Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts.
Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts.
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DOI：
10.1902/jop.2000.71.4.586
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发表时间：
2000-04
影响因子：
4.3
通讯作者：
Z. Schwartz;Z. Schwartz;C. H. Lohmann;C. H. Lohmann;M. Wieland;David L. Cochran;David D Dean;M. Textor;L. F. Bonewald;B. Boyan
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文献类型：
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作者：
Z. Schwartz;Z. Schwartz;C. H. Lohmann;C. H. Lohmann;M. Wieland;David L. Cochran;David D Dean;M. Textor;L. F. Bonewald;B. Boyan
种植体表面粗糙度和化学成分，以及其他因素，影响成骨细胞在种植体附近形成骨的能力。同样的原理也适用于牙根，一些报告表明，牙根表面的修饰可以改善牙周器官的修复。这些表面修饰技术中最常见的包括用柠檬酸或四环素去除矿化以暴露胶原原纤维。此外，在正常骨重塑过程中，破骨细胞使细胞外基质脱矿，留下吸收坑和暴露的胶原原纤维。本研究比较了不同牙本质表面制备技术对成骨细胞的影响。方法抹香鲸牙本质切片机械抛光，表面用四环素-盐酸（TCN）处理，或与小鼠骨髓细胞培养形成表面破骨细胞（OC）吸收坑或不处理。采用轮廓术、x射线光电子能谱（XPS）和扫描电子显微镜（SEM）对3种不同牙本质表面进行评价。在3种不同表面培养MG63成骨样细胞，比较牙本质表面制备技术对MG63细胞增殖（细胞数量）、分化（分离细胞和细胞层裂解物碱性磷酸酶特异性活性、骨钙素生成）、局部因子生成(转化生长因子（TGF)- β 1和前列腺素E2 (PGE2)）的影响。结果磨光和TCN表面光滑，Ra值相当，而OC表面由于吸收坑（占表面的3.7%）而略粗糙。XPS测量表明，TCN处理降低了表面Ca和P的含量，表明它已经溶解了矿物。破骨细胞吸收也降低了钙和磷含量，但程度较轻。MG63细胞在抛光牙本质和组织培养聚苯乙烯上的增殖相当。相比之下，在TCN和oc处理过的表面上生长的细胞增殖能力增强。表面处理对细胞碱性磷酸酶活性没有影响，但在TCN和oc处理的表面，细胞层裂解物的活性增加。所有牙本质表面的骨钙素产生均减少，但tcn处理表面的骨钙素减少幅度最大。tgf - β 1和PGE2的生成在处理过的表面都增加了。在tcn处理的牙本质上培养的所有效果都最大。结论：牙本质表面脱矿促进了成骨细胞的增殖和早期分化，如碱性磷酸酶和自分泌介质如PGE2和tgf - β 1的产生。然而，后期分化事件如骨钙素的产生减少。破骨细胞介导的骨吸收引起类似的反应；不到4%的牙本质表面导致了大约75%的TCN治疗引起的反应。这些观察结果表明，在设计促进根表面附近骨和牙骨质形成的方法时，应更多地关注破骨细胞吸收的影响。
BACKGROUND Implant surface roughness and chemical composition, as well as other factors, affect the ability of osteogenic cells to form bone adjacent to an implant. The same principles may also apply to the tooth root and some reports have shown that surface modification of the root may lead to improved restoration of the periodontal apparatus. The most common of these surface modification techniques involves demineralization with citric acid or treatment with tetracycline to expose collagen fibrils. In addition, during normal bone remodeling, osteoclasts demineralize the extracellular matrix, leaving resorption pits and exposed collagen fibrils. In this study, the effect of different dentin surface-preparation techniques on osteoblasts were compared. METHODS Slices of sperm whale dentin were mechanically polished and surfaces were treated with tetracycline-HCl (TCN) or were cultured with mouse bone marrow cells to create a surface with osteoclast (OC) resorption pits or left untreated. Profilometry, x-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to evaluate the 3 different dentin surfaces. MG63 osteoblast-like cells were cultured on the 3 different surfaces and the effect of dentin surface preparation technique on MG63 cell proliferation (cell number), differentiaton (alkaline phosphatase specific activity of isolated cells and cell layer lysates; osteocalcin production), and local factor production (transforming growth factor (TGF)-beta1 and prostaglandin E2 (PGE2) compared. RESULTS Profilometry showed the polished and TCN surfaces were smooth with comparable Ra values, whereas the OC surfaces were slightly rougher due to resorption pits which covered 3.7% of the surface. XPS measurements showed that TCN treatment reduced the Ca and P content of the surface, indicating that it had dissolved the mineral. Osteoclast-resorption also reduced the Ca and P content, but to a lesser extent. MG63 cell proliferation on polished dentin and tissue culture polystyrene was equivalent. In contrast, cells grown on the TCN- and OC-treated surfaces exhibited increased proliferation. No effect of surface treatment on cell alkaline phosphatase activity was observed, but activity in the cell layer lysates was increased on the TCN- and OC-treated surfaces. Osteocalcin production was reduced on all dentin surfaces, but the greatest reduction was found on the TCN-treated surface. Production of both TGF-beta1 and PGE2 was increased on the treated surfaces. All effects were greatest in cultures grown on the TCN-treated dentin. CONCLUSIONS These data indicate that demineralization of the dentin surface promotes proliferation of osteoblasts and early differentiation events like production of alkaline phosphatase and autocrine mediators such as PGE2 and TGF-beta1. However, later differentiation events like osteocalcin production are decreased. Osteoclast-mediated bone resorption elicits similar responses; less than 4% of the dentin surface resulted in approximately 75% of the response caused by TCN treatment. These observations suggest that greater attention should be paid to the effects of osteoclastic resorption in designing methods for enhancing bone and cementum formation adjacent to root surfaces.