Release, biological potency, and biochemical integrity of recombinant human platelet-derived growth factor-BB (rhPDGF-BB) combined with Augment(TM) Bone Graft or GEM 21S beta-tricalcium phosphate (beta-TCP).
Over 10 million surgical procedures are performed annually in the United States to treat musculoskeletal injuries, and a significant portion of these involve orthopedic bone grafting. The goals of the study were to evaluate the in vitro and in vivo release kinetics, biological potency and biochemical integrity of rhPDGF-BB combined with large (1000-2000 microm) and small (250-1000 microm) beta-TCP particles. Recombinant human platelet-derived growth factor B homodimer (rhPDGF-BB) is a protein growth factor under development as a therapeutic for accelerating bone healing. Release of the protein was monitored in vitro by ELISA, and in vivo by measurement of radioactive rhPDGF-BB implanted in rat calvarial defects. Biological activity was measured using a cell-based bioassay, and biochemical integrity was determined by SDS-PAGE and high pressure size exclusion chromatography (HPSEC). Release of rhPDGF-BB occurred rapidly from beta-TCP both in vitro and in vivo. Almost 100% of the rhPDGF-BB was recovered from large and small beta-TCP after 90 min in vitro. Approximately 90% of the rhPDGF-BB was depleted from calvarial defect sites within 72 h of implantation. RhPDGF-BB retained 100% of its biological potency compared to reference standard rhPDGF-BB, manifested as a single band at ~30 kDa by SDS-PAGE and a single peak eluted after 13 min by HPSEC following release from beta-TCP. RhPDGF-BB is rapidly released from large and small beta-TCP particles and is biochemically unaltered following release.
Young
C S
CS
BioMimetic Therapeutics, Inc., 389 Nichol Mill Lane, Franklin, TN 37067, United States. cyoung@biomimetics.com
Ladd
P A
PA
Browning
C F
CF
Thompson
A
A
Bonomo
J
J
Shockley
K
K
Hart
C E
CE
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000077214:Becaplermin; D001672:Biocompatible Materials / Q000737:chemistry*; D002130:Calcium Phosphates / Q000008:administration & dosage / Q000737:chemistry*; D002850:Chromatography, Gel; D003692:Delayed-Action Preparations; D004337:Drug Carriers; D004343:Drug Implants; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D010982:Platelet-Derived Growth Factor / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000494:pharmacology*; D020574:Proto-Oncogene Proteins c-sis / Q000008:administration & dosage / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology; D012886:Skull / Q000502:physiology
D001672:Biocompatible Materials; D002130:Calcium Phosphates; D003692:Delayed-Action Preparations; D004337:Drug Carriers; D004343:Drug Implants; C000621424:GEM 21S; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; C485817:beta-tricalcium phosphate; D000077214:Becaplermin; D000469:Alkaline Phosphatase