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Degenerate human nucleus pulposus cells promote neurite outgrowth in neural cells.

Richardson SM, Purmessur D, Baird P, Probyn B, Freemont AJ, Hoyland JA - PLoS ONE (2012)

Bottom Line: Co-culture of non-degenerate NP cells with neural cells resulted in both an inhibition of neurite outgrowth and reduction in percentage of neurite expressing cells.Conversely co-culture with degenerate NP cells resulted in an increase in both neurite length and percentage of neurite expressing cells.Addition of anti-NGF to the co-culture with degenerate cells resulted in a decrease in percentage of neurite expressing cells, while addition of anti-BDNF resulted in a decrease in both neurite length and percentage of neurite expressing cells.

View Article: PubMed Central - PubMed

Affiliation: Centre for Regenerative Medicine, Institute of Inflammation and Repair, Faculty of Medical and Human Sciences, The University of Manchester, Manchester, United Kingdom.

ABSTRACT
Innervation of nociceptive nerve fibres into the normally aneural nucleus pulposus (NP) of the intervertebral disc (IVD) occurs during degeneration resulting in discogenic back pain. The neurotrophins nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), which are associated with stimulation of axonal outgrowth and nociception by neuronal cells, are both expressed by NP cells, with BDNF levels increasing with disease severity. However the mechanism of interaction between human NP cells and neural cells has yet to be fully elucidated. Therefore the aim of this study was to determine whether non-degenerate or degenerate human NP cells inhibit or stimulate neural outgrowth and whether any outgrowth is mediated by NGF or BDNF. Human NP cells from non-degenerate and degenerate IVD were cultured in alginate beads then co-cultured for 48 hours with human SH-SY5Y neuroblastoma cells. Co-culture of non-degenerate NP cells with neural cells resulted in both an inhibition of neurite outgrowth and reduction in percentage of neurite expressing cells. Conversely co-culture with degenerate NP cells resulted in an increase in both neurite length and percentage of neurite expressing cells. Addition of anti-NGF to the co-culture with degenerate cells resulted in a decrease in percentage of neurite expressing cells, while addition of anti-BDNF resulted in a decrease in both neurite length and percentage of neurite expressing cells. Our findings show that while non-degenerate NP cells are capable of inhibiting neurite outgrowth from human neural cells, degenerate NP cells stimulate outgrowth. Neurotrophin blocking studies demonstrated that both NGF and BDNF, secreted by degenerate NP cells, may play a role in this stimulation with BDNF potentially playing the predominant role. These findings suggest that NP cells are capable of regulating nerve ingrowth and that neoinnervation occurring during IVD degeneration may be stimulated by the NP cells themselves.

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Analysis of neural cell behaviour following co-culture with normal and degenerate human NP cells.Histograms illustrating the percentage number of neurite expressing cells (A and C) and mean neurite length (B and D) from differentiated SH-SY5Y cells co-cultured with non-degenerate (A and B) and degenerate (C and D) NP cells without contact for 48 hours (n = 3 non-degenerate, n = 3 degenerate; *  = P<0.05).
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pone-0047735-g003: Analysis of neural cell behaviour following co-culture with normal and degenerate human NP cells.Histograms illustrating the percentage number of neurite expressing cells (A and C) and mean neurite length (B and D) from differentiated SH-SY5Y cells co-cultured with non-degenerate (A and B) and degenerate (C and D) NP cells without contact for 48 hours (n = 3 non-degenerate, n = 3 degenerate; *  = P<0.05).

Mentions: After 48 hours co-culture of SH-SY5Y cells with cells derived from non-degenerate discs there was a significant (P = 0.0002) decrease in the percentage number of neurite expressing SH-SY5Y cells in the co-cultured population (31.3%) compared to SH-SY5Y cells cultured alone (45.2%)(figure 3A). When examining mean neurite length a decrease was also observed with SH-SY5Y cells co-cultured with non-degenerate NP cells (29.9 µm) when compared to its corresponding control (31.7 µm), although this decrease was not significant (P = 0.25)(figure 3B).


Degenerate human nucleus pulposus cells promote neurite outgrowth in neural cells.

Richardson SM, Purmessur D, Baird P, Probyn B, Freemont AJ, Hoyland JA - PLoS ONE (2012)

Analysis of neural cell behaviour following co-culture with normal and degenerate human NP cells.Histograms illustrating the percentage number of neurite expressing cells (A and C) and mean neurite length (B and D) from differentiated SH-SY5Y cells co-cultured with non-degenerate (A and B) and degenerate (C and D) NP cells without contact for 48 hours (n = 3 non-degenerate, n = 3 degenerate; *  = P<0.05).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3472988&req=5

pone-0047735-g003: Analysis of neural cell behaviour following co-culture with normal and degenerate human NP cells.Histograms illustrating the percentage number of neurite expressing cells (A and C) and mean neurite length (B and D) from differentiated SH-SY5Y cells co-cultured with non-degenerate (A and B) and degenerate (C and D) NP cells without contact for 48 hours (n = 3 non-degenerate, n = 3 degenerate; *  = P<0.05).
Mentions: After 48 hours co-culture of SH-SY5Y cells with cells derived from non-degenerate discs there was a significant (P = 0.0002) decrease in the percentage number of neurite expressing SH-SY5Y cells in the co-cultured population (31.3%) compared to SH-SY5Y cells cultured alone (45.2%)(figure 3A). When examining mean neurite length a decrease was also observed with SH-SY5Y cells co-cultured with non-degenerate NP cells (29.9 µm) when compared to its corresponding control (31.7 µm), although this decrease was not significant (P = 0.25)(figure 3B).

Bottom Line: Co-culture of non-degenerate NP cells with neural cells resulted in both an inhibition of neurite outgrowth and reduction in percentage of neurite expressing cells.Conversely co-culture with degenerate NP cells resulted in an increase in both neurite length and percentage of neurite expressing cells.Addition of anti-NGF to the co-culture with degenerate cells resulted in a decrease in percentage of neurite expressing cells, while addition of anti-BDNF resulted in a decrease in both neurite length and percentage of neurite expressing cells.

View Article: PubMed Central - PubMed

Affiliation: Centre for Regenerative Medicine, Institute of Inflammation and Repair, Faculty of Medical and Human Sciences, The University of Manchester, Manchester, United Kingdom.

ABSTRACT
Innervation of nociceptive nerve fibres into the normally aneural nucleus pulposus (NP) of the intervertebral disc (IVD) occurs during degeneration resulting in discogenic back pain. The neurotrophins nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), which are associated with stimulation of axonal outgrowth and nociception by neuronal cells, are both expressed by NP cells, with BDNF levels increasing with disease severity. However the mechanism of interaction between human NP cells and neural cells has yet to be fully elucidated. Therefore the aim of this study was to determine whether non-degenerate or degenerate human NP cells inhibit or stimulate neural outgrowth and whether any outgrowth is mediated by NGF or BDNF. Human NP cells from non-degenerate and degenerate IVD were cultured in alginate beads then co-cultured for 48 hours with human SH-SY5Y neuroblastoma cells. Co-culture of non-degenerate NP cells with neural cells resulted in both an inhibition of neurite outgrowth and reduction in percentage of neurite expressing cells. Conversely co-culture with degenerate NP cells resulted in an increase in both neurite length and percentage of neurite expressing cells. Addition of anti-NGF to the co-culture with degenerate cells resulted in a decrease in percentage of neurite expressing cells, while addition of anti-BDNF resulted in a decrease in both neurite length and percentage of neurite expressing cells. Our findings show that while non-degenerate NP cells are capable of inhibiting neurite outgrowth from human neural cells, degenerate NP cells stimulate outgrowth. Neurotrophin blocking studies demonstrated that both NGF and BDNF, secreted by degenerate NP cells, may play a role in this stimulation with BDNF potentially playing the predominant role. These findings suggest that NP cells are capable of regulating nerve ingrowth and that neoinnervation occurring during IVD degeneration may be stimulated by the NP cells themselves.

Show MeSH
Related in: MedlinePlus