BALB/3T3 clone A31

Graphene Monolayer's Impact on Focal Cell Morphology and Size in BALB/3T3 Fibroblasts

Lasocka et al. investigated the effect of graphene scaffold on morphology and size in BALB/3T3 clone A31 fibroblasts and provides new data on biocompatibility of the "graphene-family nanomaterials".

The 24-hour cytotoxicity test with BALB/3T3 mouse fibroblasts showed comparable cell attachment and growth on graphene scaffolds and control substrates. Within the first 3 hours, cells adhered and flattened on both graphene and glass. Morphological changes began at 6 hours (Fig. 1), with cells transitioning to spindle shapes and extending pseudopods. By 12 hours, most cells became spindle-shaped with multiple pseudopods (Fig. 1), exhibiting intact nuclei and nucleoli. After 24 hours, pseudopods merged into thin lamellae. Cells grew normally on graphene without damage, detachment, or morphological differences versus controls (Fig. 1). BALB/3T3 cells exhibit contact inhibition, also known as contact paralysis, where cells avoid overlapping upon contact and change direction. Therefore, culturing them at the correct density (3 × 10⁴ cells/cm²) is crucial for proper morphology and size. After 12 hours, cells on graphene and control substrates measured 1638.6 ± 709.5 µm² and 1732.8 ± 711.2 µm², respectively (Fig. 2). Over the next 12 hours, their area increased by nearly half (Fig. 2), with no significant difference between graphene and glass.

Evaluation of Cytotoxicity of Hyaluronic Acid/Chitosan/Bacterial Cellulose-Based Membrane

Novel wound dressings must exhibit non-cytotoxic properties with cell viability exceeding 92%. In this study, Dechojarassri et al. compared the cytotoxicity of hyaluronic acid/chitosan/bacterial cellulose (BC(CS/HA)) membranes with alginate/chitosan/bacterial cellulose (BC(CS/Alg)) membranes.

BALB-3T3 clone A31 fibroblasts were initially grown at 1×10⁴ cells/well in a T75 flask for seven days before transferring them to 96-well plates for further incubation at 1, 3, and 5 days. Figure 3 is a fluorescence microscopy image of the live cells, while figure 4 shows a graph with the viability data. The blank (0% no membrane) and BC control showed 100% viability throughout the entire experiment. The BC(CS/Alg)5 and BC(CS/Alg)10 had decreased viability (around 82% at 5 days), while BC(CS/HA)5 and BC(CS/HA)10 had higher viability (94 and 87%, respectively). It was reported that the reason BC(CS/HA) has a greater biocompatibility was due to the HA being able to overcome CS caused cell damage. This was in-line with previously published data for the same CS/HA-based materials, which had 93-94% viability. In terms of wound dressing development, it was noted that BC(CS/HA)5 had 94% viability after 5 days, which is above the safety limit of 92% for wound dressings. Thus, the BC(CS/HA)5 membrane is a possible future development with antibiotics to improve wound healing.