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Tissue Eng > Tissue Eng Midterm > Flashcards

Tissue Eng Midterm Flashcards

1
Q

What is TE?

A

Application of engineering and life science methods and principles toward the development of biological substitutes that restore, maintain, or improve tissue function

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2
Q

What is RM?

A

Any therapy that induced regeneration of tissue or organs following disease or injury.

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3
Q

How can RM be achieved?

A

Through gene/cell therapy or pharmaceuticals.

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4
Q

Name the factors in the TE triad.

A

Cells
Scaffolds
Regulators

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5
Q

What is the criteria for TE designs?

6

A
  1. Disease/Stage-specific
  2. Clinically applicable
  3. Achieve function
  4. Mimic native tissue
  5. Host response
  6. Shelf life/storage
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6
Q

What are the goals of TE?

4

A
  1. Decrease patient symptoms
  2. Decrease disease progression
  3. Increase function
  4. Mimic native tissue
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7
Q

Name the 3 critical steps in TE.

A
  1. Define your problem
  2. Develop design criteria
  3. Test appropriate models to mimic the clinical situation
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8
Q

Name the different cell sources.

3

A

Autologous
Allogenic
Xenogenic

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9
Q

What are autologous cells?

A

Patient’s own cells

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10
Q

What are allogenic cells?

A

Human cells. Another person’s cells

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11
Q

What are xenogenic cells?

A

Cells taken from different species

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12
Q

What are the 3 characteristics of autogenic cells?

A

Not rejected
Require isolations + expansion
Limits in cell type

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13
Q

What are the 3 characteristics of allogeneic cells?

A

Can be off-the-shelf
Often require immune suppression
Concerns about disease transmission

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14
Q

What are the 3 characteristics of xenogenic cells?

A

Require immune suppression
Concerns about animal-human viral transmission

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15
Q

What are stem cells?

A

Cells that can divide indefinitely in culture and can give rise to specialized cells

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16
Q

Why are stem cells important in TE?

A

They can regenerate and repair tissues.
Can secrete proliferative and anti-inflammatory growth factors

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17
Q

What are the 3 types of stem cells?
Describe each one.

A

Totipotent: can give rise to any cell type + embryonic tissue
Pluripotent: capable of giving rise to most tissue
Multipotent: gives rise to many cell types

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18
Q

What are the 4 different stem cell sources?

A

Adult - multipotent
Fetal - pluripotent/multipotent
Embryonic - pluripotent
IPSCs - pluripotent

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19
Q

Name the 3 characteristics of adult stem cells.

A
  1. Small numbers
  2. Isolation and expansion isn’t well-defined
  3. Limited ability to expand in culture
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20
Q

Name the 3 characteristics of embryonic cells.

A
  1. Highest degree of pluripotency
  2. Isolated from embryos
  3. Ethical concerns
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21
Q

Name a characteristic of IPSCs.

A

Converted from fully differentiated somatic cells into ESC-like cells

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22
Q

How do we induce differentiation?

6

A

Soluble factors
Cell-cell interactions
Matrix interactions
Cell shape and polarity
Dynamic stress
Oxygen tension

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23
Q

What are the 2 types of cell-cell interactions?

A

Homotypic: contact-mediated adhesion molecules
Heterotypic: soluble paracrine factors

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24
Q

Why are scaffolds important in TE?

4

A
  1. Serve as a matrix for cell adhesion to facilitate/regulate cell processes
  2. Maintain shape + structure
  3. Cell delivery vehicles
  4. Serve as barrier
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25
Q

What is the ECM composed of?

6

A
  1. Insoluble macromolecules
  2. Collagens
  3. Elastin
  4. Proteoglycans and glycosaminoglycans
  5. Cell adhesion molecules
  6. Water (67%)
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26
Q

Name the 4 ECM characteristics.

A
  1. Determine shape and maintain homeostasis
  2. Maintain structure and chemical makeup
  3. Not static
  4. Changes during development
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27
Q

What are polymers?

A

Long-chained insoluble macromolecules made of repetitive units

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28
Q

What are hydrogels?

A

Water-swollen, crosslinked, insoluble polymers

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29
Q

Name 3 natural polymers.

A

Agarose
Collagen
Hyaluronic acid

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30
Q

Name 3 synthetic polymers

A

PEG
PMMA
PGA

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31
Q

How do cells adhere?

3

A
  1. Protein absorption
  2. Modifications to cell adhesion ligands
  3. Results from bioactive surfaces that mediate cell attachments
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32
Q

What should be taken into consideration when designing a scaffold?

4

A
  1. Chemical composition
  2. Structure
  3. Degradation rate
  4. Mechanical properties
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33
Q

Where is integrin binding involved?

6

A
  1. Cell adhesion
  2. Migration
  3. Survival
  4. Growth
  5. Differentiation
  6. Gene expressions
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34
Q

Why is chemical composition important in scaffolds?

A

They can regulate cell function through chemical makeup

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35
Q

What 2 scaffold design characteristics are dependent on chemical makeup?

A

Degradation rate and mechanical properties

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36
Q

What changes can occur in the degradation rate?

A

Too rapid = no proper regeneration
Too slow = interferes with remodeling

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37
Q

What are the steps in 3D bioprinting?

A
  1. Imaging
  2. Design
  3. Material
  4. Cell type
  5. Bioprinting
  6. Application
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38
Q

Name the 3 types of bioprinting.

A
  1. Inkjet - droplets
  2. Microextrusion - continuous
  3. Laser-assisted - drop-cell
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39
Q

What is electrospinning?

A

An electric field draws a polymer stream out of the solution.

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40
Q

What is phase separation?

A

Separation of polymer solution into polymer-rich/poor layers

41
Q

Name 3 pros of phase separation.

A

Similar to setting gel
Control macroporous architecture
Consistent

42
Q

Name 2 cons of phase separation.

A

Limited to several polymers
Small production scale.

43
Q

What are nanofibers?

A

Spontaneous organization into stable structure without covalent bonds

44
Q

Name 2 pros of nanofibers

A

Injectable
Can achieve a small diameter

45
Q

Name 2 cons of nanofibers.

A

Limited to several polymers and hydrophobic/philic interactions
Small size

46
Q

What are 2 types of design modifications?

A

Chemical and Biological

47
Q

Describe the two types of biological modifications.

A
  1. Noncovalent: Adherance through hydrophobic and electrostatic interactions
  2. Covalent: Presence of reactive group on the surface (OH, NH2, SH, etc.)
48
Q

What are the 2 degradation factors in biomaterials?

A

Enzymatic degradation and hydrolysis

49
Q

Name the 2 enzymatic degradation characteristics.

A
  1. Amount and types of enzymes at implantation sites
  2. # of cleavable moieties in the polymer
50
Q

Name 4 characteristics of hydrolysis.

A
  1. Chemical reactivity
  2. Presence of primary and secondary bonds
  3. Media amount
  4. Surface area
51
Q

Through what can scaffolds be sterilized?

A

Type of sterilization and time taken

52
Q

Why is time important while sterilizing?

2

A

It reduced infection
Removes/kills microorganisms

53
Q

What are the 3 types of sterilization methods?

A

Steam
Ethylene oxide
Radiation

54
Q

What are the characteristics of steam sterilization?

3

A
  • Autoclaving
  • Denatures proteins and lipids
  • Effective, quick, simple, no toxicity
55
Q

What are the characteristics of ethylene oxide sterilization?

3

A
  • Toxic, flammable
  • Chemical alteration of nucleic acids
  • Need to purge to allow aeration
56
Q

What are the characteristics of radiation sterilization?

2

A
  • Done through Gamma rays or electron beam
  • Ionizes important cellular elements, including nucleic acids
57
Q

What are cytokines?

A

Polypeptides (proteins) that regulate cell function

58
Q

What are growth factors?

A

They stimulate or inhibit several different cell functions.

59
Q

What do growth factors inhibit?

6

A
  • Cell proliferation
  • Differentiation
  • Migration
  • Adhesion
  • Gene expression
  • Secretion
60
Q

Name 3 characteristics of protein delivery.

A

Expensive
Short half-life
Diffuse from the target site

61
Q

Name the 3 factors needed for gene delivery.

A
  1. Expression casette
  2. Non-viral vectors
  3. Viral vectors
62
Q

What are the three types of controlled release?

A
  • In scaffold
  • Linked to scaffold
  • Transport transfected cells
63
Q

What cells are mostly used for GFs?

A

Fibroblasts

64
Q

What are the three types of mechanical regulators?

A

Flow - endothelial
Mechanical - muscle
Electrical - neurons

65
Q

Name 6 characteristics of innate immunity.

A
  • Non-specific
  • From birth
  • Anatomic barriers (skin / mucous)
  • Physiological barriers (body temp)
  • Phagocytic cells
  • Inflammation
66
Q

Name 3 characteristics of acquired immunity.

A
  • Specific
  • Lymphocyte activation
  • Vaccines
67
Q

What is inflammation?

A

Non-specific response to damaged tissue

68
Q

Signs of acute inflammation

A
  • Redness
  • Swelling
  • Pain
  • Heat
69
Q

What are the first cells in the site of inflammation?

A

Neutrophils

70
Q

What are 4 characteristics of neutrophils at the site of inflammation?

A
  • Stick to capillary
  • Penetrate between endothelial cells
  • Move to surrounding tissue
  • Emigrate in min/hrs
  • Activate when engaged with foreign particles
71
Q

What cells substitute neutrophils at the injury site?

A

Monocytes

72
Q

Why do monocytes replace neutrophils?

A

Neutrophils are short-lived (24-48 hrs)
Chemotactic factors for neutrophil emigration are deactivated in later stages.

73
Q

Why are macrophages important?

A

They are phagocytic
They release cytokines
They produce a large # of biologically active products

74
Q

What are the characteristics of chronic inflammation?

A
  • Persistence of macrophages, monocytes, and lymphocytes.
  • Proliferation of blood vessels and connective tissue
  • No exudates
75
Q

What GFs are present in inflammation?

A

PDF
FGF
TGFB
IL-1
TNF

76
Q

What is the end stage of healing called?

A

Fibrosis

77
Q

When does resolution happen?

A

When no biological changes occur

78
Q

What are the steps of resolution?

A
  1. Extrusion
  2. Resorption
  3. Integration
  4. Encapsulation
79
Q

What is histology?

A

The study of tissue’s structure and arrangement

80
Q

What does hematoxylin and eosin (H&E) staining represent?

A

Hematoxylin: basophilic structures, blue-purple staining, nucleic acids
Eosin: eosinophilic structures, bright pink staining, cytoplasm

81
Q

Name 5 pros of natural materials for scaffolds.

A
  • Lower risk of immune rejection
  • Promotes structure remodeling
  • Biodegradable
  • Inherit structural resemblance
  • Interaction with surrounding env.
82
Q

Name 5 cons for natural materials for scaffolds.

A
  • Hard to control degradation properties
  • Storage –> expensive
  • Cross-contamination
  • Hard to manufacture in bulk
  • Less acceptable to structural and mechanical properties
83
Q

Name 5 pros for synthetic materials for scaffolds.

A
  • Uniform composition
  • Controllable and reproducible
  • Can be optimized
  • Ethical
  • Engineered with desired properties
84
Q

Name 5 cons for synthetic materials for scaffolds.

A
  • Binding sites must be incorporated
  • Poor biocompatibility
  • Higher in cost production
  • Risk of rejection
  • Toxic byproducts
85
Q

Name 5 pros of ESCs

A
  • Longterm expansion
  • Disease modeling
  • IVF Increased
  • Reprogrammable
  • Low genetic manipulation
86
Q

Name 5 cons for ESCs

A
  • Ethical concerns
  • Unclear history and characteristics
  • Heterogeneity
  • Genome editing is slow
  • High risk of tumorigenesis
87
Q

Name 5 pros of ASCs

A
  • Less prone to genetic modification
  • High regenerative ability
  • No immune response (autologous)
  • Easy to culture
  • Low process costs
88
Q

Name 5 cons of ASCs

A
  • Limited differentiation potential
  • Age donors can affect
  • In vivo/In vitro differences
  • Reprogram takes several weeks
  • Poorly characterized
89
Q

Name 5 pros of IPSCs

A
  • Low immunogenicity
  • Disease modeling
  • Low ethical concerns
  • Easy to harvest
  • Has reached clinical trials
90
Q

Name 5 cons of IPSCs

A
  • Can form teratomas
  • Immune response
  • Epigenetic memory
  • Genomic instability
  • Expensive
91
Q

What is the consumption rate of oxygen?

A

0.05 - 0.5 umol
10^6 cells/hr

92
Q

What is the consumption rate of glucose?

A

0.2 umol
10^6 cells/hr

93
Q

What is the consumption rate of amino acids?

A

0.1 - 0.5 umol
10^9 cells/hr

94
Q

What is the equation to determine supply?

A

S = kpx^2 / DCo

95
Q

Give 3 characteristics of a rapid regulatory pathway.

A
  1. $0.2-2 million in costs
  2. Quality, steriability
  3. 1-2 years
96
Q

Give 3 characteristics of the short-term regulatory pathway.

A
  1. $2-20 million in costs
  2. Safety, 501K
  3. 1-3 years
97
Q

Give 4 characteristics of medium-term regulatory pathways.

A
  1. $30-200 million in costs
  2. Animal/safety studies, IDE, PMA
  3. Clinical trials
  4. 1-6 years
98
Q

Give 4 characteristics of long-term regulatory pathways.

A
  1. $50- >300 million dollars
  2. Animal/safety studies, IND, NDA
  3. Clinical trials
  4. 1-8 years

Tissue Eng (1 decks)

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