Advanced Biomaterials and

Manufacturing Process

Siddhartha Maiti
 What is a Biomaterial?
 A material intented to interface with biological systems to evaluate,
treat, augment or replace any tissue, organ or function of the body.
 Biomaterial is used to make devices to replace a part or a function of
the body in a safe, reliable, economic and physiologically acceptable
manner.
 Materials of synthetic as well as of natural origin in contact with tissue,
blood, and biological fluids, and intended for use for prosthetic,
diagnostic, therapeutic, and storage applications without adversely
affecting the living organism and its components.
 The major difference of biomaterials from other classes of materials is
their ability to remain in a biological environment without damaging the
surroundings and without getting damaged in that process.
 Therefore, biomaterials require both biological and materials properties
to suit a specific application.
 Need for Biomaterials
 Millions of patients suffer end stage organ and tissue failure
annually.
 $400 billion annually for treatment.
 8 million surgical procedures.
 Treatment options include transplantation, reconstruction,
mechanical devices.
 The options and limitations
 Transplantation
 Critical donor shortage ▪ 3000 livers annually for 30000 people in
need
 Disease Transmission ▪ HIV ▪ Hepatitis ▪ Other transmittable
diseases.

 Surgical Reconstruction  Not always possible  Complications

 Mechanical Devices
 Engineering approach – engineer new tissues, systems
 Limited by ▪ Complexity of the human body ▪ Multiple functions ▪
Living components versus non living components
▪ Materials? ▪ Tissue based ▪ Polymers ▪ Metals ▪ Ceramics
Biomaterials involved in Human Body
 Polymeric Biomaterials: Adv & Disadv
• Easy to make complicated • Leachable compounds
items • Absorb water & proteins
• Tailorable physical & etc.
mechanical properties • Surface contamination
• Surface modification • Wear & breakdown
• Immobilize cell etc. • Biodegradation
• Biodegradable • Difficult to sterilize
• Examples of Polymeric Biomaterials:
• PMMA
• PVC
• PLA/PGA
• PE
• PP
• PA
• PTFE
• PET
• PUR
• Silicones
 Bioceramic: Advantages and disadvantage
• High compression strength • High modulus
• Wear & corrosion (mismatched with bone)
resistance • Low strength in tension
• Can be highly polished • Low fracture toughness
• Bioactive/inert • Difficult to fabricate

• Examples of Bioceramic:
• Alumina
• Zirconia (partially stabilized)
• Silicate glass
• Calcium phosphate (apatite)
• Calcium carbonate
 Metallic Biomaterials:Advantages &
Disadvantages
• High strength • High moduls
• Fatigue resistance • Corrosion
• Wear resistance • Metal ion sensitivity and
• toxicity
Easy fabrication
• Metallic looking
• Easy to sterilize
• Shape memory

• Examples of Metallic Biomaterials:

• Stainless steel (316L)
• Co-Cr alloys
• Ti6Al4V
• Au-Ag-Cu-Pd alloys
• Amalgam (AgSnCuZnHg)
• Ni-Ti
• Titanium
Materials for Use in the Body
 Some application of biomaterials
Application Types of Materials
• Skeletel system
• Titanium , Stainless steel
Joint replacement(Hip, knee)
• Stainless
Bone platesteel, Co-Cr alloy
• PMMA
Bone cement
• Hydroxylapatie
Artificial tendonTeflon, Dacron
and ligment
• Titanium, alumina, calcium phosphate
Dental implant
• Cardiovascalar sysem
•• Dacron, Teflon,
Blood vessel Polyurethane
prosthesis
•• Reprocessed
Heart valve tissue, Stainless steel, Carbon
•• Silicone
Catheter rubber, teflon, polyurethane
• Organs
•• Polyurethane
Artificial heart
•• Silicone-collage composite
Skin repair template
•• Cellulose, polyacrylonitrile
Artificial kidney
•• Silicone rubber
Heart-lung machine
• Senses
• Platium electrodes
• Cochlear replacement
• PMMA, Silicone rubber, hydrogel
• Intraocular lens
• Silicone-acrylate. Hydrogel
• Contact lens
• Collagen, hydrogel
• Corneal bandage
 Significance of Biomaterials
 Replace diseased part – Dialysis
 Assist in Healing – Sutures
 Improves Function – Contacts
 Correct Function – Spinal rods
 Correct Cosmetic – Nose, Ear
 Aids – Probes / Catheters
 Replace rotten – Amalgam
 Replace dead - Skin
 Evolution of Biomaterial Science & Technology
• 1st generation (since 1950s)
Goal: Bioinertness
• 2nd generation (since 1980s)
Goal: Bioactivity
• 3rd generation (since 2000s)
Goal: Regenerate functional tissue
 First generation Implants
 ‘Ad-hoc’ [unplanned] Implants
 Specified by physicians using common and borrowed materials
 Most successes were accidental rather than by design
 Examples – First generation Implants
 Gold fillings, wooden teeth, PMMA dental prosthesis  Steel, gold,
ivory, bone plates etc.  Glass eyes and other body parts

Vascular Grafts
 Second generation Implants
 Engineered implants using common and borrowed materials
 Developed through collaborations of physicians and engineers
 Built on first generation experiences
 Used advances in materials science
 Examples – Second generation implants
 Titanium alloy dental and orthopaedic implants  Cobalt-chromium
implants  UHMW polyethylene bearing surfaces for total joint
replacements  Heart valves and Pacemakers.

Artificial hip joint

 Third generation Implants
 Bioengineered implants using bioengineered materials
 Few examples on the market
 Some modified and new polymeric devices
 Many under development
 Example – Third generation implants
 Tissue engineered implants designed to re-grow rather than replace
tissues  Some resorbable bone repair cements  Genetically
engineered ‘biological’ components.

Biological valve
 Characteristics of Biomaterials
 Physical Requirements
• Hard Materials. • Flexible Material.

 Chemical Requirements
• Must not react with any tissue in the body. • Must be non-toxic to
the body. • Long-term replacement must not be biodegradable.
 Main features of Biomaterials for medical
applications
 Biofunctionality
• Playing a specific function in physical and mechanical terms

 Biocompatibility
• The ability of a material to perform with an appropriate host response in a
specific application.
• Absence of carcinogenicity (the ability or tendency to produce cancer)
• Absence of immunogenicity (absence of a recognition of an external factor
which could create rejection)
• Absence of teratogenicity (ability to cause birth defects)
• Absence of toxicity
• Resistance to blood clotting (eg. Hemodialysis membrane- in contact wit the
patient for 3 hrs)
• Resistance to bacterial colonization (eg-urinanry catheter-may be inserted
for a week)
• Normal, uncomplicated healing (eg.- tissue engineering applications)
 Host Response
 In order to develop new materials, it is desirable to understand the in vivo
host response of various biomaterials.
 Ideally, biomaterials should not induce any change or provoke undesired
reaction in the neighboring or distant tissues.
 An important aspect of host response involves the formation of a structural
and biological bond between the material and host tissues.
 When the biocompatibility is lacking, materials cause tissue reactions,
which may be systemic or local.
 Systemic responses can be toxic or allergic and triggered by the products of
metallic corrosion and polymer degradation, release of micro particles from
materials, and the presence of contaminants.
 Different human systems (such as respiratory, circulation, or digestive)
respond in different ways to contact with foreign bodies or materials.
 Host Response
 Depending on the biocompatibility and host reaction, biomaterials can be
broadly classified into three main categories on the basis of various types of
host responses of biomaterials after implantation into the living body:
 a) Bioinert / biotolerant: Bioinert materials are biocompatible materials,
but cannot induce any interfacial biological bond between implants and
bone.
 b) Bioactive: Bioactive materials are a group of biocompatible materials
that can attach directly with body tissues and form chemical and biological
bonds during early stages of the post implantation period.
 c) Bioresorbable: Bioresorable materials are the type of biocompatible
materials that are gradually resorbed before they finally disappear and are
totally replaced by new tissues in vivo.
 Host Response
 When a bioinert material is implanted, a capsule - like layer forms on the
surface of the implant to keep it isolated from the living part of the body. For
example, bioinert ceramics, such as alumina or zirconia, develop
fibrous capsules at their interface when implanted.
 However, it is important to note that the thickness of an interfacial fibrous
layer depends upon motion and the extent of required fit at the interface.
Therefore, a bioinert material is not useful for long-term application.
 The most significant class of biomaterial is bioactive material, which can
potentially behave as the part of a living body.
 A few examples of bioactive materials are 45S5 bioglass and calcium
phosphates (HA). For bioactive materials, the interfacial bond prevents
motion between the implant – tissue interfaces and imitates the type of
interface found when natural tissues repair themselves.
 The third kind of material is bioresorable or degradable, which degrades
with time inside the body’s environment. The degradation rate should be
such that the regeneration rate of new tissue will be same as the material
resorption rate. Tricalcium Phosphate (TCP) and bone cement are the
two examples of bioresorable materials.
 CELL – MATERIAL INTERACTIONS
 The interaction between biomaterials and natural tissues is an important
scientific issue and understanding this issue is essential to designing new
biocompatible materials.
 In understanding the interaction and integration of biomaterials in a human
body, it is worthwhile to mention the physicochemical conditions of the
human body ’ s environment.
 For example, nominal pH values vary over a wide range of 1.0 (gastric
content) to 7.4 (blood). Additionally, pH values can change depending on
health conditions (disease, etc.).
 The temperature of the normal core of human body is around 37.4°C;
however, deviations over a range of temperature 20 – 42.5°C have been
reported for diseased patients.
 As far as the inorganic composition of the human body is concerned, total
body burden of Ca, Na, Cl ions is much higher and also traces of Mg, Fe,
Zn, Cu, Al, and so on, are present in cytoplasm.
 CELL – MATERIAL INTERACTIONS
 Once a material is implanted in an
animal, a large number of protein
molecules are adsorbed on biomaterial
surface. This is because of the
abundance of protein as an order of 109
number of protein molecules per
eukaryotic cell is estimated in the human
body.
 Protein adsorption on an implant takes
place first because of faster adsorption
kinetics, and this acts as precursor to the
cell – material interaction.
 The initial interaction is established
through the interaction of cell surface
receptors with adsorbed protein ligands.
Such protein - to - protein binding
importantly helps a cell to spread on the
material surface.
 CELL–MATERIAL INTERACTIONS
 After implantation of a biomaterial, a
monolayer of protein molecules gets absorbed
on the material surface within a minute.
 Subsequently, the transport of various cell
types towards the biomaterial surface occurs
and the interaction of the integrin proteins of
cell surface with the absorbed protein of
biomaterials surface is established.
 The secretion of cell enzymes of various cell
type form an extra cellular matrix (ECM).
Depending on cell biomaterial interaction,
various cell types can adhere in a self -
organized manner to form a tissue.
 To this end, an important event is the
formation of the small blood vessels
(angiogenesis) as well as formation of large
blood vessels (vasculogenesis) within the
newly - formed tissue layer. Such formation is
necessary for the supply of nutrients locally to
various cell types as well as for the removal of
waste from ECM.
 EXPERIMENTAL EVALUATION OF
BIOCOMPATIBILITY
 Any research program of assessment on biomaterials must include a range
of in vitro and in vivo tests, as stated by various standard agencies (for
example, International Organization for Standardization [ISO]).
 In vitro tests are lab scale simulated experiments, which are rapid and are a
must as initial screening tests. From the results of the in vitro tests, one
cannot obtain any information of inflammation and immune response of the
materials.
 Also, most of the in vitro experiments use single cell lines, which do not
reflect the actual tissue interaction (involving multiple cell types) in vivo .
 Although the in vitro experiments are inexpensive, such tests do not provide
appropriate representation of physiological conditions. These tests,
nevertheless, are effective as the first step of biocompatibility evaluations.
 On the other hand, in vivo experiments produce a better approximation to
the human environment. Here, the material comes in contact with different
cell types and the effect of hormonal factors can be analyzed.
 Also, in vivo tests provide interactions with extracellular matrix, blood -
borne cells, protein and molecules. These experiments can be regarded as
the second step prior to clinical use.
 EXPERIMENTAL EVALUATION OF
BIOCOMPATIBILITY: In Vitro Tests
 Cytotoxicity: The cytotoxicity experiments determine whether the material
is toxic in contact with some particular cell lines.
 As far as the experimental evaluation of biocompatibility is concerned, the
cytotoxicity tests are widely cited as the primary assessment of
biocompatibility
 As a first step, the sterilization of the samples is carried out in order to
remove other micro-organisms, if present on the surface.
 Depending on type of implant materials (i.e., chemistry and chemical
composition), the sterilization is either carried out in steam autoclave (15
psi, 121°C, 20 minutes) or using γ - ray irradiation.
 MTT assay is a standard colorimetric assay (an assay which measures
changes in color) to quantify cellular proliferation (cell growth).
 It is used to determine cytotoxicity of potential medicinal agents and other
toxic materials. Yellow MTT (3-(4,5 - Dimethylthiazol - 2 - yl) - 2,5
diphenyltetrazolium bromide, a tetrazole) is reduced to purple formazan
in the mitochondria of living cells.
 EXPERIMENTAL EVALUATION OF
BIOCOMPATIBILITY: In Vitro Tests
 A solubilization solution (usually
either dimethyl sulfoxide or a
solution of the detergent sodium
dodecyl sulfate in dilute
hydrochloric acid) is added to
dissolve the insoluble purple
formazan product into a colored
solution.
 The absorbance of this colored
solution can be quantified by
measuring at a certain
wavelength (usually between
500 and 600 nm) by a
spectrophotometer.
 This reduction takes place only
when mitochondrial reductase
enzymes are active, and
therefore, conversion is directly
related to the number of viable
(living) cells.
 EXPERIMENTAL EVALUATION OF
BIOCOMPATIBILITY: In Vitro Tests
 Genotoxicity: In this in vitro experiment, it is primarily observed whether
any genetic mutation occurs in the cells in direct contact with the
biomaterial surface.

 Hemocompatibility: Hemocompatibility evaluates the material’s

compatibility with red blood cells. In particular, thrombogenic property or
changes in RBC content in a blood stream flowing over the biomaterials are
assessed and a better thrombus material should ideally show limited
thrombus formation. Such evaluation is a must for cardiovascular implant
materials.
 The examples of hemocompatibile materials are PolyTetra Flouro Ethylene
(PTFE), Diamond Like Carbon (DLC), and so on.
 EXPERIMENTAL EVALUATION OF
BIOCOMPATIBILITY: In Vivo Tests
 Sensitization: This is an in vivo test, where materials are kept in the
subcutaneous region of an animal and the observations are the change in
skin color, allergic effect, or some other irritations.

 Implantation: Implantation is an in vivo experiment, where a sample of a

predefined shape is placed in the long bone of a mammal (rabbit, rat and
mouse).
 After a specified time frame, the samples and surrounding tissues are
examined histopathologically to determine the in vivo response of the
materials.

 Carcinogenicity: Carcinogenicity is a long term in vivo experiment, which

determines any cancerous effect on the cells in contact with materials. The
examples of carcinogenic materials are Pb, Sn, and so on.
 EXPERIMENTAL EVALUATION OF
BIOCOMPATIBILITY

 ISO (international standard organization) draft categorizes the devices by

the nature of their contact with the body:

 a)Surface - containing devices, such as electrodes, compression

bandages, contact lenses, urinary catheters, and so on.
 b)External communicating devices, such as dental cements,
arthroscopes, intravascular catheters, dialysis tubing, and so on.
 c) Implant devices, such as hip and knee prosthesis, pacemakers, artificial
tendons, heart valves, and so on.
 EXPERIMENTAL EVALUATION OF
BIOCOMPATIBILITY: Other Tests
 Sufficient knowledge about the biodegradation of biomaterials is essential in
the evaluation of local or systemic effects, which can be caused in patients.
 For example, the corrosion of orthopedic alloys causes the release of
various metal elements in human tissue and has to be assessed with
respect to levels, kinetics, and chemical state of the ions.
 Other important tests for load - bearing implants include friction and
wear tests. Besides evaluation of frictional and wear resistance properties,
the wear debris particles need to be analyzed in terms of size/ size
distribution and chemistry.
 In addition, in vitro dissolution tests assess the weight change or
deposition of any mineralized phase (such as CaP - rich) on the surface of
biomaterial.
 STEPS FOR CHARACTERIZATIONS OF
BIOMATERIALS
 The first step is the material development and its characterization. Previous
experience and literatures help in choosing the appropriate system to
evaluate a material for a specific application.
 After evaluating the physical properties and some other necessary tests, the
material goes to microbiology section, where the material is sterilized by
ethanol or gamma ray or some other techniques.
 Depending on the desired biomedical application of the materials under
investigation, it goes to thrombosis lab and tissue culture lab, to evaluate the
in vitro properties of materials.
 Subsequently, material goes to the in vivo toxicity lab. Here, materials extract
is injected into animal bodies or material is placed in the animal body.
 In the histopathology lab, animal tissues are prepared for microscopic
analysis. Special techniques are adopted to make the sample for optical,
scanning and transmission electron microscopy.
 STEPS FOR CHARACTERIZATIONS OF
BIOMATERIALS
 There are some important aspects to choose the animal for in vivo
experiments. The animal welfare committee and ethical committee decide
the number of animals and the type of animals for particular in vivo
experiments. Some guidelines need to be followed.

 When the material is successfully selected for a particular application, it

goes to implant biology section to shape the material into final use.

 The other important aspects of biocompatibility testing are clean room

practice and microbiological evaluation of materials.
 METALS AND ALLOYS IN BIOMEDICAL
APPLICATIONS
 Because of better physical properties such as strength, toughness, or
ductility, some biocompatible metals and their alloys are often used for joint
and bone implants.
 The applications of metals include bone replacement, bone repair, metal
plates for fractures, dental implants (fillings and posts), screws and
staples, parts of other devices like artificial hearts, pacemakers and
catheters.

 The major issues for metallic implants are biocompatibility, wear resistance
and corrosion resistance in body fluid.
Application of metallic biomaterials as implants in different areas of the human body
SEM images, revealing the adherence of L929 fibroblast cells seeded for 24
hours on HDPE - 20HAp - 20Al (S4) (a) &(b), HDPE - 40HAp (S6) sample (c)
&(d)
 Issues Limiting Performance of Metallic
Biomaterials
 Wear of Implants: For implant applications, wear
of metallic implants is a serious concern for
various biomaterials.
 The examples are different joints of the human
body, where two similar or dissimilar materials
come in contact.
 In a typical HIP prosthesis, a metallic stint is
attached to a ceramic ball, and the ceramic ball
moves inside the polymeric acetabular cup.
 At the joint of the metal–ceramic interface, fretting
fatigue could be responsible for loosening of the
implant.
 On the other hand, the ceramic ball – polymer cup
interface experiences sliding wear. The presence
of body fluid and different types of proteins may
trigger the wear rate in vivo.
 Issues Limiting Performance of Metallic
Biomaterials
 Corrosion of Metallic Implants: Most of the metallic implants corrode in
contact with body fluid.
 Sometimes, these corrosion products are harmful to the human body and in
most of the cases, the mammalian cells cannot metabolize these corroded
waste.
 Therefore, it is important to study the corrosion behavior of metallic implants
in vitro, that is, prior to in vivo application.
 Corrosion is an important factor in the design and selection of metals and
alloys for service in vivo.
 Allergenic, toxic/cytotoxic or carcinogenic (e.g., Ni, Co, Cr, V, Al) species
may be released to the body during corrosion processes.
 In addition, various corrosion mechanisms can lead to implant loosening and
failure. Therefore, biomaterials are often required to be tested for corrosion
and/or solubility before they are approved by regulatory organizations.
 Issues Limiting Performance of Metallic
Biomaterials
 Metallosis is a local destruction of soft and hard tissues due to metal
implants. It is often associated with significant osteolysis and is sometimes
reflected by an infiltration of periprosthetic soft tissues and bone by metallic
debris resulting from wear of joint arthroplasties.
 By themselves, metal ions lack the structural complexity required to
challenge the immune system. However, when combined with proteins, such
as those available in the skin, connective tissues and blood, a wide variety
of metals induce immune responses and thus should be considered harmful.
 Cobalt, chromium and nickel are the most common examples, nickel having
the strongest effect. Delayed type IV hypersensitivity is the most typical
response of a metal-sensitized individual to such metal ions.
 Corrosion of Metallic Implants
 Temperature and pH are two important factors affecting the corrosion
behavior of materials.
 Under normal conditions, body fluids have a temperature of 37◦C. This can
be regarded as a constant temperature throughout the lifespan of an implant
with respect to corrosion.
 The hydrogen evolution reaction (HER) and oxygen evolution reaction
(OER) are two important reduction reactions in corrosion in general, and
also in vivo.
 The HER in acid solutions is 2H+ + 2e− → H2,
 whereas in alkaline solutions it is written as 2H2O + 2e− → H2 + 2(OH) −
 The OER in neutral or acidic solutions can be written as O2 + 4H+ + 4e− →
2H2O, whereas in alkaline solutions it is 4(OH) − → O2 + 4e− + 2H2O
 Corrosion of Metallic Implants
 The corrosion of metallic implants can affect the surrounding tissues in three
ways: (i) electrical current may affect the behavior of cells, (ii) the corrosion
process may change the chemical environment, and (iii) metal ions may
affect cellular metabolism.
 One of the issues that arise from the release of corrosion products into the
body is systemic and remote effects.
 Mild corrosion in many cases can also produce symptoms, which range from
a local tenderness at the site of the corroded area to acute pain, reddening
and swelling over the whole general area around the device.
 Systemic reaction to metallic implants stem from this flux of metal ions
released by corrosion.
 Organ specific accumulations of certain metal ions together with the
simultaneous ion specific excretion rates from the body could lead to the
establishment of elevated concentrations of specific alloying elements of
implants. This could upset the overall balance established by physiological
tolerance to toxicity.
Two possible toxicity routes for metal ions released into body fluids due to corrosion and
wear.
The biological effects of certain metal ions that may be leached in vivo due to corrosion.