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
in Vitro Model for identification of Immunomodulators
Model Chemokine & cytokine profiling for identification of immunomodulators
Test system Murine bone marrow derived dendritic cell cultures
Method
  • Generation of Dendritic cells in vitro
  • Characterization of dendritic cells
  • Profiling of potential of test item for modulation of cytokines & chemokines
  • Validation with immunosuppressant & immunostimulatory molecules
End points Quantitation of TNF- a, IL-1- ß, MIP-1- a , IL-6 ,RANTES, IL -12, IFN– ?, CCL- 22, IP -10, IL -4, IL -10 & TGF-ß
X
  • Skin health
    Screening models for assessment of skin rejuvenation potential
    Model In vitro cell based models
    Test System Human 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
    X
  • Skin senescence
    Testing of Products with anti ageing & anti wrinkling potential
    Model In vitro cell based models
    Test system Human 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
    X
  • Skin lightening / depigmentation
    Testing of Products with anti ageing & anti wrinkling potential
    Model In vitro cell based models
    Test system Human 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
    Model In vitro cell based models
    Test system Fibroblast 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 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 system Human 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
    X
  • Skin Whitening
    Testing of Products with potential for skin whitening
    Model In vitro cell based models
    Test system Murine 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
    X
  • Anti-blemish
    Testing of Products with potential to heal skin blemishes
    Model In 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
    X
  • Skin Photoaging
    Screening models for assessment of anti photoageing potential in vivo
    Model In vitro models
    Test system Nude 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
    X
  • Chemokine & cytokine
    Chemokine & cytokine profiling of potential anti inflammatory molecules
    Model in 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
    X
  • Screening permeability Skin
    Screening models for assessment of permeability through skin
    Model In vitro model
    Test system Franz 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
    X
  • Dermal Anti – oxidants
    Screening models for assessment of anti-oxidant potential
    Model in vitro cell based models
    Test system Human 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
    X
  • Hair growth promoter
    Screening model for hair growth promoter
    Model In vitro model for hair growth promotion
    Background The 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-)
    X
  • Skin barrier / permeability assessment
    Screening models for assessment of permeability through skin
    Model In vitro model
    Test system Franz 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
    X
  • Dermal Anti – inflammatories
    Chemokine & cytokine profiling of potential anti inflammatory molecules
    Model in 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
    X
Invitro Dermapathology Screen
in vitro Model for identification of Immunomodulators
Model Phagocytosis assay for identification of immunostimulants
Test system Macrophage cell line (RAW 264.7)
Method
  • Treatment of macrophages with test compounds
  • Evaluation of immunostimulatory effect of test compounds exhibited by internalization of particulate material (Zymosan) by macrophage cells
End points Colorimetric estimation of Zymosan uptake by cells
X
  • Psoriatic
    Screening of anti-psoriatic compounds
    Model HaCaT (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
    X
  • Warts
    Screening of compounds for warts
    Model Evaluation 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.
    X
  • Acne
    Testing of Products with potential for treatment of acne
    Model In 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
    Model In 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
    X
  • Leucoderma/Anti vitiligo
    Anti-vitiligo (leucoderma)/ repigmentation
    Model In 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
    X
  • Inflammation
    Testing of Products with potential to reduce inflammation in skin
    Model In 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
    X
  • Wound Healing
    Screening models for assessment of wound healing potential in vitro
    Model In 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
    X
  • Leucoderma
    Screening models for assessment of potential of products for repigmentation
    Model In vitro cell based models
    Test system Primary 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
    X
  • Antiacne compound
    Screening of Antiacne compound
    Model In 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
    X