Toxicity study of pro-angiogenic casein manganese oxide nanoparticles: an in vitro and in vivo approach

Abstract

Recently, we have demonstrated casein manganese oxide nanoparticles (CMnNP) that exhibit pro-angiogenic property established through different in vitro and in vivo experiments. The CMnNP was explored for therapeutic angiogenesis for treatment of wounds and recovery of hindlimb ischemia in pre-clinical mouse prototypical. It is well known that to translate any therapeutic nanoparticle for future clinical applications, their biosafety evaluation in small and large animals is essential. Herein, in the current study, the biosafety and bioavailability of the CMnNP have been explored by a systematic toxicity profiling study in C57BL/6J mice model. Initially, the in vitro cytotoxic effects of CMnNP were validated in RAW 264.7 cells. Later, the CMnNP was administered intraperitoneally with different doses (50, 300, and 2000 mg/kg b.wt./day) at different time points of exposure (acute: 2 weeks, sub-chronic: 4 weeks as well as chronic exposure: 8 and 20 weeks) with reference to the maximum tolerable dose (MTD) of CMnNP as per the OECD guidelines. The blood hematological and serum biochemical parameters of CMnNP treatment groups indicate negligible changes similar to untreated group. The histopathological examination of CMnNP-treated vital organs (lung, spleen, liver, brain, kidney, and heart) illustrates no major changes even at higher doses. Further, the biodistribution and excretion study depicts normal clearance of CMnNP. Additionally, the serum cytokine levels were normal in the therapeutic dose of CMnNP. The results altogether indicate that the non-toxic nature of CMnNP makes them useful as future therapeutic angiogenic agent for the treatment of various diseases where angiogenesis plays an important role.

Graphical Abstract

Keywords:

• Casein manganese oxide nanoparticles
• reactive oxygen species
• pre-clinical mouse model
• acute toxicity
• biodistribution

Acknowledgements

This research is supported by DST Nano Mission-project (SR/NM/NS-1252/2013; GAP 570), CRG-SERB-DST New Delhi (CRG/2022/004594, GAP0985), and PAN CSIR project (HCP0040) to CRP. SH and ST are thankful to CSIR and UGC, respectively, for their Research Fellowship. We thank Director, CSIR-IICT (Ms. No. IICT/Pubs./2023/314; dated 25 August 2023) for providing all the required facilities to carry out the work.

Author contributions

Shagufta Haque: planning, synthesis, in vitro assays, and in vivo animal experiments, data analysis, manuscript preparation. Sanchita Tripathy: contributed in animal experiments and writing of the manuscript. Yogesh Chandra: histopathological analysis and edited revised manuscript. Kathirvel Muralidharan: contributed in animal experiments. Chitta Ranjan Patra: conceived the original idea, design, overall planning, co-ordination guidance, and editing for final draft of the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

Data will be made available upon request from the authors.

Additional information

Funding

This research is supported by DST Nano Mission-project (SR/NM/NS-1252/2013; GAP 570), CRG-SERB-DST New Delhi (CRG/2022/004594, GAP0985), and PAN CSIR project (HCP0040) to CRP. SH and ST are thankful to CSIR and UGC, respectively, for their Research Fellowship.