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Objective:To study the effects of enoxaparin on osteogenic differentiation of bone marrow stem cells (BMSCs) and the exosomes derived from BMSCs.Methods:After the BMSCs from 4-week old male SD rats were cultured, their surface antigen and multilineage differentiation potentials were identified. Subsequently, the BMSCs were incubated with osteogenic differentiation medium containing 10 IU/mL enoxaparin for 14 days. After the exosomes derived from BMSCs (BMSC-Exos) were extracted by the kit method, their structure was observed by transmission electron microscopy and their surface antigen CD63 detected by Western blot. Alizarin red staining was used to analyze the osteogenic differentiation of BMSCs. Osteogenic proteins including OCN and BMP-2 in BMSCs and exosomes were detected.Results:The spindle-shaped BMSCs isolated and cultured showed a uniform spiral pattern. They expressed highly the surface markers CD29 and CD44 but lowly CD34 and CD45, indicating that the majority of the cells were BMSCs. BMSC-Exos, in an oval shape with a diameter of about 30 to 80 nm, expressed CD63. Alizarin red staining showed that the number of mineralized nodules in the enoxaparin treatment group was significantly lower than that in the control group. Western blot analysis indicated that enoxaparin inhibited the expression of osteocalcin (OCN) and BMP-2 in BMSCs. ELISA results showed that the protein levels of OCN(48.81 ng/mL ± 8.23 ng/mL) and BMP-2 (311.45 pg/mL ± 27.59 pg/mL) in the BMSCs treated with enoxaparin were significantly lower than those of OCN (80.43 ng/mL ± 10.74 ng/mL) and BMP-2 (399.23 pg/mL ± 32.25 pg/mL) in the control BMSCs ( P<0.05). The contents of OCN (1.45 ng/mL±0.15 ng/mL) and BMP-2 (18.47 pg/mL ± 0.54 pg/mL) in the exosomes from BMSCs treated with enoxaparin were significantly higher than those of OCN (1.00 ng/mL ± 0.12 ng/mL) and BMP-2 (9.07 ng/mL ± 0.36 pg/mL) in the exosomes from control BMSCs ( P<0.05). Conclusions:Enoxaparin may inhibit the osteogenic differentiation of BMSCs. The mechanism of this might be related to its direct inhibition against the expression of OCN and BMP-2 in BMSCs and its indirect reduction of OCN and BMP-2 in BMSCs through the exhaust of the above proteins in the form of exosomes.
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Objective To explore the clinical effect of training assisted by a lower limb rehabilitation robot on the bladder and intestinal function of paraplegic spinal cord injury survivors. Methods Thirty-eight paraplegic patients with spinal cord injury were divided according to their admission order into an experimental group ( n=19) and a control group (n=19). Both groups were given conventional rehabilitation training, while the experimental group was additionally provided with robot-assisted lower limb training in three stages:adaptation, training and con-solidation. It lasted 30 minutes daily, 5 days per week for 12 weeks. Before and after the training, an urodynamics examination system was used to evaluate the maximum urine flow, bladder capacity, residual urine volume, bladder pressure and detrusor pressure. Colon transit time, mean rectal pressure and intestinal function were measured using the colon transit test, a mean rectal pressure test, and the Functional Independence Measure ( FIM) scale respective-ly. Results The average bladder volume, maximum urine flow rate, average urine flow rate, detrusor pressure, bladder compliance, average rectal pressure and intestinal FIM score of the robot training group after training were all significantly better than before the training, as were the average residual urine volume and colon transit time. After the training, the average bladder volume, maximum urine flow rate, average urine flow rate, detrusor pressure, bladder compliance and average rectal pressure of the robot training group were all significantly higher than those of the control group, while the average residual urine volume and colon transit time were significantly smaller. Then, 32％ of the patients in the experimental group achieved no less than 6 points for their average FIM score, significantly higher than in the control group. Conclusion Robot-assisted lower limb training combined with comprehensive rehabilitation training can effectively improve the bladder and intestinal function of paraplegic patients after a spinal cord injury.
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Objective To preliminarily analyze the formation and significance of stem cell niche in the hair follicle bulge,based on the morphological structure of murine vibrissa follicles and localization of proliferative region of hair follicle stem cells at different developmental stages.Methods Vibrissa follicles of healthy,specific pathogen-free (SPF) outbred KM mice at different developmental stages were labelled with 5-ethynyl-2-deoxyuridine (EdU) as a marker of proliferating stem cells by in vivo local injection.After 24 hours,the complete vibrissa follicles were obtained by surgical microanatomy,and then subjected to OCT embedding,cryostat sectioning and hematoxylin-eosin (HE) staining.Sections labelled with EdU were subjected to immunofluorescence staining to determine the active proliferative region of hair follicle cells.Hair follicles were subjected to immunofluorescence staining with anti-stem cell transcription factor (Sox2) antibody.Results The morphology of hair follicles changed during the development,and mature hair follicles could be clearly observed in the hair follicle bulge.On day 1 to day 15 after birth,the active proliferative region of murine hair follicle cells was gradually transferred from the dermal papillae to the inner and outer root sheaths in the hair follicle bulb.Meanwhile,along with the inner and outer root sheaths,the actively proliferating hair follicle cells migrated upward to the middle and upper parts of hair follicles and formed the hair follicle bulge,in which the stem cell transcription factor Sox2 was abundantly expressed in cells.Conclusions Progenitor cells in the stem cell niche of the hair follicle bulge are derived from the dermal papillae of hair follicles.After the formation of the hair follicle bulge,cells start to abundantly express Sox2 and enter into the resting state.
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Objective To evaluate the clinical and radiologic outcomes of minimally invasive plate percutaneous osteosynthesis (MIPPO) for distal clavicle fractures of Neer type Ⅱ. Methods From May 2015 to May 2016, 13 patients with distal clavicle fracture of Neer type Ⅱwere treated with MIPPO using an anatomic locking plate. Anteroposterior plain X-ray images of the clavicle were taken at intervals until union was observed. Fracture healing time and complications were recorded, and shoulder joint functions evaluated using Constant and University of California at Los Angeles (UCLA) scores. Results The average operating time was 60.9 minutes (range, from 48 to 83 minutes), average intraoperative blood loss 24.7 mL (range, from 18 to 30 mL), and average union time for the patients was 13.3 weeks (range, from 11 to 15 weeks). No delayed union or nonunion occurred. There were no major complications, like infection, plate breakage, or neurovascular injury. Only 2 patients felt local incision numbness. All the patients were satisfactory with their shoulder func-tions. At the final follow-ups, the mean Constant score was 93.2 points (from 87 to 95 points) and the mean UCLA score 30.6 points (from 28 to 34 points). Conclusion In the treatment of distal clavicle fractures of Neer type Ⅱ, minimally invasive plate percutaneous osteosynthesis can not only achieve good clinical and radi-ologic outcomes but also be advantageous over conventional open reduction in aspects of smaller invasion and better cosmetic effect.
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Objective To optimize the culture medium for scarless wound healing in outbred fetal Kunming (KM) mice,and to locate and preliminarily analyze the region with active cell proliferation.Methods After 6 pregnant mice were sacrificed at the gestational age of 15 days,a total of 180 skin grafts were obtained from the back of 60 fetal mice,and wounds on the skin grafts were made uniformly with minimally invasive sterile device.Then,these skin grafts with wounds were randomly and equally divided into 6 groups to be treated with high-glucose Dulbecco's modified Eagle's medium (DMEM),DMEM + 5％ fetal bovine serum (FBS),DMEM + 10％ FBS,low-glucose Eagle's minimum essential medium (MEM),MEM + 5％ FBS,and MEM + 10％ FBS for 3 days,respectively.Then,these skin grafts were embedded in optimal cutting temperature (OCT) compound,and subjected to hematoxylin and eosin (HE) staining to select the optimal culture condition for wound healing.Under the optimal culture condition,the region with active cell proliferation was located by labeling and tracking cutaneous stem cells with 5-ethynyl-2-deoxyuridine (EdU),and activator protein-1 (AP-1) expression was detected by immunofluorescent staining.Results After 3-day cultivation,wound healing rates significantly differed among the DMEM group,DMEM + 5％ FBS group,DMEM + 10％ FBS group,MEM group,MEM + 5％ FBS group and MEM + 10％ FBS group (0,3.33％,6.67％,3.33％,46.67％ and 26.67％,respectively,x2 =41.39,P ＜ 0.05).Additionally,the MEM + 5％ FBS group showed a significantly higher wound healing rate than the other groups except the MEM + 10％ FBS group (all P ＜ 0.01).Logistic regression analysis showed that the type of basal medium (MEM:OR =11.717,95％ CI:3.274-41.934,P ＜ 0.001) and FBS concentrations (5％ FBS:OR =24.625,95％ CI:3.027-200.299,P =0.003;10％ FBS:OR =13.449,95％ CI:1.618-111.813,P =0.016) were factors influencing fetal wound healing.Under the culture condition of 5％ FBS + MEM,the dermis and epidermis healed well without epidermal thickening.The cutaneous stem cell labeling technique showed that the papillary dermis and basal layer of the epidermis were the two major regions with active cell proliferation in the wound of fetal mouse skin.Moreover,AP-1 was expressed abundantly in these two regions as well.Conclusions The culture condition of 5％ FBS + MEM is considered to be optimal for in vitro wound healing in fetal mice.The papillary dermis and basal layer of the epidermis are two major regions with active cell proliferation during wound healing in fetal mouse skin,and play important roles in wound healing process.
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Objective To compare the clinical outcomes of bone transport technique versus induced membrane technique for treatment of infected bone nonunion.Methods A retrospective study was made on 46 cases of infected bone nonunion admitted from April 2011 to December 2014.They were 33 males and 13 females,aged from 12 to 68 years (mean,45.3 years),involving 40 tibial and 6 femoral fractures.Bone transport technique was used in 37 cases while 9 cases were treated with induced membrane technique.Fracture union and complications were recorded.The function of the affected limb was evaluated at the final follow-up using Johner-Wruhs criteria.Results The 46 cases were followed up for 9 to 18 months (mean,14 months).In the bone transport group,bone union was achieved after a mean duration of 6.5 months (range,from 4 to 11 months) without complications like wound infection or neurovascular lesions.In the induced membrane group,bone union was achieved after a mean duration of 5.8 months (range,from 4 to 11 months) and there was one case of re-infection.According to the Johner-Wruhs score system,34 cases were rated as excellent,9 as good and 3 as fair,with a total excellent and good rate of 93.5％.Bone transport technique led to 28 excellent,4 good and one fair cases in the patients with tibial nonunion,and one excellent,2 good and one fair cases in the patients with femoral nonunion.Induced membrane technique led to 3 excellent,3 good and one fair cases in the patients with tibial nonunion,and 2 excellent cases in the patients with femoral nonunion.Conclusions Both bone transport technique and induced membrane technique are effective for infected bone nonunion.Bone transport technique may be preferred for tibial infected nonunion while induced membrane technique preferred for femoral infected nonunion.
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Objective To explore the clinical efficacy of locking plate in the treatment of postoperative nonunion of limb long bone fracture.Methods We treated 53 patients whose limb long bone fracture failed to unite after operation with locking plate between April 2011 and November 2014.They were 38 men and 15 women,aged from 20 to 69 (average,39.7).There were 18 humeral fractures,5 radioulnar ones,9 femoral ones,and 21 tibial ones.On average,each patient had undergone 1.2 previous operations (from 1 to 2 times),including plating in 29 cases,intramedullary nailing in 15,and external fixation in 9.In the 15 patients whose intramedullary nails were reserved because of slightly loosening only under axial stress,lateral force and rotation,the nonunion was treated with bone autograft + unicortical or full-thickness locking plate.In the other 38 patients whose original implants were removed,the nonunion was treated by re-reaming of the medullary cavity + bone autograft + locking plate internal fixation.Regular radiological follow-ups were conducted for the patients.The functional recovery of upper limbs was evaluated by the Neer system while that of the lower limbs by the Johner-Wruhs system.Results One patient was lost to the follow-up.The rest 52 were followed up for 4 to 15 months (average,12.7 months).Nonunion was found in 2 patients whose internal fixation was broken and loosened.Bone healing was achieved in 50 cases after 4 to 6 months(average,5 months).Removal of implants was conducted in 36 patients at 12 to 24 months after operation (average,13.3 months).At the final follow-ups,the Neer score of upper limb was excellent in 15 cases,good in 5,fair in one and poor in one,giving an excellent to good rate of 90.9％;the Johner-Wruhs score of lower limb was excellent in 24 cases,good in 3,fair in 2 and poor in one,giving an excellent to good rate of 90.0％.Conclusion Locking plate can have good curative efficacy in the treatment of postoperative nonunion of long bone fracture.
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BACKGROUND:The in vivo degradation process of chitosan/hydroxyapatite composite porous scaffolds is not very clear. Research on the effects of rat osteoblasts and degradation products is less. OBJECTIVE:To analyze the biocompatiblity of rat osteoblasts with degradation products of chitosan/hydroxyapatite composite porous scaffolds. METHODS:The second generation of cultured rat osteoblasts were respectively cultured in the extract of degradation products of chitosan/hydroxyapatite composite scaffolds (experimental group) and Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum (control group). At 2, 4, 6, 8, 10 days of culture, cel counting was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Alkaline phosphatase activity was measured by the recommended method of determination of the Federation, and total protein was determined by BCA method. RESULTS AND CONCLUSION:The proliferation speed, alkaline phosphatase activity, total cel ular protein synthesis and ratio of alkaline phosphatase to total protein in rat osteoblasts cultured in the experimental group were significantly higher than those in the control group (P<0.05). This experiment showed that the degradation products of chitosan/hydroxyapatite composite porous scaffolds cannot only promote rat osteoblast adhesion, growth and proliferation, but also enhance its ossification function, with good biocompatibility.
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BACKGROUND:Nowadays, the internal fixation materials for the clinical treatment of unstable femoral intertrochanteric fractures are broadly divided into two categories:intramedul ary fixation system and extramedul ary fixation system. However, the effects of the treatments usual y lack of macro evaluation. OBJECTIVE:To compare the effect of intramedul ary fixation system and extramedul ary internal fixation system in the treatment of unstable femoral intertrochanteric fractures. METHODS:217 patients with unstable femoral intertrochanteric fracture (Evans typing: Ⅲ type to Ⅴ type and R type) were treated by using intramedul ary fixation system and extramedul ary internal fixation system:18 cases were treated with dynamic condylar screw, 67 cases were treated with dynamic hip screw, 43 cases were treated with reconstruction nail, 72 cases were treated with proximal femoral anti-rotation blade nail, and 17 cases were treated with new generation of antegrade interlocking intramedul ary nail. The operative time, length of incision, blood loss, time in bed, fracture healing time, complications and Harris hip score were analyzed and compared. RESULTS AND CONCLUSION:In the aspects of operative time, blood loss, time in bed, complications and Harris hip score, the intramedul ary fixation system was superior to extramedul ary internal fixation system. But there were significant differences in the aspects of length of incision and fracture healing time between two methods. The results showed that the treatment of unstable femoral intertrochanteric fractures by using intramedul ary fixation system was better than the extramedul ary internal fixation system, as the intramedul ary fixation system could effectively shorten the operative time and reduce blood loss. This stable and reliable fixation enables patients to take early postoperative functional exercise, and could significantly shorten the time in bed and reduce the incidence of complications and it is conducive to postoperative hip function recovery. Therefore, intramedul ary fixation system is an ideal method for the treatment of unstable femoral intertrochanteric fractures.
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BACKGROUND:As an oral anticoagulant drug, many experiments have proved that rivaroxaban can prevent the deep venous thromboembolism after the hip arthroplasty. The foreign literatures indicate that it can significantly reduce the incidence of deep venous thromboembolism after extending the treatment course to 35 days. But there is no significant conclusion at home, and the safety of drugs after extentding the course has not been confirmed. OBJECTIVE:To analyze the efficacy and safety of rivaroxaban versus low-molecular-weight heparin for the prevention of deep venous thrombosis after the hip arthroplasty. METHODS:106 patients with primary unilateral hip arthroplasty in the Second Clinical Medical College of Shanxi Medical University between March 2011 and September 2012 were selected. The patients were randomly divided into rivaroxaban group and low-molecular-weight heparin group. The patients in two groups were given drugs at 6 hours after replacement, the patients in the rivaroxaban group were given rivaroxaban 10 mg/d with the course of 5 weeks;the patients in the low-molecular-weight heparin group were given low-molecular-weight heparin 4 100 U/d with the course of 2 weeks. RESULTS AND CONCLUSION:The review and fol ow-up results showed there was no deep vein thrombosis or symptoms of deep vein thrombosis in patients of the rivaroxaban group after replacement, while seven cases (13%) of deep vein thrombosis were observed in the low-molecular-weight heparin group, and there was significant difference in the incidence rate of deep vein thrombosis between two groups (P0.05). The results indicate that ful course of rivaroxaban has clear effect and reliable security in the prevention of deep venous thrombosis after hip arthroplasty.
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ObjectiveTo evaluate the effects of double plates in the treatment of humeral intercondylar fractures.MethodsFrom January 2004 to March 2011,31 patients with humeral intercondylarfractures were reviewed.According to AO/Müller classification,there were three patients with type C1 fractures,16 with type C2 and 12 with type C3.Trans-olecranon osteotomy or trans-triceps-side approach and internal fixation by standard method of AO double plates were performed in all patients.ResultsAll 31patients were followed up for 12 -18 months,which showed fracture healing 6-12 months after surgery.According to the Cassebaum scoring system,the effects were evaluated as excellent in seven patients,good in 19,fair in four and poor in one,with excellence rate of 87％. ConclusionDouble plates is proved to be a good choice for humeral intercondylar fractures,since it can obtain satisfactory outcome with solid fixation,stable mechanical properties,few complications and facilitation for early rehabilitation.