This study can be complete up to 14 days’ study, Prophylactic dose can be given up to 7 to 14 days before TNBS induction. Sulfasalazine will be the best as a Standard drug.
How to Induce TNBS in Mice/ By what route TNBS is being induced in Mice?
TNBS induction by PE Tubing (PE9050) and Syringe and needle (24 gauge) can be used. (SDR Scientific), PE tubing can be marked 4 cm, that should be inserted in to the rectum up to 4 cm, sometimes fecal pellets restrict entry in to rectum even after fasting. Animals are given anesthesia thiopental sodium up to 25-40 mg/kg.
How to calculate Disease Activity Index TNBS Induced Colitis Model ?
Disease Activity Index (DAI) can be calculated by scoring Occult blood (using Hemospot Kit- Occult Blood, Occurrence blood in Faces), Fecal Consistency (Normal, Solid, Semisolid, Slurry) and Body weight. All three parameters are recorded from day 0 to day 7 to calculate DAI score.
How to Score the disease level in TNBS Model ?
Mice will be scored from 0 to 3; 0 being normal and 3 being diseased. Moreover, macroscopic lesion score will include length and thickness of the colon, any mucosal lesion. Similar type of lesion score will be given from 0 to 3; 0 being normal and 3 being diseased.
Can be do Microbiota check in TNBS Model ?
Yes, Fecal Collection for microbiota checkup can be done before TNBS induction on day-0, day-3 and at day-7 before dissection. Animals are separated individually to collect fecal pellets. Microbiota are preserved to identify bacterial colony in Faces by q-PCR.
What we should taken care while keeping animal on fasting?
C57bl/6 and BALB/c mice can be use, but as written in many literatures do not keep animals on fasting for 24 hrs., otherwise animal may die or go in coma, if they get in coma, it should be care with gentle for 2-3 days giving 5% dextrose subcutaneously (0.5 ml) three times a day. Mostly avoid animal fasting more than 6 hrs. or keep the animals on dextrose rather than keeping them on fasting. For best results choose SD rats, mice mainly show mortality after TNBS induction.
What activities are performed on day 7 in TNBS study model?
On day 7, blood will be collected from the retro-orbital sinus/ cardiac puncture and serum will be separated for biochemical, cytokine evaluation. Further the animals will be euthanized with overdose of thiopentone (150 mg/kg, administered intraperitoneally) and the colon will be harvested, washed in PBS/ NSS and colon tissue will be promptly stored in -80ºC and fixed in 10% formalin for further cytokine estimation and histological analysis. Colon length, Colon Weight and Colon Diameter are measured on same day.
How to calculate Disease Activity Index (DAI) in TNBS study Model?
Please watch this video to see Disease Activity Index Calculation
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