Division of Oral Bioengineering, Department of Tissue Regeneration and Reconstitution, Institute of Medicine and Dentistry, Niigata University, Niigata, Japan.
J Periodontol. 2010 Mar;81(3):420-7. doi: 10.1902/jop.2009.090523.
A superporous (85%) hydroxyapatite (HA) block was recently developed to improve osteoconductivity, but it was often not clinically successful when used to treat periodontal osseous defects. The primary purpose of this study is to develop a clinically applicable tissue-engineered bone substitute using this HA block and human alveolar periosteum-derived cells.
Commercially available superporous HA blocks were acid treated and subjected to a three-dimensional (3D) culture for periosteal cell cultivation. Cells in the pore regions of the treated HA block were observed on the fracture surface by scanning electron microscopy. After osteogenic induction, the cell-HA complexes were implanted subcutaneously in nude mice. Osteoid formation was histologically evaluated.
Acid treatment enlarged the interconnections among pores, resulting in the deep penetration of periosteal cells. Under these conditions, cells were maintained for >2 weeks without appreciable cell death in the deep pore regions of the HA block. The cell-HA complexes that received in vitro osteogenic induction formed osteoids in pore regions of the treated HA blocks in vivo. In contrast, most pore regions in the non-pretreated, cell-free HA blocks that were evaluated in vivo remained cell free.
Our findings suggest that an acid-treated HA block could function as a better scaffold for the 3D high-density culture of human periosteal cells in vitro, and this cell-HA complex had significant osteogenic potential at the site of implantation in vivo. Compared with the cell-free HA block, our cell-HA complex using periosteal cells, which are the most accessible for clinical periodontists, showed promising results as a bone substitute in periodontal regenerative therapy.
最近开发了一种超高孔隙率(85%)的羟基磷灰石(HA)块,以提高成骨能力,但在用于治疗牙周骨缺损时,其临床效果往往不佳。本研究的主要目的是利用这种 HA 块和人牙槽骨膜来源的细胞开发一种临床可用的组织工程骨替代物。
对市售的超高孔隙率 HA 块进行酸处理,并进行三维(3D)培养以培养骨膜细胞。通过扫描电子显微镜观察处理后的 HA 块的断裂表面上的孔区中的细胞。成骨诱导后,将细胞-HA 复合物皮下植入裸鼠。通过组织学评估矿化骨的形成。
酸处理扩大了孔之间的连接,使骨膜细胞深入渗透。在这些条件下,细胞在 HA 块深部孔区保持>2 周而没有明显的细胞死亡。接受体外成骨诱导的细胞-HA 复合物在体内处理后的 HA 块的孔区中形成类骨质。相比之下,体内评估的未经预处理、无细胞的 HA 块的大多数孔区仍然没有细胞。
我们的研究结果表明,酸处理的 HA 块可作为体外人骨膜细胞 3D 高密度培养的更好支架,并且这种细胞-HA 复合物在体内植入部位具有显著的成骨潜力。与无细胞的 HA 块相比,我们使用最容易被临床牙周病医生获得的骨膜细胞的细胞-HA 复合物作为牙周再生治疗中的骨替代物显示出有希望的结果。