe3O4 core-shell nanoparticles, it was observed that the proton relaxivity value obtained in the dispersion of the core-shell nanoparticles was considerably greater than the proton relaxivity value obtained in the presence of single phase nanoparticles of the core and shell phases. This highest value of transverse relaxivity in the case core-shell nanoparticles was due to the largest magnetic inhomogeneity created by the core-shell nanoparticles in the water medium surrounding it. In the work on CoFe2O4-ZnO core-shell nanoparticles, two different core-shell geometries were investigated. In one case, the core-shell geometry was made up of individual nanoparticles containing CoFe2O4 as core and ZnO as shell and in the other case, agglomerates of CoFe2O4 nanoparticles were encapsulated within ZnO capsules. It was observed that a fluorescent CoFe2O4-ZnO core-shell nanoparticles with the unique geometry in which CoFe2O4 ferrite nanoparticles agglomerates were present within ZnO capsules yields very high value of transverse proton relaxivity when compared to the proton relaxivity value exhibited by the individual core-shell nanoparticles.


Cataly ysis and d Metall Sensin ng with h Transsiton M Metal Complex C xes Imm moblizeed on Nanopp N porous Micro-S M cale an nd Nanoo-Scalee Silica Poolyamin ne Com mpositess E. Rosenberg, Geofffrey Abbott and Abduul Goni Departmen nt of Chemistry and a Biochemistrry, University off Montana, Missooula, MT 59802,, USA E-mail: [email protected]

AB BSTRACT We haave been stud dying the covaalent binding of luminescennt and catalytiically active transition t metal compllexes to silica polyamine coomposites (SP PC) with a view w towards appplications in electron e transfe fer chemiistry, metal seensing and biifunctional caatalysis. In thee case of the luminescent Ru complexees, Ru(CO O) (H) (L2) (L’2)] [PF6], (L2 = trans-22PPh3, L’ = η2-4,4’-dicarbboxy-bipyridinne L2 = transs2Ph2PCH P 2CH2COO OH, L’2 = bipyyridine; L2 = Phh2PCHCHPPhh2) (L’ = η2-5--amino-1, 10-pphenanthroline; L2 = trans-2PPh3 (1) L’2 = η2-4-carboxaldeehyde-4’-methhyl-bipyridine)) binding to the compositte me, whose maagnitude is deependent on thhe surfacce results in siignificant incrreases in excitted state lifetim naturee of the surfacce and the sizee of the silicaa particle to which w they are bound (microo- versus nanooparticlles) [1, 2]. This T effect wiill increase thhe developmeent of these materials m as photo-promote p ed reduciing agents. When W metal ionns in aqueous in solution arre exposed to these surfacees quenching or o enhanncement of thee luminescence is observed that is dependdent on the typpe of metal ioon and sensitivve to the binding consttant of the meetal to the com mposite surfacee (2). In a sepparate study off the binding of o NN. the PN

1

2

Fig. 1:: Structures of Immobilized I Coomplexes on SP PC V. Rajeendran, P. Prabu and K.E. Gecckeler (eds.) Bio-Naanomaterials for Biomedical Technology, T pp. 8–9 (2015)

3

Catalysis and Metal Sensing with Transiton Metal Complexes Immoblized...

9

pincer complexes of ruthenium, (C6H3NCH2P(t-butyl)2)N(Et)2Ru(H)(CO)Cl (3), bound to the composite surface. We observed catalytic dehydrogenation of alcohols to give esters, a reaction that usually requires the addition of strong base. Here the polyamine surface provides the necessary basic environment, demonstrating the proposed bifunctional catalysis. Details and directions for the future will be presented. Mechanistic aspects of the catalysis will be presented and a comparison of the relative efficiency of the nano- and micro-composites for catalysis of the Kneovenagel reaction will be presented [2].

REFERENCES [1] Abbott, G. Geoffrey; Brooks, Robert; Rosenberg, Edward; Terwilliger, Michelle; Ross, J.B. Alexander and Ichire, Ogar O.L., “Surface Bound Ruthenium Diimine Organometallic Complexes: Luminescence and Lifetime Organometallics 33 (2014) 2467–2478. (dx.doi.org/10.1021/om401153x). [2] Abbott, Geoffrey; Brooks, Robert and Rosenberg, Edward, “Investigations on the Structure and Properties of Silica-Polyamine Composites on the Nano and Micro-scales,” Appl. Poly. Sci. (2015) DOI:10.1002/APP42271.

INVITED SPEAKERS

Potential Discrimination of Human Mesenchymal Stem Cells from Different Niches In-vitro and In-vivo by Raman Spectroscopy Opens New Horizons for Regenerative Medicine and Tissue Engineering T. Abruzzese, E. Brauchle1,2, K. Schenke-Layland1,2,3 and W.K. Aicher Department of Urology, University of Tübingen Hospital, Tübingen, FRG Department of Women’s Health, University of Tübingen Hospital, Tübingen, FRG 2 Department of Cell and Tissue Engineering, Fraunhofer IGB Stuttgart, FRG 3 Department of Medicine/Cardiology, UCLA, Los Angeles, CA, USA E-mail: [email protected]

1

ABSTRACT Mesenchymal stromal cells (MSCs) have been applied in countlesspre-clinical and clinicl studies to support wound healing, to prevent fibrosis after ischemia, ameliorate autoimmune disorders and others. In our current studies we aim to investigatere generation of smooth and striated muscle tissue by autologous MSCs. To this end we compared proliferation and differentiation capacities of human MSCs from three different sources. After expansion using GMP-compliant procedures, we monitored the cells using flow cytometry, who letranscriptomegene arrays, qRT-PCR, immune blotting, immunocy to chemistry, and Raman spectroscopy. Functional studies were performed in a large animal stemcell implantation model. We identified that human MSCs from different sources can be discriminated based on the irtelomerlenghts, transcript to me, proteome, expression of intracellular or membrane-anchored molecules, differentiation capacities or proliferation kinetics. In addition, on a single cell-based analysis, MSC can be distinguished from several more mature cells such as fibroblasts or from cells of other differentiation lineages, and most interestingly from MSC isolated from other sources. Raman spectroscopy allowed to localize MSCs injected in tissue samples. This technique will therefore be further explored for different clinical and diagnostic applications involving MSCs.


Potential Applications for Translational Nanotechnology Research and Education in the Pre-Hospital Poly-Trauma Environment Annette L. Sobel and Sharmilla Dissanaike1 Department of Medical Education and Department of Electrical Engineering, Texas Tech University, Texas Tech University Health Sciences Center, Lubbock, Texas, USA 1 Department of Surgery, Texas Tech University, Texas Tech University Health Sciences Center, Lubbock, Texas, USA

ABSTRACT Texas Tech University Health Sciences Center, located in West Texas, offers the only Level I trauma care within a range of 200 square miles across the Texas Panhandle, Eastern New Mexico and far Southwest Texas. The facility provides care to a catchment area of 500,000 for trauma and one million for burn patients. In addition, we provide the most sophisticated combination trauma and chemical burn management for the petroleum industry across the entire Permian Basin spanning Texas and Eastern New Mexico. This facility is also home to a research-intensive environment with longstanding collaborations with the U.S. Army’s Institute for Surgical Research and expanding research in biofilms and nanomaterials for accelerated wound healing and translational research. Faculty interests span across polytrauma and neurocognitive domains in both basic and applied research. This presentation will describe the emerging translational research, educational, and entrepreneurial environment at these institutions and the potential application of results to both civilian and military health care settings.


Tunning of the Surface Topography to Prevent Bacterial Adhesion Oscar Komla Awitor C-Biosenss (EA-4676), IUT - Département Mesures Physiques, Université d'Auvergne Campus des Cézeaux, 5 avenue Blaise Pascal, CS 30086, 63178 Aubière Cedex E-mail: [email protected]

ABSTRACT Bacterial infection of in-dwelling medical devices may result in serious peri-implant diseases and cannot always be avoided by sterilization prior to implantation or treated by traditional antibiotics due to the increasing prevalence of antimicrobial resistance and biofilm formation. Infection has been reported on various medical devices such as contact lenses, central venous catheters and needleless connectors, endotracheal tubes, intrauterine devices, mechanical heart valves, pacemakers, peritoneal dialysis catheters, prosthetic joints, tympanostomy tubes, urinary catheters, and voice prostheses. The number of complications such as infections, inflammatory reactions or inadequate are sometimes associated with the use of these devices. The number of patients in need of implants has grown rapidly in recent years. Systemic treatment may be used to attempt to reduce these complications. Sometimes, however, these new therapies are ineffective because of their low concentration or penetration in the zone of maximum tissue response and in a number of cases, the drug used may be toxic to the patient. In this context, the idea of targeting more precisely these drugs in the direct environment of the implants has always been a focus of development of these therapeutic strategies. In the context of medical implants, the material itself appears to be a particularly useful vector for applying these strategies for local delivery of molecules of interest. Nanoporous materials are excellent candidates for the design of the rapeutic implants because the topography of the surfaces at the nanometer scale facilitates the integration of tissue by optimizing the interfacial interaction between the biomaterial and the biological environment. The senanoporous architectures are of considerable interest for implantable medical devices such as orthopedic implants, vascular stents, cardiac implants and dental implants. They might have anti-bacterial functions or be used as drug delivery systems. Tunning of the surface topography of the implant or surface chemistry can prevent bacterial adhesion, but in any case, the performance of the implant in fulfilling its purpose as a function-replacing device must be preserved. This means that, even if bacteria are repelled, host cells are not to be harmed in order to guarantee optimal integration of the implant into the living. The nanoporous membranes like titanium dioxide nanotube layersare good candidates to improve tissue growth while simultaneously reducing infection without the use of pharmaceutical drugs and to drive molecules of interest at the right place and accelerate healing.


Emerging and Re-Emerging Infectious Diseases: Nano/Micro-Scale Devices in Disease Control Basavaraj Madhusudhan Research Centre for Nanoscience and Technology, Department of Biochemistry and Food Technology, Davangere University, Davangere E-mail: [email protected]

ABSTRACT A new political and economic paradigm is emerging with the turn of the century, which is affecting the prevention, control, and eradication of animal and zoonotic diseases. Despite remarkable advances in medical research and treatments during the 21st century, infectious diseases remain among the leading causes of death worldwide for three reasons: (1) emergence of new infectious diseases; (2) reemergence of old infectious diseases; and (3) persistence of intractable infectious diseases. New findings show similarities among infectious agents that span different taxa and kingdoms, and this trend is bringing together infectious disease specialists who earlier did not consider them to have common interests in disease prevention and management. Nanobiotechnologies are clinically applicable and possess the potential to be useful in laboratory diagnosis of infections in general and viral infections in particular. The dispersing of drug-carrying nanoparticles, referred to as nanosponge, into the body. Such sponges can target specific cells and areas that have been affected by disease. Using nanotechnology, engineering researchers at the global level including University of MissouriColumbia have developed small but powerful devices capable of enhancing the delivery of drugs to treat life-threatening illnesses. Fundamental view of how engineered nanoparticles interact with living cells. Development of innovative methods for diagnosis using micro- and nano-particles, investigating patho-mechanisms of possible particle in disease monitoring and control appears to be a challenging task. Nanotechnology is functional in the design of biochips as they enable the diagnosis at the molecule and single cell level and hence serve as a great advance in molecular diagnostics. The long-term objective of drug delivery systems is the ability to target selected cells and/or receptors within the body. Nanotechnology is critical in reaching these goals will be discussed.


Multifunctional Nanoceramics and Their Composites: Applications in Energy Conversion, Memory Devices, Bioimaging and Drug Delivery Pankaj Poddar Physical & Materials Chemistry Division, National Chemical Laboratory, Pune

ABSTRACT The areas of ceramics and their composites have been industrially highly important with rich optical, magnetic and dielectric properties. Moreover, the ceramics are usually chemically stable and few of them are biocompatible too. We have studied the ceramics of various kinds at nanoscale for the magnetic, ferroelectric and optical properties. In iron oxide nanoparticles, we showed for the first time that the Verwey transition exist for the particles as small as 8 nm [1, 2]. We reported biosynthesis of BaTiO3 with size 5μg) was used as a source of zinc and was cleaned by ultra-sonication in acetone and water for 10 minutes in each solvent. A closed cylindrical Teflon lined stainless steel chamber was used for the synthesis. 4mg of zinc metal foils was taken with 40 ml of de-ionized water in a Teflon-lined stainless steel chamber with 50 ml capacity. The prepared reaction mixture was kept at 120°C in an oven for 15 hours. After the desired time, the system was allowed to cool down naturally and the resulting mixture was centrifuged. The zinc foils, collected from the reactions vessels, were washed with de-ionized water several times and finally dried in air. Characterization Phase structure and the purity of the as prepared samples were characterized by powder X-ray diffraction (XRD) taken on a Philips (X’Pert PRO PW-3710) diffractometer with 2θ ranging from 10– 70°, using Cu Kα (λ = 0.15141 nm) radiation operated at 40kV and 30mA. The morphology of the products was carried out using Field Emission Scanning Electron Microscope (FEI SEM, NNL 200, Japan), coupled with energy dispersive X-ray spectrometer EDX (Gensis). RESULTS AND DISCUSSION Structural Information The XRD patterns of the prepared samples synthesized at 120°C shown in Figure 1 The diffraction peaks are obtained at (100), (002), (101), (102), (110) and (112) planes which are characteristic of the pure ZnO with the wurtzite hexagonal phase. All the peaks in the pattern have been indexed to hexagonal wurtzite structure with space group P63mc and lattice constants a = 0.3249 nm, c = 0.5206 nm, (JCPDS card no. 36–1451). No diffraction peaks arising from any impurity has been detected in the pattern confirms that the grown products are pure ZnO. The relative broad peaks suggest high crystallinity of the samples

Fig. 1: XRD Pattern of As-Grown Sample

ZnO Nanostructures and their Potential Applications

295

Morphological Examinations The general morphologies of the as-grown structures, obtained after the reaction of zinc metal with water at 100°C for 15 h, was observed by FESEM and demonstrated in figure 2 which confirms that the grown products are hexagonal nanorods in shape. Figure 2 (a) and (b) show the low-magnification FESEM images of the nanorods and confirms that the nanorods are grown in a very high density over the whole foil substrate.

Fig. 2: Typical (a) Low and (b) High-Resolution FESEM Images of Nanostructures

The Formation Mechanism The formed ZnO nuclei are the building blocks for the formation of the final products. Even though a plausible growth process for the formation of ZnO hexagonal-shaped ZnO nanorods are described here but more studies are needed to clearly explain the growth process for the formation of these nanorods. Due to crystal habits of ZnO, the nuclei have a hexagonal shape. In the wurtzite hexagonal phase, ZnO has polar and nonpolar faces. In polar ZnO crystals, the zinc and oxygen atoms are arranged alternately along the c-axis and the top surfaces are zinc terminated (0001) and are catalytically active while the bottom surfaces are O-terminated (0001) and are chemically inert. The zinc metal on reaction with water slowly gives out hydrogen (g) and the liberated oxygen reacts with metal to give oxides as shown in the above reaction. The Zn reacts with oxygen and forms ZnO nuclei, which further serve as seeds for ZnO nanorods growth.

Fig. 3: Schematic Illustration of the Growth (a) Zn Particles and Water Vapour Co-exists. (b) Layer of Zn(OH)2 Formed on the Particle Surface and Subsequent Decomposition to ZnO

CONCLUSION In summary, hexagonal ZnO nanorods were prepared by water in a simple reaction of Zn powder and water at 90°C without any organics. The performance of the as synthesized rods was investigated and their excellent bacterial effect is demonstrated. These can be recommended for the purification of medical and food equipments. This method is found to be a mild, convenient and efficient route to prepare ZnO nanostructures without the template or crystal seeds. It may be extended to fabricate other metal oxide nanomaterials.

296

Bio-Nanomaterials for Biomedical Technology

ACKNOWLEDGEMENT The Author (Shah M.A) is pleased to acknowledge the financial support of World Bank for procurement of Scanning Electron Microscope. REFERENCES [1] [2] [3] [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] [14] [15]

Gao, P.X. and Wang, Z.L., J Am Chem Soc 2003; 125: 11299–305. Shen, G.Z., Bando, Y. and Lee, C.J., J Phys Chem B 2005; 109: 10779–85. Park, W.I., Yi, G.C., Kim, M. and Pennycook, S.J., Adv Mater 2002; 14: 1841–3. Vayssieres, L., Keis, K. and Hagfeldt, A., Lindquist SE. Chem Mater 2001; 13: 4395–8. Vayssieres, L., Adv Mater 2003; 15: 464–6. Lao, J.Y., Huang, J.Y., Wang, D.Z. and Ren, Z.F., Nano Lett 2003; 3: 235–8. Kong, X.Y. and Wang, Z.L., Nano Lett 2003; 3: 1625–31. Peng, W.Q., Qu, S.C., Cong, G.W. and Wang, Z.G., Cryst Growth Des 2006; 6: 1518–22. Tian, Z.R., Voigt, J.A., Liu, J., Mckenzie, B. and Mcdermott, M.J., J Am Chem Soc 2002; 124: 12954–5. Wei, A., Sun, X.W., Xu, C.X., Dong, Z.L., Yu, M.B. and Huang, W., Appl Phys Lett 2006; 88: 213102. Gao, X.P., Zheng, Z.F., Zhu, H.Y., Pan, G.L., Bao, J.L., Wu, F., et al. Chem Commun 2004; 12: 1428–9. Cao, J.M., Wang, J., Wang, B.Q., Chang, X., Zheng, M.B. and Wang, H.Y., Chem Lett 2004; 33: 1332–3. Verianshyah, B., Park, T.J., Lim, J.S. and Lee, Y.W., J Solid Supercritical fluids, 34 (2005) 51–61. Shah, M.A., Al-shahri, M. and Assiri, A.M., Int. j. of Nanopart. (IJNP), Vol. 2, No. 2 66 (2009). Al-Harbi, L.M. and Shah, M.A, Growth of ZnO nanorods and their optical properties, Mater. Appl. Sci. 5, No. 2, 87–91 (2011).

Poly (Vinyl Alcohol)/Poly (Acrylamide-CoDiallyldimethyl Ammonium Chloride) PH Sensitive Semi-IPN Hydrogels for Ciprofloxacin Hydrochloride Drug Delivery Siva Sankar Sana and Vijaya Kumar Naidu Boya Department of Materials Science and Nanotechnology, Yogi Vemana Univeristy, Kadapa, Andhra Pradesh E-mail: [email protected]

ABSTRACT Herein, we developed pH responsive polyvinyl alcohol (PVA), acryl amide (Am) and Diallyldimethyl ammonium chloride (DADMAC) semi-interpenetrating polymer network (semiIPN) hydrogels by radical polymerizationusing N, N’-methylene-bis-acrylamide (MBA) as a crosslinker and ammonium persulphate (APS) as an initiator. The resulting hydrogels were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and X-ray diffraction (XRD) studies. In vitro release of ciprofloxacin hydrochloride (CFH) from hydrogel was performed in acidic and basic media.

INTRODUCTION Hydrogels are three-dimensional cross-linked hydrophilic polymer chains, which swells when contact with water or other biological fluids [1]. Now-a-days, most of the research in this area is focused on the design of multi polymer pH sensitive semi-IPN hydrogel systems because of their better performance than that of individual polymers and are sensntive to external conditions [2]. The pHsensitivity of the matrix is mainly coming from the presence of functional groups such as weakly acidic (-COOH) and basic (-NH2) the polymer backbone[3]. Among the different polymers studied, PVA stood in a prime position because of its non carcinogenicity, good biocompatibility and a high degree of swelling in aqueous solutions. Hence in this paper, we developed multi functional pH sensitive polyvinyl alcohol (PVA), acryl amide (Am) and diallyldimethyl ammonium chloride (DADMAC) semi-IPN and loaded the ciprofloxacin hydrochloride (CFH). CFHdrug affects bacterial DNA gyrase and topoisomerase IV, without affecting mammalian DNA activity [5]. EXPERIMENTAL METHODS Semi-IPNhydrogels were prepared by a free radical polymerization and the composition of hydrogelsarepresented in Table 1. Required amounts of monomers, Am and DADMAC were dissolved in 3 mL of distilled water and then required amount of PVA solution, MBA and APS were added. The solution was maintained at 55°C for 3h to complete the reaction under nitrogen gas. The obtained gels were kept in distilled water for about a week by frequently changing the water to remove the unreacted monomers and APS. Then, the gels were dried in vacuum oven at 40°C.0.1 M solution V. Rajendran, P. Prabu and K.E. Geckeler (eds.) Bio-Nanomaterials for Biomedical Technology, pp. 297–300 (2015)

298


of CFH was loaded into the hydrogel by immersing the hydrogel in drug solution for 24 h and dried at ambient temperature. Table 1: Formulation Code and Composition of Semi-IPN Gels PVA (2%) mL

Am (gm)

MBA (mg)

DADMAC (mL)

SIPN1

2

0.5

30

0.25

33.26

0.606

2.5292

SIPN2

2

0.5

30

0.5

35.45

0.599

2.9376

SIPN3

2

0.5

30

1.0

37.68

0.589

3.4197

SIPN4

2

0.5

15

0.5

40.75

0.595

3.3265

SIPN5

2

0.5

50

0.5

30.43

0.608

SIPN6

1

0.5

30

0.5

34.85

0.606

2.5585

SIPN7

3

0.5

30

0.5

38.25

0.590

3.3419

Code

% EE

n

k

2.3067

CHARACTERIZATION OF SEMI-IPN HYDROGELS FTIR spectra of hydrogels was recorded on Perkin Elmer Spectrophotometer whereas DSC on a SHIMADZU DSC-60. XRD pattern of the gels were recordedon X-ray diffractometer(RIGAKU, SMARTLAB) operated at 30kV 100mA with Cu Kα as a radiation source.SEMimages were taken using HITCHI S-3700N. SWELLING EXPERIMENTS The equilibrium selling studies were carried out at pH 1.2 and 7.4 buffersolutions and equilibrium swelling ratio (ESR)was determined by using the equation (1), % ESR

W

W W

100

… (1)

where, Wd is the weight of the drygel and We is theWeight of the gel after establishment of equilibrium

ENCAPULATION EFFICIENCY AND IN VITRO DRUG RELEASE STUDIES About 10 mg of the drug-loaded hydro gels were added in 10mL of buffer solution and stirred vigorously for 2 days to extract drug from the hydro gels. The solution was filtered and assayed by UV spectrophotometer (Lab India, Mumbai, India) at fixed max value of 275 nm. The results of % drug loading (DL) and encapsulation efficiency (EE) were calculated, using equations (3) and (4) respectively and included in Table 1. In vitro drug release studieswere carried out on Tablet dissolution Tester (LAB INDIA, Mumbai, India). Dissolution rates were measured at 37± 0.5 oC at constant speed of 100 rpm for 12 hrsin pH1.2 and 7.4 phosphate buffer solutions. DL %

W W

100 (2)EE %

A T

100(3)

RESULTS AND DISCUSSION FTIR spectra of semi-IPN gels was shown in Figure 1. A reveals the peaks at 3437, 2923, 2854 cm– 1 which corresponds to O-H and C-H stretching vibrations, where as peaks at 1122 cm−1, 1260 and1610 cm–1can be attributed to C–N,C–F andphenyl group which is conjugated to –COOH,

Poly (Vinyl Alcohol)/Poly (Acrylamide-Co-Diallyldimethyl Ammonium Chloride) PH Sensitive...

299

respectively.Peak appeared at 3421 cm–1was attributed to N–Hvibrations.These peaks also appeared in drug loaded gel. Pure CFH drugs shows a sharp melting peak at 155 oCwhereas no peak was observed in the drug loaded gel at this temperaturein DSC thermogramsshown in Figure 1B. XRD data reveals the peaks at 11.48, 19.43, 26.68 and 29.46 odue to the crystalline nature of CFH drug in drug but no such peaks are not found in CFH loaded hydrogels, indicating that the drug is dispersed at molecular level in the hydrogel networks presented in Figure 1.C..The SEM image showsthat porous nature in itsstructure.

Fig. 1: A. FTIR Spectrum, 1.B. DSC Curves, 1.C. XRD Spectrum and SEM Image of Gels

Fig. 2(A): Equilibrium Swelling of Semi-IPNgels at pH 1.2 and 7.4 and (B) % Cumulative Release of CFH at 7.4 pH Medium

300


As the amount of MBA increases the swelling ratio and % cumulative release of CFH decreases in the hydrogels which is due to increase in rigidity of the gel.As the amount of DADMAC and PVA increases the swelling ratio and % cumulative release of drug increasesdue to presence of hydrophilic moieties on monomers which in turn increases the overall hydrophilic nature of the gel. The equilibrium swelling ratio of semi- IPN gel is more at pH 7.4 due to physical adsorption of hydroxyl groupscompared to pH 1.2 [6]. Figure 2 shows that effect of MBA, DADMAC and PVA on the CFH % cumulative release profile at pH 7.4 and 37 oC.The release kinetics of CFH from cumulative release data (Mt/M∞) with respect to time by fitting the data to equation (4), Mt/M∞ = ktn(4). Here, Mt/M∞ represents the fractional drug release at time t, n is a diffusion parameter characterizing the release mechanism, and k is a constant characteristic of the drug-polymer system. Using the least squares procedure, the k and n values were estimated for the different formulation. The results are included in Table 1.

CONCLUSION In this paper, PVA/Poly (Am-co-DADMAC) semi-IPN hydro gels were prepared and characterized by FTIR, DSC, XRD and SEM. Drug release studies indicated that, the developed hydrogels showed controlled release nature in releasing CFH drug and also showed that the charactrstics of hydrogles can be altered by changing the external physical conditions. ACKNOWLEDGEMENT Authors (BVKN & SSS) are thankful to Department of Science and Technology, New Delhi for their financial support (SR/FT/CS-70/2010). REFERENCES [1] Wenbo, W. and Aiqin, W., Synthesis and swelling properties of pH-sensitive semi-IPN super absorbent hydrogels based on sodium alginate-g-poly (sodium acrylate and polyvinylpyrrolidone, Carb. Polym, 80, 1028–1036, 2010. [2] Hu, X., Feng, L., Wei, W., Xie, A., Wang, S., Zhang, J. and Don, W., Synthesis and Characterization of a novel semi-IPN hydro gel based on Salecan and poly (N, N-dimethylacrylamide-co-2hydroxyethylmethacrylate, Carb. Polym., 105, 135–144. 2014., [3] Ravindra, S., Antoine, F., Mulaba-Bafubiandi, Rajineekanth, V., Varaprasad, K. and Mohana Raju, K., Synthesis of Surfactant-Modified Poly (Acrylamide-co- Potassium Acryl ate) Hydro gels and Its in vitro Release Studies, Polym.-Plast. Technol. Eng., 51, 1355–1360, 2012. [4] Singh, B. and Pal, L. J. Sterculiacrosslinked PVA and PVA-poly (AAm) Hydrogen wound dressings for slow drug delivery: Mechanical, Mucoadhesive, biocompatible and permeability properties the mechanic. Biomed. Materials, 9, 9– 21, 2012. [5] Marchese, A.L., Slana, V.S., Holmes, E.W. and Jay, W.M., Toxicity and Pharmacokinetics of Ciprofloxacin, J. Ocul. Pharmacol., 9, 69–76. 1993. [6] Chen, J., Liu, M., Liu, H., Ma, L., Gao, C., Zhu, S. and Zhang, S., Synthesis and properties of thermo- and pH-sensitivepoly(diallyldimethylammoniumchloride)/poly(N, N-diethylacrylamide) semi-IPN hydrogel Chem, Eng., 159, 247–256, 2010.

Hydroxyapatite Alumina Nanocomposites for Bone Implant Applications M. Rajkumar, S. Vignesh Raj1, A. Kanakaraj, N. Meenakshi Sundaram2 and V. Rajendran3 Department of Physics, PSG College of Arts and Science, Coimbatore, Tamil Nadu Department of Biomedical Engineering, PSG College of Technology, Coimbatore, Tamil Nadu 2 Department of Physics, Government Arts College, Salem, Tamil Nadu 3 Centre for Nanoscience and Technology, K.S. Rangasamy College of Technology, Tiruchengode, Tamil Nadu E-mail: [email protected] 1

ABSTRACT Recently, the combination of biomimetic and bioactive materials plays a vital role in the field of biomedical engineering. Especially, hydroxyapatite (HAp) and its hybrid composites have significant impact on bone replacement and coatings on implant surfaces. In this work, the bioinert alumina (Al2O3) was incorporated with the biomimetic bone mineral hydroxyapatite for the preparation of hydroxyapatite-alumina nanocomposites under hydrothermal conditions. The structural and morphological analysis of the prepared composites was discussed using X-ray diffractrometer and electron microscopical studies. Further, the thermal stability and biocompatibility studies were performed to study the influence of alumina into the hydroxyapatite bioceramic. These ceramic-ceramic composite prepared under low temperature hydrothermal condition proves to be an effective implant material for bone replacement. Keywords: Hydroxyapatite, Alumina, Hydrothermal, Biocompatibility, Bone Implants.

INTRODUCTION In the recent years, biomimetic hydroxyapatite (HAp) [Ca10(PO4)6(OH)2]and its composites received significant attention in bone implantation and replacements [1]. Reinforcement of metal oxides into this HAp to form nanocomposites provides an efficient route for bone regeneration and hard tissue engineering applications. Addition of metal oxides such as alumina, ziroconia and titania favors the formation of apatite structures [2]. Several methods were employed to synthesis nanocomposites of HAp-metal oxide composites such as protein foaming-consolidation method [3], underwater shock compaction [4] and sol-gel [5]. In this work, we demonstrated the synthesis of HAp-alumina nanocomposites using hydrothermal method. Further, the cytotoxicity study has been made on prepared nanocomposites against human osteosarcoma cell line MG-63 which has similar osteogenic property like primary osteoblasts cells. EPERIMENTAL PROCEDURE Preparation of Hydroxyapatite-Alumina (HAp-Al2O3) Composite The HAp-Al2O3 composites was hydrothermally prepared using calcium nitrate tetrahydrate (Ca(NO3)2.4H2O, purity 98%, Merck, Mumbai), diammonium hydrogen phosphate (NH4)2HPO4, V. Rajendran, P. Prabu and K.E. Geckeler (eds.) Bio-Nanomaterials for Biomedical Technology, pp. 301–304 (2015)

302


purity 99%, Merck, Mumbai) and aluminium nitrate nonahydrate (Al2(NO3)2.9H2O, >95%, Merck, Mumbai) as starting materials. All the reagents were analytical grade and used without further treatment process. The stoichiometric ratio of Ca/P was maintained at 1.67 and 0.5 M of aluminium nitratenonahydrate) was used for the preparation of HAp-Al2O3 composite. The pH of the reaction solution was adjusted to 11 using 25% ammonia solution. The HAp and alumina precursors were added into the hydrothermal vessel and treated at 250°C for 3h under autogenous pressure. The hydrothermally prepared powder was dried and calcined at 900 °C for 3 h in a muffle furnace. The calcined composite powders were stored in an air tight container and used for further analysis.

RESULTS AND DISCUSSION The X-ray Diffraction (Shimadzu, XRD-6000, Japan) patternof HAp and HAp-aluminacomposite are shown in Figure 1(a) and Figure 1(b) respectively. In Figure 1(a), the predominant peaks of HAp around 32° was observed without any additional peaks for impurities whereas the XRD pattern of HAp-Al2O3 composite as shown in Figure 1(b) reveals the broadening of characteristics peaks of HAp. This may be due to the influence of alumina addition in to the HAp matrix. This led to the assumption that the reinforcement of alumina into the HAp matrix favors the formation of HAp-Al2O3 composite. However, no characteristic peaks for alumina was observed as some phases of alumina exhibits amorphous nature [5].

Fig. 1: XRD Pattern of (a) Pure HAp Calcined at 700 °C for 3h (b) HAp-Alumina Composite Calcined at 900 °C for 3h

Figure 2(a) and 2(b) shows the Fourier Transform Infrared (FTIR, Perkin Elmer, USA) spectra of pure HAp and HAp/alumina composite respectively. The bands at 565, 600 and 958 cm–1 corresponds to PO43– and the strong band at 1056 cm–1 assigned to the P–O stretching vibration of PO43–. The stretching vibration and bending modes of the OH–appear at 3571 cm–1 and 633 cm–1 corresponds to typical stoichiometric HAp.

Fig. 2: IR Spectrum of (a)Pure HAp and (b) HAp-Alumina Composite

Hydroxyapatite Alumina Nanocomposites for Bone Implant Applications

303

The band at 3439 cm–1 may come from lattice H2O. The characteristic bands for inorganic carbonate ion (1460 and 875 cm–1) are present in the FTIR spectra of pure HAp which appeared due to the atmosphere carbon absorption as shown in Figure 2(a) [5].

(a)

(b)

(c)

(i) Pure HAp

(d)

(e)

(f)

(ii) HAp-alumina Composites Fig. 3: HRTEM and EDS Results of (i) Pure HAp and (ii) HAp-Alumina Composite

The High resolution transmission electron microscope (HRTEM, JEOL-JEM, 2100, Japan) images and EDS results of pure HAp and HAp-alumina composite are shown in Figure 3(i) and Figure 3(ii) respectively. Figure 3(a) and 3(b) shows the nanostructures of pure HAp with small agglomerates. In Figure 3(d) and 3(e) of HAp-Alumina composites micrographs, nanospheres and nanorods were observed which may corresponds to HAp and alumina respectively hence one dimensional nanostructures of metal oxides can be easily formed by hydrothermal process without addition of surfactants/stabilizing agents. In addition, the EDS results confirm the presence of characteristic elements in the prepared samples as shown in Figure 3(c) and 3(f). The cytotoxicity of prepared HAp-Alumina nanocomposite was performed using MTT assay against human osteosarcoma cell line MG-63. The results show 80% cell viability for the maximum concentration (200µg/ml) as shown in Figure 4. The MTT assay confirms the compatibility of the prepared HAp-Alumina composite material for bone implant applications.

304


Fig. 4: Cytotoxicity Evaluation of Prepared HAp-Alumina Composites against MG-63 Human Osteosarcoma Cell Line Using MTT Assay

CONCLUSION In the present work, pure HAp and HAp-Alumina composite were successfully prepared under low temperature hydrothermal process at relatively minimum time. The micrographs of the prepared HApAlumina composite shows agglomerated nanorods and nanospheres which reveals the non uniform morphological distribution. In addition, reinforcement of alumina does not affect the biocompatibility of HAp which was examined by 80% cell viability against human osteosarcoma cell line MG-63. Based on these results, we suggest that HAp-Alumina composites prepared under low temperature hydrothermal process might suitable for bone restoration applications. ACKNOWLEDGEMENTS The author acknowledges the University Grants Commission (UGC), New Delhi for the financial support and PSG institutions for providing the necessary facilities to carry out the research project. REFERENCES [1] Hannora, A.E., Preparation and Characterization of Hydroxyaptite/Alumina Nanocomposites by HighEnergy Vibratory Ball Milling. Journal of Ceramic Science and Technology, 5(4), 293–297, 2014. [2] Mezahi, F.Z., Oudadesse, H., Harabi, A. and Gal, Y., Effect of ZrO2, TiO2, and Al2O3 additions on process and kinetics of bonelike apatite formation on sintered natural hydroxyapatite surfaces. International Journal of Applied Ceramic Technology, 9(3), 529–540, 2012. [3] Sopyan, I., Fadli, A. and Mel, M., Porous alumina–hydroxyapatite composites through protein foaming– consolidation method. Journal of the mechanical behavior of biomedical materials, 8, 86–98, 2012. [4] Chiba, A., Kimura, S., Raghukandan, K. and Morizono, Y., Effect of alumina addition on hydroxyapatite biocomposites fabricated by underwater-shock compaction. Materials Science and Engineering: A, 350 (1), 179–183, 2003. [5] Yelten, A., Yilmaz, S. and Oktar, F.N., Sol–gel derived alumina–hydroxyapatite–tricalcium phosphate porous composite powders. Ceramics International, 38(4), 2659–2665, 2012. [6] Labat, B., Chamson, A. and Frey, J., Effects of γ‐alumina and hydroxyapatite coatings on the growth and metabolism of human osteoblasts. Journal of biomedical materials research, 29(11), 1397–1401, 1995.

Novel Biomim N B metic Th hree Diimensioonal Poorous C CMC-G Gel Hyd droxyapatite Nanoco N omposittes as Majorr Load Bearin ng Syntthetic Bone B Grraft Channdrani Sarkkar and Subbhadra Garrai CSIR— —National Metaallurgicallaborattory, Jamshedpuur E-maill: chandrani@nm mlindia.org; [email protected]

INTR RODUCTION N To mimic m the nattural architecture of bonee a wide rannge research on polymer hydroxyapatitte compoosites is being g continued, yet the mechanical propertty of polymerr hydroxyapattite compositees have not n been matcched with natuural bone [1].. Thus metals and alloys arre used as im mplants showinng stress shielding, infflammation annd complexityy after implanntation. Howeever, nanocom mposites shoulld m strrength with reegenerative prroperty whichh makes it suittable for majoor possesss sufficient mechanical load bearing b orthop pedic application. Inn this work, we w focused onn the synthessis of novel thhree dimensioonal polymer--hydroxyapatitte nanoccomposites th hrough a veryy simple andd cost effectiive route witthout adding any chemical crosslinker. PRAC CTICAL SIN NGNIFICAN NCE Biodeegradable poly ymer Carboxyymethyl Celluulose (CMC) and Gelatin (Gel) ( a derivaate of collageen have been used, fo or in-situ form mation of maajor load bearring CMC-Geel-HA nanocoomposites. Thhe bondinng between two t polymerss create a strrong chemicaally bound orrganic matrixx where in-sittu minerralization of HA H occurred [2]. The miicrostructures obtained froom SEM exhhibited that thhe polym mer concentrattion increasess, crosslinkingg increases. As A a result thhe agglomeration increasestto produuce compact microstructuree of nanocom mposites. Thee nanocompoosites achieveed compressivve strenggth and elastiic modulus in i the range of 8–86 MP Pa and 0.10––1.2 GPa resspectively. Thhe crosslinking betweeen polymerscoonfirmed from m FTIR spectrra. Itplays the main role forr enhancing thhe BF mechaanical compreessive strengthh of nanocompposites. Biodeegradability off nanocompossites under SB (simullated body flluid) solutionn indicated coontinuous redduction in meechanical streength and alsso exhibiited bone apattite depositionn.

Fig. 1: 1 SEM Image of o Synthesized Nanocomposite N es V. Rajeendran, P. Prabu and K.E. Gecckeler (eds.) Bio-Naanomaterials for Biomedical Technology, T pp. 305–306 (20155)

306


Fig. 2: Stress/Strain Curve of Synthesized Nanocomposites

Fig. 3: FTIR Spectra of Synthesized Nanocomposites

Fig. 4: Bone Apatite Deposition and Compressive Strength Reduction (in inset) of Nanocomposite after SBF Immersion

CONCLUSION We have synthesized novel biomimetic three dimensional porous CMC-Gel-Hydroxyapatite nanocomposites having high compressive strength and elastic modulus by simple and cost effective method. Synthesized nanocomposites exhibited bone regenerative property and have great potential as synthetic bone graft for major load bearing application. REFERENCES [1] Dhandayuthapani, B. et al., International Journal of Polymer Science 2011, Article ID 290602, 19 pages, 2011. [2] Garai, S. and Sinha, A., Biomimeticnanocomposite of Carboxymethylcellulose-hydroxyapatite: Novel three dimensional load bearing bone grafts, J Colloids and Surface B:Biointerfaces, 115, 182–190, 2014.

Cadmium Toxicity on Hydroxyapatite Nanocrystals V. Ochigbo, V.O. Ajibola1, E.B. Agbaji1, A. Giwa2, P. Mannivasakan3 and V. Rajendran3 National Research Institute for Chemical Technology (NARICT), Zaria-Nigeria 1 Department of Chemistry, Ahmadu Bello University Zaria-Nigeria 2 Department of Textile Science and Technology, Ahmadu Bello University Zaria-Nigeria 3 Centre for Nanoscience and Technology, K.S. Rangasamy College of Technology, Tiruchengode, Tamil Nadu, India E-mail: [email protected]

ABSTRACT Nanocrystalline bioactive hydroxyapatite ceramic powder was synthesized using the wet precipitation (WT) and hydrothermal (HT) techniques. The toxicity of Cadmium ion (Cd2+) on the hydroxyapatite was also studied. Calculated amounts of Cadmium nitrate and chloride solutions were used as the dopants on the hydroxyapatite powder. The resultant powders obtained were analysed using different characterization techniques. The results indicated that irrespective of the techniques employed in the synthesis of the hydroxyapatite and the source of Cadmium ion (Cd2+) dopants, they both gave crystalline powders and Cadmium ion (Cd2+) exchanged well with the Calcium ion (Ca2+) of the hydroxyaptite via adsorption and dissolution ion exchange mechanism, but the crystallinity decreases in the order: hydrothermal (HT) > wet precipitation (WT) techniques and Cadmium nitrate (CdN) > Cadmium chloride (CdC) sources. Keywords:Cadmium, Toxicity, Hydroxyapatite, Nanocrystals, Wet precipitation, Hydrothermal, Techniques, Nitrate, Chloride, Dopants.

INTRODUCTION Hydroxyapatite (HA) [Ca10(PO4)6(OH)2] is a naturally occurring mineral in the inorganic component of human bone and tooth enamel in a nano range. The constituent elements of HA are primarily calcium and phosphorus, with hydroxide ions that are eliminated at elevated temperatures and a stoichiometric Ca/P ratio of 1.667. Hydroxyapatite (HA) is the most researched calcium phosphate biomaterial. This biomaterial is widely used to repair, fill, extend and reconstruct damaged bone tissue [1, 2]. The apatite (HA) structure is so tolerant to ionic substitutions that Ca2+ in the crystals can be replaced by various divalent cations including Mg2+, Cd2+, Sr2+, Pb2+ and Ba2+[3–6]. Cadmium ion (Cd2+) shows a negative effect on bone tissues. However, its mechanism is not fully understood. One of the possible reactions involves interaction between cadmium and calcium, which results in calciuria and leads to a reduction in calcium absorption from intestines. Nevertheless, it has been reported that a relationship exists between the increased content of cadmium in the body and a decreased level of mineral density of the bones, increased bone resorption and decreased levels of the parathyroid hormone. This metal influences trace elements that affect bone tissue, mainly due to the interaction among zinc, iron, and copper [7]. Cadmium accumulated in bones can affect the activity of V. Rajendran, P. Prabu and K.E. Geckeler (eds.) Bio-Nanomaterials for Biomedical Technology, pp. 307–312 (2015)

308


bone cells and directly increase the loss of mineral elements in the bones, as it is known to activate osteoclasts and inhibits osteoblasts [8]. The toxic activity of cadmium is not only limited to bone tissue quantity reduction only but also to its quality deterioration. Cadmium exposure also results in a decreased level of types I and V collagen in the bones and leads to their increased solubility by damaging the intermolecular cross-links [9]. A change in the collagen structure decreases its endurance, thus increasing susceptibility to deformation and fractures. This study is aimed at investigating the toxicity of Cadmium ion (Cd2+) from two sources of cadmium salts at series of concentrationson the structural properties of hydroxyapatite synthesized via wet precipitation (WT) and hydrothermal (HT) techniques at physiological conditions.

MATERIALS AND METHODS Materials Analar grade Calcium Nitrate tetra hydrate (Ca(NO3)2.4H2O), Diammonium hydrogen phosphate ((NH4)2HPO4) and liquid ammonia (NH3) and Nitric Acid (HNO3) from Merck chemical were used. Methods Pure and doped nanocrystalline HA powders were synthesized through the wet precipitation and hydrothermal techniques according to the method reported [10] but with some modifications. In wet precipitation synthesis, the aging was done at room temperature for 24 hours, while in the hydrothermal aging was done in an autoclave for 24 hours at 150°C. The suspension was then allowed to cool naturally after which the white precipitate obtained on aging both samples were then centrifuged, filtered under mild suction, washed with distilled deionised water and dried in an oven at 80oC for 24 hrs. The HA crystals obtained with these methods were ground with a mortar and pestle and calcined at 300ºC in a muffle furnace under an air atmosphere for 3 hr. In order to synthesize nanocrystalline HA powder doped with Cadmium, 0.5g of HA was added to a 25 ml aliquot of 0.2, 0.4 and 0.6M Cadmium nitrate and chloride solution respectively followed by over night continuous shaking of suspension at 37oC in an electrical incubator shaker. After achieving saturation ion exchange equilibrium, the suspension was centrifuged, filtered under mild suction with little washing and dried using air oven at 80°C overnight. The pure and doped HA were then characterized. Characterizations The particle size distribution was determined with a sub micrometer particle size analyzer (Nanophox, Sympatec, Clausthal-Zellerfeld, Germany) according to the dynamic light scattering techniques. Elemental compositions of the prepared HA and doped HA samples were ascertained using X-ray fluorescence (XRF) spectrometer (EDX-720; Shimadzu, Kyoto, Japan). The presence of functional groups was confirmed through Fourier transform infrared (FTIR) spectrometer in the range of 4000– 400 cm−1 using KBr as a reference.The structural nature of HA and doped HA powders were characterized through X-ray diffraction (XRD) analysis using an X-ray diffractometer (X’ PertPro, PANalytical, Almelo, the Netherlands) withCuKα as the radiation (λ = 1.5418 Å) source. The source was operated at 40 kV with 2Ө value varying from 10o to 80o and the average crystallite size was estimated from the Debye- Scherrer approximation.

Cadmium Toxicity on Hydroxyapatite Nanocrystals

RESULTS AND DISCUSSION

HT HA

WT HA

(C) HT CdC

(D) WT CdC

(E) HT CdN

(F) WT CdN

Fig. 3: FT-IR Spectra of HA Synthesized Via (A) Hydrothermal Techniques (B) Wet Precipitation Techniques and Doped with Cadmium Chloride (CdC) and Cadmium Nitrate (CdN) Respectively at Physiological Conditions

309

310


Counts

Counts

HT0_5B

W T0_5B

150

100

100

50 50

0

0 20

30

40

50

60

70

20

80

30

40

50

60

Position [°2Theta] (Copper (Cu))


(A) HT

(B) WT

Counts

70

80

C o unts W TCdC

HT CdC 100 10 0

50

50

0

0 20

30

40

50

60

70

20

80

30

40

50

60

70

80

P o sition [°2T he ta ] (C o pp er (C u))

P osition [°2T heta] (Copper (Cu))

(C) HT CdC

(D) WT CdC Counts

Counts

HTCdn

WTCdn

150

150

100

100

50

50

0

0 20

30

40

50


(E) HT CdN

60

70

80

20

30

40

50

60

70

80


(F) WT CdN

Fig. 4: XRD Pattern of HA Synthesized Via (A) Hydrothermal Techniques (B) Wet Precipitation Techniques and Doped with Cadmium Chloride (CdC) and Cadmium Nitrate (CdN) Respectively at Physiological Conditions

Discussion The projected techniques for the synthesis of nano hydroxyapatite (HA) through hydrothermal and wet precipitation methods as well HA doped with Cadmium from Chloride and Nitrate sources were compared for their average particle size, phase purity, crystallinity and crystal size. Between these methods, hydrothermal method is more suitable for synthesis of nano hydroxyapatite under

Cadmium Toxicity on Hydroxyapatite Nanocrystals

311

experimental conditions as it gave the apatite with the least average particle size of 26 nm compared to the 29 nm obtained via wet precipitation method. The results of semi quantitative chemical analysis acquired by X-ray fluorescence and energydispersive X-ray spectroscopy (XRF/EDS) reveals stoichiometric HA (Ca/P = 1.73) for hydrothermal and (Ca/P = 1.65) for wet techniques respectively close to the theoretical value (Ca/P = 1.67). This discrepancy in the (Ca/P) ratio could be attributed to the elevated aging temperature of hydrothermal compared to the wet precipitation method. The presence of Cd2+ was also detected in all the samples except for the pure HA as the concentration of Cadmium solution increases, there is associated decrease in the calcium ion concentration of the HA while the Ca/P ratio increases and (Ca+Cd)/P ratio is nearly a constant irrespective of the source of the cadmium ion. The functional groups of the compound were identified using Fourier transforms informed spectroscopy (FT-IR). Here its evident that doped HA shows significant changes from the pure HA with notable changes seen as a decrease in the hydroxyl stretching bands at 3572 cm–1 and 631 cm–1 and the phosphate stretching band at 962 cm–1 on changing the cadmium source from chloride to nitrates and the synthetic route from hydrothermal to wet precipitation respectively. The formation of hydroxyapatite nano particle was confirmed by X-ray diffraction (XRD). It is observed that the amount of cadmium ion adsorbed onto the HA synthesized via hydrothermal is less when compared to that adsorbed onto HA via wet precipitation method. This could be attributed to the nature of reaction of HA in the metal solution. It is thus inferred that the HA obtained through hydrothermal method in reaction with the metal ion solution follows the adsorption of ions on the apatite surface, followed by subsequent release of the cation originally contained within the solid mechanism while the HA obtained via wet precipitation method follows the dissolution of the apatite in the aqueous solution containing the donor ions followed by precipitation or co-precipitation mechanism. It also revealed that the cadmium ion (Cd2+) from the nitrate source is adsorbed more onto the apatite than those from the chloride source irrespective of the synthetic route for the apatite preparation.

CONCLUSION Toxicologically, HA prepared via hydrothermal method is preferred to that obtained via wet precipitation since it adsorbed less of the dopants from the metal solution which invariably means it will adsorb less amount of such metal from the total burden of it present in the bone when used as artificial bone or as a bone filler or implants. Also, since the cadmium ion is adsorbed more from the nitrate source onto the HA, it then implies more toxicity than from the chloride source counterpart. This discrepancy in the adsorption rate could be attributed to the reactivity of these anions based on the electronegativity and position on the reactivity series. REFERENCES [1] Yfantis, C.D., Yfantis, D.K., Depountis, S., Anastassopoulou, J. and Theophanides, T., Academic Environment of Biomaterials Science and Engineering at the School of Chemical Engineering of NTUA, 5th WSEAS/IASME International Conference on Engineering Education (EE’08), Greece, ISSN: 1790– 2769, 2008 pp. 27–32. [2] Salemi, H., Behnamghader, A., Afshar, A., Ardeshir, C.M. and Forati, T., Calcium Phosphate Formation on Alkaline-Treated Titanium by Biomimetic Synthesis, Proceedings of the 2007 WSEAS Int. Conference on Cellular and Molecular Biology - Biophysics & Bioengineering, Greece, 2007, pp. 126–129. [3] Elliott, J.C., Structure and Chemistry of the Apatites and Other Calcium Orthophosphates Elsevier, Amsterdam, 1994, p. 111.

312


[4] Brown, P.W. and Constantz, B., Hydroxyapatite and Related Materials, CRC Press, Inc., Boca Raton, 1994, p. 3. [5] Cheng, Z.H., Yasukawa, A., Kandori, K. and Ishikawa, T., J. Chem. Soc. Faraday Trans., 94. (1998), 1501. [6] Yasukawa, A., Ouchi, S., Kandori, K. and Ishikawa, T., J.Mater. Chem., 6 (1996), 1401. [7] Wilson, A.K. and Bhattacharyya, M.H. (1997). Effects of cadmium on bone: an in vivo model for the early response. Toxicol Appl Pharm, 145:68–73. [8] Akesson, A., Bjellerup, P., Lundh, T. et al. (2006). Cadmium-induced effects on bone in a populationbased study of woman. Environ Health Persp 114:830–834. [9] Galicka, A., Brzóska, M.M., Średzińska, K. et al. (2004). Effect of cadmium on Collagen content and solubility in rat bone. Acta Biochim Pol 51:825–829. [10] Syed SibteAsghar ABIDI and Qasim MURTAZA (2013). Synthesis and Characterization of Nano hydroxyapatite powder using wet chemical precipitation. U.P.B Sci. Bull., Series B, Vol. 75, Iss. 3.

Author Index 8

Abbott, Geoffrey Abimanyu, S. Abruzzese, T. Abubakkar, M. Mohamed Agbaji, E.B. Aicher, W.K. Ajibola, V.O. Amaravathy, P. Anandan, Sambandam Anandarasu, R. Anandhi, D. Anitha, S. Anjaneyulu, U. Aranganathan, L. Aravindan, S. Arumugam, A. Awitor, Oscar Komla

57 13 203 307 13 307 18 217 151 129 33 73 159 275 277 15

Balraj, S. Barnabas, Wilson Baskaran, Vallikannan Begum, A. Nishara Berthier, S. Bindu, Y.S. Boya, Vijaya Kumar Naidu Brauchle, E.

119 79 169 275 27 241 297 13

Chandrasekar, P. Chellappa, M. David, E. Deepa, C. Delorme, N. Devasena, S.

61, 65 41, 47 189 275 27 269

Devi, M.S. Shree Devi, P. Renuka Dissanaike, Sharmilla Firdhouse, M. Jannathul Ganesan, M. Ganesan, V. Garai, Subhadra Gayathri, S. Ghosh, Prahlad C. Giwa, A. Gnanakumar, G. Gnanavel, S. Goni, Abdul Gopi, D. Goutam, P.J. Gurib-Fakim, A. Harathi, J. Harini, Lakshminarasimhan Harish, S. Hemananthan, E. Huber, Robert Hulagappa Indira, J. Jayanthi, K.B. Jeeva, A. Jenita, J. Josephine Leno Jeyabharathi, Subhaschandrabose Jothy, V. Bena Kalaichelvan, P.T.

173 281 14 85 257, 263 183 305 159 123 307 243 213

8 249 109 89 73 243 281 281 3 155 95 177 151 79 135 99 23

314

Author Index

Kalishwaralal, Kalimuthu Kanakaraj, A. Kannan, M. Karthika, V. Karthikeyan, Bose Kathiresan, Thandavarayan Kavitha, L. Korrapati, Swathi Krishna, K. Hari Krishnaraj, C. Kumar, C. Senthil Kumar, P. Saravana Kumar, S. Hemanth Kumar, S. Senthil Kumar, S. Vignesh Kumara, J.B. Varuna Kumarevel, T.S. Kumari, Manisha Kumbhar, S.G. Kurpad, A.V. Lakshmi, V. Venkata Lalitha, P.

135 301 173 277 243 243 249 287 109 23 57, 61 61 109 165 281 113 209 37 237 241 141 85

Maaza, M. 27, 89 Madhuri, Rashmi 37 Madhusudhan, Basavaraj 16, 113, 123 Mahesh, K.R. Vishnu 141 Maheshwaran, V. 195, 199 Maheswari, K. Uma 229 Malaeselvi, R. Ida 223 Malathi, R. 183 Malathi, S. 233 Mandal, A.K. 241 Mangalaraja, Ramalinga Viswanathan 217 Manikandan, E. 129 Mannivasakan, P. 307 Manusha, P. 73 Manyam, Pradeep 155

Meenakshi, S. Milton, M.C. John Mitra, G. Muralidass, S.D. Muralidharan, M. Murugan, V. Muthamizhchelvan, C. Muthukumaran, Azhaguchamy

263, 269 119 241 173 241 79 213 135

Nagaraj, S. Narayana, T.S. Lakshmi Nargund, V.B. Naveenraj, Selvaraj Nehru, L.C. Nirmala, M. Nishanthi, R. Nuru, Z.Y.

257, 263 123 155 217 165 177 233 27

Ochigbo, V. Palani, P. Pandian, K. Pandiyarajan, Thangaraj Pawar, S.H. Pinto, J. Poddar, Pankaj Ponnusamya, S. Ponnuswamy, M.N. Prabhu, S. Prasad, T.N.V.K.V. Preethi, S. Preethika, R.K. Priscilla, J. Priya, V. Yamini Priyadarshini, B. Pullela, Phani Kumar Raghav, Ragini Raj, S. Vignesh

307 233 257, 263, 269 217 237 241 17 213 195, 199, 203, 209 33 229 69 257 223 129 53 287 147 301

Author Index

Raja, C. Ramachandra Rajagopalan, S. Rajanahalli, Pavan Rajasekaran, C. Rajasree, S.R. Radhika Rajendran, N. Rajendran, V. Rajeswaran, P. Sathiya Rajeswari, D. Rajkumar, M. Ramachandran, R. Ramakrishna, Sistla Ramya, R. Ramya, S. Ranganathan, Arunkumar Rani, K. Roja Ranjith, R. Ravichandran, K. Reddy, B.H. Nanjunda Remya, R.R. Rosenberg, E. Ross, Cecil Sackey, J. Sana, Siva Sankar Saraboji, K. Saranya, R. Saravanakumar, P. Sarkar, Chandrani Sayeekannan, R. Schenke-Layland, K. Sekar, P. Chandra Selvamuthumari, J. Selvarajan, P. Shah, M.A. Shanmugam, Vijayakumar Sharavanan, S.P.

223 243 19 177 159 18 301, 307 173 249 301 23 113 257 249 169 109 119 195 141 159

315

Sharma, Prashant K. Sivakumar, M. Sobel, Annette L. Sone, B.T. Srinag, B.S. Srinivasan, A. Srivastava, Chandan Srivastava, D. Srivastava, Sudha Srivatsava, Sweta Stango, S. Arul Xavier Stella, S. Mary Subramanian, N. Sugumar, P. Suman, T.Y. Sundar, Krishnan Sundaram, N. Meenakshi Suparna, K.S. Supraja, N. Swamy, Chikkanna

37 69 14 27 243 18 241 147 243 53 53 57, 61, 65, 69 199 159 135 301 79 229 155

Thejaswini, B. Thiruchenduran, S. Thiruselvam, V. Thovhogi, N.

41 229 195, 199, 209 89

Vaideki, K. Vaishnavi, S. Vasudevan, R. Venkateshwaran, K. Vijaya, C. Vijaya, D. Joslin Vijayalakshmi Vijayalakshmi, U. Vijayanand, P.S. Vimala, T. Gladys Vivekanandan, J.

33 69 173 57 79 105 287 41, 47, 53, 73 151 99 151

7

8 243 27 297 199 189 65 305 269 13 57 263 99 293 255 57, 61