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Body Synthetic - Tissue Engineering
Stem Cells and Scaffolds
166
Other
Undergraduate 2
08/17/2014

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Term
reasons for tissue engineering
Definition
transplantation crisis, shortage of donor organs and tissues available
transplant rejection
Term
what are the four basic tissue types
Definition
muscle tissue, nerve tissue, epithelial tissue, and connective tissue
Term
muscle tissue types and functions
Definition
skeletal (voluntary), smooth (involuntary), cardiac (only found in heart, involuntary)
contracts for movement and support
Term
nerve tissue: types and functions
Definition
brain, spinal cord, nerves, gives signals to muscles, informs us of environmental conditions
Term
epithelial tissue: types and functions
Definition
lines our organs such as stomach lining, skin (epidermis) (endothelial - blood vessels)
Term
Three Essential Components of Tissue
Definition
cells, extracellular matrix, soluble factors
Term
soluble factor examples
Definition
growth factors, hormones, cytokines, soluble gases
Term
cellular tissue engineering components
Definition
cells, scaffolds, extracellular signals = biological substitute
Term
example of natural scaffold
Definition
collagen
Term
a method to promote blood supply structure
Definition
silicone chip
Term
what prevents blood from coagulating in blood vessels?
Definition
endothelial cells
Term
why don't scaffolds work well in vascular tissue engineering?
Definition
smooth muscle is more easily growing than endothelial cells, scaffold gets covered with smooth muscle, and blood coagulates at the site
Term
can neurons divide/proliferate?
Definition
no
Term
how do you repair axons?
Definition
make the environment conducive to regeneration
Term
three layers of the skin
Definition
epidermis - top layer
dermis
hypodermis
Term
epidermis contents
Definition
top layer, not much ECM, densely packed
Term
dermis contents
Definition
mostly ECM, fibroblasts
Term
fibroblasts
Definition
cells that create rapid growth, often scar tissue
Term
negative effects of fibroblasts in the heart
Definition
heart attack, tissue dies forever, fibroblasts create scar tissue, difficult to contract
Term
how do you know if you damaged the dermis?
Definition
you bleed
Term
formation of ECM - dynamic or static?
Definition
dynamic!
Term
how do cells know to attach to a scaffold?
Definition
growth factors, cells will follow a growth factor gradient
Term
our ECM is hydrophillic, so why don't we dissolve?
Definition
strands are tangled and interwoven, no opportunity to dissolve
Term
connective tissue: types and functions
Definition
connects, supports, and protects other tissues, ligament, bone, tendon, blood, hair
Term
tissue engineering process
Definition
cell sourcing
cell expansion and manipulation
mechanical and molecular signalling
cell seeding and ECM expression
implantation of construct
Term
what was with that vacanti mouse?
Definition
they grew a human ear on the mouse's back
Term
why wouldn't the ear from the vacanti mouse function if implanted onto a human?
Definition
mouse epidermis would cause rejection -- xenograph (across species)
cartilage cells were from a cow - very different from human cartilage
Term
renewing (labile cells)
Definition
multiply constantly through life, high rate of death
Term
examples of renewing, labile cells
Definition
skin, intestinal epithelium, bone marrow
Term
expanding cells (stable)
Definition
low rate of death and replication, divide following stimulation
Term
static (permanent) cells
Definition
lack any capacity to divide
Term
stratum basale
Definition
new keratinocytes divide here and as they reproduce and make a structural protein called keratin, they are pushed upward until they die and form the outer skin layer
Term
Anatomy of the small intestine
Definition
crypt progenitors
enterocytes
goblet cells
villi
epithelial sheet
Term
crypt progenitors function
Definition
cells at bottom of wells in submucosa, divide every 12-16 hours, 200 cells per crypt every day
Term
enterocytes function
Definition
absorb nutrients
Term
goblet cells function
Definition
provide a mucus lining in the small intestine - keeps the intestine from digesting itself
Term
how long do cells live in the intestine?
Definition
cells reach the top of the villus and shed off in 5 days
Term
paneth cells
Definition
in the small intestion - at the crypt bottoms
secrete antimicrobial peptides and enzymes
Term
bone marrow
Definition
in marrow cavity of bone, synthesizes blood
Term
erythrocytes
Definition
blood cells - plasma life of about 120 days
Term
examples of expanding (stable) cells
Definition
endothelium, fibroblasts, liver cells, smooth muscle cells
Term
examples of renewing (labile) cells
Definition
skin, intestinal epithelium, bone marrow
Term
examples of static (permanent) cells
Definition
heart muscle cells, neurons
Term
cell sources
Definition
autologous
allogeneic
xenogeneic
Term
autologous cells
Definition
from the patient
differentiated cells of same or other tissue type
stem cells (e.g. from bone marrow, fat of other tissue, or saved from umbilical cord
Term
allogeneic cells
Definition
from other human sources
differentiated cells of same or other tissue type
fetal stem cells
embryonic stem cells
Term
xenogeneic cells
Definition
from a different species
differentiated cells of same or other tissue type
fetal stem cells
embryonic stem cells
Term
autologous cells - pros and cons
Definition
pros: immunologically acceptable
cons: not readily available, donor site morbidity
Term
allogeneic cells - pros and cons
Definition
pros: can be readily available
cons: not always immunologically acceptable
Term
xenogeneic cells - pros and cons
Definition
cons: requires engineering immunological tolerance, potential animal virus transmission
Term
stem cell definition
Definition
self renewal, reproduce itself
differentiation, differentiate into functional phenotypes
Term
symmetric division of stem cells
Definition
one stem cell divides into two stem cells
Term
asymmetric division of stem cells
Definition
one stem cell divides into one stem cell and one progenitor cell
Term
progenitor cells
Definition
can divide many times, can divide 2^30
Term
is stem cell division rate high or low?
Definition
low - progenitor cells do most of the division
Term
what is the ratio of stem cells to all cells?
Definition
1 in 10 to 15 thousand cells
Term
what do you use to expand cells?
Definition
culture flask - flat bottom
Term
contact inhibition
Definition
when cells come in contact with each other, they stop proliferating
this is generaly defective in cancer cells
Term
fibronectin
Definition
glycoprotein in the ECM - binds to integrin proteins
Term
how do you spur on the correct cell differentiation?
Definition
correct environment - nerve cells need electrical stimulation, bone cells need mechanical stimulation
Term
cellular construct
Definition
cells and scaffold structure that can be implanted into the body
Term
scaffold degradation
Definition
must be slow enough to allow tissue infiltration - if it degrades too fast, tissue will lose its structure
Term
cartilage cells - why should you stent it?
Definition
cells not mature enough in the first 12 weeks to counter the forces by the skin in healing - it would lose shape
Term
why can't you put chondrocytes (cartilage cells) in the center of a scaffold?
Definition
necrosis due to lack of nutrition
Term
necrosis vs apoptosis
Definition
cell homicide vs cell suicide, respectively
Term
leukemia
Definition
blood disease, not able to grow new healthy RBC - need a bone marrow transplant
generally, bone marrow injected into the blood stream directly
Term
totipotent
Definition
stem cells that have the potential to become any cell type in the adult animal body, and any cells of the extra embryonic tissue (placenta and umbilical cord)
Term
pluripotent
Definition
the potential to differentiate to all somatic cells (but not to those of the placenta and umbilical cord which is derived from a trophoblast)
Term
trophoblast
Definition
stem cell that placenta and umbilical cord are differentiated from
Term
multipotent
Definition
stem cells that can only differentiate into a limited number of type (ex. trophoblasts)
Term
embryonic stem cells
Definition
pluripotent, derived from inner cell mass of blastocyst
Term
induced pluripotent stem cells
Definition
also known as iPS cells, adult cells that have been genetically programmed to an embryonic stem cell-like state
Term
adult stem cells
Definition
undifferentiated cells that are found in differentiated adult tissues
ex) bone marrow, fat,blood,brain, and spinal cord, dental pulp, liver, skeletal muscle, pancreas, epidermis, mucosa of the digestive system
Term
fetal stem cells
Definition
cord blood, amniotic fluid and placenta
Term
source of a totipotent stem cell
Definition
4 cell stage of a zygote - all four totipotent - after thi step, cells are pluripotent
Term
source of pluripotent stem cells
Definition
inner cell mass of blastocyst
Term
embryonic stem cells - pros
Definition
Pros: able to differentiate into all tissues in the adult body
able to replicate indefinitely while retaining their undifferentiated pluripotent state
ease of purification
Term
embryonic stem cells - cons
Definition
risk of teratoma formation - insufficient differentiation
material of animal rejection - pathogen transmission
immunologic rejection
ethical concerns
Term
ESC - teratoma
Definition
encapsulated tumor with tissue or organ components resembling normal derivatives of all three germ layers
undifferentiated ESCs can form teratomas after implantation
Term
what kind of cells can serve to make iPS cells
Definition
fibroblasts - all have the same genome - environmental factors and gene expression determine function
Term
true/false - it is difficult to purify adult stem cells
Definition
true
Term
where do trophoblasts reside?
Definition
outer layer of the blastocyst
Term
morula
Definition
mass of cells in the inside of the blastocyst
Term
aintainense of pluripotency of ESCs
Definition
feeder cells - fibroblasts, and serum proteins
remval of feeder cells, ESCs spontaneously differentiate into all three germ layers
Term
hematopoietic stem cells
Definition
give rise to all blood cell types - white blood cells (leukocytes), red blood cells (erythrocytes), and platelets (thrombocytes)
Term
mesenchymal stem cells (MSCs)
Definition
give rise to a variety of cells from mesodermal lineages including chondrocyte, osteoblasts, fibroblasts, muscle cells, tendons, adipocytes (fat cells), endothelial cells
Term
endothelial progenitor cells
Definition
give rise to endothelial cells
Term
bone marrow transplant procedure
Definition
destroy diseased BM by chemotherapy and/or radiation
infusion of healthy BM into patients bloodstream
BM migrates into large cavities of the bones, begins producing normal blood cells
genetic makeups must match
Term
what kind of cells do you use for a bone marrow transplant?
Definition
hematopoietic cells
Term
three germ layers
Definition
endoderm, ectoderm, mesoderm
Term
mesenchumal stem cells - properties
Definition
immunoprivileged
do not express immunologically relevant cell surface markers
inhibit the proliferation of allogeneic T-cells in vitro an elicit no immune response after allogeneic or xenogeneic tranplantation
readily isolated from various sites of the human body, especially from BM and adipose tissues
expand in culture without differentiation fr 30-40 divisions
Term
fetal stem cells
Definition
amniotic fluid and placenta
cord blood
Term
amniotic flid and placental stem cells give rive to:
Definition
hematopoietic, mesnchymal, and pluripotent cells
Term
cord blood stem cells give rise to:
Definition
hematopoietic, pluripotent
Term
fetal stem cells, pros and cons
Definition
pros: ease of procurement, abundant source, higher capacity to proliferate, less immunogenic than adult stem cells, no ethical concerns
cons: limited number of cells, immunogenic
Term
why can't we use the stem cells we have saves in research labs now
Definition
they are garbage because we mixed crap with them and they are unusable now
Term
modulation of extracellular environment
Definition
growth factors, hormone, extracellular matrix components
Term
modulation of intracellular environment through gene transfection
Definition
induce gene expression: gene transfection of transcription factors through DNA delivery
silence gene expression SiRNA delivery
Term
conductive signal for regulation
Definition
scaffolds for supporting host tissue growth, ECM molecules
Term
inductive sigal for regulation
Definition
drug delivery of soluble factors
Term
biomaterias scaffolds
Definition
mimic ECM to support:
cell adhesion
cell differentiation
cell proliferation
Term
Scaffolds: requirements
Definition
large surface area to volume ratio
macroporous
biocompatible
mechanical properties
specific 3D shape
physical guidance or patterning: topographic cues
degrade at the same rate as tissue deformation
ability to incorporate drug releasing component
Term
porosity
Definition
how many pores through a certain surface area, also how large the pores are - different sized pores for different cells
Term
mnufacturing porous scaffold
Definition
solvent casting and particulate leaching
gas forming
freeze drying
rapid prototyping
hydrogel
Term
fibrous scaffolds
Definition
electrospinning
Term
solvent casting and particulate leaching scaffold technique
Definition
salt particles, casting, vacuum dry, immerse in water
extesively used for PLLA and PLGA scaffolds
pros: pore size and porosity can be tuned by changing the particle size and the polymer/particle ratio
cons: limited thickness
cytotoxic organic solvent
Term
why are PLLA, PGA, and PLGA used commonly?
Definition
FDA approved
Term
why are PLLA PGA, and PLGA not favored by tissue engineers?
Definition
hydrophobic, not great for applications
Term
gas forming - scaffold technique
Definition
gas creates the pores
compression molding using a heated mold
high pressure CO2 gas
gas dissolved in the polymer
closed pore morphology
pros: no organic solvents
cons: excessive heat prohibits incorporation of temperature labile materials into the polymer matrix
pores do not form an interconnected structure
Term
freeze drying - scaffold technique
Definition
polymer dissolved in aqueous solution
rapid cooling create phase separation
solvents removal by sublimation under vacuum
pros: no organic solvents and high temperature
cons: porosity and pore size difficult to control
Term
rapid prototyping
Definition
CAD design, 3D printing, three dimensional printing og scaffolds and cells
pros: predefined properties, interconnetivity
cons: slow slow slow
Term
hydrogels
Definition
water swollen cross-linked polymeric structures either by covalent bonds or entanglements - hydrogen bonds
Term
hydrogels posses a degree of flexibility very similar to natural tissue BECAUSE
Definition
of their significant water content
Term
hyaluronic acid
Definition
chemical cross linking in hydrogel
Term
agarose
Definition
physical entanglement of fibers
Term
alginate hydrogel
Definition
Ca2+ crosslinking
Term
how can you tell a cell is pluripotent?
Definition
it can generate a cell from each germ layer
Term
mesoderm layer
Definition
connective tissue
Term
endoderm layer
Definition
internal organs, etc.
Term
ectoderm
Definition
skin, nervous system
Term
adipocytes
Definition
fat tissue
Term
use of transcription factors to do what?
Definition
maniulate gene expression
Term
true/false - cells will not respond to injection of transcription factors into the media
Definition
TRUE
Term
how do you properly introduce transcription factors?
Definition
gene delivery
Term
how are scaffolds without cells advantageous?
Definition
no rejection, increased shelf life
Term
PLLA vs PGA
Definition
PLLA degrades slower, PGA degrade faster, PLGA is a copolymer
Term
disadvantages of PLLA and PGA
Definition
hydrophobic, only soluble in organic solvents
Term
fibrous scaffolds - technique
Definition
high voltage applied to polymer solution
electrostatic repulsion counteracts the surface tension
at a critical point, a thin fibrous stream is ejected
solvent evaporation
Term
electrospinning - control factors
Definition
fiber diameter, porosity, and morphology can be controlled by applied voltage, viscosity, solution conductivity, and temperature
Term
fibrous scaffolds - traits
Definition
large surface area:volume ratio
high porosity
add strength to composites
tubular (nerve, blood vessel, intestine
solid
Term
fibrous scaffolds, application
Definition
wound repair, sutures
skin tissue engineering
cartilage tissue engineering
cardiovascular
Term
mechanical properties of scaffolds
Definition
composed of polymers, ceramics, or composites
select material that closely resembles properties of tissue it is to replace
Term
soft tissue - material used
Definition
natural and synthetic polymers
Term
hard tissues - material used
Definition
metals, ceramics, composites
Term
glycosaminoglycans (GAGs)
Definition
unbranched polysaccharide chains composed of repeating disaccharid units
Term
glycosaminoglycan examples
Definition
hyaluronic acid, chondroitin sulfate (cartilage), dermatan sulfate, heparan sulfate, keratan sulfate
Term
GAGs are generally attached to _______ to form __________
Definition
ECM proteins; proteoglycans
Term
proteoglycan
Definition
GAG chains bound to a core protein
Term
collagen role in ECM
Definition
tensile strength
cell binding domain
Term
elastin role in ECM
Definition
resiliency and extensibility
Term
GAG role in ECM
Definition
attract water
resistance to pressure
Term
why do GAGs attract water?
Definition
negative surface attracted positive ions, creates a concentration gradient, osmotic forces pulls water to the more concentrated areas around the GAGs
Term
proteoglycan role in ECM
Definition
attract water, keep ECM cells hydrated, store growth factors, compressive strength
Term
glycoproteins role in ECM
Definition
binding domain for cells and ECM molecules
Term
what components of the ECM promote mechanical strength?
Definition
collagen, elastin, proteoglycans
Term
what components of the ECM promote cell binding?
Definition
collagen and glycoproteins
Term
two most important glycoproteins
Definition
fibronectin, laminin
Term
fiber forming ECM molecules
Definition
collagen and elastin
Term
space filling ECM molecules
Definition
glycoproteins and proteoglycans
Term
integrin
Definition
transmembrane receptor on cells for binding ECM proteins
Term
ligand
Definition
cell binding site on an ECM molecule
Term
RGD
Definition
cell attachment site of many adhesive proteins, Arg - Gly - Asp combination of nucleic acids
Term
YIGSR
Definition
cell adhesion nucleic acid sequence
Term
KRSR
Definition
enhances osteoblast adhesion but not endothelial cells or fibroblasts
Term
REDV
Definition
endothelial cells
Term
vascular endothelial cell growth factor
Definition
VEGF, blood vessel formation
Term
fibroblast growth factor family
Definition
FGF, cell division, angiogenesis
Term
Nerve growth factor
Definition
NGF, neuron survival, neurite extension, neuron migration
Term
transforming growth factor beta
Definition
TGF-Beta, wound healing
Term
bone morphogenetic protein
Definition
BMP, bone deposition
Term
double emulsion
Definition
hydrophobic nanoparticles/microspheres aqueous solution dropped into an organic solvent. Drops are collected from mixture and dropped into another solution, emulsifier
Term
hydrophobic nanoparticles/microspheres pros and cons
Definition
pros: controlled release rate, small or large particles, high entrapment proficiency
cons: inflammatory, organic solvents, potential denaturation, low drug loading efficiency for hydrophillic drugs
Term
entrapment efficiency
Definition
ratio of weight of drug entrapped int a carrier system to the total drug added
Term
loading efficiency
Definition
ratio of the weight of the drug to the weigt of the total carrier system (drug plus polymer carrier)
Term
heparin binding growth factors
Definition
bFGF, GDNF, NGF, NT-3, PDGF, Sonic Hedgehog
Term
affinity based drug delivery pros and cons
Definition
pros: high drug entrapment efficiency, water based, stable sustained release
cons: can only be used for growth factors that have binding affinity for heparin
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