Investigation of Metabolic and Inflammatory Disorder in the Aging FGF21 Knockout Mouse.

Aging is a physiological condition accomplished with persistent low-grade inflammation and metabolic disorders. FGF21 has been reported to act as a potent longevity determinant, involving inflammatory response and energy metabolism. In this study, we engineered aging FGF21 knockout mice of 36-40 weeks and observed that FGF21 deficiency manifests a spontaneous inflammatory response of lung and abnormal accumulation of lipids in liver. On one hand, inflamed state in lungs and increased circulating inflammatory cytokines were found in FGF21 knockout mice of 36-40 weeks. To evaluate the ability of FGF21 to suppress inflammation, a subsequent study found that FGF21 knockout aggravated LPS-induced pulmonary exudation and inflammatory infiltration in mice, while exogenous administration of FGF21 reversed these malignant phenotypes by enhancing microvascular endothelial junction. On the other hand, FGF21 knockout induces fatty liver in aging mice, characterized by excessive accumulation of triglycerides within hepatocytes. Further quantitative metabolomics and lipidomics analysis revealed perturbed metabolic profile in liver lacking FGF21, including disrupted glucose and lipids metabolism, glycerophospholipid metabolism, and amino acid metabolism. Taken together, this investigation reveals the protective role of FGF21 during aging by weakening the inflammatory response and balancing energy metabolism.

Biomimetic Structural Protein Based Magnetic Responsive Scaffold for Enhancing Bone Regeneration by Physical Stimulation on Intracellular Calcium Homeostasis.

The bone matrix has distinct architecture and biochemistry which present a barrier to synthesizing bone-mimetic regenerative scaffolds. To mimic the natural structures and components of bone, biomimetic structural decellularized extracellular matrix (ECM)/regenerated silk fibroin (RSF) scaffolds incorporated with magnetic nanoparticles (MNP) are prepared using a facile synthetic methodology. The ECM/RSF/MNP scaffold is a hierarchically organized and interconnected porous structure with silk fibroin twined on the collagen nanofibers. The scaffold demonstrates saturation magnetization due to the presence of MNP, along with good cytocompatibility. Moreover, the ß-sheet crystalline domain of RSF and the chelated MNP could mimic the deposition of hydroxyapatite and enhance compressive modulus of the scaffold by ≈20%. The results indicate that an external static magnetic field (SMF) with a magnetic responsive scaffold effectively promotes cell migration, osteogenic differentiation, neogenesis of endotheliocytes in vitro, and new bone formation in a critical-size femur defect rat model. RNA sequencing reveals that the molecular mechanisms underlying this osteogenic effect involve calsequestrin-2-mediated Ca2+ release from the endoplasmic reticulum to activate Ca2+ /calmodulin/calmodulin-dependent kinase II signaling axis. Collectively, bionic magnetic scaffolds with SMF stimulation provide a potent strategy for bone regeneration through internal structural cues, biochemical composition, and external physical stimulation on intracellular Ca2+ homeostasis.

Small non-coding RNAome changes during human chondrocyte senescence as potential epigenetic targets in age-related osteoarthritis.

Chondrocyte senescence is a decisive component of age-related osteoarthritis, however, the function of small noncoding RNAs (sncRNAs) in chondrocyte senescence remains underexplored. Human hip joint cartilage chondrocytes were cultivated up to passage 4 to induce senescence. RNA samples were extracted and then analyzed using small RNA sequencing and qPCR. ß-galactosidase staining was used to detect the effect of sncRNA on chondrocyte aging. Results of small RNA sequencing showed that 279 miRNAs, 136 snoRNAs, 30 snRNAs, 102 piRNAs, and 5 rasiRNAs were differentially expressed in senescent chondrocytes. The differential expression of 150 sncRNAs was further validated by qPCR. Transfection of sncRNAs and ß-galactosidase staining were also performed to further revealed that hsa-miR-135b-5p, SNORA80B-201, and RNU5E-1-201 have the function to restrain chondrocyte senescence, while has-piR-019102 has the function to promote chondrocyte senescence. Our data suggest that sncRNAs have therapeutic potential as novel epigenetic targets in age-related osteoarthritis.

Bioabsorbable self-retaining PLA/nano-sized ß-TCP cervical spine interbody fusion cage in goat models: an in vivo study.

STUDY DESIGN: This is an experimental animal study. OBJECTIVE: The objective of this study was to compare an anterior cervical discectomy and interbody fusion of a novel polylactide/nano-sized ß-tricalcium phosphate (PLA/nß-TCP) bioabsorbable self-retaining cervical fusion cage (BCFC) with an autologous bone graft and polyetheretherketone (PEEK) cages. BACKGROUND: Although PLA cervical cages have potential advantages compared with traditional materials, they are not currently routinely used in spine surgery because of undesirable effects such as the lack of osteoconductivity and osteolysis around the implant. This study involved the manufacturing of a bioabsorbable cage from PLA/nß-TCP that was then used as a device for anterior cervical discectomy and fusion (ACDF) on a goat cervical spine fusion model. MATERIALS AND METHODS: Eighteen goats underwent C3/C4 discectomy and were randomly divided into three groups based on the following methods: Group A (n=6), an autologous bone graft; Group B (n=6), PEEK cage filled with an autologous graft; and Group C (n=6), BCFC filled with an autologous iliac bone. Radiography was performed preoperatively and postoperatively and at 1, 4, 8, and 12 weeks after the operation. Disc space height (DSH) was measured at the same time. After 12 weeks, the fused segments were harvested and evaluated with functional radiographic views, biomechanical testing, and histological analyses. RESULTS: Over a 12-week period, the BCFC and PEEK cage groups exhibited significantly higher DSH values than the bone graft group. Additionally, the BCFC group yielded a significantly lower range of motion in axial rotation than both the autologous bone graft and PEEK cage groups. A histologic evaluation revealed an increased intervertebral bone volume/total volume ratio and better interbody fusion in the BCFC group than in the other groups. CONCLUSION: The BCFC device exhibited better results than the autologous bone graft and PEEK cages in single-level ACDF models in vivo. This device may be a potential alternative to the current PEEK cages.

Single-Level Rigid Fixation Combined with Coflex: A Biomechanical Study.

BACKGROUND: The purpose of this biomechanical in vitro study was to compare the kinematics and intradiscal pressure achieved with 2 methods: L4-L5 pedicle screw-rod fixation (PSRF) with an upper L3-L4 Coflex device and L4-L5 PSRF alone. The results were used to characterize the biomechanics of the topping-off operation with a Coflex device for the lumbar motion segment adjacent to single-level rigid fixation. MATERIAL/METHODS: Six human cadaveric spine specimens were biomechanically tested in vitro (6 males, 0 females). The 3-dimensional specimen motion in response to applied loads during flexibility tests was determined. Loads were applied along anatomic axes to induce flexion-extension, lateral bending, and axial rotation. All specimens were first studied with intact lumbar motion segments, then with L4-L5 PSRF alone, and finally with L4-L5 PSRF with an upper L3-L4 Coflex device. A non-paired comparison of the 3 configurations under 3 different conditions was made. RESULTS: PSRF, with or without a Coflex device, significantly increased the range of motion (ROM) in the upper adjacent motion segments in all directions of loading. The intradiscal pressure (IDP) changed slightly. A correlation analysis showed that the ROM and IDP are significantly positively correlated. The application of the upper motion segment of the Coflex device provided greater stability in all directions of motion than did PSRF alone, particularly for extension (p<0.05), while use of a Coflex device did not significantly decrease the IDP compared with PSRF alone (p>0.05). CONCLUSIONS: These results suggest that L4-L5 PSRF with an L3-L4 Coflex device is more stable than L4-L5 PSRF alone. PSRF with an upper Coflex device is a promising alternative to PSRF alone. Based on these biomechanical tests, it might be considered a protective method to prevent adjacent segment degeneration (ASD), although some limitations with this in vitro study must be addressed in the future.

Decreased Zn(2+) Influx Underlies the Protective Role of Hypoxia in Rat Nucleus Pulposus Cells.

Zn(2+) is an essential component of metalloproteinases, and is required for their activity in cartilage; however, the effect of Zn(2+) on nucleus pulposus (NP) cells has not been widely investigated. The aim of this paper was to investigate the effect of intracellular Zn(2+) concentration ([Zn(2+)]i) in hypoxia-induced regulation of metalloproteinases (MMPs) and extracellular matrix (ECM) production in NP cells. NP cells from Sprague-Dawley (SD) rats were cultured as monolayers or in alginate beads. [Zn(2+)]i was assayed by FluoZin-3 AM staining. Alcian Blue staining, immunochemistry, 1,9-dimethylmethylene blue (DMMB) assay, and real-time PCR were used to assay collagen II, proteoglycan, and COL2A1, MMP-13, and ADAMTS-5 mRNA expression. ZIP8, a main Zn(2+) transporter in chondrocytes, was assayed by immunochemistry and in Western blotting. Interleukin (IL)-1ß- and ZnCl2-induced increases of [Zn(2+)]i were significantly inhibited by hypoxia. Hypoxia did not reverse a decline of ECM expression caused by IL-1ß and ZnCl2 in monolayer cultures, but did significantly attenuate the decreases of proteoglycan, glycosaminoglycan (GAG), and COL2A1 mRNA expression following IL-1ß and ZnCl2 treatment in alginate bead cultures. However, ZnCl2 inhibited the protective effect of hypoxia. Both an intracellular Zn(2+) chelator and hypoxia prevented the increase in MMP-13 mRNA expression. IL-1ß and ZnCl2 treatment increased ZIP8 expression in NP cells, and hypoxia inhibited ZIP8 expression. In conclusion, decrease of Zn(2+) influx mediates the protective role of hypoxia on ECM and MMP-13 expression. Consequently, changes in intracellular Zn(2+) concentration maybe involved in intervertebral disc degeneration.

[Comparison of percutaneous versus open monosegment instrumentation in the treatment of incomplete thoracolumbar burst fracture].

OBJECTIVE: To compare the safety and efficacies of traditional open versus percutaneous monosegmental pedicle screw fixation in the treatment of incomplete thoracolumbar spinal fracture. METHODS: A retrospective analysis was conducted for 44 inpatients with a diagnosis of incomplete thoracolumbar spinal fracture (AO classification: A3.1 and A3.2) undergoing monosegmental pedicle instrumentation (MSPI) from September 2008 to January 2011. There were 24 cases in percutaneous group and 20 cases in traditional open group. The mean operative duration, blood loss, blood drainage, visual analogue scale/score (VAS) and vertebral kyphotic angle at pre- and post-operation were evaluated. RESULTS: No significant differences existed in operative durations between two groups (P > 0.05). Significant differences between two groups were observed in terms of intra-operative blood loss and VAS scores at Week 1 postoperation (P < 0.05). There were no significant differences in VAS score preoperation, 1 year postoperation or pre-and post-operative vertebral kyphotic angle (P > 0.05). No complications of iatrogenic neurological injury or hardware failure occurred. CONCLUSION: The application of percutaneous monosegment pedicle instrumentation in the treatment of thoracolumbar fractures in type of A 3.1 and A 3.2 is both feasible and safe. Its postoperative therapeutic effect is comparable to that of traditional open monosegmental fixation.

[The treatment of monosegmental fixation and short-segment fixation on thoracolumbar burst fracture a retrospective controlled study].

OBJECTIVE: To investigate the safety and therapeutic effects of monosegment pedicle instrumentation in treating incomplete thoracolumbar burst fracture. METHODS: A retrospective analysis was conducted on 56 inpatients with incomplete thoracolumbar burst fracture (AO classification: A3.1 and A3.2) from April 2005 to January 2010. There were 28 cases were fixed with monosegment pedicle instrumentation (MSPI), 28 cases were fixed with short segment pedicle instrumentation (SSPI). The operative time, blood loss, visual analogue scale (VAS) and vertebral kyphotic angle (VK) before and after surgery were evaluated. RESULTS: In the group of MSPI, the mean operative time was (93 ± 20) min; the intraoperative blood loss was (184 ± 64) ml; the VK angle was 17° ± 10° before operation, 7° ± 7° at one week after operation, and 10° ± 7° at latest follow-up; VAS score was 7.6 ± 1.5 before operation, 2.4 ± 0.8 at one week after operation, and 1.5 ± 0.9 at latest follow-up; no adjacent segment degeneration was found. In the group of SSPI, the operative time was (102 ± 30) min; the intraoperative blood loss was (203 ± 88) ml; the VK angle was 17° ± 9° before operation, 7° ± 7° at one week after operation, and 8° ± 5° at latest follow-up; VAS score was 6.8 ± 1.3 before operation, 3.1 ± 0.5 at one week after operation, and 1.2 ± 0.7 at latest follow-up. One case of adjacent segment degeneration was found in 36 months after operation. There were no significantly statistical differences between two groups in operative time, blood loss, VAS score and VK angle before and after surgery (P > 0.05). The VAS score and VK angle at one week after surgery and latest follow-up all decreased obviously than preoperative ones in both groups (P < 0.05). CONCLUSIONS: MSPI for incomplete thoracolumbar burst fracture is effective and safe. The operative blood loss, the mean operative time, the improvement of VAS score and the VK angle in group MSPI are equal to those in group SSPI.

[The application of transgenic plant in evaluating the genotoxicity of environmental contaminants].

Environmental contaminants are powerful mutagenic factors for organisms. Several testing materials and methods have been used to assess the genotoxicity of environmental factors. Transgenic plants testing system can not only evaluate the level of genotoxicity, but also provide information on the genetic effects at molecular level. We introduce the use of transgenic plants in biomonitoring environmental factors.

Animal study on expression of laminin and fibronectin in cornea during wound healing following alkali burn.

OBJECTIVE: To observe the expression of laminin and fibronectin in alkali-burned corneas in rats. METHODS: A total of 18 normal Wistar rats were randomly divided into 6 groups (n = 3 in each group). For each rat, one eye was injured by alkali burn, the other one was taken as the normal control. Then all the corneas were surgically removed and the expression of laminin and fibronectin was observed with immunohistochemistry respectively at 7 hours, 1 day, 3 days, 7 days, 14 days and 28 days after alkali burn. RESULTS: Compared with that of the normal controls, the expression of laminin and fibronectin of the burned eyes was dramatically higher at 7 hours, reached peak at 14 days and decreased to the normal level at 28 days after alkali burn. CONCLUSIONS: In the process of wound healing after alkali burn, the expression of laminin and fibronectin increases dramatically, which suggests that laminin and fibronectin may participate in the process of corneal wound healing.