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Objective:To investigate the clinical treatment of large segmental humeral defects with unilateral external fixation and bone transport.Methods:A retrospective study was conducted of the 9 patients who had been treated at Department of Orthopedics, Shenzhen People's Hospital for large segmental humeral defects from September 2017 to June 2019. They were 5 males and 4 females with an average age of 29 years (from 21 to 41 years). Their defects were caused by trauma in 2 cases, by chronic osteomyelitis in 6 cases and by bone tumor in one case. The length of bone defect ranged from 4.2 to 9.0 cm, with an average of 5.9 cm. A unilateral external fixator was placed in operation, and adjusted regularly 7 to 10 days after operation for bone transport and bone lengthening to restore the length of humerus gradually. The external fixation bracket was removed after 3 to 4 layers of cortex were observed on X-ray films. Recorded were length and rate of humeral lengthening, fracture healing time, time for carrying external fixator and complications; the Disabilities of the Arm, Shoulder and Hand (DASH) scores were compared between preoperation and 15 months postoperation.Results:All the patients were followed up for 15 to 36 months (mean, 19 months). The length of lengthening averaged 5.9 cm (from 4.2 to 9.0 cm) with an average lengthening rate of 26%, the healing index 31 d/cm, the bone healing time 8.3 months, and the time for carrying external fixator 10.8 months(from 8.0 to 13.5 months). Their average DASH scores improved significantly from 25.0 ± 2.4 preoperation to 12.0 ± 1.8 at 15 months postoperation ( P<0.05). Good correction of large humeral defects was achieved in all but one case who reported temporary radial nerve paralysis. There were no such complications as neurovascular injury. The shoulder and elbow functions were basically normal after operation. Conclusions:In the treatment of large segmental humeral defects, unilateral external fixation plus bone transport can quickly repair the defects and recover the upper limb function of the patients.
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BACKGROUND: Osteoarthritis is a common cause of pain and disability in the elderly. The underlying cause is the combination of inappropriate mechanical stress, inflammatory mediators and biochemical factors. Curcumin has been shown to have anti-oxidant, anti-inflammatory, anti-bacterial, anti-tumor, neuroprotective, and cardioprotective effects, radiation protection and therapeutic effects on osteoarthritis. Similarly, magnesium sulfate can relieve joint pain and inhibit joint destruction. OBJECTIVE: To investigate the mechanism by which the combined use of curcumin and magnesium sulfate exerts synergistic effect to promote inflammatory chondrocyte reverse differentiation.METHODS: Primary cultured inflammatory chondrocytes were subjected to monolayer culture in vivo. Cell proliferation assay (MTS) was used to detect the proliferation of inflammatory chondrocytes under in vitro monolayer culture conditions. The experimental cells were divided into four groups and underwent 3D induced reverse differentiation culture for 18 days: single culture of chondrocytes (chondrocyte group) , inflammatory chondrocyte cultured with curcumin (curcumin group) , inflammatory chondrocytes cultured with magnesium sulfate (magnesium sulfate group) , and inflammatory chondrocytes cultured with curcumin and magnesium sulfate (combination group). RESULTS AND CONCLUSION: MTS proliferation experiments showed that inflammatory chondrocytes at passage 3 had a higher rate of early proliferation and a lower degree of differentiation. Quantitative PCR results showed that the mRNA levels of type II collagen, proteoglycan and SOX9 were significantly higher in the combination group than in the curcumin group or magnesium sulfate group (P < 0.01). The size of the gross specimens and the positive area of chondrocyte reverse differentiation for alcian blue staining and safranin O staining in the combination group were significantly larger than those in the other three groups (P < 0.01). TUNEL staining results indicated that the positive area of apoptosis-specific staining in the combination group and magnesium sulfate group was significantly smaller than that in the other two groups (P < 0.01). Therefore, the combined use of curcumin and magnesium sulfate has the synergistic effect to promote the reverse differentiation of inflammatory chondrocytes.
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BACKGROUND: Osteoarthritis is a common cause of pain and disability in the elderly. The underlying cause is the combination of inappropriate mechanical stress, inflammatory mediators and biochemical factors. Curcumin has been shown to have anti-oxidant, anti-inflammatory, anti-bacterial, anti-tumor, neuroprotective, and cardioprotective effects, radiation protection and therapeutic effects on osteoarthritis. Similarly, magnesium sulfate can relieve joint pain and inhibit joint destruction. OBJECTIVE: To investigate the mechanism by which the combined use of curcumin and magnesium sulfate exerts synergistic effect to promote inflammatory chondrocyte reverse differentiation.METHODS: Primary cultured inflammatory chondrocytes were subjected to monolayer culture in vivo. Cell proliferation assay (MTS) was used to detect the proliferation of inflammatory chondrocytes under in vitro monolayer culture conditions. The experimental cells were divided into four groups and underwent 3D induced reverse differentiation culture for 18 days: single culture of chondrocytes (chondrocyte group), inflammatory chondrocyte cultured with curcumin (curcumin group), inflammatory chondrocytes cultured with magnesium sulfate (magnesium sulfate group), and inflammatory chondrocytes cultured with curcumin and magnesium sulfate (combination group). RESULTS AND CONCLUSION: MTS proliferation experiments showed that inflammatory chondrocytes at passage 3 had a higher rate of early proliferation and a lower degree of differentiation. Quantitative PCR results showed that the mRNA levels of type II collagen, proteoglycan and SOX9 were significantly higher in the combination group than in the curcumin group or magnesium sulfate group (P < 0.01). The size of the gross specimens and the positive area of chondrocyte reverse differentiation for alcian blue staining and safranin O staining in the combination group were significantly larger than those in the other three groups (P < 0.01). TUNEL staining results indicated that the positive area of apoptosis-specific staining in the combination group and magnesium sulfate group was significantly smaller than that in the other two groups (P < 0.01). Therefore, the combined use of curcumin and magnesium sulfate has the synergistic effect to promote the reverse differentiation of inflammatory chondrocytes.
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Objective To investigate the protective effect of thyroid hormone on spinal cord injury neurons and its molecular mechanism.Methods A DMEM culture medium with a volume fraction of 10％ fetal bovine serum was cultured with the dorsal ridge neurons of RN-dsc rats.The neurons were inoculated in the culture plate after the digestion of trypsin and treated differently as follows:(1) control group:DMEM treatment with no drugs or serum;(2) H2O2 group:serum-free DMEMtreatment containing 100 μmol/L H2O2;(3) H2O2 + 10-6 mol/L triiodothyronine (T3) group:serumfree DMEM treatment containing 100 μnol/L H2O2 and 10-6mol/L T3;(4) H2O2 + 10-5 mol/L T3 group:serum-free DMEM treatment containing 100 μ mol/L H2O2 and 10-5 mol/L T3;(5) negativecontrol group:transfection of negative control mimics with LipofectamineTM2000 reagent;(6) miR-210 group:transfection of miR-210 mimics with LipofectamineTM 2000 reagent.Cell viability,apoptosis number,and expressions of nuclear factor E2 correlation factor 2 (Nrf-2) antioxidant pathway molecules and miR-210 were determined.After transfection of miR-210 mimics and negative control mimics,expressions of Nrf-2 antioxidant pathway molecules were determined.Results The cell proliferation activity and protein expressions of Nrf-2,antioxidant reaction elements (ARE),superoxide dismutase 2 (SOD2),and heme oxygenase (HO-1) in H2O2 group (0.39 ±0.06,0.52 ±0.08,0.31 ±0.08,0.25 ± 0.05,respectively) were significantly lower than those in control group (1.00 ± 0.15,1.00 ± 0.17,1.00 ± 0.13,1.00 ± 0.11,respectively) (P ＜ 0.05),while the apoptosis numbers and the expressions of miR-210 were significantly higher than those in control group (P ＜ 0.05).The cell proliferation activity and protein expressions of Nff-2,ARE,SOD2,HO-1 in H2O2 + 10-6mol/L T3 group and H2O2 +10-5 mol/L T3 group were significantly higher than those in the H2O2 group (P ＜ 0.05),while apoptosis numbers and expressions of miR-210 were significantly lower than those in H2O2 group (P ＜ 0.05).The cell proliferation activity and protein expressions of Nrf-2,ARE,SOD2,HO-1 in H2O2 + 10-5mol/L T3 group (0.88 ±0.14,0.84 ±0.12,0.72 ±0.09,0.69 ±0.09) were significantly higher than those in H2O2 + 10-6mol/L T3 group (0.73 ±0.09,0.71 ±0.08,0.58 ±0.09,0.52 ±0.08) (P＜0.05),while apoptosis numbers and expressions of miR-210 were significantly lower than those in H2O2 + 10-6mol/L T3 group.The protein expressions of Nrf-2,ARE,SOD2,and HO-1 in miR-210 group (0.37 ±0.06,0.24 ±0.05,0.45 ± 0.08,0.49 ± 0.07,respectively) were significantly lower than those in negative control group (1.00±0.13,1.00±0.19,1.00±0.15,1.00±0.14,respectively) (P＜0.05).Conclusion Thyroid hormone can inhibit the expression of Nrf-2 in oxidative stress injury process of neurons by inhibiting the expression of miR-210,and hence reduce the oxidative stress injury of spinal cord neurons.
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Objective To explore the eftect of SS31 peptide on autophagy after spinal cord injury (SCI) and possible mechanism.Methods Allen nethod was used to construct the spinal cord injury model in rats.Sprague-Dawley rats were randomly divided into sham surgery group (sham group),SCI group and SS31 peptide group,with 30 rats in each group.The sham group only received laminectomy.The rats in SCI group were sustained SCI and were given no intervention.The rats in SS31 group received SS31 peptide injection after SCI.Scores of Basso Beattie Bresnahan (BBB) motor functions were assessed at 6 h,1,3,7 and 14 d after the injury.The changes in related proteins of Beclin-1 and LC3-Ⅱ were also detected.Results Scores of BBB scale at 6 h and at days 1,3,7 and 14 after injury in SCI group (0,1.7 ±0.4,3.5 ±0.6,6.1 ±0.7,10.1 ±0.6) and SS31 peptide group (0,2.5 ±0.7,4.1 ±0.7,9.3 ±0.6,13.4 ±0.6) were lower than that in sham group (21 at all time points) (P ＜0.05).Scores of BBB scale at days 7 and 14 after injury in SS31 peptide group (9.3 ±0.6,13.4 ±0.6) was higher than that in SCI group (6.1 ± 0.7,10.1 ± 0.6) (P ＜ 0.05).There was no significant difference upon scores of BBB scale of SS31 peptide group at 6 h and at days 1 and 3 after injury (0,2.5 ±0.7,4.1 ± 0.7),compared with SCI group (0,1.7 ± 0.4,3.5 ± 0.6) (P ＞ 0.05).Compared with sham group,the expression of Beclin-1 in SCI group and SS31 peptide group was increased,reached a peak at day 3 (1.478 ± 0.030,1.841 ± 0.051),remained high level at day 7 (1.302 ± 0.049,1.551 ± 0.032) and showed high expression at day 14 (1.252 ±0.048,1.471 ± 0.062) (P ＜ 0.05).Compared with sham group,the expression of LC3-Ⅱ in SCI group and SS31 peptide group also increased,reached a peak at day 3 (0.348 ± 0.028,0.655 ± 0.052),remained high level at day 7 (0.301 ± 0.053,0.432 ± 0.052) and also showed high expression at day 14 (0.268 ± 0.049,0.371 ± 0.052) (P ＜ 0.05).The expressions of Beclin-1 and LC3-Ⅱ in SS31 peptide group at day 3 after injury were 1.841 ± 0.051 and 0.455 ± 0.052,higher than that in SCI group (1.478 ± 0.030,0.348 ± 0.028) (P ＜ 0.05).In SS31 peptide group at 6 h and days 1,7 and 14 after injury,the expressions of Beclin-1 (0.582 ± 0.028,0.723 ±0.049,1.551 ±0.032,1.471 ±0.062) and LC3-Ⅱ (0.172 ±0.031,0.256 ±0.051,0.432 ± 0.052,0.371 ± 0.052) had no significant difference in comparison with corresponding expressions of Beclin-1 (0.584 ±0.021,0.642 ±0.051,1.302 ±0.049,1.252 ±0.048) and LC3-Ⅱ (0.156 ± 0.019,0.184±0.050,0.301 ±0.053,0.268 ±0.049) in SCI group (P＞0.05).Conclusion SS31 peptide can improve motor function and enhance the autophagy of nerve cells after SCI in rats,which may be one of the mechanisms for SS31 peptide treating spinal cord injury.
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BACKGROUND: Multipotent differentiation ability enables mesenchymal stem cells from autologous bone marrow to differentiate into osteoblasts and chondroblasts, thereby promoting the formation of bones and cartilage. However, the osteogenic ability differs from each other, and whose osteogenic ability is the best still needs to be studied further. OBJECTIVE: To compare the osteogenic ability of fascia- and muscle-derived stem cells in rats. METHODS: Fascia- and muscle-derived cells were isolated from 20 rats, followed by flow cytometry sorting, and were then cultured. FDC-LacZ cells were transfected with retro-BMP4 virus twice. Afterwards, the transfection efficiency of fascia-derived cells was detected through LacZ and alkaline phosphatase staining. RESULTS AND CONCLUSION: Compared with fascia-derived cells, muscle-derived cells showed stronger chondrogenic ability and produced more calcium deposition. These findings indicate that the osteogenic ability of muscle-derived cells is superior to that of fascia-derived cells in rats.
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BACKGROUND:Mesenchymal stem cells from human skeletal muscle exhibit multi-directional differentiation potential under the influence of osteogenic proteins such as bone morphogenetic protein 4 (BMP4). But the differentiation of a specific cell subpopulation is not yet clear.OBJECTIVE:To characterize the multi-directional differentiation potential of mesenchymal stem cells from human skeletal muscle based on the expression of different surface markers.METHODS:Four different subpopulations were isolated from the human skeletal muscle by fluorescence-activated cell sorting based on their expression of the myogenic-specific marker CD56 and the endothelial-specific markers CD34 and CD144, including CD56+, CD56+CD34+CD144+, CD34+CD144+, and unsorted groups. Osteogenic differentiation of the four groups of the cells was displayed by Von Kossa staining after the treatment with BMP4 alone or BMP4 plus transforming growth factor β3. Chondrogenic differentiation of these cells was displayed by Alcian blue staining. Bone metabolism was assessed by alkaline phosphatase staining.RESULTS AND CONCLUSION:No significant difference in the bone metabolism was found among four groups after the treatment with BMP4 (P > 0.05). Osteogenic and chondrogenic potentials of the four cell subpopulations were significantly different. Under the same osteogenic induction, the CD56+ cells exhibited strongest potential for osteogenic differentiation; and under the same chondrogenic induction, the CD56+CD34+CD144+ cells exhibited better potential for chondrogenic differentiation than the CD56+ cells. These findings indicate that the osteogenic and chondrogenic potentials are intimately associated with the type of mesenchymal stem cells from human skeletal muscle:the CD56+ cells are closely related to the osteogenic potential, while the CD56+CD34+CD144+ cells have stronger chondrogenic potential.
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Knee Osteoarthritis is one of the most common degenerative diseases.Although the morbidity of knee osteoarthritis increases dramatically,there is no effective medication that can repair the damaged articular cartilage and restore the joint function.Recently,with the technical development of cell transplantation and regulation,more attentions have been paid in this area.The techniques included osteochondral autograft transplantation,mosaicplasty,chondrocytes based matrix-induced autologous chondrocyte implantation,and mesenchyma stem cells based cell therapy.The study of cell differentiation is also more in-depth.It has been confirmed that all of these methods can alleviate the symptoms and improve the function of knee osteoarthritis to some extent.Stem cell therapy seems have greater advantage than other intervention.Massive chondrocytes can be gained through induction and culture of mesenchyma stem cells in vitro,and regenerated cartilage could be found on the surface after delivering stem cells into knee joint through a certain way.These technologies are hopeful to prevent from knee osteoarthritis or even reverse the progress of it at early stage.However,there are a variety of problems about in vitro cell transplantation,such as the unknowing regulation of mesenchyma stem cells,the difficulty of keeping the characteristic stability of new born chondrocytes,the integrity of new cartilage tissue and the way chondrocytes delivered.Cell therapy has been applied both in basic research and in clinic trials.The present review is focused in the current progress of cell therapy in knee osteoarthritis and discuss the challenges and troubles.Furthermore,we also summarize a series of ongoing clinical trials,and try to find out the exact clinical effects of cartilage repair by using cell therapy.
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Objective To produce bioactive human bone morphogenetic protein-4 by Escherichia coli genetic engineering and investigate the effect of the product, recombinant BMP-4, on the bone marrow stem cells. Methods cDNA of human morphogenetic protein-4 mature peptide was obtained by RT PCR from tissue of human placenta. The gene was constructed in the pET-22b(+) expression vector and expressed in Escherichia coli BL-21 after transformation and induction by IPTG. The harvested protein was proved to be bioactive by inducing ectopic bone information in mouse thigh. The protein was applied to induce the cultured bone marrow stem cell. Shape change of the cell, ability of ALP (alkaline phosphatase) and concentration of OC (osteocalcin) were investigated. Results SDS-PAGE revealed a new protein band that located in position of 14?103 after 4 hours induction, the new protein made 15% of total bacteria protein, the rhBMP-4 could induce the cultured bone marrow stem cell of New Zealand rabbit to differentiate into osteoblasts and form calcified node. The ability of ALP and concentration of OC of tested group increased significantly than that of the control group. Conclusion The bioactive rhBMP-4 can be produced by Escherichia coli genic engineering, this protein can induce bone marrow stem cells to differentiate into osteoblast cells.