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To determine the IL-6 levels in the synovial fluids of healthy, defect and OA donors and in the conditioned media of healthy, defect and OA chondrocytes during regeneration, a multiplex ELISA was performed as previously described [ 50, 51].
Isolated chondrocytes from healthy, defect and OA cartilage were used either directly after isolation or after expansion to passage 2 (see Table 1 for more details).
A total of 12 cytokines were measured of which IL-6 was most differentially regulated by healthy, defect and OA donors and hence chosen for further investigation.
To determine the cytokine levels in the synovial fluids and tissue extracts of healthy, defect and OA donors and in the conditioned media (day 7) of healthy, defect and OA chondrocytes during in vitro tissue formation, a multiplex bead-assay (Luminex, Luminex Corporation, Austin TX, USA) was performed as described previously [ 13, 14].
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Although popularity usually attracts commercial breeders who mass produce poor-quality dogs, I would hope that buyers would deal only with responsible breeders trying to produce healthy, defect-free dogs.
As no difference was found in IL-6 production between healthy and defect chondrocytes, only defect and osteoarthritic chondrocytes were studied.
Total GAG production per DNA was not different between healthy, cartilage defect and OA nonexpanded chondrocytes.
In healthy and defect chondrocytes endogenous IL-6 production was much lower than in OA chondrocytes.
There was no significant difference in IL-6 production between healthy and defect chondrocytes.
There were no differences in DNA content between healthy, cartilage defect and OA tissue (P = 0.1) (Table 3).
IL-6 production of osteoarthritic chondrocytes during cartilage regeneration was higher than that of healthy and defect chondrocytes (P < 0.001).
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