dabur

Est.1979

Providing Integrated Research Solutions in Preclinical Biology

Modules

In Vitro Pharmacology

Inflammation

in vitro Model for identification of anti-inflammatories
ModelChemokine & cytokine profiling of potential anti inflammatory molecules using Dendritic cells
Test system
  • Murine bone marrow derived dendritic cell cultures
Methods
  • Generation of Dendritic cells in vitro
  • Treatment of DCs with LPS and test compounds
  • Effect on secreted levels of key pro-inflammatory markers
End points
  • Quantitation of TNF-a, IL-1-ß, IL-6, IL-8, MIP-1-a, RANTES, CCL-22, IP-10

in vitro Model for identification of anti-arthritic compounds
Modelin vitro Model for identification of anti-arthritic compounds
Test system
  • Human synovial- sarcoma cell line (SW982)
  • Human chondro-sarcoma cell line (SW1353)
Methods
  • Treatment of cells with test compounds
  • Effect on cellular proliferation Effect on cytokines/chemokines activity that play important role in pathogenesis of arthritis) secreted by LPS/TNF-a/IL-1- stimulated cells – -TNF- a, IL-1- , IL-6, MIP-1- a and MMPs.
End pointsAnti-inflammatory effect on secretion of cytokines and matrix metalloproteinases

Model for Xanthine Oxidase inhibitory activity – management of Gout
ModelEvaluation of anti-inflammatory activity in respiratory diseases such as asthma
Test system
  • Human lung epithelial cell line (A549)
  • Bronchial Smooth Muscle Cell line (BSES-2B)
Methods
  • Treatment of cells with test compounds
  • Effect on the secretion of the inflammatory mediators secreted by TNF-a stimulated cells treated with test compounds : IL-6, IL-8,IL-1ß, MCP-1, TNF-a, RANTES and GMCSF
End pointsQuantitation of inflammatory cytokines

 

in-vitro Model for protective activity against Inflammatory Bowel Disease (IBD)
Modelin-vitro Model for protective activity against Inflammatory Bowel Disease (IBD)
Test system
  • Human intestinal epithelial cells (Caco-2)
  • Human colonic epithelial cell line (HT-29)
Methods
  • Identification of non-cytotoxic concentrations
  • Treatment of cells with test compound
  • Stimulation of cell line with appropriate stimulus (TNF-a or IL-1-ß)
  • Effect on pro-inflammatory markers in culture supernatants by ELISA – IL-8, IL-1ß, IP-10 and MIG
End pointsInhibitory effect on secretion of pro-inflammatory cytokines from intestinal and colonic epithelial cells.

In Vitro Model for identification of anti-inflammatories
ModelChemokine & cytokine profiling of potential anti inflammatory molecules using Immune cells
Test system
  • Murine macrophage cell line (RAW264.7)
  • Murine splenocytes
  • Human monocytic cell line (THP-1)
Methods
  • Treatment of cells with LPS and test compounds
  • Effect on secreted levels of key pro-inflammatory markers
End pointsQuantitation of TNF-a, IL-1-ß, IL-6, IL-8, MIP-1-a, RANTES, CCL-22, IP-10

 

Model for identification of anti-inflammatories for respiratory inflammation
ModelEvaluation of anti-inflammatory activity in respiratory diseases such as asthma
Test system
  • Human lung epithelial cell line (A549)
  •  Bronchial Smooth Muscle Cell line (BSES-2B)
Methods
  • Treatment of cells with test compounds
  • Effect on the secretion of the inflammatory mediators secreted by TNF-a stimulated cells treated with test compounds : IL-6, IL-8,IL-1ß, MCP-1, TNF-a, RANTES and GMCSF
End pointsQuantitation of inflammatory cytokines

 

in vitro Model for screening ocular inflammation
ModelEvaluation of anti-inflammatory activity in ocular inflammation
Test system
  • Conjunctival epithelial cell
End Point
Effect on the secretion of the inflammatory mediators; secreted by TNF-a stimulated cells treated with test compounds – IL-12p40, GM-CSF, MCP-1, IL-6, IL-7, IL-8, IFN-?, IL-1a, IL-1ß, IL-3, and IP-10 and RANTES
in vitro Model for screening ocular inflammation
ModelEvaluation of inflammatory/anti-inflammatory activity in dermal cells
Test system
  • Human skin fibroblast cell line
End points
  • Morphology of fibroblasts
  • Effect on levels of key pro-inflammatory markers secreted by PMA (phobol-12-myristate-13-acetate)-stimulated skin fibroblasts treated with test compounds–GM-CSF, IL-1-b
  • Effect on Nitric Oxide levels
  • Effect on levels of COX, Prostaglandins and Arachidonic acid

 

in vitro Model for screening ocular inflammation
ModelEvaluation of inflammatory/anti-inflammatory activity in dermal cells
Test system
  • Human skin fibroblast cell line
End points
  • Morphology of fibroblasts
  • Effect on levels of key pro-inflammatory markers secreted by PMA (phobol-12-myristate-13-acetate)-stimulated skin fibroblasts treated with test compounds–GM-CSF, IL-1-b
  • Effect on Nitric Oxide levels
  • Effect on levels of COX, Prostaglandins and Arachidonic acid
Evaluation of analgesic/antinociceptive activity
ModelEvaluation of analgesic/antinociceptive activity
Test system
  • Human neuroblastoma/glioblastoma cell lines expressing µ and dopioid receptors Induction of pain response by VIP/Forskolin
End Points
  • Effecton following markers secreted by Forskolin/IL-1-ß stimulated cell streated with test compounds:
  • Effect on PGE2 synthesis
  • Effect on COX-2 activity
  • Effect on cAMP levels
  • Effect on calcium signaling pathway
ModelEvaluation of inflammatory/anti-inflammatory activity
Test system
  • Murine Macrophage cell line (RAW264.7)
  • Human Monocyte cell line (THP-1)
End Points
Morphology of macrophages/monocytes Effect on levels of key pro-inflammatory markers secreted by LPS-stimulated cells treated with test compounds – TNF-., IL-1-., MIP-1-,, IL-6, RANTES, IL-12, IFN-., CCL-22, IP-10, IL-4, IL-10, TGF-b Effect on Nitric Oxide levels
Test systemMouse splenocytes
Stimulatory agent for IL-17secretion
Concanavalin A (5 µg/ml)
Experimental Designe
  • Splenocytes were treated with test item at non-cytotoxic concentrations (1:100 to 1:5) along-with the inflammatory stimulus (ConA-5 µg/ml)
  • After 24 h, culture supernatants were collected from each well
  • Secreted levels of IL-17 were estimated by ELISA
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