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Morphological osteoblasts were observed.
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For morphological observation, osteoblasts were seeded on the control and modified surfaces at a density of 4 × 10 cells/cm and examined with scanning electron microscopy (SEM).
Figure 4 shows the morphologies of osteoblasts on different regions.
As FoxO is a critical protein in canonical hedgehog signalling, we decided to explore the possible involvement of hedgehog signalling during osteoblast morphological changes.
Functional confirmation of the relevance of these results for osteoblast morphological transitions came from experiments in which Shh hedgehog signalling was inhibited using the well-established pathway inhibitor cyclopamine (Cyc).
The results obtained were used to construct interactomes, and these revealed an important role for FoxO in mediating morphological changes of osteoblast, which was validated by Western blot technology when FoxO was significantly up-expressed in response to Matrigel™.
This indicates that Ca2+ is required for the redistribution of focal adhesions and the actin cytoskeleton that occurs as these osteoblasts undergo morphological change during migration.
Moreover, the osteoblasts undergo morphological changes, becoming large and cuboidal cells [ 26, 29– 31].
TRAP staining of osteoclasts on histological sections (3 μm) and identification of osteoblasts using morphological criteria were done as previously described [11].
Our transmission electron microscopy study of cells located near HA and Mg-HAp microcalcifications revealed the presence of cells with morphological characteristics typical of osteoblasts.
The reciprocal and adaptive interactions between cells and substrates governing morphological transitions in the osteoblast compartment remain largely obscure.
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