不同来源成骨细胞构建组织工程骨之分子生物学评价

Expressions of Special Genes on Tissue Engineered Bone Contructed by Different Original Osteoblasts

目的:探讨骨膜来源成骨细胞(POB)和盖骨来源成骨细胞(COB)在构建组织工程骨中的黏附、增殖和分化能力。方法:研究分别选用人胚POB和COB为种子细胞,接种于生物衍生骨上,经过10天的体外培养,提取总RNA,经过逆转录成cDNA,采用实时定量PCR对目的基因成骨细胞特异转录因子(Cbfa1)、成骨细胞特异基因(osterix)、I型胶原(ColI)、骨钙素(osteocalcin,OC)以及整合素Integrinα5β1读取扩增循环数(cycle threshold,Ct)。结果:在组织工程骨中,POB和COB对ColI、OC、Cbfa1以及os-terix的表达没有统计学意义的差别,Integrinα5β1的表达可见POB明显高于COB。结论:POB与COB在与生物衍生材料接触后,仍可保持终末的分化性状,三维孔隙结构促进成骨细胞迅速而活跃的完成增殖期。POB对生物衍生骨的粘附能力优于COB。

Objective： The selection of seed cell is not only considered about its biology characters singly cultured in vitro but also based on understanding the interaction between the cell and biomaterials. This research tried to explore the ability of different original fetal osteoblasts, including periosteum-origin （POB） and cranium-origin（COB）, about adhesion, proliferation and differentiation. Methods： The fetal osteoblasts including periosteum-origin and cranium-origin, used as seed cells, constructed tissue engineered bone with the bioderived materials, and were cultured for 10 days in vitro. Real-time PCR analysis indicated the expressions of Cbfa-1, osterix, collagen I, osteocalcin and Integrinα5β1 mRNA. Results In tissue engineered bone, no statistical difference was found between POB and COB when they expressed Coil, OC, Cbfal and osterix mRNA, while POB expressed Integrinα5β1 more highly than COB. Both POB and COB could keep their phenotype after being seeded with bio-derived scaffolds. The differentiation of COB was better than POB and bio-derived bone supply great space which was benefit to COB to keep phenotype. Both Coil and OC were expressed lowerly in POB which indicated that POB obtained more proliferative ability on bio-derived bones and the three-dimensional pore construction could promote osteoblasts to complete proliferation quickly. Conclusions： Both POB and COB could keep their phenotype after being seeded with bio-derived scaffolds, and the three-dimensional pore construction could promote osteoblasts to complete proliferation quickly. POB shows stronger ability of adhesion than COB.