Ex vivo 3D osteocyte network construction with primary murine bone cells

Osteocytes reside as three-dimensionally（3D） networked cells in the lacunocanalicular structure of bones and regulate bone and mineral homeostasis. Despite of their important regulatory roles, in vitro studies of osteocytes have been challenging because：（1） current cell lines do not sufficiently represent the phenotypic features of mature osteocytes and（2） primary cells rapidly differentiate to osteoblasts upon isolation. In this study, we used a 3D perfusion culture approach to：（1） construct the 3D cellular network of primary murine osteocytes by biomimetic assembly with microbeads and（2） reproduce ex vivo the phenotype of primary murine osteocytes, for the first time to our best knowledge. In order to enable 3D construction with a sufficient number of viable cells, we used a proliferated osteoblastic population of healthy cells outgrown from digested bone chips. The diameter of microbeads was controlled to：（1） distribute and entrap cells within the interstitial spaces between the microbeads and（2） maintain average cell-to-cell distance to be about 19 mm. The entrapped cells formed a 3D cellular network by extending and connecting their processes through openings between the microbeads. Also, with increasing culture time, the entrapped cells exhibited the characteristic gene expressions（SOST and FGF23） and nonproliferative behavior of mature osteocytes. In contrast, 2D-cultured cells continued their osteoblastic differentiation and proliferation. This 3D biomimetic approach is expected to provide a new means of：（1） studying flow-induced shear stress on the mechanotransduction function of primary osteocytes,（2） studying physiological functions of 3D-networked osteocytes with in vitro convenience,and（3） developing clinically relevant human bone disease models.

The gravity field and gravity data reduction across the continental area of Nigeria

This research presents the variation of the gravity field and associated gravity field components over the continental area of Nigeria to provide data for geoscience research,geodetic and engineering works,aerodynamic studies and deep crustal inferences.Accurate positions and elevations were observed at 58 of the 59 base stations of the Primary Gravity Network of Nigeria(PGNN),whose absolute gravity values had been accurately determined.The absolute gravity values were plotted against their respective positions to reveal the distribution pattern and strength of the gravity field within the study area.Theoretical gravity values at each base station were generated using the Somigliana's equation.The free-air gravity and free-air anomaly gravity values were generated with respect to the World Geodetic System 1984(WGS84)ellipsoid using GPS-derived elevation data.Then,the perturbing potential,free-air gravity with respect to the geoid,and the indirect effects were evaluated.The average of the indirect effects was used to adjust the WGS84 gravity formula to produce a gravity formula that better approximates the geoid across the continental area of Nigeria,compatible with the heights measured relative to the geoid,which can serve as a reference for establishing a vertical height control.The Bouguer gravity and Bouguer gravity anomalies across Nigeria revealed a“trans-southern gravity high strip”interpreted to be associated with mantle upwelling.Two new major mega-lineaments related to mantle upwelling were mapped.A batholith province trending NWeSE was delineated,occurring from north central Nigeria to the north western region and containing closures of“Bouguer gravity lows”interpreted as batholiths.A separate closure of“Bouguer gravity low”was detected at Azare,north eastern Nigeria,which may be due to the presence of intrusive granitic body.It is recommended that the mantle structure beneath“the trans-southern gravity high strip”,“delineated batholith province”and“isolated gravity closures”around the northeast of Nigeria should be studied from seismic shear wave splitting analysis for better understanding of the deep lithospheric structures and moho relief.