Younis Luay Thanoon, Abu Hassan Mohamed Ibrahim, Taiyeb Ali Tara Bai, Bustami Tommy Julianto
Faculty of Dentistry, Universiti Teknologi MARA, Sungai Buloh 47000, Malaysia.
Faculty of Dentistry, Universiti Teknologi MARA, MAHSA University, Jenjarom 42610, Malaysia.
Asian J Pharm Sci. 2018 Jul;13(4):317-325. doi: 10.1016/j.ajps.2017.12.003. Epub 2017 Dec 8.
This study was designed to investigate the effect of 3D TECA hydrogel on the inflammatory-induced senescence marker, and to assess the influence of the gel on the periodontal ligament fibroblasts (PDLFs) migration in wound healing . PDLFs were cultured with 20 ng/ml TNF-α to induce inflammation in the presence and absence of 50 µM 3D TECA gel for 14 d. The gel effect on the senescence maker secretory associated-β-galactosidase (SA-β-gal) activity was measured by a histochemical staining. Chromatin condensation and DNA synthesis of the cells were assessed by 4',6-diamidino-2-phenylindole and 5-ethynyl-2'-deoxyuridine fluorescent staining respectively. For evaluating fibroblasts migration, scratch wound healing assay and Pro-Plus Imaging software were used. The activity of senescence marker, SA-β-gal, was positive in the samples with TNF-α-induced inflammation. SA-β-gal percentage is suppressed (>65%, < 0.05) in the treated cells with TECA gel as compared to the non-treated cells. Chromatin foci were obvious in the non-treated samples. DNA synthesis was markedly recognized by the fluorescent staining in the treated compared to non-treated cultures. Scratch wound test indicated that the cells migration rate was significantly higher (14.9 µm/h, < 0.05) in the treated versus (11 µm/h) for control PDLFs. The new formula of 3D TECA suppresses the inflammatory-mediated cellular senescence and enhanced fibroblasts proliferation and migration. Therefore, 3D TECA may be used as an adjunct to accelerate repair and healing of periodontal tissues.
本研究旨在探讨3D TECA水凝胶对炎症诱导的衰老标志物的影响,并评估该凝胶对伤口愈合过程中牙周膜成纤维细胞(PDLFs)迁移的影响。在有和没有50µM 3D TECA凝胶存在的情况下,将PDLFs与20 ng/ml肿瘤坏死因子-α(TNF-α)一起培养14天以诱导炎症。通过组织化学染色测量凝胶对衰老标志物分泌相关β-半乳糖苷酶(SA-β-gal)活性的影响。分别通过4',6-二脒基-2-苯基吲哚和5-乙炔基-2'-脱氧尿苷荧光染色评估细胞的染色质凝聚和DNA合成。为了评估成纤维细胞的迁移,使用了划痕伤口愈合试验和Pro-Plus成像软件。衰老标志物SA-β-gal的活性在TNF-α诱导炎症的样本中呈阳性。与未处理的细胞相比,用TECA凝胶处理的细胞中SA-β-gal百分比受到抑制(>65%,P<0.05)。在未处理的样本中染色质病灶明显。与未处理的培养物相比,经荧光染色在处理的样本中明显识别出DNA合成。划痕伤口试验表明,处理组细胞迁移率(14.9µm/h,P<0.05)明显高于对照组PDLFs(11µm/h)。3D TECA的新配方可抑制炎症介导的细胞衰老,并增强成纤维细胞的增殖和迁移。因此,3D TECA可作为一种辅助手段来加速牙周组织的修复和愈合。