CHAPTER 1 INTRODUCTION

1.1 Biomaterials:
Biomaterial can be defined as "any substances (other than a drug) or combination of substances synthetic or natural in origin, which can be used for any period of time, as a whole or as a part of a system which treats, augments, or replaces any tissue, organ or function of the body.[Von Recum et. al, 1995] Commonly, any matter, surface, or construct that interacts with biological systems is termed as biomaterial. Theoretically, any material can be a biomaterial as long as it serves the stated medical and surgical purposes. Biomaterials can be synthetic or natural materials that are required or designed to function appropriately in biological environment. Materials that constitute parts of medical implants, extracorporeal devices, and disposables that have been utilized in medicine, surgery, dentistry, and veterinary medicine as well as in every aspect of patient health care.

1.2 Classification of biomaterials: 1.2.1Metallic biomaterials:

Metallic implants provide the necessary strength and toughness required in load-bearing parts of the body. Due to these advantages, metals will continue to play an important role as orthopedic biomaterials in the future, even though there are concerns with regard to the release of certain ions from and corrosion products of metallic implants. Titanium (Ti) is one of the best biocompatible metals and is used most widely as an implant. Recent studies revealed an even greater biomedical potential for porous metals. Besides Ti, there are other metallic implants made of pure Zr, Hf, V, Nb, Ta, Re,Ni, Fe, Cu, Ag, stainless steels and various alloys suitable for biomedical application (Sergey V. Dorozhkin et al,2011)

Metallic materials continue to play an essential role as biomaterials to assist with the repair or replacement of bone tissue that has become diseased or damaged (Puleo DA et al ,1995) Metals are more suitable for load-bearing applications compared with ceramics or polymeric materials due to their combination of high mechanical strength and fracture toughness. Currently approved and commonly used metallic biomaterials include stainless steels, titanium and cobalt chromium-based alloys. A limitation of these current metallic biomaterials is the possible release of toxic metallic ions and/or particles through corrosion or wear processes that lead to inflammatory cascades which reduce biocompatibility and cause tissue loss. Moreover, the elastic moduli of current metallic biomaterials are not well matched with that of natural bone tissue, resulting in stress shielding effects that can lead to reduced stimulation of new bone growth and remodeling which decreases implant stability (Nagel j. l,2003 ) Table 1: Summary of the physical and mechanical properties of various implant materials in comparison to natural bone Properties Density(g/cm3 ) Elastic modulus(Gpa) Compressive Y.S(Mpa) Fracture Toughness(Mpam1/2) Natural Bone 1.8-2.1 3-20 130-180 3-6 Mg 1.74-2.0 41-45 65-100 15-40 Ti 4.4-4.5 110-117 758-1117 55-115 Co-Cr 8.3-9.2 230 450-1000 NA SS 7.9-8.1 189-205 170-310 50-200 Synthetic HA 3.1 73-117 600 0.7

. Fig 1.1 Example of metallic implants: dental implants and knee joint implant

1.2.1.1Stainless steels
Stainless steel used for medical implants is mainly austenitic type 316L due to its corrosion resistance along with a wide range of other physical and mechanical properties coupled with inert surfaces. The chemical composition of type 316L austenitic stainless steel where L denotes low carbon content is as follows: 0.030 % C, 1.0 % Si, 2.0 % Mn, 0.045 % P, 0.030 % S, 12.015.0 % Ni, 16.0-18.0 % Cr, and 2.0-3.0 % Mn (Ratner et. al, 2004]. The chemical composition of type 316L stainless steel was developed to obtain stable austenitic structure which has numerous advantages, namely: 1. Austenitic stainless steel has a face centered cubic structure and is characterized by very low yield strength to tensile strength ratio and high formability 2. To increase strength, cold working and successive strain aging treatment can be applied. 3. Austenitic stainless steel is superior to ferritic stainless steel in corrosion resistance because the crystallographic atomic density of the former is higher than that of the latter. 4. Austenitic stainless steel is nonmagnetic.

Drawbacks
The disadvantages of austenitic stainless steels generally are higher sensitivity toward pitting corrosion and stress corrosion cracking [Sumita et. al, 2004]. Pitting is accelerated by the existence of an oxygen concentration cell at the early growth stage. The corrosion resistance is improved by adding molybdenum, increasing nickel and reducing carbon to less than 0.030 %.This steel has less than 0.03 wt.% carbon in order to reduce the possibility of in vivo corrosion. If the carbon content of the steel significantly exceeds 0.03 %, there is a tendency for formation of carbides such as Cr23C6 which precipitate at grain boundaries. This carbide precipitation results in depletion of chromium from the grain boundary regions, which has the effect of diminishing the formation of the protective chromium-based oxide Cr2O3 [Bombac et. al, (2007]. Due to high content of chromium, 316L stainless steel forms a protective, adherent and coherent oxide film that envelops the entire outer surface. The film passive in nature serve as a barrier to corrosion processes in alloy systems that would otherwise experience very high corrosion rates [Liu C et. al, 2003] and has ability of self-healing, when damaged, as chromium in the steel reacts with oxygen and moisture in the environment to reform the protective oxide layer [Newson et. al, 2002].

1.2.1.2 Cobalt alloys

Cobalt alloys are nonmagnetic, corrosion and heat resistant and exhibit high strength even at elevated temperatures and are also resistant to wear [Davids et. al, 1998]. The superior properties are attributed to the crystallographic nature of cobalt, and formation of extremely hard carbides and the corrosion resistance imparted by chromium. Due to their excellent resistance to degradation in the oral environment, the first medical use of cobalt alloys was in the cast of dental implants. Various in-vitro and in-vivo tests have shown that the alloys are biocompatible and suitable for use as surgical implants. Today the use of Co alloys for surgical applications is mainly related to orthopaedic prostheses for the knee, shoulder and hip as well as to fracture fixation devices. Nowadays the CoCrMo alloys are known to be highly biocompatible materials [Long, M. et. al, 1998] and are widely used as orthopaedic implant materials for hip joints and knee replacement. The biocompatibility of CoCrMo alloy is closely related to its excellent corrosion resistance due to
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the presence of an extremely thin passive oxide film that spontaneously forms on the alloy surface. Similar to AISI 316L stainless steel predominant oxide film is Cr2O3 with some minor contribution from Co and Mo oxides [Kocijan et. al, 2004].

Drawbacks
Despite their excellent corrosion resistance, there is still some concern about metal ion release from orthopedic implants into the human body environment. Implant components fabricated from Co-Cr based alloys have been reported to produce elevated Co, Cr and Ni concentrations in the surrounding tissue [Okazaki Y et. al (2005]. The use of Cobalt alloys has been limited due to the difficulty in fabrication of these alloys. Other Co alloys used in medicine are CoNiCrMo (ASTM F 562) with a nickel content of 35 % used for cardiovascular pacing leads, stylets, catheters and orthopedic cables. Increased content of nickel exhibits an improved resistance to stress-corrosion cracking in aqueous solution, but also increase the possibility of nickel allergy reactions. Therefore these alloys are not ideal for orthopedic applications. The biocompatibility of the wear particles produced can be troublesome because of the increased surface area of these small particles which are in direct contact with the surrounding medium or tissue material. In work-hardened or work-hardened and aged conditions, this alloy has very high tensile properties which are among the strongest available for implant applications. Other Co-based alloy is L-605 cobalt alloy or CoCrWNi (ASTM F 90) which is used for heart valves and in an annealed condition (ASTM F 1091) for surgical fixation wires. Its mechanical properties are approximately the same as those of Co-CrMo alloys, but after the material is cold worked the mechanical properties more than double.

1.2.1.3 Mg and Its alloys:

The increase in the purity of Mg can affect the Mg structure and corrosion in the following ways:

(a) Fewer impurities results in fewer intermetallics, leading to the reduction of galvanic corrosion. Mg is anodic compared to the impurities; therefore the impurities are actually protected by sacrificing Mg. (b) Mg is an active metal which forms an oxide layer on the surface. Though this oxide layer is generally believed to have limited effect on the corrosion of Mg, an increased purity will reduce the number of defects in the oxide layer that might help to suppress pitting corrosion. They found that it is possible to modify corrosion kinetics by microstructural changes in the bulk or in the near surface region without any further processing( B. Denkena, et al,2007). Another tool to tailor the corrosion rate of pure Mg grains is their crystallographic orientation. Lu et al. showed experimentally that grains near (0 0 0 1) orientation are the most corrosion resistant Inoue et al., found that high purity Mg (99.9999% by weight) showed a lower corrosion rate than low purity Mg (99.9%) in a conventional chloride solution. They also concluded that in buffered chloride solution the corrosion rate does not depend on the purity of the material or on the alloying composition, but solely on the pH of the solutions (C. Opt Hoog et al,2008) Currently, controlling Mg properties for implant applications such as strength and corrosion resistance is achieved by alloying the pure metal with a variety of different elements [13,20]. This practice brings some material challenges which are summarized as the following: (1) A small amount of alloying element added to pure Mg metal increases the corrosion rate, which is well documented (X. Gu et all,2009); (2) Polycrystalline Mg and related alloys are routinely machined from cast ingots. They reveal grain boundaries, surface roughness, and structural defects, which are more vulnerable to corrosion . Lower corrosion rate is preferred because it does not affect the cell viability . The grains tend to segregate impurities along their boundaries thus amplifying the corrosion rate. The end result is enhanced evolution of hydrogen that causes local gas cavities in vivo (3) The majority of the alloying elements employed to improve the mechanical properties of Mg are toxic at high local concentrations if released from corrosion sites: Ni, Be, 110 Cu, Li, Al, Zn, Mn, Biodegradable Mg coating prepared by Physical Vapor Deposition (PVD) technique with
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controlled grain size and morphology is the one which helps us to control the corrosion rate in mg biomaterials. (M. Liu et al, 2008)

1.2.2 Ceramic Biomaterials:

Metal oxide ceramics, such as alumina (Al2O3, high purity, polycrystalline, fine grained), zirconia (ZrO2) and some other oxides (e.g., TiO2, SiO2) have been widely studied due to their bioinertness, excellent tribological properties, high wear resistance, fracture toughness and strength as well as relatively low friction. (Vallet-Regi l ,2006) Polymeric biomaterials face the problem of uncontrolled degradation, material release and issues with durability. Ceramic materials have been developed as alternatives that remain mostly insoluble and can be removed only by specific cell activity that is by steoclast cells. Besides they are ideal for use as substrates for bone mineralization by osteoblast cells. Zirconia (ZrO2) is bio-inert nonresorbable oxide with good chemical and dimensional stability, and a high strength and toughness. Zirconia ceramic material is potentially suitable for the highly loaded environments found in joint replacement. Zirconia ceramic has been used to manufacture femoral heads for total hip replacements since the late 1980s. Its colour and excellent biocompatibility and mechanical properties make it an attractive choice for dental applications. A prerequisite for successful bone implant integration is direct bone apposition which was observed at bone-zircona interfaces in histological and ultrastructural studies suggesting that ZrO2 may also be a suitable implant material. Biomedical grade zirconia exhibits the best mechanical properties of oxide ceramics but is also prone to ageing in the presence of water. Alumina ceramic (Al2O3) is one of the most widely used materials for implants and prostheses. It belongs to the first class of biomaterials with excellent bio inertness. The material is characterized by its excellent biocompatibility and high strength, hardness and fracture resistance. Its excellent wear resistance makes it widely used in femoral joints. However, alumina due to its inertness fails to cause any bone growth in the surroundings causing subsequent loosening and hence is not suitable for implant components with bone contact.

Bioactive glasses have also been used in many medical applications. However, due to their poor mechanical properties, these glasses cannot be used in load-bearing applications, where metallic alloys are still the materials of choice. One of the main applications of bioactive is as coatings on prosthetic metallic implants. The coatings serve the following two purposes: 1. Improve the osseointegration ability of the implants, 2. Act as barrier between metal and body tissues, thus protecting body from the corrosion products of the implants and protect metal from corrosion by the body fluids. However their use is limited due to their poor adhesion properties as coatings on the metallic implants. Other materials like transition metal nitrides, like TiN, ZrN, TiAlN, NbN, TaN and VN have been proven effective as coatings against wear and corrosion of surgical implants and prosthesis.

1.2.3 Polymeric biomaterials:

Their properties can be fine-tuned over a wide range by varying the type of polymer or chain length as well as by copolymerization or blending of two or more polymers. Unlike ceramics, polymers exhibit substantial viscoelastic properties and easily can be fabricated into complex structures, such as sponge-like sheets, gels or complex structures with intricate porous networks and channels. Being X-ray transparent and non-magnetic, polymeric materials are fully compatible with modern diagnostic methods, such as CT and magnetic resonance imaging. Unfortunately, most of them are unable to meet the strict demands of the in vivo physiological environment. Namely, the main requirements for polymers suitable for biomedical applications are that they must be biocompatible, not elicit an excessive or chronic inflammatory response upon implantation and, for those that degrade, they must breakdown into non-toxic products only. Unfortunately, polymers, for the most part, lack of rigidity, ductility and, ultimately, the mechanical properties required in load-bearing applications. Thus, despite their good biocompatibility, many of the polymeric materials are mainly used for soft tissue replacements (such as skin, blood vessel, cartilage, ligament replacement, etc.). Moreover, the sterilization processes (autoclave, ethylene oxide and 60 Co irradiation) may affect the polymer properties. (Langer R.et al, 2000)
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There are a variety of biocompatible polymers suitable for biomedical applications.123,124 For example, polyacrylates, poly(acrylonitrile-co-vinylchloride) and polylysine have been investigated for cell encapsulation and immunoisolation.(chen .H et al.2008)

1.2.4. Composite material

Composite materials are engineered materials made from two or more constituent materials with significantly different physical or chemical properties, which remain separate and distinct on a macroscopic level within the finished structure (wang m et al, 2003) Thus, composites are always heterogeneous. Furthermore, the phases of any composite retain their identities and properties and are bonded, which is why an interface is maintained between them. This provides improved specific or synergistic characteristics that cannot be obtained by any of the original phases alone. (Gibson et al, 2010) Hydroxyapatite is chemically similar to the mineral component of bones and hard tissues in mammals. It is one of few materials that are classifieded as bioactive, means that it would support bone ingrowth and osseointegration when used in orthopaedic, dental and maxillofacial applications.The chemical nature of hydroxyapatite lends itself to substitution, meaning that it is not uncommon for non-stoichiometric hydroxyapatites to exist. The most common substitutions involve carbonate, fluoride and chloride substitutions for hydroxyl groups, while defects can also exist resulting in deficient hydroxyapatites (Victor F et al,2004) The ideal bone graft should be benign, available in a variety of forms and sizes, all with sufficient mechanical properties for use in load-bearing sites, form a chemical bond at the bone/implant interface as well as be osteogenic,osteoinductive, osteoconductive, biocompatible, completely biodegradable,at the expense of bone growth, and moldable to fill and restore bone defects. Further, it should resemble the chemical composition of bones (thus, the presence of calcium orthophosphates is mandatory), exhibit contiguous porosity to encourage invasion by the live host tissue as well as possess both viscoelastic and semi-brittle behavior, as bones do. Moreover, the degradation kinetics of the ideal implant should be adjusted to the healing rate of the human tissue, with absence of any chemical or biological irritation and/or toxicity caused by substances that are released due to corrosion or degradation. (Doblare et al,2004)
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1.3 Magnesium as biomaterial:

Magnesium is an exceptionally lightweight metal. With a density of 1.74 g/cm3, magnesium is 1.6 and 4.5 times less dense than aluminum and steel, respectively]. The fracture toughness of magnesium is greater than ceramic biomaterials such as hydroxyapatite, while the elastic modulus and compressive yield strength of magnesium are closer to those of natural bone than is the case for other commonly used metallic implants (Okuma et al 2001). Moreover, magnesium is essential to human metabolism and is naturally found in bone tissue . It is the fourth most abundant cation in the human body, with an estimated 1 mol of magnesium stored in the body of a normal 70 kg adult, with approximately half of the total physiological magnesium stored in bone tissue. Magnesium is a co-factor for many enzymes, and stabilizes the structures of DNA and RNA . The level of magnesium in the extracellular fluid ranges between 0.7 and 1.05 mmol/L, where homeostasis is maintained by the kidneys and intestine. Enhance the material's strength and fatigue resistance owing to grain refinement (Vormann et al , 2003)

1.4 Ball milling:

Top down approach of material processing Suitable for alloying process without thermal energy Nano size particle can be produced known technique to achieve uniform mixing of powders Appropriate amount of milling times with varying milling speeds have been deployed to achieve various functions: particle size reduction, uniform mixing of powders, micro alloying, and composite formation and also phase changes in case of high energy milling. Ball milling involves cold welding, fracturing and rewelding of powder particles.

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Fig 1.2 Mechanism of ball milling Mechanical milling has been used to produce oxide-dispersion strengthened (ODS) nickeland iron-base superalloys for applications in the aerospace industry.

It has now been shown to be capable of synthesizing a variety of equilibrium and nonequilibrium ahoy phases starting from blended elemental or prealloyed powders. Suryanaryana et. al, 2003

1.5 Biological testing:

To test the performance of materials in biological environments, several invitro and invivo tests are done. The ability of material to bond with the bones is tested using Simulated body fluid by checking the hydroxyapatite formation on the surface. The ion concentration in SBFs is equivalent to human blood plasma. Cell cytotoxicity studies give an idea of the biological compatibility of the materials, whereas cell culture studies will determine the bioactivity of the material. Appropriate cells are used depending upon the application are used for this purpose.

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1.6 Summary
A comparison of the properties of metals, ceramics and polymers make it obvious that the metals are sill the preferred choice for biomedical applications. Table 1.2: Comparison of properties of different materials on a relative scale Property Strength Toughness Corrosion Resistance Biocompatibility Bioactivity Resorbability Metals High High High to medium High Medium None Ceramics Medium Low Very high High High Low Polymers Low Very Low Low Low High High

With the advancement of materials technology, new processing routes with specific property treatments are now available. This has led researchers to prepare composites of metals and ceramics that combine the best properties of both types. So to the next stage of developing of biomaterials requires the materials that are bioactive, can stimulate tissue and cellular response and remain in the body for years without any adverse effects. Composites through bulk processing or through surface treatments are being developed which give optimum properties of the constituent materials and avoid all harmful effects.

CHAPTER 2 LITERATURE SURVEY

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2.1. Introduction
Metallic biomaterials are still the preferred choice of materials for biomedical applications.Mg preferred in current research due to its excellent bio degradable property .As a lightweight metal with mechanical properties similar to natural bone, a natural ionic presence with significant functional roles in biological systems, and in vivo degradation via corrosion in the electrolytic environment of the body, magnesium-based implants have the potential to serve as biocompatible, osteoconductive, degradable implants for load-bearing applications. incorporating either intermetallics or ceramic particulates Mg alloys are generally believed to possess relatively low yield strength and poor ductility , they can be strengthened by

2.2. Advantages of Mg as biomaterial:

Magnesium implants corrode in the body with time thus eliminating the need of a second surgery for implant removal ie degradable Stress shielding can be minimised or eliminated properties similar to human bone Good mechanical properties Excellent biocompatibility Non Toxic in nature Specific strength, low density, elastic modulus Temporary implant for Hard tissue application

2.3. History of Mg alloys in biomedical application

The first use of magnesium was reported by Lambotte in 1907, who utilized a plate of pure magnesium (actual purity level unknown) with gold-plated steel nails to secure a fracture involving the bones of the lower leg (Lambotte et al., 1932) Troitskii and Tsitrin, in 1944, reported on 34 cases where magnesium, alloyed with small levels of cadmium, was fashioned into plates and screws and used to secure various fractures. Of the 34 cases, 9 were unsuccessful; these failures were attributed to infection, or difficulties arising with the mounting of a plaster cast, which presumably did not allow for treatment of gas cysts. It is
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reported that the mechanicalintegrity of most were maintained for 68 weeks, with complete resorption occurring in 1012 months (Troitskii et al., 1944).Results were reported by Znamenski in 1945, where magnesium alloy containing 10wt% aluminum was used to treat gunshot wounds in two young men. McBride reports on the use of screws, pegs, plates and bands prepared from magnesium aluminummanganese alloys to secure 20 fractures and bone grafts ( Staiger et al. 2006) . These early examples imply that magnesium-based materials are non-toxic and may actually stimulate bone tissue healing. However, the rate of corrosion of pure magnesium, or these simple alloys, occurs at a rate that is too rapid to allow sufficient time for healing as it is desirable to have the implanted fixture present for at least 12 weeks (Witte et al., 2005). In an effort to improve the corrosion resistance of magnesium, more complex alloying compositions may be necessary, with the addition of small levels (.o4%) of rare earth elements having the most significant effect. Stroganov et al. report that magnesium alloyed with 0.44 wt% rare earth metal, 0.051.2 wt% cadmium, 0.051.0wt% calcium or aluminum, and variable, traces (0.8%) levels of manganese, silver, zirconium or silicon had a slowed corrosion rate, with pins 3mm in diameter present for 5 months, and pins 8mm in diameter present for 11 months in vivo. No information was given as to how long the mechanical integrity of the implants survived or if the otentially toxic effects of the alloying elements were considered (Stroganov et al. 1972). More recently, in vivo degradation of magnesium-based alloys, comparing two alloys containing only aluminum and zinc, and two alloys with rare earth element combinations (Witte F et al,. 2005). The aluminum zinc alloys contained 3wt% aluminum and 1 wt% zinc (AZ31), and 9 wt% aluminum and 1 wt% zinc (AZ91). The first rare-earth alloy was composed of 4wt% yttrium and 3wt% of a rare earth metal mixture consisting of neodymium, cerium and dysprosium (WE43). The final rare-earth alloy consisted of 4wt% lithium, 4 wt% aluminum and 2wt% of a rare earth element mixture of cerium, lanthanum, neodymium and praseodymium (LAE442). The implants consisted of rods 1.5mm in diameter and 20mm in length, inserted into the femur of guinea pigs. A rod of polylactide of the same dimensions was used as a control. Radiographs were taken frequently, and implants were harvested at 6 and 18 weeks. Synchrotron- radiation-based microtomography was used to characterize the degradation of the implants, for which complete degradation was observed in 18 weeks (Witte et al., 2001). Significantly increased (P 0:05) bone area was observed in all groups with magnesium-based implants at weeks 6 and 18, in comparison
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to the polymer control. Increased bone area to magnesium implants. Subcutaneous gas pockets were observed after 1 week, which were removed using a syringe, and were not observed after 23 weeks. Adverse effects due to the formation of subcutaneous gas were not observed. The slowest rate of corrosion was noted for LAE442, while AZ31, AZ91 and WE43 degraded at similar rates .Energy dispersive X-ray analysis (EDX) was used to form elemental maps of the corrosion layer and new bone tissue. The rare earth elements were shown to be localized in the corrosion layer, and were not detected (Witte et al., 2005)

2.4 Requirement of alloying:

Two primary groups of magnesium-based alloys are those that contain 210 wt% aluminum with trace additions of zinc and manganese, with products that demonstrate moderate corrosion resistance and improved mechanical properties The second group uses a mixture of rare earth elements in combination with another metal such as zinc, yttrium, or silver, and a small amount of zirconium which imparts a fine grain structure and enhanced mechanical properties Alloying is an essential step to improve mechanical properties and corrosion resistance of magnesium (Li et al., 2004).Protective coatings and surface treatments can also be applied to improve the corrosion resistance and potentially improve biological compatibility and biological activity of magnesium-based implants. An appropriate alloying composition can improve the corrosion resistance, mechanical properties and the ease of manufacture of magnesium-based materials. Two primary groups of magnesium-based alloys are those that contain 210 wt% aluminum with trace additions of zinc and manganese, with products that demonstrate moderate corrosion resistance and improved mechanical properties ( Li et al, 2004)]. The second group uses a mixture of rare earth elements in combination with another metal such as zinc, yttrium, or silver, and a small amount of zirconium which imparts a fine grain structure and enhanced mechanical properties (Li et al, 2004). Little is known about the in vivo corrosion characteristics of these metals; however the work of Witte et al. suggests that some magnesium-rare earth alloys have a slightly enhanced corrosion resistance (Witte et al, 2005)

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As these materials are used in the body, care must be taken to choose alloying elements that are non-toxic. For orthopedic applications, protective coatings for magnesium must be non-toxic, and aim to improve the biocompatibility/bioactivity of the implant. Simple but effective options include alkali-heat treatments, which may induce a biomimetic precipitation of calcium phosphate at the implant surface (Li et al., 2004)

2.5 Functions of few alloying elements:

Ca- Ductility improvement also grain refiner Zr-Grain Refinement RE-High temp. properties Zn-age hardening and combination of good ductility and strength Ag addition also improves the micro-hardness La addition of 1.5% (Mg7Al1Zn1CaxLa ) improves mechanical properties by grain refinement

2.6 Porous magnesium microstructures:

A microstructure of interconnected pores in the solid matrix results in an implant with lower density and considerably altered mechanical properties and deformation behavior . Appropriate choice of pore size can result in significantly improved integration of the implant with natural tissue. The number, size, shape and connectivity of pores have significant effects on the Youngs modulus and yield stress (Gibson et al.,1988). Cancellous bone is an interconnected, porous structure, and consequentially it exhibits deformation mechanisms that are typical of such materials (Grey et al, 2002). The porosity of cancellous bone varies considerably from 30% to 95%, which significantly influences the resulting mechanical propertiesyield stresses of cancellous bone are reported to range from 3 to 20MPa, with corresponding Youngs moduli from 10 to 40 GPa . Thus, the inclusion of porosity can benefit the material by increasing biological integration and by adjusting the mechanical properties to comply with the natural bone system at hand. On the other hand, too many large pores will compromise the mechanical properties of the implant making it unsuitable for a load bearing applications. (Gibson et al,.1988)
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2.7 Different processing routes:

A number of techniques are available to produce an open porous metallic structure, however to ensure the biocompatibility of the resulting material, one must be careful when selecting the reagents or methods used for the generation of the pores. Porous magnesium metal implants suitable for biomaterial applications have been prepared using argon gas injection to molten magnesium (Banhart et al, 2001) a plaster casting method using a polyurethane foam, a powder metallurgy techniques using space-holding particles. While plaster casting and powder processing using space-holding particles produced viable products, production using foaming by injection of argon gas to the molten metal did not produce a product with consistent morphology and this approach has been abandoned (Banhart et al , 2001) . Pore volume and size significantly affect the mechanical properties of porous magnesium materials. In all cases, the inclusion of porosity results in a material with reduced yield strength and modulus, corresponding with the lower range of mechanical properties of natural bone (Yamada et al,.2000)

2.8. Ball Milling of Mg:

Mg nanocomposite has been successfully synthesized through mechanochemical milling. Nanoparticles embedded in the Mg matrix have been observed. The grain size of the milled specimen is about 77 nm (Leu et al. 2004) The influence of the high-energy ball milling on the corrosion behavior of magnesium in aqueous media has been investigated through electrochemical experiments complemented by morphological, structural, chemical and surface analyses.Polarization curves show that the milling procedure improves the magnesium corrosion resistance in passive conditions (KOH solution) and in more active corrosion conditions (borate solution). This is illustrated by the corrosion potential which becomes nobler with milling. The variation of the polarization resistance and related corrosion current with milling time is also an indication of the improvement of the Mg corrosion resistance due to the milling. The Mg crystallite size is reduced from >100 to 34 nm after 10 h of milling and does not decrease significantly with further milling (M.-H. Grosjean et al.2004). The MMCs were produced by mixing 20wt% of HA powder and the balance of powdered magnesium alloy AZ91D into an AZ31 can of 70mm diamete r (30 micron). MMC made of magnesium alloy AZ91D as a matrix and hydroxyapatite (HA) particles as reinforcements have been investigated in vitro for mechanical, corrosive and cytocompatible properties. Magnesium alloy-based metal
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matrix composites are cytocompatible biomaterials with adjustable mechanical and corrosive properties. The distribution and size of the HA particles are of major importance for mechanical and corrosive properties. Corrosion tests revealed that HA particles stabilised the corrosion rate and exhibited more uniform corrosion attack in artificial sea water and cell solutions (Witte et al. 2007)

CHAPTER 3 SCOPE AND OBJECTIVES

As established earlier magnesium is the preferred material for biodegradable application due to its superior properties. Magnesium has shown bioactivity better than most of the metals. It provides the desired strength and has modulus closest to human bone, thus making it a good material for
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hard tissue applications. Hydroxyapatite coatings or surface additions can be done successfully done on magnesium matrix as HA integrates well with the bone tissue (good osseo-integrity) and can act as surface for cell growth. Mechanical milling is simple and convenient technique which is commercially scalable to produce nano particles. Nano particles are desired for their superior mechanical and biological properties. Nature has an affinity for nano materials and human bone tissue is also made up of nano fibers. The present research work thus aims to improve the properties of magnesium based implants by forming composites of Magnesium and HA The main objectives of the present research work can be summarized as below: 1. To prepare composites of magnesium and nano-hydroxyapatite through ball milling. 2. Compacting and sintering in conventional vacuum sintering and spark plasm sintering 3. Characterizing the composite for the metallurgical properties and mechanical properties

CHAPTER 3 EXPERIMENTAL PROCEDURES:

3.1 Materials
Magnesium : Commercially available Mganesium powders purchased from Sigma Aldrich was used. The purity was 99.9% and the size <150 m. Hydroxyapatite: HA was prepared in the lab from egg shell. hydroxyapatite is of 99.5% pure, particle size <6 nm.

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3.2 Mechanical milling:

Mechanical milling is a well established technique to obtain uniform nano powders and alloys. Milling involves cold welding, fracturing and rewelding of powder particles due to high impact of the balls. Milling was carried out in planetary ball mill (Fritsch, PM400, and Germany). Experimental details are as follows: Composition: Pure Mg, and Mg with 8, 10, 15 wt. % HA. Grinding Media: WC balls with ball to powder ratio of 20:1. Wetting medium: Ethanol Milling time: Milling time was 20 hour with cycles of 60 minutes each for milling and 15 minutes for idle time. Rotation speed was 200 rpm. Mg and HA ball milling process with the following process composition. Mg-8wt.% HA, Mg10wt.% HA, Mg-15wt.% HA compositions are milled in ethanol medium with BPR 20:1and milling media is WC. The ball-milled powders have been collected and dried in oven at the temperature of 80C for 15hours.

3.3 Compaction and vacuum Sealing:

The analyzed powders Compacted in the hydraulic press 10 Tonnage (Auto) and the process details as follows. : Die diameter: 9mm Load: 5Tonnage Time: 30secs Powder qty: 0.5gram Then the green compact is vacuum sealed for the sintering process, since Mg is highly reactive in nature. The compact is kept inside the quartz tube (12mm) and vacuum sealed (10-3mbar).
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3.4 Sintering:
Sintering is carried out in Box furnace Temperature: 70010C Soaking Time: 2Hours Heating Rate: 5 C

3.5 Spark plasma sintering:

Milled powders have also been Spark Plasma sintered to get the good compacted and sintered product. Sintering Temperature: 400 C Pressure: 50 Mpa Sintering Time: 10 minutes

3.6 Characterization: 3.6.1XRD:

The X-ray diffraction studies on powder (ball milled) and sintered samples were done with Bruker Discover D8 Diffractometer, Germany. The diffraction pattern were obtained using Cu K radiation with the scan rate of 0.15 sec per step and scan range of 20 to 80.

3.6.2 Scanning electron microscopy:

Scanning Electron Microscope (FEI Quanta 400, Holland, and FEG SEM) was performed on the powdered samples to study the morphology. Elemental analysis (EDAX) was done to assess any new phase formed or contamination occurring during ball milling and to access the distribution of Mg and HA particles.

3.6.3 Transmission electron microscopy

The ball milled powders were dispersed in ethanol and subjected to ultrasonic treatment and then observed in TEM using a carbon coated copper grid. Selected area dispersion (SAD) patters were obtained.

3.6.4. Hardness testing:

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Vickers Hardness measurement was done using micro-indenter. The the load applied was 100g for 15 seconds. Hardness was measured at 8 different points and the average calculated.

CHAPTER 4 RESULTS AND DISCUSSIONS 4.1 Introduction

This chapter deals with the processing of commercially available magnesium through mechanical milling and characterization of the milled product. The milled magnesium was characterized using X-ray diffraction (XRD) method, Scanning Electron Microscopy (SEM), Transmission Electron Microscope (TEM), evaluation of hardness.

4.2 Starting material characterization

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Fig 4.1 SEM image of as received pure magnesium powder and corresponding EDAX analysis

Electron microscope images (Fig 4.1) of the powders confirm that the starting materials purity and particle size before the processing. And also from the SEM images we found that magnesium powders has got very little oxidation also, which is unavoidable in Pure Magnesium. Images confirms that its of <150 particle size and from the bright field TEM image of the hydroxyapatite powder (Fig 4.2) shows that it is less than 100nm size.

Fig 4.2 Bright field TEM image of lab prepared nano hydroxyapatite powder

4.3 X-ray diffraction:

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Fig 4.3 XRD of nanohydroxyapatite at room temperature and heated to 850C.

From the Fig 4.3, its evident that HA was not decomposed at 850C and also coarsening of HA particles observed. The fraction of crystallinity was increased after heating. Mechanical Milling of commercially available magnesium powder was done for 20 hours. The apparatus was allowed to cool for 60 minutes after every 15 minute milling cycle to avoid excessive heating.

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Fig 4.4 XRD patterns of ball milled powders

The X-ray diffraction studies on powder (ball milled) and sintered samples were done with Bruker Discover D8 Diffractometer, Germany. The diffraction pattern were obtained using Cu K radiation with the scan rate of 0.15 sec per step and scan range of 20 to 80. The XRD pattern of ball milled powders of various composition of Mg-HA (Fig 4.4) showed that there is no new phase and contamination in the powders. Instead, there is a shifting of high intensity peak due to the micro strain in the milling powders. It confirms that there is no contamination due to tungsten carbide milling media,since we are not observing any W or C peaks in the pattern. Fig 4.5 shows the powders after ball milling. The powder was flattened and flakes were produced after 20h of milling.

4.4 SEM Images of Ball milled powders:

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Fig 4.5 SEM images of ball milled powders

4.5 TEM analysis of Ball Milled powders:

The ball milled powders were dispersed in ethanol and subjected to ultrasonic treatment and then observed in TEM using a carbon coated cupper grid. Selected area dispersion (SAD) patters were obtained. Transmission electron micrographs shows that the after high energy ball milling process powders are still remains as crystalline. The HA ceramic particles uniformly coated/deposited over the Mg particle. Very well distributed particles observed in the TEM bright field imaging. Fig 4.6 of Bright filed imaging shows very good coating/depositing of HA particle over Mg particle. Fig 4.7 shows the HA covered on the Mg particle after ball milling.

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Fig 4.6 Bright field TEM image and corresponding SAED pattern of ball milled powders a) pure Mg, b) Mg-8HA, c) Mg-10HA and d) Mg-15HA

Fig 4.7 Bright field TEM image of nano-HA powder covered on magnesium particle after 20h of ball milling

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Fig 4.8 Vacuum sealed compact for sintering

4.6 SEM images of sintered samples

Fig 4.9 SEM observations of sintered pure magnesium in as received condition: a) BSE image, b) corresponding EDAX analysis within the grain and c) at the grain boundary.

Fig 4.8 shows the samples vacuum sealed in quartz tubes. Fig 4.9 shows that typical Mg powders sintering characteristics. Grain coarsening during the sintering was observed. But as expected oxidation is inevitable during the sintering process which is clearly demonstrated In the EDAX analysis in the Scanning Electron microscope. The grain was pure magnesium and the grain boundary was got oxidized.

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Fig 4.10 SEM observations of sintered Mg-8HA composite: a) BSE image, b) corresponding magnified image, c) EDAX analysis within the grain and d) at the grain boundary.

Sintering of 20 hours ball milled Mg-8%HA powders has got sintered well and HA particles are seen along the grain boundaries of the Mg particle. And also due to the presence of the HA the Mg particle might have got oxidized. EDAX analysis shows that the major part is magnesium particle and Ca, p and few amount of oxide as well (Fig 4.10).

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Fig 4.11 SEM observations of sintered Mg-10HA composite: a) BSE image, b) corresponding magnified image, c) EDAX analysis within the grain and d) at the grain boundary.

SEM analysis of sintered Mg-10%HA shows that particle got sintered and coarsening of the particle after the process. Mg particle might have got flattened during the milling process and HA particles got embedded over the flat and elongated Mg particles. In this composition HA particle tend to settle at the grain boundaries of the Mg particles , and during sintering they formed MgO also. EDAX results of the sintered pellet also confirm the same effect of oxidation which may from HA particle presence (Fig 4.11).

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Fig 4.12 SEM observations of sintered Mg-15HA composite: a) BSE image, b) corresponding magnified image, c) EDAX analysis within the grain and d) at the grain boundary.

SEM images of Mg-15%HA ball milled and sintered powders showed that three different phases such as Mg,MgO,HA presence which is evident by presence of Ca,P. Due to milling and compaction process the HA particle got embedded on the grain boundary of the mg particles. During sintering process the HA particle reacted with the mg which results in formation of MgO. From the BSE images shows that clear distinct phases in the sintered part and EDAX analysis within the grain and at the grain boundary confirms the presence of MgO in the sintered part (Fig 4.12).

4.7 XRD analysis of Vacuum sintered samples:

XRD analysis of the sintered pellet ( Fig 4.13) showed that the corresponding peaks of Mg,HA and also MgO.It does not shows any indications of new elements which confirms that there is no d4issociation of hydroxyapatite as such. Peaks of Mg at 2 value 35.5 and most of the
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corresponding Mg peaks. HA peaks also observed at the respective 2 values for the presence too. Higher the composition of the HA intensity of the MgO also intensifies shows that HA may the source for the oxidation when its going to the higher temperature which is the sintering temperature of the Mg-HA composite.

hydroxyapatite. MgO peaks also shows at 2 values of 42 and 62 respectively, indicates its

Fig 4.13 XRD pattern of the sintered pellet

4.8 Micro Vickers hardness measurement

In the figures 4.14 & 4.15, Vickers hardness distribution in the sintered sample and Hardness average of the samples is indicated. From the distribution graph, we observe that higher the concentration of HA it reduces the hardness of the sintered sample due to its ability to observe the energy which is applied in ball milling process and also due to its affects the size reduction capabilities of ball milling results in lesser hardness in higher HA content samples.
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Fig 4.14 Micro hardness distribution across the radius of the sintered pellets

Fig 4.15 Average hardness of the sintered pellets

4.10 Spark plasma sintering

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Fig 4.16 photographs of the spark plasma sintered samples

4.11 XRD Pattern of SPS sample

Fig 4.16 shows the spark plasma sintered pellets. XRD analysis of the SPS pellets (Fig 4.17) showed that the corresponding peaks of Mg,HA and also MgO. It does not shows any indications of new elements which confirms that there is no dissociation of hydroxyapatite as such. It does not contain carbon peaks also which confirms the that there is no carbon entrapment into the sps sample even though the SPS die is made up off graphite. We observed Mg at 2 value 34.5 and most of the corresponding Mg peaks. HA peaks also observed at the respective 2 values for the hydroxyapatite. MgO peaks also shows at 2 values of 42 and 62 respectively, indicates its presence too. Higher the composition of the HA intensity of the MgO also intensifies shows that HA may the source for the oxidation when its going to the higher temperature which is the sintering temperature of the Mg-HA composite. Here the intensity of the MgO peak is largely reduced due the controlled compaction and sintering under the sophisticated equipment used and hardly any chance of atmospheric contact.

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Fig 4.17 XRD patterns of the SPS samples

4.12 SEM Analysis of SPS samples

Fig 4.18 shows the SEM and EDAX observations of the composites. The grains were observed as flakes in morphology. The grain boundary was observed to be with different contrast as observed in back scattered electron image. Corresponding EDAX analysis confirms the presence of oxygen, calcium and phosphorous elements along with magnesium. The results suggest that the grain boundary consist MgO and hydroxyapatite which was also confirmed by XRD.

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Fig 4.18 SEM observations of SPS composites:

4.13 Vickers micro-hardness SPS samples

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Fig 4.19 Microhardness distribution across the radius of the sintered pellets

In the figures 4.19 & 4.20, Vickers hardness distribution in the sintered sample and average hardness of the samples are indicated. From the distribution graph, we observe that higher the concentration of HA it reduces the hardness of the sintered sample due to its ability to observe the energy which is applied in ball milling process and also due to its affects the size reduction capabilities of ball milling results in lesser hardness in higher HA content samples. In 10%HA composite has got good hardness due to the HA got into the Mg grain boundary and forms MgO which may might be the reason for higher hardness. Hardness also has got variation in over the distance due to various reasons like porosity level, go presence ,composite structure formation during ball milling.

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Fig 4.20 Average hardness of the sintered pellets

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CONCLUSIONS
Magnesium hydroxyapatite (8%,10% and 15% HA) composites were synthesized by ball milling and consolidated by conventional vacuum sintering and spark plasma sintering The ball milled powders were slightly oxidized after 30h of ball milling and the covering of nanoHA over the magnesium particles was observed in all the compositions Presence of magnesium oxide and Ca/P observed at the grain boundary of sintered compacts in both the cases suggest the oxidation of the sintered powders and the presence of HA after sintering. The microhardness studies of the samples indicate improvement in hardness for the samples compared to pure magnesium sintered in both routes As the powder deformed into fine flakes due to ball milling, the microstructure of the composites clearly shows the combination of magnesium oxide and HA at the layers like grain boundary.

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SCOPE FOR THE FUTURE WORK

1. The influence of secondary phase particles in controlling the biodegradation and the role of

magnesium oxide and HA on the bioactivity of the composite in physiological environment can be assessed by in-vitro bioactivity studies

2. The level of toxicity of the new developed composites on the cell viability can be found by biocompatibility studies

3. The cell attachment and growth kinetics on the composites can be investigated by doing cell adhesion studies to find the cells response to the new developed magnesium based degradable composites

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