Characterization of a fibrin glue-GDNF slow-release preparation.
One novel method to deliver trophic factor locally in the CNS is to mix it into fibrin glue. In the present studies, [125I]-labeled GDNF-containing fibrin glue balls were used to determine binding and spread of the trophic factor. First, the binding of different concentrations of [125I]-labeled GDNF in fibrin glue was determined in vitro. Within the six concentrations used (from 200 nM to 0.004 nM, 0 M as control), there was a strong linear correlation between the [125I]-GDNF concentration and the recovered radioactivity (r = 0.992). The mean bound radioactivity in 16 samples with 4 nM [125I]-GDNF was 71262 +/- 2710 CPM, and accounted for 89.8% of the mean initial count of free [125I]-GDNF (79369 +/- 3499 CPM). Second, [125I]-GDNF-containing glue balls were implanted into the anterior chamber of adult rats. The implanted fibrin glue balls decreased in size with time, but could still be identified on the irises 2 wk after implantation. Radioactivity was concentrated at the implantation sites in the early stages with a distribution in the surrounding iris tissue, which became separated into focal radioactive spots at the third week. Counts of radioactivity were significantly higher in the [125I]-GDNF glue ball-implanted irises than controls until 14 days after implantation. A study of the [125I] decay over time using least-squares linear regression demonstrated first-order kinetics (r = -0.98, p <0.02) with k = 0.0091 and T 1/2 = 76 h. Finally, [125I]-GDNF-containing glue balls were implanted in the spinal cord of adult rats. Radioactivity was concentrated at the implantation sites in the early stages and was later distributed more widely in the surrounding thoracic cord. The [125I]-GDNF-containing glue degraded over time and became a porous meshwork with decreasing radioactivity at the later time points. Radioactivity in the spinal cords subjected to implantation of [125I]-GDNF-containing glue balls was higher than in controls for 14 days. Study of the [125I] decay by time with least-squares linear regression demonstrated first-order kinetics (r = -0.97, p = 0.001) with T 1/2 = 75.6 h. We conclude that the trophic factor GDNF becomes bound in the fibrin glue matrix from which it is gradually released. Our results suggest that fibrin glue is an effective substrate for keeping a trophic factor localized in situ for a finite period, protected from the circulation, surrounding aqueous humor or CSF.
Cheng
H
H
Department of Neuroscience, Karolinska Institute, Stockholm, Sweden.
Fraidakis
M
M
Blombäck
B
B
Lapchak
P
P
Hoffer
B
B
Olson
L
L
D000818:Animals; D000867:Anterior Chamber / Q000187:drug effects / Q000378:metabolism; D001345:Autoradiography; D003692:Delayed-Action Preparations; D005260:Female; D015718:Fibrin Tissue Adhesive / Q000008:administration & dosage*; D051100:Glial Cell Line-Derived Neurotrophic Factor; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D009414:Nerve Growth Factors / Q000008:administration & dosage* / Q000493:pharmacokinetics; D009419:Nerve Tissue Proteins / Q000008:administration & dosage* / Q000493:pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism; D014014:Tissue Adhesives / Q000494:pharmacology*
D003692:Delayed-Action Preparations; D015718:Fibrin Tissue Adhesive; C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D007457:Iodine Radioisotopes; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D014014:Tissue Adhesives