Objective To summarize the research progress of platelet-rich plasma (PRP) for the treatment of discogenic low back pain (DLBP). Methods The literature on the treatment of DLBP with PRP was extensively reviewed, and the classification, treatment mechanism, in vitro and in vivo experiments and clinical trial progress of PRP were summarized. Results According to the PRP composition, preparation methods, and physicochemical properties, there are five commonly used PRP classification systems at present. PRP is involved in delaying or reversing the progress of disc degeneration and pain control by promoting the regeneration of nucleus pulposus cells, increasing the synthesis of extracellular matrix, and regulating the internal microenvironment of degenerative intervertebral disc. Although several in vitro and in vivo studies have confirmed that PRP can promote disc regeneration and repair, significantly relieve pain, and even improve the mobility of DLBP patients. But the contrary conclusion has been reached in a few studies, and there are limitations to the application of PRP. ConclusionCurrent studies have confirmed the effectiveness and safety of PRP in the treatment of DLBP and intervertebral disc degeneration, as well as the advantages of PRP in terms of ease of extraction and preparation, low immunological rejection, high regenerative and repair capacity, and the ability to compensate for the shortcomings of traditional treatment modalities. However, relevant studies are still needed to further optimize PRP preparation methods, unify systematic classification guidelines, and clarify its long-term effectiveness.
ObjectiveTo analyze the relationship between the bone mineral density (BMD) and lumbar intervertebral disc degeneration in rhesus macaques by using T1ρ-MRI.
MethodsTwenty female rhesus macaques at the age of 10.9 years on average (rang, 4-20 years) were selected. The lumbar intervertebral discs were classified by Pfirrmann grading system and the T1ρ relaxation time (T1ρ value) was examined by using MRI (Philips 1.5 Tesla), and then BMD values of the L4,5 vertebrae and femoral ward's triangle were detected by using Osteocore dual energy X-ray absorptiometry. Finally, the relationship of T1ρ value of the lumbar intervertebral discs and Pfirrmann grading with age, weight, BMD of lumbar vertebrae and femoral ward's triangle was analyzed.
ResultsThe BMD values of lumbar vertebrae and femoral ward's triangle were (0.64±0.17) g/cm2 and (0.67±0.19) g/cm2 respectively, showing no significant difference (t=2.893, P=0.128). According to Pfirrmann grading system, there were 7 cases of grade I, 8 cases of grade Ⅱ, and 5 cases of grade Ⅲ at L4,5 intervertebral discs. The T1ρ value of the lumbar intervertebral disc was (104.08±18.65) ms; the T1ρ values of grades I, Ⅱ, and Ⅲ were (121.31±13.44), (104.73±15.01), and (77.41±11.87) ms, respectively. There was a negative correlation between T1ρ value and the age and the BMD of lumbar vertebrae and femoral ward's triangle. There was a positive correlation between Pfirrmann grading and the variables as listed above. Significant negative linear correlation was also observed between T1ρ value and Pfirrmann grading.
ConclusionThe T1ρ value is a reliable index when quantifying lumbar intervertebral disc degeneration, and there is a significant positive correlation between BMD and lumbar intervertebral disc degeneration in rhesus macaques.
ObjectiveTo investigate the influence of endoplasmic reticulum stress (ERS) on smoking-induced nucleus pulposus cells apoptosis and inflammatory response.MethodsBetween October 2016 and October 2018, 25 patients with cervical disc herniation receiving discectomy were collected and divided into smoking group (14 cases) and non-smoking group (11 cases). The baseline data of age, gender, herniated segment, and Pfirrmann grading showed no significant difference between the two groups (P>0.05). The obtained nucelus pulposus tissues were harvested to observe the cell apoptosis via detecting the apoptosis-related proteins (Caspase-3 and PRAP) by TUNEL staining and Western blot test. The nucleus pulposus cells were isolated and cultured with enzyme digestion, of which the third generation cells were used in follow-up experiments. Then, the expressions of inflammatory factors [interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α)] were detected by ELISA; the nuclear translocation of P65 was monitored by cell immunofluorescence staining. Furthermore, ERS-related proteins (GRP78 and CHOP) were detected by Western blot; and endoplasmic reticulum ultrastructure was observed under transmission electron microscope. To verify the regulatory effect of ERS, cells were pretreated by ERS specific inhibitor (4-PBA), then cell apoptosis and inflammatory response were tested.ResultsThe nucleus pulposus tissue observation showed that the cell apoptotic rate and the expressions of apoptosis-related proteins (Caspase-3 and PARP) were obviously higher in smoking group than in non-smoking group (P<0.05). The nucleus pulposus cells observation indicated that the expressions of the inflammatory factors (IL-1β and TNF-α) and the ERS-related proteins (GRP78 and CHOP) were also higher in smoking group than in non-smoking group (P<0.05). The results of cell immunofluorescence staining further confirmed that smoking stimulated nuclear translocation of P65 in nucleus pulposus cells. The ERS injury was much more serious in smoking group than in non-smoking group. Furthermore, after 4-PBA inhibiting ERS, the expressions of GRP78, CHOP, IL-1β, TNF-α, and P65 were significantly decreased (P<0.05), and flow cytometry results showed that cell apoptotic rate in smoking group was decreased, showing significant difference compared with the non-smoking group (P<0.05).ConclusionSomking can stimulate cell apoptosis and inflammatory response in nucleus pulposus cells via ESR pathway. Suppressing ESR may be a novel target to suspend smoking-induced intervertebral disc degeneration.
Objective To evaluate the effectiveness and safety of different injection sites for collagenase chemonucleolysis for lumbar intervertebral disc protrusion (LIDP). Methods We searched for randomized controlled trials (RCTs) and quasi-RCTs in the following electronic databases: Pubmed (1966 to May 2006), EMBASE(1966 to May 2006), The Cochrane library (Issue 2, 2006), CRD(Center for Reviews and Dissemination),York University, CBM (1978 to May 2006 ), CNKI(1994-2006)and VIP(1989-2006). Quality assessment and data extraction were conducted by two reviewers independently. Disagreement was resolved through discussion. Results Eight studies involving a total of 1035 participants met the inclusion criteria. Meta-analysis was not carried out because of apparent heterogeneity. Four studies made comparisons among intradisc, extradisc, and combined intra- and extra-disc injection. One study (62 participants) showed that intradisc injection was superior to extradisc injection (RR 3.71, 95% CI 1.19 to 11.58, Plt;0.05). Another study (240 participants) showed that intradisc injection was superior to combined intra- and extra-disc injection after 3 months and 6 months of follow-up (RR 0.88, 95% CI 0.80 to 0.98, Plt;0.05). The other two studies showed no significant difference among intradisc, extradisc, and combined intra- and extra-disc injection. Four studies (436 participants in total) showed that amongst three extradisc injections, both anterior epidural space injection and lateral epidural space injection were superior to posterior epidural space injection (Plt;0.05). Although one study indicated that anterior epidural space injection was superior to lateral epidural space injection (RR 1.24, 95% CI 1.03 to 1.51, Plt;0.05), no statistical significance was found between anterior epidural space injection and lateral epidural space injection in two other studies (Pgt;0.05). Conclusions There is insufficient evidence to identify which injection site for collagenase is the most effective for lumbar intervertebral disc protrusion. The included studies showed that both anterior and lateral epidural space injection were superior to posterior epidural space injection. However, these studies are too small and poor quality, and have different diagnostic criteria, follow-up time points, outcome measures and efficacy parameters. Thus, more high quality and large-scale RCTs are needed.
ObjectiveTo explore the feasibility and effectiveness of unilateral pedicle screw rod and single poly (ether-ether-ketone) (PEEK) Cage for lumbar intervertebral disc protrusion.
MethodsA total of 126 cases of single segment of lumbar intervertebral disc protrusion between January 2006 and June 2012 were divided into 2 groups in the randomized clinical trial. Unilateral pedicle screw fixation and single PEEK Cage was used in 63 cases (research group), and bilateral pedicle screw fixation and single PEEK Cage in 63 cases (control group). There was no significant difference in gender, age, disease duration, side, and affected segment between 2 groups (P>0.05). Schulte evaluation criterion was used to assess bone graft fusion, Oswestry disability index (ODI) to estimate the quality of life situation, and visual analogue scale (VAS) to evaluate the improvements of lower back pain. Macnab standards was applied to assess postoperative effectiveness, and Emery ways to measure the height of intervertebral space.
ResultsThe incision length, operation time, intraoperative blood loss, hospitalization time, and hospitalization fee in research group were significantly less than those in control group (P<0.05). The patients were followed up 12-79 months (mean, 21.3 months) in research group, and 15-73 months (mean, 22.6 months) in control group. The postoperative lordosis was recovered well, and the height of intervertebral space was increased. No loosening or breakage of internal fixation occurred. The time of bone graft fusion was (6.8±1.3) months in research group and was (7.1±1.2) months in control group, showing no significant difference (t=1.153, P=0.110). The height of intervertebral space, ODI score, and VAS score were significantly improved when compared with preoperative ones in 2 groups (P<0.05), but no significant difference was found between 2 groups at preoperation and last follow-up (P>0.05). At 3 months after operation, postoperative effectiveness was assessed according to Macnab criterion, the excellent and good rate was 95.23% in research group (excellent in 13 cases, good in 47 cases, and fair in 3 cases) and was 71.42% in control group (excellent in 7 cases, good in 38 cases, fair in 15 cases, and poor in 3 cases); the research group was significantly superior to control group (χ2=6.110, P=0.006).
ConclusionUnilateral pedicle screw fixation and single PEEK Cage has the advantages of small trauma, reliable fixation, shorter operation time, less bleeding, less economic cost, and early off-bed activity time. It can provide a simple and reliable choice in single segmental lumbar intervertebral disc protrusion.
Objective
To fabricate a novel composite scaffold with acellular demineralized bone matrix/acellular nucleus pulposus matrix and to verify the feasibility of using it as a scaffold for intervertebral disc tissue engineering through detecting physical and chemical properties.
Methods
Pig proximal femoral cancellous bone rings (10 mm in external diameter, 5 mm in internal diameter, and 3 mm in thickness) were fabricated, and were dealed with degreasing, decalcification, and decellularization to prepare the annulus fibrosus phase of scaffold. Nucleus pulposus was taken from pig tails, decellularized with Triton X-100 and deoxycholic acid, crushed and centrifugalized to prepare nucleus pulposus extracellular mtrtix which was injected into the center of annulus fibrosus phase. Then the composite scaffold was freeze-dryed, cross-linked with ultraviolet radiation/carbodiimide and disinfected for use. The scaffold was investigated by general observation, HE staining, and scanning electron microscopy, as well as porosity measurement, water absorption rate, and compressive elastic modulus. Adipose-derived stem cells (ADSCs) were cultured with different concentrations of scaffold extract (25%, 50%, and 100%) to assess cytotoxicity of the scaffold. The cell viability of ADSCs seeded on the scaffold was detected by Live/Dead staining.
Results
The scaffold was white by general observation. The HE staining revealed that there was no cell fragments on the scaffold, and the dye homogeneously distributed. The scanning electron microscopy showed that the pore of the annulus fibrosus phase interconnected and the pore size was uniform; acellular nucleus pulposus matrix microfilament interconnected forming a uniform network structure, and the junction of the scaffold was closely connected. The novel porous scaffold had a good pore interconnectivity with (343.00 ± 88.25) μm pore diameter of the annulus fibrosus phase, 82.98% ± 7.02% porosity and 621.53% ± 53.31% water absorption rate. The biomechanical test showed that the compressive modulus of elasticity was (89.07 ± 8.73) kPa. The MTT test indicated that scaffold extract had no influence on cell proliferation. Live/Dead staining showed that ADSCs had a good proliferation on the scaffold and there was no dead cell.
Conclusion
Novel composite scaffold made of acellular demineralized bone matrix/acellular nucleus pulposus matrix has good pore diameter and porosity, biomechanical properties close to natural intervertebral disc, non-toxicity, and good biocompatibility, so it is a suitable scaffold for intervertebral disc tissue engineering.
Objective To explore the osteogenic potential of cervical intervertebral disc fibroblasts in vitro, to investigate the regulatory factors of recombinant human bone morphogenetic protein 2(rhBMP-2) and tumor necrosis factor α(TNF-α) on osteogenic phenotype of fibroblasts and to discuss the condition that facilitates osteogenesis of fibroblasts. Methods Theannulus fibroblasts cell lines of experiment goats were established in vitro and the biologicspecificity was found. According to different medias, 4 groups were included in this experiment: control group, TNF-α group ( 50 U/ml TNF-α), rhBMP-2 group (0.1 μg/ml rhBMP-2) and TNF-α+rhBMP-2 group (50 U/ml TNF-α+0.1 μg/ml rhBMP-2). Thefibroblasts were incubated in the media for about 3 weeks,and then the markers for osteogenic features were investigated by biochemistry, histochemistry observations. Results rhBMP-2 and TNF-α had no effect on the proliferation of fibroblasts from the experiment goats. rhBMP-2 or TNF-α could stimulate fibroblasts to secrete alkaline phosphatase and collagen type Ⅰ. The combined use of rhBMP-2 and TNF-α or the single use of rhBMP-2 could make fibroblasts to secrete osteocalin and the morphological changes of the fibroblasts were very obvious. Histochemical study of the nodules with specific new bone labeler(Alizarin red S) revealed positive reaction, denoting that the nodules produced by the fibroblasts werebone tissues. There was statistically significant difference(Plt;0.05) inALP activity between 3 experimental groups and control group and in secretion of osteocalcin between rhBMP-2 group, TNF-α+rhBMP-2 group and control group. Conclusion The results point out clearly that rhBMP-2 can induce theosteogenic potential of annulus fibroblasts in vitro.
Objective
To develop a modified short time inversion recovery (STIR) sequence grading system for lumbar intervertebral disc degeneration based on MRI STIR sequences, and to test the validity and reproducibility of this grading system.
Methods
A modified 8-level grading system for lumbar intervertebral disc degeneration based on routine sagittal STIR sequences and modified Pfirrmann grading system was developed. Between April 2011 and February 2012, 60 patients with different degrees of lumbar intervertebral disc degeneration were selected as objects of study, including 32 males and 28 females with an average of 50 years (range, 17-85 years). T2 weighted and STIR sequence images were obtained from the lumbar discs of L1, 2-L5, S1 of each object (total, 300 discs). All examinations were analyzed independently by 3 observers and a consensus readout was performed after all data collected. The validity and reproducibility were analyzed by calculating consistent rate and Kappa value.
Results
According to the grading system, there were 0 grade 1, 83 (27.7%) grade 2, 87 (29.0%) grade 3, 66 (22.0%) grade 4, 31 (10.3%) grade 5, 15 (5.0%) grade 6, 12 (4.0%) grade 7, and 6 (2.0%) grade 8. Intra-observer consistency was b (Kappa value range, 0.822-0.952), and inter-observer consistency was high to b (Kappa value range, 0.749-0.843). According to the consensus analysis, the total consistent rate was 82.7%-92.7% (mean, 85.6%). A difference of one grade occurred in 13.9% and a difference of two or more grades in 0.5% of all the cases.
Conclusion Disc degeneration can be graded by using modified STIR sequence grading system, which can improve the accuracy of grading different degrees of lumbar intervertebral disc degeneration.
The lumbar intervertebral disc exhibits a complex physiological structure with interactions between various segments, and its components are extremely complex. The material properties of different components in the lumbar intervertebral disc, especially the water content (undergoing dynamic change as influenced by age, degeneration, mechanical loading, and proteoglycan content) - critically determine its mechanical properties. When the lumbar intervertebral disc is under continuous pressure, water seeps out, and after the pressure is removed, water re-infiltrates. This dynamic fluid exchange process directly affects the mechanical properties of the lumbar intervertebral disc, while previous isotropic modeling methods have been unable to accurately reflect such solid-liquid phase behaviors. To explore the load-bearing mechanism of the lumbar intervertebral disc and establish a more realistic mechanical model of the lumbar intervertebral disc, this study developed a solid-liquid biphasic, fiber-reinforced finite element model. This model was used to simulate the four movements of the human lumbar spine in daily life, namely flexion, extension, axial rotation, and lateral bending. The fluid pressure, effective solid stress, and liquid pressure-bearing ratio of the annulus fibrosus and nucleus pulposus of different lumbar intervertebral discs were compared and analyzed under the movements. Under all the movements, the fluid pressure distribution was closer to the nucleus pulposus, while the effective solid stress distribution was more concentrated in the outer annulus fibrosus. In terms of fluid pressure, the maximum fluid pressure of the lumbar intervertebral disc during lateral bending was 1.95 MPa, significantly higher than the maximum fluid pressure under other movements. Meanwhile, the maximum effective solid stress of the lumbar intervertebral disc during flexion was 2.43 MPa, markedly higher than the maximum effective solid stress under other movements. Overall, the liquid pressure-bearing ratio under axial rotation was smaller than that under other movements. Based on the solid-liquid biphasic modeling method, this study more accurately revealed the dominant role of the liquid phase in the daily load-bearing process of the lumbar intervertebral disc and the solid-phase mechanical mechanism of the annulus fibrosus load-bearing, and more effectively predicted the solid-liquid phase co-load-bearing mechanism of the lumbar intervertebral disc in daily life.
Objective To investigate the feasibility oftissue engineered intervertebral disc for regeneration of discs. Methods A three-dimensional porous poly(L-lactic-co-glycolic acid) (PLGA) scaffold was fabricated by temperature induced phase separation method. Human fetal disc cells were isolated and cultured in vitro. The disc cells labeledwith a PKH-26 fluorescent dye were seeded into a threedimensional porous scaffold. The proliferation of disc cells with PKH-26 fluorescent labels was assessed by using MTT uptake, laser fluorescence microscopy and SEM. Results Human fetal disc cells displayed a polygonal shape in primary monolayer culture. A regular arrangement and microtubules orientationstructure scaffold with 50-300 μm in diameter was fabricated by thermal-induced phase separation technique. MTT uptake and fluorescent microscopy examination indicated that the seeded disc cells were viable and showed proliferation activity within a porous scaffold. Conclusion The above findings support potential applications of tissue engineered disc in treatment of disc degenerative diseases.
