Quiz 4: Anti-infective Coating & Infection Prevention Flashcards

1
Q

high infection rate from devices is due to

A

direct exposure to bodily fluids and invasive placement within the body

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2
Q

control of biofilm formation: Antimicrobial approaches

A

Most biomaterials in this category either display or release bioactive molecules, while others promote cell lysis by exposing functional groups that effectively interact with bacteria cell membranes. (Intrinsically bioactive biomaterials: Cationic materials)

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3
Q

Biomaterials releasing bioactive molecules

A

Surface modification with antibacterial agents:
- Antibiotics
- Metal ions
- Nitric oxide

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4
Q

Different strategies to apply bioactive molecules to biomaterial surface:

A
  • Physical adsorption (simplest strategy: surface)
  • Impregnation inside the biomaterial (bulk, e.g., pores)
  • Complexation
  • Conjugation
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5
Q

Antibiotics

A

Examples in orthopedic applications:
- Gentamicin (most common)
- Gentamicin + Vancomycin

Antibiotics-loaded (A-L) bone cement (bone cement impregnation)

Coating implant surfaces with biodegradable materials that release antibiotics in a controlled fashion.
- PLGA or PLA polymer as coatings for Titanium (Ti) implants to deliver gentamicin.
- Mesoporous hydroxyapatite, combined with antibiotics as coatings for dental and bone implants.

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6
Q

Silver

A

Silver (Ag+) accumulates at the cellular membrane and leads to membrane perforation. Also, Ag+ promotes the generation of reactive oxygen species.
- Formulations include Ag salts, Ag oxide, metallic Ag, Ag alloy, or Ag nanoparticles (the most active form).
- Silver has been loaded in PLGA coating for Ti implants; commonly used in urinary catheters; successfully used in wound-dressing materials.

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7
Q

Low-dose Nitric Oxide (NO)

A

kill bacteria primarily due to its reactive and toxic effects on microbial cells.
- Generation of Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS) that are highly toxic to bacterial cells, damaging essential cellular components like DNA, proteins, and lipids.
- Inhibition of enzymes necessary for bacterial cell wall synthesis, susceptible to lysis.
- NO has been shown to be effective in breaking down biofilms— penetrate biofilms and weaken the bacterial community, making individual bacteria more susceptible to immune cells and treatments.

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8
Q

Intrinsically bioactive biomaterials: Cationic materials

A

Promote cell lysis by exposing functional groups that effectively interact with bacteria cell membranes

Cationic polymers with a net positive charge (backbone or side chain) bind to the negatively charged components of the microbial cell membrane promoting membrane disassembly and leakage or lysis.
- chitosan (in wound dressings, drug formulations, and tissue engineering)
- Antimicrobial peptides (Small cationic peptides, 10-50 amino acids, produced by the innate immune system)
- Synthetic cationic polymers (e.g., Polyethyleneimine (PEI), but has cytotoxicity concerns in human cells)

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9
Q

Intrinsically bioactive biomaterials: Cationic materials advantages

A

long lifetime and low propensity to induce antibiotic resistance.

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10
Q

control of biofilm formation: Antifouling approaches

A

Repel microbes, thereby impairing bacterial adhesion and biofilm formation.
(1) Hydrophilic polymers
a) PEG
b) Zwitterionic materials

(2) Superhydrophobic materials
a) “Lotus leaf effect”, unique self-cleaning and bacterial-fouling resistant properties.
b) Can be obtained by combining hydrophobic moieties with proper micro/nanoscale topography.

(3) Materials with nano/microscale surface texture
Bactericidal surfaces: disrupt bacterial cell, causing death (e.g., dragonfly and cicada wings and geiko skin)

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11
Q

Materials with nano/microscale surface texture: nanopillars or nanospikes

A

Bactericidal Mechanism of nano-
textured surfaces:
Bacteria cell walls stretch and disfigure when they interact with textured surfaces. Stretching occurs in the regions between structures, and if sufficient, cell rupture and death occur.

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12
Q

Quorum sensing

A

bacterial cell-cell communication within biofilms, primarily known
as “quorum sensing,” is facilitated by the secretion of signaling molecules called autoinducers, which build up concentration gradients within the biofilm environment, allowing bacteria to coordinate their behavior based on the local population density.

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13
Q

control of biofilm formation: Biomaterials affecting biofilm
architecture

A

Biomaterials expose enzymes or quorum sensing (QS) molecules that modify biofilm structure and enhance biofilm removal by either physical methods or antibiotic treatments.

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14
Q

Biomaterials modified with QS-quenching enzymes

A
  • QS signal molecules include lipids and cyclic or linear peptides.
  • Enzymes that degrade these molecules can lead to reduced biofilm thickness, reduced biofilm formation, and interfere with QS pathways
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15
Q

Biofilm matrix-degrading enzymes

A
  • Dispersin-B and DNase I
  • These two enzymes can degrade components in the biofilm EPS matrix
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16
Q

Infection Prevention: general principles of sterilization or disinfection of medical devices

A

In general, reusable medical devices or patient-care equipment that enters normally sterile tissue or the vascular system or through which blood flows should be sterilized before each use.

Sterilization means the use of a physical or chemical procedure to destroy all microbial life, including highly resistant bacterial endospores.

The major sterilizing agents used in hospitals are a) dry heat, b) moist heat by steam autoclaving, c) ethylene oxide gas, and, d) radiation

Disinfection means the use of a chemical procedure that eliminates virtually all recognized pathogenic microorganisms but not necessarily all microbial forms (e.g., bacterial endospores) on inanimate objects.

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17
Q

which of the following best describes the function of an anti-infective coating on a medical device?
it improves the strength and durability of the device
it prevents the adhesion and growth of microoganisms
it promotes biofilm formation on the device surface
it enhances the device’s compatibility with human tissue

A

It prevents the adhesion and growth of microorganisms

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18
Q

which dressing type prolongs silver release for sustained antimicrobial efficacy?
hydrophilic dressing
hydrophobic dressing
no difference
cannot be determined

A

hydrophobic dressing

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19
Q

which property of a biomaterial is most likely to reduce bacterial biofilm formation on a medical implant?
micro-scale roughless
high hydrophobicity
nanoscale roughness
high porosity

A

nanoscale roughness

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20
Q

why are nanotextured surfaces effective in antifouling applications?
they attract bacterial cells with a positive charge, preventing detachment
they make grooves and pits that trap bacteria, making them adhere strongly
they form a hydration layer that prevents bacterial attachment
they have nanoscale features that disrupt bacterial cell walls on contact

A

they have nanoscale features that disrupt bacterial cell walls on contact

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21
Q

true or false: anti-infective coatings can prevent biofilm formation by directly killing bacteria or blocking attachment to surfaces

A

true

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22
Q

true or false: hydrophilic coatings are generally less effective in reducing bacterial adhesion compared to hydrophobic coating

A

false

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23
Q

sterilization

A

validated process used to render a product free from viable microorganisms
sterile: refers to the absence of microorganisms

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24
Q

sterility assurance level (SAL)

A

probability of a single microorganism being present in a sterilized product. a statistical measurement of the likelihood that a sterilized item is still non-sterile. SAL=10^-6
most processes are designed to overkill sterilization

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25
Q

disinfection

A

reduces the number of pathogens on surfaces. it is usually less effective than sterilization. used for environmental surfaces and non-critical items.

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26
Q

factors affecting the outcome and effectiveness of the sterilization process

A

cleanliness of the material being sterilized
nature and extent of initial bioburden
design, manufacture, and assembly of device/implant
chemistry/reactivity of underlying biomaterials
features such as convoluted channels or internal porosity

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27
Q

sterilization techniques

A

radiation sterilization
ethylene oxide sterilization

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28
Q

thermal sterilization

A

used mostly in hospital settings
paired with established validation processes
often not possible when biologics or liquids are added to device or when a combination product is repacked for use (aseptic processing of the product preferred to ensure SAL can be maintained)

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29
Q

Sterilization methods for liquid products

A

use one of the variations of steam sterilization
small volume parenteral also might be compatible with radiation sterilization
avoid aseptic filtration/fill unless absolutely dictated by product compatibility

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30
Q

Sterilization methods for non-liquid products

A

steam, dry heat, and radiation sterilization are preferred over ethylene oxide
these processes are relatively simple and do not leave toxic residues in the product

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31
Q

Radiation-based techniques: safety concerns

A

possible lethal exposure to radiation and ozone inhalation
safety measures: shielding and robust interlocks implemented in radiation processing as well as ozone monitors and adequate ventilation

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32
Q

Radiation-based techniques used for

A

mass-produced medical devices bc of its simplicity and convenience in terms of large-scale processing, including sutures, gloves, face masks, syringes, surgical stapler etc

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33
Q

Radiation causes

A

DNA damage in bacteria and viruses preventing pathogens from reproducing and inactivating them.

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34
Q

Radiation sterilization occurs through three methods:

A

gamma, electron beam, x-ray

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35
Q

material considerations for radiation sterilization

A

free radicals cause scission to DNA but may also disrupt material properties of polymers resulting in chain scission or cross-linking, or even reacting with oxygen to damage synthetic or biological materials of the sample
material of the product remains inert to the low doses that occur during

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36
Q

material considerations for radiation sterilization: mitigating alteration ex

A

PLGA experiences a decrease in MW after sterilization. changes in MW may alter degradation in vivo and drug release kinetics. this can be accounted for and considered by making the initial MW higher or to conduct radiation sterilization while the product is refrigerated.

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37
Q

material considerations for radiation sterilization for biologics and human-based tissue

A

low dose radiation sterilization method, adding free radical scavengers or radioprotectors (antioxidant ascorbic acid)
ex. drug-material combinatorial products such as tissue-engineering scaffolds

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38
Q

chemical techniques

A

machines: bulk EO sterilization machine, tabletop sterilization machine
colorless gas, very toxic and flammable
requires special equipment with special venting requirements
low temperature sterilization method of choice for heat sensitive instruments: plastics, suture material, lenses, finely sharpened instruments
materials must be well aerated after sterilization
materials/instruments must be dry

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39
Q

material considerations for radiation sterilization: ethylene oxide

A

ethylene oxide functions on the basis of its strong alkylating property, causing disruption of cellular processes, including clotting of proteins, inactivation of enzymes, and disruption of DNA, resulting in preventing the replication of microorganisms.
preferred sterilized method for almost half the medical device manufacturing market. also effective for porous dressings and woven materials
liquids and powders are not compatible bc the gas has a hard time penetrating them unless spread out in a thin layer

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40
Q

other popular chemical agents

A

hydrogen peroxide and ozone
primarily leverage oxidation of the cell walls and proteins, resulting in the destruction of cell walls and even endospores with prolonged contact

41
Q

thermal sterilization

A

oldest class of sterilization technique. high temperature in both techniques (dry heat and steam) is the main principle of microbial inactivation. kill microorganisms by coagulation of proteins, including structural components of the cells, as well as by rupturing cell walls

42
Q

thermal techniques: dry heat

A

most convenient method for heat-stable materials such as nonaqueous materials, oil-based injectable pharmaceuticals, powders, glassware, and metallic surigcal instruments
uses hot air without added moisture to kill microbes
relies on temperature and exposure time (generally high temp (160-330C) for 3 h)
typically requires higher temp and longer exposure compared to steam sterilization

43
Q

thermal techniques: steam sterilization

A

sterilization by steam under pressure is relatively simple involving exposure of the product to steam at the desired temp and pressure.
carried out in a pressure vessel designed to withstand high temp and pressure
for uniform temp dist: important to remove the air from the sterilization chamber (accompanied by gravity displacement or by a vacuum system)
a vacuum system is generally preferred when compatible with the product/package system to ensure efficient air removal and optimum steam penetration

moist heat requires significantly reduced heat exposure time compared to dry heat and is nontoxic, rapid, penetrating, and energy efficient

44
Q

steam sterilization: autoclaving

A

autoclave: self locking machine that sterilizes with steam under pressure
sterilization achieved by the high temp that steam under pressure can reach
high pressure ensures saturation of wrapped surgical packs
ideal for metal instruments

typically a drying cycle for sterilizing items that need to be dry for immediate use or storage (surgical instruments, glass, metal tools)
drying cycle used for processing materials that would be compromised by residual moisutre

45
Q

steam sterilization advantages and disadvantages

A

advantages: simplicity, relatively short processing times, lack of toxic residues
a combination of high T with moisture has advantages over dry heat such as reduced sterilization time

disadvantage: relatively high T, humidity, high pressure may lead to softening, degradation, and hydrolysis of polymer-based materials. not effective at destroying endotoxin

46
Q

steam sterilization use

A

widely used in hospitals for sterilization of pharmaceuticals in glass ampules, plasticware, and metallic surgical instruments intended for reuse by applying saturates, pressurized steam
to be compatible with steam sterilization, a product must be stable with respect to temp and moisture, and the product/package must be readily penetrated by steam

47
Q

prion

A

misfolded protein that induces misfolding in normal variants of the same protein, leading to cellular death
heat-stable meaning it can survive sterilization procedures

48
Q

endotoxins

A

lipopolysaccharides found in the cell wall of gram-negative bacteria, which can induce inflammation and fever as an immune response in higher organisms
heat-stable meaning it can survive sterilization procedures

49
Q

blood coagulation (clotting)

A

process by which blood changes from a liquid to a gel, forming a blood clot. results in hemostasis, the cessation of blood loss from a damaged vessel, followed by repair
when artificial surfaces are placed in contact with blood, a complex set of interdependent reactions happen between the surface, platelets, and coagulation proteins

50
Q

which of the following is the primary goal of sterilization?
to kill all microorganisms, including spores
to reduce the microbial load to a safe level
to disinfect surfaces
to enhance the appearance of medical instruments

A

to kill all microorganisms, including spores

51
Q

which method is suitable for sterilization heat-sensitive medical equipment?
steam autoclave
dry heat sterilization
ethylene oxide gas
boiling

A

ethylene oxide gas

52
Q

in a standard steam autoclave, what is the typical temperature and pressure used to ensure effective sterilization?
100C and 10psi
121C and 15psi
160C and atmospheric pressure
180C and 30psi

A

121C and 15psi

53
Q

which statement best differentiates sterilization from disinfection?
sterilization uses steam, while disinfection uses dry heat
sterilization completely kills all microbes; disinfection reduces them
disinfection kills all bacterial spores; sterilization only reduces them
sterilization and disinfection are identical processes in infection control

A

sterilization completely kills all microbes; disinfection reduces them

54
Q

why is a drying cycle important in an autoclave process when sterilizing reusable biomedical equipment?
to enhance the effectiveness of dry heat sterilization
to remove moisture that could affect sterility and equipment function
to protect moisture-sensitive materials from damage
to maintain a high temperature for prolonged sterilization

A

to remove moisture that could affect sterility and equipment function

55
Q

true or false: dry heat sterilization better eliminates all microbes and spores on biomedical instruments, compared to steam

A

false

56
Q

true or false: ethylene oxide gas is often used to sterilize biomedical equipment that cannot tolerate high temperatures or moisture

A

true

57
Q

hemostatsis

A

process of blood clotting and then the subsequent dissolution of the clot, following repair of the injured tissue

crucial in regenerative medicine and tissue engineering bc it provides the initial step in wound healing. the hemostatic mechanism is designed to stop bleeding from injured blood vessels

58
Q

four stages of wound healing (7a slide 5)

A

hemostasis
inflammation
proliferation
remodeling

59
Q

cellular composition of blood: red cells

A

passive participants in the process of hemostasis
comprise 40-50% of the total blood volume

60
Q

cellular composition of blood: leukocytes (white cell)

A

perform many functions in inflammation, infection, wound healing, and the blood response to foreign materials, such as monocytes, neutrophils etc

61
Q

cellular composition of blood: platelets

A

nonnucleated, disk-shaped cells having a diameter of 2-3 um
platelets are produced in the bone marrow, circulate in the blood vessels, and occupy ~0.3% of the total blood volume

62
Q

platelet functions

A

initially arrest bleeding through the formation of platelet plugs
stabilize the initial platelet plugs by catalyzing coagulation reactions leading to the formation of fibrin

63
Q

platelet structure: platelet receptors

A

membrane-bound receptors that mediate the contact reactions of adhesion (platelet-surface interactions) and aggregation (platelet-platelet interactions)

64
Q

cytoplasmic storage granules

A

α-Granules, which are numerous and contain the platelet-specific proteins platelet factor 4 (PF-4) and β- thromboglobulin (β-TG), and proteins found in plasma (including fibrinogen, albumin, fibronectin, coagulation factors V and VIII).
dense granules contain adenosine diphosphate (ADP), calcium ions (Ca2+), and serotonin.
Lysosomal granules contain enzymes (acid hydrolases)

65
Q

Hemostasis events that occur in a set order following the loss of vascular integraity

A
  1. vasconstriction (vascular constriction). limits the flow of blood to the area of injury
  2. platelet activation. platelets activated by thrombin and aggregate at injury site, forming a temporary, loose platelet plug. the protein fibrinogen is primarily responsible for stimulating platelet clumping. platelets clump by binding to collagen that becomes exposed following rupture of the endothelial lining of vessels. Upon activation, platelets release ADP (Adenosine Diphosphate) and TXA2 (Thromboxane A2) (which activate additional platelets), serotonin, phospholipids, lipoproteins, and other proteins important for the coagulation cascade. In addition to induced secretion, activated platelets change their shape to accommodate the formation of the plug.
  3. blood clot. to ensure the stability of the initially loose platelet plug, a fibrin mesh (clot) forms and entraps the plug
  4. fibrinolysis & clot degradation. clot must be dissolved in order for normal blood flow to resume following tissue repair. dissolution of the clot occurs through the action of plasmin.
66
Q

platelet activation definition

A

causes platelets to become sticky and change to irregular spheres with spiny pseudopods

67
Q

platelet activation

A

platelets bind to matrix and spread to cover the damaged surface; aggregation to form temporary plug
- initiates the wound healing process through secretion of soluble small molecules from cytoplasmic granules called growth factors and cytokines, derived growth factor, fibronectin, von willebron factor, and transforming growth factor-beta
- these substances are sticky and bind to matrix, chemotactic (draw cells up the concentration gradient through migration) and/or mitogenic agents for leukocytes, endothelial cells and fibroblasts

68
Q

Hemostasis: Vasoconstriction & Plug Formation

A
  1. exposed collagen binds and activates platelets
  2. release of platelet factors
  3. attract more platelets
  4. aggregate into platelet plug
69
Q

fibrin clot formation - thrombogenesis

A

process by which a blood clot (thrombus) forms through the conversion of fibrinogen to fibrin by the action of thrombin
fibrin stabilizes the platelet plug and seals the site of injury to prevent further blood loss

70
Q

fibrin clot formation - thrombogenesis pathways

A

two main pathways: converse upon a final common pathway, which leads to the formation of thrombin and an insoluble fibrin gel when thrombin acts on fibrinogen
both pathways are complex and involve numerous proteolytic enzymes called clotting factors

71
Q

thrombogenesis intrinsic pathway

A

clot in response to an abnormal vessel wall superficial injury in the absence of tissue injury
longer and slower compared to extrinsic
can take between a few seconds or minutes to produce factor X
more Factor X produced than extrinsic

72
Q

thrombogenesis extrinsic pathway

A

clot formation in response to tissue injury, actual breakage of blood vessels
reacts almost instantaneously by producing factor X
main function is to augment the intrinsic pathway by slowing the flow of blood outside the vessel by producing little Factor X but quickly
completes the clot and allows for the blood vessel to be repaired

73
Q

coagulation & clot stabilization

A

clotting factors (proenzymes) interact in a sequential series of enzymatic activation reactions (coagulation cascade), leading to amplified production of the enzyme thrombin, which in turn cleaves fibrinogen to form a fibrin polymer that stabilizes the clot or thrombus

74
Q

platelet coagulant activity

A

when platelets aggregate, platelet coagulant activity is initiated including the expression of negatively charged membrane phospholipids, which accelerate two critical steps of the blood coagulation sequence: factor X activation and the conversion of prothrombin to thrombin

75
Q

thrombin

A

key enzyme that plays a central role in blood clotting. once formed from its proenzyme, prothrombin, thrombin not only converts fibrinogen into fibrin (to form a clot) but also activates other clotting factors

76
Q

activatable factors

A

clotting factors that exist in an inactive form (proenzymes or precursors) and need to be activated to participate in the clotting process. thrombin triggers their activation, allowing them to contribute to the formation and stabilization of a blood clot
factors like factor V,VIII,XIII are examples of clotting factors that can be activated by thrombin

77
Q

Factors V,VIII,XIII activation

A

Va: cofactor in the conversion of prothrombin to thrombin
VIIIa: plays role as cofactor in the intrinsic pathway
XIIIa: helps cross-link fibrin strands to stabilize the clot

78
Q

dissolving the clot and anticoagulants

A

the fibrinolytic system removes unwanted fibrin deposits to improve blood flow following thrombus formation and to facilitate the healing process after injury and inflammation

79
Q

plasmin

A

plasmin: most well-studied fibrinolytic enzyme, circulates in an inactive form as the protein plasminogen

80
Q

plasminogen

A

adheres to fibrin clot being incorporated into the mesh during polymerization
activated to plasmin by the actions of plasminogen activators, which may be present in blood or released from tissues, or which may be administered therapeutically

81
Q

Important plasminogen activators

A

tissue plasminogen activator (tPA) and urokinase

82
Q

following activation of plasminogen

A

plasmin digests the fibrin clot, releasing soluble fibrin-fibrinogen digestion products (FDP) into circulating blood, which may be assayed as markers of in vivo fibrinolysis

83
Q

fibrinolysis is inhibited by

A

plasminogen activator inhibitors (PAIs) and by a thrombin-activated fibrinolysis inhibitor (TAFI) which promotes the stabilization of fibrin and fibrin clots

84
Q

fibrinolytic sequence

A

Plasminogen activators, such as tissue plasminogen activator (tPA) or urokinase, activate plasminogen to form plasmin. Plasmin enzymatically cleaves insoluble fibrin polymers into soluble degradation products (FDPs), thereby effecting the removal of unnecessary fibrin clot.

85
Q

blood material interactions

A

There are two primary mechanisms through which thrombus formation is initiated from contact between blood and artificial materials:
1. interaction between platelets with the material surface, which is mediated by adsorbed proteins, leading to platelet activation
2. contact-activation of the intrinsic clotting cascade directly by the material surface

86
Q

thrombus formation induced by platelet adhesion on a synthetic material surface

A

(A) Adsorption of plasma proteins to material surface. (B) Conformational changes in adsorbed proteins followed by the adhesion of nonactivated platelets from the blood stream. (C) Platelet activation leading to the release of activating factors. (D) Platelet aggregation and the activation of the blood coagulation cascade leading to thrombus formation, along with the subsequent activation of complement and inflammatory processes.

87
Q

granulocyte

A

any blood cell containing specific granules (neutrophils, eosinophils, basophils)

88
Q

leukocyte

A

colorless blood cell capable of ameboid movement (lymphocytes, monocytes, granulocytes)

89
Q

macrophage

A

large phagocytic mononuclear cell

90
Q

ameboid

A

describes the adaptable, flowing movement of certain cells that allows them to move and interact with their surroundings, similar to the way an ameoba moves. they have the ability to change shape and move by extending and retracting pseudopodia (temporary projections of the cell membrane) to crawl or move

91
Q

what is the first step in the hemostasis process?

A

vasoconstriction

92
Q

which enzyme is responsible for converting fibrinogen to fibrin during clot formation?

A

thrombin

93
Q

what role does factor XIII play in the coagulation process

A

cross-links fibrins to stabilize the clot

94
Q

which of the following statements about platelets is true?
platelets are the main component of the clotting cascade
platelets adhere to the exposed collagen at the site of injury
platelets do not play a role in the release of clotting factors
platelets dissolve the fibrin clot after it is formed

A

platelets adhere to the exposed collagen at the site of injury

95
Q

what is the main function of plasmin in the hemostasis process?
to form the initial platelet plug
to activate thrombin
dissolve the fibrin clot during fibrinolysis
convert fibrinogen to fibrin

A

dissolve the fibrin clot during fibrinolysis

96
Q

true or false: thrombin is responsible for activating multiple clotting factors, including factor V and factor VIII

A

true

97
Q

true or false: fibrin degradation products (FDPs) are insoluble fragments formed during clot formation

A

false
soluble and formed during clot degradation

98
Q

true or false: the intrinsic and extrinsic pathways of the coagulation cascade converge at the activation of factor X

A

true