Osteogenic Evaluation of GTG Composite with Fetal Rat Calvar ial Culture Model
Date Issued
1998-07-31
Date
1998-07-31
Author(s)
林輝
DOI
872314B002310M08
Abstract
The cytotoxicity of the synthetic bone substitute composed of tricalcium phosphate
and glutaraldehyde cross-linked gelatin (GTG) has been evaluated by osteoblasts cell
culture. In the previous study, the GTG composites have been soaked in distilled
water for 1, 2, 4, 7, 14, 28, and 42 days and then the solutions (or extracts) were
cocultured with osteoblasts to evaluate the cytotoxicity of GTG composites by alive
cell counting.
In the study, the extracts were cocultured with the osteoblasts; thereafter the
concentration of TGF-b1 and PGE2 in the medium was analyzed to strictly reflect the
biological effects of GTG composites on the growth of osteoblasts. In order to
investigate the osteoconductive potential of the GTG composites on new bone
formation in a relative short-term, a model of neonatal rat calvarial organ culture
was designed prior to animal experiment. Three experimental materials of 4%, 8%,
and 12% GTG composites were evaluated by fetal rat calvarial organ culture for
their ability for bone regeneration. Deproteinized bovine and porcine cancellous bone
matrixes were used as the controlled materials. All the organ culture units were
maintained in cultured medium for 5 weeks. Following the culture period, the
morphology of tissue was observed under optical microscope and the quantitative
evaluation of the new generation bone was determined by using a semiautomatic
histomorphometeric method.
Except in the initial 4 days, the concentration of TGF-b1 of 4% and 8% GTG
composites was higher than that of the blank group for all the other experimental time
period. The PGE2 concentration for 4% and 8% GTG composites was lower than that
of the blank group. It revealed that the 4% and 8% GTG composites would not lead to
inflammation and would promote the osteoblast growth. The morphology and activity
of the osteoblasts were not transformed or changed by the two GTG composites. For
the 12% GTG composite, the performance of in vitro condition inferior to the blank
group and the other two GTG composites. Though the concentration of TGF-b1 and
PGE2 was gradually back to normal after 14 days, the morphology of the
osteoblasts was abnormal with features such as contracted cytoplast structure. The
osteoblast perhaps be damaged in the initial stage. We suggested that the 4% and 8%
GTG composites should be soaked in distilled water at least for 4 days before medical
applications. The 12% GTG composite or the composites with concentration of
glutaraldehyde solution higher than 12% was not recommended as the medical
prostheses in any condition. The fetal rat calvaria culture also showed the same
results with the analysis of TGF-b1 and PGE2. From the study, we could use to
predict the results of animal experiment in the future.
Subjects
bioabsorbed bone graft
glutaraldehyde
tricalcium phosphate
gelatin
osteoblast
rat calvarial tissue culture
Publisher
臺北市：國立臺灣大學醫學工程學研究所
Type
report
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