Ionophore and Biometal Modulation of P-glycoprotein Expression and Function in Human Brain Microvascular Endothelial Cells

Purpose

Biometals such as zinc and copper have been shown to affect tight junction expression and subsequently blood-brain barrier (BBB) integrity. Whether these biometals also influence the expression and function of BBB transporters such as P-glycoprotein (P-gp) however is currently unknown.

Methods

Using the immortalised human cerebral microvascular endothelial (hCMEC/D3) cell line, an in-cell western assay (alongside western blotting) assessed relative P-gp expression after treatment with the metal ionophore clioquinol and biometals zinc and copper. The fluorescent P-gp substrate rhodamine-123 was employed to observe functional modulation, and inductively coupled plasma mass spectrometry (ICP-MS) provided information on biometal trafficking.

Results

A 24-h treatment with clioquinol, zinc and copper (0.5, 0.5 and 0.1 μM) induced a significant upregulation of P-gp (1.7-fold) assessed by in-cell western and this was confirmed with western blotting (1.8-fold increase). This same treatment resulted in a 23% decrease in rhodamine-123 accumulation over a 1 h incubation. ICP-MS demonstrated that while t8his combination treatment had no effect on intracellular zinc concentrations, the treatment significantly enhanced bioavailable copper (4.6-fold).

Conclusions

Enhanced delivery of copper to human brain microvascular endothelial cells is associated with enhanced expression and function of the important efflux pump P-gp, which may provide therapeutic opportunities for P-gp modulation.

     

A ganglion of the temporomandibular joint: a case report

Ganglion cysts of the TMJ are rare and accurate preoperative diagnosis is not usually made, although theoretically this might be possible with arthrography. Treatment of the condition is surgical but, if a positive diagnosis can be made, a period of conservative management could be attempted if the cyst is not excessively painful or surgery is contraindicated for any reason.     

Pulp changes after anterior mandibular subapical osteotomy in a primate model.

Subapical osteotomy of the anterior mandible was carried out on 10 adult Macaca Irus monkeys with and without open bite. Teeth were extracted at 8 weeks, 12 weeks, 24 weeks and 1 year after surgery and the pulps examined histologically. No normal pulps were found. Progressive fibrosis and calcification was noted in all teeth. Patent pulp blood vessels were found up to 24 weeks after surgery. Pulp damage to teeth distal to the osteotomy site was found in 50% of cases. Control teeth extracted from the opposing jaw at the same intervals were all found to have normal pulps.