dabur

Est.1979

Providing Integrated Research Solutions in Preclinical Biology

Modules

In Vitro Pharmacology

In Vitro Cosmetic Screens

We offer a range of models designed to undertake Dermo – cosmetic assessment of products for a range of indications. These include the following:

In Vitro Cosmetic Screen

Screening models for assessment of skin rejuvenation potential
ModelIn vitro cell based models
Test SystemHuman dermal fibroblast and keratinocyte cell lines
Method
  • Determination of non-cytotoxic concentrations of test item
  • Treatment with non-cytotoxic doses of test item
  • Quantitation of biomarkers for skin health in vitro
End points
  • Fibroblast proliferation
  • Improvement of skin Texture
    — Collagen synthesis
    — Elastin synthesis
    — MMP expression
  • Improvement of Skin moisture
  • Hyaluronic acid synthesis

 

Testing of Products with anti ageing & anti wrinkling potential
ModelIn vitro cell based models
Test systemHuman dermal fibroblast cell line
Method
  • Determination of non-cytotoxic concentrations of test item
  • Treatment with non-cytotoxic doses of test item
  • Challenge with UV-or H2O2 (oxidative stress) to induce premature senescence
  • Comparison of anti ageing potential of test item with reference drugs/ products
End points
  • Cellular viability and proliferation
  • Protection against UV/H2O2 induced damage
  • Levels of extracellular matrix proteins (collagen, elastin, etc.)
  • MMP expression
  • Hyaluronic acid level

Testing of Products with anti ageing & anti wrinkling potential
ModelIn vitro cell based models
Test systemHuman dermal fibroblast cell line
Method
  • Determination of non-cytotoxic concentrations of test item
  • Treatment with non-cytotoxic doses of test item
  • Challenge with UV-or H2O2 (oxidative stress) to induce premature senescence
  • Comparison of anti ageing potential of test item with reference drugs/ products
End points
  • Cellular viability and proliferation
  • Protection against UV/H2O2 induced damage
  • Levels of extracellular matrix proteins (collagen, elastin, etc.)
  • MMP expression
  • Hyaluronic acid level
Testing of Products with anti – tanning potential
ModelIn vitro cell based models
Test systemFibroblast and keratinocyte cells
Method
  • Determination of non-cytotoxic concentrations of test item
  • UV exposure
  • Measurement of cell viability in presence & absence of sun protectors
End points
  • Cell viability as an indicator of UV protection
  • Sun protection factor
  • Protection against UV induced apoptosis
  • Comparison of anti-tanning potential of test item with reference drugs / products
Testing of Products with potential to reduce oxidative stress in skin
Model
  • Cell free assay
  • In vitro cell based models
Test systemHuman dermal fibroblast and keratinocyte cell lines
Method
  • Cell free assay-Free radical scavenging effect by DPPH/ABTS assay
  • Cell based assay
  • Determination of non-cytotoxic concentrations of test item
  • Treatment with non-cytotoxic doses of test item
  • Challenge with UV-or H2O2 to induce oxidative stress
  • Validation with antioxidant molecules or client defined molecules
End points
  • Cell free assay-Free radical scavenging effect
  • Cell based assay
  • Lipid peroxidation
  • Total glutathione levels
  • Super oxide dismutase/catalase enzyme activity
  • Levels of reactive oxygen species
Testing of Products with potential for skin whitening
ModelIn vitro cell based models
Test systemMurine melanoma B16 cells/Primary human melanocytes
Method
  • Determination of non-cytotoxic concentrations of test item
  • Treatment with non-cytotoxic doses of test item
  • Measurement of effect of test item, on melanin turnover
End points
  • Melanin turnover
  • Tyrosinase enzyme activity
  • Comparison of skin whitening effect of test item with reference drugs/ products
Testing of Products with potential to heal skin blemishes
ModelIn vitro cell based models
Test system
  • Human and/or murine fibroblast cell line
  • Murine melanoma B16 cells
Method
  • Fibroblast cell lines-Migration activity
  • Mechanical generation of wound in cell monolayer
  • Treatment with non-cytotoxic doses of test item
  • Dose kinetics/Time Kinetics
  • Melanoma cells- Hypopigmentation activity
  • Melanin turnover
  • Tyrosinase enzyme activity
End points
  • Migration activity in fibroblast cells
  • Hypopigmentation effects in melanoma cells
Screening models for assessment of anti photoageing potential in vivo
ModelIn vitro models
Test systemNude mice
Method
  • Topical application of test formulation
  • UVB exposure for 7 continuous days
End points
  • Visual wrinkling
  • Hydration index with Corneometer
  • Erythema by mexamater
  • Histological analysis
  • Dermal thickness
  • Epidermal hyperplasia
  • Inflammation
  • Collagen estimation
Chemokine & cytokine profiling of potential anti inflammatory molecules
Modelin vitro & ex vivo cell based models
Test system
  • in vitro models
  • Human macrophage cell line ex vivo modelsMurine dendritic cells
Method
  • LPS induced septic shockTreatment of LPS challenged Dendritic/macropghage cells with test item at non – cytotoxic doseValidation with anti – inflammatory molecules or client defined molecules
End points
  • Quantitation of TNF-a, IL-1- a, MIP-1- a, IL-6

 

Screening models for assessment of permeability through skin
ModelIn vitro model
Test systemFranz Diffusion cell Animal skin
Method
  • Skin section application on Franz Diffusion Cell systemApplication of FormulationCollection of receptor solution at defined time pointsProcessing and analysis of receptor solution and skin wash
End points
  • Determination of Drug release/ unit area(ug/cm2)Determination of Permeability coefficient

 

Screening models for assessment of anti-oxidant potential
Modelin vitro cell based models
Test systemHuman dermal fibroblast and keratinocyte cell lines
Method
  • Determination of non-cytotoxic concentrations of test itemTreatment with non-cytotoxic doses of test itemChallenge with UV-or H2O2 to induce oxidative stressValidation with anti – ageing molecules or client defined molecules
End points
  • Lipid peroxidationTotal glutathione levelsSuper oxide dismutase enzyme activityCatalase enzyme activity

Screening model for hair growth promoter
ModelIn vitro model for hair growth promotion
BackgroundThe dermal papilla (DP) is the primary mesenchymal component of hair, which plays pivotal role in hair cycle by induction of new hair follicles and maintenance of hair growth. Keratinocytes also play crucial role in hair growth as the epithelial-counterpart cells. Increase in the proliferation of DPCs and keratinocytes reflect hair growth promoting effect. Stimulation of growth factors, such as VEGF, IGF-1, FGF-7 and inhibition of TGF- secretion also supports hair growth.
Test system
  • Immortalized Human Follicular Dermal Papilla Cells (hHFDPCs)
  • Human normal keratinocyte cell line (HaCaT)
Assays
  • Stimulatory effect on the proliferation of HFDPCs and HaCaT cells
  • Protective effect on cell viability against oxidative stress induced by H2O2
  • Stimulatory effect on secretion of growth factors by HFDPCs
  • Inhibitory effect on TGF- secretion
  • Stimulatory effect on Wnt/ß-catenin Signaling Pathway
End points
  • Proliferation
    – MTT assay/BrdU assay
  • Growth Factors
    – Vascular endothelial growth factor (VEGF)
    – Insulin-like Growth Factor 1 (IGF-1)
    – Fibroblast Growth factor (FGF-7)
    – Transforming growth factor beta (TGF-)

 

Screening models for assessment of permeability through skin
ModelIn vitro model
Test systemFranz Diffusion cell Animal skin
Method
  • Skin section application on Franz Diffusion Cell system
  • Application of Formulation
  • Collection of receptor solution at defined time points
  • Processing and analysis of receptor solution and skin wash
End points
  • Determination of Drug release/ unit area(ug/cm2)
  • Determination of Permeability coefficient

Chemokine & cytokine profiling of potential anti inflammatory molecules
Modelin vitro & ex vivo cell based models
Test system
in vitro models
  • Human macrophage cell line ex vivo models
  • Murine dendritic cells
Method
  • LPS induced septic shock
  • Treatment of LPS challenged Dendritic/macropghage cells with test item at non – cytotoxic dose
  • Validation with anti – inflammatory molecules or client defined molecules
End points
  • Quantitation of TNF-a, IL-1- a, MIP-1- a, IL-6

Invitro Dermapathology Screen

Screening of anti-psoriatic compounds
ModelHaCaT (immortalized human keratinocytes),THP-1 (human monocytic cell line) Human Sebocytes
Test system
  • Curcumin – Antipsoriatic activity
  • Human monocytic cell line (THP-1)
Method
  • Treatment of cells with test compoundsEffect on proliferation of HaCaT/THP-1 cellsEffect on levels of Th-1 cytokines/chemokines secreted by TNF-a stimulated HaCaT/THP-1 cells treated with test compounds –
    – TNF-a, IL-6, IL-8, IFN-?, IL-2, RANTESAssessment of IL-6 & IL-8 in A549 cell lineInduction of apoptosis in HaCaT cells:
    – Annexin-translocation on membrane
    – Mitochondrial potential
    – Caspase-3
    – DNA fragmentation (Hoechst /TUNEL staining)
    – Cell-cycle distribution
End points
  • Anti-proliferative effect
  • Anti-inflammatory activity
  • Pro-apoptotic effect

Screening of compounds for warts
ModelEvaluation of immunostimulatory activity of test molecule useful in the treatment of warts
Test system
  • Human PBMCs / macrophage cell line / Spleen cells
Method
  • Treatment of cells with test molecule. Determination of levels of cytokines (TNF- a, IL-1a, IL-1ß, IL-1RA, IL-6, IL-8, GM-CSF, G-CSF and MIP-1a) Analysis in duplicates.
End points
  • Assessment of cytokine activity (TNF- a, IL-1a, IL-1ß, IL-1RA, IL-6, IL-8, GM-CSF, G-CSF and MIP-1a).Usefulness of test molecule for treatment of warts.

Testing of Products with potential for treatment of acne
ModelIn vitro cell based models
Test system
  • HaCaT(immortalized human keratinocytes)
  • THP-1 (human monocytic cell line) Human Sebocytes
Method
  • Determination of non-cytotoxic concentrations of test item
  • Inflammatory stimulus : P.acnes / LPS
  • Treatment with test items
  • Oil red O staining for lipid synthesis (sebocytes)
  • Anti-inflammatory effect on LPS/P.acnes induced secretion of cytokines; IL-8, IL-6
End points
  • Downregulation of lipid synthesis in sebocytes
  • Inhibition of cytokines which play key role in acne development

Anti-vitiligo (leucoderma)/ repigmentation
ModelIn vitro cell based models
Test system
  • Murine melanoma B16 cells/Primary human melanocytes
Method
  • Melanocyte proliferation assay
  • Melanogenesis stimulation activity
  • Melanin content
  • Tyrosinase activity
  • Melanocyte migration assay
End points
  • Stimulation of melanocyte proliferation
  • Stimulation of melanogenesis
  • Stimulation of melanocyte migration
Testing of Products with potential to reduce inflammation in skin
ModelIn vitro & Ex vivo cell based models
Test system
  • In Vitro models
  • Human dermal fibroblast cells
  • Macrophage cell line
  • Ex Vivo models
  • Human dendritic cells
Method
  • LPS induced septic shock
  • Treatment of LPS induced cells with test item at non – cytotoxic doses
  • Assessment of anti inflammatory potential
End points
  • NO release
  • Quantitation of TNF-a, IL-1- ß, MIP-1- a, IL-6
  • Comparison of anti inflammatory potential of Test item with reference products

Screening models for assessment of wound healing potential in vitro
ModelIn vitro cell based models
Test system
  • Human and/or murine fibroblast cell lines
  • Human dermal keratinocyte cell line
Method
  • Mechanical generation of wound in cell monolayer
  • Treatment with non-cytotoxic doses of test item
  • Dose kinetics
  • Time Kinetic
End points
  • Cellular migration
  • Morphological evaluation of cell spreading by light microscopy
  • Estimation of cytoskeleton proteins
  • MMP
  • Actin

Screening models for assessment of potential of products for repigmentation
ModelIn vitro cell based models
Test systemPrimary human melanocytes and melanoma cells
Method
  • Determination of non-cytotoxic concentrations of test item
  • Treatment with non-cytotoxic doses of test item
  • Determination of the melanogenesis & melanocyte migration activity
  • Validation with anti-vitiligo molecules
End points
  • Cell proliferation
  • Melanin content
  • Tyrosinase activity
  • Cell migration

Screening of Antiacne compound
ModelIn vitro cell based models
Test system
  • HaCaT(immortalized human keratinocytes)
  • HP-1 (human monocytic cell line)
  • Human Sebocytes
Method
  • Determination of non-cytotoxic concentrations of test item
  •  Inflammatory stimulus : Pacnes / LPS
  • Treatment with test items
  • oil red O staining for lipid synthesis (sebocytes)
  • Anti-inflammatory effect on LPS/P.acnes induced secretion of cytokines; IL-8, IL-6
End points
  • Downregulation of lipid synthesis in sebocytes ? Inhibition of cytokines which play key role in acne development

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