Limits...
Characterization of metabolites determined by means of 1H HR MAS NMR in intervertebral disc degeneration.

Pacholczyk-Sienicka B, Radek M, Radek A, Jankowski S - MAGMA (2014)

Bottom Line: The concentrations of 2-propanol and lactate increase with age.PCA analysis of ex vivo (1)H HR MAS NMR data revealed the occurrence of two groups: healthy and degenerative disc tissues.The effects of insufficient nutrient supply of discs, leading to their degeneration and back pain, are discussed.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Chemistry, Institute of Organic Chemistry, Łódź University of Technology, Lodz, Poland.

ABSTRACT

Object: The objective of this study is the identification of metabolites by means of (1)H high resolution magic angle spinning nuclear magnetic resonance ((1)H HR MAS NMR) spectroscopy and the evaluation of their applicability in distinguishing between healthy and degenerated disc tissues.

Materials and methods: Differences between the metabolic profiles of healthy and degenerated disc tissues were studied by means of (1)H HR MAS NMR. Analysis was performed for 81 disc tissue samples (control samples n = 21, degenerated disc tissue samples n = 60). Twenty six metabolites (amino acids, carbohydrates, and alcohols) were identified and quantified.

Results: The results indicate that the metabolic profile of degenerated discs is characterized by the presence of 2-propanol and the absence of scyllo-inositol and taurine. The concentrations of 2-propanol and lactate increase with age.

Conclusion: PCA analysis of ex vivo (1)H HR MAS NMR data revealed the occurrence of two groups: healthy and degenerative disc tissues. The effects of insufficient nutrient supply of discs, leading to their degeneration and back pain, are discussed.

Show MeSH

Related in: MedlinePlus

Representative ex vivo 1H HR MAS NMR spectra of a healthy disc (a), and of a degenerated disc for the aliphatic (b) and aromatic (c) regions of the spectrum
© Copyright Policy - OpenAccess
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4385564&req=5

Fig1: Representative ex vivo 1H HR MAS NMR spectra of a healthy disc (a), and of a degenerated disc for the aliphatic (b) and aromatic (c) regions of the spectrum

Mentions: Representative ex vivo 1H HR MASNMR spectra of healthy and degenerated disc tissues are shown in Fig. 1. Assignment of metabolite resonances was based on analysis of one dimensional 1H, two dimensional correlation spectroscopy (COSY), and total correlation spectroscopy (TOCSY) NMR spectra. The collected data were compared with those from the literature [25] and with the spectra of metabolites recorded on a Bruker Avance II Plus700 MHz spectrometer. The1H HR MAS NMR spectra for nucleus pulposus and annulus fibrosus specimens were very similar (see Figure SM1 in supplementary material). In the HR MAS spectra of disc tissues, signals of five groups of compounds were observed. The main metabolites characterizing disc tissues are labeled in Fig. 1 and the pool of metabolites is reported in Table 1. The largest group consisted of 12 amino acids, which were derived from protein cores and were also formed by collagen breakdown. The second group of compounds was comprised of carboxylic acids and their derivatives, mainly lactic acid, citric acid, formate, acetate, and acetone. Moreover, some carbohydrates (mainly α- and β-glucose and chondroitin sulfate) and pyrimidine derivatives (e.g. uracil) were found in the spectra of disc tissues. Alcohols, such as myo-inositol, scyllo-inositol, and 2-propanol, were also identified. In the case of 2-propanol, the possibility of its exogenous origin was checked at every step from surgery to HR MAS measurements, but it was deemed possible for 2-propanol to have been introduced into disc tissue samples as a result of its antiseptic use. The patients were being administered pain relief and non-steroidal anti-inflammatory drugs, but 2-propanol was not found to be a component or metabolite of those drugs. Resonances of 2-propanol was observed due to good resolution of 1H NMR spectra and confirmed by COSY/TOCSY experiments (see Figure SM3–SM4 in supplementary materials). The unassigned signal at 1.2 ppm reported earlier [2] probably corresponds to 2-propanol, but was not recognized in poorly resolved spectra.Fig. 1


Characterization of metabolites determined by means of 1H HR MAS NMR in intervertebral disc degeneration.

Pacholczyk-Sienicka B, Radek M, Radek A, Jankowski S - MAGMA (2014)

Representative ex vivo 1H HR MAS NMR spectra of a healthy disc (a), and of a degenerated disc for the aliphatic (b) and aromatic (c) regions of the spectrum
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4385564&req=5

Fig1: Representative ex vivo 1H HR MAS NMR spectra of a healthy disc (a), and of a degenerated disc for the aliphatic (b) and aromatic (c) regions of the spectrum
Mentions: Representative ex vivo 1H HR MASNMR spectra of healthy and degenerated disc tissues are shown in Fig. 1. Assignment of metabolite resonances was based on analysis of one dimensional 1H, two dimensional correlation spectroscopy (COSY), and total correlation spectroscopy (TOCSY) NMR spectra. The collected data were compared with those from the literature [25] and with the spectra of metabolites recorded on a Bruker Avance II Plus700 MHz spectrometer. The1H HR MAS NMR spectra for nucleus pulposus and annulus fibrosus specimens were very similar (see Figure SM1 in supplementary material). In the HR MAS spectra of disc tissues, signals of five groups of compounds were observed. The main metabolites characterizing disc tissues are labeled in Fig. 1 and the pool of metabolites is reported in Table 1. The largest group consisted of 12 amino acids, which were derived from protein cores and were also formed by collagen breakdown. The second group of compounds was comprised of carboxylic acids and their derivatives, mainly lactic acid, citric acid, formate, acetate, and acetone. Moreover, some carbohydrates (mainly α- and β-glucose and chondroitin sulfate) and pyrimidine derivatives (e.g. uracil) were found in the spectra of disc tissues. Alcohols, such as myo-inositol, scyllo-inositol, and 2-propanol, were also identified. In the case of 2-propanol, the possibility of its exogenous origin was checked at every step from surgery to HR MAS measurements, but it was deemed possible for 2-propanol to have been introduced into disc tissue samples as a result of its antiseptic use. The patients were being administered pain relief and non-steroidal anti-inflammatory drugs, but 2-propanol was not found to be a component or metabolite of those drugs. Resonances of 2-propanol was observed due to good resolution of 1H NMR spectra and confirmed by COSY/TOCSY experiments (see Figure SM3–SM4 in supplementary materials). The unassigned signal at 1.2 ppm reported earlier [2] probably corresponds to 2-propanol, but was not recognized in poorly resolved spectra.Fig. 1

Bottom Line: The concentrations of 2-propanol and lactate increase with age.PCA analysis of ex vivo (1)H HR MAS NMR data revealed the occurrence of two groups: healthy and degenerative disc tissues.The effects of insufficient nutrient supply of discs, leading to their degeneration and back pain, are discussed.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Chemistry, Institute of Organic Chemistry, Łódź University of Technology, Lodz, Poland.

ABSTRACT

Object: The objective of this study is the identification of metabolites by means of (1)H high resolution magic angle spinning nuclear magnetic resonance ((1)H HR MAS NMR) spectroscopy and the evaluation of their applicability in distinguishing between healthy and degenerated disc tissues.

Materials and methods: Differences between the metabolic profiles of healthy and degenerated disc tissues were studied by means of (1)H HR MAS NMR. Analysis was performed for 81 disc tissue samples (control samples n = 21, degenerated disc tissue samples n = 60). Twenty six metabolites (amino acids, carbohydrates, and alcohols) were identified and quantified.

Results: The results indicate that the metabolic profile of degenerated discs is characterized by the presence of 2-propanol and the absence of scyllo-inositol and taurine. The concentrations of 2-propanol and lactate increase with age.

Conclusion: PCA analysis of ex vivo (1)H HR MAS NMR data revealed the occurrence of two groups: healthy and degenerative disc tissues. The effects of insufficient nutrient supply of discs, leading to their degeneration and back pain, are discussed.

Show MeSH
Related in: MedlinePlus