Objective To explore the regulator factor of osteogenes is induced by the fibroblast in vitro so as to provide enough seeding cells for the bon e tissue engineering. Methods The fibroblasts were isolated and purified from granu
lation of New Zealand rabbits, and they were incubated in the media offibronectin (FN) 10, 20, 40, 60 and 80 μg/ml, respectively, in the experimenta l grou ps 1- 5,but there was no FN in the control group. The markers for osteogenic
features were investigated by fibroblast morphogenesis,calcium nodules formationratios,labeling of tetracycline fluorescence, labeling of 3H-TdR, determination of o steocaline, and labeling of 3H-proline within 2 weeks. Results The morphologic al changes of the fibroblasts were manifested as transference from a long spindle to a round or multiple form, shifted nucleus increased in number, confluenced and formed multilayered structure. There was a piling-up of calcium crystals that were gradually merged into foggy substances. The foggy substances increased and formed
nodules. The calcium nodules formation ratios were as follows: 15.35%± 3.45%in the control group, and 53.73%± 9.49%, 75.21%± 9.80%, 98.34%± 15.2 0%, 61.83%± 10.04%, and 45.11%± 8.70% in the experimental groups 1.5 ,respectively. There was a significant difference between the control group and the 5 experimental groups at 14 days (P lt;0.05), and a significant differenc e be tween the experimental group 3 and the other experimental groups at 14 days (P lt;0.05). The histochemical study on the nodules with the specific labeling of tet racycline fluorescence indicated that the nodules were composed of new bones. Conclusion Fibronectin can stimulate the fibroblast to prolifer
ate, secrete osteocaline, and synthesize collagen fibrils. Fibronectin, in an optimal dose of 40 -60 μg/ml, is capable of inducing the fibroblast to form the bone.
Citation: L I Xiulan,ZHANG Yang,SHI Yijian,et al.. FIBRONECTIN OSTEOGENIC PHENOTYPICAL EXPPESSION BY FIBROBLASTS IN VITRO. Chinese Journal of Reparative and Reconstructive Surgery, 2006, 20(2): 107-111. doi: Copy