Survival, growth and digestive functions after exposure to nanodiamonds - Transgenerational effects beyond contact time in house cricket strains.

The long-term exposure effects of nanodiamonds (NDs), spanning an organism's entire lifespan and continuing for subsequent generation, remain understudied. Most research has focused on evaluating their biological impacts on cell lines and selected organisms, typically over short exposure durations lasting hours or days. The study aimed to assess growth, mortality, and digestive functions in wild (H) and long-lived (D) strains of Acheta domesticus (Insecta: Orthoptera) after two-generational exposure to NDs in concentrations of 0.2 or 2 mg kg-1 of food, followed by their elimination in the third generation. NDs induced subtle stimulating effect that depended on the strain and generation. In the first generation, more such responses occurred in the H than in the D strain. In the first generation of H strain insects, contact with NDs increased survival, stimulated the growth of young larvae, and the activity of most digestive enzymes in mature adults. The same doses and exposure time did not cause similar effects in the D strain. In the first generation of D strain insects, survival and growth were unaffected by NDs, whereas, in the second generation, significant stimulation of those parameters was visible. Selection towards longevity appears to support higher resistance of the insects to exposure to additional stressor, at least in the first generation. The cessation of ND exposure in the third generation caused potentially harmful changes, which included, e.g., decreased survival probability in H strain insects, slowed growth of both strains, as well as changes in heterochromatin density and distribution in nuclei of the gut cells in both strains. Such a reaction may suggest the involvement of epigenetic inheritance mechanisms, which may become inadequate after the stress factor is removed.

Quantitative Evaluation of the Cellular Uptake of Nanodiamonds by Monocytes and Macrophages.

Fluorescent nanodiamonds (FNDs) with negative nitrogen-vacancy (NV- ) defect centers are great probes for biosensing applications, with potential to act as biomarkers for cell differentiation. To explore this concept, uptake of FNDs (≈120 nm) by THP-1 monocytes and monocyte-derived M0-macrophages is studied. The time course analysis of FND uptake by monocytes confirms differing FND-cell interactions and a positive time-dependence. No effect on cell viability, proliferation, and differentiation potential into macrophages is observed, while cells saturated with FNDs, unload the FNDs completely by 25 cell divisions and subsequently take up a second dose effectively. FND uptake variations by THP-1 cells at early exposure-times indicate differing phagocytic capability. The cell fraction that exhibits relatively enhanced FND uptake is associated to a macrophage phenotype which derives from spontaneous monocyte differentiation. In accordance, chemical-differentiation of the THP-1 cells into M0-macrophages triggers increased and homogeneous FND uptake, depleting the fraction of cells that were non-responsive to FNDs. These observations imply that FND uptake allows for distinction between the two cell subtypes based on phagocytic capacity. Overall, FNDs demonstrate effective cell labeling of monocytes and macrophages, and are promising candidates for sensing biological processes that involve cell differentiation.

Phytotoxic effect and molecular mechanism induced by nanodiamonds towards aquatic Chlorella pyrenoidosa by integrating regular and transcriptomic analyses.

The growing diverse applications of nanodiamonds (NDs), especially as adsorbents and catalysts for wastewater treatment, have significantly increased their discharge and potential risk towards aquatic ecosystems. Although NDs have been certified for superior biocompatibility and lower toxicity towards numerous human cell lines, the characteristic response and underlying mechanism of aquatic microalgal response remains unclear. Here, the response of Chlorella pyrenoidosa to five concentrations of NDs was thoroughly investigated by comprehensive phenotypic and transcriptional examinations. Results indicated that higher concentration of NDs (50 mg/L) induced 75.4% growth inhibition, exacerbated oxidative stress and malformed morphology of microalgae after 48 h exposure. Meanwhile, the aggregated microalgae formed several flocs, apparently under 50 mg/L NDs. Noticeably, photosynthesis was susceptible to the NDs exposure. Although, the chlorophyll content and genes involved in photosynthesis were significantly improved by NDs, the results obtained from the photochemical parameters indicated that the excessive electrons during photosynthesis might be a pivotal reason for oxidative stress generation. Additionally, the genes included in amino acids metabolism and protein synthesis were up-regulated to alleviate the oxidative stress. Collectively, this work discloses the explicit molecular mechanisms of aquatic microalgae and provides comprehensive insights of potential aqueous environmental risk of gradually emergent NDs.

Coated nanodiamonds interact with tubulin beta-III negative cells of adult brain tissue.

Fluorescent nanodiamonds (NDs) coated with therapeutics and cell-targeting structures serve as effective tools for drug delivery. However, NDs circulating in blood can eventually interact with the blood-brain barrier, resulting in undesired pathology. Here, we aimed to detect interaction between NDs and adult brain tissue. First, we cultured neuronal tissue with ND ex vivo and studied cell prosperity, regeneration, cytokine secretion, and nanodiamond uptake. Then, we applied NDs systemically into C57BL/6 animals and assessed accumulation of nanodiamonds in brain tissue and cytokine response. We found that only non-neuronal cells internalized coated nanodiamonds and responded by excretion of interleukin-6 and interferon-γ. Cells of neuronal origin expressing tubulin beta-III did not internalize any NDs. Once we applied coated NDs intravenously, we found no presence of NDs in the adult cortex but observed transient release of interleukin-1α. We conclude that specialized adult neuronal cells do not internalize plain or coated NDs. However, coated nanodiamonds interact with non-neuronal cells present within the cortex tissue. Moreover, the coated NDs do not cross the blood-brain barrier but they interact with adjacent barrier cells and trigger a temporary cytokine response. This study represents the first report concerning interaction of NDs with adult brain tissue.

The sp3/sp2 carbon ratio as a modulator of in vivo and in vitro toxicity of the chemically purified detonation-synthesized nanodiamond via the reactive oxygen species generation.

Nanodiamonds have been suggested as biocompatible materials and are suitable for various biomedical applications, but little is known about how to synthesize safer nanodiamonds. Herein, seven different detonation-synthesized nanodiamonds (DNDs) with sequential sp3/sp2 carbon ratios were assembled by controlling the chemical purification parameters and the role of sp3/sp2 carbon ratio on the toxicity of DNDs was investigated. Carbon black and nickel oxide nanoparticles were used as reference particles. The intrinsic reactive oxygen species (ROS) generation potential of DNDs was estimated by a 2'7'-dichlorofluorescein diacetate (DCFH-DA) assay, and these values showed a good negative correlation with the sp3/sp2 carbon ratios, which implies that ROS generation increased as the sp3/sp2 carbon ratio decreased. As a model to investigate inflammogenic potential of DND samples, a rat intratracheal instillation model was used as the lung is very sensitive to nanoparticle exposures. The sp3/sp2 carbon ratios or the estimated values of ROS generation potential showed excellent linear correlations with the number of neutrophils and pro-inflammatory cytokines in bronchoalveolar lavage fluid at 24 h after instillation. Treatment of DND samples to THP-1 derived macrophages also showed that the sp3/sp2 carbon ratios or the estimated values of ROS generation potential were closely related with the toxicity endpoints such as cell viability and pro-inflammatory cytokines. Taken together, these data demonstrate that the sp3/sp2 carbon ratio is the key determinant for the toxicity of DNDs, which can be a useful tool for the safer-by-design approach of DNDs and the safety assessment of carbon nanoparticles.

Surface functionalization of nanodiamonds for biomedical applications.

Nanodiamonds (NDs), recent member of carbon nanomaterial, are nano-scale carbon allotropes having versatile surface chemistry. NDs are commonly synthesized by detonation and followed by purification, surface modification and surface functionalization. Surface functionalization of NDs enhances safety, bio-compatibility and lowers toxicity. It involves initial surface homogenization followed by attachment of ligand on NDs which increases hydrophobicity, reduces surface charge and improves surface chemistry. Generally, surface functionalization is carried out by covalent and non-covalent attachment and in biomedical applications various functional groups, biomolecules, or polymers can be attached to NDs. This review is focused on surface functionalization methods for NDs and their biomedical applications. Surface functionalization is beneficial to improve physicochemical properties of NDs which may be further utilized in diagnosis and targeted drug delivery.

The Effects of Paclitaxel in the Combination of Diamond Nanoparticles on the Structure of Human Serum Albumin (HSA) and Their Antiproliferative Role on MDA-MB-231cells.

Since the interactions of anti-cancer agents with blood constituents, in particular with human serum albumin (HSA) may have a major impact on drug pharmacology, the present study designed to provide a fundamental understanding of the interaction of nanodiamonds (NDs) together with paclitaxel (PTX) with HSA in detail for the first time. The UV-Vis, steady-state fluorescence, far-UV CD and zeta potential results displayed that PTX + NDs could form a complex with HSA. Additionally, the values of binding constants and ΔG° showed that PTX + NDs interact strongly with HSA compared to PTX or NDs alone and the hydrophobic force plays a major role in this interaction. Moreover, the in vitro release behavior of PTX + NDs form HSA can be regulated at dissimilar pH levels. The anticancer property of 0.5 µM PTX + 20 µM NDs by MTT assay demonstrates that this combination can tremendously diminish the proliferation of MDA-MB-231cells compared to PTX or NDs alone. Altogether, the structure of HSA changed moderately in the presence of PTX + NDs and PTX + NDs can promote mortality of MDA-MB-231 cells besides those mortality effects induced via PTX or NDs alone. The results obtained from this study can help in understanding the pharmacokinetic properties of PTX + NDs.

Toxicity of nanodiamonds to white rot fungi Phanerochaete chrysosporium through oxidative stress.

Nanodiamonds (NDs) are produced with large scale and applied in many areas, thus the environmental impacts and hazards of NDs should be systematically investigated. In this study, we evaluated the interaction between detonation NDs and white rot fungus Phanerochaete chrysosporium and the impact on the fungus decompositions activities. NDs did not influence the biomass gain of P. chrysosporium and the culture medium pH values. The mycelia of P. chrysosporium were destroyed upon the direct contact with NDs, while the rest retained the fibrous structure. Ultrastructural observations suggested that small aggregates of NDs seldom entered the fungus cells, but the break of cell wall and the loss of cytoplasm were induced by NDs. Under both optical and electron microscopes, the aggregation of colloidal ND particles was observed, which was the possible reason of low toxicity. High concentrations of NDs inhibited the laccase activity and manganese peroxidase activity of P. chrysosporium, which led to the decrease of decomposition activity for pollutants. Colloidal ND particles were not well dispersed in sawdust degradation evaluations, so no inhibitive effect was observed for wood degradation. The toxicological mechanism of NDs was assigned to oxidative stress. The results collectively suggested that NDs had low toxicity to white rot fungi and could be applied safely. The colloid dispersion/aggregation of nanoparticles in biological systems should be carefully considered during the design of safe nanomaterials.

Nanodiamonds of Different Surface Chemistry Influence the Toxicity and Differentiation of Rat Bone Mesenchymal Stem Cells In Vitro.

The nanodiamonds (NDs) have attracted much attention in biomedical applications due to their excellent magnetic and optical properties. However, comprehensive study of different surfacemodified NDs on toxicity and differentiation of mesenchymal stem cells are very deficient. In this study, three types of NDs, i.e., ND-COOH, ND-NH+3 and ND-PEG were co-cultured with rat bone mesenchymal stem cells (MSCs) to assess their biosafety and effects on differentiation. In a dry state, they had a similar diameter of about 6-7 nm, and aggregated into ˜100 nm (hydrodynamic size) in cell culture medium. Co-culture with MSCs showed that the ND-COOH and ND-PEG had lower cytotoxicity than ND-NH+3. Alkaline comet assay showed slight genotoxicity for all the NDs regardless of their surface coatings. The reactive oxygen species (ROS) test showed that the cytotoxicity and genotoxicity of NDs may be attributed to the NDs-mediated intracellular oxidative stress. All the NDs did not show significant impact on the osteogenic differentiation of MSCs, whereas the ND-COOH and ND-PEG slightly impaired the adipogenic differentiation. Taken together, these findings provide some momentous implications for the design of surface chemical structures of NDs for their applications in biological field.

Production, surface modification and biomedical applications of nanodiamonds: A sparkling tool for theranostics.

Diamond nano-particles or nanodiamonds (NDs) are nano-scale carbon allotropes and exhibit characteristic mechanical and optical properties. Unlike other nanocarbon materials NDs are highly bio-compatible and comparatively safe which makes them the material of future interest in the field of medicine. NDs are synthesized by various techniques which includes detonation technique, chemical vapour deposition, high pressure high temperature etc. and followed by post-synthesis processing such as purification, surface modification and functionalization. Interestingly, the versatile surface chemistry of NDs facilitates the conjugation with various functional groups and chemical moieties, which can be utilized for biomedical applications. Due to their unique optical and spectroscopic properties, NDs have been studied in bio-imaging in which NDs doped with specific atoms such as nitrogen, sodium, boron and were used for cell labelling and/or cell tracking. NDs have also been explored as a targeted drug delivery platform in conjugation with various therapeutic agents such as anti-cancer, anti-bacterial, anti-inflammatory agents etc., attributed to its flexible surface properties. Apart from this, macromolecules like protein and nucleic acids can be delivered in an efficient and safe manner with NDs. This review focuses on the synthesis, surface modification and various biomedical applications of nanodiamonds as a theranostic agent.

Functionalization of stable fluorescent nanodiamonds towards reliable detection of biomarkers for Alzheimer's disease.

BACKGROUND: Stable and non-toxic fluorescent markers are gaining attention in molecular diagnostics as powerful tools for enabling long and reliable biological studies. Such markers should not only have a long half-life under several assay conditions showing no photo bleaching or blinking but also, they must allow for their conjugation or functionalization as a crucial step for numerous applications such as cellular tracking, biomarker detection and drug delivery. RESULTS: We report the functionalization of stable fluorescent markers based on nanodiamonds (NDs) with a bifunctional peptide. This peptide is made of a cell penetrating peptide and a six amino acids long ß-sheet breaker peptide that is able to recognize amyloid ß (Aß) aggregates, a biomarker for the Alzheimer disease. Our results indicate that functionalized NDs (fNDs) are not cytotoxic and can be internalized by the cells. The fNDs allow ultrasensitive detection (at picomolar concentrations of NDs) of in vitro amyloid fibrils and amyloid aggregates in AD mice brains. CONCLUSIONS: The fluorescence of functionalized NDs is more stable than that of fluorescent markers commonly used to stain Aß aggregates such as Thioflavin T. These results pave the way for performing ultrasensitive and reliable detection of Aß aggregates involved in the pathogenesis of the Alzheimer disease.

Chronic toxicity of nanodiamonds can disturb development and reproduction of Acheta domesticus L.

The use of nanodiamonds in numerous materials designed for industry and medicine is growing rapidly. Consequently health and environmental risks associated with the exposure of humans and other biota to nanodiamonds-based materials are of the utmost importance. Scarcity of toxicological data for these particles led us to examine the potentially deleterious effects of nanodiamonds in model insect species, Acheta domesticus (Orthoptera) chronically exposed to ND in its diet. Organism-level end-point indices (lifespan, body weight, consumption, caloric value of faeces, reproduction) revealed adverse changes in the treated crickets in comparison with the control. Preliminary studies of oxidative stress level in the offspring of ND-treated crickets suggest toxicity of these particles limited to the exposed individuals. EPR analysis showing increase of radical signal in the faeces of ND-fed crickets led us to propose novel mechanism of nanodiamonds toxicity that is discussed in the light of literature data. CAPSULE: Development and reproduction of Acheta domesticus can be disturbed by the chronic exposure to nanodiamonds.

Fluorescent Nanodiamonds in Biomedical Applications.

Nanoparticles have an extended surface and a large surface area, which is the ratio of the size of the surface area to the volume. A functionalized surface can give rise to more modifications and therefore allows this nanomaterial to have new properties. Fluorescent molecules contain fluorophore, which is capable of being excited via the absorption of light energy at a specific wavelength and subsequently emitting radiation energy of a longer wavelength. A chemically modified surface of nanodiamond (ND; by carboxylation) demonstrated biocompatibility with DNA, cytochrome C, and antigens. In turn, fluorescent nanodiamonds (FNDs) belong to a group of new nanomaterials. Their surface can be modified by joining functional groups such as carboxyl, hydroxyl, or amino, after which they can be employed as a fluorescence agent. Their fluorescent properties result from defects in the crystal lattice. FNDs reach dimensions of 4-100 nm, have attributes such as photostability, long fluorescence lifetimes (10 ns), and fluorescence emission between 600 and 700 nm. They are also nontoxic, chemically inert, biocompatible, and environmentally harmless. The main purpose of this article was to present the medical applications of various types of modified NDs.

In vitro cytotoxicity assessment of nanodiamond particles and their osteogenic potential.

Scaffolds functionalized with nanodiamond particles (nDP) hold great promise with regard to bone tissue formation in animal models. Degradation of the scaffolds over time may leave nDP within the tissues, raising concerns about possible long-term unwanted effects. Human SaOS-2 osteoblast-like cells and U937 monoblastoid cells were exposed to five different concentrations (0.002-2 mg/L) of nDP (size range: 2.36-4.42 nm) for 24 h. Cell viability was assessed by impedance-based methods. The differential expression of stress and toxicity-related genes was evaluated by polymerase chain reaction (PCR) super-array, while the expression of selected inflammatory and cell death markers was determined by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). Furthermore, the expression of osteogenic genes by SaOS-2 cells, alkaline phosphatase activity and the extracellular calcium nodule deposition in response to nDP were determined in vitro. Cells responded differently to higher nDP concentrations (≥0.02 mg/L), that is, no loss of viability for SaOS-2 cells and significantly reduced viability for U937 cells. Gene expression showed significant upregulation of several cell death and inflammatory markers, among other toxicity reporter genes, indicating inflammatory and cytotoxic responses in U937 cells. Nanodiamond particles improved the osteogenicity of osteoblast-like cells with no evident cytotoxicity. However, concentration-dependent cytotoxic and inflammatory responses were seen in the U937 cells, negatively affecting osteogenicity in co-cultures. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1697-1707, 2018.

In Vitro Immune Activation of Raw264.7 Cells by Nanodiamonds with Different Surface Chemistry.

In this study, three commercially available nanodiamonds (NDs) of similar size (about 5 nm in diameter) but with different surface chemistry (-COOH, -NH2 and -poly(ethylene glycol)) were used to study the toxicity and immune activation of Raw264.7 cells. Only ND-PEG is well dispersed in water and cell culture medium (about 10 nm in diameter), while ND-COOH and ND-NH2 only showed some aggregation, about 50 nm in water and about 100 nm in cell culture medium. The three NDs showed different zeta potentials in water but the difference disappeared in cell culture medium due to the surface adsorption of the proteins. The ND-PEG did not show any obvious signs of cytotoxicity or activation on Raw264.7 cells under tested conditions. The ND-COOH and ND-NH2 caused dose-dependent toxicity, with the amino group capped NDs being much more toxic compare to those of the carboxylic acid group at the same exposure conditions. ND-COOH induced a certain immune response in Raw264.7 cells, leading to a higher expression of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6), especially with a significantly longer incubation time. ND-NH2 also induced significant expression of TNF-α and IL-6 in Raw264.7 cells at first, but this expression was reduced with a longer incubation time due to cell death.

Recombinant Protein Polymers for Colloidal Stabilization and Improvement of Cellular Uptake of Diamond Nanosensors.

Fluorescent nanodiamonds are gaining increasing attention as fluorescent labels in biology in view of the fact that they are essentially nontoxic, do not bleach, and can be used as nanoscale sensors for various physical and chemical properties. To fully realize the nanosensing potential of nanodiamonds in biological applications, two problems need to be addressed: their limited colloidal stability, especially in the presence of salts, and their limited ability to be taken up by cells. We show that the physical adsorption of a suitably designed recombinant polypeptide can address both the colloidal stability problem and the problem of the limited uptake of nanodiamonds by cells in a very straightforward way, while preserving both their spectroscopic properties and their excellent biocompatibility.

Carboxylated nanodiamonds can be used as negative reference in in vitro nanogenotoxicity studies.

Nanodiamonds (NDs) are promising nanomaterials for biomedical applications. However, a few studies highlighted an in vitro genotoxic activity for detonation NDs, which was not evidenced in one of our previous work quantifying γ-H2Ax after 20 and 100 nm high-pressure high-temperature ND exposures of several cell lines. To confirm these results, in the present work, we investigated the genotoxicity of the same 20 and 100 nm NDs and added intermediate-sized NDs of 50 nm. Conventional in vitro genotoxicity tests were used, i.e., the in vitro micronucleus and comet assays that are recommended by the French National Agency for Medicines and Health Products Safety for the toxicological evaluation of nanomedicines. In vitro micronucleus and in vitro comet assays (standard and hOGG1-modified) were therefore performed in two human cell lines, the bronchial epithelial 16HBE14o- cells and the colon carcinoma T84 cells. Our results did not show any genotoxic activity, whatever the test, the cell line or the size of carboxylated NDs. Even though these in vitro results should be confirmed in vivo, they reinforce the potential interest of carboxylated NDs for biomedical applications or even as a negative reference nanoparticle in nanotoxicology. Copyright © 2017 John Wiley & Sons, Ltd.

Phagocytosis and immune response studies of Macrophage-Nanodiamond Interactions in vitro and in vivo.

The applications of nanodiamond as drug delivery and bio-imaging can require the relinquishing ND-drug conjugate via blood flow, where interaction with immune cells may occur. In this work, we investigated the ND penetration in macrophage and the immune response using the tissue-resident murine macrophages (RAW 264.7). Confocal fluorescence imaging, immunofluorescence analysis of nuclear translocation of interferon regulatory factor IRF-3 and transcriptional factor NF-κΒ, analysis of pro-inflammatory cytokines production IL-1ß, IL-6 IL-10 with a reverse transcription-polymerase chain reaction technique were applied. The TNF-α factor production has been studied both in vitro at ND interaction with the macrophage and in vivo after ND injection in the mice blood system using immunoassay. The macrophage antibacterial function was estimated through E. coli bacterial colony formation. ND didn't stimulate the immune response and functionality of the macrophage was not altered. Using MTT test, ND was found negligibly cytotoxic to macrophages. Thus, ND can serve as a biocompatible platform for bio-medical applications. Left: Graphic representation of Nanodiamond internalization in macrophage. Right: (a) Fluorescence images of lysosomes, (b) nanodiamond and (c) merged image of nanodiamond internalization in macrophage.

[INFLUENCE OF NANODIAMONDS AND CARBON NANOWIRES ON SURVIVAL AND CELLS STRUCTURE IN CHICKEN EMBRYO].

Aim - to determine the effect of nanodiamonds and carbon nanowires on the survival and ultrastructure of chicken embryo cells. The experiment was carried out on chicken embryos, incubated from eggs of Hy-Line breed. Control and two experimental groups were formed (total number of embryos - 100). Diamond nanoparticles and carbon nanowires were administered on day 3 of incubation as a suspension of a biocompatible dextran. Ultrastructural analysis and general study of embryos state were carried out. The most expressed pathological effects were observed in the group with the introduction of the CNW, which caused visual impairment of embryogenesis that started from the early incubation periods. As for ND we can claim their prolonged impact on the development of embryos, manifested in the gradual deterioration of the embryos condition with the manifestations of the pathology in the provisory organs and the body of embryos. The results of our study demonstrate that both types of nanostructures can cause sublethal and irreversible morphologic changes. Detection of morphological evidence of the impact of nanomaterials at significant distances from the site of administration of nanoparticles shows highly penetrating ability of nanomaterials. The presence of damages specific for each type of nanoparticles shows affinity to various tissues and cellular structures. It is demonstrated that similar, at first glance, impact of nanomaterials, such as the induction of oxidative stress might be caused by specific structural transformations. So, ND cause vacuolization of mitochondria, and the CNW - deformation of their shape and appearance of dark inclusions in them.

Nanodiamonds for Medical Applications: Interaction with Blood in Vitro and in Vivo.

Nanodiamonds (ND) have emerged to be a widely-discussed nanomaterial for their applications in biological studies and for medical diagnostics and treatment. The potentials have been successfully demonstrated in cellular and tissue models in vitro. For medical applications, further in vivo studies on various applications become important. One of the most challenging possibilities of ND biomedical application is controllable drug delivery and tracing. That usually assumes ND interaction with the blood system. In this work, we study ND interaction with rat blood and analyze how the ND surface modification and coating can optimize the ND interaction with the blood. It was found that adsorption of a low concentration of ND does not affect the oxygenation state of red blood cells (RBC). The obtained in vivo results are compared to the results of in vitro studies of nanodiamond interaction with rat and human blood and blood components, such as red blood cells and blood plasma. An in vivo animal model shows ND injected in blood attach to the RBC membrane and circulate with blood for more than 30 min; and ND do not stimulate an immune response by measurement of proinflammatory cytokine TNF-α with ND injected into mice via the caudal vein. The results further confirm nanodiamonds' safety in organisms, as well as the possibility of their application without complicating the blood's physiological conditions.