viability assay kit  (Biotium)

Journal: Bioactive Materials

Article Title: Smart microenvironment-adaptive nanocatalytic hydrogel for sequential antibacterial, anti-inflammatory, and regenerative therapy of biofilm-infected wounds

doi: 10.1016/j.bioactmat.2026.02.043

Figure Lengend Snippet: Characterization, and Cytocompatibility Validation of HCOC. (A) Schematic illustration of the development of HCOC. (B) FTIR spectrum of OSA, CMCS and OC hydrogel. (C) Time-dependent evolution of gelation of OC and HCOC. (D) SEM images of HCOC and EDS mapping images of C, N, O and Cu for HCOC. (E) FTIR spectra of HC, OC and HCOC. (F) Dynamic frequency sweep measurements of OC and HCOC. (G) Frequency-dependent viscoelastic behavior of OC and HCOC. (H) Alternating strain sweep with alternating strains of 1% and 1000% at 100s intervals and (I) Self-healing behavior of HCOC. (J) Live/dead staining showing the metabolic activity of L929 and RAW 264.7 cells after treatment with HCOC for 48 h. Rates of proliferation of (K) L929 cells and (L) RAW 264.7 cells after treatment with PBS or HCOC. (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001).

Article Snippet: Following the protocol of the DMAO/PI Bacterial Live/Dead Staining Kit (Beyotime Biotechnology), the bacteria were incubated with a working solution containing both DMAO and PI dyes in the dark at room temperature for 15-20 min. Fluorescence microscopy imaging was subsequently carried out.

Techniques: Biomarker Discovery, Staining, Activity Assay

Journal: Bioactive Materials

Article Title: Smart microenvironment-adaptive nanocatalytic hydrogel for sequential antibacterial, anti-inflammatory, and regenerative therapy of biofilm-infected wounds

doi: 10.1016/j.bioactmat.2026.02.043

Figure Lengend Snippet: pH Self-Adaptive Antioxidant Capacity of HCOC (Stage II: anti-inflammation). Cu ion release behavior of (A) HC (1 mg/mL) and (B) HCOC (1 mg/mL) at different pH levels. (C) ABTS + and (D) H 2 O 2 scavenging activity at different pH of Cu 5.4 O, HC and HCOC. (E) O 2 ∙ - , (F)∙OH scavenging activity of Cu 5.4 O, HAs, HC, HCOC. (G) SOD-like, (H) CAT-like and (I) GPx-like activities of HCOC. (J) Fluorescence images showing intracellular ROS detection by DCFH-DA staining, live/dead staining images and (K) cell viability of L929 cells with different treatments (All groups received 500 μM H 2 O 2 and different HCOC concentrations (I: PBS; II: 0; III: 0.25; IV: 0.50; V: 1.0 mg/mL HCOC). (L) Quantitative analysis of the cells under different treatments. (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001) (M) Schematic illustration of pH-responsive activity and ROS scavenging and alleviating cellular oxidative stress of HCOC.

Article Snippet: Following the protocol of the DMAO/PI Bacterial Live/Dead Staining Kit (Beyotime Biotechnology), the bacteria were incubated with a working solution containing both DMAO and PI dyes in the dark at room temperature for 15-20 min. Fluorescence microscopy imaging was subsequently carried out.

Techniques: Activity Assay, Fluorescence, Staining

Journal: Bioactive Materials

Article Title: Near-infrared light-driven photocatalytic reactive oxygen species-generating antibacterial and self-shrinking hybrid hydrogels for combating drug-resistant bacterial biofilm infection and accelerating wound healing

doi: 10.1016/j.bioactmat.2025.12.049

Figure Lengend Snippet: Cytotoxicity and cytology evaluations of the Co-BOS@C/F Gel. Cell viability with different hydrogel treatment in (a) L929 cells and (b) HUVECs at different concentrations for 24 h and 48 h; Fluorescence images of live/dead cells of HUVECs using a Calcein-AM/PI assay. The cells were co-cultured with the media extracts of different hydrogels, incubated for (c) 1 D and (d) 3 D. (e) Scratch assay for HUVECs treated with C/F Gel, BOS@C/F Gel, and Co-BOS@C/F Gel. Data are presented as mean value ± SD. ( ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; NS, not significant).

Article Snippet: Dulbecco's modified Eagle's medium (DMEM) medium and Roswell Park Memorial Institute (RPMI) 1640 medium containing 1 % penicillin/streptomycin were obtained from Jiangsu KeyGEN Biotech Co., Ltd. DNA Damage Assay Kit by γ-H2AX Immunofluorescence, Beyo3DTM ROS Assay Kit with DCFH-DA (S1105S), Live/Dead Bacterial Staining Kit with DMAO & PI was obtained to Beyotime Biotechnology (Shanghai) Co., Ltd. Double-mixed injector (suit), MixDS was obtained to Yongqinquan (Suzhou) Intelligent Equipment Co., Ltd. FBS was obtained to Inner Mongolia Jinyuankang Biotechnology Co., Ltd.

Techniques: Fluorescence, Cell Culture, Incubation, Wound Healing Assay