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The acetone washes were repeated twice to remove the excessive cisplatin absorbing on the surface of mesospheres. After wash, the cisplatin-loaded mesospheres were air-dried at room temperature and stored at 4��C. Measurement of loading efficiency and cisplatin release To measure the loading efficiency of cisplatin in the mesospheres, about 1 to 2 mg of mesospheres were placed in a 15 ml centrifuge tube and then digested with at least 5 ml digest solution containing papain, protease K, ethylenediaminetetraacetic acid (EDTA) and cysteine. The suspension solutions containing mesospheres and enzymes were placed on a rotator in an incubator at 37��C for days. After digestion, the suspension solutions were centrifuged [http://www.selleckchem.com/products/abt-199.html this website] to precipitate the debris of MS and the supernatants were collected for analysis. To determine the cisplatin release from the mesospheres, the mesospheres loaded with CDDP/DMSO and CDDP/PBS for a minimal loading time ([http://www.selleckchem.com/products/pci-32765.html click here] solutions were prepared by diluting the platinum standard for ICP (TraceCERT?, 1000 mg/L Pt in hydrochloric acid, Fluka). Cytotoxicity of cisplatin released from cisplatin-loaded mesospheres A549 cells (ATCC #CCL-185; human [http://en.wikipedia.org/wiki/MMP23B MMP23B] lung adenocarcinoma epithelial cell line), NCI-H23 (ATCC #CRL-5800; human lung adenocarcinoma epithelial cell line), CRL-2081 (malignant mesothelioma cell line), and Lewis lung carcinoma cells were obtained from American Type Cell Collection (Manassas, VA). The cells were cultured in RPMI-1640 medium (Sigma, St Louis, MO) containing 10% fetal bovine serum (FBS), penicillin (100 U/ml), streptomycin (100 ?g/ml), fungizone (100 ?g/ml), 0.25% D-glucose, 0.2% sodium bicarbonate and 1% sodium pyruvate as reported earlier [49,50]. To determine the toxicity of cisplatin that was released out from the cisplatin-loaded mesospheres, the cisplatin released in PBS was collected first and reconstitute to desired concentration. About 10 mg of cisplatin-loaded mesospheres were dispersed in 2 ml PBS at room temperature to allow the release of cisplatin. After days of incubation, the dispersion solutions were centrifuged and the supernatant solution were collected and stored at room temperature. The concentration of the supernatant solution were determined by using ICP-AES and then diluted to 10 ?g/ml using PBS.