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.
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.
To detect the cell density, apoptotic incidence and the expressions of Bax and Caspase-3in human lumbar intervertebral discs, so as to further understand the mechanism of human lumbar intervertebral discdegeneration and provide a new idea for biologic treatment of it in future. Methods From May to December in 2006,30 human lumbar intervertebral discs in experimental group(L2 to S1)were surgically collected from 27 patients undergoing posterior lumbar intervertebral discoidectomy and fusion. All the cases were affirmed by MRI and they never experienced discography, collagenolysis of nucleus pulposus and percutaneous laser disc decompression. The control group consisted of 20 human lumbar intervertebral discs(L2 to S1)harvested from 5 young men without spine-related condition immediately after their accidental death. Apoptotic disc cells were detected by TUNEL and histomorphology, and immunohistochemical staining with SP method was performed to examine the expressions of Bax and Caspase-3 in all specimens. Results HE staining disclosed that the average cell density in control group (17.16 ± 1.22)/HP was higher than that in experimental group (12.41 ± 0.95)/HP (P lt; 0.01). However, TUNEL staining observed that the average TUNEL positive incidence in control group (6.97% ± 0.92%) was lower than that in experimental group (12.59% ± 0.95%), (P lt; 0.01). Immunohistochemical staining with SP method showed that the Bax and Caspase-3 positive incidence of nucleus pulposus in control group (11.02% ± 1.18%, 9.01% ± 1.00%) were lower than those in experimental group (19.29% ± 1.18%, 15.07% ± 0.97%), (P lt; 0.01). The results of the average gray scale value of nucleus pulposus in control group were 187.33 ± 7.88 and 185.68 ± 3.26, respectively, with 124.98 ±6.69 and 160.13 ± 4.37 in experimental group. There was significant difference between the two groups (P lt; 0.01). When thetotal 50 specimens in the two groups were analyzed, TUNEL positive incidence showed significant inverse correlations with their respectively corresponding cell densities (r = － 0.88, r = － 0.93, P lt; 0.01). The Bax and Caspase-3 positive incidence of nucleus pulposus showed significant positive correlation with the TUNEL positive incidence of nucleus pulposus (r = 0.83, r = 0.91, P lt; 0.01). Conclusion The decrease of cell density is involved in the development of human lumbar intervertebral disc degeneration. Bax and Caspase-3 might play a role in disc cell apoptosis in nucleus pulposus of human lumbar intervertebral disc.
ObjectiveTo discuss the safety and effectiveness of combined dynamic cervical implant (DCI) and Cage fusion in the treatment of two-level cervical disc protrusion. MethodsBetween September 2009 and June 2011, 16 cases of two-level cervical disc protrusion were treated with combined DCI and Cage fusion. Of 16 cases, 10 were male and 6 were female, with a mean age of 44.1 years (range, 37-64 years) and with a mean disease duration of 5.1 years (range, 2-8 years), including 8 cases of cervical myelopathy, 5 cases of nerve root cervical myelopathy, and 3 cases of mixed cervical myelopathy. Radiological results indicated degenerative intervertebral discs and compressed never root or spinal cord. Involved discs included C3,4 and C4,5 (1 case), C3,4 and C5,6(5 cases), C4,5 and C5,6 (3 cases), C4,5 and C6,7 (5 cases), and C5,6 and C6,7 (2 cases). The neck disability index (NDI), Japanese Orthopedic Association (JOA) score, and visual analogue scale (VAS) were used to evaluate the neurological function and pain relief. The stabilities and activities of involved segments, intervertebral fusion, and displacement of Cages were observed during follow-up. ResultsPrimary healing of incisions was obtained in all cases; no complication of hematoma, infection, cerebrospinal fluid leakage, or neural function damage occurred. All 16 patients were followed up 18 months on average (range, 6-36 months). The cervical X-ray results indicated that the activities of involved segments was (7.8±3.1)°, showing no significant difference (t=0.655, P=0.132) when compared with preoperative value [(7.3±2.6)°]. No implant loosening was observed; slight heterotopic ossification occurred in 1 patient at the posterior rim of intervertebral space. No cage loosening or sinking was seen, and good fusion was achieved. The mean time of fusion was 4.5 months (range, 3-8 months). NDI, JOA, and VAS scores at last follow-up (18.3±5.1, 15.7±1.5, and 3.4±1.8 respectively) were significantly improved (t=2.131, P=0.016; t=3.126, P=0.024; t=6.102, P=0.038) when compared with preoperative scores (49.6±11.3, 12.8±2.0, and 6.7±1.2 respectively). ConclusionA combination of DCI and intervertebral Cage fusion has satisfactory early effectiveness in treatment of two-level cervical intervertebral protrusion for maintaining the stability and activity of cervical vertebrae.
Objective Platelet-rich plasma (PRP) can stimulate intervertebral disc cell proliferation, promote extracellular matrix synthesis, and inhibit annulus fibrosus cell apoptosis. To investigate the effects of autologous PRP on the treatment of the early intervertebral disc degeneration (IDD) so as to provide the experimental basis for its clinical application. Methods Forty-five healthy New Zealand white rabbits (male or female, weighing 2.5-3.0 kg) were randomly divided into the experimental group (n=15), the control group (n=15), and the sham group (n=15). PRP was prepared from the arterial blood of rabbit’s ears of the experimental group with Landesberg’s method. The platelet concentrations in both whole blood and PRP were detected. The rabbit model of early IDD was established by annulus fibrosus puncture (L4, 5, L5, 6) in both the experimental group and the control group; 100 ?L autologous PRP and 100 ?L PBS were injected into the degenerative intervertebral discs respectively after 2 weeks of models creation. In sham group, intervertebral discs were separated and exposed without treatment. The general conditions of the rabbits were observed after building models; at 2 weeks after degeneration, 1 and 2 weeks after intervention, 5 rabbits were selected randomly from each group respectively for MRI observation, histological observation by using HE staining and collagen type II immunohistochemical staining. The signal of lumbar MRI was assessed and the contents of collagen type II were detected. Results The platelet concentration of PRP was about 4.92 times as much as that of the whole blood. All the animals survived to the end of the experiment. At 2 weeks after degeneration, a lower T2 signal was observed in both the experimental group and the control group; the nucleus pulposus cells decreased and extracellular matrix degenerated; and the expression of collagen type II decreased in both the experimental group and control group. The degenerative grade of lumbar MRI in the experimental group and control group were significantly higher than that in the sham group (P lt; 0.05), and the content of collagen type II were significantly lower than that in the sham group (P lt; 0.05). At 1, 2 weeks after intervention, disc degeneration in the experimental group was significantly lower than that in control group (P lt; 0.05), and significant difference was found between experimental group and sham group (P lt; 0.05). The nucleus pulposus cells and chondroid matrix in the experimental group were more than those in the control group, showing slight stromal fibrosis; but the expression of collage type II was significantly higher than that in the control group (P lt; 0.05). Conclusion The disc injection of autologous PRP may terminate or even reverse the progress of rabbit early IDD, which may be associated with the role of multiple growth factors of PRP in regulating cell function, improving the tissue microenvironment, and promoting tissue regeneration.
OBJECTIVE To observe the effects of sodium hyaluronate on nerve root adhesion after operation of lumbar disc protrusion (LDP). METHODS From April 1996 to June 2001, 152 cases with LDP were performed the removal of nucleus pulposus under endoscope. After operation, 2 ml of sodium hyaluronate were injected. The clinical results were evaluated by clinical symptom, straight leg raising test, patient’s satisfaction degree and back pain. RESULTS All the patients were followed for 1 month to 3 years. The average angle of straight leg raising test increased from 26 degree preoperatively to 62 degree postoperatively. The patient’s pain were markedly relieved. Patient’s satisfactory degree were 86.8%. CONCLUSION The injection of sodium hyaluronate after operation of LDP has effect on preventing postoperative nerve root adhesion.
Objective To evaluate the effect and safety of the conservative treatment for the protrusion of cervical vertebra intervertebral disc. Methods We searched CBM disk (1978~2007), CNKI (1979~2007), VIP (1989~2007). Telephone interviewed the original authors of claimed randomized controlled trials(RCTs) to identify whether they are truly RCTs. Only true RCTs and quasi-randomized controlled trials were included and the quality of them was critically assessed. Results Five RCTs and eleven quasi-randomized controlled trials were identified and included. It showed that lifting-massage, massage combining with traction or acupuncture, articulus mobilization combining with traction and electrotherapy, traction combining with computer intermediate frequency therapeusis and injectio ad acumen combining with massage have marked improvement. But it could hardly draw a conclusion that all the combining therapies had better curative effect. Conclusions Weak evidence shows that combining conservative treatment can improve the curative effect of the protrusion of cervical vertebra intervertebral disc. The results need to be supported by well designed and high quality trials.
ObjectiveTo review the research progress of the role and mechanism of adipokines in intervertebral disc degeneration (IVDD) in recent years.MethodsThe domestic and foreign literature related to adipokines in the process of IVDD was extensively reviewed. The types and functions of adipokines, the role and mechanism in the process of IVDD, and the application prospects of intervertebral disc biotherapy were reviewed.ResultsAs a kind of bioactive substance secreted by adipose tissue, adipokine plays an important role in bone and joint diseases, metabolic diseases, and breast cancer. During IVDD, most adipokines can activate multiple signaling pathways by binding to autoreceptors, cause the proliferation and apoptosis of cells and proinflammatory and anti-inflammatory factors parasecretions in the intervertebral disc, and lead to imbalance of intradiscal metabolism and establishment of the initial inflammatory environment, and finally cause the IVDD.ConclusionAdipokines, as a biologically active substance with metabolic and immunomodulatory functions, play important roles in the occurrence, development, and biological treatment of IVDD.
摘要:目的:探讨联合应用激光汽化减压(percutaneous laser disc discompression,PLDD)、射频热凝靶点消融、臭氧注射治疗腰椎间盘突出症的的个体化选择。方法: 自2006年6月,在CT引导下选择性联合应用PLDD、射频和臭氧治疗腰椎间盘突出症患者267例,突出椎间盘的特点个体化选择穿刺路径和治疗方法;其中PLDD联合臭氧治疗92例(A组),射频联合臭氧治疗67例(B组),PLDD、射频和臭氧三者联合治疗108例(C组)。结果:所有患者均顺利完成手术,于术后1周、1个月,3个月及6个月随访记录VAS评分和Macanab优良率。三组患者VAS评分经方差分析,手术前、后有显著性差异(Plt;0.05),术后1周至6个月的VAS评分统计无显著性差异(Pgt;0.05);术后三组间VAS评分、Macanab优良率比较无显著性差异(Pgt;0.05)。结论: 选择性联合应用微创技术进行个体化的立体治疗,具有扩大微创手术适应症、提高手术疗效的优势,值得推广和利用。Abstract: Objective: To investigate the selectivity and individualization of using percutaneous laser disc discompression(PLDD) and ozone injection combined with radiofrequency thermocoagulation and target ablation curing lumbar intervertebral disc protrusion. Methods: From June 2006, 267 lumbar disc herniation cases were operated that guided by CT, the characteristic of the liable disc was confirmed by magnetic resonance imaging and CT before the procedure. 92 cases (A group) were treated by PLDD combined with ozone injection,67 case were treated by radiofrequency thermocoagulation and target ablation combined with ozone injection, 108 cases were treated by PLDD and ozone injection combined with radiofrequency thermocoagulation and target ablation. Results: All case been successfully operated, the theraptic effect was evaluated by comparing the value of VAS and excellent and good rate of therapy at preoperation and at 1 week, 1month,3 months, 6 months after operation. The value of VAS in three groups at postoperation were remarkably lower than preoperation (Plt;0.05). The excellent and good rate of therapy at 6 months was respectively 94.5％ in group A,94.0％ in group B and 95.4％ in group C,no significant difference was observed between the three groups(Pgt;0.05).Conclusion: The selectivity and individualization of using PLDD and ozone injection combined with radiofrequency thermocoagulation and target ablation curing lumbar intervertebral disc protrusion can enlarge the indication and improve the clinical curative effect, it should be spreaded in clinic.
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.