Porphyromonas gingivalis Lipopolysaccharide-Induced B Cell Differentiation by Toll-like Receptors 2 and 4.

BACKGROUND: Porphyromonas gingivalis (P. gingivalis) is a pathogenic bacterium widely present in subgingival plaques of patients with periodontitis. It induces periodontitis with bone loss as its main feature by changing the number and composition of symbiotic microorganisms, as well as inducing the natural immune response of the host. However, the mechanism of the latter remains unclear. OBJECTIVE: This study aims to investigate the effect of P. gingivalis lipopolysaccharide (LPS) on regulatory B cells (Breg) in the occurrence and development of periodontitis. METHODS: We detected the mRNA levels of IL-10 in B cells under the stimulation of P. gingivalis LPS and/or E. coli LPS, distinguished IL-10-producing cells from different B cell subgroups using flow cytometry. Through toll-like receptor (TLR) knockout mice, the role of TLR2 and TLR4 in this process was also evaluated. RESULTS: Results showed that P. gingivalis stimulated B cells to produce IL-10 via TLR2/4. CD5+B1 subset is the main source of IL-10+Breg cell. Under P. gingivalis LPS stimulation, CD5+IgM+CD93-IL-10+B cell subset increased significantly, which was regulated through TLR2/ 4. CONCLUSION: The results of this study provides new insights into the immunopathogenic mechanism of P. gingivalis, preliminarily discussed the effect of P. gingivalis on the production of Breg, and present a theoretical foundation for subsequent investigations on the occurrence and development of periodontitis.

A modified collagen scaffold facilitates endogenous neurogenesis for acute spinal cord injury repair.

Due to irreversible neuronal loss and glial scar deposition, spinal cord injury (SCI) ultimately results in permanent neurological dysfunction. Neuronal regeneration of neural stem cells (NSCs) residing in the spinal cord could be an ideal strategy for replenishing the lost neurons and restore function. However, many myelin-associated inhibitors in the SCI microenvironment limit the ability of spinal cord NSCs to regenerate into neurons. Here, a linearly ordered collagen scaffold was used to prevent scar deposition, guide nerve regeneration and carry drugs to neutralize the inhibitory molecules. A collagen-binding EGFR antibody Fab fragment, CBD-Fab, was constructed to neutralize the myelin inhibitory molecules, which was demonstrated to promote neuronal differentiation and neurite outgrowth under myelin in vitro. This fragment could also specifically bind to the collagen and undergo sustained release from collagen scaffold. Then, the scaffolds modified with CBD-Fab were transplanted into an acute rat SCI model. The robust neurogenesis of endogenous injury-activated NSCs was observed, and these NSCs could not only differentiate into neurons but further mature into functional neurons to reconnect the injured gap. The results indicated that the modified collagen scaffold could be an ideal candidate for spinal cord regeneration after acute SCI. STATEMENTS OF SIGNIFICANCE: A linearly ordered collagen scaffold was specifically modified with collagen-binding EGFR antibody, allowed for sustained release of this EGFR neutralizing factor, to block the myelin associated inhibitory molecules and guide spinal cord regeneration along its linear fibers. Dorsal root ganglion neurons and neural stem cells induced by CBD-Fab exhibited enhanced neurite outgrowth and neuronal differentiation rate under myelin in vitro. Transplantation of the modified collagen scaffold with moderate EGFR neutralizing proteins showed greatest advantage on endogenous neurogenesis of injury-activated neural stem cells for acute spinal cord injury repair.

3D bioprinting of BMSC-laden methacrylamide gelatin scaffolds with CBD-BMP2-collagen microfibers.

Three-dimensional (3D) bioprinting combines biomaterials, cells and functional components into complex living tissues. Herein, we assembled function-control modules into cell-laden scaffolds using 3D bioprinting. A customized 3D printer was able to tune the microstructure of printed bone mesenchymal stem cell (BMSC)-laden methacrylamide gelatin scaffolds at the micrometer scale. For example, the pore size was adjusted to 282 ± 32 µm and 363 ± 60 µm. To match the requirements of the printing nozzle, collagen microfibers with a length of 22 ± 13 µm were prepared with a high-speed crusher. Collagen microfibers bound bone morphogenetic protein 2 (BMP2) with a collagen binding domain (CBD) as differentiation-control module, from which BMP2 was able to be controllably released. The differentiation behaviors of BMSCs in the printed scaffolds were compared in three microenvironments: samples without CBD-BMP2-collagen microfibers in the growth medium, samples without microfibers in the osteogenic medium and samples with microfibers in the growth medium. The results indicated that BMSCs showed high cell viability (>90%) during printing; CBD-BMP2-collagen microfibers induced BMSC differentiation into osteocytes within 14 days more efficiently than the osteogenic medium. Our studies suggest that these function-control modules are attractive biomaterials and have potential applications in 3D bioprinting.

[Distribution, seasonal variation and influence factors of dissolved inorganic arsenic in the Sanggou Bay].

The biogeochemical behavior of arsenic in the aquatic environment has already captured the attentions of scientists due to its complex forms and toxicity. Four cruises were carried out in April, August, October 2011 and January 2012 in the Sanggou Bay. The concentrations of total dissolved inorganic arsenic (TDIAs, TDIAs = [ As(5+] + [As(3+)]) and arsenite (As(3+)) were measured by Hydride Generation-Atomic Fluorescence Spectrometry (HG-AFS). The concentrations of TDIAs ranged from 3.4-12.4 nmol x L(-1) in April, 8.9-16.9 nmol x L(-1) in August, 14.7-21.3 nmol x L(-1) in October and 13.8-21.9 nmol x L(-1) in January. The concentrations of arsenite ranged from 0.3-2.1 nmol x L(-1), 0.4-3.8 nmol x L(-1), 1.8-4.0 nmol x L(-1) and 0.3-2.9 nmol x L(-1) during four cruises, respectively. The concentrations of TDIAs in spring and summer were lower than those in autumn and winter, and high values of TDIAs appeared in the bay-mouth and the coastal estuary. The concentrations of arsenite in spring and winter were lower than those in summer and autumn. The maximum As(3+)/TDIAs ratios appeared in summer. The mean value of TDIAs in the Sanggou Bay was (13.9 +/- 4.7) nmol x L(-1), which was lower than the national primary drinking in water Standards from USEPA and met the first grade water quality based on the environmental quality standards for surface water of China. It indicates that there is no obvious anthropogenic pollution. The concentrations of TDIAs in the Sanggou Bay were lower than those in the Ailian Bay and the Lidao Bay in spring and summer due to the different hydrological environments and terrestrial inputs. Riverine input, incursion of Yellow Sea and biological activities were the three main factors impacting the distribution of TDIAs in the Sanggou Bay, and the influence of aquaculture activities was particularly significant. The enrichment of arsenic by aquaculture may lead to potential ecological crisis and food safety problems, and need to be paid more attentions to ensure the sustainable development of aquaculture in the Sanggou Bay.

[Distribution and air-sea fluxes of methane in the Yellow Sea and the East China Sea in the spring].

A survey was carried out in the Yellow Sea and the East China Sea from March 17 to April 06 of 2011. Dissolved CH4 in various depths were measured and sea-to-air fluxes were estimated. Methane concentrations in surface and bottom waters ranged between 2.39-29.67 nmol x L(-1) and 2.63-30.63 nmol x L(-1), respectively. Methane concentrations in bottom waters were slightly higher than those in surface waters, suggesting the existence of methane source in bottom waters or sediments. The horizontal distribution of dissolved CH4 showed a decrease from the river mouth to the open sea, and was influenced by the freshwater discharge and the Kuroshio intrusion. Surface methane saturations ranged from 93%-1 038%. Sea to air CH4 fluxes were (2.85 +/- 5.11) micromol x (m2 x d)(-1) (5.18 +/- 9.99) micromol x (m2 x d)(-1) respectively, calculated using the Liss and Merlivat (LM86), the Wanninkhof (W92) relationships and in situ wind speeds, and estimated emission rates of methane from the East China Sea and the Yellow Sea range from 7.05 x 10(-2) - 12.0 x 10(-2) Tg x a(-1) and 1.17 x 10(-2) - 2.20 x 10(-2) Tg x a(-1), respectively. The Yellow Sea and East China Sea are the net sources of atmospheric methane in the spring.