loaded samples were placed into the kappa mu Opioid Receptor reactor and a bioreactor-built mechanical stimulus compression, as reported previously in our laboratory. In particular, dynamically compressed samples by a strain of 10% of sine Dale anf with Nglichen load of 5% for a period of 6 h stimulated. The control groups were obtained in the incubator for the same duration as the tablet samples. 2.4. RNA isolation and cDNA synthesis immediately after the compression stroke for 6 h, the bioreactor was disassembled and all samples were run in TRIzol reagent in a position to the method of RNA isolation. RNA was isolated according to the manufacturer’s protocol and RNA pellets were resuspended in st RNase, DNase-free water gel. Then, using a spectrophotometer case nano-RNA concentration determined for each sample. A kit with high-capacity t reverse transcription was used for cDNA synthesis, and followed the manufacturer protocol. Briefly, each reaction for a lg RNA in a total volume of 10 ll was mixed with 10 ll reaction buffer ma 10 RT buffer, 25 dNTP mix, 10 Feeder Llige primer RT, reverse transcriptase MultiScribe exist, RNase inhibitor and nuclease-free water. All reaction tubes were then placed in a thermal cycler and the following temperature cycle was required as described by the kit. Zun Were kept Highest all samples at 25 ° C for 10 min and then heated at 37 ° C for 120 min. Close Lich, the reaction by increasing Increase of the sample to 85 ° C was stopped for 5 min, then stored at 4 ° C 2.5. Standard polymerase chain reaction to analyze gene expression, a standard procedure for the Warmth No polymerase reaction was monitored. Briefly, cDNA mixture diluted to 10 ng / ll and 20 ng of total new sterile Hrchen R And were transferred to the PCR cocktail. Together for each reaction, a cocktail of 10 high-fidelity PCR buffer, 10 mM dNTP mix, 50 mM MgSO 4, specific primer pairs of genes, Platinum Taq DNA Polymerase High Fidelity and nuclease-free water. Subsequently, the reaction tubes placed in a thermal cycler and the reaction was carried out as follows: denaturation at 94 ° C for 30 s, annealing step, with the exception of collagen type I and Angiotensin osteocalcin at 55 ° C for 45 seconds and extension at 72 ° C for 1 min. In this way gave it a further min hold at 72 ° C for 10, then the samples were stored at 4 ° C. The results were the PCR reactions were performed by gel electrophoresis in a 2% agarose gels in Tris / borate / EDTA with ethidium bromide analyzes produced added. PCR reactions were then mixed with a dye bromophenol blue loading DNA samples before loading into the gels. After loading a voltage was applied from 120 V to gels about 45 minutes, are then placed on a UV light source. The integrated density values for each amplicon was measured and analyzed the system and the simple ChemiImager 鈩 Alpha Imaging software. The expressions of genes for all samples were housekeeping gene glyceraldehyde phosphate dehydrogenase, and three samples of controlled group converted On within each gene of interest. 2.6. Histochemical F Staining and fluorescence additives Tzlich samples within each experimental group were also of c Tea for Ritonavir histochemical F Staining. at the end of the compression of 6 h, the gels staini.