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Molybdenum derived from nanomaterials incorporates into molybdenum enzymes and affects their activities in vivo

Abstract

Many nanoscale biomaterials fail to reach the clinical trial stage due to a poor understanding of the fundamental principles of their in vivo behaviour. Here we describe the transport, transformation and bioavailability of MoS2 nanomaterials through a combination of in vivo experiments and molecular dynamics simulations. We show that after intravenous injection molybdenum is significantly enriched in liver sinusoid and splenic red pulp. This biodistribution is mediated by protein coronas that spontaneously form in the blood, principally with apolipoprotein E. The biotransformation of MoS2 leads to incorporation of molybdenum into molybdenum enzymes, which increases their specific activities in the liver, affecting its metabolism. Our findings reveal that nanomaterials undergo a protein corona-bridged transport–transformation–bioavailability chain in vivo, and suggest that nanomaterials consisting of essential trace elements may be converted into active biological molecules that organisms can exploit. Our results also indicate that the long-term biotransformation of nanomaterials may have an impact on liver metabolism.

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Fig. 1: Partition coefficient of MoS2@HSA nanocomplexes in blood components: MoS2@HSA nanocomplexes rapidly interact with blood proteins and immune cells.
Fig. 2: Proteomic analysis of the protein corona and molecular dynamics simulation of the coronal blood proteins interacting with MoS2 2D nanodots.
Fig. 3: The biodistribution of MoS2@HSA nanocomplexes.
Fig. 4: The resident macrophages in the liver sinusoid and splenic red pulp sequester the MoS2@HSA nanocomplexes.
Fig. 5: The biotransformation and bioavailability of MoS2 as molybdenum cofactor in the liver.
Fig. 6: The fundamental interactions of nanoparticles with biological systems (nanoparticle–protein, nanoparticle–blood, nanoparticle–liver and nanoparticle–spleen) via a protein corona-bridged transport–transformation–bioavailability chain.

Data availability

The data supporting the findings of this study are available from the corresponding author upon reasonable request. The protein identification was conducted using UniProtKB Mus musculus sequence database (Swiss-Prot (560,459), UniProt release 2019_06, https://www.uniprot.org/). The protein classification and gene ontology were identified by Gene Ontology (molecular function and biological process) (http://geneontology.org/, 7 January 2019 release). Source data are provided with this paper.

Code availability

The original analysis code for molecular dynamics simulation is kept at the CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety at the Institute of High Energy Physics and is available from the corresponding authors upon reasonable request.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2016YFA0201600, 2020YFA0710700 and 2016YFE0133100), the Innovative Research Groups of the National Natural Science Foundation of China (11621505), the National Natural Science Foundation of China (31971311, 2202780088), the Program for International S&T Cooperation Projects of the Ministry of Science and Technology of China (2018YFE0117200), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB36000000), the CAS Interdisciplinary Innovation Team, the Bureau of International Cooperation Chinese Academy of Sciences (GJHG1852), the Research and Development Project in Key Areas of Guangdong Province (2019B090917011) and the Users with Excellence Project of Hefei Science Center CAS (2018HSC-UE004). We greatly appreciate the support from beamlines BL-15U1A and BL-14W1 (SSRF), beamlines 1W1B and 4B7A (BSRF) and beamline BL07W at the Hefei Light Source of the National Synchrotron Radiation Laboratory.

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Authors

Contributions

C.C. conceived the project and supervised the study. C.C., M.C. and R.C. designed the experiments. M.C. conducted the experiments with assistance from M.G., L.W., C.X., Y.W., Y.L. and S.X. L.Z., H.Y. and X.W. performed molecular dynamics simulation. Intravital confocal fluorescence microscopy imaging was conducted with help from Y.W. L.Z, L.W. and Y.W. assisted with collecting data of synchrotron radiation microbeam X-ray fluorescence imaging and X-ray absorption near-edge spectra. Y.G., Y.C., Y.W. and X.T. helped to collect and analyse the X-ray nano-CT imaging results. M.C., R.C., L.Z. and C.C. analysed results and wrote the manuscript. C.C. and Y.Z. edited the manuscript. All authors approved the manuscript.

Corresponding author

Correspondence to Chunying Chen.

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The authors declare no competing interests.

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Peer review information Nature Nanotechnology thanks the anonymous reviewers for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 Proteomic analysis of the protein corona (PC) and gene ontology classification.

a, Other components detected in protein corona. b, Relative abundance of proteins recognized by mononuclear phagocytic cells in the protein corona. Gene ontology analysis of the protein corona, molecular function (c) and biological process (d).

Source data

Extended Data Fig. 2 The molecular dynamics (MD) simulations to analyse the interactions between the MoS2 nanodots and fibrinogen (Fg), albumin (HSA) and apolipoprotein E (ApoE).

Root mean square deviation (RMSD, a), Salt-bridge (b) and H-bond (c) analysis in MD simulation of the interactions between 2H, 1 T MoS2 and fibrinogen (Fg), albumin (HSA) and apolipoprotein E (ApoE).

Source data

Extended Data Fig. 3 The biodistribution of MoS2@HSA nanocomplexes.

a, SRXRF imaging of Mo, S, Fe, K, Ca in liver at dose of 5.0 mg/kg (in terms of MoS2) at 6 h, 3 d, 14 d post-injection. SRXRF imaging of K, Ca in liver (b) and spleen (c) at dose of 5.0 mg/kg (in terms of MoS2) at 1 d, 7 d, 60 d post-injection. SRXRF images are representative of three independent experiments. d, The amounts of Mo in skin, muscle, bone, intestine, stomach and brain at the indicated time points after intravenous injection at the dose of 5.0 mg/kg. The data are presented as the mean ± S.D. n = 3 biologically independent mice. Scale bar: 1 mm.

Source data

Extended Data Fig. 4 Degradation and bioavailability of MoS2@HSA-derived Mo in the liver after treatment with dose of 0.25 mg/kg (MoS2 concentration).

a, Mo K-edge XANES (solid lines) and fitted curves (dashed lines) of liver preparations from mice treated with MoS2@HSA (0.25 mg MoS2/kg). b, Increased activity of molybdenum enzymes (AOX: Aldehyde oxidase; XOR: Xanthine oxidoreductase) in the liver of mice intravenously injected with MoS2 nanocomplex at a dose of 0.25 mg MoS2/kg (n = 3 biologically independent mice). c, Chemical forms and ratio of Mo in liver of mice treated with 0.25 mg MoS2/kg body weight fitted from the XANES spectra. n = 3 biologically independent experiments. The data are shown as the mean ± S.D. Statistical significance was tested with a two-tailed, unpaired Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001.

Source data

Extended Data Fig. 5 Degradability of MoS2@HSA in the spleen after treatment with dose of 5.0 mg/kg (MoS2 concentration).

a, Mo K-edge XANES (solid lines) and fitted curves (dash lines) of spleens from mice treated at the indicated times with MoS2 nanocomplex. b, Mo chemical forms and ratios in the samples were calculated based on the Mo K-edge XANES. The Data are shown as the mean ± S.D. c, Chemical forms and ratios of Mo in spleen of mice treated with 5.0 mg MoS2/kg body weight were fitted from the XANES spectra. n = 3 biologically independent experiments. Data are shown as mean ± S.D.

Source data

Extended Data Fig. 6 HSA and blood proteins retard the oxidation of MoS2 nanodots.

a, Mo K-edge XANES of bare MoS2, MoS2 coated with HSA and mouse blood proteins before and after incubation in PBS for 30 d at 37 °C with the presence of air. b, Mo chemical forms and ratios in the indicated samples were calculated based on the measurements of Mo K-edge XANES. Mo L3-edge and S K-edge XANES of reference materials with different valence states of Mo (c) and S (d), bare MoS2 and MoS2@HSA incubated in PBS at 37 °C with the presence of air for 6 h (e, f). 2D MoS2 nanodots incubated with H2O2 were set as positive control.

Source data

Supplementary information

Supplementary Information

Supplementary Figs. 1–11, Table 1, methods and refs. 1–12.

Reporting Summary

Supplementary Video 1

Intravital microscopy imaging of blood vessel.

Supplementary Video 2-1

3D nano-CT imaging of MoS2-treated neutrophil.

Supplementary Video 2-2

3D nano-CT imaging of MoS2-treated PBMCs.

Supplementary Video 2-3

3D nano-CT imaging of MoS2-treated platelets.

Supplementary Video 2-4

3D nano-CT imaging of control neutrophils.

Supplementary Video 2-5

3D nano-CT imaging of control PBMCs.

Supplementary Video 2-6

3D nano-CT imaging of control platelets.

Supplementary Video 3-1

Molecular dynamics trajectory of apolipoprotein E with 1T MoS2 nanodots.

Supplementary Video 3-2

Molecular dynamics trajectory of apolipoprotein E with 2H MoS2 nanodots.

Supplementary Video 3-3

Molecular dynamics trajectory of fibrinogen with 1T MoS2 nanodots.

Supplementary Video 3-4

Molecular dynamics trajectory of fibrinogen with 2H MoS2 nanodots.

Supplementary Video 3-5

Molecular dynamics trajectory of albumin with 1T MoS2 nanodots.

Supplementary Video 3-6

Molecular dynamics trajectory of albumin with 2H MoS2 nanodots.

Source data

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Cao, M., Cai, R., Zhao, L. et al. Molybdenum derived from nanomaterials incorporates into molybdenum enzymes and affects their activities in vivo. Nat. Nanotechnol. 16, 708–716 (2021). https://doi.org/10.1038/s41565-021-00856-w

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