Lactoferrin Mediates Enhanced Osteogenesis of Adipose-Derived Stem Cells: Innovative Molecular and Cellular Therapy for Bone Repair.

A prospective source of stem cells for bone tissue engineering is adipose-derived stem cells (ADSCs), and BMP-2 has been proven to be highly effective in promoting the osteogenic differentiation of stem cells. Rarely has research been conducted on the impact of lactoferrin (LF) on ADSCs' osteogenic differentiation. As such, in this study, we examined the effects of LF and BMP-2 to assess the ability of LF to stimulate ADSCs' osteogenic differentiation. The osteogenic medium was supplemented with the LF at the following concentrations to culture ADSCs: 0, 10, 20, 50, 100, and 500 µg/mL. The Cell Counting Kit-8 (CCK-8) assay was used to measure the proliferation of ADSCs. Calcium deposition, alkaline phosphatase (ALP) staining, real-time polymerase chain reaction (RT-PCR), and an ALP activity assay were used to establish osteogenic differentiation. RNA sequencing analysis was carried out to investigate the mechanism of LF boosting the osteogenic development of ADSCs. In the concentration range of 0-100 µg/mL, LF concentration-dependently increased the proliferative vitality and osteogenic differentiation of ADSCs. At a dose of 500 µg/mL, LF sped up and enhanced differentiation, but inhibited ADSCs from proliferating. LF (100 and 500 µg/mL) produced more substantial osteoinductive effects than BMP-2. The PI3 kinase/AKT (PI3K/AKT) and IGF-R1 signaling pathways were significantly activated in LF-treated ADSCs. The in vitro study results showed that LF could effectively promote osteogenic differentiation of ADSCs by activating the PI3K/AKT and IGF-R1 pathways. In our in vitro investigation, an LF concentration of 100 µg/mL was optimal for osteoinduction and proliferation. Our study suggests that LF is an attractive alternative to BMP-2 in bone tissue engineering. As a bioactive molecule capable of inducing adipose stem cells to form osteoblasts, LF is expected to be clinically used in combination with biomaterials as an innovative molecular and cellular therapy to promote bone repair.

[Surgical treatment of delayed spinal cord injury caused by atypical compression of old thoracolumbar fractures].

OBJECTIVE: To explore the clinical characteristics and surgical treatment strategies of delayed spinal cord injury (SCI) caused by atypical compression of old thoracolumbar fracture. METHODS: Between January 2011 and June 2018, 32 patients with delayed SCI caused by atypical compression of old thoracolumbar fracture who met the inclusion criteria were admitted and divided into group A (20 cases, underwent anterior subtotal vertebral body resection+titanium mesh reconstruction+screw rod internal fixation) and group B (12 cases, underwent posterior 270° ring decompression of vertebral canal+titanium mesh reconstruction+screw rod internal fixation) according to the different operation approaches. There was no significant difference between the two groups in age, gender, cause of injury, fracture segment, disease duration, preoperative American Spinal Injury Association (ASIA) classification, and preoperative back pain visual analogue scale (VAS) score, lumbar Japanese Orthopaedic Association (JOA) score, kyphosis angle, and vertebral canal occupational ratio ( P>0.05). The incision length, operation time, intraoperative blood loss, complications, and bone fusion time of reconstructed vertebrae were recorded and compared between the two groups; the kyphosis angle, back pain VAS score, and lumbar JOA score were used to evaluate the effectiveness. RESULTS: Except that the incision length in group A was significantly shorter than that in group B ( t=-4.865, P=0.000), there was no significant difference in intraoperative blood loss and operation time between the two groups ( P>0.05). There was no deaths or postoperative paraplegia cases in the two groups, and no deep infection or skin infection occurred. There was 1 case of cerebrospinal fluid leakage, 1 case of inferior vena cava injury, and 1 case of chyle leakage in group A. No serious complications occurred in group B. There was no significant difference in the incidence of complications between the two groups ( P=0.274). All 32 patients were followed up 12-61 months, with an average of 20.8 months. The follow-up time for groups A and B were (19.35±5.30) months and (23.25±12.20) months respectively, and the difference was not significant ( t=-1.255, P=0.219). The reconstructed vertebrae in all cases obtained bony fusion postoperatively. The fusion time of groups A and B were (8.85±2.27) months and (8.50±2.50) months respectively, and the difference was not significant ( t=0.406, P=0.688). The kyphosis angle, back pain VAS score, and lumbar JOA score of the two groups at each time point after operation and last follow-up were significantly improved when compared with preoperatively ( P<0.05); the lumbar JOA score was further improved with time postoperatively ( P<0.05), while the kyphosis angle and the VAS score of back pain remained similarly ( P>0.05). Comparison of kyphosis angle, back pain VAS score, and lumbar JOA score between the two groups at various time points postoperatively showed no significant difference ( P>0.05). At last follow-up, the JOA score improvement rate in groups A and B were 83.87%±0.20% and 84.50%±0.14%, respectively, and the difference was not significant ( t=-0.109, P=0.914); the surgical treatment effects of the two groups were judged to be significant. CONCLUSION: In the later stage of treatment of old thoracolumbar fractures, even mild kyphosis and spinal canal occupying may induce delayed SCI. Surgical correction and decompression can significantly promote the recovery of damaged spinal cord function. Compared with anterior approach surgery, posterior approach surgery has the advantages of less trauma, convenient operation, and fewer complications, so it can be preferred.

Effects of Andersson lesion treatment in ankylosing spondylitis: A medical record review study focused on medium- to long-term outcomes.

The present study aimed to evaluate the clinical efficacy of Andersson lesion (AL) treatments and prognostic factors using medium- to long-term follow-up data and discuss the clinical characteristics and treatment of AL. Forty-eight consecutive AL cases at our center from June 2011 to March 2018 were retrospectively analyzed, including 13 cases treated conservatively and 35 treated surgically. Epidemiological characteristics, treatment modalities, clinical features and outcomes, and prognostic factors of the Japanese Orthopaedic Association (JOA) recovery rate were reviewed. Neurological functional recovery was evaluated by American Spinal Injury Association (ASIA) classification. Clinical efficacy was evaluated by JOA score, visual analog scale (VAS) score, and Cobb's angle. The mean overall follow-up duration was 44.5±18.5 months (range, 27-85 months). There were 36 male and 12 female patients, with a mean age of 49.4±13.1 years (range, 26-72 years). The most common lesion location was the thoracolumbar region, i.e., T10-L2 (n=33; 68.8%), followed by the thoracic (n=10) and lumbar (n=5) regions. Patients treated surgically showed significantly better JOA scores, VAS scores and Cobb's angles at the final follow-up than did patients treated conservatively (P<.05). Univariate and binary logistic regression analyses identified two prognostic factors of the JOA score recovery rate: treatment modality (OR=0.157; 95%CI, 0.028-0.89; P=.036) and bone fusion (OR=9.965; 95%CI, 2.052-48.387; P=.004). Conservative treatment and bone nonunion predict worse JOA score recovery. Surgery remains the optimal treatment for AL in ankylosing spondylitis patients, with better clinical efficacy demonstrated by medium- to long-term follow-up data.

Lumbopelvic Fixation and Sacral Decompression for U-shaped Sacral Fractures: Surgical Management and Early Outcome.

U-shaped sacral fractures are rare and often difficult to diagnose primarily due to the difficulty in obtaining adequate imaging and the severe associated injuries. These fractures are highly unstable and frequently cause neurological deficits. The majority of surgeons have limited experience in management of U-shaped sacral fractures. No standard treatment protocol for U-shaped sacral fractures has been available till now. This study aimed to examine the management of U-shaped sacral fractures and the early outcomes. Clinical data of 15 consecutive patients with U-shaped sacral fracture who were admitted to our trauma center between 2009 and 2014 were retrospectively analyzed. Demographics, fracture classification, mechanism of injury and operative treatment and deformity angle were assessed. All the patients were treated with lumbopelvic fixation or (and) sacral decompression. EQ-5d score was applied to evaluate the patients' quality of life. Of the 15 consecutive patients with U-shaped sacral fracture, the mean age was 28.8 years (range: 15-55 years) at the time of injury. There were 6 females and 9 males. The mean followup time was 22.7 months (range: 9 47 months) and mean full weight-bearing time was 9.9 weeks (range: 8-14 weeks). Ten patients received lumbopelvic fixation and sacral decompression, one lombosacral fixation, and 4 merely sacral decompression due to delayed diagnosis or surgery. The post-operation deformity angle (mean 27.87°, and range: 8°-90°) of the sacrum was smaller than that pre-operation (mean 35.67; range: 15-90) with no significance difference noted. At the latest follow-up, all patients obtained neurological recovery with different extents. Visual analogue score (VAS) was reduced from preoperative 7.07 (range: 5-9) to postoperetive 1.93 (range: 1-3). All patients could walk without any aid after treatment. Eight patients were able to care for themselves and undertook some daily activities. Five patients had returned to work foil time. In conclusion, lumbopelvic fixation is an effective method for stabilization of U-shaped sacral fractures with fewer complications developed. Effective reduction and firm fixation are the prerequisite of early mobilization and neurological recovery. Sacral decompression effectively promotes neurological recovery even in patients with old U-shaped sacral fractures.

MicroRNA-449a upregulation promotes chondrocyte extracellular matrix degradation in osteoarthritis.

Osteoarthritis (OA) is a highly prevalent chronic and degenerative joint disease characterized by the continuous destruction of the articular cartilage. MicroRNAs (miRNAs) have been reported to be strongly involved in the pathogenesis of OA. The aim of this study was to explore whether miR-449a regulates the expression of growth differentiation factor5 (GDF5), which promotes chondrocyte extracellular matrix (ECM) degradation. We found that miR-449a expression was upregulated in OA cartilage compared to that in normal cartilage. Overexpression of miR-449a increased the expression of chondrocyte ECM catabolic factors, such as matrix metalloproteinases and a disintegrin and metalloproteinase with thrombospondin motif, while inhibiting that of anabolic genes, such as type II collagen and aggrecan. In contrast, suppression of miR-449a exerted the opposite effects. Moreover, GDF5 was identified as a direct target of miR-449a, and its expression was significantly suppressed by miR-449a overexpression. In addition, the suppression of chondrocyte ECM degradation induced by miR-449a inhibitor was attenuated by small interfering RNA-mediated knockdown of GDF5. Overall, these results suggest that miR-449a contributes to chondrocyte ECM degradation in OA via directly targeting GDF5, thereby providing insights to a promising therapeutic target for the treatment of human OA.

The Diagnostic and Prognostic Role of Interleukin 12B and Interleukin 6R Gene Polymorphism in Patients With Ankylosing Spondylitis.

OBJECTIVES: Interleukin 23 (IL-23) pathway and IL-1 cluster genes play prominent role in the etiopathology of ankylosing spondylitis (AS). The aim of this study was to investigate the diagnostic and prognostic role of 5 single-nucleotide polymorphisms related to IL-23 pathway and IL-1 cluster genes in AS patients. METHODS: Four hundred thirty-one patients with AS and 206 age- and sex-matched healthy controls were recruited in this prospective cohort study. Five potential single-nucleotide polymorphisms (IL-23R [rs11209026], IL-12B [rs6871626], TYK2 [rs6511701], IL-6R [rs4129267], and IL-1R2 [rs2192752]) related to IL-23 pathway and IL-1 cluster genes by analyzing previous studies were genotyped. Among 431 total AS patients, 198 active cases were treated and followed up for 24 weeks. RESULTS: Frequencies of IL-12B AA (rs6871626) and IL-6R TT (rs4129267) genotypes were increased in AS patients compared with healthy controls (both P < 0.001), and IL-12B A (rs6871626) as well as IL-6R T (rs4129267) allele increased the risk of AS independently (both P < 0.001). The Bath Ankylosing Spondylitis Disease Activity Index score was found to be elevated in AS patients with IL-12B AA (rs6871626) compared with patients with the CA and CC genotypes (P = 0.002 and P < 0.001, respectively), and the Bath Ankylosing Spondylitis Functional Index score was also increased in AS patents with IL-12B AA (rs6871626) than in those with the CA and CC genotypes (P = 0.001 and P < 0.001). In addition, IL-6R T (rs4129267) allele could predict a worse ASAS-20 (Assessment of SpondyloArthritis international Society) response at week 24 as an independent factor by multivariate logistic regression analysis with additive model (P = 0.011). CONCLUSIONS: Interleukin 12B (rs6871626) and IL-6R (rs4129267) gene polymorphisms could serve as promising biomarkers for diagnosis and prognosis in AS patients.

Comparison of percutaneous endoscopic lumbar discectomy versus open lumbar microdiscectomy for lumbar disc herniation: A meta-analysis.

PURPOSE: The purpose of the study is to perform a systematic review and meta-analysis to evaluate the clinical results of percutaneous endoscopic lumbar discectomy (PELD) and open lumbar microdiscectomy (OLM) for the treatment of lumbar disc herniation (LDH). METHODS: Randomized controlled trials or non-randomized controlled trials published from the time when databases were built to March 2016 that compared the clinical effectiveness of PELD and OLM surgical approaches for the treatment of LDH were acquired by a comprehensive search in four electronic databases (PubMed, EMBASE, Web of Science and Cochrane library). A total of 7 studies (1389 patients) were included in this systematic review and meta-analysis. Pooled mean differences (MD) and odds ratios (OR) and with 95% CIs were calculated for the outcomes. RESULT: The results showed that there were no statistically between the PELD group and OLM group in terms of preoperative VAS-BP score (WMD = 0.03; 95% CI: -0.99 to 1.05; P = 0.95), postoperative VAS-BP score (WMD = -0.56; 95% CI: -1.43 to 0.31; P = 0.21), postoperative ODI (WMD = -0.98; 95% CI: -4.96 to 3.00; P = 0.63), complication rate (OR = 1.79; 95% CI: 0.95 to 3.37; P = 0.07) or reoperation rate (OR = 1.44; 95% CI: 0.94 to 2.20; P = 0.09). PELD group was associated with shorter operation time (WMD = -12.83; 95% CI: -24.79 to -0.87; P = 0.04) and hospital stay (WMD = -5.49; 95% CI: -8.63 to -2.35; P = 0.0006). CONCLUSION: The existing evidence indicate that no superiority exists between the two surgical approaches for the treatment of LDH in terms of functional outcome, complication rate and reoperation rate, in spite of that PELD surgical group can achieve shorter operation time and hospital stay than OLM surgical group.

Effect of vacuum-assisted closure combined with open bone grafting to promote rabbit bone graft vascularization.

BACKGROUND: Patients with composite bone non-union and soft tissue defects are difficult to treat. Vacuum-assisted closure (VAC) combined with open bone grafting is one of the most effective treatments at present. The aim of the present study was to preliminarily investigate the effect and mechanism of VAC combined with open bone grafting to promote rabbit bone graft vascularization, and to propose a theoretical basis for clinical work. MATERIAL/METHODS: Twenty-four New Zealand white rabbits were randomly divided into an experimental and a control group. Allogeneic bones were grafted and banded with the proximal femur with a suture. The experimental group had VAC whereas the control group had normal wound closure. The bone vascularization rate was compared based on X-ray imaging, fluorescent bone labeling (labeled tetracycline hydrochloride and calcein), calcium content in the callus, and expression of fibroblast growth factor-2 (FGF-2) in bone allografts by Western blot analysis at the 4th, 8th, and 12th week after surgery. RESULTS: At the 4th, 8th, and 12th week after surgery, the results of the tests demonstrated that the callus was larger, contained more calcium (p<0.05), and expressed FGF-2 at higher levels (p<0.05) in the experimental group than in the control group. Fluorescent bone labeling showed the distance between the two fluorescent ribbons was significantly shorter in the control group than in the experimental group at the 8th and 12th week after surgery. CONCLUSIONS: VAC combined with open bone grafting promoted rabbit bone graft vascularization.

One-stage reconstruction with open bone grafting and vacuum-assisted closure for infected tibial non-union.

INTRODUCTION: Non-union of the tibia complicated by osteomyelitis is one of the most challenging problems in orthopaedic surgery. There remains a significant amount of debate and controversy regarding the optimal medical management of infected tibial non-union. There are few articles which have reported the outcomes of treatment for infected non-union of tibia from single-stage reconstruction with open bone grafting plus vacuum-assisted closure (VAC). MATERIAL AND METHODS: Our report covers experience between March 2007 and February 2010 of open bone grafting plus VAC in one stage for patients with infected tibial non-union. The time for bone union and wound healing to occur, the duration of hospitalisation, and the rate of resolution of infection were all analysed. The main outcome measures were based on a clinical scoring system that assessed functional ability, range of knee and ankle motion, shortening, infection and pain. Fifteen patients were involved in this study. RESULTS: All patients were followed up for an average of 22.6 months (range: 14-42 months). Bone union was achieved in 93.3% (14/15) of patients after a mean of 5.93 months (range: 3-10 months). All wounds healed within an average period of 5 weeks (range: 3-10 weeks), and the function and appearance of all limbs were satisfactory. CONCLUSIONS: Open bone grafting combined with VAC in a one-stage procedure can be a feasible alternative to the treatment of infected tibial non-union, especially for those wounds which are not good candidates for microsurgery; however, further studies are required to confirm the likely benefits.

The retrospective analysis of posterior short-segment pedicle instrumentation without fusion for thoracolumbar burst fracture with neurological deficit.

This study aims to investigate the efficacy of posterior short-segment pedicle instrumentation without fusion in curing thoracolumbar burst fracture. All of the 53 patients were treated with short-segment pedicle instrumentation and laminectomy without fusion, and the restoration of retropulsed bone fragments was conducted by a novel custom-designed repositor (RRBF). The mean operation time and blood loss during surgery were analyzed; the radiological index and neurological status were compared before and after the operation. The mean operation time was 93 min (range: 62-110 min) and the mean intraoperative blood loss was 452 mL in all cases. The average canal encroachment was 50.04% and 10.92% prior to the surgery and at last followup, respectively (P < 0.01). The preoperative kyphotic angle was 17.2 degree (± 6.87 degrees), whereas it decreased to 8.42 degree (± 4.99 degrees) at last followup (P < 0.01). Besides, the mean vertebral body height increased from 40.15% (± 9.40%) before surgery to 72.34% (± 12.32%) at last followup (P < 0.01). 45 patients showed 1-2 grades improvement in Frankel's scale at last followup. This technique allows for satisfactory canal clearance and restoration of vertebral body height and kyphotic angle, and it may promote the recovery of neurological function. However, further research is still necessary to confirm the efficacy of this treatment.

TNF/TNFR signal transduction pathway-mediated anti-apoptosis and anti-inflammatory effects of sodium ferulate on IL-1ß-induced rat osteoarthritis chondrocytes in vitro.

INTRODUCTION: Sodium ferulate (SF) is a natural component of traditional Chinese herbs. Our previous study shows that SF has a protective effect on osteoarthritis (OA). The objective of this study was to investigate the effect of SF on the TNF/TNF receptor (TNFR) signal transduction pathway of rat OA chondrocytes. METHODS: Primary rat articular chondrocytes were co-treated with IL-1ß and SF. Chondrocyte apoptosis was assessed by fluorescein isothiocyanate-annexin V/propidium iodide assay. The PCR array was used to screen the expression of 84 key genes involved in apoptosis. The release of TNFα and prostaglandin E2 were analyzed by ELISA. Expressions of proteins were assessed by western blotting. The activity of NF-κB was determined by electrophoretic mobility shift assay (EMSA). Gene expression of inducible nitric oxide synthase (iNOS) was evaluated by real-time quantitative PCR. The nitric oxide content was measured with the Griess method. RESULTS: After treatment with SF, the apoptosis rate of chondrocytes significantly attenuated (P < 0.01). Results of the apoptosis PCR array suggested that mRNA expression of some core proteins in the TNF/TNFR pathway showed valuable regulation. The protein expressions of TNFα, TNFR-1, TNF receptor-associated death domain, caspase-8 and caspase-3 were prevented by SF in a concentration-dependent manner. SF also inhibited activities of caspase-8 and caspase-3 compared with the OA model control (P < 0.01). TNF receptor-associated factor-2 expression, phosphorylations of inhibitor of NF-κB kinase (IKK) subunits alpha and beta, and NF-κB inhibitor, alpha (IκBα) were all concentration-dependently suppressed by SF treatment. The results of EMSA showed that SF inhibited the activity of NF-κB. In addition, the expressions of cycloxygenase-2 and iNOS and the contents of prostaglandin E2 and NO were attenuated with the treatment of SF (P < 0.01). CONCLUSION: SF has anti-apoptosis and anti-inflammatory effects on an OA model induced by IL-1ß in vitro, which were due to inhibitory actions on the caspase-dependent apoptosis pathway and the IKK/NF-κB signal transduction pathway of the TNF/TNFR pathway.

Concurrent access to sucrose pellets decreases methamphetamine-seeking behavior in Lewis rats.

Investigation of the role of choice between use of drugs of abuse and pursuit of alternative non-drug reinforcers is receiving greater attention. An understanding of the determinants influencing choice between drugs and alternative reinforcers will eventually lead to an understanding to the neural substrates of the drug altered brain. We investigated the impact of concurrent access to sucrose pellets on methamphetamine self-administration and self-regulated reinstatement of methamphetamine seeking following extinction training in Lewis rats. Our results from the self-administration experiment show that rats with concurrent access to sucrose self-administered significantly less methamphetamine compared to the methamphetamine only group. For our extinction/reinstatement experiment, concurrent access to sucrose during self-regulated methamphetamine reinstatement reduced methamphetamine intake and non-reinforced methamphetamine-seeking behavior in rats compared to rats that received access to just methamphetamine. These findings indicate that concurrent access to alternative reinforcers during various stages of methamphetamine-seeking behavior robustly decreased methamphetamine intake and serves as a valid rodent choice paradigm.

Contributions of nucleus accumbens core and shell GluR1 containing AMPA receptors in AMPA- and cocaine-primed reinstatement of cocaine-seeking behavior.

Glutamate signaling in the nucleus accumbens influences reinstatement of previously extinguished cocaine-seeking behavior in rats. Whether or not region specific glutamate signaling in the nucleus accumbens contributes to reinstatement of cocaine-seeking behavior is not known. We investigated whether directly stimulating ionotropic glutamate receptors (GluRs) within the nucleus accumbens core or shell would differentially influence renewed cocaine-seeking behavior following extinction training. We also tested the hypothesis that GluR1 subunit (GluR1) containing alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid (AMPA) receptors in the nucleus accumbens core and not the shell regulate reinstatement of previously extinguished cocaine-seeking behavior. Microinjection of AMPA into the nucleus accumbens shell and the nucleus accumbens core dose-dependently elicited significant cocaine-seeking behavior. Administration of antisense oligonucleotides (AS) directed against GluR1 subunit mRNA into the core and shell disrupted AMPA- and cocaine-primed reinstatement--with the most pronounced effects seen in the nucleus accumbens shell. These results demonstrate that GluRs in the nucleus accumbens core and shell influence AMPA- and cocaine-primed reinstatement, yet the nucleus accumbens shell exerts a prepotency over the nucleus accumbens core.