Teixeira Karina I R, Cortés Maria E, Santos Robson A S, Oliveira Flávio, Sinisterra Ruben D
Department of Restorative Dentistry, Universidade Federal de Minas Gerais, Av Antonio Carlos, 6627, CEP 31270-901 Belo Horizonte, MG, Brazil.
Department of Restorative Dentistry, Universidade Federal de Minas Gerais, Av Antonio Carlos, 6627, CEP 31270-901 Belo Horizonte, MG, Brazil and National Institute of Science and Technology in Nano-Biopharmaceutical, Belo Horizonte, MG, 31270901, Brazil.
Biointerphases. 2016 Dec 1;11(4):04B307. doi: 10.1116/1.4968880.
The aim of this study was to determine the physical properties and antimicrobial and antiproliferative effects of the KR12 peptide complexed with 2-hydroxypropyl-β-cyclodextrin (Hp-βCd) in vitro. The KR12:Hp-βCd composition was evaluated for particle size and its zeta (ζ)-potential in the presence and absence of cells. Antimicrobial activity against Streptococcus mutans, Actinobacillus actinomycetemcomitans, and Porphyromonas gingivalis for the peptide alone or associated was evaluated by minimal inhibitory concentration. The cytotoxicity of the peptide and composition toward fibroblasts, Caco-2 cells, and A431 cells was determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide; thiazolyl blue assay and hemolysis assay. Membrane integrity was analyzed by the lactate dehydrogenase assay. KR12:Hp-βCd decreased the peptide concentration required for the antimicrobial effect. Moreover, this composition was able to modify cell surface parameters, such as ζ-potential, and alter the degree of hemolysis induced by KR12. However, the KR12:Hp-βCd and KR12 alone alter the zeta potential of cells to a similar extent, suggesting a similar level of membrane interaction. The peptide alone inhibited the proliferation of Caco-2 and A431 cells more efficiently than KR12:Hp-βCd (p < 0.001), but did not show significant cytotoxic effects via the dehydrogenase lactate assay. Both substances were effective in inhibiting the growth of odontopathogenic bacteria, as well as inhibiting Caco-2 epithelial cells. These observations highlight the potential antimicrobial and antiproliferative effects of KR12 peptide alone or associated with Hp-βCd.
本研究的目的是在体外确定与2-羟丙基-β-环糊精（Hp-βCd）复合的KR12肽的物理性质、抗菌和抗增殖作用。评估了KR12:Hp-βCd组合物在有无细胞存在情况下的粒径及其zeta（ζ）电位。通过最小抑菌浓度评估单独或复合的该肽对变形链球菌、伴放线放线杆菌和牙龈卟啉单胞菌的抗菌活性。使用3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐；噻唑蓝法和溶血试验确定该肽及其组合物对成纤维细胞、Caco-2细胞和A431细胞的细胞毒性。通过乳酸脱氢酶试验分析膜完整性。KR12:Hp-βCd降低了抗菌作用所需的肽浓度。此外，该组合物能够改变细胞表面参数，如ζ电位，并改变KR12诱导的溶血程度。然而，单独的KR12和KR12:Hp-βCd对细胞zeta电位的改变程度相似，表明膜相互作用水平相似。单独的该肽比KR12:Hp-βCd更有效地抑制Caco-2和A431细胞的增殖（p < 0.001），但通过乳酸脱氢酶试验未显示出显著的细胞毒性作用。这两种物质在抑制致龋病原菌生长以及抑制Caco-2上皮细胞方面均有效。这些观察结果突出了单独的KR12肽或与Hp-βCd复合的KR12肽的潜在抗菌和抗增殖作用。
