Abstract
Introduction
Low back pain (LBP) is a leading cause of disability resulting from musculoskeletal disorders among general population. The life time prevalence of single episode of low back pain in general population is reported to be around 80%. It is a leading cause of loss of productivity and the resulting economic burden due to loss of productivity is enormous. Degenerative disc disease (DDD) is a leading cause for LBP however, molecular mechanism underlying DDD are largely unknown. To date, there are few reports available on Intervertebral Disc (IVD) proteomics. Hence, weundertook the analysis of the intervertebral disc using a proteomic based approach to develop a database which may be helpful to isolate probable biomarkers and better understand the molecular basis of DDD.
Methods
Sample for proteomic analysis was obtained during lumbar discectomy in a case of intervertebral disc prolapse from a 50 year old female. IVD tissue sample was pulverizedusing liquid nitrogen and proteins were extracted. Extracted proteins were subjected to Two-Dimensional Polyacrylamide Gel Electrophoresis (2D-PAGE) analysis. Reproducible and abundantly expressed proteins were analyzed using Matrix Assisted Laser Desorption/Ionisation- Time of Flight (MALDI-TOF)Mass Spectrometry. The peptide mass fingerprinting data from the MALDI-TOF were analyzed by MASCOT online tool for the identification of proteins. Database was developed based on the results obtained from the MASCOT analysis.
Results
Thirty protein spots were identified from the degenerated IVD tissue by proteomic profiling techniques. Out of these 30 proteins, 25 proteins showed significant matches against human proteins in the NCBI database; remaining 5 spots did not generate any significant matches. The proteins which showed a significant match provide authentic evidence of the presence of these proteins in the IVD. The database was developed based on a three layer architecture model which could help investigator identify target proteins consisting of presentation layer, application layer and the data storage layer. This database would prove useful in future proteomic research into DDD.
Conclusion
Following the genomic era, molecular analysis at the proteomic level is a developing technology. This technology has the scope to identify the functions of the differently expressed proteins, from control and degenerated discs and may provide an opportunity to design novel strategies to treat degenerative disc disease in the future. Our study provides early data in the field of proteomic profiling in the clinical disease scenario of DDD.