MS4013: Biomaterials

L01 – Introduction to Biomaterials

Biomaterial VMB
Non-viable material used in a medical device intended to interact with biological systems
Non-viable: not from living organism
e.g. glasses (non-viable, medical device, but not interact with biosystems)
e.g. contact lens (non-viable, medical device, interact with biosystems)
Biomaterial Biological material
Origin Natural/synthetic Living organism
Application Biomedical/biotechnological Any, e.g. biomaterial
Example PMMA, Nitinol Fibrin, Collagen
Biomaterial: nano/micro/macro
Pharmaceutical: atomic/molecular

Biocompatibility PAH
Ability of a material to perform an appropriate hose response in a specific application
2 factors:
 Effect of the implant material on the body
 Effect of the body on the implant material

Application of Biomaterials (length of usage) TDP

 Temporary – plasters, contact lenses
Contact lense can react with protein in eye  blur vision  need to remove and wash
 Degradable – sutures
Need to be strong enough in the beginning and degrade later
 Permanent – pace markers, joint replacements
Need to be strong permanently

Application of Biomaterials (usage) ITD

 Implants – Orthopedic (largest and fastest growing), pacing device (cardiovascular device),
neurostimulators, drug implants
 Tissue engineering – multidisciplinary (biology, material, physics)
 Drug delivery – e.g. lessen side effect of tumor targeting drugs

Biomaterials MCPC
 Metals TSAM
 Ti, Ti alloys, stainless steel – joint replacements/fixation plates (strength)
 Stainless steel – pacemaker wires (conduct electricity)
 Amalgram – dental fillings (multiple metal, include mercury for malleability)
 TiNi shape memory alloy, stainless steel – dental wires
 Ceramic ACPB
 Aluminum oxide (Al2O3) – joint replacements (less wear)
 Calcium phosphate (Ca3(PO4)2) – bone graft substitutes, bioactive coatings (fixed implant to bone)
 Pyrolytic carbon – surface coatings, reinforcing fibers
Antithrombotic: reduce formation of blood cloth (thrombi), used as coating in heart valves
 Bioactive glasses – bioactive coatings, implants, attachment
 Polymer PSPN
 Polyethylene – joint replacements (cushioning)
 Silicone – breat implants, small joint replacements
 Polylactic acid/polyglycolic acid – degradable sutures, degradable implants
  Nylon – non degradable sutures
 Composite PFS
 Particulate-reinforced – dental fillings, middle ear implants
 Fiber-reinforced – fracture fixation, ligament replacement
 Self-reinforced – fracture fixation

Joint replacement: hip implant CLHS

Cup (metal) – strength, porous surface
Liner (ceramic) – wear resistant
Head (ceramic) – wear resistant
Stem (metal) – strength, porous surface
Cushion (polymer)

Effects of implant on body IAT

 Bioinert – induce capsule response, ignored, not harmful, isolated
 Bioactive – encourages an advantageous response from the body (e.g. bonding)
 Toxic – kills cells/tissue in contact with/away from implant

Effects of body on implant EDCP

 Environmental – body fluid 0.9% saline, cells, and protein
 Degradation – intentional/unintentional
 Corrosion – mainly metals
 Protein absorption – mainly polymer (contact lens)

Implant failure ILFWA

 Infection – from contamination (bacteria/virus), need to remove
implant to allow antibiotics to work
 Loosening – leads to pain and loss of function
 (Mechanical) Fracture and wear – increases with time

 Adverse responses
 Contraction – breast implant not pressure resistant, can cause scar tissue and harden
 Reaction to particles/wear – in joint implants, immunity can remove coating and release particle
and more wear

Factors Influencing PSM

 Condition of patient
 Competence of surgeon
 Compatibility of material

History
 Copper sterilizer for sutures  allow surgery without implanting bacteria
 Post WWII chaos in biomaterials world
 Materials that had been rationed were now available
 Surgeons did not collaborate with scientist/engineer
 Dentist/doctors invent devices “on the fly” when patient’s lives or functionally were at stake
 Minimal government/regulatory oversight  unmonitored work
 Thalidomide tragedy
 After 1950s
 Formal regulatory process
SG: Health Science Authority (HSA)
US: Food and Drug Administration (FDA)
China: China Food and Drug Administration (CFDA)
Canada: Canadian Medical Device Regulations (CMDR)
Europe: CE
India: Drug Controller General of India (DCGI)
 Fluorescence microscopy can understand biocompatibility on a cellular and molecular level

Development of medical devices NDBETACL

 Identify a need
 Design of medical device
 Biomaterials research and testing of
 Engineering to develop a medical device
 Preclinical and clinical testing
 Regulatory approval
 Commercialization and clinical application
 Long-term follow up and explant analysis

Important aspect when designing biomaterials

Design consideration FEH
 Function of device
 Physiological environment
 Healing and regeneration process
Materials selection BPDPCAR
 Biocompatibility
 Physical and chemical properties
 Durability and degradation rate
 Production, sterilization, packaging
 Cost and availability
 Prior approved usage
 Risk analysis

Changing role of biomaterials ASB

Passive  Interactive  Integrative  Instructive:
 More aggressive role
 Recruits specific material-tissue interactions
 Forms bond between tissue and material
Replace (e.g. THR)  Repair (e.g. metal fixation plates)  Regenerate (e.g. scaffold with cells)

Logic of tissue-inspired engineering LEC

 Role of material scientist: leverage on biological/anatomical knowledge  design environments 
enable cells to perform their engineering role (cells as the “tissue engineers”)

Tissue Engineering IPELBRMIT

Interdisciplinary field that applies the principles of engineering and life sciences towards to development of
biological substitute that restore, maintain, or improve tissue function

Why tissue engineering ORU

 Shortage of whole organs – TE: create new organ/regenerate the damaged one
 f4 organ high demand: kidney, liver, heart, cornea
 Limited organ regeneration potential
High: skin, intestine, liver SIL
Moderate: bone, muscle BM
Low: heart, brain, cartilage (requires vascularization (excessive formation of blood)) HBC
 Unmet medical needs

Current therapies, types of graft TAAXM

 Transplantation
 Autografting (same patient) - TE
 Allografting (other living or diseased donor) - TE
 Xenografting – tissue from animal source (e.g. porcine and bovine) - TE
 Man-made, biomimetic materials and devices – permanent solutions

Market potential
 Global regenerative medicine market grow 20.7% during 2016-2020 and reach $49.41bn
 Biggest: therapies for degenerative and traumatic orthopedic and spine applications
 TE product success on market: skin and cartilage

Tissue engineering endpoints CAFC

 Morphology/histology/biochemical – match the composition and architecture of the tissue
 Functional – achieve certain functions, display certain properties (e.g. mechanical)
 Clinical – pain relief, extension of life

Biological material
Advantages CDMS Limitation ITCV
Biocompatibility Immunogenicity
Biodegradability Temporary
Biomimetic Complex
Susatainable Variability

L02 – Structure and Organization of Cells and Tissues

Levels of organization: Cells  Tissues  Organs  Systems  Organisms CTOSO

Organism IRRGH
An individual living thing that is capable of response to stimuli, reproduces, grows, and maintains
homeostatis.
Taxonomy: Hierarchical system to classify and identify organisms

Classification of organism EP
 Eukaryote (animal, plant, fungi, protist) APFP
 Multicellular organisms
 Have membrane-bound cell nucleus and organelles
 Prokaryote (bacteria, archaea) BA
 Unicellular organisms
 No membrane-bound cell nucleus and organelles
Virus??

System FOSIH TPGB

Several function-related organs which together perform a continuous physiological function
 Each located in specific location with specific function
 Many internal organ systems enclosed within coelom (cavity within body), e.g. digestive system
 Function of organ system is to contribute to maintain homeostasis (stable internal environment e.g.
temp, pH, glucose, blood pressure)

Various Systems MRHERCIND

 Muscular and skeletal – support and movement
 Respiratory – gas exchange
 Hormonal – regulation of internal environment and development
 Excretory – get rid of metabolic wastes
 Reproductive – producing offspring
 Circulatory – transport of necessary molecules to cells
 Immune – defense against invading pathogens
 Nervous and sensory – regulation and control, response to stimuli, information processing
 Digestive – convert food to usable nutrients

Organ TF EDS
Different tissues joined and working together, made of one/more types of tissues (79 organs in human)
Functional grouping together of multiple tissues
Largest organ: Skin (3 layers)
1. Epidermis (protection) MK
 Melanocytes (melanin): gives pigment to skin
 Keratinocytes (keratin): gives hardness to skin cells
 Langerhans cells and Merkel cells
2. Dermis (connective tissue, glands) FESA
 Fibroblasts, endothelial cells
 Hair follicles, sweat glands, sebaceous glands, apocrine glands
 Matrix components
3. Subcutaneous layer (connective tissue, fats) AM
 Adipocytes: fat cells
 Macrophages

Tissues AIOF
 Assembly of cells, not necessarily identical, same origin, together carry out specific function
 Histology: study of tissues

Classification based on function organ

 Protective tissues – e.g. skin
 Mechano-sensitive tissues – e.g. bone, ligament, tendon
 Electro-active tissues – e.g. brain, skeletal muscle, heart
 Shear stress-sensitive tissues – e.g. blood vessels

Types of tissues ECMN

1. Epithelial CC SCC
 Covers body surfaces and line body cavities
 Outer epidermis (skin) protects from injury and drying out
  Inner epidermis (internal surface) protects, secretes mucus
 Made of closely packed cells (interior epithelium covered with a mucus layer)
 3 types
 Squamous epithelium – flat shaped
 Function: protection, diffusion, osmosis, filtration PDOF
 Where: alveoli, capillary walls, blood vessels, oral cavity
 Cuboid epithelium – cube shaped
 Function: protection, secretion, absorption PSA
 Where: line kidney tubules (filtration), surface of ovaries, sweat glands
 Columnar epithelium – column shaped
 Often have microvilli or cilia
 Function: absorption, movement of something along surface AM
 Where: lining of intestine, oviduct, uterus
 Can exist as single layer or stratified (layer stacked on each other)
Simple squamous Stratified squamous
Exchange nutrient, waste, gas Protect abrasion, drying out, infection
Lines blood vessels, capillary walls, alveoli Outer layer of skin, mouth, vagina
Simple cuboid Stratified cuboid
Secretes & reabsorbs water & small molecules Secretes water & ions
Line kidney tubules Line ducts of sweat glands
Simple columnar Stratified columnar
Absorbs nutrient, produces mucus Secrete mucus
Lines most digestive organ Lines epididymis, mammary glands, larynx

2. Connective BSPF LFCBB

 Binds and supports body parts structure together
 Fill up spaces
 Provide protection
 Store fat
 5 types
 Loose – join tissues, hold organs in place, e.g. lipid (fat storage)
 Fibrous – bundles of collagen fibers (very strong), e.g. tendons (muscle-bone) and ligaments
(bone-joints)
 Cartilage – flexible matrix rich in protein and fibers, e.g. nose, ears, vertebrae, ends of bones
 Bones – rigid connective tissue, extracellular matrix mineralized by calcium phosphate
 Blood and blood forming tissues – connects body system together (oxygen, nutrient,
hormones), same origin as other connective tissue (mesodermal), 55% plasma (liquid matrix),
45% erythrocytes
3. Muscular AMR SSC
 Composed of muscle fibers containing actin and myosin proteins
 Interaction responsible for movement
 3 types
 Smooth: non striated, involuntary control, contract slowly, contract longer time
internal organs, intestine, stomach, blood vessels
 Skeletal: striated, voluntary control, contract quickly and strong, can fatigue
attached to bones used for movement
 Cardiac: striated, involuntary, contract quickly, beats whole life
heart

4. Nervous NBS ER
 Specialized tissue that forms nerves, brain, spinal cord
 Conducts electrical and chemical signals along neuron
  Responds to stimuli and transmits impulses from one body part to another
Nerve fibers:
Dendrite: conduct signal to cell body
Axon: send signal away from cell body
Bundle of nerve fibers (dendrite + axon)  nerves

Nerves conduct signal to and from, brain, spinal cord, and sense organ
to register sensation and trigger muscle movement

Artificial tissues and organs

 Artificial heart – made of synthetic material except the valve in the middle from biological material
 Artificial hand with feelings – involves electrical engineering to restore sensation
 Artificial blood vessels – must have elasticity and tensile strength to withstand blood pressure
 Artificial trachea – must be air-tight and leak proof, tissue engineering and scaffold
 Artificial kidney – specific function of filtration
 Artificial bladder – cell culture media containing patient’s cell and biodegradable bladder scaffold

Challenge of organ printing BCV

 Biomaterials: availability and compatibility
 Cells: hard to grow outside, especially liver and pancreas
 Vascularity: supply of blood to allow organ survive and regenerate

3D printing
 3D file sliced to 2D slices that are printed layer by layer
 96% of hearing aids are 3D printed
 Reduce surgery time from 97 to 23 hours
 Bone printing, bronchus printing
 Still difficult to 3D print biofunctional organ

Cell BSFEP
 Basic structural and functional unit of life
 2 types of cells and organism: eukaryotic (multicellular) & prokaryotic (unicellular)

 >220 types of cells in the body, 1 cell type alone does not allow formation of tissue/organ

Animal Cell MNC

 Membrane (surface) WP
 Cell wall PTEPS
 Commonly found in plant
 Controls turgidity
 Extracellular structure surrounding plasma membrane
 Primary wall cell is extremely elastic
 Secondary wall cell forms around primary wall cell after growth is complete
 Plasma membrane OTPP
 Outer membrane of cell
 Controls cellular traffic
 Contains proteins, span through the membrane and allow passage of materials
  Proteins are surrounded by a phospholipid bi-layer
 Nucleus CNN
 Chromosomes CDTS
 Usually in the form of chromatin
 Composed of DNA, contains genetic information
 Thicken for cellular division
 Set number per species (i.e. 23 pairs for human)
 Nuclear membrane NLO
 Surrounds nucleus
 Composed of 2 layers
 Numerous openings for nuclear traffic
 Nucleolus SVR
 Spherical shape
 Visible when cell is not dividing
 Contains RNA for protein manufacture
 Cytoplasm GERMCCLV
 Golgi apparatus PMN
 Protein ‘packaging plant’
 A membrane structure found near nucleus
 Composed of numerous layers forming a sac
 Endoplasmic reticulum TCSSR
 Tubular network fused to nuclear membrane
 Goes through cytoplasm onto cell membrane
 Stores, separates, and serves as cell’s transport system
 Smooth type: lacks ribosomes
 Rough type: ribosomes embedded in surface
 Ribosomes TFSM
 Each cell contains thousands (25% of cell’s mass)
 Miniature ‘protein factories’
 Stationary type: embedded in rough endoplasmic reticulum
 Mobile type: injects protein directly into cytoplasm
 Mitochondria LDCER
 Second largest organelle with unique genetic structure
 Double-layered outer membrane with inner folds (cristae)
 Controls level of water and other materials in cell
 Energy-producing chemical reactions at cristae
 Recycles and decomposes proteins, fats, and carbohydrates, and forms urea
 Centrioles PTC
 Paired cylindrical organelles near nucleus, lie at right angles to each other
 Composed of 9 tubes with 3 tubules each
 Involved in cellular division
 Cytoskeleton MSA
 Composed of microtubules
 Supports cell and provides shape
 Aids movement of materials in and out of cells
 Lysosomes DTSP
 Digestive ‘plant’ for proteins, fat, and carbohydrates
 Transports undigested material to cell membrane for removal
 Vary in shape depending on process being carried out
 Cell breaks down if lysosome explodes (if there is pH difference)
 Vacuoles MWC
 Membrane-bound sacs for storage, digestion, and waste removal
  Contains water solution
 Contractile vacuoles for water removal (in unicellular organisms)

L03 – Host Response to Materials

Host response/biocompatibility
 Material-tissue interaction IRW
 Implant-tissue interface
 Evolution: protein adsorption, cell attachment
 Role of surface properties
 Response to wear particles and metal ions
 Wound healing
 Foreign body reaction, fibrous capsule formation
 Determination of biocompatibility
 IO 10993 biological testing matrix
 In vitro and in vivo testing

Host Defense CPI

 Body under constant attack by microorganism in the environment
 Pathogen: an infectious agent that causes disease
Infectious disease occurs when a microorganism succeeds in evading or overwhelming host
defense to establish a local site of infection and replication
 Defense EI
 1st line of defense: epithelial tissue (physical defense)
 2nd line of defense: immune system (immunal defense)
Bacteria releases protein  alert macrophages  attach to bacteria  protein: surface
reaction  bacteria locked and digested inside phagosome PMASD

Material-Tissue Interactions
1. Implant-tissue interface ??
Evolution of the interface (dental implant-bone interface)

~weeks: no more inflammation

~10 years: full recovery

Interface (different length scales) IBCP

Proteins, cells, organized tissue strongly influenced by implant surface structure that same length of scale
 Macroscale: (implant-bone) implant shape need to fit
 Microscale: (implant-cell) affect cell interaction
 Nanoscale: (implant-protein) topography affect cell behavior and bone formation at the implant
surface

Evolution of interface (time progression of events) IWPCF

 Implant
 Water adsorb to surface
 Proteins adsorb to surface:
adhesion of serum and membrane
protein (integrin)
 Cell attach
 Fibrous tissue develops

Influence of surface chemical properties PWC TCWC

 Protein adhesion
 Water adsorption
 Cell attachment (cell interact with the protein coated surface, not the
material surface itself)

Biomaterial surface properties

 Surface topography/roughness
 Surface charge – hydrophilic: encourage cell growth better as there is
more contact area with water on surface)
 Surface wettability (polarity) – no water, less protein, less cell attach
 Surface chemical composition

Properties measurement techniques ??

Composition ESCA, SIMS, NEXAFS, Auger
Structure ESCA, SIMS, NEXAFS, FTIR, SFG
Orientation NEXAFS, FTIR, SFG
Spatial distribution SIMS, AFM
Topography AFM
Thickness ESCA, AFM, Ellipsometry, SPR
Energetics Contact angle

Cell attachment trigger different cell fate processes SE

 Successful implant, the protein-coated biomaterial must support these
functions (biological outcomes) VPC PMDA
 Viability
 Protein synthesis
 Communication
 Proliferation (certain cells)
 Migration (certain cells)
 Differentiation/activation (certain cells)
  Apoptosis (certain cells)
 Depends on protein shape/what the protein will exposed
 (A) not very exposed, cells harder to attach (can attach but not develop)
 (C) fully spread, higher survivability and can develop

2. Wound healing
Wound healing without implant ICELRR
a. Injury
b. Coagulation
c. Early inflammation
d. Late inflammation
e. Repair
f. Remodeling

Wound Healing Process CIRR

a) Blood coagulation BA
 Formation of blood clot (fibrin + platelets) stops blood loss from the body
 Activated platelets and blood clot release signaling molecules, attract inflammatory cells to the wound

b) Inflammatory phase RDS

 Remove cellular and tissue debris from the wound
 Destroy any foreign objects (e.g. microorganism)
 Secrete signaling molecules, attract new cells to the wound to produce new tissue

Chemotaxis (how cells communicate) MLC BPBL

 Cells migrate up a concentration gradient of molecules to the source of the molecule
 Help leukocytes to find site of injury or infection
 Chemotactic molecules for leukocytes: (“called” to the site of tissue injury and blood clot formation)
 Bacterial peptides
 Molecules from activated platelets
 Molecules from blood coagulation
 Molecules from other activated leukocytes

Elements of blood RPW MLG BEN

 Red blood cells
 Platelets
 White blood cells (leukocytes)
 Inflammatory cells, circulate in blood stream and migrate to tissue to kill/consume/destroy foreign
objects
 Include:
 Monocytes
 Lymphocytes
 Granulocytes (eosinophils, basophils, neutrophils): have internal granules, also called
polymorphonuclear (PMN) due to lobed shaped nuclei

Neutrophils FAP
 One of the first cells to arrive, migrate from bloodstream to tissue
 Involved in acute(initial/early) phase of inflammatory response
 Capable of phagocytosis

Monocytes/macrophages ICP
 Monocytes are immature cells until leave bloodstream. Migrate into tissue, turn into macrophages
(swell and become granular)
 Macrophages involved in the chronic (later/long term) phase of inflammatory response
 Capable of phagocytosis

Phagocytosis CIPLERD
1. Chemotaxis and adherence of microbe to phagocyte
2. Ingestion of microbe by phagocyte
3. Formation of phagosome
4. Fusion phagosome + lysosome  phagolysosome
5. Enzyme digest of ingested microbe
6. Residual body containing indigestible material
7. Discharge of waste material

c) Repair phase GFE

 Formation of granulation tissue to fill the wound space
 Fibroblast cell recruitment, proliferation & production of extracellular matrix (ECM)
 Endothelial cell recruitment, formation of new capillaries and blood vessels (angiogenesis)

d) Remodeling phase SF
 Granulation tissue remodeled to scar tissue, and eventually to proper tissue
 Foreign body reaction and fibrous capsule formation around implants

Wound healing with implant FC

 The only difference: formation of fibrous capsule around the implant during the chronic phase

Foreign body reaction to implant FF

Presence of macrophages and foreign body giant cells (FBGC) at implant surface when implant in the body

FBGC FMF
Result of frustrated phagocytosis (cannot remove) 
macrophages fuse together to form large cells to
ingest foreign objects  FBGC formed

Fibrous capsule response FFF

 Frustrated phagocytosis
 FBGC formation
 Fibrous capsule formation CFTIS
 Layers of collagen and fibroblast
 Continuous recruitment of fibroblasts cells  building up of tissue and ECM
 Thick tissue prevent blood vessel development, cannot be remodeled
 Inflammatory phase that never ends
 Capsule surround implant to isolate interaction

3. Response to wear particles and metal ions FOM

a) Frustrated phagocytosis LIP
 Caused by large/many/high aspect ratio/high surface biomaterials/fibers/particulates
 Cells cannot completely ingest foreign object  frustrated phagocytosis
 Contents of phagolysosomes spill out into ECM and implant surface  kills phagocyte  attract new
macrophages  try to digest  process repeats

b) Osteolysis (specific in bone) RDC

 Bone resorption due to high number of wear particles
 Dose dependent response
 Result: continued chronic inflammatory response (macrophage try to remove particle but cannot)

c) Metal ion hypersensitivity HIP RDT

 Released ions from metallic implants act as haptens
(complex with native protein, activate immune response via B and T cells)
 Release ion  phagocyte cannot differentiate  no antibodies produced
 Ion stick to other protein  immune cell detect  antibody produce (attack
protein)  auto immune (hypersensitivity: undesirable/excessive immune
response)
 Hypersensitivity reaction (allergy, type IV)
 Delayed-type hypersensitivity
 24-72 hours after second contact with antigen
 Contact dermatitis: Cr, Co, Ni ions
 Deep tissues: metallic, silicone, acrylic implants (small oligomers from polymers act as haptens)

??

Biocompatibility ATM
 Biocompatibility: ability of material to perform an appropriate host response in a specific application
 Biocompatibility controlled by tissue response
 Tissue response controlled by material surface (unless controlled release of bioactive substances from
the biomaterial, e.g. bioactive materials)

Biocompatible materials for material-tissue interaction BMT IITT

 Biomaterial, medical device, tissue engineering construct that can be brought into direct contact with
living tissue and:
 Cause no prolonged inflammatory response/immune response that could compromise function
 Cause no local/systemic toxic reaction
 Have no tumorigenic potential

Testing method IPO IHD RNTC

 In vitro
 Cell culture polystyrene (surface modified polymer)  readily attach and grow most cells in culture
Untreated polystyrene will neither attach/grow mammalian cells
 One cell with one material
 In vivo
 Both materials heal almost indistinguishably with a thin foreign body capsule
 More dynamic, not fixed system
 Result of the in vitro test do not provide all information relevant to the implant situation.
Need in vitro (simple) and in vivo (dynamic) system
 Clinical implication
 Nontoxic in in vitro does not mean nontoxic in vivo
 Toxic in in vitro does not mean cannot be used (e.g. unique application, best clinical efficacy)
 Clinical acceptability depends on several factors, toxicity is only one of it

ISO 10993 Series I

 International harmonized standard to determine biocompatibility of device and material
 Testing procedure for CCD BII
 Chemical test and material characterization
 Characterization of material structure and composition (bulk and surface)
 Determination of degradation product and leachable substance and
 Biological test
 In vitro
 In vivo
 Testing on FMD
 Sterile final product or sample from final product
 Materials processed and sterilized in same manner as final product
 Degradation products and leachable extracts
 Category of medical device based on ND
 Nature of body contact
 Contact duration
 Determines extent of biological testing (FDA vs ISO tests)
 Selection of tests based on intended use and risk assessment
 In vitro first, then in vivo
 Negative and positive control should be included

Types of tests IBICI

 In vitro bioactivity
 Chemical response of material to body fluids
 In vitro cytocompatibility
 Cell response to elutant from materials soaked in different media
 Cell response to contact with material
 In vivo
 Implantation – provides both local tissue response and whole body response

In vitro assessment for cytocompatibility ECCP

 Evaluation of biomaterials by methods that use isolated, adherent cells in culture to measure
cytotoxicity and cytocompatibility
 Cytotoxicity – what is toxic for cell at cellular level (death, alteration in cellular, enzymatic inhibition)
Cytocompatibility – not specifically kill cell, but cause problem
 Cytotoxicity different from physical factors that affect cellular adhesion

In vitro assay methods EDAM

 3 primary cell culture for biocompatibility
 Elution (extract dilution) – more for cytotoxicity
 Direct contact – more for cytocompatibility
 Agar diffusion – more for cytocompatibility
 Morphological assays: outcome is measured by observations of changes in morphology of the cells

Elution testing SECDM

 Soak sterile sample in sterile tissue culture medium for 24/48hrs  get elutant
 Culture cells in 100%, 50%, 10%, 1%, 0.1% elutant and look at cell survival
 How much does the elutant need to be diluted to not to kill the cells
 Cytotoxic if <70% cells alive or kill >30%
 Material soak in medium  only test the medium, not the material itself

Contact testing PCB

 Can be put above/below
 More for cytocompatibility, to test when in contact with material
 Brownish outer ring  dead cells  not cytocompatible

In vitro testing
Advantages STAHREC Disadvantages LMI
Simple (cells) No loading (simple) e.g. no hip implant
Can choose cell type No motion (simple) e.g. no wear
Use cell line or primary cell (accessible) Individual cells, not system
Can use human cells
Moderately repeatable
Ethics and cost (only using cells)

Goal of In vivo testing EEP

 Exploratory – understanding of fundamental biological mechanisms
 Explanatory – understand complex biological problems (how material kill cells)
 Predictive – discover and quantify the impact of investigation

Factor to consider – Animal model RPEAC

Regulated by IACUC (Institutional Animal Care and Use Committee)
 Research factors
 Physical and environmental factors
 Animal related factors
 Animal care factors
Animal models and biomaterial applications ??
 Cardiovascular: sheep (heart valves), calves (artificial heart)
 Neurology: rat, non-human primates (peripheral system)
 Ophthalmology: rabbit, monkey (eyesight)
 Orthopedics: sheep, goat (close to human)
 Respiratory system and urogenital track: dogs, pigs (same size)
 Wound healing: young pigs (identical 11 hair follicles/cm2)
 Dental application: dogs, monkeys, pigs (identical tissue response, all omnivorous)
 Otology: cats, chinchilla (same bones and inner ear structure)
 Sometimes work in human but not in animal

Important material considerations FT

 Fulfill their intended function (some in seconds some lifetime)
 Lifetime animal < human  cannot always test for the duration of the usage, need to predict what
happen

Components relevant to in vivo assessment of tissue compatibility MADLOF

 Material of manufacture
 Intended additives, process contaminants, and residues
 Degradation products
 Leachable substance
 Other components and their interactions in the final product
 The properties and characteristics of the final product

In vivo tests for tissue compatibility ??

 Sensitization
 Irritation
 Intracutaneous reactivity
 System toxicity (acute toxicity)
 Subchronic toxicity (subacute toxicity)
 Genotoxicity
 Implantation
 Hemocompatibility
 Chronic toxicity
 Carcinogenicity
 Reproductive and developmental toxicity
 Biodegradation
 Immune responses

Animal Model RCRP

 Animal-related factors LASH
 Life-span
 Age
 Sex
 Health condition
 Animal care factors CHEP
 Cost
 Housing
 Euthanasia (sacrificial), need to reduce animal testing
 Pre/post-operative care (if surgery)
 Research factors FTDS
  Intended function
 Time required
 Device category
 System involved
 Physical and environmental factors CCLR
 Composition
 Characterization
 Leachable/degradation products
 Responses

In vivo testing IRHEM

 Implant in an animal, after defined times animal is killed
 Remove section (remove implant and/or tissue around it to see what happen)
 Histology, electron microscopy, mechanical testing, growth labels

Position of implantation SII

 Subcutaneous (below skin) or Intraperitoneal (inside abdominal cavity)
 Below skin  easy to implant/get
 Range of materials can be implanted
 In situ (in that position itself)
 Can be more difficult to implant
 Is in the appropriate tissue and dynamic environment (e.g. cardiovascular)

In vivo testing
Advantages WD Disadvantages ECRAPL
Whole system Ethics and cost
Dynamic system Relevance of species used
Animal to animal variation
Not a person
National laws
TVSREC
In vitro vs In vivo: target selectivity, experimental variability, system representation, regulatory control,
ethical, cost. All higher in vivo except target selectivity

L04 – Application 1: Tissue Engineering

Tissue engineering RFI DFL
Domain of regenerative medicine, goal is the development of functional tissues and organs (in vitro) for
implantation (in vivo) or direct remodeling and regeneration of tissue (in vivo) to repair, replace, preserve,
or enhance tissue or organ function lost due to disease, injury, or aging
Regenerative medicine RRF
Replaces or regenerates human cells, tissues, or organs, to restore or establish normal function
IELBFO
Interdisciplinary field that applies principles of engineering and life sciences toward the development of
biological substitutes that restore, maintain, or improve tissue function or a whole organ.

4 Factor of TE: CEBP

1. Cells to do the job – from target organ/stem cells/lab grown
2. Environment to support cells – from donor tissue/actual or synthetic polymer
3. Biomolecules to make cell healthy and productive – add directly/from cells at scaffold
4. Physical and mechanical forces to influence development of cells

TE method BICSRE
1. Biopsy and cell isolation
2. Cell cultivation
3. Cell seeding onto 3D biomaterial scaffolds and addition of biomolecules
4. Simulation of body conditions using a bioreactor
5. Engineered tissue for transplantation treatment

TE processes strategies TCLPI

1. Tissue induction
Scaffold directly implanted (in vivo)  cell
from body migrate inside and colonize 
grow, develop, fill gap, blood vessels

Not for any tissue, good for bones

2. Cell transplantation
In vitro scaffold, already have pre-cultured
cells  cells attach and develop before in body

Sometimes cells not develop

3. Local delivery of bioactive molecules

Add bioactive molecule in polymer  released
as polymer degrade  help cell grow and
develop

4. Pre-vascularization

Add blood vessels in polymer  ensure blood

vessels work after implantation
(for nutrient and waste removal)

Sometimes cells develop but blood vessels not

5. In situ polymerization
Liquid scaffold (pre-polymer)  scaffold fit
perfectly without defect

Porogen (solid)  dissolve  cavity  pores

 cells and blood vessel grow and connected

Disadvantage: need to control porosity, must

have full polymerization (can be toxic)
Applications for TE strategies OCTAM
1. Organ shortage
2. Alternative approach to current clinical therapies (i.e. tissue graft and transplants)
3. Treat diseases that are too complex to treat with drugs or genes – can test on organ before human
4. Alternative to animal experiments
5. Market potential – lab grown meat

Organ transplantation RITAAXM

Autologous treatment BIMRITP

Biopsy patient  isolate genes  modify genes  repair cells  inject back  tracking of cells in vivo and
personalized medicine

Key components of TE SCS

1. Scaffold SABM
 Artificial structure to support 3D tissue formation, recapitulating the in vivo environment and allowing
cells to influence their own microenvironment
 Scaffold architecture and fabrication method depends on application (architecture affect cell binding)
 Various biomaterials in numerous architectures serve as scaffolds, providing template for cells to
adhere, interact, proliferate, and restore the function of the impaired tissue
 3D scaffold mimic ECM: growth, migration, differentiation, proliferation, apoptosis, adhesion

Extracellular Matrix (ECM) ACS CDI

 Acellular material around cells, main component: collagen, cell grow ECM to replace scaffold
 Role
 Cell attachment
 Direct cell differentiation
 Induce constructive host tissue remodeling responses

Functions of scaffold TDMS

 Temporary physical support – development of tissues of specific shapes
 Delivery vehicle – cells, growth factors, and genes
 Matrix for cell adhesion – facilitate/regulate cellular processes
 Structurally reinforce defect – maintain defect shape and prevent distortion of surrounding tissue

Requirements of scaffold BBSMPP

 Biodegradable/bioresorbable – controllable degradation and resorption rate
 Biocompatible – should not evoke acute inflammation in vivo
 Suitable surface chemistry – cell attachment, proliferation and differentiation
 Mechanical properties – match those tissue at the site of implantation
 Easily processed – form a variety of shapes and sizes
 Porous (interconnected network) – for cell/tissue growth, nutrient and metabolic waste transport

Balancing various factors PD

 Porosity
 Higher: more and faster tissue and blood vessel ingrowth (+)
 Higher: increases risk of mechanical failure before new tissue has grown (-)
 Degradation
 Too slow: prevents new tissue growth (-)
 Too fast: fracture of material before the new tissue has formed (-)

Important properties of scaffolds PSSD

 Pore architecture NBT
 Allow nutrients to permeate
 Blood vessel infiltration
 Tissue integration
 Surface properties BIG
 Biocompatible
 Integration with host environment
 Encourage cells to grow
 Stiffness/bulk properties CD
 Affects cellular processes
 Degradation profile
 Delivery of scaffolds in vivo IS
 Injectable scaffolds
 Self-assembled scaffolds

Scaffold properties – pore architecture PPAISS

 Permeability (m4/N-s)
 Porosity (%)
 Surface area (mm2)
 Pore interconnectivity (%)
 Pore shape
 Pore size (mm) ~ 1-10 x 10-6 m

Function of pore NBCD

 Transport of nutrient
 Blood vessel progression
 Cell attachment
 Reduce degradation (faster/slower ????)

Scaffold properties – degradation SFT

 Advantages SAC
  No second surgery for removal
 Avoid stress shielding (when implant much stronger, tissue around become weaker because no
need to be that strong)
 Tremendous potential as the basis for controlled drug delivery
 Need scaffold to lose strength as new tissue gains strength
 Need to factor that some people heal slower (e.g. diabetics, smokers, elderly)
 Total material need to maintain strength to a certain level

Biodegradable materials GB BS CPEB

 Gradual breakdown of material mediated by specific
biological activity affected by microenvironmental properties
 General types of degradation (polymer)
 Bulk degradation (for TE)
 Surface degradation
 Mechanisms of degradation
 Chemical: e.g. hydrolysis
 Physical: e.g. thermal
 Enzymatic
 Bacteria

Scaffold properties – injectable scaffolds ICSH DISC SCC

 Minimally invasive nature of delivery (i.e. needle), avoid complex surgical procedures
 Possible to co-inject a cell suspension
 Able to take the shape of the tissue defect (in situ formation), more homogeneous distribution of
bioactive molecules
 Minimizes
 Patient discomfort
 Risk of infection
 Scar formation
 Cost of treatment
 Application
 Intrathecal space in spinal cord
 Cell encapsulation
 Cartilage regeneration

Scaffold properties – surface material interaction PSSCD

 First interaction: protein
 Surface property, shape, chemical characteristic of scaffold affect cell differentiation

2. Cells
Cell Sources PCS
 Primary cells NCEH
 Not immortal, limited lifespan (40-60 division before apoptosis)
 Divide  if mistake  undesired (complication)  apoptosis
 Divide  telomere shorter (there is size limit)  apoptosis
 Cultured directly from subject/tissue sample
 Useful for dividing cells (e.g. endothelial, epithelial, smooth
muscle cells)  easy to cultivate
 Not all types are cultivatable (e.g. brain cells)  hard to cultivate
 Cell lines IGRM
 Immortalised cells, able to proliferate indefinitely
 Avoid mistake of replication  immortal
  No reduction in telomere size  no apoptosis
 Generated through mutation or deliberate genetic modification
 Established as representative of particular cell types
 Useful as model cell but no clinical applicability (for research, not for therapeutic)
 Stem cells IDSUP
 Immortal
 Continuous division and differentiation (develop) into various other kinds of cells/tissues
 Self-renewal: numerous cell division cycles while maintaining the undifferentiated state
 Unspecialised function: cells do not perform any physiological function
 Potency: ability to differentiate into specialized cell types under controlled physiological condition
(after differentiation  becomes primary cells)

Sources of stem cells EAI

 Embryronic stem cells – harvested from inner cell mass of the
blastocyst 7-10 days after fertilization
 Pluripotent: can develop for scaffold for any kind of cells
 Ethical problem: must kill to get cell
 Adult stem cells – many adult tissues contain stem cells that can be isolated
 Multipotent: can develop to specialized cell types present in specific tissue/organ
e.g. bone marrow  blood cell, adipocytes  heart, skin cells
 Induced pluripotent stem cells (IPSC) – primary cells that are reprogrammed genetically to go back to
the embryonic state (stemness)

Autologous treatment ??

Use of stem cells TODGC

 Replace tissues/organs
 Repair of defective cell types
 Delivery of genetic therapies
 Delivery chemotherapeutic agents

3. Environment STG BDA

Signalling and bioreactors
 Signalling molecules
 Responsible for transmitting information between cells in the body, important for growth
 Size, shape, target and function vary greatly
 Bioreactors
 Any manufactured or engineered device or system that supports a biologically active environment
 e.g. petridish or something where cell will grow

Types of signals MBP

 Mechanical signals
 Stiffness of material (shear/compression/stretch)
 Biological signals
 Growth factors (biomolecules)
 Physical signals
 Electrical signals (patterns/shapes)

Challenges in TE
HLV CSIT MSR FM CET
 Host integration
 Long-term viability
 Vascularization
 Cells
 Scale-up
 Immune rejection/safety concern
 Tracking of cells in vivo
 Materials
 Materials/scaffolds that more closely replicate complex tissue architecture
 Factors
 Lack of understanding in modulation of cell function in vivo

 Right combination of cell, scaffold, and factors depends on clinical problem

– extensive physician/scientist/engineering collaboration is vital to success
 Tissue engineering is leveraging our knowledge of cell biology and materials
science to promote tissue regeneration where the natural process is not
enough (e.g. stem cells)

TE success and products SBUD CK

 Skin
 Treat burns, ulcers, deep wounds, etc
 Apligraf – dual layer skin equivalent to keratinocytes and fibroblasts on collagen gel
 Epicel – autologous cells grown to cover a wound
 Dermagraft – dermal fibroblast on resorbable polymer
 Cartilage K
 Treats and repairs the cartilage damage in knee
 Carticel – autologous chondrocyte implantation

L05 – Application 2: Drug Delivery

Issues for a drug to reach market APT
 Absorption – go inside
 Premature degradation – after go in
 Toxicity – make sure go to the right cell

Ideal quality of drug product ERS

 Optimum effective – clinical effectiveness, generic effectiveness (blood levels, elimination,
pharmacological response, bioavailability)
 Maximum reliable – stability (chemical, physical, microbiological), unit-dose precision, patient
acceptance (convenience), bioavailability (percentage, uniformity)
 Maximum safety – therapeutic index, side effects (onset, frequency, severity, reversibility), drug
interactions (probability, severity), stability

Drug Delivery STELED

 Control the spatial and temporal concentration of drug in the body to maximize its effect
 More effective, last longer and produce less side effects than tablet/ injection. Deliver drug at a rate
determined by needs of the body over a specified period of time

Essential component of drug delivery Dr DiDiDoDe

 Drug
 Disease
 Disease site
 Dosage (form, physical state of drug)
 Delivery system (drug release mechanism)
Developing DDS DRSAT
 Drug (charge, size, stability)
 Rate of release
 Target site (barriers)
 Route of administration
 Time scale of release

Ideal drug delivery system (DDS) BCCDEFIMSS

 Biocompatible
 Comfortable for patient
 Capable of high drug loading
 Degradable
 Easy to fabricate and sterilize
 Free of leachable impurities
 Inert
 Mechanically strong
 Safe from accidental release
 Simple to administer

Types of commonly used forms TESTISOA

 Tablets, capsules
 Emulsions
 Solutions, syrups, elixirs
 Transdermal patches
 Inserts, implants, devices
 Suspensions
 Ointments
 Aerosol products

Different physiological barriers FRAUT

Body: biological barrier
 Different formulation
 Different route of administration
 Specific accumulation at target site
 Specific uptake in diseased cells
 Specific target of intercellular organelles
from endolysosome/lysosome

Drug delivery routes OTII

 Small molecules: oral, transdermal, injection
 Big molecules (peptide/protein): injection (subcutaneous or intravenous)
 Oral: SQW
Small drugs
Quick onset
Drug wastage

Transdermal: SSL
Small drugs
Slow onset (1-7d, skin regen)
Low dose

Implants: BIS
Big drugs
Invasive
2 surgical procedures

Injection: BIQ
Big drugs
Invasive
Quickest onset

Targeted w/ nanoparticles: IC
IV, mostly for cancer

Conventional delivery via tablets/capsules TSLMA

Tablet  released in stomach (acid) 
liver (detox)  metabolized
(excreted)/absorbed (bloodstream)
 Low bioavailability due to gastro
side effects and liver toxicity

Transdermal, implant, injection bypass

liver

Bioavailability BAE
% of drug that reach bloodstream instead of excreted
Affected by absorption rate and elimination rate

IV
High bioavailability (100% at beginning), slowly eliminated
from blood

Oral
Need more time to reach blood, delayed by stomach and
liver

OSAISITI
8
different ways to administer drugs
1. Peros – oral
2. Peros – sub-linguale (under tongue)
3. Rectale (anal)
4. Intra-venous (directly to blood vessel)
5. Sub cutaneous (below skin)
6. Intradermal (inside skin)
7. Transdermal (between 2 layer of skin)
8. Inhalation

Controlled release: the role of materials CAPF

 Controlling element to retard the release of drug degrade slowly
 Means of attachment of dosage form to epithelium
 Protects/stabilizes drug
 Allows for flexibility in design and performance slowly diffuse

Mechanism of drug delivery CCST

 Conventional (short/long-term) RT
 Require repetitive dosing (~hours) ensure within window
 e.g. conventional delivery via tablet/capsule
>max: toxic
<min: ineffective
Therapeutic index: MTC/MEC

 Controlled rate (long-term) NPAECLB

 Constant concentration over a period of time, no
repetitive dosing (~days), always within window
 Potent drug that are fast degrading (short half-life)
 Avoid under/overdosing
 Reduce side effect
 Better patient compliance
 Can bypass liver
 Low therapeutic drugs e.g. blood-pressure lowering
drugs

 Sustained (long-term) CLHCPM

 Drug for a chronic condition, release over a longer period
o
 e.g. HGH for dwarfism, prostate cancer treatment, female
menopause, contraceptive

 Targeted TILCHR
 Only target cells that need therapy or repair
 Usually implanted/injected
 Localized release (close to target)
 e.g. cancer, heart cells, retina

Release duration PPI

Depends on the formulation and application
 Procardia XL (pill) – 24 hours
 Patch – weeks
 Norplant (implant) – 5 years

Factors influencing release profile DME

 Drug physiochemical properties SDAH
 Molecule size, diffusivity
 Hydrophilicity
 Molecular interaction with other substances
 Affinity to bulk material
 Half-life/drug stability
 Material physiochemical properties DMPA
 Degradation rate
 Mn/Mw (for polymers)
 Porosity
 Pores distribution and interconnectivity
 Surface area
 Hydrophilicity of the material
 Environment FPTE
 Fluid flow
 pH and temperature
 Enzymes, proteases, and other cell secretion products
 Any other factor that may affect degradation of material and drug concentration in the medium

Controlled DDS release (tutorial) DWC

 Diffusion controlled MRMM
 Membrane-reservoir
Drug dissolved/dispersed in reservoir
surrounded by rate limiting inert membrane

 Matrix-monolithic
Drug dissolved/dispersed in matrix. Release rate
of drug from matrix is not constant (decreasing)
Factor: drug size, drug type, matrix nature

 Water-penetration controlled OS
 Osmotic pumps ODPSP EPCM
 Containing osmogen and drug, separated by piston that will push drug out the orifice as the
osmogen absorb water. Outer layer coated with semipermeable membrane. Polymer matrix for
control
 Application: Oral and implants
 Elementary osmotic pump (EOP)
 Push pull osmotic pump
 Controlled porosity osmotic pump
 Multiple drug-releasing osmotic pump
 Swelling controlled WSAM
 In the body, absorb water  swell  increase in aqueous content in the formulation 
increase in mesh size of the polymeric network  drug diffuse to external environment
 Chemical controlled BE
 Biodegradable/erodible
 Drug dispersed in polymer matrix unable to diffuse out immediately. As polymer degrade 
drug released. No need removal
 Bulk degradation
 Surface degradation
Diffusion vs Osmosis

Taking advantage of pathological condition to design efficient delivery system

Stimuli responsive
nanoparticles IE

 Internal RPTE

 Redox status
 pH
 Temperature
 Enzymes

 External LUM
 Light
 Ultrasound
 Magnetic field
Tumor/inflammation-mediated overexpression of proteins/enzymes (by Enzyme) TECOP
Tumor  specific enzyme produced
 contact with particle  cut 
open  particle drug

Immuno nanoparticles ILACR

Immunoliposomes: liposomes with an antibody attached to
their surface

Strategy for cancer therapy: conjugation of liposome surface

with anti HER2 Ab, anti EGFR (epidermal growth factor)
antibody

Put receptor  target and enter cell through

immunonanoparticle

Enhanced temperature (by Heat) IE LH

2 source:
 Internal stimulus – inflammation causes an
increase of T at disease site
 External stimulus – from outside body by
ultrasound or high-frequency AMF
(oscillation and heat release)

e.g.
temperature-sensitive lipidics nanoparticle + hyperthermia produced by ultrasound
Lipid have TM 39-42C, >42 will become leaky (destabilize layer) and release cargo (drug)
Drug released only at disease are (high T)

pH partition in normal and cancer cells (by pH) DPDR

Different internal pH in cancer cells  particle
go in  pH degrade particle and release drug

Nanomedicine 1D 7-200nm
1 dimension in nano scale  nanomedicine, but 7-200nm more effective

Nanomedicine application REACTL

 Reach sites not accessible to bigger carriers
 Evasion from entrapment by Mononuclear Phagocyte System (MPS) for particle >200nm
 Avoid premature elimination through kidney for particle <7nm
 Able to carry hydrophobic and/or hydrophilic drugs
 Functionalize surface for targeting
 Localized release: release drug in specific area that are affected (increase pharma effect)

DDS Nanoparticle ??
 A) Mimic the membrane of cell to fool body
Green (core): hydrophilic drug
Red (lipid): hydrophobic
Green (surface): contrast agent
Black (surface): antibody for targeting
Black (PEG): longer circulation time

B) Solid biodegradable polymer

Green (core): hydrophilic drug
Red (core): hydrophobic drug
Core: Biodegradable for drug release

C) Hydrophobic dendrimer
Red (in) hydrophobic

Passive targeting: no receptor, just accumulate

nanoparticle passively inside the target cell

Active targeting: attach receptor on particle surface for 1

type of cell  targets and recognize specific receptor (e.g.
cancer)
Extravasation through leaky vasculature – enhanced permeability and retention (EPR) RNTP
Red particle: nanoparticle
Normal tissue: endothelial lining intact, red in
blood
Tumor tissue: endothelial lining disrupted, red
penetrate tissue
If too big, cannot pass
Passive targeting: there is space where red
accumulate inside tumor, must have long
circulation time to allow more accumulation

FGL
Free drug: go in and eliminated by kidney
Giant liposome: bigger, a bit better
Liposome: low elimination, good circulation

PEGlated liposomes CBRD

 Prolong circulation  more
accumulation
 Reduce blood protein adsorption
 Decrease recognition by RES
(reticuloendothelial system)
 Downside: decrease cellular uptake
HWSPCA
Hydrophilic PEG  water shell  surrounds liposome surface and forms a steric barrier  protect
liposome from RES cells, serum proteins, and self-aggression  longer circulation  more accumulation
for passive targeting

A) Better than normal drug: can have +/- charge lipid,

hydrophobic/philic
B) +PEG  longer circulation
C) +PEG +targeting  better circulation, active &
passive
D) +ligand +imaging agent  agent follow particle to
see where is it

Application of liposomes COPOT

 Cancer chemotherapy – entrap anticancer drug: increase circulation time, protects from metabolic
degradation
 Carrier of drug in oral treatment – steroids used for arthritis incorporated into large MLVs. Oral
administration of liposome encapsulated insulin in diabetic animal.
 Pulmonary delivery – inhalation devices (nebulizers) to produce aerosol droplets containing liposomes
 Treatment of drug overdose – particulate carriers act as sink, reduce the bioavailable drug
 Topical applications – drugs like triamcilone, methotrexate, benzocaine, cortisteroids, etc. successfully
incorporated as topical liposome

Liposomes for vaccine application

 Normal vaccine: inject dead virus 
body recognize but virus cannot
reproduce  body produce antibody
against that virus  protection

This one: covid virus identified by spike

protein  synthetic mRNA sequence
packed in lipid nanoparticle  delivers
instruction to cell  produce spike
protein  develop antibody

Vaccine usually intravascular/dermal

Doxorubicin
 Potent anticancer drug with high cardiotoxicity due to non-specific accumulation in cardiomyocytes
 Liposomal doxorubicin: no cardiotoxicity from doxorubicin, but associated with predictable muco-
cutaneous toxicity (hand-foot syndrome) due to preferential accumulation of DOX in the skin. Short
term effect triggered by DOX
 Up to 10% patient experience complement activation-related pseudoallergy (CARPA) effect due to lipid
bilayer (not the drug)
Unsolved challenges:
 CARPA
 Toxic accumulation of NPs
CARPA and NP due to use of synthetic nanoparticles, recognized by body as non-endogeneous material
 Poor correlation between in vitro and in vivo experiments
 Lack of standardization and lack of proper regulations at the nanoscale
 Why not use patients own cell??

The ideal nano-carrier – necessary to be multifunctional?

 Put all function  not ideal
Too complex will not work, sometimes simple ones work better than complex
Different factors, not only function

Nature, size, structure  affect circulation half-life, extravasation, penetration, internalization, drug
release kinetics  impact therapeutic efficacy and durability