Case Report Inhibitions of IL-1β and TNF-Α Through Scaav6-Shrnas Elevate Matrix Proteins Expressions in Degenerative Disc Cells of APOE-Knockout Rabbits
by Anja Beierfuß1, Andreas Ritsch2, Christian Kremser3, Claudius Thomé4, Demissew Shenegelegn Mern4*
1Laboratory Animal Facility, Medical University of Innsbruck, Schöpfstraße 41, A-6020 Innsbruck, Austria
2Department of Internal Medicine I, Medical University of Innsbruck, Anichstrasse 35, A-6020, Innsbruck, Austria
3Department of Radiology, Medical University of Innsbruck, Anichstrasse 35, A-6020 Innsbruck, Austria
4Department of Neurosurgery, Medical University of Innsbruck, Anichstrasse 35, A-6020 Innsbruck, Austria
*Corresponding author: Demissew Shenegelegn Mern, Department of Neurosurgery, Medical University of Innsbruck, Anichstrasse 35, A-6020 Innsbruck, Austria
Received Date: 22 July 2025
Accepted Date: 26 July 2025
Published Date: 29 July 2025
Citation: Beierfuß A, Ritsch A, Kremser C, Thome C, Mern DM. (2025). Inhibitions of IL-1β and TNF-Α Through Scaav6-Shrnas Elevate Matrix Proteins Expressions in Degenerative Disc Cells of APOE-Knockout Rabbits. Ann Case Report. 10: 2354. https://doi.org/10.29011/2574-7754.102354
Abstract
Purpose: Intervertebral disc (IVD) degeneration causing degenerative disc disease (DDD) is associated with chronic back pain. Inflammatory processes within IVDs can exacerbate progressive ECM breakdown impairing spine biomechanics. IL-1β and TNF-α are up regulated in degenerative IVDs and involved in pathological processes by promoting the expressions of inflammatory mediators. Therefore, we investigated the impacts of IL-1β and TNF-α knockdown in premature degenerative IVD cells of APOE knockout rabbits.
Methods: Knockdown was implemented using scAAV6 encoding shRNAs targeting IL-1β or TNF-α. IVD degeneration-grades were determined using MRI. Nucleus-pulposus (NP) cells were isolated from wild-type and homozygous APOE-knockout rabbits. After monolayer culture with low-glucose, cells were 3D-cultured in alginate hydrogel. MTT cell-viability assays, fluorescence microscopy, FACS, qRT-PCR, ELISA and western blotting were performed through 48 days. The viral titres, transduction efficacies (TE), cell viabilities (CV) and levels of the inflammatory, catabolic and ECM expressions were determined.
Results: The parallel knockdown of IL-1β/TNF-α showed the most effective phenotypic outcome with high TE, without any impact on CV. It additively enhanced the levels of ECM proteins by repression of matrix-metalloproteinases and aggrecanases (p < 0.001).
Conclusion: These findings could be the basis for sustainable biological treatment approaches in DDD with lower risk of immunogenicity.
Keywords: Inflammatory cytokines; Catabolic enzymes; Extracellular matrix breakdown; ScAAV6-shRNA mediated knockdown of IL-1β and TNF-α; Biological treatment of degenerative disc disease
© by the Authors & Gavin Publishers. This is an Open Access Journal Article Published Under Attribution-Share Alike CC BY-SA: Creative Commons Attribution-Share Alike 4.0 International License. Read More About Open Access Policy.