Evaluating the Biocompatibility of Engineered Implants Using Nano-CaO2: A Cell Culture Study

This study aimed to evaluate the biocompatibility of engineered implants treated with varying concentrations of nano-CaO2 using RAW264.7 and BMSCs cell lines.

Cell Culture and Treatment:

RAW264.7 and BMSCs cells were utilized to assess the biocompatibility of engineered implants with different concentrations of nano-CaO2. Cells were seeded at a density of 5×10⁴ cells per well in a 24-well plate containing various samples. After co-culturing for 1 and 3 days, cells were incubated with 10% CCK-8 reagent for 3 hours to determine cell viability.

Cell Viability Assay:

Cell viability was semi-quantitatively assessed by measuring the absorbance at 450 nm using a microplate reader (BioTek Synergy Neo2, USA).

Cytoskeleton Staining and Morphological Analysis:

Morphological changes of the treated cells were evaluated using cytoskeleton staining. Cells were fixed with 4% formaldehyde, permeabilized with 0.5% Triton X-100, and subsequently stained for F-actin using 5% TRITC-phalloidin for 30 minutes in the dark. The nuclei were subsequently stained with DAPI (1:1000 v/v) for 10 minutes, and the cells were rinsed twice with PBS before examination using laser scanning confocal microscopy (LSCM).

Scanning Electron Microscopy (SEM) Preparation:

For SEM observation, cells were gently washed twice with sterile PBS and then immersed in 2.5% glutaraldehyde. After overnight fixation at 4°C, cells were rinsed with PBS and sequentially dehydrated in a series of ethanol-water mixtures (30%, 50%, 70%, 80%, 90%, 95%, and 100%) for 10 minutes each. Subsequently, critical point drying and gold sputtering were conducted prior to SEM examination.

Data Analysis:

Statistical analysis was performed using [State the statistical software and tests used]. Data are presented as mean ± standard deviation (SD). p-values less than 0.05 were considered statistically significant.

Results:

[Describe the findings of your study, including cell viability, morphological changes, and SEM observations.]

Discussion:

[Discuss the implications of your findings in the context of biocompatibility and the use of nano-CaO2 in engineered implants.]

Evaluating the Biocompatibility of Engineered Implants Using Nano-CaO2: A Cell Culture Study