Journal: Pharmaceutics

Article Title: pH-Responsive Nanostructured Calcium Phosphate Microrods as Pulmonary Delivery Platform: Fabrication, Characterization, and Comparative Assessment of Cytotoxic and Transcriptomic Responses in Alveolar Macrophages

doi: 10.3390/pharmaceutics18040428

Figure Lengend Snippet: Internalization and interaction of MH-S cells after 3 h and 6 h of incubation time. MH-S cells were incubated with rhodamine-labeled CaP-Alg-Prot/CMC (3DL) for 3 h (top row) and 6 h (bottom row) and imaged by CLSM. Shown are single optical sections (DAPI, blue), F-actin (Phalloidin AF488, green) and microrods (rhodamine, red), as well as merged images. At both timepoints, microrods are successfully internalized by the cells. Image-based evaluation of 8 CLSM images per timepoint revealed that 37.8 ± 7.2% of cells after 3 h and 46.2 ± 14.6% after 6 h were classified as particle-positive (containing at least one internalized microrod or microrod-derived fragment). Boxed sections show different z-planes confirming the localization of the microrods beneath the F-actin-rich cortical region, rather than attached to the cell surface. 3D CLSM reconstructions of the images are shown in .

Article Snippet: SV-40-transformed murine alveolar macrophages (MH-S) (Cat. No. 300487) were purchased from Cytion (Eppelheim, Germany) and cultured in RPMI-1640 supplemented with 10% FCS in a humidified atmosphere of 5% carbon dioxide at 37 °C.

Techniques: Incubation, Labeling, Derivative Assay

Journal: Pharmaceutics

Article Title: pH-Responsive Nanostructured Calcium Phosphate Microrods as Pulmonary Delivery Platform: Fabrication, Characterization, and Comparative Assessment of Cytotoxic and Transcriptomic Responses in Alveolar Macrophages

doi: 10.3390/pharmaceutics18040428

Figure Lengend Snippet: Representative SEM ( a – c ) and CLSM ( d – f ) images obtained after 3 h of incubation of MH-S alveolar macrophages with CaP microrods, illustrating progressive stages of cell–particle interaction. In CLSM images, nuclei are shown in blue (DAPI), F-actin in green (Phalloidin), and microrods in red (rhodamine). ( a , d ) Early contact and particle probing mediated by membrane protrusions; insets provide a magnified view of protrusion–particle contact. ( b , e ) Uptake following alignment of the microrods along their shorter axis, consistent with a “tip-first” internalization process; insets highlight F-actin enrichment at rod–cell interface, indicative of phagocytic cup formation. ( c , f ) Advanced internalization of microrods.

Article Snippet: SV-40-transformed murine alveolar macrophages (MH-S) (Cat. No. 300487) were purchased from Cytion (Eppelheim, Germany) and cultured in RPMI-1640 supplemented with 10% FCS in a humidified atmosphere of 5% carbon dioxide at 37 °C.

Techniques: Incubation, Membrane

Journal: Pharmaceutics

Article Title: pH-Responsive Nanostructured Calcium Phosphate Microrods as Pulmonary Delivery Platform: Fabrication, Characterization, and Comparative Assessment of Cytotoxic and Transcriptomic Responses in Alveolar Macrophages

doi: 10.3390/pharmaceutics18040428

Figure Lengend Snippet: Cytotoxicity and cell viability of MH-S alveolar macrophages after 24 h of exposure time to CaP and SiO 2 microrods. ( a ) Cell viability assessed by MTT assay at different concentrations [mg/mL]. ( b ) Cell viability expressed as function of calculated rod/cell ratio. ( c ) Cytotoxicity assessed by LDH assay at different concentrations [mg/mL]. ( d ) Cytotoxicity expressed as function of calculated rod/cell ratio. CaP microrods consistently exhibited higher viability and lower cytotoxicity values than SiO 2 microrods. Each experiment was performed with n = 3–6 technical replicates, and data are presented as mean ± SD from N ≥ 3 independent experiments. The dotted lines indicate 80% viability and 20% cytotoxicity thresholds for visual reference. ( p < 0.01 **, p < 0.001 ***, p < 0.0001 ****) For full statistical analysis refer to .

Article Snippet: SV-40-transformed murine alveolar macrophages (MH-S) (Cat. No. 300487) were purchased from Cytion (Eppelheim, Germany) and cultured in RPMI-1640 supplemented with 10% FCS in a humidified atmosphere of 5% carbon dioxide at 37 °C.

Techniques: MTT Assay, Lactate Dehydrogenase Assay