Karla Brammer1 1Materials Science and Engineering, UC San Diego Nanomaterials Lecture Jan. 6th 2011
Cell nanostructures
Cells are typically 1-100m in size, but their interior and exterior contains many nanometer size objects.
Plays important roles in both intracellular transport, cell signaling, gene expression and cellular division.
Provides "tracks" with its protein filaments for transport of organelles, molecules, vesicles, etc
Element Microfilament F-actin Diameter (nm) 8-10
Intermediate filaments
Microtubules
8-10
25
Proteins are surrounded by a phospholipid (hydrophilic head and two hydrophobic tails) bi-layer.
They transduce information from the external environment, extracellular matrix (ECM), to the cell as well as reveal the status of the cell to the outside, allowing rapid and flexible responses to changes in the environment.
Motivation
Fabricating nano-features upon biomaterial surfaces provides features that are on the same scale as the bio-environmental features that interact with cells. Looking at what happens on different nanotextures can help to uncover signaling pathways that promote the desired cellular response.
Basic definitions
Nanomaterial Particles with lengths that range from 1 to 100 nanometers in two or three dimensions. ASTM E 2456-06 : Terminology for nanotechnology. ASTM International 2006 Biomaterial A non-living material intended to interface with biological systems to evaluate, treat, augment or replace any tissue, organ or function of the body. Chester 1986 Nanobiomaterial A biomaterial substrate composed of nanometer-scale components. Example : the inorganic bone matrix is comprised of apatite crystals with final dimensions 400 x 30 x 75 . Nanomedicine The monitoring, repair, construction and control of human biological systems at the molecular level, using engineered nanodevices and nanostructures.
Surface structures are often mobile Much higher surface/volume ratio for micro/nano
Nanobiomaterials have an even greater increase in number of atoms at their surface and possess a higher surface area to volume ratio than conventional microscale biomaterials.
Thus scientific developments of nanobiomaterials are multiple, from wound healing, bone implants to bone (or cartilage) tissue engineering scaffolds, to stem cell differentiation.
Discovery of Topography
The ability of the substratum to influence cell orientation, migration and cytoskeletal organization was first noted by Harrison in 1911.
Grew cells on a spider web. The cells followed the fibers of the web in a phenomenon called physical guidance. Observed variations in the behaviour of cells due to the solid support they grow on.
Later, in 1964, it was first proposed that cells react to the topography of their environment.
Surface shape and physical features themselves.
Since then, numerous studies have shown that many cell types react strongly to topography, especially on the nanoscale. The role of a substrate is more than merely providing mechanical support
Act as intelligent surfaces. Providing chemical and topographical signals. Guides and controls cell behavior.
Incorporating topographical consideration into the design of scaffold environments is becoming increasingly important in light of these studies. With advances in nanofabrication technologies the promise of and the unknown information about topographical effects in manipulating the cell-substrate interaction is now being uncovered
Inspiration
New fabrication technologies and new nanotechnologies have provided biomaterial scientists with enormous possibilities when designing customized tissue culture supports and scaffolds with controlled nanoscale topography. The main challenge:
To effectively design these scaffold for specific tissue engineering applications. Choose the appropriate combination of biomaterial composition, size, structure, etc. Be able to tailor towards applications as challenging and complex as
(a) wound healing (b) new bone growth and orthopedic implant technologies (c) stem cell differentiation (d) reconnect nerves/spinal cord injuries/brain damage
Nanopattern fabrication
The definition of nano-structures on the substrates relies on the clean room lithographic and Si processing methods. Lithography
Usually, a computer-designed pattern is exposed by means of light, electrons, ions or imprinted. Carried out on a special light or electron-sensitive material or imprinted into a special deformable polymer, which is then used in subsequent pattern transfer processes as a mask, or, alternatively, used for cell culturing as it is.
Contact guidance
Controlling cell adhesion, orientation and morphology through topographical patterning is a phenomenon that is applicable to a wide variety of medical applications such as implants and tissue engineering scaffolds.
Wound healing Neuron alignment
Effects on cells based on ridge features. Sharper angles orient cells better. Cells adhere better when aligned with surface texture. Possible explanations:
Higher surface energy leads to preferential protein binding Alignment of adhesion plaques Abrupt edges lead to alignment or bridging Stochastic spreading of cells
Cell elongation and alignment on grooves and ridges of nanoscale dimensions was compared with the morphology and orientation of cells cultured on smooth substrates. It was found that ridges 70 nm wide induced human corneal epithelial cells to elongate and align along the topographic features. Valuable for the development of implantable prosthetics.
Mechanotransduction
Cells are exquisitely sensitive to forces of varying magnitudes, and they convert mechanical stimuli into a chemical response. Integrincytoskeleton linkage is sensitive to force.
Close-up of a focal adhesion showing the balance of external and internal forces (Fext and Fcell, respectively) in driving stress at a mechanosensor. Depicted are actin stress fibers (red) anchored into focal adhesions (multicolored array of proteins) that bind to the ECM (blue) through integrins (brown).
This balance of forces provides the stress necessary for mechanical sensing. This triggers a cascade of reactions that alter the balance of anabolic / catabolic events within the cell. This is a major regulator in cell homeostasis and development.
Demonstrated the use of nanoscale disorder to stimulate mesenchymal stem cells to produce bone mineral in vitro. Claimed that the focal adhesion complexes determined by the nanostructure effected the differentiation pathways.
Increased Osteogenic Gene Expression
Osteocalcin (OCN)
Surface nanotopography directly induces pronounced changes to cell shape, and consequently gene expression, which can potentially mediate differentiation of stem cells into various cell types.
Cell Signaling Stress fibers
Ceramic nanopores/nanotubes
Electrochemical anodization
Anodic Aluminum Oxide (AAO) TiO2 Nanotubes
Controllable features
Film thickness Pore size/diameter Wall thickness Interspaces
Demonstrated:
An increase in the alkaline phosphatase activity, bone forming ability on nanoporous surface. Enhanced matrix production of MSCs when they are cultured on nanoporous alumina substrates.
Osteoblasts (bone cells) Higher density of cells. 3-4X accelerated growth. Up-regulated mineralization. Intricate interaction.
Reduction of ECM
Nanofiber designs
Electrospinning technique:
Nanofibrous scaffolds are promising candidate scaffolds for cell-based tissue engineering. More closely mimics the niche-like unit (ECM fibrils) for facilitating proper cell behavior.
Electrospinning generates loosely connected 3D porous mats with high porosity and high surface area which can mimic ECM. Can be randomly or aligning on the surface based on ground collection plate
Nanofiber advantage
Scaffold architecture affects cell binding and spreading. The cells binding to scaffolds with microscale architectures flatten and spread as if cultured on flat surfaces. The scaffolds with nanoscale architectures have bigger surface area for absorbing proteins and present more binding sites to cell membrane receptors. The adsorbed proteins further can change the conformations, exposing additional binding sites, expected to provide an edge over microscale architectures for tissue generation applications.
General remarks
Toxicity must be avoided but inertness is not a high priority in nano/biomaterials.
Bioactivity: positive interaction and effects on the human body.
Metallics, alloys and ceramic biomaterials are useful for hard tissue (bone) but not suitable to replace soft tissues because of markedly different mechanical properties.
Must match the material properties to the body material
Consider modulus, shear, wear particles, fatigue, strength Applicable for 3-D geometries
Conventional polymers are used for many of todays pliable disposable or biodegradable medical devices. New functional biomaterials are anticipated
Questions