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Human intervertebral discs harbour a unique microbiome and dysbiosis determines health and disease

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Abstract

Background

To document the role of sub-clinical infections in disc disorders and investigate the existence of microbiome in intervertebral discs (IVD).

Methods

Genomic DNA from 24 lumbar IVDs [8—MRI normal discs (ND) from brain dead yet alive organ donors, 8—disc herniation (DH), 8—disc degeneration (DD)] was subjected to 16SrRNA sequencing for profiling the diversity of human disc microbiome in health and disease. The disc microbiome was further compared to established human gut and skin microbiomes.

Results

All healthy MRI normal discs from brain dead yet alive organ donors also had a rich bacterial presence. A total of 424 different species (355-ND, 346-DD, and 322-DH) were detected, with 42.75% OTUs being classified at kingdom level, 44% at the phylum level, 22.62% at genus level, and 5.5% at species level. Varying biodiversity and abundance between healthy and diseased discs were documented with protective bacteria being abundant in normal discs, and putative pathogens abundant in DD and DH. Propionibacterium acnes had a similar but lower abundance to other pathogens in all three groups ND (3.07%), DD (3.88%), DH (1.56%). Fifty-eight bacteria were common between gut and IVD microbiomes, 29 between skin and IVD microbiomes, and six common to gut/skin/IVD.

Conclusion

Our study challenges the hitherto concept of sterility in healthy IVD and documented a microbiome even in MRI normal healthy discs. The varying abundance of bacteria between ND, DD, and DH documents ‘dysbiosis’ as a possible etiology of DD. Many known pathogens were identified in greater abundance than Propionibacterium acnes, and there was evidence for the presence of the gut/skin/spine microbiome axis.

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Abbreviations

16S rRNA:

16 (Svedberg) ribosomal ribonucleic acid

30S:

Ribosomal subunit contains 16S rRNA

50S:

Ribosomal subunit

DADA2:

Data analysis decision and action

DD:

Disc degeneration

DH:

Disc herniation

DNA:

Deoxyribose nucleic acid

ESI:

Electrospray ionization

GGv138:

Green GENES database version 138

IRB:

Intuitional Review Board

LBP:

Low back pain

LC:

Liquid chromatography

Mascot:

Software search engine to identify proteins uses MOWSE algorithm

MRI:

Magnetic resonance imaging

MS/MS:

Tandem mass spectrometry

MW test:

Mann–Whitney test

ND:

Normal disc

OTU:

Operational taxonomic unit

PICRUSt:

Phylogenetic investigation of communities by reconstruction of unobserved states

q2:

QIIME2

QIIME:

Quantitative insights into microbial ecology

R packages:

Statistical tool designed by Ross Ihaka and Robert gentleman

Sequest HT:

Tandem mass spectrometric data analysis program used for protein identification

STAMP:

Statistical analysis of taxonomic and functional profiles

T-test:

Testing hypotheses on the mean of the normal distribution

V1–V9:

Nine hypervariable regions

TRANSTAN:

Transplant authority government of Tamil Nadu, India

HCL:

Hydrochloric acid

NaCl:

Sodium chloride

rpm:

Revolutions per minute

SDS-PAGE:

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

fm:

Femtomole

SPSS:

Statistical package for the social sciences

PPM:

Parts per million

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Acknowledgements

The corresponding author has full access to the data analysed in this study and holds all responsibilities towards the submission of this article. We acknowledge the receipt of grants from the Ganga Orthopaedic Research & Education Foundation (GOREF 2018-08) and AO Spine (AOSIN(R) 2017-04). All authors contributed equally to the preparation of this manuscript. We acknowledge the efforts taken by all the authors equally in preparing this manuscript and also thank GOREF for funding the project.

Funding

The project was mainly funded by Ganga Orthopaedic Research & Education Foundation (GOREF 2018-08) and partially by the AO Spine (AOSIN(R) 2017-04).

Author information

Authors and Affiliations

  1. Department of Spine Surgery, Ganga Hospital, 313, Mettupalayam Road, Coimbatore, India

    Shanmuganathan Rajasekaran, Dilip Chand Raja Soundararajan, K. S. Sri Vijay Anand, Ajoy Prasad Shetty & Rishi Mugesh Kanna

  2. Ganga Research Centre, No 91, Mettupalayam Road, Coimbatore, 641030, India

    Chitraa Tangavel, Monica Steffi Matchado & Sharon Miracle Nayagam

  3. Department of Plant Biotechnology, Tamil Nadu Agricultural University, Coimbatore, 641003, India

    Raveendran Muthurajan

  4. Aravind Medical Research Foundation, Madurai, 625020, India

    K. Dharmalingam

Authors
  1. Shanmuganathan Rajasekaran
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  2. Dilip Chand Raja Soundararajan
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  3. Chitraa Tangavel
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  4. Raveendran Muthurajan
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  5. K. S. Sri Vijay Anand
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  6. Monica Steffi Matchado
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  7. Sharon Miracle Nayagam
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  8. Ajoy Prasad Shetty
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  9. Rishi Mugesh Kanna
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  10. K. Dharmalingam
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Corresponding author

Correspondence to Shanmuganathan Rajasekaran.

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The study was performed only after approval of the IRB committee.

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Rajasekaran, S., Soundararajan, D.C.R., Tangavel, C. et al. Human intervertebral discs harbour a unique microbiome and dysbiosis determines health and disease. Eur Spine J 29, 1621–1640 (2020). https://doi.org/10.1007/s00586-020-06446-z

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  • DOI: https://doi.org/10.1007/s00586-020-06446-z

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