Extraction of RNA from tough tissues with high proteoglycan content by cryosection, second phase separation and high salt precipitation

published_or_final_versio

Hif-αs fine-tune Sox9-dependent Extracellular Matrix Production in Chondrocytes

INTRODUCTION: An avascular milieu is a hallmark feature of articular cartilage and therefore the expression of chondrocyte-specific matrix protein is orchestrated under hypoxic condition. Physiological responses under hypoxia are regulated by the ...postprin

The effect of severity of disc degeneration on mesenchymal stem cells' ability to regenerate the intervertebral disc: a rabbit model

Conference Theme: Spinal Motion Segment: From Basic Science to Clinical ApplicationThis Open Access journal suppl. entitled: Abstracts of European Cells and Materials VI /SRN I - Spinal Motion Segment: From Basic Science to Clinical Applicationpostprin

The effect of mesenchymal stem cells in stimulating intervertebral disc cells: a comparative study towards disc regeneration

Local Scholarship Awardees - Poster Sessions: no. 8DMM 2011 entitled: Re-engineering Regenerative MedicineINTRODUCTION: Intervertebral disc (IVD) degeneration is suggested to begin from the nucleus pulposus (NP). Evidence from various studies highlights mesenchymal stem cells (MSC), in most cases using bone marrow derived MSC, as a potential stem cell source for NP regeneration. However there are indications that fetal or close to fetal tissue sources contain MSC with relatively undifferentiated phenotype with respect to MSC from adult sources. Moreover, umbilical cord (C)-MSC may have better chondrogenic differentiation potential than bone marrow (B)-MSC. We hypothesize CMSC are more efficient than BMSC in stimulating NP ...postprin

A histological grading system for evaluating disc degeneration in the mouse

Poster no. 1634INTRODUCTION: Previously we have created a simple mouse model to study the disc degeneration in mice, using the technique of annular puncture in the mouse tail. The punctured mouse disc undergoes the progressive morphological changes over a period of 12 weeks. Degenerative changes were observed in both annulus fibrosus (AF) and nucleus pulposus (NP), such as appearance of cell clusters in NP and formation of cracks in AF. The objective of the current study is to establish a histological grading system to describe these degenerative changes and evaluate the severity of the degeneration in the punctured murine discs. We hope that this histological grading system can be useful in future study to evaluate the disc degeneration in the mouse …postprin

Tissue Engineering for Intervertebral Disk Degeneration

Many challenges confront intervertebral disk engineering owing to complexity and the presence of extraordinary stresses. Rebuilding a disk of native function could be useful for removal of the symptoms and correction of altered spine kinematics. Improvement in understanding of disk properties and techniques for disk engineering brings promise to the fabrication of a functional motion segment for the treatment of disk degeneration. Increasing sophistication of techniques available in biomedical sciences will bring its application into clinics. This review provides an account of current progress and challenges of intervertebral disk bioengineering and discusses means to move forward and toward bedside translation. © 2011.postprin

Intrinsic properties of mesemchymal stem cells from human bone marrow, umbilical cord and umbilical cord blood comparing the different sources of MSC

The past decade has witnessed numerous publications on mesenchymal stem cells (MSC), which have great potential in regenerative medicine. MSC from various types of origins exhibit different characteristics, which may relate to the maintenance role of MSC in that specific source. Reports have emerged that among the most widely investigated sources, umbilical cord (UC) or umbilical cord blood (UCB) derived MSC throw advantages over bone marrow (BM) derived MSC due to their close to fetal origin. Here the methodologies used to separate MSC from UC or UCB, and the intrinsic properties, including proliferation capacity, multipotency, cytokine profile, cell surface protein expression and gene expression, between UC, UCB and BM derived MSC, are discussed in details, though may not in a full picture, for the first time.postprin

Lumbar intervertebral disc allograft transplantation: long-term mobility and impact on the adjacent segments

Purpose: Fresh-frozen intervertebral disc (IVD) allograft transplantation has been successfully performed in the human cervical spine. Whether this non-fusion technology could truly decrease adjacent segment disease is still unknown. This study evaluated the long-term mobility of the IVD-transplanted segment and the impact on the adjacent spinal segments in a goat model. Methods: Twelve goats were used. IVD allograft transplantation was performed at lumbar L4/L5 in 5 goats; the other 7 goats were used as the untreated control (5) and for the supply of allografts (2). Post-operation lateral radiographs of the lumbar spine in the neutral, full-flexion and full-extension positions were taken at 1, 3, 6, 9 and 12 months. Disc height (DH) of the allograft and the adjacent levels was calculated and range of motion (ROM) was measured using the Cobb’s method. The anatomy of the adjacent discs was observed histologically. Results: DH of the transplanted segment was decreased significantly after 3 months but no further reduction was recorded until the final follow-up. No obvious alteration was seen in the ROM of the transplanted segment at different time points with the ROM at 12 months being comparable to that of the untreated control. The DH and ROM in the adjacent segments were well maintained during the whole observation period. At post-operative 12 months, the ROM of the adjacent levels was similar to that of the untreated control and the anatomical morphology was well preserved. Conclusions: Lumbar IVD allograft transplantation in goats could restore the segmental mobility and did not negatively affect the adjacent segments after 12 months.postprin

The paradoxical relationship between ligamentum flavum hypertrophy and developmental lumbar spinal stenosis

published_or_final_versio

Coupling of small leucine-rich proteoglycans to hypoxic survival of a progenitor cell-like subpopulation in Rhesus Macaque intervertebral disc

Degeneration of the intervertebral disc (IVD) is a major spinal disorder that associates with neck and back pain. Recent studies of clinical samples and animal models for IVD degeneration have identified cells with multi-potency in the IVD. However, IVD tissue-specific progenitor cells and their niche components are not clear, although degenerated IVD-derived cells possess invitro characteristics of mesenchymal stromal cell (MSCs). Here, we firstly identified the tissue-specific intervertebral disc progenitor cells (DPCs) from healthy Rhesus monkey and report the niche components modulated the survival of DPCs under hypoxia. DPCs possess clonogenicity, multipotency and retain differentiation potential after extended expansion invitro and invivo. In particular, the nucleus pulposus-derived DPCs are sensitive to low oxygen tension and undergo apoptosis under hypoxic conditions due to their inability to induce/stabilize hypoxia-inducible factors (HIF). The presence of small leucine-rich proteoglycans (SLRP), biglycan or decorin, can reduce the susceptibility of DPCs to hypoxia-induced apoptosis via promoting the activation/stabilization of HIF-1α and HIF-2α. As IVD is avascular, we propose SLRPs are niche components of DPCs in IVD homeostasis, providing new insights in progenitor cell biology and niche factors under a hypoxic microenvironment. © 2013 .postprin