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A Determination of Tumor Necrosis Factor Expression in TMJ Inflammation with the Use of Microarray Analysis

Department of Biomedical Sciences, Baylor College of Dentistry-The Texas A&M University System Health Science Center, 3302 Gaston Avenue, Dallas, TX 75246, USA;

View larger version (20K): [in a new window]   Figure 1. Effect of CFA treatment on mRNA expression of TNF-, TNF-R1, and TNF-R2 in TMJ tissues. (A) RT-PCR of TMJ tissues demonstrating mRNA levels of TNF-, TNF-R1, and TNF-R2 after treatment with CFA for 48 hrs. ß-actin served as an internal loading control and as a means for normalization. (B) The data are presented as the percent ratios of the densitometric units of the TNF-, TNF-R1, or TNF-R2 mRNA bands to the densitometric units of the ß-actin mRNA band for the CFA-injected animals, then determined as a percent of the uninjected controls. The data are presented as the means ± standard deviations of 3 separate experiments, and significance (*) was determined at a level of p 0.05. (C) Primers and conditions used for PCR examination.

View larger version (29K): [in a new window]   Figure 2. Effect of CFA treatment on protein expression of TNF-, TNF-R1, and TNF-R2 in TMJ tissues. (A) Western blot analysis of TNF- and TNF receptor protein expression in TMJ tissues 48 hrs after CFA treatment. (B) The data are presented as the percent ratios of the densitometric units of the TNF-, TNF-R1, or TNF-R2 protein bands from the CFA-injected tissues to the densitometric units of the ß-actin protein band, then expressed as a percent of the uninjected controls. Data from 3 independent blots, run in triplicate, were statistically analyzed and are presented as means ± standard deviations, and significance (*) was determined at a level of p 0.05 by means of Student’s t tests.

View larger version (19K): [in a new window]   Figure 3. ELISA analysis demonstrating differences in TNF-, TNF-R1, and TNF-R2 levels in TMJ tissues 48 hrs after CFA treatment. Three different samples for both CFA-treated and control were homogenized, and total protein content was determined by Lowry assays. Each sample was run in triplicate, and the results are expressed as pmol/mg protein. Data are presented as means and standard deviations and were statistically analyzed by Student’s t tests. Significance (*) was determined at a level of p 0.05.