ObjectiveTo review the mechanism and research progress of signal ing pathways which play key roles in the regulation of osteoblast differentiation and bone formation. MethodsRecent articles about signal ing pathways of osteoblast differentiation and bone formation were reviewed and comprehensively analyzed. ResultsAt present, multi ple signaling pathways have been found to be involved in the regulation of osteoblast differentiation and bone formation, among which bone morphogenetic protein-Smads, Wnt/β-catenin, Notch, Hedgehog, and fibroblast growth factor signaling pathways may play the most important roles. Not only each pathway has a complex regulatory mechanism itself, but also contacts and impacts with each other, thus they formed a more compl icated and sophisticated regulatory network, and regulate together osteoblast differentiation and bone formation. However, the mechanisms in detail of those pathways are still not very clear, because the animal experiment techniques are not yet mature as well as the relevant cl inical trials were carried out not too much. ConclusionThe complete molecular mechanism of osteoblast differentiation and bone formation should be further investigated, so as to lay a theory foundation for preventing and treating the common bone diseases in cl inical which are involve in osteoblast differentiation and bone formation.
ObjectiveTo review the biomaterial and clinical prospects of annulus fibrosus tissue engineering. MethodsThe recent literature concerning annulus fibrosus tissue engineering, including cell source, bioactive molecules, and biomaterial was extensively reviewed and summarized. ResultsMesenchymal stem cells (MSCs) is an ideal seed cells. When annulus fibrosus cells and MSCs in the ratio of 2:1 are cultured, it shows the closest mRNA expression levels of annulus fibrosus-related markers. Bioactive molecules can be divided into 4 types:growth factors, morphogens, catabolic enzyme inhibitors, and intracellular regulators. They play an active role in promoting the synthesis of extracellular matrix, and maintaining intervertebral disc homeostasis and a balance between anabolic- and catabolic process in the disc. Based on the source, biological materials can be divided into natural materials, synthetic materials, and composite materials. The mechanical properties of the annulus fibrosus is an important basis for material design. Up to now, none of these scaffold materials is accepted as the most suitable one. The selection of scaffold materials is still to be further studied. The development of novel composite biomaterials is a trend. ConclusionThe annulus fibrosus tissue engineering for the anulus fibrosus regeneration and repair will bring very broad prospects for clinical application in future.
ObjectiveTo investigate the feasibil ity and short-term effectiveness of percutaneous endoscopic spine surgery for treatment of lumbar disc herniation with posterior ring apophysis separation. MethodsBetween July 2008 and January 2013, 57 patients with lumbar disc herniation and posterior ring apophysis separation were treated. There were 39 males and 18 females, aged from 13 to 46 years (mean, 26.7 years). Of 57 cases, 29 had a clear trauma history. All patients had single segmental unilateral lumbar disc herniation, and the location was at L4, 5 in 22 cases and at L5, S1 in 35 cases; there were 25 cases of lateral bone fragments and 32 cases of central cortical and cancellous fragments. While fracture located at posterior inferior edge of the vertebrae at L4 level in 9 cases and at L5 level in 8 cases, at posterior superior edge at L5 level in 13 cases and in S1 level in 27 cases. Percutaneous endoscopic surgeries were performed via interlaminar or transforaminal approach from unilaterally symptomatic side for discectomy of lumbar disc herniation and partial or complete resection of free bone fragments. ResultsAccurate positioning was obtained in all patients during operation, and no complication of nerve root injury, hematoma formation, or dural tear occurred. The operation time was 20-85 minutes; the intraoperative fluoroscopy times were 2-15 times, and the blood loss was 3-10 mL. Postoperative radiographic examination showed that bone fragment was not removed in 16 cases, was partly removed in 32 cases, and was completely removed in 9 cases. Disc was completely removed. All patients were followed up 10-64 months (median, 16 months). According to modified Macnab criteria for the evaluation of effectiveness, the results were excellent in 48 cases, good in 6 cases, and fair in 3 cases, and the excellent and good rate was 94.7%. ConclusionPercutaneous endoscopic spine surgery through unilaterally symptomatic approach has advantages of small tissue damage and lumbar structure damage, less bleeding, and shorter recovery time. It has a good short-term effectiveness, but long-term effectiveness need further follow-up.
ObjectiveTo investigate the specific microRNA (miRNA) in osteogenic and chondrogenic differentiations of C3H10T1/2 cells. MethodsC3H10T1/2 cells were induced to differentiate into osteoblasts and chondrocytes.Specific miRNA more than 2 fold change and 2 average normalized probe signal between C3H10T1/2 and C3H10T1/2-derived osteoblast,and between C3H10T1/2 and C3H10T1/2-derived chondrocytes were screened out by miRNA microarray,and verified by real-time fluorescence quantitative PCR (RT-qPCR). ResultsAlkaline phosphatase expression of osteogenic induced group was significantly higher than that of control group at 7 days after induced (P<0.05).RT-qPCR results showed the expressions of Runx2,serine protease (Sp7),collagen type I,and osteopontin (OPN) genes were significantly increased at 7,14,and 21 days after induced when compared with before induced (P<0.05).Western blot results showed the expressions of Runx2,Sp7,collagen type I,and OPN proteins of osteogenic induced group were significantly higher than those of control group at 21 days after induced (P<0.05).The expressions of SOX9,collagen type Ⅱ,Aggrecan,and Has2 were significantly increased at 5,10,and 15 days after induced when compared with before induced (P<0.05).The expressions of SOX9,collagen type 2,Aggrecan,and Has2 proteins of chondrogenic induced group were significantly higher than those of control group at 15 days after induced (P<0.05).Totally,10 osteogenic and 3 chondrogenic miRNA more than 2 fold change and 2 average normalized probe signal were screened out by miRNA microarray.RT-qPCR results of these specific miRNAs were similar to microarray results except miR-455-3p. ConclusionSpecific miRNAs are screened out by microarray and it is a good foundation for the future study on miRNA functional verification and target gene prediction.
ObjectiveTo investigate whether miR-93-5p suppresses osteogenic differentiation of mouse mesenchymal stem cells (C3H10T1/2) by targeting Smad5, a predicted target in silicon. MethodsSmad5 3'-UTRluciferase vector (pmiR-RB-REPORTTM) was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-93-5p on Smad5 3'-UTR-luciferase activity to identify whether Smad5 was the target gene of miR-93-5p. miR-93-5p mimics (group M), miR-93-5p inhibitor (group In), miR-93-5p mimics negative control (group MC), and miR-93-5p inhibitor negative control (group InC) were transfected into the C3H10T1/2 cells, respectively, and followed by induction of osteogenic differentiation. After 48 hours, the real-time fluorescent quantitative PCR (qRTPCR) and Western blot assays were performed to detect the relative expressions of Smad5 mRNA and protein. At 14 days, to realize the regulation role of miR-93-5p in osteogenic differentiation, the extracellular calcium deposition during the osteogenesis of C3H10T1/2 cells was tested by Alizarin red staining. ResultsDual-luciferase reporter gene assay showed that miR-93-5p could combine with Smad5 mRNA 3'-UTR specificity, and inhibited its luciferase activity (P<0.05). After 48 hours, no significant difference was shown in the relative expression of Smad5 mRNA between group M and group MC as well as between group In and group InC by qRT-PCR assay (P>0.05); however, the results of Western blot assay showed that the relative expression of Smad5 protein was significantly decreased in group M and increased in group In when compared with groups MC and InC (P<0.05). At 14 days after osteogenic induction, Alizarin red staining showed that the extracellular calcium deposition of group M was obviously less than that of group MC, and it was obviously more in group In than in group InC. ConclusionSmad5 may be the target gene of miR-93-5p. And miR-93-5p can suppress osteogenic differentiation of C3H10T1/2 cells by directly targeting Smad5.
ObjectiveTo determine the feasibility and effectiveness of percutaneous endoscopic transforaminal discectomy (PETD) for recurrent lumbar disc herniation (RLDH). MethodsBetween June 2009 and December 2011, 56 patients with RLDH underwent PETD after local anesthesia, including 30 males and 26 females, with a mean age of 50 years (range, 24-70 years). The involved segments were L3, 4 in 3 cases, L4, 5 in 34 cases, and L5-S1 in 19 cases. Of 56 patients, 48 suffered from ipsilateral re-herniation, and 8 suffered from contralateral re-herniation. All the patients had a mean pain-free interval of 5.5 years (range, 6 months to 27 years). The visual analogue scale (VAS) score for back pain was 6.18±1.44 and the VAS score for leg pain was 7.66±1.03. Postoperative effectiveness was assessed based on the VAS score and modified MacNab criteria. ResultsThe mean operation time was 60.4 minutes (range, 30-100 minutes) and the mean duration of hospital stay was 5.1 days (range, 3-6 days). All patients were followed up 28.2 months on average (range, 24-56 months). Patients obtained immediate pain relief postoperatively. The postoperative VAS scores of back and leg pain at 1 month, 3 months, 12 months, and last follow-up were significantly decreased when compared with preoperative score (P<0.05). Based on the modified MacNab criteria, the results were excellent in 39 cases, good in 9 cases, fair in 5 cases, and poor in 3 cases at 12 months after operation, and the excellent and good rate was 85.7%. Surgery-related complications were found in 5 cases (8.9%); one patient (1.8%) suffered from recurrence at 18 months postoperatively, and the symptom was relieved after open lumbar discectomy and intervertebral fusion surgery. ConclusionPETD has several advantages in treating RLDH, such as avoiding from the old scar tissue, decreasing operation-related complications, shortening operation time, reducing trauma, and obtaining rapid postoperative recovery. This technique is feasible and effective for RLDH.
ObjectiveTo investigate the application and technical essentials of computer-assisted navigation in the surgical management of periacetabular fractures and pelvic fractures. MethodsBetween May 2010 and May 2011, 39 patients with periacetabular or anterior and posterior pelvic ring fractures were treated by minimally invasive fixation under computer-assisted navigation and were followed up more than 2 years, and the clinical data were analyzed retrospectively. There were 21 males and 18 females, aged 15-64 years (mean, 36 years). Fractures were caused by traffic accident in 23 cases, crush injury in 6 cases, and falling from height in 10 cases. Of them, 6 cases had acetabular fractures; 6 cases had femoral neck fractures; 18 cases had dislocation of sacroiliac joint; and 15 cases had anterior pelvic ring injuries. All patients were treated with closed or limited open reduction and screw fixations assisted with navigation. ResultsEighty-nine screws were inserted during operation, including 8 in the acetabulum, 18 in the neck of the femur, 33 in the sacroiliac joint, and 30 in the symphysis pubis and pubic rami. The mean time of screw implanted was 20 minutes (range, 11-38 minutes), and the average blood loss volume was 20 mL (range, 10-50 mL). The postoperative pelvic X-ray and three dimensional CT scan showed good reduction of fractures and good position of the screws. No incision infection, neurovascular injury, or implant failure occurred. All patients were followed up 27-33 months with an average of 29.6 months. The patients could walk with full weight loading at 6-12 weeks after operation (mean, 8 weeks); at last follow-up, the patients could walk on the flat ground, stand with one leg, and squat down, and they recovered well enough to do their job and to live a normal life. ConclusionMinimally invasive fixation under computer-assisted navigation may be an excellent method to treat some specific types of periacetabular and anterior and posterior pelvic ring fractures because it has the advantages of less trauma and blood loss, lower complication incidence, and faster recovery.
ObjectiveTo summarize the experience in management of occult cerebrospinal fluid leakage (CSFL) in posterior thoracolumbar surgeries, and to explore the best drainage duration, as well as to analyse the ways to reduce the risk of CSFL. MethodsA retrospective analysis was made on the clinical data of 26 patients with occult CSFL in posterior thoracolumbar surgeries between January 2011 and January 2013. There were 15 males and 11 females, with the average age of 48.7 years (range, 36-59 years). Headache occurred in 19 cases, and 5 cases had nausea with 3 cases also having vomiting after operation. Drainage tube unobstructed and no CSFL from the skin incision were observed in 23 cases at the postoperative 2nd day, and the drainage pipe clamp test was performed at the 3rd day. Twenty-one patients had no CSFL and were given extubation; 2 cases having CSFL were given extubation after conservative treatments for 10 days. Three patients had CSFL with ineffective conservative treatments at the postoperative 2nd day, then received reoperation, incision suture, and drainage. At the postoperative 3rd day, if no CSFL was observed, these patients were given extubation and stayed in bed for 3-5 days. ResultsAll incisions healed and the healing time was 7-15 days (mean, 8 days). No incision infection, persistent CSFL, and other complications occurred. After extubation, headache, nausea, vomiting, and other symptoms were alleviated immediately. All patients were followed up 12-24 months (mean, 16 months). MRI at the postoperative 6th month showed no subcutaneous epidural pseudocyst. ConclusionThe quality of suturing is the key factor to prevent occult CSFL in posterior thoracolumbar surgery. Under the premise of good suture quality, extubation can be given at the postoperative 3rd day. Before extubation, the drainage pipe clamp test can be performed to make sure no CSFL and to reduce the risk of CSFL from the surgical incision after extubation.
ObjectiveTo observe the character of spino-pelvic sagittal alignment in patients with high-grade L5 isthmic spondylolisthesis, and to analyze the sagittal alignment alteration after operation. MethodBetween January 2009 and June 2014, 25 patients with high-grade L5 isthmic spondylolisthesis underwent posterior surgery, and the clinical data were retrospectively analyzed as study group. There were 14 males and 11 females with a mean age of 42.5 years (range, 20-65 years). The mean disease duration was 6 months (range, 3-12 months). According to the Meyerding evaluating system, 15 cases were rated as degree III, and 10 cases as degree IV. Eighty healthy adult volunteers were recruited simultaneously as control group. The lumbar sacral angle (LSA), pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS), lumbar lordosis (LL), thoracic kyphosis (TK), and sagittal vertical axis (SVA) were measured on preoperative and last follow-up standing full length lateral X-ray films. And these parameters were compared between study group (preoperative parameters) and control group. Then the patients in study group were divided into 2 subgroups according to Hresko's method:the balanced pelvis subgroup (n=14) and unbalanced pelvis subgroup (n=11) . The sagittal parameters were compared between 2 subgroups, and the alteration of sagittal parameters before surgery and at last follow-up was analyzed in each subgroup respectively. ResultsAll patients in study group were followed up 18 months on average (range, 6-48 months). After surgery, spondylolisthesis was reduced from degree III to degree 0 in 12 cases and to degree I in 3 cases, and from degree IV to degree 0 in 6 cases and to degree I in 4 cases. Bone fusion was obtained in all patients at last follow-up. The preoperative PI, SS, PT, and SVA of study group were significantly greater than those of control group (P<0.05) , while the TK was significantly smaller than that of control group (P<0.05) . In the balanced pelvis subgroup, LSA, LL, and SVA at last follow-up significantly decreased while TK significantly increased when compared with preoperative ones (P<0.05) . In the unbalanced pelvis subgroup, LSA, PT, and SVA at last follow-up significantly decreased while SS, LL, and TK significantly increased when compared with preoperative ones (P<0.05) . The preoperative LSA and PT in the unbalanced pelvis subgroup were significantly greater, while SS, LL, and TK were significantly smaller than those of balanced pelvis subgroup (P<0.05) ; while at last follow-up, significant differentce was found only in LSA between 2 subgroups (P<0.05) . ConclusionsThe LSA should be paid more attention in surgery to assure recovery of the sagittal balance because patients with high-grade L5 isthmic spondylolisthesis have greater PI, abnormal lumbosacral kyphosis, and sagittal imbalance.