Knockdown of HDAC9 Inhibits Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Partially by Suppressing the MAPK Signaling Pathway.

Background: Histone deacetylase 9 (HDAC9) is a member of the HDAC gene family that plays essential roles in the organization of transcriptional regulation by catalyzing deacetylation of histone proteins. However, the effects of HDAC9 on osteonecrosis of femoral head (ONFH) have not been investigated. The present study aimed to reveal whether histone deacetylase 9 (HDAC9) regulated osteogenic differentiation. Methods: A lentiviral knockdown HDAC9 model was established in hBMSCs. Osteoblast-specific gene expression, such as Runx2, OCN was examined by qRT-PCR and Western blot, respectively. Though transcriptome sequencing and enrichment analysis, related signal pathways caused by down-regulation of HDAC9 were screened. The effect of HDAC9 on MAPK signaling pathway was determined by Western blot. Eventually, tert-Butylhydroquinone (tBHQ) was used to examine the effect of MAPK activation on osteogenesis in HDAC9 knockdown hBMSCs. Results: A lentiviral knockdown HDAC9 model was successfully established in hBMSCs. HDAC9 knockdown significantly inhibited osteoblast-specific gene expression, such as runt-related transcription factor 2 (Runx2), osteocalcin (OCN) and mineral deposition in vitro. Moreover, a total of 950 DEGs were identified in HDAC9-knockdown hBMSCs. We discovered that the MAPK signaling pathway might be related to this process by pathway enrichment analysis. HDAC9 knockdown significantly reduced the expression level of phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2). Finally, the decreased osteogenesis due to HDAC9 knockdown was partly rescued by a MAPK signaling pathway activator. Conclusion: Taken together, these results suggest that HDAC9 knockdown inhibits osteogenic differentiation of hBMSCs, partially through the MAPK signaling pathway. HDAC9 may serve as a potential target for the treatment of ONFH.

Comparison of the Clinical Outcomes of Full-Endoscopic Visualized Foraminoplasty and Discectomy Versus Microdiscectomy for Lumbar Disc Herniation.

OBJECTIVE: This retrospective case-control study aimed to evaluate and compare the clinical outcomes of full-endoscopic visualized foraminoplasty and discectomy (FEVFD) with microdiscectomy (MD) for lumbar disc herniation (LDH). METHODS: Data from 198 patients who presented with LDH between January 2016 and December 2017 treated by either FEVFD or MD were retrospectively analyzed. The inclusion criteria were single-level LDH, unilateral radiating leg pain with or without positive Lasegue's sign, and failure of standard conservative treatment for at least 12 weeks. The patients were categorized into an FEVFD group (n = 102) or an MD group (n = 96), according to the surgical procedure performed. Operative time, time in bed after surgery, postoperative hospitalization time, complications, and reoperations were recorded. Visual analog scales (VAS) for leg and back pain, Oswestry Disability index (ODI), 36-Item Short-Form Health Survey physical function (SF36-PF), and bodily pain (SF36-BP) scores were assessed and compared between the two groups. RESULTS: The demographic data and baseline characteristics of the two groups were not significantly different. Operative time for the FEVFD group (73.82 ± 20.73 min) was longer than that for the MD group (64.74 ± 17.37 min) (P = 0.003), and fluoroscopy time for the FEVFD group (1.71 ± 0.58s) was longer than that for the MD group (1.30 ± 0.33s) (P < 0.001). However, time in bed experienced in the FEVFD group (8.51 ± 2.10 h) was less than that in the MD group (9.24 ± 2.01 h) (P = 0.014), and postoperative hospitalization time experienced in the FEVFD group (2.89 ± 0.83d) was also shorter than that in the MD group (4.94 ± 1.35d) (P < 0.001). All patients completed 24 months of follow-up. Postoperative scores at each follow-up for the VAS for leg and back pain, ODI, SF36-PF, and SF36-BP all improved significantly for both groups, as compared to the preoperative data (P < 0.05). The mean preoperative and postoperative scores for the VAS for leg and back pain, ODI, SF36-PF, and SF36-BP were not significantly different between the two groups. According to the modified MacNab criteria, the outcomes of the procedures were rated as excellent or good by 92.16% and 93.75% of the patients in the FEVFD and MD groups, respectively. One patient suffered a nerve root injury during the discectomy, one patient suffered from a dural tear, and two patients suffered from a residual herniation in the FEVFD group. One patient in the MD group suffered from poor wound healing. Moreover, recurrence happened in two cases in the FEVFD group, and in one case in the MD group. CONCLUSION: FEVFD and MD are both reliable techniques for the treatment of symptomatic LDH. FEVFD resulted in a more rapid recovery and equivalent clinical outcomes after 24 months of follow-up.

New Concealed-Incision Extrapelvic Approach for Pubic Symphysis Diastasis and Parasymphyseal Fractures: Preliminary Results.

BACKGROUND: The Pfannenstiel approach, which provides good surgical exposure, has been used for the treatment of pubic symphysis diastasis and parasymphyseal fractures. However, it requires a medium-length incision and moderate soft-tissue dissection, resulting in potential damage to anatomical structures and inferior aesthetic outcomes. Here, we introduce a new concealed-incision extrapelvic approach for the internal fixation of pubic symphysis diastasis and parasymphyseal fractures. METHODS: We retrospectively reviewed the records of 8 patients with pubic symphysis diastasis and parasymphyseal fractures that had been treated via the concealed-incision extrapelvic approach (the "Fu-Liu" approach). All patients presented for treatment during the period from January 2017 to November 2017. Six of the 8 patients had anterior column fractures, 1 patient had a double-column fracture, and 1 patient had parasymphyseal fractures. Operative time, the amount of blood loss, and postoperative radiographic and computed tomography (CT) findings were recorded. The degree of fracture-healing, complications, function, and satisfaction with the skin incisions were also evaluated. RESULTS: All patients were followed for at least 21 months (range, 21 to 30 months). Postoperative radiographs and CT scans showed good positioning of plates and screws. The average time before surgery, operative time, and intraoperative blood loss (and standard deviation) were 7.8 ± 3.25 days, 41.9 ± 8.99 minutes, and 18.8 ± 7.8 mL, respectively. No complications (including internal fixation failure, vascular injury, nerve palsy, wound infection, and hernia) occurred in any of the patients, and all patients were satisfied with the appearance of the scar. CONCLUSIONS: We can effectively stabilize pubic symphysis diastasis and parasymphyseal fractures with use of the Fu-Liu approach, which can also enable retrograde anterior column screw placement. The Fu-Liu approach is simple, safe, and minimally invasive, and the aesthetic outcome is more acceptable than that associated with the Pfannenstiel approach. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Quantitative analysis by reversed-phase high-performance liquid chromatography and retinal neuroprotection after topical administration of moxonidine.

AIM: To determine moxonidine in aqueous humor and iris-ciliary body by reversed-phase high performance liquid chromatography (RP-HPLC), and to evaluate the retinal neuroprotective effect after topical administration with moxonidine in a high intraocular pressure (IOP) model. METHODS: The eyes of albino rabbits were administered topically and ipsilaterally with 0.2% moxonidine. A RP-HPLC method was employed for the identification and quantification of moxonidine between 2 and 480min, which presented in the aqueous humor and iris-ciliary body. Flash electroretinography (F-ERG) amplitude and superoxide dismutase (SOD) level were measured between day 1 and day 15 after topical administration with moxonidine in a rabbit model of high IOP. Histological and ultrastructural observation underwent to analyze the changes of retinal morphology, the inner retinal layers (IRL) thickness, and retinal ganglion cell (RGC) counting. RESULTS: Moxonidine was detectable between 2 and 480min after administration, and the peak concentration developed both in the two tissues at 30min, 0.51 µg/mL in aqueous humor and 1.03 µg/g in iris-ciliary body. In comparison to control, F-ERG b-wave amplitude in moxonidine eyes were significantly differences between day 3 and day 15 (P<0.01) in the high IOP model; SOD levels were significantly higher at all time-points (P<0.01) with a maximum level of 20.29 U/mgprot at day 15; and RGCs were significantly higher (P<0.05). CONCLUSION: Moxonidine is a viable neuroprotective agent with application to high IOP model. All layers of retina, including RGC layer, retinal nerve fiber layer and INL, are more preserved after moxonidine administration. SOD plays a neuroprotective role in ocular hypertension-mediated RGC death.

Blending of PLGA-PEG-PLGA for Improving the Erosion and Drug Release Profile of PCL Microspheres.

BACKGROUND: PCL has a long history as an industrialized biomaterial for preparing microspheres, but its hydrophobic property and slow degradation rate often cause drug degeneration, quite slow drug release rate and undesirable tri-phasic release profile. MATERIALS AND METHODS: In this study, we used the blending material of PLGA-PEG-PLGA and PCL to prepare microspheres. The microspheres degradation and drug release behaviors were evaluated through their molecular weight reduction rate, mass loss rate, morphology erosion and drug release profile. The hydrophilic PLGA-PEG-PLGA is expected to improve the degradation and drug release behaviors of PCL microspheres. RESULTS: Microspheres in blending materials exhibited faster erosion rates than pure PCL microspheres, forming holes much quickly on the particle's surface for the drug to diffuse out. A higher proportion of PLGA-PEG-PLGA caused faster degradation and erosion rates. The blending microspheres showed much faster drug release rates than pure PCL microspheres. CONCLUSION: With blending of 25wt% PLGA-PEG-PLGA, the release rate of microspheres speeded up significantly, while, with a further increase of PLGA-PEG-PLGA proportion (50%, 75%, 100%), it accelerated a little. The microspheres with PCL/PLGA-PEG-PLGA of 1/1 exhibited a linear-like drug release profile. The results could be a guideline for preparing microspheres based on blending materials to obtain a desirable release.

Age-related Changes in the Global DNA Methylation Profile of Oligodendrocyte Progenitor Cells Derived from Rat Spinal Cords.

Demyelination of axons plays an important role in the pathology of many spinal cord diseases and injuries. Remyelination in demyelinated lesions is primarily performed by oligodendrocyte progenitor cells (OPCs), which generate oligodendrocytes in the developing and mature central nervous system. The efficiency of remyelination decreases with age. Many reports suggest that this decline in remyelination results from impaired OPC recruitment and differentiation during aging. Of the various molecular mechanisms involved in aging, changes in epigenetic modifications have received particular attention. Global DNA methylation is a major epigenetic modification that plays important roles in cellular senescence and organismal aging. Thus, we aimed to evaluate the dynamic changes in the global DNA methylation profiles of OPCs derived from rat spinal cords during the aging process. We separated and cultured OPCs from the spinal cords of neonatal, 4-month-old, and 16-month-old rats and investigated the age-related alterations of genomic DNA methylation levels by using quantitative colorimetric analysis. To determine the potential cause of dynamic changes in global DNA methylation, we further analyzed the activity of DNA methyltransferases (DNMTs) and the expression of DNMT1, DNMT3a, DNMT3b, TET1, TET2, TET3, MBD2, and MeCP2 in the OPCs from each group. Our results showed the genomic DNA methylation level and the activity of DNMTs from OPCs derived from rat spinal cords decreased gradually during aging, and OPCs from 16-month-old rats were characterized by global hypomethylation. During OPC aging, the mRNA and protein expression levels of DNMT3a, DNMT3b, and MeCP2 were significantly elevated; those of DNMT1 were significantly down-regulated; and no significant changes were observed in those for TET1, TET2, TET3, or MBD2. Our results indicated that global DNA hypomethylation in aged OPCs is correlated with DNMT1 downregulation. Together, these data provide important evidence for partly elucidating the mechanism of age-related impaired OPC recruitment and differentiation and assist in the development of new treatments for promoting efficient remyelination.

MiR-708 promotes steroid-induced osteonecrosis of femoral head, suppresses osteogenic differentiation by targeting SMAD3.

Steroid-induced osteonecrosis of femoral head (ONFH) is a serious complication of glucocorticoid (GC) use. We investigated the differential expression of miRs in the mesenchymal stem cells (MSCs) of patients with ONFH, and aimed to explain the relationship between GC use and the development of MSC dysfunction in ONFH. Cells were collected from bone marrow of patients with ONFH. Samples were assigned to either GCs Group or Control Group at 1:1 matched with control. We then used miRNA microarray analysis and real-time PCR to identify the differentially expressed miRs. We also induced normal MSCs with GCs to verify the differential expression above. Subsequently, we selected some of the miRs for further studies, including miRNA target and pathway prediction, and functional analysis. We discovered that miR-708 was upregulated in ONFH patients and GC-treated MSCs. SMAD3 was identified as a direct target gene of miR-708, and functional analysis demonstrated that miR-708 could markedly suppress osteogenic differentiation and adipogenesis differentiation of MSCs. Inhibition of miR-708 rescued the suppressive effect of GC on osteonecrosis. Therefore, we determined that GC use resulted in overexpression of miR-708 in MSCs, and thus, targeting miR-708 may serve as a novel therapeutic biomarker for the prevention and treatment of ONFH.

Association Between Lumbar Disc Degeneration and Propionibacterium acnes Infection: Clinical Research and Preliminary Exploration of Animal Experiment.

STUDY DESIGN: Clinical research and animal experiment. OBJECTIVE: To investigate whether lumbar disc degeneration is associated with Propionibacterium acnes (P acnes) infection. SUMMARY OF BACKGROUND DATA: The hypothesis that herniated discs may be infected with P acnes by way of bacteremia is remarkable. This may bring a tremendous change in treatment of lumbar disc herniation (LDH). However, this hypothesis is still controversial. Since P acnes isolated may be related to contamination. METHODS: Nucleus pulposus from 22 patients (30 discs) with lumbar disc herniation was collected during discectomy, following aerobic and anaerobic cultures for 10 days.Twenty-four rabbits were divided into four groups. After L3-L6 being exposed, an incision was made into the three discs in groups A and B. While in groups C and D, two random segments were operated. Six weeks later, 0.05 mL of 5 × 10 CFU/mL P acnes was inoculated into operated discs in group A and sterile physiological saline in group B. In group C, 0.2 mL of 5 × 10 CFU/mL P acnes was injected through ear vein. Sterile saline was used in group D. Six weeks later, MRI was performed. Then, nucleus pulposus and paraspinal muscles were harvested for aerobic and anaerobic cultures. RESULTS: Clinical research: Anaerobic cultures were positive in three cases: two coagulase-negative staphylococci, one particles chain bacterium. No P acnes was found. Staphylococcus epidermidis was isolated in one aerobic culture.Animal experiment: P acnes was found in 11 out of 18 (61%) discs in group A. There was no P acnes found in the other three groups. CONCLUSION: Degenerated discs were suitable for P acnes growth. This research did not find the evidence of the symptomatic degenerated lumbar discs infected with P acnes or that P acnes could infect the degenerated lumbar discs by way of bacteremia. LEVEL OF EVIDENCE: N/A.

Comparison of the outcomes of cannulated screws vs. modified tension band wiring fixation techniques in the management of mildly displaced patellar fractures.

BACKGROUND: K wire fixation with tension band wiring has conventionally been used for the open reduction and internal fixation of the patella. However, it suffers from distinct disadvantages such as implant irritation, need for open reduction, incidence of palpable implants, and need for subsequent implant removal. A smaller incision with percutaneous fixation may be an alternative to this established conventional technique. Thus, the purpose of this trial was to compare the treatment outcomes of patients with mildly displaced patellar fractures treated with closed reduction and percutaneous cannulated screw fixation (CRCF) as compared to open reduction and tension band wiring fixation (ORTF). Specifically, we aimed to determine whether cannulated screw fixation was associated with improved clinical outcomes at 12 months as measured using the Lysholm score, pain scores, degree of flexion, range of motion, time to radiographic union, radiographic outcomes, and complication rates. METHODS: Sixty-three patients with transverse patellar fractures displaced less than 8 mm were included in this prospective, randomized, controlled trial, with 52 patients in the final data analysis. Thirty-two patients were operatively treated by CRCF with either two or three cannulated screws. Thirty-one patients were operatively treated by conventional ORTF using the modified tension band technique. At postoperative intervals of 3, 6, and 12 months, knee function was evaluated using the Lysholm score, pain was assessed using the visual analog scale (VAS) score, and active knee extensions and flexion were measured in degrees by goniometry. RESULTS: The CRCF group had average Lysholm scores of 84.4 ± 5.8, 86.7 ± 6.4, and 93.2 ± 5.3 after 3, 6, and 12 months, respectively, which were significantly greater than those of the ORTF group (79.0 ± 5.3, p = 0.001; 81.5 ± 4.6, p = 0.002; and 89.8 ± 6.2, p = 0.039, respectively). Lower pain and squatting scores were the main reasons for the poorer Lysholm scores in the ORTF group. The VAS scores showed that the CRCF group had lower pain scores and better flexion and total range of motion (ROM) compared with the ORTF group after 3 and 6 months, although both groups had similar outcomes after 12 months. The mean fracture healing time of 2.65 months was similar in the CRCF groups (2.77 months; p = 0.440). Complication rates were 3/26 (11.5 %) in the CRCF group and 14/26 (53.4 %) in the ORTF group. Two patients in the CRCF group and eight patients in the ORTF group experienced skin irritation. In addition, two (7.7 %) patients in the CRCF group and 11 (42.3 %) patients in the ORTF group required implant removal because of symptoms due to the presence of the implants. CONCLUSION: Surgical treatment of mild displaced (less than 8 mm) transverse patellar fractures by the CRCF technique provides satisfactory clinical results and excellent knee function, with little pain and a low incidence of complications at early follow-up (up to 6 months). These results suggest that the CRCF technique may be a superior alternative to conventional ORTF. Registration Trial (Chinese Clinical Trial Register): Current Controlled Trials ChiCTR-PRCH-14005017, registration dates 2014-06-14.

Controlled chondrogenesis from adipose-derived stem cells by recombinant transforming growth factor-ß3 fusion protein in peptide scaffolds.

Adipose-derived stem cells (ADSCs) are promising for cartilage repair due to their easy accessibility and chondrogenic potential. Although chondrogenesis of transforming growth factor-ß (TGF-ß) mediated mesenchymal stem cells (MSCs) is well established in vitro, clinical tissue engineering requires effective and controlled delivery of TGF-ß in vivo. In this work, a self-assembled peptide scaffold was employed to construct cartilages in vivo through the chondrogenesis from ADSCs controlled by recombinant fusion protein LAP-MMP-mTGF-ß3 that was transfected by lentiviral vectors. During this course, the addition of matrix metalloproteinases (MMPs) can trigger the release of mTGF-ß3 from the recombinant fusion protein of LAP-MMP-mTGF-ß3 in the combined scaffolds, thus stimulating the differentiation of ADSCs into chondrogenesis. The specific expression of cartilage genes was analyzed by real-time polymerase chain reaction and Western blot. The expression of chondrocytic markers was obviously upregulated to a higher level compared to the one by commonly used TGF-ß3 alone. After 3 weeks of in vitro culturing, the hybrids with differentiated chondrogenesis were then injected subcutaneously into nude mice and retrieved after 4 weeks of culturing in vivo. Histological analysis also confirmed that the recombinant fusion protein was more effective for the formation of cartilage matrix than the cases either with TGF-ß3 alone or without LAP-MMP-mTGF-ß3 (P<0.05). This study demonstrates that controlled local delivery of the LAP-MMP-mTGF-ß3 constructs can accelerate differentiation of ADSCs into the cartilage in vivo, which indicates the great potential of this hybrid in rapid therapy of osteoarthritis.

Minimally invasive plate osteosynthesis with a locking compression plate is superior to open reduction and internal fixation in the management of the proximal humerus fractures.

BACKGROUND: The use of minimally invasive plate osteosynthesis (MIPO) via anterolateral deltoid splitting has good outcomes in the management of proximal humerus fractures. While using this approach has several advantages, including minimal soft tissue disruption, preservation of natural biology and minimal blood loss, there is an increased risk for axillary nerve damage. This study compared the advantages and clinical and radiological outcomes of MIPO or open reduction and internal fixation (ORIF) in patients with proximal humerus fractures. METHODS: A matched-pair analysis was performed, and patient groups were matched according to age (±3 years), sex and fracture type. Forty-three pairs of patients (average age: MIPO, 63 and ORIF, 61) with a minimum follow-up of 12 months were enrolled in the study group. The patients were investigated radiographically and clinically using the Constant score. RESULTS: The MIPO technique required less surgery time and caused less blood loss compared to ORIF (p < 0.01). In addition, MIPO required a smaller incision, resulted in less scarring, and was cosmetically more appealing and acceptable to female patients than ORIF. Following MIPO, patients had better functional results at 3 and 6 months, with better outcomes, less pain, higher satisfaction in activities of daily living, and a higher range of motion when compared to ORIF (p < 0.05). Fracture configuration, according to the AO/ASIF(Association for the Study of Internal Fixation) fracture classification, did not significantly influence the functional results. The complication rate was comparable between both groups. CONCLUSION: The use of MIPO with a locking compression plate in the management of proximal humerus fractures is a safe and superior option compared to ORIF.

[Cannulated lag screw combined with lateral supporting plate for treatment of Hoffa fracture of Letenneur type I and type III].

OBJECTIVE: To investigate the effectiveness of cannulated lag screws combined with lateral supporting plates in the treatment of Hoffa fracture of Letenneur type I and type III. METHODS: Between May 2004 and April 2011, 11 patients with Hoffa fracture of Letenneur type I and type III were treated, including 6 males and 5 females with an average age of 36 years (range, 25-47 years). Factures were caused by traffic accident in 8 cases, by falling in 2 cases, and by the other in 1 case. Fracture involved the left knee in 7 patients and the right knee in 4 patients. According Letenneur's classification criteria, there were 7 type I fractures (6 lateral condyle fractures and 1 medial condyle fracture) and 4 type III fractures (3 lateral condyle fractures and 1 medial condyle fracture). Of 11 fractures, 9 were fresh fractures and 2 were old fractures. Two 6.5 mm cannulated lag screws combined with lateral supporting plates were used to fix fractures by anterolateral or anteromedial incision. RESULTS: All incisions achieved primary healing with no early complication. All patients were followed up 12-26 months (mean, 15 months). X-ray films showed bone healing with an average healing time of 15 weeks (range, 10-18 weeks). No loosening or breaking of internal fixator was observed; the removal time of internal fixation was 9-15 months (mean, 12 months). Accoding to Letenneur's functional assessment system, the results were excellent in 7 cases, good in 3 cases, and poor in 1 case at last follow-up. CONCLUSION: Cannulated lag screws combined with lateral supporting plates fixation is effective in treatment of Hoffa fracture of Letenneur type I and type III with a high union rate; anterolateral or anteromedial approach is the first choice for Hoffa fracture of type I and type III, especially for complicating by tibial plateau fracture or patella fracture.

Surgical technique: repair of forefoot skin and soft tissue defects using a lateral tarsal flap with a reverse dorsalis pedis artery pedicle: a retrospective study of 11 patients.

BACKGROUND: Various authors have proposed flaps to reconstruct traumatic forefoot skin and soft tissue defects, especially with exposure of tendon and/or bone although which is best for particular circumstances is unclear. DESCRIPTION OF TECHNIQUE: The indications for the technique were a forefoot defect area of no more than 8-cm × 8-cm and a well-preserved lateral tarsal (LT) donor site. The injured tendons were repaired using tendon grafts. The free dorsalis pedis flap was outlined by centering it on the cutaneous branch of the LT artery and tailoring it to the size of the wound, allowing 0.5-cm margins in length and width. The flap was rotated around the plantar perforating branch of the dorsalis pedis artery (DPA) to cover the forefoot defect. The lateral dorsalis pedis cutaneous nerve was anastomosed with the recipient plantar nerve stump. The donor site was covered with an inguinal, full-thickness skin graft. PATIENTS AND METHODS: Traumatic forefoot skin and soft tissue defects with exposure of the tendon and/or bone involving 11 feet in 11 patients (mean age, 32 years) were covered using a LT flap with a reversed DPA pedicle. Three patients with forefoot defects underwent emergency repair within 8 hours of injury, whereas eight patients required delayed repair. All patients were followed up for at least 6 months (mean, 13 months; range, 6-24 months). RESULTS: All flaps survived uneventfully, except for two that had superficial marginal necrosis or severe venous insufficiency. All skin grafts covering the donor sites survived and all wounds healed. None of the patients had restricted standing or walking at followups. The two-point discrimination was 4 mm to 10 mm at 6 months postoperative. The mean hallux-metatarsophalangeal-interphalangeal scale score was 93 points (range, 87-98 points). CONCLUSIONS: Our observations suggest the LT flap with a reversed DPA pedicle is a reasonable option for repair of traumatic forefoot skin and soft tissue defects with exposure of tendon and/or bone but a well-preserved LT donor site and is associated with minimal morbidity.

Comparison of endoscopic surgery and open surgery for gluteal muscle contracture.

PURPOSE: To compare the clinical effects of endoscopic surgeries with traditional open surgeries in the treatment of gluteal muscle contracture and discuss their indications and value. METHODS: In this retrospective study, 50 patients received traditional open surgeries and 52 received endoscopic surgeries. The 2 groups were compared in terms of surgery duration, incision lengths, postsurgical pain, complications, off-bed activity times, hospitalization duration, clinical outcome, and 1-year recurrence rates. RESULT: The endoscopic surgery group was significantly superior to the open surgery group in regard to incision length, postsurgical pain, off-bed activity time, hospitalization duration, and patient cosmetic satisfaction. Differences were not statistically significant for the surgery duration, complications, clinical outcome, or the 1-year recurrence rate. All the endoscopic surgery group patients stated that they would choose endoscopic surgery again. CONCLUSIONS: The endoscopic release of gluteal muscle contracture is safe and reliable, with the advantages of less trauma and pain, shorter operative time, earlier rehabilitation, and return of functional activities. Its application, though, should be carefully controlled based on the indications. It is applicable to degree I and II patients, but may be used only very cautiously in degree III patients. LEVEL OF EVIDENCE: Level III.

Open reduction and bioabsorbable pin fixation for late presenting irreducible supracondylar humeral fracture in children.

The objective was to evaluate the availability and efficacy of internal fixation with absorbable poly-D: ,L: -lactic acid (PDLLA) pins for the treatment of late presenting irreducible Gartland type III supracondylar fracture of the humerus in children. Fifty-six cases of late presenting irreducible Gartland type III supracondylar fracture of the humerus in children were treated by open reduction and bioabsorbable PDLLA pin fixation from March 2005 to March 2008. The outcome of treatment was evaluated by the Mayo Elbow Performance Score (MEPS) and the criteria of Flynn. Fifty-six patients were followed up from 24 to 36 months (mean: 22 months). No displacement of bone fracture occurred, and all fractures healed within a normal time without wound infection; there were no cases of Volkmann's ischaemic contracture, myositis ossificans or iatrogenic injury of the ulnar nerve. No residual vascular deficits or iatrogenic nerve injury were noted; cubitus varus deformity occurred in one case. There were 49 excellent, four good and three fair results according to the MEPS; the rate of excellent and good outcome was 94.6%. All children but one had excellent cosmetic results according to the criteria of Flynn. All of the children and their parents stated that they would choose this treatment again. Treatment of late presenting irreducible Gartland type III supracondylar fracture of the humerus in children with bioabsorbable PDLLA pins provides sufficient stability and satisfactory efficacy. The absorbable implant has become popular for its avoidance of a second operation to remove the internal fixation, and the degree of patient satisfaction is high.

Mobilization efficiency of granulocyte colony-stimulating factor and stem cell factor to bone marrow mononuclear cells and mechanisms.

The mobilization efficiency of granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) to bone marrow mononuclear cells (MNCs) in mice was observed, and the changes of CXCL12/CXCR4 signal were detected in order to find out the mobilization mechanism of stem cells. Kunming mice were randomly divided into two groups. The mice in treatment group were subjected to subcutaneous injection of G-CSF at a dose of 100 microg/kg and SCF at a dose of 25 microg/kg every day for 5 days, and those in control group were given isodose physiological saline. The MNCs were separated, counted and cultured, and the colony-forming unit-fibroblast (CFU-F) was evaluated. CD34+CXCR4+ MNCs were sorted by flow cytometry. The expression of CXCL12 protein in bone marrow extracellular fluid was detected by ELISA, and that of CXCL12 mRNA in bone marrow was measured by RT-PCR. The results showed that the counts of MNCs in peripheral blood and bone marrow were increased after administration of G-CSF/SCF (P<0.01). The factors had a dramatic effect on the expansion capability of CFU-F (P<0.05). Flow cytometric of bone marrow MNCs surface markers revealed that CD34+CXCR4+ cells accounted for 44.6%+/-8.7% of the total CD34+ MNCs. Moreover, G-CSF/SCF treatment induced a decrease in bone marrow CXCL12 mRNA that closely mirrored the fall in CXCL12 protein. In this study, it is evidenced that G-CSF/SCF can effectively induce MNCs mobilization by disrupting the balance of CXCL12/CXCR4 signaling pathway in the bone marrow and down-regulating the interaction of CXCL12/CXCR4.

Decreased in the number and function of circulation endothelial progenitor cells in patients with avascular necrosis of the femoral head.

INTRODUCTION: Once non-traumatic avascular necrosis of the femoral head (ANFH) happened, vascular impairment and feeble collateral circulation are followed by poor outcomes. Circulating endothelial progenitor cells (EPCs) may substantially contribute to vascular homeostasis such as vascular repair and new blood vessel growth. We investigated whether abnormalities in EPCs levels and functions are present in ANFH patients. METHODS: 54 ANFH patients were enrolled, including steroid-induced (n=21), alcohol-induced (n=15) and idiopathic ANFH (n=18), and 30 healthy subjects as control (HC). The numbers of circulation EPCs were determined by fluorescence-activated cell-sorting (FACS) analysis. EPCs cultured from peripheral blood mononuclear cells on fibronectin to induce the expression of receptors for acetylated low-density lipoprotein and ulex-lectin. EPCs colony-forming units (CFUs) were observed from 54 patients and 30 healthy controls. Migratory capacity to chemo-attractants (vascular endothelial growth factor) cellular senescence levels and in vitro angiogenesis ability were assessed in age-matched subjects (n=10 per groups). RESULTS: Mean numbers of circulating EPC were 1460+/-265 cells/ml in HC, 545+/-177 in ANFH, (P<0.001). Mean numbers of CFUs were 26.2+/-6.2 in HC, 19.6+/-7.7 in ANFH,(P<0.001). Although there were not significant differences in circulating EPC and CFUs among the steroid-induced, alcohol-induced or idiopathic three groups, all these risk factors contributed to the decreased circulating EPCs numbers and CFUs. In addition, EPCs from ANFH patients showed reduced migratory capacity and increased cellular senescence compared with EPCs from normal subjects, furthermore the ability of angiogenesis in vitro was also impaired. CONCLUSION: Circulating endothelial progenitor cells (EPCs) numbers and functions are reduced in ANFH patients, suggesting that risk factors of ANFH may alter EPCs biology in angiogenesis and vascular repair.

Mobilization Efficiency of Granulocyte Colony-stimulating Factor and Stem Cell Factor to Bone Marrow Mononuclear Cells and Mechanisms / 华中科技大学学报（医学）（英德文版）

The mobilization efficiency of granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) to bone marrow mononuclear cells (MNCs) in mice was observed,and the changes of CXCL12/CXCR4 signal were detected in order to find out the mobilization mechanism of stem cells.Kunming mice were randomly divided into two groups.The mice in treatment group were subjected to subcutaneous injection of G-CSF at a dose of 100 μg/kg and SCF at a dose of 25 μg/kg every day for 5 days,and those in control group were given isodose physiological saline.The MNCs were separated,counted and cultured,and the colony-forming unit-fibroblast (CFU-F) was evaluated.CD34+CXCR4+ MNCs were sorted by flow cytometry.The expression of CXCL12 protein in bone marrow extracellular fluid was detected by ELISA,and that of CXCL12 mRNA in bone marrow was measured by RT-PCR.The results showed that the counts of MNCs in peripheral blood and bone marrow were increased after administration of G-CSF/SCF (P＜0.01).The factors had a dramatic effect on the expansion capability of CFU-F (P＜0.05).Flow cytometric of bone marrow MNCs surface markers revealed that CD34+CXCR4+ cells accounted for 44.6%±8.7% of the total CD34+ MNCs.Moreover,G-CSF/SCF treatment induced a decrease in bone marrowCXCL12 mRNA that closely mirrored the fall in CXCL12 protein.In this study,it is evidenced that G-CSF/SCF can effectively induce MNCs mobilization by disrupting the balance of CXCL12/CXCR4 signaling pathway in the bone marrow and down-regulating the interaction of CXCL12/CXCR4.

Bone regeneration with active angiogenesis by basic fibroblast growth factor gene transfected mesenchymal stem cells seeded on porous beta-TCP ceramic scaffolds.

Large segmental bone defect repair remains a clinical and scientific challenge with increasing interest focused on combining gene transfer with tissue engineering techniques. Basic fibroblast growth factor (bFGF) is one of the most prominent osteogenic growth factors that has the potential to accelerate bone healing by promoting the proliferation and differentiation of mesenchymal stem cells (MSCs) and the regeneration of capillary vasculature. However, the short biological half-lives of growth factors may impose severe restraints on their clinical usefulness. Gene-based delivery systems provide a better way of achieving a sustained high concentration of growth factors locally in the defect and delivering a more biologically active product than that achieved by exogenous application of recombinant proteins. The objective of this experimental study was to investigate whether the bFGF gene modified MSCs could enhance the repair of large segmental bone defects. The pcDNA3-bFGF gene transfected MSCs were seeded on biodegradable porous beta tricalcium phosphate (beta-TCP) ceramics and allografted into the 15 mm critical-sized segmental bone defects in the radius of 18 New Zealand White rabbits. The pcDNA3 vector gene transfected MSCs were taken as the control. The follow-up times were 2, 4, 6, 8, 10 and 12 weeks. Scanning electron microscopic, roentgenographic, histologic and immunohistological studies were used to assess angiogenesis and bone regeneration. In vitro, the proliferation and differentiation of bFGF gene transfected MSCs were more active than that of the control groups. In vivo, significantly more new bone formation accompanied by abundant active capillary regeneration was observed in pores of the ceramics loaded with bFGF gene transfected MSCs, compared with control groups. Transfer of gene encoding bFGF to MSCs increases their osteogenic properties by enhancing capillary regeneration, thus providing a rich blood supply for new bone formation. This new bFGF gene enhanced tissue engineering strategy could be of potential benefit to accelerate bone healing, especially in defects caused by atrophic nonunion and avascular necrosis of the femoral head.

Effects of TGF beta1 autocrine blockage on osteosarcoma cells.

OBJECTIVE: To evaluate the effects of transforming growth factor beta1 (TGFbeta1) autocrine blockage on proliferation activity and drug sensitivity of osteosarcoma. METHODS; Northern blot, MTT determination, and 3H thymidine incorporation were used to investigate the effects of antisense TGF beta1 gene on osteosarcoma. RESULTS: The proliferation of osteosarcoma cells transfected by antisense TGF beta1 gene was suppressed markedly, and adriamycin sensitivity was significantly increased. CONCLUSION: Blockage of osteosarcoma cells TGF beta1 autocrine loop inhibits cell proliferation and enhances chemotherapy sensitivity.