Hematoporphyrin (Hematoporphyrin IX)

Cell lines

Preparation Method

5×10³ cells per well were seeded in 96-well plates. Following incubation for 12h at 37°C, the cells were treated with phosphate-buffered saline (PBS, control) or various concentrations of Hematoporphyrin (0 - 120nM) for 60min. Following light irradiation using an M8 Spectrum Power Energy meter at 625nm, 5mW/cm² for 60min and culturing for 22h at 37°C, 100μl MTT solution (5.0mg/ml) was then added to each well and incubated for 4h at 37°C, followed by removal of culture medium and the addition of dimethyl sulfoxide (100μl/well) to dissolve the formed formazan crystals. The absorbance value was determined using a microplate reader at a wavelength of 520nm. The relative cell viability was calculated by normalization to the control. The experiments were independently performed three times.

Reaction Conditions

Applications

Animal models

Preparation Method

Hematoporphyrin was dissolved in 150mmol/L buffered saline, and a 1mg/ml Hematoporphyrin solution was prepared.

Thirty BALB/c nude mice weighing between 15 and 18g were divided into three groups: normal control group (ten samples), model group (ten samples) and model with Hematoporphyrin group (ten samples).

The human gastric cancer cell line MCG-803 cells were cultured in a DMEM culture medium supplemented with 10% fetal calf serum, 50U/ml penicillin and 50μg/ml streptomycin. Suspensions of tumor cells MCG-803 (5 - 10×10⁶ viable cells/mouse) were implanted into the right dorsal portion of BALB/c nude mice. After 20 days, the inoculated tumors reached the diameter of about 0.8 - 1.0mm, the mice were killed and tumors were resected for establishing the animal model of gastric carcinoma in situ. The resected tumor tissues were sheared to small tumor tissues about 1mm³. The abdominal cavity of the 20 mice in model groups were opened by operation, the gastric wall was exposed and the small tumor tissues were then implanted into the gastric wall. After 20 days, when the diameter of the implanted tumor became about 0.4 - 0.6mm, the animals were prepared for optical coherence tomography (OCT) imaging.

24h before the OCT image detections, the normal control group, model group and model with hematoporphyrin group received 5ml/kg body weight NS, NS and Hematoporphyrin solution i.v. from vena caudalis, respectively. The animals were then still housed for 24h under specific pathogen-free conditions. On the second day, the mice were anesthetized using an intraperitioneal injection of a mixture of ketamine (100mg/ml) and xylazine (20mg/ml) 0.02ml/g body weight. The gastric body was exposed by surgical intervention, and the stomach mucosa tissues and implanted tumor tissues in nude mice are performed by OCT at a central wavelength of 1310nm. In addition, in the normal control group, the stomach mucosa tissues were locally injected a 0.02ml Hematoporphyrin solution, and were performed by OCT imaging after 30min.

Dosage form

Applications

References:
[1] Yuan SX, Li JL, Xu XK, et al. Underlying mechanism of the photodynamic activity of hematoporphyrin?induced apoptosis in U87 glioma cells. Int J Mol Med. 2018;41(4):2288-2296.
[2] Xiong H, Zeng C, Guo Z, et al. Potential ability of hematoporphyrin to enhance an optical coherence tomographic image of gastric cancer in vivo in mice. Phys Med Biol. 2008;53(23):6767-6775.