Download biocompatibility of materials implanted in vivo: definitions

3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
I
LECTURE 17: NANOMECHANICS AND BIOCOMPATIBILITY :
PROTEIN-BIOMATERIAL INTERACTIONS
Outline :
REVIEW LECTURE #16 : NANOMECHANICS OF CARTILAGE ............................................................. 2
BIOCOMPATIBILITY OF MATERIALS IMPLANTED IN VIVO: DEFINITIONS ......................................... 3
TEMPORAL BIOLOGICAL RESPONSES TO IMPLANTED BIOMATERIALS ......................................... 4
BLOOD-BIOMATERIAL INTERACTIONS ................................................................................................ 5
KINETICS OF PROTEIN ADSORPTION ................................................................................................. 6
USE OF STERIC REPULSION TO INHIBIT PROTEIN ADSORPTION ................................................... 7
THERMAL MOTION OF POLYMER BRUSHES : MOVIE ........................................................................ 8
POLY(ETHYLENE GLYCOL) AS A BIOINERT COATING ....................................................................... 9
Objectives:
To establish a fundamental qualitative and quantitative scientific foundation in
understanding the biocompatibility of biomaterials implanted in vivo
Readings: Course Reader Documents 29, 30
Multimedia : Polymer Brush Demos (posted on stellar)
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
REVIEW : LECTURE 16 NANOMECHANICS OF CARTILAGE
0
m)
Stiffness (MPa)
Force (nN)
Substrate
-Definitions; articular cartilage function and structure, proteoglycan, aggrecan, hyaluronan, link protein, collagen,
chondrocyte, glycosaminoglycan (GAG), chondroitin sulfate
- Loading Conditions : withstands ~3 MPa compressive stress and 50% compressive strain (static conditions),
equilibrium compressive moduli ~0.1-1MPa
- Composition : 80% HOH, collagen (50-60% solid content, mostly type II), aggrecan (30-35% solid content), hyaluronan,
~3-5% cartilage cells (chondrocytes)
(Seog+ J. Biomech 2005 in press, online &
Aggrecan Tip vs. Aggrecan Substrate
(Macro)
Dean, Han+ unpublished data 2005)
1.5
10
compressive
moduli : human
M
ankle cartilage
8
1
+ (approximate)
50% prestrain
6
(Nano)
0.5 Ag-Ag colloid
4
2
1M
1200
0.01M
0.1M
0.001M
0
0
0
0
400
800
1200
Distance (nm)
- Force vs. Distance converted into Stress,  
0.2
0.4 0.6
Strain
0.8
1
● stiffens nonlinearly with increasing strain at the molecular level→
mechanism to prevent large strains that could result in permanent
deformation, fracture, or tearing.
Lf
F
D
1 
1
, where As= surface interaction area vs. Strain  
As
Lo
ho (aggrecan)
 d 
where ho(aggrecan)= initial uncompressed height of aggrecan (zero force), Stiffness  

 d  0
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
Lecture 16 Nanomechanics of Cartilage: Definitions
articular cartilage : connective avascular (contains no blood vessels) tissue covering the ends of the bones in synovial joints that allow smooth,
low friction, painless motion
proteoglycan: A molecule that contains both a protein core and glycosaminoglycans, which are a type of polysaccharide. Proteoglycans are
found in cartilage and many other connective tissues.
aggrecan: the largest aggregating proteoglycan, found in cartilage tissue and the intervertebral disc, has a bottle-brush configuration composed of
a protein core backbone and densely spaces glycosaminoglycans
glycosaminoglycan (GAGs) : Polysaccharides containing repeating disaccharide units that contain either of two amino sugar compounds -- Nacetylgalactosamine or N-acetylglucosamine, and a uronic acid such as glucuronate (glucose where carbon six forms a carboxyl group). Also
called mucopolysaccharides in the older literature. GAGs are found in the lubricating fluid of the joints and as components of cartilage, synovial
fluid, vitreous humor, bone, and heart valves.
chondroitin sulfate (CS) : One of several classes of sulfated glycosaminoglycans that is a major constituent in various connective tissues,
especially in the ground substance of blood vessels, bone, and cartilage. Chondroitin sulfate is a sulfated glycosaminoglycan (GAG) composed of
a chain of alternating sugars (N-acetylgalactosamine and glucuronic acid). It is usually found attached to proteins as part of a proteoglycan.
CS→
← HA
hyaluronan (HA) : Hyaluronan (HA also called hyaluronic acid or hyaluronate) is an anionic polysaccharide composed of repeating disaccharides
of beta-1-4-glucuronate-beta-1-3-N-acetylglucosamine distributed widely throughout connective, epithelial, and neural tissues. The polysaccharide
appears to be unique amongst glycosaminoglycans as it is synthesized, and exists in vivo, without attachment to any protein. (As such, it is not
synthesized via the usual intracellular organelles that involve protein synthesis; rather, it is extruded from the cell membrane, catalyzed by the
enzyme hyaluronan synthetase.) It can be synthesized with a very large molecular weight (1,000 - 5,000 kDa). In cartilage, a globular domain at
the N-terminus of aggrecan, termed G1 or the hyaluronic acid binding region (HABR). binds to HA in an interaction that is stabilized by link protein.
link protein : A ~45 kDa globular protein that stabilizes the interaction between aggrecan and HA.
chondrocyte : cartilage cells responsible for the synthesis and maintenance of the extracellular matrix; chondrocyte-like cells are also found in
the intervertebral disc if the spine
collagen: The fibrous protein constituent of bone, cartilage, tendon, and other connective. Type II is the major fibril-forming collagen in cartilage.
(There are currently 28 known, distinct, collagen types.)
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
BIOCOMPATIBILITY OF MATERIALS IMPLANTED IN VIVO: DEFINITIONS
Biocompatibility : the ability of a material to perform with
an appropriate host response in a specific application, a
material that does not producing a toxic, injurious, or
immunological response in living tissue; no irritation,
inflammation, thrombosis, allergic reactions, coagulation,
cancer
contact lenses
intraocular lenses
dentures
• tissue engineering
scaffolds
• artificial organs
Bioinert - Biomaterials that elicit little or no host response,
in terms of nanomechanics a net zero (or close to zero)
surface potential with physiological environment,
sometimes called "non-fouling"
Interactive or Bioactive- Biomaterials designed to elicit
specific, beneficial responses, e.g. ingrowth, bioadhesion.
Bioadhesion : may be defined as the state in which two
materials, at least one of which is biological in nature, are
held together for extended periods of time by interfacial
forces. The biological substrate may be cells, bone,
dentine, or the mucus coating the surface of a tissue. If
adhesive attachment is to a mucus coating, the
phenomenon is sometimes referred to as
mucoadhesion. e.g. cell-to-cell adhesion within a living
tissue, wound dressing, and bacteria binding to tooth
enamel.
cochlear
implants
hip implant
heart
pump/
valves
bone
substitute
materials
drug
delivery
systems
knee implant
blood-contacting devices
(vascular grafts, stents)
Viable - incorporating live cells at implantation, treated by
the host as normal tissue matrices and actively resorbed
and/or remodeled
Biomaterial, Biomedical Materials : nonliving (artificial)
material intended to interact with a living (biological)
system, replacement for “broken” anatomical parts or
physiological systems
Biofilm : When a biomaterial is exposed a physiological
environment the resulting a complex aggregation that may
contain biomacromolecules, cells, bacteria, microorganisms
Examples of Biomaterials; medical implants, heart valves,
vascular grafts, contact lenses, drug delivery systems,
scaffolding for tissue regeneration, breast implant, hip joint
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
TEMPORAL BIOLOGICAL RESPONSES TO IMPLANTED BIOMATERIALS
• living materials respond rapidly to foreign materials (<1 s)
• new layer of protein coats (isolates) biomaterial surface (minutes)
• attachment of platelets, bacteria, yeasts, and additional proteins to surface (minutes-hours)
• alteration in cell and tissue behavior (minutes-years)
Host Effects :
Blood Clots and Thrombosis
Inflammatory Response
Immune Rejection
Fungal Infections and Diseases
Irritation and Inflammation
Fresh out of box
Biomaterial Degradation
Abrasive Wear
Fatigue
Stress-Corrosion Cracking
Absorption from Biofluids
(*chemical attack)
Mechanical Failure
after a few minutes
15 m
TappingMode AFM images in saline of the convex face of commercial
PHEMA soft contact lenses. Fresh "out of the box" contact lens (left)
displays scratches on the surface that originate from the mold during the
manufacturing process. The scratches are 5 to 10nm in depth and 150 to
850nm in width. Several large isolated features (130 to 250nm in height)
are also observed. The RMS roughness on the surface is 14nm. Used
contact lens (right) of the same exact type and brand in the left image. The
lens surface is coated with particulate adsorbates. A scratch-like feature is
visible running top to bottom in the image, and appears decorated with
contaminants. The RMS roughness on the surface is 30nm. Scan size for
both images is 15µm and z range is 160nm (Veeco, Inc)
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
BLOOD-BIOMATERIAL INTERACTIONS
Blood Compositions
and
Solution
Conditions :
pH7.15 - 7.35
IS=0.15 M
Temperature ~37°C
-cells and
(~45 wt.%)
BLOOD FLOW
blood plasma
proteins
PLATELETS!
D. Gregory http://medphoto.wellcome.ac.uk
http://www.rinshoken.or.jp/org/CR/photo-e.htm
platelets
The liquid portion of
the blood, the plasma
or serum (55 wt. %), is
a complex solution
containing more than
90% wate
- 6-8 wt.% proteins in
plasma (over 3,000
different
types),
including :
-58% albumins
-38% globulins
-4% fibrinogens
BLOOD PRESSURE+
ATTRACTIVE
FORCES
BLOOD CLOT!
-acute occlusive thrombosis
mobility,
denaturation,
- infection / inflammation
exchange, desorption
- neointimal hyperplasia
adsorbs
BIOMATERIAL SURFACE
solid-liquid Interface
bound water,
ions,
small molecules
2-10Å
-The majority of blood plasma proteins are net negatively charged. Each has its' own heterogeneous surface chemistry
and unique intermolecular potential with biomaterial surface that changes and evolves with time in vivo.
→ want bioinert surface
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
KINETICS OF PROTEIN ADSORPTION
Molecules can be brought to the surface by diffusion; (I. Szleifer, Purdue University)
z
U(D)
BIOMATERIAL
SURFACE
C
 2C
 D 2 ; C = concentration, D = diffusion coefficient, z = distance, t = time
t
z




2

 protein (z,t)
  protein (z,t)
U mf (z,t)  
 
 D


(
z,t)
 protein
 
2

t
z
z 
z
 
Ideal diffusion-controlled Kinetically activated regime, "driven" diffusion, 
controlled only by
regime

motion that arises from surface-protein
interactions
 density gradient

ρ protein (z,t)= local density of protein molecules
U mf (z,t)= net "potential of mean force" including protein - surface potential
 more complex theories take into account protein - protein interactions and protein
conformational changes
U mf (z,t) - can have many different components, both attractive (e.g. hydrogen, ionic, van der Waals, hydrophobic,
electrostatic) and repulsive (e.g. configurational entropy, excluded volume, osmotic, enthalpic, electrostatic, hydration),
can lead to complex interaction profiles, will change if conformation of protein changes
- Initial protein adsorption will be determined by longer range, larger spatial length scale of averaged surface properties
(e.g. average surface charge per unit area→EDL)
- Secondary stages of protein adsorption depend on shorter range biomolecular adhesive binding processes that take
place when the protein is in close contact with the surface (e.g. the conformation, orientation, and mobility of the adsorbed
proteins, the time scale of conformational changes, protein exchange and desorption, and interactions of adsorbed
proteins with each other).
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
USE OF STERIC REPULSION TO INHIBIT PROTEIN ADSORPTION
→ generally can't use charged surface EDL repulsion
as a mechanism to inhibit protein adsorption
(I. Szleifer, Purdue University)
→ one method: use steric repulsion of surface
functionalized (chemisorption, physisorption) with
polymers
Charged surface:
Hydrophobic surface:
proteins expose charged
groups.
proteins expose hydrophobic
patches.
e.g. human serum
albumin7
RP
adsorbed
proteins
(II.)
(I.)
Modes of protein adsorption:
(I.) adsorption of proteins to the top boundary of the
polymer brush
(II.) local compression of the polymer brush by a
strongly adsorbed protein
(III.) protein interpenetration into the brush followed by
the non-covalent complexation of the protein and
polymer chain
(IV.) adsorption of proteins to the underlying
biomaterial surface via interpenetration with little
disturbance of the polymer brush
RP
(III.)
(I.)
end-grafted
polymer brush
Lo
s
solid substrate
(IV.)
(Halperin, Langmuir 1999)
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
THERMAL MOTION OF POLYMER BRUSHES : MOVIES
(left) : (J. Marko (Cornell U) : http://www.lassp.cornell.edu/marko/thinlayer.html)
(right) : (*J. Hoh (John Hopkins U) : http://162.129.38.210/Hoh_Lab_Pages/Gallery.html)
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3.052 Nanomechanics of Materials and Biomaterials Thursday 04/19/07
Prof. C. Ortiz, MIT-DMSE
POLY(ETHYLENE OXIDE) AS A BIOINERT COATING
The most extensively used polymer for
biomaterial surface coatings:
(tgt)
- hydrophilic and water-soluble at RT
t
O
H
H
-forms an extensive H-bonding network;
intramolecular H- bond bridges between -Ogroups and HOH→ large excluded volume
-• locally (7/2) helical supramolecular
structure (tgt axial repeat = 0.278 nm)
O
O
H
O
t g tO t
t
O
t t
t t
H
g
0.278 nm
- - -- - - --
• steric (large excluded volume)
-high flexibility, molecular mobility
-low van der Waals attraction
•
electrostatic
double layer
forces
• neutral
However:
-poor mechanical stability
-protein adhesion reported under certain
conditions (long implant times)
-maintains some hydrophobic character
• hydrophilic/ water soluble :
hydration enthalpic penalties for
disruption of supramolecular
structure H-bonding with water
O
O
H
H
H
O
O
H
O
• high flexibility
& mobility :
no local steric or
charge
O
• neutrality : won’t attract oppositely charged species
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