Filho Guenther Schuldt, Caballé-Serrano Jordi, Sawada Kosaku, Bosshardt Dieter D, Bianchini Marco Aurélio, Buser Daniel, Gruber Reinhard
*Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, Bern, Switzerland.
†Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern.
J Periodontol. 2015 Jun;86(6):827-34. doi: 10.1902/jop.2015.140676. Epub 2015 Mar 19.
Demineralized bone matrix (DBM) is used for the treatment of osseous defects. Conditioned medium from native bone chips can activate transforming growth factor (TGF)-β signaling in mesenchymal cells. The aim of this study is to determine whether processing of native bone into DBM affects the activity of the conditioned medium.
Porcine cortical bone blocks were subjected to defatting, different concentrations of hydrochloric acid, and various temperatures. DBM was lyophilized, ground, and placed into culture medium. Human gingiva and periodontal fibroblasts were exposed to the respective conditioned medium obtained from DBM (DBCM). Changes in the expression of TGF-β target genes were determined.
DBCM altered the expression of TGF-β target genes (e.g., adrenomedullin, pentraxin 3, KN motif and ankyrin repeat domains 4, interleukin 11, NADPH oxidase 4, and BTB [POZ] domain containing 11) by at least five-fold. The response was observed in fibroblasts from both sources. Defatting lowered the activity of DBCM. The TGF-β receptor type I kinase inhibitor SB431542 [4-(4-(benzo[d][1,3]dioxol-5-yl)-5-(pyridin-2-yl)-1H-imidazol-2-yl)benzamide] but not the inhibitor of bone morphogenetic protein receptor dorsomorphin, blocked the effects of DBCM on gene expression. Moreover, conditioned medium obtained from commercial human DBM modulated the expression of TGF-β target genes.
The findings suggest that the DBCM can activate TGF-β signaling in oral fibroblasts.
脱矿骨基质（DBM）用于治疗骨缺损。天然骨碎片的条件培养基可激活间充质细胞中的转化生长因子（TGF）-β信号通路。本研究的目的是确定将天然骨加工成DBM是否会影响条件培养基的活性。
对猪皮质骨块进行脱脂、不同浓度盐酸处理及不同温度处理。将DBM冻干、研磨后置于培养基中。将人牙龈和成纤维细胞暴露于从DBM获得的相应条件培养基（DBCM）中。测定TGF-β靶基因表达的变化。
DBCM使TGF-β靶基因（如肾上腺髓质素、五聚素3、KN基序和锚蛋白重复结构域4、白细胞介素11、NADPH氧化酶4和含BTB[POZ]结构域1蛋白）的表达至少增加了五倍。在两种来源的成纤维细胞中均观察到该反应。脱脂降低了DBCM的活性。I型TGF-β受体激酶抑制剂SB431542[4-(4-(苯并[d][1,3]二氧杂环戊烯-5-基)-5-(吡啶-2-基)-1H-咪唑-2-基)苯甲酰胺]而非骨形态发生蛋白受体抑制剂多穗柯碱，可阻断DBCM对基因表达的影响。此外，从市售人DBM获得的条件培养基可调节TGF-β靶基因的表达。
研究结果表明，DBCM可激活口腔成纤维细胞中的TGF-β信号通路。
