Inaba Hiroaki, Tagashira Motoyuki, Kanda Tomomasa, Ohno Takashi, Kawai Shinji, Amano Atsuo
Department of Oral Frontier Biology, Osaka University Graduate School of Dentistry, Osaka, Japan.
J Periodontol. 2005 Dec;76(12):2223-9. doi: 10.1902/jop.2005.76.12.2223.
Enamel matrix derivative (EMD) is a tissue regenerative agent used clinically as an adjunct to periodontal surgery. It was previously demonstrated that Porphyromonas gingivalis, a periodontal pathogen, significantly diminished the efficacy of EMD with periodontal ligament (PDL) cells through the proteolytic actions of Arg- and Lys-gingipains (Rgp and Kgp). Thus, antiproteolytic supplements are considered clinically desirable for effective periodontal regenerative therapies. In the present study, we examined apple- (AP) and hop-polyphenols to determine their ability to protect EMD-stimulated PDL cells from P. gingivalis.
AP, apple condensed tannin (ACT), hop bract polyphenol (HBP), high and low molecular weight fractions of HBP (HMW-HBP and LMW-HBP), and epigallocatechin gallate (EGCg) were used. PDL cells were grown on EMD-coated dishes and infected with P. gingivalis, and cellular migration and proliferation were evaluated with an in vitro assay of wound healing assay in the presence or absence of the polyphenols.
Each polyphenol significantly enhanced the viability of PDL cells infected with P. gingivalis, whereas only EGCg demonstrated cytotoxicity. Further, all polyphenols significantly inhibited Rgp activity, with AP, ACT, and HBP more effective toward Kgp. P. gingivalis markedly diminished the migration and proliferation of EMD-stimulated PDL cells, whereas the addition of AP, ACT, HBP, and HMW-HBP significantly protected the cells from bacterial cytotoxicity. In contrast, EGCg and LMW-HBP did not show protective effects.
These results suggest that AP, ACT, AP, HBP, and HMW-HBP protect EMD-stimulated PDL cells from P. gingivalis and may be therapeutically useful supplements for EMD therapy.
釉基质衍生物(EMD)是一种临床上用作牙周手术辅助剂的组织再生剂。先前的研究表明,牙周病原体牙龈卟啉单胞菌通过精氨酸和赖氨酸牙龈蛋白酶(Rgp和Kgp)的蛋白水解作用,显著降低了EMD对牙周膜(PDL)细胞的疗效。因此,抗蛋白水解补充剂被认为是有效的牙周再生治疗的临床理想选择。在本研究中,我们检测了苹果多酚(AP)和啤酒花多酚,以确定它们保护EMD刺激的PDL细胞免受牙龈卟啉单胞菌侵害的能力。
使用了AP、苹果缩合单宁(ACT)、啤酒花花苞多酚(HBP)、HBP的高分子量和低分子量级分(HMW-HBP和LMW-HBP)以及表没食子儿茶素没食子酸酯(EGCg)。将PDL细胞接种在涂有EMD的培养皿上,并用牙龈卟啉单胞菌感染,在有或没有多酚的情况下,通过体外伤口愈合试验评估细胞迁移和增殖。
每种多酚都显著提高了感染牙龈卟啉单胞菌的PDL细胞的活力,而只有EGCg表现出细胞毒性。此外,所有多酚都显著抑制Rgp活性,AP、ACT和HBP对Kgp的抑制作用更有效。牙龈卟啉单胞菌显著降低了EMD刺激的PDL细胞的迁移和增殖,而添加AP、ACT、HBP和HMW-HBP可显著保护细胞免受细菌细胞毒性的影响。相比之下,EGCg和LMW-HBP没有显示出保护作用。
这些结果表明,AP、ACT、AP、HBP和HMW-HBP可保护EMD刺激的PDL细胞免受牙龈卟啉单胞菌的侵害,可能是EMD治疗中有治疗作用的补充剂。