摘要
The development of the activated cellular bony implant, in light of the principle on tissue engineering, has brought about a new era to the fields of dental maxillofacial implantation. The present study separated the osteoblast like cells from human alveolar bone and seeded them into 3 types of biodegradable scaffold to form the complexes and then evaluated their osteogenic activities in vitro, in order to acquire experimental data that are essential to future clinical practice of this new type of therapeutical procedure in oral and maxillofacial surgery. Material and methods: Human alveolar bone origin cells were separated from alveolar bone around the third impacted teeth of 3 patients by enzyme digestion and went on cultures with α MEM containing β glycerophosphate and Dexamethasone at 5% CO2 ,37℃ for 21 28 days. Confirmed osteoblasts like cells were then seeded onto 3 types of degradable biomaterials of polyglycolic acid scaffold, collagen sponge, and L lactic acid/ε caprolactone to form cell matrices complexes. The 3 types of complex were continued to culture for 21 28 days in vitro at the same conditions with the single layer cultured cells. The cell proliferation, morphological changes, ALPase activity and mineral nodules formation on scaffolds were measured and observed at 3 days intervals to evaluate the affinities & the osteogenic activities of the human alveolar osteoblast like cells in the 3 different complexes. Result and discussion: The results indicated that the cultured human alveolar bone origin cells from 3 patients could successfully express the osteoblasts phenotype in single layered culturing in vitro after stimulated by β glycerophosphate and Dexamethasone. It has been shown that the cultured osteoblast like cells seeded on PGAS matrix had the highest attachmental, proliferative and osteogenic activities, suggesting a good bio affinity between the human alveolar osteoblast like cells and the PGAS matrix. The statistical analysis (ANOVA) showed that there were significant differences between PGAS osteoblasts complex and CLGS or LACT complexes on osteogenic activities. (P<0.05). It was also noticed that cultured human alveolar osteoblasts seeded in biodegradable materials had a delayed peak period on cell proliferation and PLAase production ,suggesting the osteoblasts seeded on scaffolds need a period of time to adjust themselves before they can normally proliferate and expres their phenotypes. Conclusion: PGAS osteoblasts complex is worth to be further developed into a tissue engineered cellular artificial bony implant for reconstructing the oral maxillofacial bony defects in a more effective way in the future.
The development of the activated cellular bony implant, in light of the principle on tissue engineering, has brought about a new era to the fields of dental maxillofacial implantation. The present study separated the osteoblast like cells from human alveolar bone and seeded them into 3 types of biodegradable scaffold to form the complexes and then evaluated their osteogenic activities in vitro, in order to acquire experimental data that are essential to future clinical practice of this new type of therapeutical procedure in oral and maxillofacial surgery. Material and methods: Human alveolar bone origin cells were separated from alveolar bone around the third impacted teeth of 3 patients by enzyme digestion and went on cultures with α MEM containing β glycerophosphate and Dexamethasone at 5% CO2 ,37℃ for 21 28 days. Confirmed osteoblasts like cells were then seeded onto 3 types of degradable biomaterials of polyglycolic acid scaffold, collagen sponge, and L lactic acid/ε caprolactone to form cell matrices complexes. The 3 types of complex were continued to culture for 21 28 days in vitro at the same conditions with the single layer cultured cells. The cell proliferation, morphological changes, ALPase activity and mineral nodules formation on scaffolds were measured and observed at 3 days intervals to evaluate the affinities & the osteogenic activities of the human alveolar osteoblast like cells in the 3 different complexes. Result and discussion: The results indicated that the cultured human alveolar bone origin cells from 3 patients could successfully express the osteoblasts phenotype in single layered culturing in vitro after stimulated by β glycerophosphate and Dexamethasone. It has been shown that the cultured osteoblast like cells seeded on PGAS matrix had the highest attachmental, proliferative and osteogenic activities, suggesting a good bio affinity between the human alveolar osteoblast like cells and the PGAS matrix. The statistical analysis (ANOVA) showed that there were significant differences between PGAS osteoblasts complex and CLGS or LACT complexes on osteogenic activities. (P<0.05). It was also noticed that cultured human alveolar osteoblasts seeded in biodegradable materials had a delayed peak period on cell proliferation and PLAase production ,suggesting the osteoblasts seeded on scaffolds need a period of time to adjust themselves before they can normally proliferate and expres their phenotypes. Conclusion: PGAS osteoblasts complex is worth to be further developed into a tissue engineered cellular artificial bony implant for reconstructing the oral maxillofacial bony defects in a more effective way in the future.
