de Souza A P, Gerlach R F, Line S R
Departamento de Morfologia, Faculdade de Odontologia de Piracicaba-UNICAMP, Av. Limeira, 901, 13414-018 Piracicaba SP, Brazil.
Dent Mater. 2000 Mar;16(2):103-8. doi: 10.1016/s0109-5641(99)00084-6.
The interaction between metal ions and the oral environment is a major subject matter in dental research. Matrix metalloproteinases (MMPs) have been implicated in several pathologic oral processes such as periodontal tissue destruction, root caries, tumour invasion and temporomandibular joint disorders. The aim of this work was to test the effect of Zn, Cu, Sn and Hg ions on the activity of the major gingival gelatinolytic MMPs.
Gingival explants were cultured overnight in DMEM and the activity of secreted enzymes was analyzed by gelatin zymography in buffers containing different metal ion concentrations. The major gelatinolytic proteinases present in the conditioned media were characterized as MMP-2 and MMP-9 by immunoprecipitation with specific antibodies. The eletrophoretic bands were scanned and the transmittance values were analyzed with the Sigmagel software (Sigma).
ZnSO4 was a strong inhibitor of MMP-2 (I50 = 15 microM) and MMP-9 (I50 = 40 microM), whereas CuSO4, HgSO4 and SnCl2 showed less efficient inhibition potential.
Our findings show that the activity of oral tissue MMPs may be modulated by metal ions present in the oral environment. Therefore, the accumulation of metals in connective tissue may interfere with the formation and resorption of the extracellular matrix components.
金属离子与口腔环境之间的相互作用是牙科研究中的一个主要课题。基质金属蛋白酶(MMPs)与多种口腔病理过程有关,如牙周组织破坏、根龋、肿瘤侵袭和颞下颌关节紊乱。这项工作的目的是测试锌、铜、锡和汞离子对主要牙龈明胶溶解MMPs活性的影响。
牙龈外植体在DMEM中培养过夜,通过明胶酶谱法在含有不同金属离子浓度的缓冲液中分析分泌酶的活性。通过用特异性抗体进行免疫沉淀,将条件培养基中存在的主要明胶溶解蛋白酶鉴定为MMP-2和MMP-9。扫描电泳条带,并用Sigmagel软件(Sigma)分析透光率值。
硫酸锌是MMP-2(I50 = 15 microM)和MMP-9(I50 = 40 microM)的强抑制剂,而硫酸铜、硫酸汞和氯化锡的抑制潜力较低。
我们的研究结果表明,口腔组织MMPs的活性可能受到口腔环境中存在的金属离子的调节。因此,结缔组织中金属的积累可能会干扰细胞外基质成分的形成和吸收。