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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 degenerate human NP cells with or without addition of anti-BDNF antibody.Histograms illustrating the percentage number of neurite expressing cells (A) and mean neurite length (B) from differentiated SH-SY5Y cells co-cultured with degenerate NP cells without contact for 48 hours with or without addition of anti-BDNF antibody (n = 4; *  = P<0.05).
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pone-0047735-g005: Analysis of neural cell behaviour following co-culture with degenerate human NP cells with or without addition of anti-BDNF antibody.Histograms illustrating the percentage number of neurite expressing cells (A) and mean neurite length (B) from differentiated SH-SY5Y cells co-cultured with degenerate NP cells without contact for 48 hours with or without addition of anti-BDNF antibody (n = 4; *  = P<0.05).

Mentions: Co-cultures of SH-SY5Y cells and cells derived from degenerate discs were also established to test the effect of anti-BDNF in co-cultures on neurite outgrowth. Again this cohort showed similar trends to the original co-culture experiments, with significant increases in both percentage number of neurite expressing cells (71% in SH-SY5Y cells cultured alone, compared to 84% in co-cultured cells, P = 0.0054; figure 5A) and mean neurite length (34.2 µm in controls compared to 39.7 µm in co-cultured cells, P<0.0001; figure 5B). Following inclusion of anti-BDNF during co-culture the percentage number of neurite expressing cells decreased significantly (P<0.0001) to 70%, in line with control cells cultured alone. Furthermore, mean neurite length also decreased significantly (P<0.0001) to approximately those seen in control samples (33.5 µm) following addition of anti-BDNF antibody.


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 degenerate human NP cells with or without addition of anti-BDNF antibody.Histograms illustrating the percentage number of neurite expressing cells (A) and mean neurite length (B) from differentiated SH-SY5Y cells co-cultured with degenerate NP cells without contact for 48 hours with or without addition of anti-BDNF antibody (n = 4; *  = P<0.05).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0047735-g005: Analysis of neural cell behaviour following co-culture with degenerate human NP cells with or without addition of anti-BDNF antibody.Histograms illustrating the percentage number of neurite expressing cells (A) and mean neurite length (B) from differentiated SH-SY5Y cells co-cultured with degenerate NP cells without contact for 48 hours with or without addition of anti-BDNF antibody (n = 4; *  = P<0.05).
Mentions: Co-cultures of SH-SY5Y cells and cells derived from degenerate discs were also established to test the effect of anti-BDNF in co-cultures on neurite outgrowth. Again this cohort showed similar trends to the original co-culture experiments, with significant increases in both percentage number of neurite expressing cells (71% in SH-SY5Y cells cultured alone, compared to 84% in co-cultured cells, P = 0.0054; figure 5A) and mean neurite length (34.2 µm in controls compared to 39.7 µm in co-cultured cells, P<0.0001; figure 5B). Following inclusion of anti-BDNF during co-culture the percentage number of neurite expressing cells decreased significantly (P<0.0001) to 70%, in line with control cells cultured alone. Furthermore, mean neurite length also decreased significantly (P<0.0001) to approximately those seen in control samples (33.5 µm) following addition of anti-BDNF antibody.

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