Zerbo Ilara R, Bronckers Antonius L J J, de Lange Gert, Burger Elisabeth H
Department of Oral Cell Biology, ACTA, Van der Boechorststraat 7, 1081BT Amsterdam, The Netherlands.
Biomaterials. 2005 Apr;26(12):1445-51. doi: 10.1016/j.biomaterials.2004.05.003.
We and others have shown earlier that porous beta-tricalcium phosphate (TCP) (Cerasorb) can be used in patients to augment the maxillary sinus floor prior to placement of oral dental implants. To better understand the transformation of TCP particles into bone tissue, we analyse here the appearance of cells with osteogenic or osteoclastic potential in relation to these particles. In biopsies taken at 6 months after sinus floor augmentation we observed bone growth into the TCP particles but also replacement by soft connective tissue. To identify possible osteoprogenitor cells in this tissue, histological sections were immunostained with an antibody to Runx2/Cbfa1, an essential and early transcription factor for osteoblast differentiation. The osteogenic potential of cells was further confirmed by immunostaining for bone sialoprotein (BSP) and osteopontin (OPN). Other sections were stained for Tartrate Resistant Acid Phosphatase (TRAP) activity to identify cells with osteoclastic capacity. Runx2/Cbfa1 positive connective tissue cells were found in abundance throughout and around the TCP particles, even at a distance of several millimetres from the maxillary bone surface. About 95% of the cells found within TCP particles stained positive for Runx2/Cbfa1. Fewer cells stained positive for BSP and OPN, suggesting more mature osteoblastic properties. Mono- and binucleate TRAP-positive cells, but no multinucleate TRAP-positive osteoclasts, were found in the soft tissue infiltrating the TCP and at the surface of the TCP particles. Both the Runx2/Cbfa1 positive and the TRAP-positive cells decreased apically with increasing vertical distance from the maxillary bone surface. This data suggests that the TCP particles attract osteoprogenitor cells that migrate into the interconnecting micropores of the bone substitute material by 6 months. The lack of large multinucleate TRAP positive cells suggests that resorption of the TCP material by osteoclasts plays only a minor role in its replacement by bone. Chemical dissolution, possibly favoured by a high cell metabolism in the particles, seems the predominant cause of TCP degradation. The abundance of Runx2/Cbfa1 positive cells would indicate that with a greater time of healing there will be further bone deposition into these particles.
我们和其他研究人员之前已经表明,多孔β-磷酸三钙(TCP)(Cerasorb)可用于患者,在植入口腔种植牙之前提升上颌窦底。为了更好地理解TCP颗粒向骨组织的转化过程,我们在此分析了具有成骨或破骨潜能的细胞与这些颗粒的关系。在窦底提升6个月后采集的活检组织中,我们观察到骨长入TCP颗粒,但也有被软结缔组织替代的情况。为了识别该组织中可能的骨祖细胞,组织学切片用抗Runx2/Cbfa1抗体进行免疫染色,Runx2/Cbfa1是成骨细胞分化所必需的早期转录因子。通过对骨唾液蛋白(BSP)和骨桥蛋白(OPN)进行免疫染色,进一步证实了细胞的成骨潜能。其他切片用抗酒石酸酸性磷酸酶(TRAP)活性染色,以识别具有破骨能力的细胞。在整个TCP颗粒及其周围都大量发现了Runx2/Cbfa1阳性结缔组织细胞,甚至在距离上颌骨表面几毫米处也有。在TCP颗粒内发现的细胞中,约95%对Runx2/Cbfa1染色呈阳性。对BSP和OPN染色呈阳性的细胞较少,表明成骨细胞特性更成熟。在浸润TCP的软组织和TCP颗粒表面发现了单核和双核TRAP阳性细胞,但没有多核TRAP阳性破骨细胞。Runx2/Cbfa1阳性细胞和TRAP阳性细胞均随着距上颌骨表面垂直距离的增加而在根尖方向减少。该数据表明,到6个月时,TCP颗粒吸引骨祖细胞迁移到骨替代材料的相互连通的微孔中。缺乏大型多核TRAP阳性细胞表明,破骨细胞对TCP材料的吸收在其被骨替代过程中仅起次要作用。化学溶解,可能因颗粒中高细胞代谢而更易发生,似乎是TCP降解的主要原因。Runx2/Cbfa1阳性细胞的大量存在表明,随着愈合时间延长,这些颗粒中将有更多的骨沉积。