摘要
目的 应用组织工程的原理和方法 ,体外构建人细胞型颌骨植骨材料并对其生物亲和性和成骨活性进行评价 ,探讨临床采用此类植骨材料修复口腔颌面骨缺损的可行性。方法 取 3名阻生齿患者的健康齿槽骨组织 ,经酶消化法进行齿槽骨细胞的分离 ,在 5 %CO2 、37℃的条件下用含成骨细胞分化诱导剂 β 磷酸甘油和地塞米松的α MEM(α minimumessentialmedium ,极限必需培养基 )培养液进行细胞的培养 ,以诱导其向成骨细胞分化。确定为成骨样细胞后 ,以 5× 10 4 /ml的密度分别播种在 3种可吸收性支架材料聚羟基乙酸 (PGAS)、胶原海绵 (CLGS)和L 乳酸 /ε 己酸内酯 (LACT)上 ,形成齿槽骨成骨样细胞 可吸收支架复合体。 3种复合体在体外同等条件下继续培养 3～ 4周 ,定期观测细胞在支架中的增殖速率、形态学变化、碱性磷酸酶活性及矿化结节形成等成骨活性指标的变化。结果 自 3人下颌齿槽骨分离培养的齿槽骨细胞在加入 β 磷酸甘油和地塞米松刺激的体外培养条件下 ,均可成功表达成骨细胞的表现型 ,与同源成纤维细胞相比 ,差异有显著性 (P <0 0 5 )。该齿槽骨成骨样细胞与 3种可吸收性支架材料分别构建成 3种细胞型骨种植材料后 ,PGAS中的细胞增殖、碱性磷酸酶活性和矿化结节形成量最多 ;统计学结果显示 ,其综合?
Objective To assembly human cellular alveolar bony implanting materials in vitro and evaluate their osteogenic activities. Methods Human alveolar bone cells were separated from alveolar bone around the third impacted teeth of 3 patients by enzyme digestion and cultured in α MEM containing β glycerophosphate and Dexamethasone at 5% CO 2, 37℃ for 21 28 days Confirmed osteoblasts like cells were then seeded onto 3 kinds of degradable biomaterials of polyglycolic acid scaffold, collagen sponge, and L lactic acid/ε caprolactone to form the cell scaffold complexes. The 3 types of complexes were continued to culture for 21 28 days at the same conditions. The cell proliferation, morphological changes, ALPase activity and mineral nodule formation on scaffolds were measured and observed at 3 days intervals. Results The results indicated that the cultured human alveolar bone origin cells from 3 patients could successfully express the osteoblasts phenotype in single layered culturing after stimulated by β glycerophosphate and Dexamethasone. The cultured osteoblast like cells seeded on PGAS matrix had the highest attachment, 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). Conclusion PGAS osteoblast complex is worth to be further developed into a tissue engineered cellular artificial bony implant for reconstructing the oral maxillofacial bony defects.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2001年第3期170-173,T001,共5页
Chinese Journal of Stomatology
基金
1998年度川医学奖学金资助项目
关键词
组织工程
人齿槽骨
植骨材料
颌面骨缺损
Tissue engineering
Human alveolar bone
Bony implanting materials
