Inhibition of proteoglycan and type II collagen synthesis of disc nucleus cells by nicotine

Abstract

<jats:p content-type="fine-print"><jats:italic>Object.</jats:italic> Systemic nicotine has been hypothesized to cause degeneration of the intervertebral disc which in turn decreases vascular supply to the disc through a cholinergic receptor—mediated process. Another possible mechanism may be through direct regulatory effects on disc cells. In this study, the authors tested the hypothesis that nicotine adversely affects nucleus pulposus cells by directly inhibiting proteoglycan synthesis and gene expression of type II collagen (Phase I study). They also assessed the hypothesis that nicotine inhibits the bone morphogenetic protein (BMP)—2-induced upregulation of extracellular matrix (Phase II study).</jats:p> <jats:p content-type="fine-print"><jats:italic>Methods.</jats:italic> Cells were isolated from nucleus pulposus obtained in rat lumbar discs and cultured on a monolayer. Media were treated with nicotine and/or recombinant human (rh)BMP-2 for 7 days. Sulfated glycosaminoglycan (SO<jats:sub>4</jats:sub>-GAG) in media was quantified using 1,9-dimethylmethylene blue (DMMB) assay. Gene assay of types I and II collagen, Sox9, and glyceraldehyde-3-phosphate dehydrogenase were quantified using reverse transcriptase—polymerase chain reaction (RT-PCR) and real time PCR. In the Phase I study, nicotine-treated (100 µg/ml) and nontreated cells were compared. The s-GAG production and messenger RNA (mRNA) of type II collagen and Sox9 decreased significantly in the nicotine-treated group. In the Phase II study, five groups were compared: 1) nontreatment; 2) rhBMP-2 only (100 ng/ml); and 3–5) with rhBMP-2 (100 ng/ml) and increasing doses of nicotine (1 [third group], 10, [fourth group], 100 [fifth group] µg/ml). The SO<jats:sub>4</jats:sub>-GAG production and mRNA of type II collagen and Sox9 decreased significantly in the groups treated with rhBMP-2 combined with 10 and 100 µg/ml of nicotine compared with the group treated with rhBMP-2.</jats:p> <jats:p content-type="fine-print"><jats:italic>Conclusions.</jats:italic> The results of this study raise the possibility that nicotine may contribute to the process of disc degeneration by a direct effect on the nucleus pulposus cells, possibly by antagonizing the effect of BMP-2.</jats:p>

Alteration of Gene Expression in Intervertebral Disc Degeneration of Passive Cigarette- Smoking Rats: Separate Quantitation in Separated Nucleus pulposus and Annulus fibrosus

“…We have previously shown that nicotine could cause intervertebral disc degeneration by reducing the blood fl ow to intervertebral discs [4] . On the other hand, Kim et al [3] showed that nicotine, by directly affecting nucleus pulposus (NP) cells in the intervertebral discs, is involved in intervertebral disc degeneration by inducing the synthesis of sulfated glycosaminoglycans, and downregulating type II collagen and Sox9 mRNAs via inhibition of the effect of bone morphogenic protein-2. However, cigarettes contain a variety of constituents, and at least two or more factors seem to be involved in intervertebral disc degeneration.…”

“…Recent in vivo experiments have demonstrated that exposure to nicotine, a major chemical constituent of cigarettes, induces intervertebral disc degeneration [2][3][4][5] . Exposure to nicotine, through systemic effects related to cholinergic nicotinic receptors, leads to increased blood pressure, vasoconstriction and vascular resistance.…”

MicroRNA-125b-1-3p mediates intervertebral disc degeneration in rats by targeting teashirt zinc finger homeobox 3

“…It had been demonstrated that H 2 O 2 may stimulate DNA oxidative damage in the NP cells of rats (24) and in the present research a DNA oxidative damage model was established by applying different concentrations of H 2 O 2 (0.05, 0.1 and 0.2 mM/l). COL2 expression was used to monitor the degeneration of NP cells, following a previously described protocol (25). The expression of COL2 decreased as increasing concentrations of H 2 O 2 were used, whereas the expression of TSHZ3 increased in an H 2 O 2 concentration-dependent manner (Fig.…”

Beeinflusst Rauchen Rückenschmerzen und spinale Operationsergebnisse?

“…Durch Zigarettenrauch verursachte Auswirkungen auf die Bandscheiben wurden von Holm und Nachemson erforscht, welche insbesondere vaskuläre Effekte und erhöhte Kohlenstoffmonoxidkonzentrationen für die Bandscheibendegeneration verantwortlich machen [8]. Während niedrige (100 µg/mL [27]) und sehr hohe Nikotindosen von etwa 8000-50 000 µg/mL [17] den Metabolismus von Zellen des Nucleus pulposus sowie die Produktion von Glycosaminoglykanen und Kollagen in vitro hemmen, werden diese durch mittelhohe Dosen (etwa 4000 µg/mL) vorübergehend stimuliert, bis sie nach 7 Tagen ebenfalls abnehmen [17]. Die Serumkonzentration von Nikotin bei starken Rauchern (20-30 Zigaretten pro Tag) beträgt etwa 0,01-0,07 µg/ml [28] und liegt somit z. T. deutlich unter den applizierten Konzentrationen der obigen In-vitro-Versuche.…”