研究目的
To evaluate the leishmanicidal activity of rose bengal B and its ester derivatives (RBMET and RBBUT) against Leishmania amazonensis using photodynamic therapy with LED light.
研究成果
RBMET and RBBUT are effective against L. amazonensis in PDT, with IC50 values comparable to reference drugs. Despite low selectivity index due to cytotoxicity, their topical application in PDT may still be viable for CL treatment, as it could destroy infected cells. Future studies should focus on absorption and toxicity in topical formulations.
研究不足
The photosensitizers RBMET and RBBUT showed high cytotoxicity to macrophages, leading to low selectivity index. This toxicity prevented evaluation of activity on intracellular amastigotes. The study was in vitro, and further in vivo studies are needed to assess topical use and toxicity.
1：Experimental Design and Method Selection:
The study used photodynamic therapy (PDT) with xanthene dyes (RB, RBMET, RBBUT) irradiated with LED light to assess leishmanicidal activity and cytotoxicity. Methods included assays on promastigotes and intracellular amastigotes, and cytotoxicity tests on macrophages.
2：Sample Selection and Data Sources:
Promastigotes of Leishmania amazonensis (strain MHOM/BR/1977/LTB0016) and J774.A1 macrophages were cultured. Peritoneal macrophages from BALB/c mice were also used for infection assays.
3：A1 macrophages were cultured. Peritoneal macrophages from BALB/c mice were also used for infection assays.
List of Experimental Equipment and Materials:
3. List of Experimental Equipment and Materials: LED illumination system (575 nm/1.66 mW/cm2), Varian-Cary Eclipse spectrofluorometer (Agilent Technologies, USA), Handheld Laser Power Meter (Edmund Optics Inc., USA), 96-well plates, RPMI 1640 medium (Gibco), fetal calf serum, penicillin G, streptomycin, XTT reduction kit, rose bengal B (Neon, S?o Paulo, Brazil), and its derivatives RBMET and RBBUT.
4：66 mW/cm2), Varian-Cary Eclipse spectrofluorometer (Agilent Technologies, USA), Handheld Laser Power Meter (Edmund Optics Inc., USA), 96-well plates, RPMI 1640 medium (Gibco), fetal calf serum, penicillin G, streptomycin, XTT reduction kit, rose bengal B (Neon, S?o Paulo, Brazil), and its derivatives RBMET and RBBUT.
Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Promastigotes and macrophages were incubated with photosensitizers for 30 minutes in the dark, then illuminated with LED for 30 minutes. Cell viability was measured after 24, 48, and 72 hours using XTT reduction method. Cytotoxicity was assessed similarly on macrophages.
5：Data Analysis Methods:
IC50 and CC50 values were calculated using linear regression. Statistical analysis was performed with Statistica 12.0 software, using Mann-Whitney U test with significance level of 5%.
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