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TOXICITY MODEL

Zebrafish is a useful tool of vertebrate model to determinate and investigate the toxicity (safety) of targeted compounds on different stages. Safety pharmacology identify potential adverse effects in drug candidates early in the drug development pipeline, contributing to reduces the overall cost of the drug-discovery phase. Faster, cheaper and more reliable assays based on zebrafish are being developed as major tools for assessing toxicity of chemicals during the drug-discovery process.

Acute Toxicity

The aim of the acute toxicity is to determine the toxicity of targeted compounds on embryonic stages of zebrafish.
The test uses the lethal dose 50 (LD50) to determine the potential toxicity of the compounds.

Endpoints
  • NOEC
  • LOEC
  • LD50
  • Apical parameters
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Acute toxicity scheme

Chronic Toxicity

The aim is to study the long-term effect of target compound in zebrafish embryos.

Endpoints
  • NOEC
  • LOEC
  • Different effect concentration
  • Sublethal effects
  • Reproduction and growth effects
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Chronic toxicity scheme

Organ-specific Toxicity Assays

Cardiotoxicity

The aim is to assess the effect of target compounds on zebrafish embryos, evaluating cardiac activity.

Endpoints
  • ECG Patterns
  • Beats per minute /second
  • Blood flow activity %
  • Biomarkers (RT-PCR)
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Cardiotoxicity scheme

Hepatoxicity

The aim is to assess liver damage of a target compound in zebrafish embryos.

Endpoints
  • Size modification
  • Necrosis
  • Degeneration
  • Molecular analysis (RT-PCR)
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Hepatoxicity scheme

Neurotoxicity

The aim is to assess neurotoxicity of target compound using zebrafish embryos.

Endpoints
  • Burst activity
  • Embryo inactivity
  • Molecular analysis (RT-PCR)
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Neurotoxicity scheme

IBD Model

Inflammatory bowel diseases (IBD) constitute complex and multifactorial diseases difficult to replicate in vitro. The use of in vivo models can resolve this limitation. In the recent years Zebrafish in vivo models of chemically-induced enterocolitis have arisen as an attractive alternative for the study of the molecular mechanism of the disease as well as the screening of new treatments.

Embryo Model

The aim is to develop a useful tool to screen bioactive drugs or food supplements that modulate inflammation.

Endpoints

Functional analysis

  • Study of the changes in the absorptive intestinal region
  • Study of the peristaltic movements

Molecular Analysis

  • Expression levels of pro/anti-inflammatory, proliferation and differentiation markers

Histological Analysis

  • Changes in the intestinal architecture
  • Study of differentiated cell types (goblet cells…)
  • Neutrophil and macrophage infiltration

Infectious Models

Ikan Biotech can generate preclinical infectious models with:

Gram Positive

  • Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus.

Gram Negative

  • Acinetobacter baumannii, Haemophilus spp., E. coli, Vibrio parahemolyticus. 

Other

  • Mycobacterium marinum, Candida albicans.

The aim of this model is to reproduce infectious diseases in zebrafish to test efficacy of new treatments or vaccination in early stages.

Endpoints

In vitro

  • Minimun inhibitory concentration (MIC)

In Vivo

  • Treatment
  • Vaccination
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Treatment or vaccination scheme for infectious deseases

Tumor Models

A major difficulty found in effective cancer treatment is the complexity of the biological mechanism underlying cancer onset and disease progression. This is because the genetic makeup and metabolic profile of each individual patient influence the effect of anticancer drugs. As such, different people respond differently to the same therapy: while one treatment brings about the desired success in one group of patients, it does not change the condition of other groups at all and may even leads to adverse effects.

Ikan Biotech has designed an innovated strategy for prevention and treatment of cancer. The chance to carry out a dynamic display of the cancerous process in vivo offers us the unique opportunity to understand this subjacent biology and design a tailor-made treatment for each case.

Xenograft

Microinjection of tumor cells in zebrafish offers an efficient tumor model system to design and test the efficacy of potential drugs for the treatment of cancer. This model offers a wide variety of assays that are suitable for personalized therapeutics design.

Scheme of Xenograft

Supporting assays

3D SPHEROIDS ASSAY

3D Spheroids assay is a powerful technique to assess the biological effects of cell proliferation, migration and invasion system.

CYTOTOXICITY ASSAY

Cytotoxicity leads one of the characteristic step in drug discovery. Ikan Biotech provides cytotoxicity assay to asses drug safety profiles.

CELL MIGRATION CAPACITIES

In vitro scratch assay is a well-known method to measure cell migration. This study is suitable for the evaluation on the drug effect on cell migration.

GENE EXPRESSION ASSAY

The determination of the pattern of genes expressed provides specific information about cellular function. Gene expression profiling allows to assess the effects of anti-tumor drugs by the determining which genes are expressed.

Others

Obesity model

According the The World Health Organization (WHO) raised body mass index (BMI) is a major risk factor for noncommunicable diseases such as:

  • Cardiovascular diseases (mainly heart disease and stroke)
  • Diabetes
  • Musculoskeletal disorders (especially osteoarthritis)
  • Some cancers (including endometrial, breast, ovarian, prostate, liver, gallbladder, kidney, and colon)

Above indicated disease are health consequences of overweight and obesity.

Ikan Biotech has established an obesity model in zebrafish allowing us to go in further detail in the study of the molecular mechanism of the disease, as well as the screening of new treatments and to include in a defined study and additional obese group to compare the effects of given compound with a normal weight group.

Adult Model

The aim is to offer a tool for studying different food supplements, diets as well as analyzing how the candidate compounds affect regular and obese population.

Endpoints

Efficacy analysis

  • Efficacy study comparing the effects of a compound in a normal weight population vs obese population.

Molecular Analysis

  • Expression levels of target genes and markers

Histological Analysis

  • Adipocytes size
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Tail Fin Regeneration Model

In mammals, such as mice, humans and other primates, the capacity of regenerate tissues after an amputation is very reduced, in fact, the digit tips are the only part of the limbs that can be regenerated.

The aim of this assay is to study the capacity of the target compound to enhance tissues regeneration in zebrafish.

Endpoints

Efficacy analysis

  • Determination of the outgrowth length from the amputation plane, length measurements and time-lapse imaging.

Molecular Analysis

  • Expression levels of target genes and markers.
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Regeneration Model