A Well-Controlled Nucleus Pulposus Tissue Culture System with Injection Port for Evaluating Regenerative Therapies

Arkesteijn, I. T. M.; Mouser, Vivian H. M.; Mwale, Fackson; Dijk, Bart G. M. van; Ito, Keita

doi:10.1007/s10439-015-1428-y

Cited by 7 publications

(9 citation statements)

References 36 publications

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“…15 Improved NP culture systems are currently available for future experiments. 29,31 Although the results from this study show that direct injections of NCs are not effective for NP regeneration, this finding should be interpreted with care for translation to the human situation. First of all, the NP tissue used in this study corresponded to human grade II on the Thompson grading scale.…”

Section: Discussionmentioning

confidence: 73%

The Regenerative Potential of Notochordal Cells in a Nucleus Pulposus Explant

Arkesteijn

Potier

Ito

2017

Global Spine Journal

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Study DesignIn vitro disk explant culture.ObjectiveNotochordal cells (NCs) have been shown to upregulate matrix production by nucleus pulposus (NP) cells in coculture. To examine the translation of these in vitro results to a nativelike setting, the regenerative potential of NCs injected into NP tissue was assessed in this study.MethodsNP explants were cultured after injection with NCs in phosphate-buffered saline (PBS) or with PBS alone (sham). At days 0 and 42, cell viability and morphology, water, DNA, sulfated glycosaminoglycan and hydroxyproline content, and gene expression of anabolic markers were analyzed.ResultsNCs remained viable during culture, but their morphology changed. The biochemical content remained unchanged, except for the DNA content in the NC group. Overall ACAN expression remained unchanged, whereas COL2A1 decreased during culture.ConclusionsNo overall anabolic response was observed when NCs were injected into NP explants. NCs were found to survive but did not display the typical NC morphology by the end of the culture period.

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Section: Discussionmentioning

confidence: 73%

The Regenerative Potential of Notochordal Cells in a Nucleus Pulposus Explant

Arkesteijn

Potier

Ito

2017

Global Spine Journal

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“…Our study, however, suggests that continuous HP loading (repeating cyclic followed by constant HP) is required. This contradictory suggestion may be related to another important mechanical factor, confinement 23,31 . For example, during off‐loading, the NP absorbs fluid, generating swelling pressure.…”

Section: Discussionmentioning

confidence: 99%

In vitro nucleus pulposus tissue model with physicochemical stresses

2020

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Intervertebral discs (IVDs) are exposed to changes in physicochemical stresses including hydrostatic and osmotic pressure via diurnal spinal motion. Homeostasis, degeneration, and regeneration in IVDs have been studied using in vitro, ex vivo, and animal models. However, incubation of nucleus pulposus (NP) cells in medium has limited capability to reproduce anabolic turnover and regeneration under physicochemical stresses. We developed a novel pressure/perfusion cell culture system and a semipermeable membrane pouch device for enclosing isolated NP cells for in vitro incubation under physicochemical stresses. We assessed the performance of this system to identify an appropriate stress loading regimen to promote gene expression and consistent accumulation of extracellular matrices by bovine caudal NP cells. Cyclic hydrostatic pressure (HP) for 4 days followed by constant HP for 3 days in high osmolality (HO; 450 mOsm/kg H 2 O) showed a trend towards upregulated aggrecan expression and dense accumulation of keratan sulfate without gaps by the NP cells. Furthermore, a repetitive regimen of cyclic HP for 2 days followed by constant HP for 1 day in HO (repeated twice) significantly upregulated gene expression of aggrecan (P < .05) compared to no pressure and suppressed matrix metalloproteinase-13 expression (P < .05) at 6 days. Our culture system and pouches will be useful to reproduce physicochemical stresses in NP cells for simulating anabolic, catabolic, and homeostatic turnover under diurnal spinal motion.

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“…Medium osmolarity (346 mOsmol/L) was not artificially increased as NPs were cultured in confinement. To prevent swelling after the NP is released from its confined space between the vertebrae and encircled by the AF, we developed an NP bioreactor chamber based on a previous study 19 . This bioreactor chamber allows axial loading, mechanical testing, and contains a port for central injection (Figure 2).…”

Section: Methodsmentioning

confidence: 99%

A bovine nucleus pulposus explant culture model

Salzer

Mouser

Tryfonidou

et al. 2021

Journal Orthopaedic Research

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Low back pain is a global health problem that is frequently caused by intervertebral disc degeneration (IVDD). Sulfated glycosaminoglycans (sGAGs) give the healthy nucleus pulposus (NP) a high fixed charge density (FCD), which creates an osmotic pressure that enables the disc to withstand high compressive forces. However, during IVDD sGAG reduction in the NP compromises biomechanical function. The aim of this study was to develop an ex vivo NP explant model with reduced sGAG content and subsequently investigate biomechanical restoration via injection of proteoglycan‐containing notochordal cell‐derived matrix (NCM). Bovine coccygeal NP explants were cultured in a bioreactor chamber and sGAG loss was induced by chondroitinase ABC (chABC) and cultured for up to 14 days. Afterwards, diurnal loading was studied, and explant restoration was investigated via injection of NCM. Explants were analyzed via histology, biochemistry, and biomechanical testing via stress relaxation tests and height measurements. ChABC injection induced dose‐dependent sGAG reduction on Day 3, however, no dosing effects were detected after 7 and 14 days. Diurnal loading reduced sGAG loss after injection of chABC. NCM did not show an instant biomechanical (equilibrium pressure) or biochemical (FCD) restoration, as the injected fixed charges leached into the medium, however, NCM stimulated proliferation and increased Alcian blue staining intensity and matrix organization. NCM has biological repair potential and biomaterial/NCM combinations, which could better entrap NCM within the NP tissue, should be investigated in future studies. Concluding, chABC induced progressive, time‐, dose‐ and loading‐dependent sGAG reduction that led to a loss of biomechanical function. Keywords biomechanics | intervertebral disc | matrix degradation | low back pain | proteoglycans

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A Well-Controlled Nucleus Pulposus Tissue Culture System with Injection Port for Evaluating Regenerative Therapies

Cited by 7 publications

References 36 publications

The Regenerative Potential of Notochordal Cells in a Nucleus Pulposus Explant

The Regenerative Potential of Notochordal Cells in a Nucleus Pulposus Explant

In vitro nucleus pulposus tissue model with physicochemical stresses

A bovine nucleus pulposus explant culture model

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