Biocompatibility of a novel biological piezoelectric ceramic to the rat periosteum derived osteoblast / 华西口腔医学杂志

<p><b>OBJECTIVE</b>A novel biological piezoelectric ceramic was made by beta-tricalcium phosphate (beta-TCP) and lithium sodium potassium niobate (LNK) piezoelectric ceramics. To study its biocompatibility to osteoblast isolated from the cranium of 1-day-old Sprague-Dawley mice.</p><p><b>METHODS</b>The biological piezoelectric ceramic TCPLNK1/10, TCPLNK5/5 respectively mixed by beta-TCP and LNK piezoelectric ceramic at the ratio of 1/10 and 5/5. Then osteoblasts were used and seeded respectively on the negative and positive surfaces of TCPLNK1/10 and TCPLNK5/5. Growth and proliferation of the osteoblasts on TCPLNK1/10 and TCPLNK5/5 surfaces were evaluated in vitro by means of scanning electron microscopy (SEM) examination, fluorescence dyeing of osteoblast skeleton protein and MTT assay.</p><p><b>RESULTS</b>Cell morphology of osteoblast on positive and negative surfaces of TCPLNK1/10 and TCPLNK5/5 was normal, and both adhesion and growth characteristics showed better than control group. The growing osteoblasts on the TCPLNK1/10 negative surface were significantly higher than others. The negative surface of TCPLNK1/10 possessed better osteogenesis potential than others in vitro.</p><p><b>CONCLUSION</b>The surface of TCPLNK may permit the imitation piezoelectric effect of natural bone for bone regeneration.</p>

Recombinant expression and secretion of mpd gene using the promoters and signal peptide-encoding sequences of ytkA and ywoF gene from Bacillus subtilis / 生物工程学报

A shuttle promoter-probe vector pNW33N-mpd was constructed with the E. coli-B. subtilis shuttle vector pNW33N and the mature mpd gene without it' s signal peptide-encoding sequence. The promoter fragments of B. subtilis ytkA and ywoF gene were cloned from plasmid pMPDP3 and pMPDP29 then generated the shuttle expression vector pNYTM and pNYWM. Expression vectors pNYTM and pNYWM were transformed into B. subtilis 1A751 to construct the expression strain 1A751 (pNYTM) and 1A751 (pNYTM), in these strains, under the control of the promoters and signal peptides of ytkA and ywoF gene, mpd gene was expressed and secreted with its biological activity; the result showed that the promoter of ytkA gene is much stronger than that of ywoF gene. Then a new shuttle expression-secretion vector pYNMK was constructed using the ytkA gene promoter and the signal peptide-encoding sequence of B. subtilis nprB gene, the expression of mpd gene achieved a higher level using the B. subtilis WB800 as the host, the methyl parathion hydrolase activity accumulated to a maximum level of 10.40 u/mL after 84 h of cultivation at the late stationary phase, which was 10.8-fold higher than the expression level of the original Plesiomonas strain M6, about 91.4% of the recombinant expression production was secreted into the culture medium.