Delivery of transforming growth factor-beta2-perturbing antibody in a collagen vehicle inhibits cranial suture fusion in calvarial organ culture.
OBJECTIVE
To determine whether antibody perturbation of Tgf-beta, delivered in a collagen gel, could inhibit cranial suture fusion.


METHODS
Attachment and proliferation of osteoblasts cultured on a collagen gel with or without anti-Tgf-beta2 antibody were determined by AlamarBlue dye assay and cell morphology by toluidine-blue staining. In rat calvarial organ culture, collagen gel with and without anti-Tgf-beta2 antibody was injected subperiosteally over the posterior frontal suture of postnatal day 15 rat calvariae. A quantitative analysis of suture fusion was used to measure suture bridging in histological serial sections at various time points.


RESULTS
Attachment and proliferation for cells cultured on collagen gel with anti-Tgf-beta2 antibody were similar to collagen gel controls. Although proliferation was lower than on tissue culture plastic, cells treated with anti-Tgf-beta2 antibody maintained an osteoblastic morphology. After 7, 10, and 15 days in organ culture, anti-Tgf-beta2 antibody treatment caused a reduction in the percent bridging of posterior frontal sutures, compared with controls. Sutures exposed to anti-Tgf-beta2 antibody and fibroblast growth factor-2 concurrently did not show an inhibition of bony bridging.


CONCLUSIONS
These results support previous reports suggesting a role for Tgf-beta2 in cranial suture fusion. In cell culture the collagen gel, both with and without anti-Tgf-beta2 antibody, promoted similar osteoblast attachment, proliferation, and osteoblastic morphology. In organ culture anti-Tgf-beta2 antibody was delivered in a bioactive state via a collagen gel to inhibit cranial suture fusion. Also, the results suggest that the inductive effect of fibroblast growth factor-2 is not dependent on Tgf-beta2 activity. Together, these results provide further support for the role of Tgf-beta2 in cranial suture fusion.
Department of Pediatric Dentistry, College of Dentistry, Ohio State University, Postle Hall, Room 4126, 305 West 12th Avenue, PO Box 182357, Columbus, OH 43218-2357, USA. moursi.1@osu.edu
Amr M
AM
Moursi
Phillip L
PL
Winnard
Doug
D
Fryer
Mark P
MP
Mooney
D000704:Analysis of Variance; D000818:Animals; D000906:Antibodies / Q000008:administration & dosage; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen; D003393:Cranial Sutures / Q000187:drug effects*; D016222:Fibroblast Growth Factor 2 / Q000502:physiology; D005782:Gels; D009924:Organ Culture Techniques; D010006:Osteoblasts / Q000187:drug effects; D014677:Pharmaceutical Vehicles; D051381:Rats; D017207:Rats, Sprague-Dawley; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000502:physiology*; D053781:Transforming Growth Factor beta2
D000906:Antibodies; D005782:Gels; D014677:Pharmaceutical Vehicles; D016212:Transforming Growth Factor beta; D053781:Transforming Growth Factor beta2; D016222:Fibroblast Growth Factor 2; D003094:Collagen