Lentivirus Is an Efficient and Stable Transduction Vector for Intervertebral Disc Cells

Area of Science:

• Biotechnology
• Gene Therapy
• Cell Biology

Background:

• Degenerative disc disease (DDD) is a significant cause of low back pain.
• Current treatments for DDD are limited, highlighting the need for novel therapeutic approaches.
• Gene therapy offers a potential strategy for regenerating intervertebral disc (IVD) cells.

Purpose of the Study:

• To assess the efficacy and durability of lentiviral vector transduction in both in vitro and in vivo models of intervertebral disc cells.
• To determine the optimal multiplicity of infection (MOI) for lentiviral transduction of disc cells.
• To evaluate the safety and gene expression persistence following in vivo lentiviral delivery to rat IVDs.

Main Methods:

• Human nucleus pulposus and anulus fibrosus cells were cultured and transduced with lentivirus carrying enhanced green fluorescent protein (eGFP) at various MOIs.
• Cell viability was assessed using the trypan blue exclusion test, and transduction efficiency was quantified by fluorescence-activated cell sorting (FACS).
• In vivo studies involved injecting lentivirus into rat IVDs, with eGFP expression analyzed at multiple time points post-transduction.

Main Results:

• Lentiviral transduction did not compromise intervertebral disc cell viability or growth.
• An optimal transduction efficiency of 97% was achieved at an MOI of 60, irrespective of patient or disease characteristics.
• In vivo, eGFP expression peaked at 7 days and was detectable up to 28 days post-transduction, with no observed adverse systemic effects.

Conclusions:

• Lentiviral vectors demonstrate high efficiency and stability for transducing intervertebral disc cells.
• This vector system holds significant potential as a gene therapy tool for managing human degenerative disc disease.
• Further research into lentiviral vector applications could lead to advanced treatments for spinal degeneration.