A major limitation in the bio-medical sector is the availability of materials suitable for bone tissue engineering using stem cells and methodology converting the stochastic biological events towards definitive as well as efficient bio-mineralization. We show that osteoblasts and Bone Marrow-derived Mesenchymal Stem Cell Pools (BM-MSCP) express TRPM8, a Ca2+-ion channel critical for bone-mineralization. TRPM8 inhibition triggers up-regulation of key osteogenesis factors; and increases mineralization by osteoblasts. We utilized CMT:HEMA, a carbohydrate polymer-based hydrogel that has nanofiber-like structure suitable for optimum delivery of TRPM8-specific activators or inhibitors. This hydrogel is ideal for proper adhesion, growth, and differentiation of osteoblast cell lines, primary osteoblasts, and BM-MSCP. CMT:HEMA coated with AMTB (TRPM8 inhibitor) induces differentiation of BM-MSCP into osteoblasts and subsequent mineralization in a dose-dependent manner. Prolonged and optimum inhibition of TRPM8 by AMTB released from the gels results in upregulation of osteogenic markers. We propose that AMTB-coated CMT:HEMA can be used as a tunable surface for bone tissue engineering. These findings may have broad implications in different bio-medical sectors.

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An interlaboratory study was conducted to evaluate a method for the determination of campesterol, stigmasterol, and beta-sitosterol in saw palmetto raw materials and dietary supplements at levels >1.00 mg/100 g based on a 23 g sample. Test samples were saponified at high temperature with ethanolic KOH solution. The unsaponifiable fraction containing phytosterols (campesterol, stigmasterol, and beta-sitosterol) was extracted with toluene. Phytosterols were derivatized to trimethylsilyl ethers and then quantified by gas chromatography with hydrogen flame ionization detection. Twelve blind duplicates, one of which was fortified, were successfully analyzed by 10 collaborators. Recoveries were obtained for the sample that was fortified. The results were 99.8, 111, and 111% for campesterol, stigmasterol, and beta-sitosterol, respectively. For repeatability, the relative standard deviation (RSDr) ranged from 3.93 to 17.3% for campesterol, 3.56 to 22.7% for stigmasterol, and 3.70 to 43.9% for beta-sitosterol. For reproducibility, the RSDR ranged from 7.97 to 22.6%, 0 to 26.7%, and 5.27 to 43.9% for campesterol, stigmasterol, and beta-sitosterol, respectively. Overall, the Study Director approved 5 materials with acceptable HorRat values for campesterol, stigmasterol, and beta-sitosterol ranging from 1.02 to 2.16.