Test 2 Flashcards
guys i really dont get this class lmao
Biocompatibility
-Materials that elicit minimal adverse responses
Local response
An inflammatory defense that occurs at tissue level (seen in skin, mucosal tissue, epithelium)
Systemic response
An inflammatory defense that is spread throughout the body via blood or lympathic fluids. Less common with biomaterials, associated with lymphocytes.
Wear debris
- associated with ultra high molecular weight polyethylene implants
- Fragments are phagocytized by macrophages but can’t be digested, releasing lysosomal enzymes causing a loosening and failure of the implant
In vitro and In vivo
In vitro: testtube
In vivo: life
What the Body is doing to the Material:
1) Loosening
2) Phagocytosis
3) Corrosion/Wear/Tear
4) Fracture of implant/bone
5) Rejection of Implant
What the Material is doing to the Body:
1) Hemolysis/Thrombosis/Coagulation
2) Immunogenic
3) Inflammation
4) Toxicity
5) Tumorogenesis
6) Infection
7) Promotes encapsulation
Hemolysis/Thrombosis/Coagulation
Hemolysis: lysis and breaking open of red blood cells causing the release of hemoglobin into surrounding fluids
Thrombosis: a blood clot forms in circulatory system, impeding blood flow
Coagulation: clot formation
Inflammation
- An inflammatory cytokine response
- Beneficial: initiate tissue repair, rid body of harmful agents
- Deleterious: may have over active response to minor agent or self, autoimmune disease
- Acute vs Chronic
Bacterial Biofilms
Adherent bacteria for a glycocalyx coat, this coat protects bacteria from phagocytosis and antiobiotics = bad
Encapsulation
- The coating or engulfing of particles within a continuous matrix
- Many implants trigger fibrous encapsulations, which confines the foreign material and keeps it separated from surrounding normal tissue
Phases of Wound Healing w/ Biomaterials
1) Hemostasis: stop bleeding (platelets, fibrin)
2) Inflammation: initiates wound repair (neutrophils, macrophages, lymphocytes)
3) Proliferation/Granulation: pulls the wound close, beginning of scar formation (fibroblast, epithelial cells, endothelial cells)
4) Fibrosis: formation of fibrous connective tissue surrounding the foreign object (fibroblasts)
5) Foreign body reaction: phagocytosis occurs of foreign object (FBGC, fused macrophages)
6) Remodelling/Maturation” development of the final proper tissue (everything)
In vitro tests
Compared to in vivo tests, these tests are high-throughput, fast, inexpensive, not always relevant to in vivo results
Kilgman Maximization Test
-Determine allergic or sensitizing capacity to repated or prolonged exposure of agent
-Characterized by delayed reaction, independent of dose
-Evaluates a system response, often seen in skin
Steps: intradermal injection in guinea pigs, 7-9 weeks later, inject agent/material elsewhere, then look for reaction on skin
Irritation Tests
Evaluates a local tissue inflammation, usually seek in skin
Intradermal injection into albino rabbit
Genotoxicity Test
- Evaluates genotoxicity effects, DNA destruction, gene mutation, and chromosomal abnormalities
- Uses a mutation reverting bacteria that does not produce histidine (Salmonella)
- Uses media that lacks histidine
- A mutagen may cause bacteria to grow without external source of histidine
- The mutagenicity of agent = the number of bacterial colonies in media
Prediction of Cell Yields
- cell yield on the day of surgery is a critical factor
- seeding cell desnity or cell number is commonly critically defined in most cell therapy protocols to assure a certain therapeutic effect
Serum-Free conditions
- ex vivo expansion
- human serum not realistic
- FBS: -possible risk of contamination, immunological reactions against xenogeneic serum
Scaling-up Cell Microencapsulation
Transgenic Cell Encapsulation > Encapsulated Cell Culture > Transplantation
- Follow GMP procedures
- Scaling-up cell encapsulation: 3D air supply and multinozzle outlet
- Once therapy reached goal / when undeseriable deleterious effects occur, noninvasive
- monitoring and deactionation/elimination of the encapsulated cells are critical
Cell Storage - Cryoprotectants
- Minimize damage caused by ice crystal formation, cytotoxic to some extent
- Scaled-up cryopreservation for macroscopic tissue problems related to heat and mass transfer
- Large products need longer incubation time with cryoprotectants = lower survival rate
Cell Storage - Vitrification
Prevents ice formation throughout the entire sample during cooling and warming process
Cell Storage - Dry Storage
- long-term unfrozen storage at ambient temperature
- Low cost (not yet a reality)
- Many organisms can undergo anhydrobiosis to survie in a completely dehydrated state for an extended time and resume activity upon rehydration
Determination of Cell Fate
- Biological, Chemical, Mechanical, Physical Properties
- Stimuli affecting stem cell differentation
Biological Properties
- Growth factors
- Genetic material
Chemical Properties
- O2
- Free radicals
- Contact angle
Surface Hydrophobicity
- Assessed by measuring contact angle through water spread on droplet on a surface
- The lower the contact angle, the more hydrophilic the surface is
- Superhydrophilic (<5), Hydrophilic (<90), Hydrophobic(90-150), Superhydrophobic(150-160)
-Maximum adhesion of fibroblasts: 60 - 80 degrees, for osteoblasts: 0 - 106 degrees
Surface Charge
- Increased polarity, increased migration
- Negative charge, increase chondrocyte differentation
- Positive charge: improve neuronal attachment, osteoblasts & fibroblasts spreading
Cell Responses to Surface Chemistry
- Immediately after scaffold implantation, protein adsorption occurs, which mediates cell adhesion
- Integrins affect ECM deposition, cell proliferation, and differentation
- Focal adhesion: interaction between cells and biomaterial scaffold
Mechanical Properties
- Shear
- Stiffness
Static vs. Shear (Dynamic)
Static Culture: nonhomogenous growth, nonhomogenous differentation, low O2 and nutrient diffusion, difficulty of monitoring and control, low productivity
Dynamic Culture: better homogeneity, O2 and nutrient supply during exposition to shear stress
- Higher cellular growth
- Higher control and productivity
Tissue’s Young Modulus
- Given by resistance offered by the tissues to deformation effects, i.e. the tissue stiffness
- Use rheometer to measure cell adhesion force on substrate softness