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BioMaterials (Tresco) > Exam 2 > Flashcards

Exam 2 Flashcards

1
Q

With regards to the FBR, how does the acute phase of inflammation differ from the chronic phase?

A

Initiated by injury to blood vessels
Self assembly of fibrinogen, forms provisional matrix. allows cells into damaged area.
protein adsorption
complement activation
macrophage activation and WBC recruitment and Neutrophil!!
-most abundant WBC, phagocytic, chemotactic. unclear what happens to them, only beginning to understand role in FBR

Neutrophils are the biggest difference
activated macrophages never disappear
no resolution of wound

FROM REVIEW:
chronic involves persistant mac. recruitmant, granulation tissue formation, fibrous encapsulation

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2
Q

How does complement sustain inflammation

A

Answer from review:
C3A, C5A proteins diffuse away from gradient and macrophages are activated (complement sustains FBR) which cause leaky vessels.

Macrophages come down to surface and phagocytize things on surface. Take proteins to present to b-cells. White blood cell trafficking. Cause leaky vessels. which allows more macrophages to continue to bind.

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3
Q

What are the general functions of macrophages in the FBR?

A

To kill, cleanup and tissue remodeling

eliminate dust, allergens, and microorganisms.

clearance of pathogens and toxins

bone resorption

recognition and removal of enteric pathogens, tolerance to food antigens and microbiota

antigen capture and presentation to B cells

phagocytosis and antigen presentation

secrete molecules that recruit other macrophages, result in the proliferation of endothelial cells and fibroblasts - they drive the foreign body response

  • the more activated the greater the foreign body response

phagocytosis, recruitment of other macrophages, stimulation of fibroblasts and endothelial cells, stimulate ECM production

Phagocytic, recognize bound antibodies and complement coated foreign bodies.

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4
Q

Where do macrophages come from?

A

all macrophages have early origins. one is from yoke sack (stem cells), the second from fetal liver.

Also derived from bone marrow and then from monocytes!!!!!, activated monocytes.

All tissues have resident macrophage populations

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5
Q

What are three general secretory products of macrophages

there’s 5…

A

Diverse Secretions
1 toxic oxygen-deprived products (superoxide, H2O2, hydroxy radicals, singlet oxygen)
2 Digestive Enzymes (plasminogen or neutral proteases)
3 Monocyte Chemoattractant Protein (MCP-1)
4 Proinflammatory Cytokines: TNF-alpha, a cytotoxic moleucle. Interleukin-1 beta, stimulates fibroblast prliferation. Transforming growth factor beta (TGF-beta), induces cell death and increases…. (follow up)

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6
Q

How do fibroblasts participate in the FBR

A

Cell type that secretes ECM and secretes angiogenic factors to rebuild tissue

Motile cells that are found in tissues whose primary function is to maintain tissue integrity by responding to injury, inflammation or forces by making ECM

change tissue based on changing loads

involved in chronic phase of FBR

express more actin and myosin-myofibroblastic phenotype
overproduce ECM-responsible for fibrous encapsulation

fibrous encapsulation

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7
Q

What roles does collagen play in the FBR?

A

structural components of all connective tissues natural hemostatic and regenerative biomaterial

Collagen-structural components of all connective tissues natural hemostatic and regenerative biomaterial, cartilage bone tendons ligaments fascia skin

collagen is the molecule that makes up the fibrous capsule around the implant - collagen is an ECM molecule secreted by fibrobalsts in the late stage of the FR which makes up the fibrous capsule

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8
Q

Describe (list) thee different types of devices chronically implanted in the CNS

A 
Recording Brain electrodes, 
--to detect epilipsy
-to map activity
Deep Brain stimulating Electrodes
-Pacemaker like battery, stimilutates certain spots to help release certain neurotransmitters
Aneurysmal Clip
-used to clamp aneurysms in the brain
Sustained Delivery System (wafers)
-Eventually degrade,
-elate chemotherapeutic drugs
Cell Encapsulation device
-diffuses lacking neurotransmitters to the correct spot in the brain.
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9
Q

Describe a typical experimental approach for studying the tissue reaction to a device placed in brain tissue

A 
Prepare Clean and Sterile Implant
Select Target tissue
Select Animal Model (Select the N)
Set the time points of analysis
Analyze retrieved implant
1. Select Hisotological approach
2. Use Quantification
3. Statistical Analysis
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10
Q

Describe how you would detect and image cells specific to brain tissue using immunohistochemistry.

A

antibodies that targets things only expressed by a specific cell, in this case the brain

GFAP: glial fibrillary acidic protein - Astrocyte Biomarker (Unique to brain)

ED-1 (CD68): Iysosomal protein-Activated Macrophage Biomarker

NeuN: neuronal nuclie-Neuron Biomarker

Non-specific biomarkker for cell nuclei
DAPI: 4’,6-diamidino-2-phenylindole
Marks all cell nuclei

Primary IHC:

  • Target a cell-specific marker with an appropriate antibody against that cell-specific marker
  • The antibody has a fluorophore or alternative marker that can be imaged

Secondary or Indirect IHC

  • Target a cell-specific marker with an appropriate primary antibody against that cell-specific marker
  • Target the primary antibody with a secondary antibody that binds the primary
  • The secondary antibody has a fluorophore or alternative marker that can be imaged
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11
Q

Describe how to perform indirect immunohistochemistry

A

Uses antibodies
Primary antibody recognizes target molecule
bind to the surface
buffer washes away all unused primary antibodies
secondary antibodies are labeled with a color or flourescent marker.
Secondary antibodies interact and bind w/ primary antibodies
Using 2ary antibodies, the signal is magnified

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12
Q

Describe the three sectioning planes used in histology

A

follow up. check w/ Tresco

Coronal-deli meat slices
Horizontal- cutting along the length of the brain
Sagittal-cuting along length and into plane of the board

CORONAL PLANE:
Vertical sectioning, dividing object into posterior and anterior portions
The SAGITTAL PLANE:
Vertical sectioning, dividing object into left and right
HORIZONTAL PLANE
Horizontal sectioning dividing object into top and bottom

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13
Q

Describe the general features of the foreign body response to an implanted electrode in rat brain

!!!

A
  • Activated Macrophages @ interface both on device and tissue (inflammation) (CD68)
  • Leaky blood brain barrier
  • hypotrophic response in GFAP protein-fibrous encapsulation.
  • NeuN indicates loss of neurons, which are never regenerated near FBR

Shape of the FBR Mirrors Implant Geometry

Activated macrophages @ abiotic/biotic surface drives FBR
activated macrophages secrete TNF-alpha, ROI’s, Proteases, Growth Factors, IL 1-beta, MCP-1 which do not diffuse easily through brain tissue.

24 hours after implantation, brain shows sign of hemorrhage

Tissue loss associated w/ array implantation likely caused by vascular damage

REVIEW: IgG-shows leaky vessels

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14
Q

Provide a plausible hypothesis for why the FBR is very localized at the interface of an implanted electrode;

A

Activated macrophages occupy the biotic/abiotic interface and behave as a delivery system and modulate tissue remodeling. Macrophages secrete a lot of things (cytokines), and they match the geometry of the implant, which means the FBR is also centered around implant.

little extracellular space so mac. secretions don’t diffuse very far

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15
Q

Why do you think high density penetrating electrode arrays, like the UEA, create lesions in rat cortex following implantation?;

A

The brain is highly vascularized which makes it extremely unlikey that the UEA could be inserted without vascular damage.

cannot put one of these in without damaging blood vessels which leads to large amount of hemorrhaging which then causes the tissue loss

REVIEW: damaged vasculature

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16
Q

Why does the UEA behave like a sustained release device for proinflammatory cytokines?;

A

With macrophages dominating the surface interface, there are proinflamatory cytokines that are constantly being released near or around the implant.

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17
Q

How would you redesign the UEA to reduce the impact of the FBR? Justify your design solution.

A

This is an opinion, think about it critically. and answer

limit surface areas, , coat w/ anti inflammatory stuff, reduce the number of spikes, just have same multiple measuring points on fewer spikes.

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18
Q

With regard to this course, what is meant by the term “healthcare-associated infections? and provide a synonm

A

Nosocomial Infections

-Infections that are acquired while receiving medical treatment in a healthcare facility
-from ‘nosos’ greek for disease
- results from introduction of bacteria during device insertion or an implant procedure or from attachment of organisms to an indwelling device, and subsequent proliferation on its surface.
-most are bacterial infections
- good clinical practice (aseptic or sterile technique) reduces but does not eliminate the occurrence of infection.
-

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19
Q

List three common bacteria found on the surface of the human body

A

staphylococcus epidermidis

staphylococcus aureus

streptococcus mitis

streptococcus salivarius

streptococcus mutans

enterococcus faecalis

streptococcus pneumoniae

streptococcus pyogenes

neisseria sp.

neisseria meningitis

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20
Q

Why are humans and their pets sources of bacterial contamination?

A

Skin is primary source of infection. 1.5 g of skin shed per day, which are covered in bacteria. Every human activity leads to skin flaking off, as it flakes off, it spreads bacteria.

animals and even plants shed cells

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21
Q

List 4 general strategies to prevent device related infections

A

Minimize contact- clean room conditions
kill everything in contact-sterilization
minimize binding at contact-surface coating
kill after contact- anti-infective coatings

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22
Q

What air handling filter specification should you use in building a surgical suite or a cell culture laminar flow hood?

A

(Number of particles removed/ cubic foot)

.3 micron air filter
room ought to be ventilated by an efficient bag filter or high efficiency particulate air filter (HEPA) system

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23
Q

What implants design element is likely to result in infection? and why?

A

Breaching of skin barrier. PERCUTANEOUS Anything that allows entrance to the body from the outside. Lack of soft tissue interface. more than 2 weeks = 50% infection

when chemotheruaputic drugs reduce the production of immune system.

24
Q

What are the three most commonly infected biomedical devices?

A
  • Foley Catheter-associated urinary tract infections
  • Central Venous Catheter-assosicated bloodstream infection (High chance of morbidity)
  • Ventilator Tube- associated pneumonia

most infections are extralumenal (rarely is the source of infection the infusate)

25
Q

Using a foley catheter as an example explain why it gets infected if it remains in place for several weeks?

A

Skin particles flake off, and the bacteria is flaking off under the gown/covers. Bacteria flakes onto tube and eventually gains access to the body.

(MORE TRUE) Most infections come from bacteria from the GI tract. Patients excrete, and it’s spread over skin near butt/catheter. Bacteria gains access to body by catheter (on exterior). Bacteria in fluid swim upstream. form biofilm, can clog catheter
-Bug (E. Faecalis) crawls up, uses proteases to break down epithelial cells, enters blood streams

26
Q

Why is CLABSI a more serious infection than CAUTI

A

CAUTI=(Catheter-associated Urinary Tract Infections)
CLABSI=(Central LIne-associated Blood Stream Infections) -major cause of morbidity and mortality in US
-Bacteria are from upper torso of body

CLABSI causes Sepsis- proinflammatory cytokines are released to all of the organs in the body. has access to blood stream and infects everywhere

27
Q

What is meant by the term biofilm? and why are they especially problematic?

A

Polysaccharide secretion of bacteria (ECM)-negatively charged
Biofilms form on implanted devices. Ca2+ cross links and forms a barrier which excludes antibodies, complement cells and antibiotics. Once biofilm forms, an implant must be removed.

28
Q

What is meant by the term sterilization? List the two most common commercial sterilization processes and their killing mechanism?

A

Def: a process used to render a product free from VIABLE microorganisms (absence of living organisms)

Sterility Assurance Level (SAL): Probablility of a vaiable microorganism being present on the product unit after sterilization and generally accepted max 10-6 (one in one million is nonsterile)

Dry heat ( uses destructive oxidation)
Steam(autoclaving) (pressurized steam)
Ethylene Oxide (1)
Radiation (2)-gamma ray

look up killing mechanism

29
Q

Why is it important a clean a device before sterilization?

!!!

A

Even dead bacteria can illicit the FBR, The mere presence of biological material will lead to the FBR.

Endotoxins (part of bacterial cell wall (graham negative) aka liposaccharides). activate macrophages and cause inflammation–even dead bacterial will increase FBR

sterilization doesn’t get rid of surface ENDOTOXINS

30
Q

Explain what is meant by the term “surface coating technology”

A

Surface coating technology is a coating applied to biomedical devices designed to protect and enhance the surface functionality. It is important to the biomedical industry because it will impact the functionality of your biomedical device and can be tailored to fit needs of the device.

engineering the surfaces of devices to improve function of device. Alter surface properties (chemistry of a device)

31
Q

Why is surface coating technology preferred in the biomedical device industry?

A
  • modify and improve existing device
  • keep majority of manufacturing processes and facilities in place
  • piggyback on previous regulatory approvals

Surface coating enhances to safety and efficacy of the device without completely changing the device. These allow companies to modify and improve existing devices, keep a majority of current bulk manufacturing processes & facilities and piggyback on previous on previous regulatory approvals.

32
Q

List the characteristics of an ideal surface coating?

A

Improves device biocompatiabliltiy
reduces adhesion of proteins, cells or microbes
durable/stable
uniformly covers the surface
is compatible w/ standard sterilization techniques
easy to apply
does not add excessive costs to the manufacturing process
does not raise additional regulatory issues

33
Q

Describe two different methods used to influence protein binding at interfaces and provide a rationale for how each of them is believed to function?

A 
Gas Plasma (vacuum) or Corona Treatment(atmospheric)
-

Surfactant Adsorption

Electrostatic Based, Layer by Layer deposition

  • adding positive and negative charges to the starting substrate
  • eventually diffuse through material

Covalent Immobilization

34
Q

What is meant by the term plasma treatment? How does it work?

A

Surface treatment made using an ionized gas such as Ar, O2,

Gas runs through cathode/anode becomes energized (may add particulates) and added to a coating)

so reactive they don’t remain on the surface for very long. Can add oxygen to any surface

Plasma treatment is a surface treatment made using a gas ionized under a vacuum and is used to remove contaminates, temporarily energize the surface, and add new molecular groups to the surface.

REVIEW: difference between plasma and corona. PLASMA USES VACUUM

35
Q

What is meant by the term “adsorption or surfactant based coatings? How is it used as surface modifying agent?

A
  • uses Amphiphilic molecules
  • -Hydrophobic region adsorbs to hydrophobic biomaterial
  • -hydrophilic region orients toward water.

Add these surfactants to the surface of biomaterial, ideally they’re covalently bonded to change surface chemistry. Permanently effects surface chemistry

36
Q

What is an application weakness of adsorption or surfactant based surface treatments?

A

If coatings aren’t covalently bonded to device, the coating may react with things in the body, or diffuse from the surface of the device into the body. Not great long term. can be replaced by other adsorbant species.

37
Q

What is meant by the term covalent surface immobilization and what are its advantages over surfactant based techniques?

A

Covalent surface immobilization refers to a method used to coat a surface by using covalent bonds. The primary advantage over surfactant based techniques is the in the bonding strength of the covalent bond which far exceeds that of surfactants which are linked primarily via weaker hydrophobic interactions. Advantage #2 is that the immune response is not as great

Covalent surface immobilization is a method used to layer a surface with a new molecule covalently bonded to it. It’s advantages over surfactant based techniques include a covalently bound molecule, which proves to be much stronger than surface adhesion.

38
Q

What is meant by the term steric repulsion?

A

explained with polyehtylene oxide - steric hindrance - long chain covered with polyethylene oxide, cannot absorb so there is repulsion, physically exclude something from its space because it moves or is there - glycocalyx- polysaccahride chains covered in water that as the move around they create a space that proteins cant enter- long chain hydrophilic molecules like PEO polymer chains adsorbed or covalently attached can move at elevated temperatures and exclude proteins from interacting with the more hydrophobic biomaterials.

This occurs when the size of groups within a molecule prevent chemical reactions that would be able to occur in smaller molecules. Bringing together atoms has an energy cost as the electron clouds overlap.

Steric repulsion is the resistance to protein adsorption using steric hindrance of bound surface molecules.

REVIEW: physical barrier, molecule takes up space which prevents things from adsorbing. the molecules move and creates a wider steric repulsion zone. Focus on dynamic movement of molecules on surface
ex. PEO-binds water. surface coating technology

39
Q

What is PEO and how is it employed in surface coating technology?

A

Polyethylene Oxide. O along polymer backbone allows H-bonding w/ H2O. Works best in fluid filled tissues (blood stream)
uses:
-increase solubility of hydrophobic molecules
-Lubricant
-Adhesion resistant coating.

repulses things sterically

40
Q

Explain what is meant by the term “site specific delivery”? Why is it advantageous in the context of an antimicrobial coating?

A

Definition-Locating active agents or drugs only at the surface of or in the vicinity of the device;

Preferable to administering the same drugs systemically;

Local administration from the device surface concentrates the drug at the precise site where it is needed;

Decreases potential for bacterial resistance;

Avoids systemic side effects.

Chronic Dosing is the Limitation with Site Specific Drug Delivery

Effective delivery
Must deliver appropriate agents to kill off the specific pathogens that infect different devices;
Sufficient amounts of the agent released from the device need to rise to a therapeutic level;
Duration of release must be appropriate for the condition.

Site specific delivery is the act of locating active agents or drugs only at the surface of or in the vicinity of the device. It is advantageous to antimicrobial coating because it administers drugs in a systematic fashion, decreasing the potential for bacteria resistance.
Decreases bacterial resistance, avoids systemic side effects

41
Q

Within the context of biomaterials, what is meant by the term hollow fiber?

A

Semipermeable tubular materials made of synthetic or natural materials that are capable of conducting flow in an axial direction or providing a conduit to guide tissue regeneration or isolate cells.

semipermeable: cabable of separating things

42
Q

List several important biomedicl applications of hollow fiber membranes?

A 
Liquie sterilization -cell culture .4 microns
Excretory assist devices
-dialysis
-liver assist devices
-blood gas exchangers
Implantable biosensors
vascular grapfts
nerve repair
43
Q

Describe the working principle behind hemodialysis?

!!!

A

Process of removing toxic molecules from blood (uremic toxins)
3 sessions of 4 hours/week

Blood flows in to dialyzer and is cleaned using the process of diffusion and ultrafiltration
-extracellular fluid=dialysate
(probably should follow up)

REVIEW image: Describe biomaterial/working principle:
blood comes from artery-pumped through dialysis membrane-dialysate pumped in the other direction. filtration by diffusion.
lots of blood contact through tubing.

44
Q

How is membrane permeability determined?

A

Determined by pressurized flow filtration assays. Using fluorescently-labeled dextrans, performance is determied due to their low non-specific binding to mebrane of different molecular weights.

Molecular weight cut off (MWCO): MW @ which 90% are retained by membrane

Sieving assay’s

sieving curve!! graph..

45
Q

What is meant by the term phase inversion?

A

Controlled precipitation: the proces used to form hollow fiber membranes.
wall is porous, structure traversed by an interpenetrating pore network or channels across the wall structure
-densest part of the wall is called the skin.

46
Q

Generally speaking, how are hollow fibers made?

A

thermoplastic
-most common: PS, PAN,

process: Dry, jet wet spinning, a phase inversion technique

polymer taken and put into solution. Ran through spineret, creates hollow fiber

air gap

solvent=DMSO
nonsolvent=water

(check slides for image)

REVIEW
as soon as solvent/nonsolvent make contact at spout of spinneret they begin to form

pump of solution and nonsolvent meet in spineret. nonsolvent inside.

47
Q

What types of equipment and reagents are required for fabricating a semipermeable hollow fiber?

A

A polymer w/ sufficient MW and solution density (15-25% w/v): so there is enough chain length to provide chain entanglement after precipitation

  • A solvent for the polymer
  • A miscible non-solvent

spinneret
tubular form is created w/ a spinneret.

48
Q

Provide a definition for the term “elastomer” and provide two examples of the most commonly used materials in this class.;

A

A natural or synthetic polymers that returns rapidly to approximately its initial dimensions and shape after substantial deformation and release of its stress

Real 1: polyurethane
Real 2: Silicone

also: natural rubber(latex)
Elastin

latex  exam gloves
  medical tubing
grafts, pacemakers,
balloon catheters,
joint sockets for prosthetics
heart assist devices
condoms
wound care products
49
Q

Explain why isocyanates are very reactive.

A

def: a functional group that contains 1 N, 1 C, 1 O.
because they’re so electronegative, the have a very reactive resonance structure. highly reactive, reacts with nucleophilic amines and …? (nucleophilic addition)

50
Q

Outline the basic reactions that underlie the formation of a polyurethane oligomer (synthesis);

!!!!!!

A

Synthesized as thermosetting polymers
copolymers that exhibit elasticity

Diol+2 Diisocynate = prepolymer
prepolymer+chain extender=copolymer

Diol:
MW betwen 1000-2000 Da,
2 types used, polyester based (susceptible to hydrolytic degradation) or polyether based

chain extenders:
add amorphous region called soft segment
glycol, gloycol, butanediol, glycerol

51
Q

Provide a structural model that explains the structure/ properties relationship of a typical segmented thermoplastic polyurethane

A

Polyeurethane linkage is also susceptible to hydrolysis. Not cleaved due to aromatic components which are hydrophobic. they resist the attack.

monomer diol–diisocyanate – monomer diol
|||| |||||||
hard segment ————————-soft segment
||||||||||
randomly segmented copolymer

Soft Segment-free rotation increases elasticity and the amorphous microstructure
Hard Segment-aromatic pi bonding increases crystallinity and hardness

52
Q

Explain why some thermoplastic polyurethanes are susceptible to degradation in the bodily tissues.

A

Urethane linkages are susceptable to degradation

53
Q

In the context of this course, define what is meant by the term “silicone” and provide an example of one?

A

Technical name is polysiloxane-incorrectly labeled in 20’s.

R-Si-R
|
O

definition: an inorganic compound with repeating Si-O linkages in the backbone where silicon is also bonde to organic groups.

Linear silicones are fluids at room temp.
mostly thermosetting elastomers

R=Methyl group

PDMS !!!

54
Q

Describe in general terms a synthetic scheme for a silicone and explain why they are difficult to reprocess.

A 
Step 1:
Polymerization
-platinum-based addition reactions (free radical)
--either--
-Condensation reactions

Step 2: Cross-Linking
-no crystalinity-behaves like carbon-behaves like large saturated hydrocarbon chain-amorphous regions of polymer cross linked at random

Thermosets cannot be re-melted or reformed because they’re cross linked. like thermoplastics and must be mixed from a two part liquid silicone that contains cross linker and catalyst

Due to second step.

55
Q

Provide three examples of biomedical devices made from poly(siloxanes).

A

Joint replacement

Cosmetic/Plastic

  • breast implants
  • lap bands
  • testicular prosthectis
  • butt implants

Lubricants for biomedical devices
hyodermic needles

Medical device encapsulation or used in mold making
-pace maker

Condoms

tubing, check valves, gaskets, respirator mask,

-Hand joint replacement

56
Q

List three attributes of poly(siloxanes) that make them a favorite material for biomedical devices.

A

Extremely soft and pliable, easily conform to different cavity shapes;
High gas permeability;
Low reactivity with tissue and body fluids;
Good tensile strength and tear resistance (generally lower than polyurethanes);
Susceptible to degradation in acidic conditions (i.e. stomach);
Hydrophobic (protein adsorption);
Easily sterilized under all conditions.

57
Q

How does a central venous catheter differ from a peripherally inserted central catheter or PICC?

A

def of catheter: tubes that can be inserted into a body cavity, duct, or vessel that allows drainage, administration….

two most common complications are infection/thrombosis

PICCS:
centrally placed, tip ends up in Superior Vena Cava
Peripherally inserted meant it goes into your body at your elbow and threaded into vein
May have a valve at the tip, preventing blood from backing up into the catheter, so heparin is not necessary
can do most normal activities, expcept swimming, or other extreme arm movements

difference:
length and where tip ends up!

BioMaterials (Tresco) (3 decks)

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