Conditioned medium from intervertebral disc inhibits autologous mesenchymal stromal cells and osteoblasts

Chen, Shuimu; Croft, Andreas S.; Bigdon, Sebastian; Li, Zhen; Oberli, Andrea E.; Crump, Katherine B.; Gantenbein, Benjamin (2023). Conditioned medium from intervertebral disc inhibits autologous mesenchymal stromal cells and osteoblasts. In: Annual Meeting of the Swiss Society of Biomedical Engineerin. 13 September.

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Introduction

Low back pain (LBP) is a significant global burden and is associated with the degeneration of the spine and human intervertebral discs (hIVD) [1]. Current “gold standard” treatment for hIVD degeneration is the removal of the affected hIVD tissue and to promote spinal fusion. However, in some cases this process fail because of non-fusions after operation [2,3]. One possible reason is that bone morphogenetic protein (BMP) antagonists i.e., are secreted by the IVD at higher doses, which has a inhibitory effect of osteogenesis in the presence of IVD cells [4].

The purpose of this study is to explore the effect of conditioned medium (CM) from hIVD on autologous osteoblasts (hOBs) and bone marrow mesenchymal stem cells (hMSCs).

Material and Methods

Collect the bone fragments, bone marrow and intervertebral disc from the same patients after spinal operation. Isolation of hOB, hMSC from bone and bone marrow, as well as three main cell types from hIVD, nucleus pulposus (hNP), annulus fibrosus (hAF) and cartilage endplate (hCEP). Make alginate beads with hIVDs and collect the conditioned medium. Conditioned medium is used to culture hOBs and hMSCs. Evaluate the osteogenesis of hOBs and hMSC, as well as potential cause.

Results

Figure 1. ALP expression significantly decreases on Day 10 when adding CM into hMSC and hOB. (pvalue, * < 0.05; * * < 0.01; * * * < 0.001; * * * *

Figure 2. A trend of lower calcium deposition could be observed on Day 21 using alizarin red staining after hOB and hMSC cultured with CM from different hIVD cells. (Calcium deposits: Bright orange red; hMSC&

hOB, N=3-5)

Figure 3. The expression of Noggin is mainly higher in all cell types of hIVD compared with hOB, as well as hMSC. (hMSC& hOB, shown are the means, N=3-5)

Conclusions and Discussion

➢ ➢ ➢

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Noggin is higher expressed in hIVD cells compared to hOB and hMSC (Fig 3).

The osteogenesis of hOBs and hMSCs is generally inhibited in the presence of CM from hIVD cells. The high expression level of Noggin in hIVD cells may be a potential inhibitor of autologous OBs and MSCs.

Knockdown of Noggin in hIVD cells could improve the osteogenesis of hOBs.

References

1. James et al, The Lancet, 392(10159): 1789-1858, 2018.

2. Serra et al, Biofabrication, 8(3): 035001, 2016.

3. Watkins et al, Medicine, 93(29): e275, 2014.

4. Chan et al, Arthritis Research & Therapy, 18:29, 2016.

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