Conducting microhelices from self-assembly of protein fibrils
Outline of today’s class: STRESS-SUPPORTING STRUCTURES...Gibson and Kenedi Banding visible on...
Transcript of Outline of today’s class: STRESS-SUPPORTING STRUCTURES...Gibson and Kenedi Banding visible on...
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Outline of today’s class:STRESS-SUPPORTING
STRUCTURES1. Tissues and organs differ greatly in
deformability even though made up of few components.
2. Collagen fibers: Orientation patterns in different organs match mechanical function.
3. Collagen fibers: Periodic structure of crystalline fibers at different scales.
4. Elastin fibers are amorphous and rubber-like.5. Glycosaminoglycans (GAGs) are
polysaccharides that keep water inside tissues.
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1. Tissues and organs differ greatly in compliance (deformability).
tendonneck ligamentskin
Stress-strain curves (constitutive relations) for various tissues and organs:
stress
strain
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Are the basic rules of linear elastic mechanics obeyed by most tissues?
• Matter is a continuum.• Linearity. Directly proportional relation
between stress and strain. No terms with exponent higher than 1.
• Elasticity. Time-independent mechanical behavior. Loading and deformation independent of time.
• Isotropy. The elastic constants are independent of loading direction.
• Conservation of mass.
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Tissues and organs differ greatly in deformability due to:
•crystalline vs. amorphous fibers(collagen vs elastin fibers)
•fiber reinforcement(tendon vs liver)
•crosslinking (newborn vs aged)
•molecular configuration(collagen vs GAG)
•ceramic content •(bone vs cartilage)
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Three basic structural families of macromolecules: Collagens, elastins and
polysaccharides (GAGs)
•Diversity in mechanical behavior among tissues due primarily to variations in the organization of fibers of these macromolecular substances.
•Model of supporting tissues as fiber-reinforced composite materials. A soft, aqueous GAG "matrix" gel is reinforced with fibers of collagen and elastin. [Bone is further reinforced with ceramic particles.] The model illuminates the critical importance of fiber orientation on stiffness and strength of tissues.
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2. Collagen fibers: Orientation pattern in various organs
match mechanical function.• Tendons. Thick fibrous bundles that connect muscle
to bone. Support axial forces. Collagen fibers are uniaxially oriented.
• Ligaments are similar to tendon in structure but connect bone to bone.
• Cornea. Membrane protects curved eye surface from tangential forces. Planar orientation of collagen fibers.
• Dermis (skin). Collagen fibers nearly randomly oriented (some orientation in plane of epidermis).
• Articular cartilage. Membrane between apposed bones in joints. It “lubricates” joints. “Cathedral”architecture of collagen fibers.
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Tendon connects muscle to boneand supports axial forces
Image removed due to copyright restrictions.Diagram of human arm bone, muscle and tendon structure, showing elbow flexion and extension.
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Quaternary structure of collagen. Fibers are probably single crystals, about 100 nm diameter, with a 64-nm periodicity (“banding”).
Image removed due to copyright restrictions.Electron microscope photograph of collagen fibers.
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Tendon fibers
200 μm
What is the orientation of 100-nm thin collagen fibers in 200-μm thick tendon fibers? Do they form helix round axis of tendon fiber? Or is their axis parallel to tendon fiber axis?
Image removed due to copyright restrictions.Electron microscope photograph.
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Effort to identify orientation of 100-nm collagen fibers by dissolution in aqueous acetic acid. However, collagen fiber orientation is not clearly shown in this experiment.
100 μm
Image removed due to copyright restrictions.Electron microscope photograph.
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Effort to identify orientation of collagen fibers by fracture. Wet tendon fibers fracture in “fibrillar” mode. 100-nm collagen fiber orientation not shown.
40 μm
Image removed due to copyright restrictions.Electron microscope photograph.
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Fracture surface of dry tendon
20 μm
Try a different approach. Dehydrate first, and then fracture tendon.
Image removed due to copyright restrictions.Electron microscope photograph.
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Detail of fracture surface for dry tendon showing parallel arrangement of fibers along major fiber axis. Individual fibers,about 100 nm diameter, are single crystals of collagen.
2 μm normal tofracture surface
Research question: how are collagen crystals bound to each other?
Image removed due to copyright restrictions.Electron microscope photograph.
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The eye
Conjunctiva
THE HUMAN EYE
Aqueous HumourLens
PupilOptic Nerve
Retina
Fovea
Cornea
Iris
Zonula
Figure by MIT OpenCourseWare.
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Tadpole cornea
CORNEA. Membrane protects curved eye surface from tangential forces. Planar orientation of collagen fibers.
Research question: how can the cornea, a two-phase material, be transparent?
Image removed due to copyright restrictions.
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Fish cornea
Image removed due to copyright restrictions.
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Detailedview of skin
Image removed due to copyright restrictions.Medical illustration (by Frank Netter) of the structure of human skin.
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SKIN
Figure by MIT OpenCourseWare.
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A, arteriolesN, peripheral
nerveAlmost all else is quasi-randomassembly ofcollagen fibers
Dermis
100 μm______
Histology photo removed due to copyright restrictions.
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_____20 μm
DERMIS (Skin). Collagen fibers nearly randomly oriented (there is some orientation in plane of epidermis). Supports forces along all three axes.
Microscope photo removed due to copyright restrictions.
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Collagen fibers in human dermis: 4 magni-fications
Gibson and Kenedi
Bandingvisible onindividual fibrils(quaternary structure)
Microscope photos removed due to copyright restrictions.
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Skin extensibility varies over right side of chest. Langer’s lines coincide with axis of minimum extensibility.
right nipple
Diagram removed due to copyright restrictions.
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normal dermis
scar
light scattering pattern shows axial orientation for scar
Image removed due to copyright restrictions.
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Regeneration of the dermis in guinea pig skin wounds. Studied by histology (up) and laser light scattering (down).
Image removed due to copyright restrictions.
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Orgill, 1983
Regenerated dermis(guinea pig)
Scar (guinea pig)
Orgill, MIT Thesis,1983
Research question: How does scar form?
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ARTICULAR CARTILAGE. Membrane between apposed bones in joints (arrow). It “lubricates” joints. Does not “damp”impact forces.
Photo removed due to copyright restrictions.
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ARTICULAR CARTILAGE. It “lubricates” joints partly through a “weeping” mechanism in which synovial fluid is squeezed out of cartilage during compression.
Diagram removed due to copyright restrictions.
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ARTICULAR CARTILAGE. Collagen fiber orientation varies greatly from surface to interior. Fiber orientation resembles arcing struts in cathedral architecture.
Research question: How does cartilage maintain the lowest known coefficient of friction?
Histology image removed due to copyright restrictions.
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Abnormally stretchy skin
Three photos removed due to copyright restrictions.
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From Sci. Americancervix
a baby inside the uterusabout to be born
Diagram removed due to copyright restrictions.
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during childbirth
after
before
time, s
deformation
From Harkness and HarknessViscoelasticity of uterine cervix
Graph removed due to copyright restrictions.
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Chicken with normal bones
Collagen fibers in bones lackcrosslinking
X-ray images removed due to copyright restrictions.
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3. Collagen fibers: Periodic structure of crystalline fibers at
different scales.• Protein fibers typically comprise a hierarchy
of "ordered" structures; each member of the hierarchy is characterized by repetititon of a structural motif (periodicity) either along the macromolecular chain or across a collection of chains.
• Primary structure is the complete amino acid sequence of each protein chain (no information on configuration).
• Secondary structure is the local 3-D configuration of a few (3-5) amino acid units (residues) along the chain (helix?)
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3. Collagen fibers: Structure at different scales (cont.)
• Tertiary structure is the configuration of the entire molecule (triple helix).
• Quaternary structure refers to the crystalline structure formed by aggregation of many molecules ("banded" fiber). Crystalline structure of fibers restricts stretching to about 10% along axis.
• Architecture of fibers refers to orientation of fibers that comprise the macroscopic tissue or organ.
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primary
secondary
tertiary
quaternary
Hierarchy of collagen structures
Diagram removed due to copyright restrictions.
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Quaternary collagen structureobserved as banding with a 64-nm period
100 nm
_________Research question: How does collagen banding induce clotting?
Image removed due to copyright restrictions.
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When collagen melts it becomes gelatin, an amorphous polymer
What is the water content in jello?
Chemical diagram removed due to copyright restrictions.
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4. Elastin fibers are amorphous and rubber-like
• Primary structure of this protein comprises mostly nonpolar amino acids (AA); weak forces between neighboring AA along chain (intramolecular interactions) as well as between AA in adjacent chains (intermolecular).
• No structural periodicity detected; therefore, no hierarchy of periodic motifs observed.
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ElastinX-ray diffraction pattern is negative(amorphous)
Image removed due to copyright restrictions.
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P, papillary dermis
R, reticulardermis
Elastin fibers stain black
SKIN
100 μm________
Histology photo removed due to copyright restrictions.
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4. Elastin fibers (cont.)•No interactions. There is no crystallinity in elastin fibers (amorphous). Its macromolecules are free of interactions (either intra- or intermolecular) and are, therefore, capable of stretching out their coiled configurations all the way, until covalent bonds are stretched. •Mechanical behavior. Rubber-like behavior, capable of extension by 150%, is observed; however, unlike tire rubber, elastin is a liquid-filled elastomer (it would make a terrible tire rubber). Amorphous elastin fibers in contrast to crystalline structure of collagen fibers (compare relatively stiff, crystalline Nylon fibers to stretchy, amorphous Spandex fibers).
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4. Elastin fibers (cont.)•Neck ligament: free and easy movement of head around spinal column.•Aorta: both collagen and elastin fibers reinforce blood vessel wall. Fibers in vessel wall become enriched in elastin the closer is vessel to the heart (increased distance from periphery); near heart, vessel wall becomes more stretchy and partly damps mechanical energy, probably contributing in converting the on-off pulsatile energy output of the heart to a more continuous flow pattern. •Skin. Effects of ageing on stress-strain curve of skin. Loss of elastin?
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Aorta. Data on% elastin (as % elastin+collagen) in this major blood vessel.Elastin content decreases from top (left)through chestdiaphragm (right).
dog
elephant
hippo
giraffe
Four graphs removed due to copyright restrictions.
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Aorta. Data on% elastin (as % elastin+collagen) in this major blood vessel.Elastin content decreases from top (left)through chestdiaphragm (right).
dog
elephant
hippo
giraffe
Four graphs removed due to copyright restrictions.
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5. Glycosaminoglycans (GAGs) in tissues are polysaccharides.
•Repeat units: Macromolecular chain made up of sequence of sugar, not AA, units. GAGs are side chains of “core” protein chain: proteoglycan. •Sequence of fixed negative (electrostatic) charges along chain axis stiffens it up due to repulsive interactions among charged ions. Highly ionic macromolecular network swells highly in water, occasionally increasing its dry mass by over 1000X. About two-thirds of body weight is water, most of it "trapped" by GAGs. (Proteins also immobilize water.)
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5.Glycosaminoglycans(GAGs)(cont.).
•Swollen GAGs serve as the soft "matrix" in the mechanical model of supporting tissues as fiber-reinforced composites. This matrix is stiffened and strengthened by collagen and elastin fibers. •Stiffness of tissues with very high GAG content depends strongly on maintenance of ionic charges at "full strength". E.g., stiffness of articular cartilage drops by about 50% when the salt-free water that swells it is replaced with a high-salt water solution. Salt ions "screen" fixed charges on GAG chains, causing drop in repulsive forces among them. (Research question: What accounts for remainder 50% of stiffness of cartilage?)
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SummaryTissues differ in deformability. They
comprise fibers and gel-like matrix. Orientation of crystalline collagen fibers in
tissues roughly relates to mechanical function of tissue.
Hierarchy of structural order (periodicity) in collagen and other proteins.
Elastin fibers are amorphous and rubber-like.Glycosaminoglycans (GAGs) are electrically
charged polysaccharides. Charges retain much water. Charge repulsion straightens up chains.
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MIT OpenCourseWarehttp://ocw.mit.edu
20.441J / 2.79J / 3.96J / HST.522J Biomaterials-Tissue InteractionsFall 2009
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