Synthesis and biological evaluation of isoxazolyl-sulfonamides: A non-cytotoxic scaffold active against Trypanosoma cruzi, Leishmania amazonensis and Herpes Simplex Virus.

In this study we report the synthesis, characterization, biological evaluation, and druglikeness assessment of a series of 20 novel isoxazolyl-sulfonamides, obtained by a four-step synthetic route. The compounds had their activity against Trypanosoma cruzi, Leishmania amazonensis, Herpes Simplex Virus type 1 and cytotoxicity evaluated in phenotypic assays. All compounds have drug-like properties, showed low cytotoxicity and were promising regarding all other biological activities reported herein, especially the inhibitory activity against T. cruzi. The compounds 8 and 16 showed significant potency and selectivity against T. cruzi (GI50 = 14.3 µM, SI > 34.8 and GI50 = 11.6 µM, SI = 29.1, respectively). These values, close to the values of the reference drug benznidazole (GI50 = 10.2 µM), suggest that compounds 8 and 16 represent promising candidates for further pre-clinical development targeting Chagas disease.

Effect of temperature and seven storage media on human periodontal ligament fibroblast viability.

BACKGROUND/AIM: Natural resources, such as coconut water, propolis, and egg whites, have been examined as possible storage media for avulsed teeth. However, there is a lack of research focused on the efficacy of these three products together compared with Hank's balanced salt solution and milk. The aim of this study was to evaluate the capacity of seven storage media to maintain the viability of human periodontal ligament fibroblasts (PDLFs). MATERIAL AND METHODS: PDLFs were kept at 5°C and 20°C, in skimmed milk (SMilk), whole milk (WMilk), recently prepared Hank's balanced salt solution (HBSS), Save-A-Tooth® system's HBSS (Save), natural coconut water (Coconut), Propolis, and egg white (Egg) for 3, 6, 24, 48, 72, 96, and 120 h, through the analysis of tetrazolium salt-based colorimetric (MTT) assay. RESULTS: At 5°C, SMilk and WMilk were better than HBSS in maintaining cell viability, from 24 h onward. At 20°C, HBSS was the best storage medium at 96 and 120 h. At both temperatures, from 6 h onward, Coconut, Propolis and Egg were less effective than SMilk, WMilk, and HBSS. In general, the performance of Coconut, Propolis and Egg were not influenced by storage temperature. However, the lowest temperature undermined the effectiveness of HBSS from 24 h and favored SMilk and WMilk, from 96 and 48 h onward, respectively. Save and water were the worst storage media. CONCLUSION: SMilk was the best storage medium, followed by WMilk and HBSS. Coconut, Propolis, and Egg can be indicated for the conservation of PDLF up to 3 h. The lower temperature (5°C) undermined the effectiveness of HBSS and favored SMilk and WMilk.

Effect of Milk Renewal on Cell Viability In Vitro at Different Time Frames.

The purpose of this study was to evaluate if the renewal of milk as a storage medium, every 12, 24 and 48 h, is able to increase its ability to maintain human periodontal ligament fibroblasts (PDLF) viability over time. PDLF were soaked in Minimum Essential Medium at 37 °C (MEM-37) (positive control), tap water (Water) (negative control) and in skimmed milk (44 wells) at 5 °C and 20 °C. The skimmed milk was renewed every 12 h (Milk-12), 24 h (Milk-24) and 48 h (Milk-48) in 11 wells of each plate, and the milk in the remaining 11 wells of each plate was maintained in situ (not renewed milk) (NRM). After 24, 48, 72, 96 and 120 h, cell viability was determined by the tetrazolium salt-based colorimetric (MTT) assay. Data were statistically analyzed by Kruskal-Wallis, Scheffé and Mann-Whitney tests (a=5%). At 5 °C, only Milk-48 was significantly better than NRM. At 20 °C, NRM was more effective than Milk-12 and Milk-24 in all time periods. In relation to the temperature (5 °C or 20 °C), renewal of milk at 5 °C was better in maintaining cell viability than the renewal at 20 °C. In conclusion, the renewal of milk was able to increase its ability to maintain cell viability only when performed every 48 h in milk maintained at 5 °C.

Fibroblast Viability after Storage at 20 °C in Milk, Hank's Balanced Salt Solution and Coconut Water.

The objective of this study was to evaluate the effectiveness of various storage media at 20 °C in maintaining the viability of human periodontal ligament fibroblasts (HPLF) over time. HPLF were maintained at 20 °C in skim milk (SM), whole milk (WM), freshly prepared Hank's balanced salt solution (HBSS), Save-A-Tooth(r), natural coconut water (NCW), coconut water industrialized (ICW) and tap water (negative control) for 3, 6, 24, 48, 72, 96 and 120 h. Cells maintained in Minimal Essential Medium (MEM-37) at 37 °C served as a positive control. Cell viability was determined by MTT assay. Statistical analysis was performed by Kruskal-Wallis test and Scheffe test (α = 5%). From 24 h, NCW was significantly better in maintaining cell viability than all other tested storage media (p<0.05). SM and WM were significantly better than HBSS for up to 72 h. Save-A-Tooth(r) and ICW were the worst conservation storage media. In conclusion, the effectiveness of the tested storage media to maintain the viability of the periodontal ligament cells was as follows, in a descending order: NCW > MEM-37> SM and IM> HBSS> ICW > Save-A-Tooth(r)> tap water.

Fibroblast Viability after Storage at 20 °C in Milk, Hank's Balanced Salt Solution and Coconut Water

Abstract The objective of this study was to evaluate the effectiveness of various storage media at 20 °C in maintaining the viability of human periodontal ligament fibroblasts (HPLF) over time. HPLF were maintained at 20 °C in skim milk (SM), whole milk (WM), freshly prepared Hank's balanced salt solution (HBSS), Save-A-Tooth(r), natural coconut water (NCW), coconut water industrialized (ICW) and tap water (negative control) for 3, 6, 24, 48, 72, 96 and 120 h. Cells maintained in Minimal Essential Medium (MEM-37) at 37 °C served as a positive control. Cell viability was determined by MTT assay. Statistical analysis was performed by Kruskal-Wallis test and Scheffe test (α = 5%). From 24 h, NCW was significantly better in maintaining cell viability than all other tested storage media (p<0.05). SM and WM were significantly better than HBSS for up to 72 h. Save-A-Tooth(r) and ICW were the worst conservation storage media. In conclusion, the effectiveness of the tested storage media to maintain the viability of the periodontal ligament cells was as follows, in a descending order: NCW > MEM-37> SM and IM> HBSS> ICW > Save-A-Tooth(r)> tap water.

Resumo O objetivo deste estudo foi avaliar a efetividade de vários meios de conservação a 20 °C em manter a viabilidade de fibroblastos do ligamento periodontal humano (FLPH) ao longo do tempo. FLPH foram conservados a 20 °C em leite desnatado (LD), leite integral (LI), solução salina balanceada de Hank (HBSS) recém preparada, Save-A-Tooth(r) (Save), água de coco natural (ACN), água de coco industrializada (ACI) e água de torneira (água - controle negativo) por 3, 6, 24, 48, 72, 96 e 120 h. Células conservadas em Meio Essencial Mínimo (MEM-37) a 37 °C serviram como controle-positivo. A viabilidade celular foi determinada pelo ensaio MTT. A análise estatística dos dados foi realizada pelos testes Kruskal-Wallis e Scheffé (α=5%). A partir de 24 h, ACN foi significantemente melhor em manter a viabilidade celular do que todos os outros meios testados (p<0,05). LD e LI foram significantemente melhores do que a HBSS por até 72 h. Save e ACI foram os piores meios de conservação. Concluindo, a efetividade dos meios de conservação testados em manter a viabilidade das células do ligamento periodontal foi a seguinte em ordem decrescente: ACN > MEM-37 > LD e LI > HBSS > ACI > Save > água.