Adipose-derived stromal cells protect intervertebral disc cells in compression: implications for stem cell regenerative disc therapy

INTRODUCTION: Abnormal biomechanics plays a role in intervertebral disc degeneration. Adipose-derived stromal cells (ADSCs) have been implicated in disc integrity; however, their role in the setting of mechanical stimuli upon the disc's nucleus pulposus (NP) remains unknown. As such, the present study aimed to evaluate the influence of ADSCs upon NP cells in compressive load culture. METHODS: Human NP cells were cultured in compressive load at 3.0MPa for 48 hours with or without ADSCs co-culture (the ratio was 50:50). We used flow cytometry, live/dead staining and scanning electron microscopy (SEM) to evaluate cell death, and determined the expression of specific apoptotic pathways by characterizing the expression of activated caspases-3, -8 and -9. We further used real-time (RT-) PCR and immunostaining to determine the expression of the extracellular matrix (ECM), mediators of matrix degradation (e.g. MMPs, TIMPs and ADAMTSs), pro-inflammatory factors and NP cell phenotype markers. RESULTS: ADSCs inhibited human NP cell apoptosis via suppression of activated caspase-9 and caspase-3. Furthermore, ADSCs protected NP cells from the degradative effects of compressive load by significantly up-regulating the expression of ECM genes (SOX9, COL2A1 and ACAN), tissue inhibitors of metalloproteinases (TIMPs) genes (TIMP-1 and TIMP-2) and cytokeratin 8 (CK8) protein expression. Alternatively, ADSCs showed protective effect by inhibiting compressive load mediated increase of matrix metalloproteinases (MMPs; MMP-3 and MMP-13), disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs; ADAMTS-1 and 5), and pro-inflammatory factors (IL-1beta, IL-6, TGF-beta1 and TNF-alpha). CONCLUSIONS: Our study is the first in vitro study assessing the impact of ADSCs on NP cells in an un-physiological mechanical stimulation culture environment. Our study noted that ADSCs protect compressive load induced NP cell death and degradation by inhibition of activated caspase-9 and -3 activity; regulating ECM and modulator genes, suppressing pro-inflammatory factors and preserving CK8. Consequently, the protective impact of ADSCs found in this study provides an essential understanding and expands our knowledge as to the utility of ADSCs therapy for intervertebral disc regeneration.published_or_final_versio

Immune cascades in human intervertebral disc: the pros and cons

The unique structural hallmark of the intervertebral disc has made its central composition, the nucleus pulposus (NP), excluded from the immunologic tolerance. Consequently, the intervertebral disc is identified as an immune-privileged organ. Traditionally, local detrimental immune activities caused by NP at the lesion sites of the disc are noted as a significant factor contributing to disc degeneration. However, given the beneficial activities of immune cells in other immune-privileged sites on basis of current evidence, the degenerate disc might need the assistance of a subpopulation of immune cells to restore its structure and lessen inflammation. In addition, the beneficial impact of immune cells can be seen in the absorption of the herniated NP, which is an important factor causes the mechanical compression of nerve roots. Consequently, a modulated immune network in degenerate disc is essential for the restoration of this immune-privileged organ. Until now, the understandings of immune response in disc degeneration still rest on the harmful aspect. Further studies are needed to explore its beneficial influence. Accordingly, there are no absolutely the pros and cons in terms of immune reactions caused by NP.published_or_final_versio

Magnetic field effects on the electroluminescence of organic light emitting devices: A tool to indicate the carrier mobility

The magnetoelectroluminescence (MEL) of organic light emitting devices with a N, N′ -bis(l-naphthyl)- N, N′ -diphenyl- 1, l′ -biphentl- 4, 4′ -diamine:tris-(8-hydroxyquinoline) aluminum (NPB: Alq 3) mixed emission layer (EML) has been investigated. We find that MEL is maximized when the volume ratio of NPB of the mixed EML reaches 30% and the EML thickness is 40 nm. The features of MEL under various magnetic field strengths are insensitive to the change in EML thickness and mixing ratio. Meanwhile, MEL has a close relationship with the carrier mobility. We have conducted a theoretical study to further verify the relationship. Our experimental and theoretical results confirm that MEL can function as a tool to indicate the mobility. © 2010 American Institute of Physics.published_or_final_versio

FasL expression on human nucleus pulposus cells contributes to the immune privilege of intervertebral disc by interacting with immunocytes

The mechanisms of immune privilege in human nucleus pulposus (NP) remain unclear. Accumulating evidence indicates that Fas ligand (FasL) might play an important role in the immune privilege of the disc. We aimed for addressing the role of FasL expression in human intervertebral disc degeneration (IDD) and immune privilege in terms of the interaction between NP cells and immunocytes via the FasL-Fas machinery. We collected NP specimens from 20 patients with IDD as degenerative group and 8 normal cadaveric donors as control. FasL expression was detected by qRT-PCR, western blotting and flow cytometry (FCM). We also collected macrophages and CD8(+) T cells from the peripheral blood of patients with IDD for co-cultures with NP cells. And macrophages and CD8(+) T cells were harvested for apoptosis analysis by FCM after 2 days of co-cultures. We found that FasL expression in mRNA, protein and cellular resolutions demonstrated a significant decrease in degenerative group compared with normal control (p<0.05). FCM analysis found that human NP cells with increased FasL expression resulted in significantly increased apoptosis ratio of macrophages and CD8(+) T cells. Our study demonstrated that FasL expression tends to decrease in degenerated discs and FasL plays an important role in human disc immune privilege, which might provide a novel target for the treatment strategies for IDD.published_or_final_versio

Down-regulated CK8 expression in human intervertebral disc degeneration

As an intermediate filament protein, cytokeratin 8 (CK8) exerts multiple cellular functions. Moreover, it has been identified as a marker of notochord cells, which play essential roles in human nucleus pulposus (NP). However, the distribution of CK8 positive cells in human NP and their relationship with intervertebral disc degeneration (IDD) have not been clarified until now. Here, we found the percentage of CK8 positive cells in IDD (25.7+/-4.14%) was significantly lower than that in normal and scoliosis NP (51.9+/-9.73% and 47.8+/-5.51%, respectively, p<0.05). Western blotting and qRT-PCR results confirmed the down-regulation of CK8 expression in IDD on both of protein and mRNA levels. Furthermore, approximately 37.4% of cell clusters were CK8 positive in IDD. Taken together, this is the first study to show a down-regulated CK8 expression and the percentage of CK8 positive cell clusters in IDD based upon multiple lines of evidence. Consequently, CK8 positive cells might be considered as a potential option in the development of cellular treatment strategies for NP repair.published_or_final_versio

H-alpha +[NII] Observations of the HII Regions in M81

In a first of a series of studies of the H-alpha + [NII] emission from nearby spiral galaxies, we present measurements of H-alpha + [NII] emission from HII regions in M81. Our method uses large-field-CCD images and long-slit spectra, and is part of the ongoing Beijing-Arizona-Taipei-Connecticut Sky Survey. The CCD images are taken with the NAOC 0.6/0.9m f/3 Schmidt telescope at the Xinglong Observing Station, using a multicolor filter set. Spectra of 10 of the brightest HII regions are obtained using the NAOC 2.16m telescope with a Tek 1024 X 1024 CCD. The continua of the spectra are calibrated by flux-calibrated images taken from the Schmidt observations. We determine the continuum component of our H-alpha + [NII] image via interpolation from the more accurately-measured backgrounds (M81 starlight) obtained from the two neighboring (in wavelength) BATC filter images. We use the calibrated fluxes of H-alpha + [NII] emission from the spectra to normalize this interpolated, continuum-subtracted H-alpha + [NII] image. We estimate the zero point uncertainty of the measured H-alpha + [NII] emission flux to be $\sim$ 8%. A catalogue of H-alpha + [NII] fluxes for 456 HII regions is provided, with those fluxes are on a more consistent linear scale than previously available. The logarithmically-binned H-alpha + [NII] luminosity function of HII regions is found to have slope $\alpha$ = -0.70, consistent with previous results (which allowed $\alpha=-0.5 \sim -0.8$). From the overall H-alpha + [NII] luminosity of the HII regions, the star formation rate of M81 is found to be $\sim 0.68 M_{\odot} {\rm yr}^{-1}$, modulo uncertainty with extinction corrections.Comment: 18 pages, 7 figures, accepted for publication in the Astronomical Journa

The Domestication Process and Domestication Rate in Rice: Spikelet Bases from the Lower Yangtze

The process of rice domestication occurred in the Lower Yangtze region of Zhejiang, China, between 6900 and 6600 years ago. Archaeobotanical evidence from the site of Tianluoshan shows that the proportion of nonshattering domesticated rice (Oryza sativa) spikelet bases increased over this period from 27% to 39%. Over the same period, rice remains increased from 8% to 24% of all plant remains, which suggests an increased consumption relative to wild gathered foods. In addition, an assemblage of annual grasses, sedges, and other herbaceous plants indicates the presence of arable weeds, typical of cultivated rice, that also increased over this period

Design, synthesis and biological characterization of novel inhibitors of CD38

Human CD38 is a novel multi-functional protein that acts not only as an antigen for B-lymphocyte activation, but also as an enzyme catalyzing the synthesis of a Ca 2+ messenger molecule, cyclic ADP-ribose, from NAD +. It is well established that this novel Ca 2+ signaling enzyme is responsible for regulating a wide range of physiological functions. Based on the crystal structure of the CD38/NAD + complex, we synthesized a series of simplified N-substituted nicotinamide derivatives (Compound1-14). A number of these compounds exhibited moderate inhibition of the NAD + utilizing activity of CD38, with Compound4 showing the highest potency. The crystal structure of CD38/Compound4 complex and computer simulation of Compound7 docking to CD38 show a significant role of the nicotinamide moiety and the distal aromatic group of the compounds for substrate recognition by the active site of CD38. Biologically, we showed that both Compounds4 and 7 effectively relaxed the agonist-induced contraction of muscle preparations from rats and guinea pigs. This study is a rational design of inhibitors for CD38 that exhibit important physiological effects, and can serve as a model for future drug development. © 2011 The Royal Society of Chemistry.postprin