Introduction to Biomaterials

A biomaterial is a nonviable material intended to interface with a biological or physiological system in order
to evaluate, treat, augment, or replace any tissue, organ, or biological or physiological function
o Examples: pacemaker, prosthesis
o Donated organs are NOT biomaterials
o Example: eyeglasses vs. contact lens
Biomaterials science and engineering is the study of the relationship and integrated roles between
processing parameters, structure and composition, and properties on the performance of a material and a
biological or physiological system of a particular application.
o Example: parameter pyramid
Processing is the steps for handling and converting materials (and biological components) to form the
desired shape of the biomaterial
o Examples: patterning patch of adhesive protein on cell culture plates
o Criteria: how do the processing conditions (temperature, pH) affect the components?
Structure is the spatial arrangement of atoms within a material or biological or physiological system
o Examples: FCC or BCC, helix or sheets or random coil for proteins
Composition is the chemical makeup of a material or biological or physiological system
o Examples: atoms in biomaterial (titanium, gold), molecules in the biological component (amino acids)
Properties are the behavior or response of a material and/or biological or physiological system
o Examples: mechanical properties (yield point of a material), binding or adhesion strength of particular
proteins for particular cells
o Criteria: environment (temperature, pH, applied forces)
For bio-related applications of materials, one must consider how the viable components of the biological or
physiological environment respond to the presence of a foreign, nonviable (synthetic) material
For medical applications (implanted, injected), one must be concerned with in vivo (in the living) responses
o Example: inflammatory responseimmunological responsereject of biomaterial could occur or
infection could occur
For biotechnology applications (cell culture dishes), one must be concerned with in vitro (outside the living
body in an artificial environment) responses to the material
Important broad considerations for biomaterials:
1. Cytotoxicity involves the release of chemicals from a material that kills cells directly or indirectly
2. Biocompatibility is the ability or capacity of a material to perform with an appropriate host
response in a particular application
3. Healing (for implanted biomaterials) invokes a short term inflammatory response as particular
leukocytes (WBCs) are recruited to the injury site
4. Ethics minimize the risk to patients and considerations for animal testing; thus, the development of
biomaterials involves the following testing stages:
a. In vitro
b. In vivo tests in healthy animals
c. In vivo tests in model animals of particular disease or impaired state
d. Clinical trials (various stages)
5. Regulation is the time and cost considerations for implementing novel biomaterials which meet test
standards set forth by the following agencies:
a. FDA (Food and Drug Administration) provides approval for a specific device, not for a
general biomaterial
i. Example: contact lens vs. glasses
b. ASTM (American Society for Testing of Materials)
c. ISO (International Organization of Standardization)
Traditional approach (before 1970s) was biomaterials were designed to be inert with respect to the
physiological surroundings
Modern approach is to rationally design biomaterial to interface in a favorable, even dynamic, way with the
host
Classification of Materials Handout
Organelles Handout
Nucleic Acids Handout
DNA (helix) depends on histones to pack efficiently (complementary bases); whereas, RNA (single strand)
have self-complementary strands
After Binding to Biomaterial
What happens to cells after binding or attaching to the biomaterial via nonspecific or specific forces?
1. May affect general housekeeping activities of a cell
2. May affect cell viability (ability to live)
a. As cytotoxicity of biomaterial increases, cell viability decreases
b. If viability of a cell is compromised to a great extent, the cell may die
c. 2 routes to cell death:
i. Apoptosis: programmed cell death driven by the nucleus of the cell as part of the
bodys attempt to maintain homeostasis (normal status of the host); typically
involves the binding of specific ligands that signal a cell to die
1. The preferred route because there are no consequences with respect to inflammatory response
ii. Necrosis: cell death marked by significant changes to the cell structure and cell
membrane permeability
1. Cell often undergoes cell lysis (cell ruptures and releases its contents)
3. May affect cell stability
a. Labile cells can replicate or proliferate on a regular basis
b. Permanent cells are highly specialized cells that do not readily undergo replication (mitosis)
i. Examples: neurons, heart cells
c. Stable cells are somewhat specialized, but can be cued to undergo mitosis under particular
circumstances
4. May affect cell specialization
a. These specialized cells may be part of a particular tissue, organ, etc.
b. Cellular differentiation is the series of changes in a cell (normally involving gene expression)
by which a cell becomes specialized
i. Progenitor cells are nondifferentiated cells that are able to renew
ii. Totipotent stem cells can differentiate into any type of cell or cell phenotype
iii. Pluripotent stem cells can differentiate into many, but not all, cell phenotypes
iv. Embryonic stem cells are usually totipotent
v. Adult stem cells are usually pluripotent
1. Hematopoietic stem cells are derived from the bone marrow; form blood cells
2. Mesenchymal stem cells originate and differentiate into cells as part of connective tissue (tendons,
ligaments)
c. Stem cellscellular differentiationcommitted cells (specialized)
d. All cells in a host have the same genetic code (genome), but as cells differentiate, some
subset of genes are accessed in order to allow some subset of genes to be preferentially
expressed, while others are turned off
i. Results in distinctive behavior (biological profile) of a cell to give rise a particular cell
phenotype
ii. A cell phenotype can be determined by features such as the cell shape (cell
morphology)
iii. However, cell phenotypes can always be determined by its gene expression profile
5. Adherent cells (cells that differentiate and attach to a biomaterial) may organize themselves into
tissue
a. Requires a source of nutrients and waste disposal, particularly if multiple layers of cells are
involved
b. Angiogenesis is the formation of small blood vessels
6. Biomaterial in contact with cells, or even just near cells, may cause a distinctive reaction by the host
to this foreign material or invader which may involve
a. An inflammatory response which is an attempt by the host to dispose of foreign invaders
b. Fibrous capsule formation: cells of the host attempt to isolate the foreign invader by forming
a protective barrier-ultimate resolution
Specific vs. Nonspecific Forces or Interactions (Energy)
Nonspecific Forces
o Ubiquitous and does not involve recognition between particular functionalities or chemical groups
o Origins: primary bonds (covalent, ionic/coulombic or electrostatic, metallic), secondary bonds
(hydrogen, van der waals), and other bonds (hydrophobic, hydrophilic, steric)
o Nature:
Can be attractive or repulsive
Can be large or small in magnitude
Can be long-range or short-range in terms of distance dependence
o DLVO Model:
Involves van der waals and electrostatic forces
Accounts for geometry size and composition of macrobodies but not the steric interactions
o Biomaterials are typically subject to nonspecific attractions to proteins : protein adsorption
Can involve undesirable subsequent interactions with leukocytesinflammationbiomaterial
rejection
To overcome nonspecific attractive interactions between protein and biomaterial, need to promote
nonspecific repulsive interactions between protein and protected biomaterial by adding source of
steric repulsion (PEO, PEG)
Specific Forces
o Particular to a system (biological or physiological)
o Involves recognition between 2 or more particular adhesive groups
In case of cells, these matching adhesive groups or functionalities can be referred to as a receptor-
ligand pair: typically macromolecules (proteins, glycoproteins)
Ligand: often soluble species, but can also be mobilized on a second cell
Receptor: often present on cell membrane typically immobilized, but can undergo lateral diffusion
Examples: folate receptor-folic acid, antibody-antigen
o Origins: lock and key analogy
1. Particular structure to form the binding pocket in receptor for matching ligand
2. Favorable orientation between binding pocket of receptor and matching domain of ligand
3. Close association or contact between receptor-ligand pair
4. Involves nonspecific forces, but localized to particular regions of binding pocket and domain of
ligand (induced fit)
o Nature:
Usually attractive
Can be large or small in magnitude
Only short-range
o Receptor-ligand model accounts for binding and unbinding events between soluble pairs, but not
collective receptor-ligand interactions between 2 cells
High k
A
favors binding and low k
A
represents weak affinity between the receptor and the ligand
3 routes for an implanted biomaterial
o Unmodified surface: nonspecific attractive interactions with proteins are likely
o Surface modification #1: add grafted PEO chains to surface
o Surface modification #2: to promote specific interactions with particular proteins or even cells
(phenotypes) by functionalizing biomaterials with particular ligands
Experimental Assays to Measure Adhesion Strength, Cytotoxicity, and Viability
Common methods to measure the adhesion strength between cell and biomaterial involve measuring the
critical shear stress necessary to detach 50% of adhering cells from a biomaterial (plot)
Methods to measure adhesion strength:
1. Spinning disk: cells at edge are most likely to fall off
2. Parallel plates in a flow chamber: cells are either on top of bottom plate or on bottom of top plate
a. Cells are exposed to a small, but non-zero, fluid velocity so that only weakly adherent cells with
detach
In vitro tests
o Migration assays track the time dependent movement of cells
Individual cells (cells might move in random pattern)
Translocation speed is the speed of a cell in one general direction before it changes direction
Persistence time is the time that cell spends moving in one general direction before drastically
changing direction
The translocation speed and persistence time can be affected by adding soluble factors in one
area of the sample chamber (adding chemical gradient)
Soluble factor will eventually diffuse within fluid
Adding chemical gradient promotes chemotaxis: preferential path due to soluble factors
Groups of cells
Capillary tube test (Figure 9.41)
Boyden chamber assay (Figure 9.43)
o Cell morphology assays
If A has smaller spread area than B, B cells are more viable
Adhesion of cells is lower in A than B
Nonspecific: if plasma membrane is a negatively coated surface with positive charge, make plasma
hydrophobic
Specific: add ligand
Cytotoxicity assays are important in determining the overall biocompatibility of biomaterial
o Often arise from chemical factors from biomaterial (ions, molecules that have leached out of biomaterial;
leftover monomer or other reactants from biomaterial; corrosion by-products)
o Other factors include surface topography, biomaterial size and shape
o Many cytotoxicity assays are qualitative in nature (monitor changes in cell shape), but many are
quantitative in terms of measuring cell viability
o For quantitative assays
Use a dye that selectively labels only the dead cells OR use vital stain to only label live cells
Necrosis-affected cells are more likely to allow this
Measure intracellular enzyme that is now present in extracellular space due to cell death
Necrosis more likely to allow this as well
Example: lactate dehydrogenase (LDH)
o 3 types of in vitro cytotoxicity assays for measuring cell viability
1. Direct contact assay places the biomaterial in direct contact with the cells and divide into separate
ones
a. Sample chamberplace cells on chamber wallplace biomaterial on top of cells
b. As time passes, see how cells have migrated by splitting plate into circular zones to measure
spread area of cells
c. Placing biomaterial may crush cells (many cells in middle zone usually die from trauma instead of
cytotoxicity effects)
2. Agar diffusion assay
a. Agar is soft hydrogel that allows permeations and acts as a physical barrier and diffusion barrier
b. Cells are no longer subject to the trauma from direct contact, but are exposed to soluble factors
escaping from biomaterial
3. Elution or extract assays
a. Put biomaterial in solvent
b. After certain time interval when products from biomaterial has leached into solvent, remove the
biomaterial
c. Pour supernatant into a chamber with cells present
Introduction to Proteins
Proteins are polymerized amino acids (amino acid~monomer unitcondensation polymerization (lose
water)polymer chains (peptide, polypeptide, or protein)
Peptide: short chain of amino acid residues (typically too short to possess a 3D structure)
Polypeptide: longer chain of amino acid residues (typically has a defined length and sequence)
Proteins: usually refers to natural polypeptides of significant length and weight (have a defined 3D
structure under physiological conditions that helps promote its particular function)
Amino acids: building blocks for proteins (usually find 20 different types of amino acids in nature)
o Structure: N-terminus is the basic group (accepts hydrogen ions), and C-terminus is the acidic group
(donates hydrogen ions)
o Amphoteric: possess both cationic and anionic charges
o R groups can be 1 of 20 possibilities that fall into 1 of 4 types of amino acids
Nonpolar (R=CH
3
, XCH
3
)
Polar (R=XOH, XSH, XCONH
2
)
Acidic (R=XCOOH or XCOO
-
)
Basic (R=XNH
2
or XNH
3
+
)
o Polymerization of amino acids occurs via condensation reaction to form a peptide bond
o With 20 amino acids, many combinations of sequence composition and length depending on solution
conditions give rise to protein function
Protein structure
o Primary structure depends on linear order of amino acid residues in a chain
For natural proteins, primary structure is dictated by codons in genome
Codons are sets of 3 oligonucleotide bases which code for a particular amino acid
Recall: protein synthesis steps
DNA sequence information is encoded in mRNA via transcription
mRNA is used to synthesize a protein via translation
o Secondary structure arises from interactions between neighboring amino acid residues
Random coil has no regularity in structure
helix: 3.6 amino acid residues per turn; single-stranded; R groups face the outside of the helix
with hydrogen bonding occurring between every 4
th
R group; hydrogen bonding is not very strong,
allowing flexibility
Example: myosin proteins in muscles have helices
sheets allow for lamellar chain folding; stabilized by hydrogen bonding between folded domains;
found in semicrystalline polymers
Parallel vs. anti-parallel
R groups tend to be small to minimize steric hindrance to sheet formation
Hydrogen bonds are very important
Extended chains resist further extension
o Example: fibrin protein which is part of silk
o Tertiary structure: 3D arrangement of a polypeptide chain due to folding
Folding exposes particular amino acid residues to surrounding solve, hides particular amino acid
residues, and/or ultimately determines the formation of binding pockets for receptor-ligand to bind
to
Gives rise to particular functions of proteins
How protein folds into particular tertiary structure depends on primary and secondary structures as
well as the types of bonds and interactions on neighboring amino acid residues
Bond types
Primary: ionic bonds between acidic and basic groups and covalent bonds between cysteine
groups forming disulfide bonds
Secondary: hydrogen bonds and van der waals
Other types: hydrophilic (molecules that prefer to be solvated by water molecules) and
hydrophobic (molecules that tend to exclude water molecules)
Note that folded structures are dynamic and depend on solution conditions as well as its bound
states (induced fit that occurs in receptor-ligand binding events)
Tertiary structure can be disrupted (denatured) if the surrounding conditions change or if you
introduce a biomaterials surface
Denaturation of proteins is typically irreversible, but renaturation is possible for oligonucleotides
In its natural (native) state, many proteins have the simplest tertiary structure known as globular
(essentially spherical in shape)
o Quaternary structure results from having multiple polypeptide segments (subunits), each with their
own primary, secondary, and tertiary structures
Example: Hemoglobin is an oxygen-binding protein on RBCs
The primary structure of normal vs. sickle cell hemoglobin protein differs by only one amino acid
residue out of 149 amino acids (valine instead of glutamic acid)
Causes RBC to have a sickle cell shape that interferes with its function
Protein absorption is important to biomaterials because it is likely to alter the structure and function of the
protein, which could alter the biomaterial
Surface that resist, hinder, or at least delay protein adsorption events from occurring are known as
nonfouling surfaces (or stealth surfaces)
Nonfouling surfaces are often desirable because they prevent the subsequence recruitment of inflammatory
cells, but they also prevent the recruitment of desired cell phenotypes
Thermodynamic Factors for Protein Adsorption Events
Adsorption refers to the adherence or binding of a species to a surface
Absorption refers to the uptake of a species into the bulk of a material
G
ads
=H-TS
o G
ads
represents the change in Gibbs free energy due to a change in state from nonadsorbed state to
adsorbed state
o H represents the change in enthalpy (bond formation greatly affects enthalpy or internal energy of
the system)
o T is the absolute temperature
o S is the change in the entropy
As G
ads
becomes more negative, the event becomes more favorable
Favoring adsorption events
o H is negative: arises from bond formation between protein and surface (ionic bonds and van der waals)
o S is positive: water deadsorption or dehydration
Opposing, preventing, or hindering adsorption events
o H is positive: resistance to release bound water molecules (hydrophilic surface)
o S is negative: steric repulsion at the surface due to adsorbed or grafted polymer chains
Common synthetic polymer that is used in nonfouling surfaces is PEG
Though nonfouling surfaces are often desirable, immobilization of a functional protein to a surface is often
necessary for many biomaterials applications or biosensors and characterization techniques (ELISA)
One approach to immobilize a protein in a controlled manner involves using amino acid residues not directly
involved in a particular function
o Native protein with hydrophilic and hydrophobic domains as well as a binding pocket for specific
ligandallow for uncontrolled protein adsorption to biomaterial surfacedenatured protein does not
preserve desired function (hydrophobic domains bind to hydrophobic biomaterial surface)
o Protein with cysteine groupscontrolled immobilizationthiol groups on cysteine residues have high
affinity for gold surface
Kinetics of Proteins
Key considerations
1. Transport mechanism for protein to arrive at surface
a. Diffusion is the random movement of atoms, molecules, macromolecules, etc. through its
surroundings (fluid, tissue)
i. Macrobody diffusion described using Stokes-Einstein equation

ii. D=diffusion coefficient (diffusivity), k=Boltzmanns constant, T=absolute temperature, r=radius of
macrobody, =viscosity of fluid
iii. Can describe Brownian motion
b. Thermal convection: transfer of potential energy (heat) via currents within a fluid phase
c. Flow is the continual movement of a fluid medium under stress (blood flow through blood vessles)
i. Diffusion and flow are the primary transport mechanisms
d. Coupled transport typically involves 2 individual transport mechanisms (diffusion+flow)
i.

)
ii. v=velocity, Q=volumetric flow rate, b=distance between plates, w=width of plates, y=distance of
reference object from 1 wall
iii. relies on flow initially, then relies on diffusion to get to the wall
2. Mass or concentration of protein
3. Affinity (specific or nonspecific) of protein for surface
4. Presence of other competitive protein species
a. For 2-4 collectively, if protein A has low affinity for surface and is present in solution at high
concentration, and if protein B has high affinity for surface and is present in solution at low
concentration
i. Protein A will begin to occupy available sites (adsorption)
ii. Adsorption rate will decrease as fewer sites are available
iii. Surface is saturated with protein A
iv. Competitive displacement of protein A by protein B
b. Know the plots
c. Replacement of weaker adsorbate by a strong adsorbate species over time is called the Vroman
effect
5. Conformational and biological changes in adsorbate (species that has adsorbed to surface)
a. Change in protein structure often involves denaturation of protein
b. Nonspecific affinity of protein for biomaterial tends to increase as
i. Size of protein increases
ii. Net opposite charge between protein and surface increases
iii. Lack of net charge on protein and/or biomaterial
iv. Protein structure becomes more static upon adsorption to surface
Separation and Identification of Proteins in a Mixture
Basis of protein separation from a mixture
o Size (and/or molecular weight)
o Affinity for a substrate
o Charge
Separation based on size alone
o Size exclusion chromatography (SEC)
Proteins migrate through a column of porous resin beads using gravity (and/or pressure) as the force
Larger species take less time (only needs to go between beads)
Smaller species take more time (have to go through and between the beads)
Solvent is the mobile phase and beads is the stationary phase
o Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
SDS is a surfactant oligomer used to make intrinsic net charge to provide an equivalent charge:mass
ratio for all proteins
Protein denatures into more linear conformation
Each SDS oligomer binds to 2 amino acid residues in a regular fashion
Proteins migrate through a continuous hydrogel (highly entangled, cross-linked polyacrylamide
chains)
Smaller ones elute faster
Separation based on affinity
o Specific binding to immobilized ligand on resin bead
o Nonspecific affinity based on hydrophobic attractions between hydrophobic bead and hydrophobic
domains of protein of interest
Separation based on charge
o Ion-exchange chromatography
Identification of protein of interest
o Chromatography is based on color changes due to photon absorption characteristics of
macromolecules, often at a particular wavelength of light
Employs chromophores which are sensitive (chemically) to light
Colorless chromophore prior to light exposureadd enzyme to drive a light-sensitive
reactioncolor change due to photon absorption
o Fluorescence is based on photon emission characteristic of macromolecules or dye molecules attached
to macromolecule of interest
Fluorophore is usually molecule dye that reaches an excited electronic state through an excitation
event, then emits photon upon relaxation to lower energy state or ground state
o Western blotting
Separate via SDS-PAGE
Transfer the bands of separated protein species onto nitrocellulose paper
Fluorescence tagging of protein of interest
Wash off any unbound ligand
o ELISA (Enzyme-Linked Immunosorbent Assay)
Separate via SDS-PAGE or affinity chromatography
Plate well for introducing a solution of protein
Use enzyme-linked chromophore on ligand to tag the protein of interest
Wash off any unbound ligand
Add light to catalyze reaction
o Sandwich ELISA
Add ligand #1 to the wells
Add separated protein and wash off any unbound protein
Add ligand #2 that has enzyme-linked chromophore
Wash to remove any unbound ligand #2
Add light to catalyze reaction
Separation and Identification of Oligonucleotides in a Mixture
Southern blotting: often used to identify sequence coding for a particular gene
o Denature DNA duplexes and add base (NaOH)
o Electrophoretic separation (PAGE)
No SDS is needed because already equal mass:charge ratio
o Identify the sequence of interest using fluorescently-tagged complementary sequence
o Wash away any unbound sequences
Northern blotting: separate and identify RNA sequence
o Electrophoretic separation
o Identify using fluorescently-tagged complementary sequence
Amplification of oligonucleotides
o Polypeptide synthesis is complicated, but if you have at least one copy of an oligonucleotide sequence,
you can copy or amplify the number of sequence strands using polymerase chain reaction for DNA or
reverse-transcription PCR for RNA
o Steps for PCR
DNA duplexdenature via heatadd primers (short complementary sequences) and monomers of
A,T,G, and C (nucleotides)cool solution to allow for hybridizationadd polymerase to form
phosphodiester bonds between nucleotidesduplicated duplex
o Amplify the number of copies of a duplex by 2
x
where x is the number of cycles
Overview of Hosts Defense System
The immune system is triggered by injury to defend the host against infections, organisms, and foreign
material and to maintain or return to homeostasis
Self (host-derived) vs. non-self (foreign)
Foreign material could be (virus, bacteria), biomaterial, host cells infected by foreign pathogen, etc.
Extent of injury depends on chemical nature, size, shape, surface roughness, and other physical properties of
biomaterial
Extent of immune response depends on the nature and extent of injury
Types of Immune Responses
Innate/intrinsic immunity
o Born with it
o Nonspecific in nature: host identifies a species as non-self (foreign), but does not recognize specificity
of the foreign pathogen
o 1
st
line of defense
Adaptive/acquired immunity
o Generate a memory for past foreign invaders
o Specific in nature: body prepares a particular set of cells (lymphocytes) that recognize particular
elements of particular foreign pathogens
o Invoked if the innate response is unsuccessful in clearing the host of a foreign material
Other general differences between innate and adaptive immunity
o Physical barrier
Innate: skin, mucosal lining (nose)
Adaptive: lymphocytes in epithelia
o Chemical and other physiological barriers
Innate: body temperature (fever), antimicrobial proteins, low pH in stomach
Adaptive: antibodies (secreted by B cells)
o Other chemical barriers
Both have cytokines: cellular proteins (usually secreted) which affect the behavior of other cells
Examples: interleukins (IL)-cytokines secreted by leukocytes (WBCs)
Lymphocytes are a subset of leukocytes
Chemokines: chemoattractant proteins that control the adhesion, chemotaxis, and activation of
leukocytes/lymphocytes
o Types of cells involved
Innate: leukocytes, neutrophils, eosinophils, basophils
Adaptive: lymphocytes (T cells for cellular immunity and B cells for humoral immunity)
o Cells that can act as intermediaries between innate and adaptive immunity
Innate: macrophages internalize pathogens via phagocytosis and digests the pathogens
Adaptive: macrophages present digested foreign antigen on plasma membrane to signal specific
lymphocytes (aka antigen-presenting cells)
Types of Leukocytes
Granulocytes or polymorphonuclear (PMN) leukocytes
o Granules in the cytoplasm identified via staining
o Irregularly shaped multiple nuclei
o Main function is to phagocytose foreign material as well as lysed cell parts
Example: neutrophils, basophils, and eosinophils
Neutrophils die very quickly following their activation and phagocytic activity
Macrophages also phagocytose foreign material, but they do not die quickly in case they need to act
as APC
Monocytes
o One nucleus
o Circulate throughout bloodstream and usually only differentiate when recruited to infected, inflamed
area or as they enter tissue
o Often differentiate into macrophages
o Phagocytic capabilities, but macrophages have more capabilities
Lymphocytes (particular to the adaptive immune system)
o T cells originate in thymusactivationeffector T cells used in cellular immunity
o B cells originate in bone marrowactivationeffector B cells (plasma cells) secrete antibodies
Megakaryocytes
o Originate in bone marrow
o If activated, they break up into smaller fragments called platelets along clotting cascade
Overview of Inflammation
Typically the first defensive response of a host to a biomaterial always involves an inflammatory system
Acute inflammation is an immediate response of short duration (hours, days); typically a normal response
that persists for days to years
o General undesirable because no resolution of the injury is reached via chronic inflammation
o Cardinal signs
1. Tumor (swelling) is caused by recruitment of inflammatory cells (neutrophils, macrophages)
2. Dolor (pain) is caused by kinins release during injury (blood clotting cascade)
3. Calore (heating) is due to increased blood volume; part of the bodys attempt to kill foreign
pathogens near injury
4. Rubor (redness) is due to increased blood volume near injury site
o Steps
Activated tissue macrophages secret cytokines (IL-1, IL-6, TNF-) which increase vascular
expressions of CAMs to recruit neutrophils and other inflammatory cells
IL is interleukin and TNF is tumor necrosis factor
Nearby blood vessels vasodilate or expand in diameter to allow blood volume to increase and thus,
faster recruitment of inflammatory cells to injury site
Neutrophils are recruited and activated for phagocytosis (typically occurs within 1
st
few hours
following vasodilation)
Activated macrophages are recruited
Phagocytosis typically occurs 5-6 hours following injury
Conditional: IF the injury is resolved, terminate acute inflammation by decreasing inflammatory
stimulus (turn off inflammatory stimulus by introducing competing, but non-inflammatory
inducing cytokines such as IL-1ra)
ra stands for receptor agonist
competes with IL-1 for receptor on inflammatory cells
CAMs Handout
Selectins
o Glycoproteins which recognize specific carbohydrate groups called mucins
o Some are expressed on inflamed vascular endothelial cells (E-selectin, P-selectin)
o Some are expressed on inflammatory cells (neutrophils)
o Responsible for initial, weak adhesion events between leukocytes (neutrophils) and vascular
endothelial to drive neutrophil rolling along vasculature near injury
Mucins
o Ligand to selectins
Integrins
o Heterodimeric proteins present on leukocytes (neutrophils)
o Bind strongly to their ligand known as immunoglobulin (Ig) superfamily
o Expressed by inflamed endothelial cells near injury
ICAM-1, VCAM-1
Equilibrium constant
Important cytokines responsible for vascular changes during inflammatory response
o TNF-, IL-1, IL-6
Secreted by activated macrophages
Increase CAMs expression level on vasculature (endothelial cells)
Increase vascular permeability
Important chemokines responsible for activating inflammatory cells
o IL-8
Induces a conformational change in integrins to allow binding to its ligand (Ig superfamily)
Neutrophil Recruitment
1. Rolling of neutrophils along endothelial cells due to transient adhesion events from weak mucin-selectin
binding events
2. Arrest or stop neutrophil rolling via strong or tight binding events between integrin-Ig superfamily
receptor-ligand pairs
3. Diapedesis is the movement of neutrophils across vasculature by squeezing between leaky endothelial cell
junctions
a. Favored by increased vascular permeability
b. Favored by chemoattractants (subclass of chemokines) especially IL-8 and MIP-1b which cause
chemotaxis
4. Transendothelial migration is the process of movement across endothelial cell lining through surrounding
tissue to actual injury site
Extravasation is the entire series of steps involved in recruiting neutrophils from blood stream to injury site
Role of neutrophils
o Following their arrival at injury site
Phagocytosis of foreign material
Respiratory burst: release of reactive O
2
and N
2
series to kill foreign pathogen
Role of macrophages
o Residual tissue macrophages are already present
o Additional macrophages are recruited following neutrophil recruitment (source is the circulating
monocytes that differentiate into macrophages)
o Actual functions
Phagocytosis: macrophages have the highest capacity for phagocytosis of foreign materials
Release of cytokines to upregulate key CAMs on endothelial cells near injury that are necessary to
recruit other leukocytes
Conditional: IF injury is not resolved via acute inflammatory response, macrophages serve as APCs
to recruit lymphocytes as part of AIR
Normal Wound Healing
Acute inflammation and blood clotting cascade (simultaneous)
o Short duration
o Primarily involves neutrophils and macrophages
o Phagocytosis (engulfment) and degradation of foreign material (dependent on size of biomaterial)
o Frustrated phagocytosis by macrophages involves release of chemically degrading products in an
attempt to degrade or breakdown larger biomaterials
Formation of granulation tissue (2-5 days following injury)
o Macrophages and fibroblasts of vascular endothelial cells are recruited to the injury site to form
granulation tissue
o Granulation tissue has pink, soft appearance and is specialized tissue which is a hallmark of
inflammation that is healing
o Role of phenotypes
Macrophages secret chemoattractants to recruit fibroblasts and endothelial cells
Fibroblasts synthesize extracellular matrix proteins (proteoglycans, collagen) and later forms
fibrous capsule
Endothelial cells are responsible for angiogenesis by promoting building of new blood vessels from
existing ones
Foreign body reaction
o Involves granulation tissue and FBGCs
o Foreign body giant cells (FBGCs) are fused macrophages and monocytes
Can phagocytose larger objects
Typically around injury site for the lifetime of the implant
Fibrous encapsulation
o Comprised of mature granulation tissue consisting of larger blood vessels and aligned collagen fibers
o Marks end of healing stage
o Serves as barrier between biomaterial and host (physiological environment)
Factors Involved in Injury
Size of biomaterial
o As size increases, more frustrated phagocytosis (mostly macrophages) and more FBGCs form for more
phagocytic capabilities
Shape
Surface area
o More surface area means more FBGC formation
o Less surface area means less FBGC formation and more granulation tissue formation
Implant site
o Splinter in the skin (no blood, but painful)
If injury involves only epithelial cells, then extrusion may occur in which the skin forms a pouch
around the material to extract it out
o If the injury involves labile or stable cells, then either fibrous encapsulation or regeneration is possible
in which damaged cells are replaced by cells identical to the native tissue
o If injury involves permanent cells, repair is possible in which damaged cells are replaced by cells that
are different from original native tissue (scar tissue formation is likely)
Mechanical factors such as rubbing, shedding, wearing into smaller pieces
Chemical nature of biomaterial
o If material is biodegradable, material may completely disappear and undergo resorption
o If material is not degradable and recognized as foreign, fibrous encapsulation is likely
o If material is not degradable, but can completely interface with physiological surrounds, integration
may occur without fibrous encapsulation
o Fibrous encapsulation of implanted biomaterial is the most common resolution (new equilibrium state
or homeostasis redefined for host)
Repair vs. Regeneration
For superficial wound involving the epidermal layer, regeneration may occur
o Epithelial cells near wound degrade surround extracellular matrix and then flatten out to cover some of
the wound
o Migration and proliferation of epithelial cells to wound are to cover any remaining exposed areas
o Epithelial cells resume their normal morphology and now have to reattach to extracellular matrix
For deep wounds in the dermal layer, repair will occur
o Acute inflammation and blood clotting cascade which forms a barrier to stop bleeding and prevent
foreign pathogens from entering wound area
14 clotting factors and platelets form a polymerized fibrin network (scab)
o Formation of granulation tissue involving influx of fibroblasts
Deposit collage fibers (type III) in random orientation near extracellular matrix
o Dissolve fibrin clot via enzymatic cleavage events and scab falls off
o Scar formation due to constant remodeling of extracellular matrix
Randomly oriented collage type III fibers replaced by oriented collagen I fibers
Collagen accumulation and remodeling occurs for 2-3 months
Chemical and physical properties of biomaterial may induce a persistent inflammatory stimuli (chronic
inflammation)
o Undesirable because it doesnt resolve injury
o Can last weeks, years, to lifetime of host
o Macrophages, monocytes, lymphocytes, and granulomas (layer of FBGCs) involved
In Vivo Testing of Inflammatory Response
In vivo testing first occurs in live animal models
o Starts off with rats, mice, rabbits
o Later tests in sheep, dogs, cows, pigs
Important factors include choice of animal, implant site, and size of biomaterial
Response is examined at particular time intervals
o Acute toxicity effects are negative effects within 24 hours following administration/implantation
o Subacute toxicity effects are within 14-28 days
o Subchronic toxicity effects are within 90 days
o Chronic toxicity effects are after 90 days
Choice of administration of biomaterial
o Direct contact: intact biomaterial makers intimate contact with surrounding tissue
o Extracts: pieces of biomaterial are implanted
o Cage implant: biomaterial is housed in a stainless steel cage to examine the inflammatory response in
the absence of contact between biomaterial and surrounding tissue
Assess in vivo inflammatory response via histology (taking tissue sections to examine cell phenotypes
present, tissue present, etc.)
Hallmarks of Acquired/Adaptive Immune Response
Involves specific recognition of particular foreign antigen (usually involve peptide fragments from foreign
pathogen)
o Each nave (not yet exposed but has capabilities) T cell and each nave B cell can recognize a specific
foreign antigen or unique antigen
o In contrast, innate immunity only recognizes general molecular patterns of foreign pathogen
Diversity in lymphocyte populations
o Due to genetic rearrangement during development and maturation of nave lymphocytes
o Large and diverse lymphocyte population that results
o If AIR is needed, APCs must find the particular T and/or B cell that recognizes the antigen present
Distinction between self and non-self: nave T and B cells that recognize self-antigens are quickly destroyed
during a selection process
Immunological memory
o Nave T/B cellsactivationeffector T/B cells and memory T/B cellsmitosisclonal
populationsuccessful cellular/humor IRregulation process to eliminate many effector cells, but
leave memory cells alone in case same pathogen invades againmemory cells outlive effector cells
Antigen: antibody generator (Ag)
Antibody: immunoglobulin type of protein (Ab)
o Secreted by plasma cells and immobilized on B cells
o Ab binds to a particular binding domain of a Ag known as an epitope
Types of AIR
o Humoral immune response
Plasma cells
Soluble antibodies secreted by plasma cells
Basic structure of typical bivalent (2 binding sites): heavy chains, light chains, disulfide bonds,
hinge region, antigen-binding fragment, constant fragment
Antigen-binding fragment binds to antigen to prevent antigen from its intended activity
Constant fragment binds to particular receptors (FCR) on macrophages to alow phagocytosis of
Ag-Ab complexes
Role of secreted antibodies
Antibody binding to antigen to inactivate it and prevent it from infecting a host cell
Antibody-antigen complex is marked for phagocytosis by macrophages
Antibody binding to antigen can invoke complement system: set of proteins can self-assemble
on pathogen surface to puncture a hole
o Cellular immune response
T cells can split into helper T cells and cytotoxic T cells
Cytotoxic T cells (CD8 receptor) recognize and kill non-self or otherwise altered host cells
Helper T cells can either help to mediate immune response of plasma cells or help cytotoxic cells
(CD4 receptor)
Survey of Homeostasis
Homeostasis: normal physiological activity in the absence of injury to retain the healthy status of the host
Hemostasis: mechanisms by host to arrest or stop bleeding which involves the formation and ultimate
dissolution or degradation of blood clot
Thrombosis refers to the formation of a blood clot (thrombus)
o In the absence of biomaterials, thrombus can form in blood vessels
o In presence of biomaterials, thrombus can form on the biomaterial itself
o Local/systemic effects of thrombosis according to where it forms
Hemostasis overview
o Injury occurs: damage to blood vessels and surrounding tissue
o Platelets migrate to injury site (activated by binding to exposed collagen)
o Activated platelets release granular contents containing chemotactic factors that activate and recruit
other platelets to injury site as well as other molecules (fibrinogen monomer)
o Fibrinogen polymerizes to form fibrin network surrounding platelets via thrombin and Factor XIIIa
(a=activated)
Soluble fibrinogen is the monomer
Thrombin (secreted by activated platelets) cleaves fibrinogen at key sites to expose polymerization
sites to form fibrin monomer
Exposed monomers now form hydrogen bonds with one another to form long fibers that assemble
and branch
Factor XIIIa and calcium ions promote the formation of strong covalent bonds as fibers cross-link
and for the fibrin polymer (aka fibrin network)
To prevent blood clot formation, add chelating agens (heparin) to form a complex with calcium
ions
o Forms stable platelet plug which fills the wound area and serves as a scaffold for later repair
Scaffold is only temporary; ultimately, the fibrin network is enzymatically broken down during
fibrinolysis via plasmin
Hemostasis in presence of biomaterial (in contact): platelets often stick to biomaterial and form thrombus
by recruiting other platelets and molecules for fibrin network
Platelets
o Little plates 3-4 micrometers in diameter
o Non-nucleated fragments of megakaryocytes
o Functions
Initially stop or arrest bleeding by adhering to biomaterial surface (if relevant) and/or exposed
collagen: platelet adhesion
key mediators of adhesion to collagen
o receptors on platelets: GP1b
o von willebrand factor (VWF) on collagen
conformational changes to integrin receptors on now activated platelets
further strengthens adhesion between platelets and collagen
promote adhesion to other platelets
induce platelet-platelet aggregation
Hybrid Orbitals
Valence shell: the last orbital/shell in an atom to be filled by electrons or the first orbital/shell to lose
electrons
o Valence electrons that occupy valence shell are most likely candidates for participating in bonds and
excitation events
In several cases where atoms bond to other atoms, the valence shell itself is not a single orbital, but rather a
hybrid orbital
o Sigma and pi bonds
o sp, sp
2
, sp
3

o Pi bonds prevent bond rotation, but are easy to break and excite to a higher energy state
Overview of Fluorescence and Absorption
Valence electrons participate in bonding (primary bonds) and in absorption/excitation events
Pi bonds are of particular interest for bond formation and excitation
Fluorescence
o Molecular dyeelectronic states
o Excitation light source at particular excitation wavelength causes electron to jump from ground state to
an excited electronic state
o Nonradioactive decay occurs involving relaxation of electron to a lower, excited electronic state
o Fluorescence event occurs in which electron relaxes back down to ground state and releases a photon
emission
o For quantum dots comprised of semiconductor materials, typically as size increase, the energy gap
decreases (E~1/)
Absorption
o Electron is excited from pi state to pi antibonding state
o UV-VIS compares relative absorption as a function of wavelength
Useful for SEC
Useful for determining concentration of macromolecules
Peak increases as concentration increases
Short-Range and Long-Range Order in Solids
All solids (crystalline or amorphous/noncrystalline) exhibit SRO in which nearest neighbor atoms have a
specific spatial arrangement
Coordination number is the number ratio of one reference atom t all of its nearest neighboring atoms
(relevant for both SRO and LRO)
For crystalline materials with LRO, we can assume
o Periodic arrangement or regular spatial placement of atoms in crystal, crystalline lattice, or lattice
o A lattice point refers to an individual point (typically one atom) within a crystalline lattice
Each lattice point is surrounded by an identical arrangement of neighboring points
Typically refers to a single atom whereas a basis refers to a group of atoms associated with a lattice
point
o Unit cell is the smallest group of lattice points that completely describe the structural arrangement of a
crystalline lattice
Lattice parameters used to describe the structural arrangement of lattice points in a crystal
o Edge length is the length of an edge in a unit cell
o Angle between edges
Simple cubic, body centered cubic, face centered cubic
Close-packed direction: any direction in a unit cell that goes through the centers of touching atoms
o Close-packed families
FCC: {110}
BCC: {111}
SC: {100}
Close-packed planes: 2D cut/plane through the unit cell that includes the centers of atoms, each of which is
touching all of its nearest neighboring atoms
Close-packed direction and planes are important for calculating atomic packing factor and density of
material
Volumetric contribution for center is 1, for corner is 1/8, for face-centered is
CsCl crystal structure
o Not a unit cell
o Not electrically neutral
o Inner penetrating system of anion and cation
NaCl crystal structure
o Inner penetrating FCC
Defects Handout
Overview of Polymers (Handout)
Polymer or macromolecule is a chain of molecules linked together by covalent bonds via polymerization
o Can form new covalent bonds between different functional groups on monomers to form polymers
Water is likely to be a byproduct
Additional function groups on monomers allow the formation of branched polymers
o Can break pi bonds in unsaturated (has available pi bonds) monomers to form new covalent bonds in
polymer
If degree of polymerization, n, is small (2-10), then it is an oligomer (surfactant)
If n is large (>>10), then it is a polymer
o In reality, typically forms many chains with different values for n
Be able to draw structures in different notations
Conformation vs. Configuration Handout
Conformation: the overall shape and structural attributes that can be changed via bond rotation
Configuration: structural attributes for polymer that can be changed by breaking bonds
o For example, breaking bonds to rearrange R groups to change tacticity
General structure (polymer architecture)
o Linear chain: chain with 2 ends
Typically can melt from solid to liquid state
Chain flexibility, conformation, and crystallinity depend on
Concentration of polymer
o Dilute concentrations: many intrachain interactions possible within same chain
o High concentrations (heat polymer or polymer melt): both intrachain and interchain
interactions and bonds form
Chemical composition
o Steric effects arise depending on the size and placement of side groups
o Look at rest of handout
Formation of Polymer Networks
Utility of heat polymer networks (PMMA) and polymer networks that swell in water (hydrogels)
Be able to show electron movement in formation of a linear chain of PAAM with 3 repeat units
Be able to show how to cross-link polymer chains to form covalent bonds between chains
Copolymer
Up to now, we have discussed polymers comprised of only 1 type of repeat unit to form a homopolymer
If more than 1 type of monomer is used during polymerization: copolymer
Types of copolymers
o Random copolymer: AABBBBAABA
o Alternating copolymer: ABABABAB
o Block copolymer: AAAABBBBB
Self-assembly occurs in presence of water
If A was hydrophilic and B was hydrophobic
Short chain of A, long chain of B can lead to micelle formation
Long chain of A, long chain of B can lead to vesicle formation (commonly used in drug delivery)
Step vs. Chain Polymerization
Step polymerization
o Aka condensation polymerization
o Monomers have 2 or more reactive sites
o Polymer chains grow step-wise by reactions between ANY 2 reactive species (either monomer and/or
reactive chain)
o Monomer is used up quickly
o Byproduct often formed (water, HCl)
o Reaction continues (chain keeps growing) until no reactive groups are left
Chain polymerization
o Aka addition polymerization
o Monomers usually have the form CH
2
=CX
1
X
2

o Polymer chain grows ONLY by the reaction between a monomer and a growing chain
o Break unsaturated pi bonds to form new covalent bonds along polymer backbone
o Usually no byproducts form
o Involves distinct stages
Initiation
Creates free radical center initiator species which often has peroxide (-O-O-) or azo (-N=N-)
linkage
Propagation/growth
Typically head to tail addition, but can have head to head addition (sterically hindered)
Termination
Combination usually results in head to head addition of 2 active chains
Disproportionation occurs when one hydrogen atom is extracted from one growing chain by
another growing chain
Know how to draw polymer products for step and chain polymerization
Know how to show electron transfer in chain polymerization
Some polymerization routes have aspects of both step and chain polymerization
o Polyurethanes resemble step polymerization but not byproduct
Used in artificial organs
o One limitation for synthetic polymers is the cytotoxicity of monomers: cant polymerize in presence of
cells for cell encapsulation purposes
Need to remove any residual monomers
Avoid using cytotoxic synthetic monomer (use natural polymer such as alginate or gelatin)
Ultimately, its very likely that polymer batch itself will have chains with different molecular weights,
resulting from the inability to start and end polymerization at the same time throughout the reaction vessel
For a given chain i, the MWi or Mi=n*MWru (Problem Set)
o Where n is the degree of polymerization, MWru is the molecular weight of the repeat unit, and MWi is
the molecular weight of the chain
For a polymer chain with many chains, the average molecular weight can be describe with
o Number average molecular weight, Mn
Mn=(NiMi)/Ni
Ni is the total number of chains
Mn treats all chains equally
o Weight average molecular weight, Mw
Mw=Ni(Mi)
2
/NiMi
Mw permits larger/heavier chains to make a proportionally larger contribution to average molecular
weight value
In a polydisperse polymer chain in which there is a range of molecular weights, Mw>Mn
We indicate the range of molecular weight values in a batch using polydispersity index, P
I
=Mw/Mn
o For a monodisperse polymer batch in which all chains have the same molecular weight, P
I
=1
Diffusion In and Out of Biomaterials
Diffusion is the movement of atoms or other species through random jumping events
In a solid, diffusion requires
o Free space to jump into (in crystalline materials: vacancies, interstitial, voids)
o Sufficient thermal energy to overcome activation barrier to jumping

) or

o D is the diffusivity (diffusion coefficient), D
0
is the material constant independent of temperature, Q is
the activation energy barrier, R is the gas constant, T is the temperature
o Slope is Q/RT and the intercept is lnD
0

Though diffusion is a random process, over time, a species will diffuse over same finite distance
We can describe the net movement of a group of identical species across an interface to describe a
phenomena known as flux
o Flux is the net number of species that move across an interface or plane of a given unit area per unit
time
o Ficks 1
st
law

J is the flux, D is the diffusivity, and concentration gradient
Assuming that concentration gradient doesnt change with time
Many biomaterials that deliver therapeutic agents rely on diffusion alone for the therapeutic agent to leave
the material matrix
o Plot
Crystallinity in Polymers
Considerations
o Steric effects of side groups: increased bulkiness decreases percent crystallinity
o Increased chain branching decreases percent crystallinity
o Tacticity
o Copolymer: regularity of A and B repeat units
Chain folded model to describe semi-crystalline polymers
o Lamella is the crystalline domain, amorphous is the noncrystalline domain
o Spherulites are 3D aggregates of lamellae that radiate from a center
To create new dislocations or move existing locations, must induce plastic deformation
o In metals, to decrease the concentration of dislocations, anneal
o In polymers, to increase percent crystallinity, anneal
o Anneal: heat (provide energy for defects to move and/or for chains to become more mobile) to an
elevated temperature, not to melt the material
o Slowly cool to allow time for ordering of atoms, chains, etc.
If one quickly cools (quenches) the sample, it does not allow adequate time for ordering to occur in
material
Decreases percent crystallinity, especially in polymers
Temperature-dependent Phase Transitions
Purely crystalline metals possess clear phase transitions
o From solid crystal to liquid, melting temperature marks transition
Pure liquid phase
o Viscous response to applied stress
o Time-dependent deformation response to applied stress
o
is the shear stress, is the viscosity (resistance to flow/deformation), is the shear strain rate
Pure solid phase
o Elastic (reversible) response to small applied stress
o Independent of time in terms of deformation (assuming that temperature is not elevated)
o
G is the shear elastic modulus
Viscoelastic material possesses both solid-like and liquid-like responses to applied stress
o Characteristic of many polymeric systems
o

G* is the complex shear modulus, G is the shear elastic modulus, G is the shear viscous or loss
modulus
Pure solid allows us to recover the original structure prior to deformation and recover energy expended into
deformation
All energy expended into deforming a liquid is lost
Possible phase transitions in polymers assuming heating a fresh batch of polymer with no heating history
(increase temperature, increase chain mobility)
o T<Tg: glassy phase, brittle material (G)
o Tg<T<Tc: rubbery phase, viscoelastic behavior, solid and liquid amorphous solid (G*)
o Tc<T<Tm: solid phase is crystalline, solid and liquid likely (G*)
o Tm<t: liquid, fluid-like repones (G)
o Assuming only elevation of temperature
Various classes of materials
o Crystalline metal (crystal<Tm<liquid)
o Amorphous polymer
Linear chains that are not crosslinked (glass<Tg<viscoelastic rubbery phase<Tm<liquid)
Crosslinked linear chain-not enough mobility for chains to fold (glass phase<Tg<viscoelastic rubbery
phase)
o Semicrystalline polymer (glassy phase<Tg<rubbery phase<Tc<semicrystalline<Tm<liquid)
o Ceramic glass (glassy phase<Tg<more liquid-like, but never liquid-like in behavior)
For a ceramic glass, these materials are often handled/processed into final shape by processing
Working point: more liquid-like deformation
Differential Scanning Calorimetry
Measure the head needed (heat flow) to change the temperature of a sample and a reference at a particular
heating (or cooling) rate
Reference has a known specific heat capacity (amount of heat needed to raise the temperature of the
material by 1C)
At a particular temperature value corresponding to a phase transition in the sample, if the phase transition is
endothermic (requires heat input), then more heat is needed to maintain the same temperature as the
reference
o Glassy phase<Tg<rubbery phase
o Solid phase<Tm<liquid phase
If the phase transition is exothermic (heat is released), then less heat flow is needed to maintain the same
temperature for your sample as the reference
o Noncrystalline solid<Tc<crystalline solid
DSC involves monitoring changes in the heat capacity of a sample as it undergoes temperature dependent
phase transitions
Plot
o All 3 likely to occur in a semicrystalline polymer (not crosslinked)
Lower average molecular weight means lower Tm
Lower percent crystallinity means lower Tm
Degradation (Handout)
Following exposure to a physiological environment, biomaterial can be subject to changes in its chemical
nature and physical dimensions (over time, may disappear altogether)
Chemical degradation involves a chemical process resulting in the cleavage of bonds
Biodegradation is a degradation process due to cues from physiological environment
Physical erosion involves a physical process which leads to changes in the overall shape,size, or mass of the
biomaterial
o Swelling, deformation, weight loss, wearing, structural disintegration
Physiological cues that promote chemical degradation
o Ions present in blood (154 mM NaCl)
o Low pH in particular regions of cell or body (lysosomes, phagosome, stomach)
o Due to enzymes which reduce the activation energy barrier for a chemical reaction, but ultimately
enzymes are not consumed
o Actions of inflammatory cells (release of reactive species by neutrophils)
Chemical degradation of metals is called corrosion which is electrochemical process in which metal ions are
generated from neutral metal atoms
o Oxidation-reduction
o Can result in leaching of metal ions from biomaterial into body
o Can ultimately also result in new chemical product, especially on biomaterial surface
If the chemical product resulting from corrosion is relatively chemically stable, then it may act as a
diffusion barrier to prevent further corrosion and act as a passivation layer
Passivation layer formation is often possible in ceramic materials (ionic solids)
Polymer degradation (Handout)
o Overall mechanisms
Swelling and dissolution of polymer material (both physical processes)
Swelling occurs in hydrogels in aqueous solvent; however, complete dissolution of hydrogel is
prevented due to chemical crosslinks
Dissolution has no crosslinks present; break secondary bonds by adding water which can
promote subsequent chemical degradation
Chain scission: covalent bonds along the backbone, between backbone and side groups, or between
backbone and side chains are broken
Average molecular weight of polymer chains decrease
Always requires solvent (usually water) in order to cleave bonds
Usually requires presence of reactive species in order to cleave bonds
2 routes
o Via hydrolysis (water must be present) depends on
Reactivity of groups along polymer backbone, crosslinks, side groups, etc. (presence of
carbonyl group in many reactive chemical groups make them susceptible to cleavage)
Amount of water present (locally around each chain)
Percent cross linking (less percent crosslinkage means greater degradation rates)
At T>Tg, likely to promote degradation as diffusion and permeability are more favored
Less percent crystallinity promotes degradation
Increase in surface area to volume ratio promotes degradation
o Via oxidation (Handout)
o Copolymers can allow for the inclusion of multiple features to promote biodegradation (PLGA)
PGA: poly(glycolic acid)
Hydrophilic (promotes degradation)
Crystalline
Used for 1
st
suture materials because it is totally resorbable, but it degrades quickly
PLA: poly(lactic acid)
Hydrophobic
Amorphous (not as well packed so promotes degradation)
Degrades, but slower than PGA
Deformation of materials under applied load
Mechanical loads
o Tensile
Lf>Li, Af<Ai
Engineering stress or true stress
Engineering strain or true strain
Real sample is dog-bone shaped
o Compressive (metals, ceramics, polymers)
Lf<Li, Af>Ai
o Bending (ceramics)
Maximum tensile stress at top middle and maximum compressive stress at bottom middle
Derivative of this bending test for biomaterials is called the knot test
o Shear (polymers, liquid-like properties such as suspension of solid particles in liquid)
Shear stress and strain
Recall shear elastic modulus
o Oscillatory
Tensile and compressive
Tensile and no load
Shear
o Torsion
Types of deformation that can result (assessed after the load is removed)
o Elastic deformation: material fully recovers its original structure and dimensions after load is removed
o Plastic deformation: material undergoes permanent change in structure as a result of applied load
Tensile testing is one of the more common mechanical tests for assessing deformation
o Plots of ideal metals, semi-crystalline polymers, and ceramics
Youngs modulus, E
Strain value range for elastic deformation
Strain value range for plastic deformation
Yield strain value (and relation to Youngs modulus, E-stiffness)
Measures of ductility or the ability to draw/elongate a material without fracturing
Percent elongation= (Lf-Li)/Li
Percent reduction in cross-sectional area=(Ai-Af)/(Ai)
Creep is the process by which a material elongates via plastic deformation under elevated temperature
conditions to allow thermally activated processes (such as diffusion) to participate in deformation response
o Assuming constant load, plot strain vs. time
o Creep is a problem for load bearing biomaterials, especially polymers