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Here, we demonstrate the long-term biocompatibility of 4.5 μm RGD-coated particles with osteoblasts up to 21 days in culture, and detail a time course of responses elicited from osteoblasts following mechanical stimulation with integrin attached magnetic particles (<2 h post attachment) and internalised particles (>48 h post attachment).
Validation of this scoring method was carried out by monitoring cells post attachment.
Starting from 24 hours post attachment, permanent byssal threads become dominant [ 28].
Based on what has been observed in previous studies, within 12 hour post attachment, the temporary byssal threads are rapidly produced.
Fed male ticks were collected from several feeding experiments, while female ticks were manually detached at 24 and 96 h post attachment and processed for RNA extraction as described below.
We are interested in 24-48 h post attachment tick saliva proteins because this tick feeding stage precedes some of the most important facets of tick parasitism, blood meal feeding, transmission, and acquisition of tick borne disease agents.
Ticks of different sizes, but always ≤ 4 mm in body length (before the rapid engorgement phase of feeding; approximately three to four days post attachment) were used for salivary gland dissection to avoid degeneration of salivary gland proteins [ 36, 37].
Just a faint signal was obtained in RNA from Cuscuta shoots lacking haustorial structures and a strong one was obtained in RNA from shoot material with haustoria, which was harvested at three days post attachment.
To measure a shift in the PWV due to cell attachment, the sensor surface is scanned twice: before (baseline scan) and after cells have attached to the surface (post-attachment scan).
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