Lan Xu, Yang Zhi-ming, Ge Bao-feng, Liu Xue-mei
Division of Stem Cell and Tissue Engineering, Key Laboratory of Biotherapy of Human Diseases of Ministry of Education, West China Hospital, Sichuan University, Chengdu 610041, China.
Chin J Traumatol. 2005 Apr;8(2):86-90.
To study the effect of bio-derived bones, as substitutes of autogenous bone grafts and demineralized cadaver bones, on the attachment, spreading and proliferation of isolated osteoblasts.
Osteoblasts were isolated from the calvaria of a fetal rabbit through sequential collagenase digestion. In the attachment study, the osteoblasts labeled with 3H-leucine were incubated with the bio-derived bone materials in sterile microcentrifugable tubes for 15, 90 and 180 minutes, and 24 hours, respectively. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. In the proliferation study, the osteoblasts were cultured with the bio-derived bone materials for 24 hours and 3H-thymidine was added during the last 2 hours of the incubation. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. Osteoblasts were seeded on the bone graft materials for 60 or 120 minutes, 24 or 48 hours, and 3 or 7 days, then the co-culture was processed for scanning electron microscopy to observe the interaction of osteoblasts and the bio-derived bone materials.
Osteoblasts attached to the bio-derived bone materials in a time-dependent manner. There were significantly (P<0.05) more attached cells after 180 minutes than after 15 and 90 minutes of incubations (P<0.05). Osteoblasts were proliferated in a large amount on the surface and in the materials. Osteoblasts seeded onto 100 mg bio-derived bones resulted in significantly (P<0.05) more measurable proliferation than those seeded onto 10 mg bones. Osteoblasts appeared round as they attached to the materials, then flattened and spread over with time passing.
Bio-derived bones can provide a good environment for the attachment and proliferation of osteoblasts.
研究生物衍生骨作为自体骨移植和脱矿质尸体骨替代品，对分离的成骨细胞附着、铺展和增殖的影响。
通过连续胶原酶消化从胎兔颅骨分离成骨细胞。在附着研究中，将用³H-亮氨酸标记的成骨细胞分别与生物衍生骨材料在无菌微量离心管中孵育15分钟、90分钟、180分钟和24小时。收集附着的细胞，用液体闪烁光谱法测量放射性。在增殖研究中，将成骨细胞与生物衍生骨材料培养24小时，并在孵育的最后2小时加入³H-胸腺嘧啶核苷。收集附着的细胞，用液体闪烁光谱法测量放射性。将成骨细胞接种在骨移植材料上60或120分钟、24或48小时以及3或7天，然后对共培养物进行扫描电子显微镜检查，以观察成骨细胞与生物衍生骨材料的相互作用。
成骨细胞以时间依赖性方式附着于生物衍生骨材料。孵育180分钟后的附着细胞明显多于15分钟和90分钟后的附着细胞（P<0.05）。成骨细胞在材料表面和材料中大量增殖。接种到100mg生物衍生骨上的成骨细胞比接种到10mg骨上的成骨细胞增殖量明显更多（P<0.05）。成骨细胞附着于材料时呈圆形，随后随着时间推移变扁平并铺展开来。
生物衍生骨可为成骨细胞的附着和增殖提供良好环境。
