The family of graphene-based materials welcomed a new member, borophene, in 2014. Research on synthesis routes and experimental study on physicochemical and biological (especially in vivo) properties still is strongly desired in order to evaluate its practical potential as a drug delivery-system. The effect of two-dimensional borophene nanoflakes on cells, systems and the entire animal organism has not been studied so far. Therefore, we investigated in vivo its biocompatibility with hemocytes in the Tenebrio molitor as a model organism. Short-term studies demonstrated that borophene nanoflakes at doses of 0.5, 1 or 2 µg of nanoflakes per insect did not induce hemocytotoxicity. Hemocytes exposed to nanoflakes showed morphology, adhesiveness and ability to form filopodia as in the control hemocytes. A detailed study indicates that borophene nanoflakes do not: (i) generate intracellular reactive oxygen species in hemocytes, (ii) affect the mitochondrial membrane potential and (iii) interfere with phagocytosis. Therefore, this contribution presents new in vivo insights into the group of two-dimensional materials which are one of the most promising materials for biomedical applications owing to their special structure and unique properties. However, long-term studies in insects and other animals are still necessary to confirm that borophene is biocompatible and biologically safe.

To induce the water solubility of hexagonal boron nitride (h-BN), we exfoliated and functionalized bulk h-BN with hydroxyl groups (h-BN-OH-n). Short-term studies showed that h-BN-OH-n induced low cytotoxicity in different models: insect haemocytes (in vivo), human erythrocytes and mouse fibroblasts (in vitro). We also demonstrated that Alexa Fluor 647-h-BN-OH-n administered topically to the insects passed through the cuticle barrier and was phagocytosed by haemocytes. Nanoflakes did not affect the haemocyte cell membrane and did not interfere with the phagocytosis of latex beads. Long-term immunoassays showed that h-BN-OH-n, despite not inducing haemocytotoxicity, impaired nodulation, the most important cellular immune response in insects. The haemocytes exposed to h-BN-OH-n and then to bacteria differed in morphology and adhesiveness from the haemocytes exposed only to bacteria and exhibited the same morphology and adhesiveness as the control haemocytes. The h-BN-OH-n-induced decrease in nodulation can therefore result from the reduced ability of haemocytes to recognize bacteria, migrate to them or form microaggregates around them, which can lead to dysfunction of the immune system during pathogen infection. Long-term in vivo studies with animal models are still necessary to unambiguously confirm that h-BN is biocompatible and useful for application as a platform for drug delivery or for bioimaging.

At birth, the newborn fat-tailed dunnart relies on cutaneous gas exchange to meet metabolic demands, with continuous lung ventilation emerging several days later. We hypothesised that the delayed expression of lung ventilation (V_E) in these animals is in part due to a low responsiveness of the respiratory control system to blood gas perturbations. To address this hypothesis, we assessed the ventilatory and metabolic response to hypoxia (10% O₂) and hypercapnia (5% CO₂) using closed-system respirometry from birth to 23 days postpartum (P). Neonatal fat-tailed dunnarts displayed no significant hypoxic or hypercapnic ventilatory responses at any age. Regardless, significant hyperventilation through a suppression of metabolic rate (VO₂) was observed at birth in response to hypercapnia and in response to hypoxia at all ages, except P12. Therefore, reliance on cutaneous gas exchange during early life may be partially attributed to reduced chemosensitivity or a lack of central integration of chemosensitive afferent information. This may be in part due to the relative immaturity of this species at birth, compared with other mammals.