Co-delivery of methotrexate and extracted berberine from berberine vulgaris-Lby PAMAM dendrimer for evaluation of their anticancer effect on HeLa cancer cell line compared to each one berberin and methotrexate effect alone invitro
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Background and objective: Cervical cancer is the second most common cancer after breast cancer among women throughout the world. Conventional strategies treatments include chemotherapy, surgery and radiotherapy. Chemotherapy is more selective to treat cervical cancer but unfavorable side effects limited it applications. Combination treatment and a variety of drug delivery systems including nanoparticles are used to reduce the complications of chemotherapy. Materials and Methods: The formation of MTX conjugated PAMAM transform loaded with BER (MTX-PAMAM-BER) had been evaluated by (FT-IR), and particle size evaluated through (DLS). In regarding to biological assay, MTT assay was carried out to estimate the cytotoxicity of one and combined groups, followed by apoptosis assay to calculate the number of apoptotic cells. Both DAPI staining and DNA ladder were complemented tests to illuminate morphological changes and DNA fracturing on HeLa cancer cells. Results: The characterization of formulated nanostructure showed that the conjugation of MTX and loading and BER into PAMAM dendrimer was carried of properly. The obtained results from MTT, flow cytometry, fluorescence microscopy and DNA fragmentation assays revealed that presence of MTX on PAMAM nanoparticles, particularly the co-delivey system (PAMAM-MTX-BER) had higher cell cytotoxicity, apoptosis and DNA fragmentation against HeLa cells over 48 hour. Conclusio: BER mixed with MTX can be considered as an effective therapy to reduce growth and induce apoptosis in HeLa cancer cells. Also, surface modification of PAMAM nanoparticle with MTX and BER make it a suitable platform for active delivery drugs into cancer cells and had higher drug transfection efficiency.