Stephan E B, Jiang D, Lynch S, Bush P, Dziak R
Department of Restorative Dentistry, SUNY at Buffalo School of Dental Medicine, NY 14214, USA.
J Periodontol. 1999 Apr;70(4):364-9. doi: 10.1902/jop.1999.70.4.364.
It was the aim of these studies to examine the ability of an anorganic bovine bone matrix material as an alternative to autogenous bone grafts and demineralized cadaver bone to support the attachment, spreading, and proliferation of isolated osteoblastic cells.
Primary culture osteoblastic cells were isolated from neonatal rat calvaria by sequential collagenase digestion. In the attachment studies, cells which had been labeled with 3H-leucine were incubated with the matrix material in sterile microfuge tubes for 15, 90, or 180 minutes or 24 hours. The attached cells were released and the radioactivity measured by liquid scintillation spectrometry. In the proliferation experiments, the cells were cultured with the matrix material for 24 hours and 3H-thymidine was added during the last 2 hours of the incubation. The cells were released and the radioactivity measured by liquid scintillation spectrometry. Scanning electron microscopy (SEM) was employed to observe osteoblastic cell interaction with the anorganic bone matrix. In these studies the cells were seeded on the bone graft material, then the material was removed and processed for SEM after 30, 60 or 120 minutes, or 24 or 48 hours.
The cells attached to the matrix material in a time-dependent manner. There were significantly (P<0.05) more cells attached after 180 minutes than after the 15 and 90 minute incubations. The matrix material also supported proliferation of the attached osteoblastic cells. Cells seeded onto 100 mg of anorganic bovine bone resulted in significantly (P<0.05) more measurable proliferation than cells seeded onto 10 mg of material. The cells appeared to be round as they attached, then flatten and spread over time. There was also evidence of cellular processes extending into the pores of the material.
These results demonstrate that this anorganic bovine bone graft material is able to support the attachment and proliferation of osteoblastic cells.
这些研究的目的是检验一种无机牛骨基质材料替代自体骨移植和脱矿人骨来支持分离的成骨细胞附着、铺展和增殖的能力。
通过连续胶原酶消化从新生大鼠颅骨分离原代培养的成骨细胞。在附着研究中，用³H-亮氨酸标记的细胞在无菌微量离心管中与基质材料孵育15、90或180分钟或24小时。释放附着的细胞，通过液体闪烁光谱法测量放射性。在增殖实验中，细胞与基质材料培养24小时，并在孵育的最后2小时加入³H-胸腺嘧啶核苷。释放细胞，通过液体闪烁光谱法测量放射性。采用扫描电子显微镜（SEM）观察成骨细胞与无机骨基质的相互作用。在这些研究中，将细胞接种在骨移植材料上，然后在30、60或120分钟或24或48小时后取出材料并进行SEM处理。
细胞以时间依赖性方式附着于基质材料。孵育180分钟后附着的细胞明显（P<0.05）多于孵育15和90分钟后的细胞。基质材料也支持附着的成骨细胞增殖。接种到100mg无机牛骨上的细胞比接种到10mg材料上的细胞产生明显（P<0.05）更多的可测量增殖。细胞附着时呈圆形，然后随着时间推移变平并铺展。也有证据表明细胞突起延伸到材料的孔隙中。
这些结果表明这种无机牛骨移植材料能够支持成骨细胞的附着和增殖。
