Chapter
1 - Development of In Vitro Toxicology: A Historic Story
WHAT IN VITRO TOXICOLOGY AND HOW IT CAME INTO BEING?
WHY IN VITRO NOT IN VIVO?
THE REGULATORY CONTROL AND POLICY MAKERS
PROJECTS AND DEVELOPMENTS
SOME OF THE VALIDATED IN VITRO ALTERNATIVES AND METHODS
THE AVAILABLE IN VITRO MODELS SYSTEMS
Stem Cells and Induced Pluripotent Stem Cells
BASAL TO ORGAN-SPECIFIC ENDPOINTS OF IN VITRO TOXICOLOGY
Basal Toxicity or Cytotoxicity
Tetrazolium Bromide Salt Assay
Lactate Dehydrogenase Assay
Acetoxymethyl Diacetylester of Calcein Cytotoxicity Assay
Adenosine Triphosphate Quantification and Adenosine Triphosphate/Adenosine Diphosphate Ratio Assay
CELLS TO 3D ORGANOIDS: IN VITRO ORGAN SYSTEM TOXICOLOGY
SUGGESTED WEB PORTALS FOR FURTHER READING
2 - Principles for In Vitro Toxicology
GUIDELINES GOVERNING THE IN VITRO METHODS FOR TOXICITY TESTING
OPTIMIZATION OF CELL CULTURE
MODELS OF IN VITRO TOXICITY
Three Dimensional Culture Systems
DEVELOPMENT OF BIOMARKERS FOR IN VITRO TOXICITY EVALUATION
ASSAYS FOR SCREENING OF TOXICANTS
The Hen’s Egg Test–Chorio Allantoic Membrane Test
3T3 Neutral Red Uptake Test
Tetrazolium Salt, 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide Assay
The Bovine Cornea Opacity/Permeability Test
DOSE–RESPONSE RELATIONSHIP
Evaluation of DR Relationship
VALIDATION OF IN VITRO TOXICOLOGY TEST METHODS
3 - Models and Methods for In Vitro Toxicity
NEED OF IN VITRO MODELS FOR TOXICITY ASSESSMENT
VALIDATION OF IN VITRO TEST METHODS
Criteria for Regulatory Acceptance
MODELS FOR IN VITRO TOXICITY ASSESSMENT
In Vitro Models for Cytotoxicity Studies
In Vitro Models for Specific Toxicity
In Vitro Models for Liver Toxicity
In Vitro Models for Lung Toxicity
In Vitro Models for Neurotoxicity
In Vitro Models for Immunotoxicity
METHODS EMPLOYED FOR IN VITRO TOXICITY ASSESSMENT
Neutral Red Dye Uptake Assay
Propidium Iodide Uptake Assay
Lactate Dehydrogenase Assay
Trypan Blue Exclusion Assay
Comet (Single-Cell Gel Electrophoresis) Assay
Chromosomal Aberration Assay
Sister Chromatid Exchange Assay
Cell Cycle and DNA Content Analysis
Apoptosis Detection Assay
Forward Gene Mutation Assay (Mammalian Model)
HGPRT Gene Mutation Assay
Mouse Lymphoma Assay (L5178Y TK+/−)
Ames Test (Nonmammalian Model)
Cellular and Functional Responses (Protein/Gene Expression)
STATE OF ART AND RECENT DEVELOPMENT REGARDING IN VITRO APPROACHES
Bioinformatics and Computational Toxicology
Integrated Testing Strategies
CHALLENGES AND CONSIDERATIONS
4 - In Vitro Gene Genotoxicity Test Methods
IMPORTANCE OF GENOTOXICITY TESTING
THE STANDARD TEST BATTERY FOR GENOTOXICITY
GENERAL PROTOCOL FOR IN VITRO TESTS
Bacterial Reverse Mutation Test (TG 471)
In Vitro Mammalian Chromosome Aberration Test (TG 473)
In Vitro Mammalian Cell Gene Mutation Tests Using the Hprt and xprt Genes (TG 476)
In Vitro Mammalian Cell Micronucleus Test (TG 487)
In Vitro Mammalian Cell Gene Mutation Tests Using the TK Gene (TG 490)
CURRENT LIMITATIONS OF IN VITRO GENOTOXICITY TESTING
Faulty Metabolic Activator System
FUTURE PROSPECTIVE OF THE GENOTOXICITY TESTING
5 - In Silico Approaches for Predictive Toxicology
IN SILICO TOXICOLOGY FRAMEWORK FOR TOXICITY/SAFETY PREDICTION
QSAR FOR PREDICTIVE TOXICOLOGY
DESCRIPTORS FOR PREDICTIVE TOXICOLOGY
DATABASES AND WEB TOOLS FOR PREDICTIVE TOXICOLOGY
6 - The Use of Transcriptional Profiling in In Vitro Systems to Determine the Potential Estrogenic Activity of Chemicals of Interest
WHY DO WE HAVE TO ASSESS THE POTENTIAL ESTROGENICITY OF CHEMICALS OF INTEREST?
Transcriptional Profiling in In Vitro Systems to Determine the Potential Estrogenic Activity of Chemicals of Interest
Connectivity Mapping to Define Mode of Action
In vitro to in Vivo Extrapolation
7 - Extrapolation of In Vitro Results to Predict Human Toxicity
NEED TO UNDERSTAND TOXICITY
ORGAN LEVEL AND SYSTEM LEVEL TOXICITY
TOXICITY ASSESSMENT IN VITRO: ADVANTAGES AND DISADVANTAGES
In Vitro Systems for Liver Toxicity Assessment
In Vitro System for Nephrotoxicity Assessment
TO OVERCOME IN VITRO ASSAY LIMITATION
EXTRAPOLATION OF IN VITRO INFORMATION TO IN VIVO FOR HUMAN: SUCCESSFUL AND UNSUCCESSFUL OUTCOMES
UPCOMING POTENTIAL METHODS TO PREDICT TOXICITY
8 - Role of Molecular Chaperone Network in Understanding In Vitro Proteotoxicity
BALANCE BETWEEN PROTEIN FOLDING AND MISFOLDING: MISFOLDED PROTEINS ARE TOXIC SPECIES
MECHANISMS OF CELLULAR PROTEOTOXICITY
CELLULAR DEFENSE AGAINST PROTEOTOXICITY: MOLECULAR CHAPERONES
ROLE IN PROTEOTOXICITY: MOLECULAR CHAPERONES ANTAGONIZES THE ACCUMULATION OF PROTEOTOXIC PROTEINS
THE CELLULAR DEGRADATION MECHANISM IN PROTEOTOXICITY
THE UBIQUITIN PROTEASOME SYSTEM
SPECIFIC ROLE OF DEGRADATION PATHWAYS IN MITIGATING PROTEOTOXICITY
TOXICITY OF AMYLOID BETA IN AD
TOXICITY OF Α-SYNUCLEIN IN PD
TOXICITY OF HUNTINGTIN PROTEIN IN HD
PRION PROTEINS IN ENCEPHALOPATHIES
9 - Scientific and Regulatory Considerations in the Development of in Vitro Techniques for Toxicology
IN VITRO ASSAYS FOR SAFETY AND TOXICITY ASSESSMENT
Use of Cytotoxicity as a Predictor for Acute System Toxicity
Endpoints of Cytotoxicity Studies
Applications of Various Conventional and Evolving Techniques for Evaluating Cellular Response Leading to Toxic Insult
Benefits of Cytotoxicity Testing
Assessment of Carcinogenicity
Tests for Gene Mutation in Bacteria
Tests for Gene Mutation in Mammalian Cells
Tests for Chromosomal Aberration in Mammalian Cells
Nongenotoxic Carcinogenicity Endpoints
Reproductive and Developmental Toxicity Assessment
Tissue Cross-Reactivity Assay
THE APPLICATION OF THE PRINCIPLES OF GLP TO IN VITRO STUDIES
VALIDATION OF IN VITRO METHODS
REGULATORY ACCEPTANCE OF ALTERNATIVE METHODS TO ANIMAL TESTING
TRANSLATIONAL ASPECTS OF IN VITRO ASSAYS
Pitfalls in In Vitro Toxicology
10 - Safety Concerns Using Cell-Based In Vitro Methods for Toxicity Assessment
BIOSAFETY AND RISK MANAGEMENT IN IN VITRO LABORATORIES
RISK ASSOCIATED WITH CELL CULTURE
FACTORS CONTRIBUTING TOWARD RISK IN THE CELL CULTURE
Tissue/Type, Origin, and Source of Cells
Problems Associated With Viral Contaminations
Origin of Viral Contaminations
How Does Mycoplasma Enter Cell Cultures?
TRANSMISSION INVOLVING USE OF NEEDLES AND SHARP OBJECTS DURING MANIPULATIONS
HANDLING CELLS OUTSIDE BIOSAFETY CABINETS
ALTERING CULTURE CONDITIONS
RISK ASSOCIATED WITH PROCEDURES INVOLVING CRYOBANKS/LIQUID NITROGEN
RISK ASSOCIATED WITH PROCEDURES INVOLVING CO2 CYLINDERS
RISK ASSOCIATED WITH PROCEDURES INVOLVING TOXIC COMPOUNDS
PHYSICAL HAZARDS AND OTHERS
CONTAINMENT MEASURES FOR WORKING WITH CELL CULTURES
STATE AND FEDERAL REGULATIONS
Hazard Communication Standards
The Blood Borne Pathogens Standard
The Personal Protective Equipment Standard (29 CFR 1910.132)
The Eye and Face Protection Standard (29 CFR 1910.133)
The Respiratory Protection Standard (29 CFR 1910.134)
The Hand Protection Standard (29 CFR 1910.138)
The Control of Hazardous Energy Standard (29 CFR 1910.47)
11 - In Vitro Methods for Predicting Ocular Irritation
IN VITRO METHODS ADOPTED BY OECD
Isolated Chicken Eye Test
Bovine Corneal Opacity and Permeability Test
Fluorescein Leakage Assay
Reconstructed Human Cornea-Like Epithelium Test Method
RECENT DEVELOPMENT AND LOOKING INTO FUTURE
In Vitro Toxicology
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