Iwai Yasutomo, Matsuda Yoshifumi, Nakatsuka Michiko, Mikami Yutaka, Kumabe Shunji
Department of Oral Anatomy, Osaka Dental University 8-1, Kuzuhahanazono-cho, Hirakata-shi, Osaka 573-1121, Japan.
Okajimas Folia Anat Jpn. 2012 Feb;88(4):133-40. doi: 10.2535/ofaj.88.133.
HMS0014 cells were GBR-engineered to proliferate and differentiate into mature osteoblast(Ob)-like cells, which initiated hard tissue matrix deposition in both monolayer and PuraMatrix 3-D cultures. Subsequently, the osteogenesis initiated with attachment/adhesion of HMS0014 cells on either Titanium (Ti) or Ti alloy discs modified with osteoconductive/ osteoinductive surface textures/substrates (e.g., Disc-AO, Disc-HA, Disc-SPI) was histologically assessed. The results obtained were as follows: 1) The HMS0014 cells actively proliferated and differentiated into mature Obs to initiate mineralisation of the ECM since day 1 in both monolayer and 3-D cultures; mineralization was prominently progressed between day 7 and day 14 of cultures. 2) The SEM of 60-minute(min)s specimens demonstrated a loose distribution of proliferating spherical-to-polygonal (10 to 40 microm in diameter, avg.) cells with a bulging cell body sending out many minute filopodia and some lamellipodia to attach with the substrate in the concavities. 3) In the 180-min specimens, the cultured HMS0014 cells actively proliferated and spread into flat, large polygonal cells with prominent lamellipodia and dendritic filopodia (30 microm x 90 microm to 100 microm x 200 microm, approx.) to employ cell-to-substrate and intercellular attachments. 4) On the other hand, the present immunohistochemistry of the attached HMS0014 cells demonstrated the co-expression of F-actin (actin filaments of the cytoskeleton) and CD51 (aV integrin) in both the 60-min and 180-min specimens. We concluded that the present GBR method enhanced HMS0014 cells to initiate an osteogenesis process with a direct bone-to-substratum contact on Ti discs which were subject to different surface modifications.
将HMS0014细胞进行引导骨再生工程处理,使其增殖并分化为成熟的成骨细胞样细胞,这些细胞在单层培养和PuraMatrix三维培养中均开始进行硬组织基质沉积。随后,对HMS0014细胞附着/黏附于经骨传导性/骨诱导性表面纹理/底物修饰的钛(Ti)或钛合金圆盘(如圆盘AO、圆盘HA、圆盘SPI)上后启动的成骨过程进行了组织学评估。获得的结果如下:1)在单层培养和三维培养中,自第1天起,HMS0014细胞就积极增殖并分化为成熟的成骨细胞,启动细胞外基质矿化;矿化在培养的第7天至第14天显著进展。2)60分钟标本的扫描电子显微镜显示,增殖的球形至多边形细胞(直径平均为10至40微米)分布松散,细胞体鼓起,发出许多微小的丝状伪足和一些片状伪足,以附着于凹面中的底物。3)在180分钟标本中,培养的HMS0014细胞积极增殖并扩散为扁平的大多边形细胞,具有明显的片状伪足和树枝状丝状伪足(约30微米×90微米至100微米×200微米),以实现细胞与底物及细胞间的附着。4)另一方面,对附着的HMS0014细胞进行的免疫组织化学分析表明,在60分钟和180分钟标本中均存在F-肌动蛋白(细胞骨架的肌动蛋白丝)和CD51(αV整合素)的共表达。我们得出结论,当前的引导骨再生方法增强了HMS0014细胞在经过不同表面修饰的钛圆盘上通过直接骨与底物接触启动成骨过程的能力。