Abstract
In vitro evaluation of nucleus pulposus (NP) tissue regeneration would be useful, but current systems for NP culture are not ideal for injections. The aim of this study was to develop a long-term culture system for NP tissue that allows injections of regenerative agents. Bovine caudal NPs were harvested and placed in the newly designed culture system. After equilibration of the tissue to 0.3 MPa the volume was fixed and the tissue was cultured for 28 days. The cell viability and extracellular matrix composition remained unchanged during the culture period and gene expression profiles were similar to those obtained in earlier studies. Furthermore, to test the responsiveness of bovine caudal NPs in the system, samples were cultured for 4 days and injected twice (day 1 and 3) with (1) PBS, (2) Link-N, for regeneration, and (3) TNF-α, for degeneration. It was shown that TNF-α increased COX2 gene expression, whereas no effect of Link-N was detected. In conclusion, the newly designed system allows long-term culture of NP tissue, wherein tissue reactions to injected stimulants can be observed.
Highlights
Intervertebral discs support high magnitude loads and allow multi-directional flexibility in the spine, due to an ingenious interplay of the tissues within the disc
This article is published with open access at Springerlink.com often requires long-term studies, we focused on optimizing currently available nucleus pulposus (NP) culture systems
NP tissue biopsies were placed in the culture system and equilibrated
Summary
Intervertebral discs support high magnitude loads and allow multi-directional flexibility in the spine, due to an ingenious interplay of the tissues within the disc. The most widely used model to test regenerative therapies is the in vivo animal model. In most test cases, rodents or other small animals are used in which degeneration is artificially induced. First of all, such small animals have a mature nuclear cell type, which is different than that of humans.[1] Second, the artificially induced degeneration is different from the human pathophysiological degenerative cascade. Translation of results obtained in studies with artificial degeneration to naturally degenerated human NPs is difficult.[2] Testing regenerative therapies on in vitro cultured cells is another standard method. Cell metabolism and phenotype outside of the native matrix and environment change.[9] tissue culture models are of great interest
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