D-alanine suppressed osteoclastogenesis derived from bone marrow macrophages and downregulated ERK/p38 signalling pathways.

OBJECTIVES: D-alanine is a residue of the backbone structure of Type Ⅰ Lipoteichoic acid (LTA), which is a virulence factor in inflammation caused by gram-positive bacteria. However, the role of D-alanine in infectious bone destruction has not been investigated. We aimed to explore the role of D-alanine in the proliferation, apoptosis, and differentiation of osteoclasts. DESIGN: Mouse bone marrow-derived macrophages (BMMs) were isolated as osteoclast precursors and stimulated with D-alanine. Cell proliferation and apoptosis were detected using CCK-8 and flow cytometry, respectively. The formation of osteoclasts morphologically observed by tartrate-resistant acid phosphatase staining (TRAP) and immunofluorescence staining. The expressions of osteoclastogenic genes were measured by real-time RT-PCR. The protein expressions of osteoclastogenic markers, p38, and ERK1/2 MAPK signalling were measured by western blot. The expression level of soluble Sema4D was detected via enzyme-linked immunosorbent assay (ELISA). RESULTS: The cell proliferation of BMMs was significantly inhibited by D-alanine in a dose-dependent manner. Apoptosis of BMMs was markedly activated with the stimulation of D-alanine. The differentiation of BMMs into osteoclasts was significantly inhibited by D-alanine, and the gene and protein expressions of NFATc1, c-Fos, and Blimp decreased. Western blot showed that D-alanine inhibited the phosphorylated p38 and ERK1/2 signalling pathways of BMMs. Moreover, the expression level of soluble Sema4D significantly decreased in the supernatant of BMMs due to the D-alanine intervention. CONCLUSION: D-alanine plays a pivotal role in the inhibition of RANKL-induced osteoclastogenesis and might become a potential therapeutic drug for bone-resorptive diseases.

Effect of Enterococcus faecalis on osteoclastogenesis under cobalt-mimicked hypoxia in vitro.

OBJECTIVE: The bone destruction in persistent apical periodontitis associated with infection and a periapical hypoxic microenvironment is not well known. Thus, we aimed to investigate the effects of Enterococcus faecalis on osteoclastogenesis under cobalt-mimicked hypoxia. MATERIALS AND METHODS: Mouse bone marrow-derived macrophages (BMMs) were isolated as osteoclast precursors and stimulated by heat-killed E. faecalis in an environment of cobalt-mimicked hypoxia environment. The cell proliferation and apoptosis were detected using CCK-8 and flow cytometry, respectively. Osteoclast differentiation was determined via tartrate-resistant acid phosphatase staining (TRAP) and immunofluorescence staining. The osteoclastogenic protein and gene expressions were measured by western blotting and real-time PCR. RESULTS: Under cobalt-mimicked hypoxia, E. faecalis markedly inhibited the proliferation of the BMMs and significantly promoted the apoptosis of the BMMs. The differentiation of the BMMs into osteoclasts was enhanced in the presence of the E. faecalis under hypoxia, and the expression of Blimp, c-Fos, and NFATc1 was up-regulated, while the expression of RBP-J was inhibited. CONCLUSIONS: E. faecalis markedly promotes osteoclast differentiation under cobalt-mimicked hypoxia in vitro.

A Photonic crystal fiber with large effective refractive index separation and low dispersion.

A photonic crystal fiber (PCF) structure with a ring-core and 5 well-ordered semiellipse air-holes has been creatively proposed. Through a comparison between the structures with a high refractive index (RI) ring-core and the structure without, it conclude that a PCF with a high RI ring-core can work better. Schott SF57 was elected as the substrate material of ring-core. This paper compares the effects of long-axis and short-axis changes on the PCF and selects the optimal solution. Especially TE0,1 mode's dispersion is maintained between 0 and 3 ps / (nm · km) ranging from 1.45 µm to 1.65 µm. This property can be used to generate a supercontinuum with 200 µm long zero dispersion wavelength (ZDM). In addition, Δneff reaches up to 10-3, which enables the near -degeneracy of the eigenmodes to be almost neglected. The proposed PCF structure will have great application value in the field of optical communications.

Simultaneous measurement of optical rotation dispersion and absorption spectra for chiral substances.

We present a new method based on optical null methods for simultaneously measuring the optical rotatory dispersion (ORD) and absorption spectra of chiral substances. The optical rotation angle at a specific wavelength can be obtained from the optical nulls of the Malus curves with and without the sample. We use the optical nulls of the two curves as benchmark points and the readings to the right of the benchmark points by a certain angular offset to eliminate the influence of the analyzer on the light intensity and obtain the absorbance of the chiral substance at a specific wavelength. The 4096 pixels of the line scan CCD can measure multiple wavelengths simultaneously so that continuous ORD and absorption spectra can be obtained. The experimental results show that the standard deviation of the specific optical rotation is 0.11 deg mL g-1 dm-1 , the standard deviation of the maximum absorption wavelength is 0.45 nm, and that absorbance of the sample varies linearly with the concentration. This method is helpful for simplifying the experiment and has a profound influence on the analysis of the contents and molecular configurations of chiral substances in the future.