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Bone defects caused by different causes such as trauma, severe bone infection and other factors are common in clinic and difficult to treat. Usually, bone substitutes are required for repair. Current bone grafting materials used clinically include autologous bones, allogeneic bones, xenografts, and synthetic materials, etc. Other than autologous bones, the major hurdles of rest bone grafts have various degrees of poor biological activity and lack of active ingredients to provide osteogenic impetus. Bone marrow contains various components such as stem cells and bioactive factors, which are contributive to osteogenesis. In response, the technique of bone marrow enrichment, based on the efficient utilization of components within bone marrow, has been risen, aiming to extract osteogenic cells and factors from bone marrow of patients and incorporate them into 3D scaffolds for fabricating bone grafts with high osteoinductivity. However, the scientific guidance and application specification are lacked with regard to the clinical scope, approach, safety and effectiveness. In this context, under the organization of Chinese Orthopedic Association, the Expert consensus for the clinical application of autologous bone marrow enrichment technique for bone repair ( version 2023) is formulated based on the evidence-based medicine. The consensus covers the topics of the characteristics, range of application, safety and application notes of the technique of autologous bone marrow enrichment and proposes corresponding recommendations, hoping to provide better guidance for clinical practice of the technique.
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Objective:To compare the clinical effects of discectomy combined with transpedicular dynamic stabilization and transforaminal lumbar interbody fusion (TLIF) in treating single-level lumbar disc herniation.Methods:From November 2012 to November 2015, a total of 96 patients with single-level lumbar disc herniation (disc height decreased more than 1/3, the width of the basilar part of the herniated disc >6 mm, massive disc herniation or Modic type I endplate changes) treated by discectomy combined with Dynesys dynamic stabilization (Dynesys group, n=48) or TLIF (fusion group, n=48) were enrolled. Clinical assessments included operation duration, intraoperative blood loss, MacNab score, visual analogue scale (VAS), Oswestry disability index (ODI) and rate of complications. Radiographs were evaluated for lumbar mobility, intervertebral height, etc. Results:A total of 86 patients were included in the final analysis (44 in Dynesys group and 42 in fusion group) and were evaluated after 5 years follow-up. The operation duration of Dynesys group (159.61±37.29 min) was less than that of the fusion group (177.42±39.90 min) significantly ( t=2.140, P=0.035). Intraoperative blood loss in Dynesys group (151.78±50.88 ml) was less than that in fusion group (197.74±76.55 ml) with significant difference ( t=3.293, P=0.001). At 5 years follow-up, there were 2 cases with screw loosening and 5 cases with adjacent segmental degeneration in Dynesys group without symptom. In fusion group, there were 12 cases with adjacent segmental degeneration and two of them with symptom. There were significant differences in the incidence of adjacent segment degeneration between the two groups ( χ2=4.012, P=0.045). According to the MacNab criteria, excellent or good cases accounted for 95% in Dynesys group and 93% in fusion group without significant differences ( Z=0.425, P=0.671). VAS back, VAS leg and ODI scores were improved significantly in both groups after 2 years and 5 years ( P<0.05). However, there were no significant differences between the two groups ( P<0.05). The activity of the surgical segment was 4.59°±0.48° in Dynesys group and 1.00°±0.42° in fusion group at 5 years after surgery. The height of intervertebral space in Dynesys group decreased from 11.19±2.07 mm before surgery to 9.98±2.02 mm at 2 years after surgery and to 9.86±1.64 mm at 5 years after surgery ( F=6.462, P=0.002). However, there was no statistically significant difference between the 2 and 5 years follow-up ( q=0.415, P>0.05). At 5 years after surgery, the activity of the first proximal segment in the two groups was 9.74°±3.29° and 11.69°±3.89°, respectively ( t=2.514, P=0.014). Conclusion:Both discectomy combined with dynamic stabilization and TLIF can achieve satisfied clinical effects in treating single-level lumbar disc herniation. Dynamic stabilization preserves the intervertebral activity of surgical segments and results in a lower incidence of adjacent segment degeneration compared with that in fusion surgery. Furthermore, discectomy combined with dynamic stabilization is a less invasive intervention with shorter operation duration and less blood loss compared with TLIF.
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Objective:]To investigate the role of Arpin protein in bone repair by mediating migration of host bone marrow mesenchymal stem cells (BMSCs) to the bone defect area after transplantation of tissue engineering bone (TEB).Methods:Immunofluorescence was used to observe the expression and relative localization of Arpin and Arp2/3 proteins in BMSCs. Lentiviruses that ware designed to interfere with Arpin expression were constructed to transfect BMSCs for knockdown Arpin expression. Knockdown efficiency was verified by real-time quantitative reverse transcription PCR ( qRT-PCR) and Western blot. According to different levels of Arpin protein expression, experiments were divided into empty vector control group and an Arpin expression inhibition group in vitro and in vivo. In vitro experiments: the cell migration model was established with a migration chamber, then the cells from both groups were seeded on the up chamber, and the number of migrated cells were detected by fluorescence microscopy. Cells from both groups were seeded on six-well plates. Model of wound healing experiment was established and wound healing ratio was examined by microscopy. In vivo experiments: 8-week-old C57BL/6 mice were selected and assigned to empty vector control group and Arpin expression inhibition group according to the random number table, with 6 rats per group. Diaphysis of 2 mm and periosteum in the middle femur were excised to make a large segment of bone defects. Then, TEB was transplanted into the defect area and fixed.Green fluorescein-labeled BMSCs (1 million cells per mouse) from empty vector control group and Arpin expression inhibition group were injected through the tail vein. Number of BMSCs homing to the bone defect area was detected by immunofluorescence staining at day 2 and 7 after operation. At 4 weeks after operation, the femur was taken for a Micro-CT scan to analyze bone mass density(BMD), bone volume density (BV/TV), trabecular spacing (Tb.Sp) and trabecular thickness (Tb.Th). Then, the specimens were stained with pathological HE and MASSON staining to observe the quality of bone formation. Results:Mouse BMSCs expressed Arpin protein, which was located at the cell edge relative to Arp2/3. After transfection of lentivirus, BMSCs expressed green fluorescent protein, and the expression of Arpin gene and protein in Arpin expression inhibition group were decreased compared to empty vector control group ( P<0. 01). BMSCs migration was enhanced in Arpin expression inhibition group compared to empty vector control group [(76.6±6.6) vs. (105.7±6.5)] ( P<0. 01). Wound healing was accelerated in Arpin expression inhibition group compared to empty vector control group [(43.8±0.19)% vs. (62.6±3.2)%]( P<0.01). At day 2 after operation, immunofluorescence results showed no significant difference in cell migration between the two groups and almost no labeled cells migrated. At day 7 after operation, more cells migrated to the transplanted area in Arpin expression inhibition group compared to empty vector control group [(5.7±1.5) vs. (11.3±1.5)] ( P<0.01). At 4 weeks after operation, Micro-CT results showed that Arpin expression inhibition group had better bone formation quality than empty vector control group [BMD: (172.7±6.0)mg/cm 3vs. (140.0±6.0)mg/cm 3, BV/TV: (28.8±1.3)% vs. (23.4±0.9)%, Tb.Sp: (0.33±0.01)μm vs. (0.28±0.01)μm, Tb.Th: (0.11±0.01)μm vs.(0.15±0. 01)μm]( P<0.05). Pathological staining showed there were more new bone tissue in Arpin expression inhibition group ( P<0.01). Conclusion:Silencing Arpin protein expression promotes BMSCs to migrate to the bone defect area and improves bone repair effect.
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Objective To investigate the mechanism of implanted tissue-engineered bone (TEB)recruiting endogenous mesenchymal stem cells (BMSCs) towards bone regeneration after traumatic bone defect.Methods In vivo experiments:2 mm of diaphysis and periosteum were removed from the middle of the femoral shaft in 8 week old FVB/N mice to form a large segment of bone defect.Demineralized bone matrix (DBM) and TEB were implanted into the defect area and fixated.All mice were randomly divided into DBM group (n =18) and TEB group (n =18).The results were observed 24 hours after implantation:(1) flow cytometry was used to evaluate the number of mobilized host BMSCs into the blood;(2) non-invasive bioluminescent imaging was used to observe the ability of two groups in recruiting mouse bone marrow derived mesenchymal stem cells (mBMSCs) in peripheral blood to the defect area;(3) ELISA was used to evaluate the stromal cell-derived factor 1 (SDF-1) content in peripheral blood of two groups.In vitro experiments:(1) transwell assay was conducted to evaluate the ability of SDF-1 (100 ng/ml) in promoting the migration of human bone marrow derived mesenchymal stem cells (hBMSCs).SDF-1/C-X-C motif chemokine receptor-4 (CXCR4) pathway was blocked by the selective CXCR4 antagonist Plerixafor (AMD3100).The experimental groups were divided into control group,SDF-1 group,and SDF-1 + AMD3100 group.(2) The co-culture system of human umbilical vein endothelial cells (hUVECs) and hBMSCs was established,and cells were stimulated by SDF-1.The experimental groups were divided into hBMSCs group,hBMSCs + hUVECs group,and hBMSCs + hUVECs (AMD3100 pretreatment) group.Transwell assays were used to compare the migration of hBMSCs in each group.ELISA was used to detect the concentration of hepatocyte growth factor (HGF) in the co-culture supernatant.(3) In vitro cultured hUVECs were stimulated by SDF-1 and SDF-1/CXCR4 pathway was antagonized by AMD3100.The experimental groups were divided into control group,SDF-1 group,and SDF-1 + AMD3100 group.Quantitative real-time polymerase chain reaction (qRT PCR) was used to evaluate the expression of HGF in each group.Results In vivo experiments:24 h after transplantation,the number of BMSCs and SDF-1 concentration in the TEB group were significantly highcr than those in the DBM group (P ＜ 0.05).The number of recruited mBMSCs into the circulation in the TEB group was larger than that in the DBM group (P＜ 0.01).In vitro experiments:(1) compared with the control group and the SDF-1 + AMD3100 group,the SDF-1 group significantly enhanced the migration ability of hBMSCs in Transwell migration experiments (P ＜ 0.01);(2) compared with the hBMSCs group and the hBMSCs + hUVECs (AMD3100 pretreatment) group,the number of migrated cells and HGF concentration in the hBMSCs + hUVEC group significantly increased (P ＜ 0.01),but there were no significant differences between the hBMSCs group and the hBMSCs + hUVECs (AMD3100 Pretreatment) group (P ＞0.05);(3) qRT-PCR showed that the expression of HGF was significantly increased in the SDF-1 group compared with the control group (P ＜ 0.05).After antagonizing SDF-1/CXCR4,HGF expression in the SDF-1 + AMD3100 group was significantly lower than that in the SDF-1 group.Conclusions TEB transplantation in traumatic bone defect can significantly increase the concentration of chemokine SDF-1 in vivo and effectively promote the mobilization of endogenous MSCs and recruitment of circulating MSCs.SDF-1 not only directly promotes the migration of hBMSCs through SDF-1/CXCR4 pathway,but also up-regulates the expression and secretion of HGF in vascular cells to further amplify the chemotactic effect of SDF-1 on hBMSCs.
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Objective To analyse the phenotypes of the drug?resistant tuberculosis, and investigate the outcomes of the individualize surgery and chemotherapy for these patients. Methods From January 2009 to June 2012, we retrospectively ana?lyzed 49 patients with drug?resistant tuberculosis spondylitis admitted in Southwest Hospital. 33 were initial cases and 16 were re?curring cases. All the 49 patients received individualized open operation or CT?guided percutaneous drainage and local chemother?apy depending on the characteristics of the focus. Individualized chemotherapy regimens were tailored for all patients according to the drug?resistant spectrum and all patients were followed up successfully at least 24 months. All the clinical data were collected and analyzed by statistical methods. Results Among the 49 patients, 14 were monoresistance tuberculosis, 11 were polyresis?tance tuberculosis, and 24 cases were multi?drug resistant tuberculosis. Frequence of the drug?restistance from high to low was Iso?niazid, Rifampicin, Streptomycin, Levofloxacin, Dipasic/Rifapentine, Ethambutol, Protionamide, Capreomycin, Paza?aminosalicy?late, and Amikacin. 43 patients received open operation and 6 patients received CT?guided percutaneous drainage and local che?motherapy. Time of the percutaneous drainage was (48±11) days (39-60 days), and all patients received Individualized chemother?apy with an average of (29.5±2.5) months (24-36 months) postoperatively. At the last follow?up, all patients had remarkable pain remission, 44 patients with paraplegia got slight or remarkable recovery and 17 patients with kyphosis got significant correction. Conclusion The main drug?resistant spectrums are Isoniazid、Rifampicin、Streptomycin、Levofloxacin. The individualized sur?gery combined with individualized chemotherapy made according to the drug?resistance is a feasible treatment for the drug?resis?tant tuberculosis especially the multi?drug resistant tuberculosis.
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Objective To study the efficacy and safety of four surgical techniques of tuberculosis of lumbosacral junction retrospectively. Methods Between Jul 2001 and Jan 2013, 79 patients with lumbosacral spinal tuberculosis underwent surgery. Antituberculous chemotherapy and nutrition support prior to surgery were used for at least two weeks. 45 patients underwent single stage radical debridement, fusion and anterior instrumentation (A group). 18 patients underwent combined anterior and posterior spinal surgery (AP group), 10 patients underwent transpedicular drainage, posterior instrumentation, and fusion (P group), and 6 patients underwent anterior radical debridement (D group). All the patients were treated by antituberculous chemotherapy for 18 months and followed regularly. The operation duration, blood loss, clinical status, ESR, VAS, ODI, roentgenogram and 3D?CT were concerned to estimate the progress of tuberculosis. Radiographs were analyzed before surgery, immediately after surgery, and at the final follow?up examination to assess the result of anterior fusion and maintenance of correction. Results There was no inju?ry of blood vessel, ureter or cauda equina during surgery. The mean follow?up period was 23 months (range 18-42 months). No obvious loss of deformity correction was observed. There was no recurrence, no tuberculous peritonitis, and no incidence of im?potence or retrograde ejaculation in any of these patients. The average operating duration(min) were 144.31 ± 23.18, 444.72 ± 141.63, 351.50 ± 85.25, 90.00 ± 29.66, respectively; The average blood loss(ml)were 266.67 ± 104.45, 988.99 ± 488.26, 890.00 ± 306.23, 200.00±104.88, respectively; The average Pre?op VAS were 4.71±1.79, 5.22±1.48, 3.30±1.64, 2.50±1.52, respectively;The average last follow?up VAS were 0.89±0.68, 0.90±0.74, 1.00±0.63, respectively; The average Pre?op ODI(%)were 29.64± 7.85, 32.17±7.59, 28.20±4.26, 20.67±4.63, respectively; The average last follow?up ODI(%)were 5.09±3.59, 4.78±3.78, 4.80± 3.39, 4.00 ± 1.18, respectively; The average Pre?op lumbosacral angle(°)were 20.61 ± 4.92, 23.78 ± 5.84, 25.10 ± 4.28, 21.67 ± 4.27, respectively; The average Post?op lumbosacral angle were 27.17±3.66, 30.56±5.31, 32.10±4.01, 24.83±2.32, respectively;The average last follow?up lumbosacral angle were 23.89 ± 3.12, 27.00 ± 5.46, 29.00 ± 4.85, 23.33 ± 2.50, respectively. Conclu?sion Single stage anterior interbody fusion with anterior instrumentation worked effectively to stabilize lumbosacral junction (less invasive, short surgical duration, no injury of posterior column). Anterior interbody fusion combined with posterior instrumentation was recommended for patients with extensive bone defect and low iliocava junction.
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Objective To establish the anti-infective tissue engineered bones (TEBs) and evaluate the anti-infective and osteogenic effects of the infection-prevention TEBs on femoral large bone defects in goats.Methods Based on the controlledrelease antibiotic system fibrin gel-coated vancomycin alginate beads (FG-Vanco-AB),the infection-prevention TEBs were established and evaluated.They were transplanted into the critical-size defects in the right femurs of goats.TEBs without the controlled-release antibiotic system were used as controls and transplanted into the left femoral defects.The breakpoint sensitivity of vancomycin (5 mg/mL) for S.aureus was used as a standard concentration.Postoperatively,the vancomycin concentrations in the lesion site,in the adjacent site and in the circulation,as well as the anti-infective effects of the infection-prevention TEBs were evaluated by High-performance liquid chromatography (HPLC).Bone hcaling was assessed by histology,CT and ECT.The results were used to evaluate the osteogenic effect of the infection-prevention TEBs.Results Results from ESM,CLSM and in vivo tracing showed that the in vitro and in vivo survival conditions of seeded cells were analogous to those of TEBs.The effective concentration (over the bactericidal concentration) of vancomycin in bilateral defects and in blood lasted for 28 days,2 days and 7 days,respectively.The concentration of vancomycin in the femur decreased gradually from the grafted site to both ends.At 28 and 56 days postoperatively,the ECT results showed no significant difference between the right and left femurs.CT and histology demonstrated that at 14,28 and 112 days after surgery,bone defects in the bilateral femurs were repaired synchronously,and were completely covered by new bone tissue after 112 days.Conclusion The anti-infective TEBs were successfully established.FG-Vanco -AB in the transplanted sites provided the local bone tissues with anti-infective capability whilst not interfered the process of bone reconstnction and wound healing.
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Objective To evaluate the long-term clinical efficacy of iliac bone block allograft in anterior reconstruction of thoracolumbar fractures.Methods A follow-up study was carried out on 93 patients with thoracolumbar fractures treated by anterior decompression and interbody fusion with iliac bone block allograft from February 2004 to March 2007.The radiographic and clinical outcomes were retrospectively assessed.Results Sixty-six patients were followed up for 60-97 months (mean,78 months).The effective interbody fusion rate was 100％,with the fusion time of 6-12 months (mean,10 months).The Cobb' s angle corrected from pre-operative (21.6 ± 8.3)° to post-operative (5.8 ±5.2)°,but when the infusion became effective,the angle showed average loss of 2.4°.All the 66 patients were associated with various degrees of neurological deficiency,but the patients with incomplete nerve dysfunction obtained different degree of recovery.Conclusions Iliac bone block allograft is effective in reconstruction of spinal anterior-middle column stability with the aid of internal fixation instruments.Thereby,it may be a potential alternative to autograft in the treatment of thoracolumbar fractures and can attain early and late stage persistent spinal stability.
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Objective To evaluate the efficacy of internal fixation with or without fusion in the treatment of thoracolumbar burst fractures.Methods Clinical controlled trails related to the application of pedicle screw instrumentation with or without fusion for thoracolumbar fractures before March,2012were obtained by searching PubMed,Science Direct,Medline and CNKI.Quality evaluation was made on the included literature,from which data were extracted to integrate various rescarch results by using RevMan 5.1.The quantitative data were analyzed based on the effect scale of mean difference (MD) and bilateral 95％ confidence interval (CI).The numeration data were analyzed in the use of effect scale of odds ratio (OR) and bilateral 95％ CI.The merging of some data was manually completed.Results After retrieving,eight English and one Chinese papers of the clinical controlled trials,and two related Meta analysis were obtained.With exclusion of one repetitive research,eight papers were involved in the review.Meta analysis demonstrated that fusion and non-fusion fixation had no significant differences in aspects of correction of kyphotic angle,correction and correction loss of vertebral body height,neurological function improvement,complication rate,and length of hospital stay.While compared with the fusion fixation,non-fusion fixation showed a more serious correction loss of kyphotic angle,a fewer blood loss and a shorter operation time.Conclusions Non-fusion fixation shows the similar efficacy with fusion fixation in the treatment of some thoracolumbar burst fractures pertaining to releasing compression,restoring spinal stability and preventing complications,but it can also significantly decrease operation time and blood loss.Furthermore,non-fusion fixation may markedly improve patients' quality of life since it restores motion of the instrumented segment after removal of implant and decreases the risk of adjacent segmental degeneration.
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Objective To observe the osteogenic effect of tissue-engineered bone constructed by poly-L-lysine-demineralized bone matrix (PLL-DBM) enriched bone marrow stem cells in the space of goat transverse process bone fusion model and explore a new tissue-engineered bone construction method. Methods PLL was used to decorate goat DBM to prepare a matrix material (PLL-DBM). The osteo-genic effect of tissue-engineered bone constructed by PLL-DBM enriched bone marrow cells ( Group Ⅰ A) was detected in goat lumbar intertransverse graft bone model; autogenous iliac bone (Group Ⅰ B), DBM enriched bone marrow (Group Ⅱ C) and DBM (Group Ⅱ D) were used as controls. The osteogenesis of the bones in the fused segments of four groups were compared and evaluated by X-ray, three-dimensional CT, CT value testing and biomechanical testing. Results The results of X-ray showed that the fusion ranges in groups ⅠA and ⅠB were basically the same, which were significantly wider than that in Group Ⅱ, with no fusion detected in Group Ⅱ D. The CT value was (696.76±10275) HU in Group Ⅰ A and (766.03±69.24) HU in Group B, which were significantly higher than that in Group Ⅱ C (P <0.05), but there was no statistical difference in CT value between Groups Ⅰ A and Ⅰ B (P > 0.05). The CT val-ue in Group Ⅱ C was significantly higher than in Group ⅡD (P <0.01). There was no statistical differ-ence between Groups Ⅰ A and Ⅰ B in the maximum load and bending strength (P > 0.05). The maxi-mum load and bending strength in Groups Ⅰ A and Ⅰ B were significantly higher than that in Group Ⅱ C (P < 0.05), and the two indices in Group Ⅱ C were significantly higher than that in Group Ⅱ D (P <0.01). Conclusion Tissue-engineered bone constructed by PLL-DBM enriched bone marrow cells is an ideal tissue engineered bone and its osteogenic potential is similar to that of autologous bone.
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Objective To prepare and optimize fibrin-gel-coated vaneomycin alginate beads (FG-Vanco-AB)and investigate their possible use in treatment of osteomyelitis or prevention of infection.Methods Vancomycin alginate beads were produced by dropping vancomycin and alginate mixed liquor into calcium chloride solution.Beads including high vancomycin content were prepared and chosen by optimizing different concentrations of vancomycin solution and alginate solution.These beads were coated with fibrin gel formed by different concentrations of fibrin and the same concentration thrombin.The optimized beads were selected based on available release time,when vancomycin in medium could kill Staphylococcus aureus(ATCC25923). Results Higher content of vancomycin in bead resulted in increase of vancomycin concentration and alginate concentration in mixed liquid.The highest vancomycin content beads were prepared by 16%alginate and 50 mr/ml vancomycin,up to(27.36±0.90)%.The further results showed that vancomycin concentrations from beads coated with fibrin at 75 mg/ml and thrombin at 400 IU/ml could kill Staphylococcus aureus and remained above the breakpoint sensitivity for 19 days.Conclusion The available release time is prolonged,and the possibility of clinical use is conspicuously increased after vancomycin beads are optimized by adjusting the rate of mixed component and fibrin gel coat.