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Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.
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Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.
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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 investigate the treatment effect of marrow mesenchymal stem cells (MSC)/endothelial progenitor cells (EPC) extracellular matrix-based tissue engineering bone (ECM-TEB) in repair of femoral defects in rats.Methods:Bone marrow-derived MSC and EPC were isolated and cultured for functional identification, and planted on the nanocrystalline collagen-based artificial bone particles. After culturing for 14 days, the cells were lyophilized to obtain MSC/EPC ECM-TEB and MSC ECM-TEB. A scanning electron microscope was used to observe the morphology of MSC and EPC on the surface of the scaffold. The protein extracts of MSC ECM-TEBs (control group) and the protein extracts of MSC/EPC ECM-TEBs (experimental group) were added to the EPC culture system for migration test, scratch repair assay, and tube formation detection; and to the MSC culture system for alizarin red staining and alkaline phosphatase? (ALP) staining detection. The cell recruitment, angiogenesis and osteogenic differentiation were observed. A total of 12 SD rats were selected to establish a femoral defect model. According to the random number table, the rats were divided into: (1) sham group: debridement treatment was performed only at the defect; (2) MSC ECM -TEB group: MSC ECM-TEB was implanted at the defect; (3) MSC/EPC ECM-TEB group: MSC/EPC ECM-TEB was implanted at the defect, with 4 rats per group. Two months later, micro-computed tomography (Micro-CT) and Masson's tricolor staining were performed to observe the treatment effect of the bone defect. When the cells were stored at low temperature for three months after lyophilization, the different protein profile between MSC ECM-TEB and MSC/EPC ECM-TEB in vascularization was detected by isotope relative labeling and absolute quantification technology (iTRAQ). The gene ontology/Kyoto Encyclopedia of Gene and Genome Technology (GO/ KEGG) function enrichment was used to analyze the key differences.Results:MSC and EPC grew well and formed a smooth cell layered structure on the surface of the scaffold. The number of cell migration, ratio of scratch repair, and length of the tube in experimental group were respective 121.6±8.3, (61.5±5.9)%, (11.3±0.6)mm, significantly increased compared with control group [85.0±6.7, (39.3±3.6)%, (5.9±0.4)mm] (all P<0.01). Alizarin red staining and ALP staining results showed that the proportion of calcium nodule mineralized area in experimental group increased significantly compared with control group [(38.8±3.3)%∶(49.9±3.0)%, (38.8±2.4)%∶(45.3±3.3)%] (all P<0.05). Base on the Micro-CT and Masson staining, bone defect healing was good in MSC/EPC ECM-TEB group, only a small amount of new bone was formed in MSC ECM-TEB group, and there was almost no new bone regenerated in sham group. Significant differences were found in bone volume/total volume, trabecular number and trabecular thickness among groups (all P<0.05), which were in line with Micro-CT and Masson staining results. The protein profile analysis showed that 83 angiogenesis-related factors in MSC/EPC ECM-TEB group were significantly up-regulated compared with MSC ECM-TEB group (fold change>2, P<0.05). GO/KEGG function enrichment analysis showed that MSC/EPC ECM-TEB group had projecting ascendancy in "vascular development" and in "vascular smooth muscle contraction pathway" compared with MSC ECM-TEB group (both P<0.01). Conclusion:MSC/EPC ECM-TEB has advantages in cell recruitment, angiogenesis, and new bone formation compared with MSC ECM-TEB, and is a better construction strategy for repair of traumatic bone defect.
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TSCI have dyskinesia and sensory disturbance that can cause various life-threaten complications. The patients with traumatic spinal cord injury (TSCI), seriously affecting the quality of life of patients. Based on the epidemiology of TSCI and domestic and foreign literatures as well as expert investigations, this expert consensus reviews the definition, injury classification, rehabilitation assessment, rehabilitation strategies and rehabilitation measures of TSCI so as to provide early standardized rehabilitation treatment methods for TSCI.
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Objective To observe whether the transplanted dermal multipotent stem cells(dMSCs)transfected by adenovirus vector of CXCR4(Adv-CXCR4)can distribute more frequently to the wound of rats with combined wound and irradiation injury.Methods dMSCs transfected by Adv-CXCR4(group A),or transfected by adenovirus vector of green fluorescent protein(group B),and non-transfected dMSCs were labeled with 3H-TdR and then transplanted into combine-injured rats.The amount of dMSCs in wound were determined by liquid scintillation,and wounds healing process was observed by measuring the remaining wound area.Results From the 5th day after transplantation,the amount of dMSCs in the wound of group A accounted for 1.95%-3.85% of the total transplanted dMSCs,significantly greater than those in group B and group C,which accounted for 1.07%-1.86% of the total transplanted dMSCs.The remaining wound area in group A was smaller than those in group B and group C from day 12 after injury,and the healing time of group A was 1.5 day ahead than group B and group C.Conclusions dMSCs transfected by Adv-CXCR4 distributes more frequently to the wound of combine-injured rats and could accelerate wound healing.
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In this study, we prepared the acellular bone matrix of the inbred-line Banna mini-pig by using tissue engineering method and evaluated its possible application in bone tissue engineering. Histological analysis, xenoantigen expression and biomechanical measurement were performed on the matrix. HE staining and scanning electron microscopy showed the cellular components were almost removed. Immunohischemical result demonstrated that the xenoantigen, alpha-gal,was also eliminated. There was no statistically significant difference between the acellular bone matrix group and control group. The acellular bone matrix can provide appropriate space structure and strength for grafts. In conclusion, our data suggest that acellular bone matrix is a new kind of ideal bone scaffold material.
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Animals , Female , Male , Antigens, Heterophile , Biomechanical Phenomena , Bone Matrix , Allergy and Immunology , Stress, Mechanical , Swine , Swine, Miniature , Tissue Engineering , alpha-GalactosidaseABSTRACT
Cadherin-11 is a kind of classical cadherin subfamliy.The function of cadherin-11 involves embryonic development,tissue morphogenesis,tumor's invasion and metastasis,signal transduction and so on.This review summarizes the function of cadherin-11 in joint formation,including the relation of cadherin-11 with the bone and cartilage,growth plate,synovial and tendon formation.
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This study was aimed to examine the effectiveness of a gene transfer of human TGFbeta1 gene into endothelial cells and to determine whether TGFbeta1 increases ECM expression of endothelial cells. With the help of DOTAP, endothelial cells were transfected with pMAMneoTGFbeta1. The positive cell clones were selected with G418. The stable transfection and expression of TGFbeta1 in the endothelial cells were determined by immunofluorescence analysis. The expression levels of collagen I and fibronectin in the transfected and untransfected endothelial cells were determined by Western blot. The adhesion force between endothelial cells and matrix was determined by a micropipette technical system. The results showed abundant TGFbeta1 stable expression in the endothelial cells. TGFbeta1 gene was noted to increase collagen I and fibronectin expression and increase the adhesion between endothelial cells and matrix. These findings indicated that TGFbeta1 can be used in vascular tissue engineering for the enhancement of endothelial cells adhesion.
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Humans , Cell Adhesion , Cells, Cultured , Collagen Type I , Genetics , Endothelium, Vascular , Cell Biology , Physiology , Extracellular Matrix , Metabolism , Physiology , Fibronectins , Genetics , Tissue Engineering , Transforming Growth Factor beta , Genetics , Umbilical Veins , Cell BiologyABSTRACT
This study was conducted to examine the effectiveness of a gene transfer of human TGF beta 1 gene into smooth muscle cells and whether the TGF beta 1 can increase elastin expression of smooth muscle cells. With the help of DOTAP, smooth muscle cells were transfected with pMAMneoTGF beta 1. The positive cell clones were selected with G418. The stable transfection and expression of TGF beta 1 in the smooth muscle cells were determined by immunofluorescence analysis. The expression of elastin in the transfected and untransfected cells were determined by in situ hybridization. The adhesion force between smooth muscle cells and matrix was detected by micropipette system. The results showed abundant TGF beta 1 stable expression in smooth muscle cells. TGF beta 1 gene can increase two-three times elastin expression and increase the adhesion between smooth muscle cells and matrix. TGF beta 1 can be used in vascular tissue engineering to increase smooth muscle cells adhesion.
Subject(s)
Humans , Cell Adhesion , Cells, Cultured , Elastin , In Situ Hybridization , Muscle, Smooth, Vascular , Cell Biology , Metabolism , Transfection , Transforming Growth Factor beta , Genetics , Physiology , Transforming Growth Factor beta1ABSTRACT
Objective To detect whether mice bone marrow mesenchymal stem cells(BMSCs)can contribute to the regeneration of hepatocytes in bone marrow chimeric mice.Methods Female recipient mice(C57BL/6J)underwent whole body gamma-ray irradiation with a dose of 10 Gy to ablate their bone marrow,followed by immediate tail vein injection of BMSCs isolated from male GFP transgenic mice.Animals were killed at different phase points:1 week,1 month,and 3 months.Using fluorescence microscope we directly observed GFP-positive cells in the liver frozen sections,and we also prepared the parafilm sections to detect the GFP-positive cells and the coexpression of GFP and Alb,CK18 by immunohistochemistry and immunofluorescence respectively.Results We found numerous GFP-positive cells in recipient mice liver at 1 week after BMSCs transplantation,some at 1 month and seldom at 3 months.There were some cells coexpressing GFP and Alb,CK18 at all the phase points.Conclusion Allotype BMSCs can differentiate into Alb and CK18 positive hepatocytes in bone marrow chimeric mice,which will become an ideal cell resource for liver tissue project.
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Objective To further determine their possible synergistic effect on accelerating wound healing, adenovirus vector containing recombinant human hPDGF-A and hBD2 genes was constructed and the expression of exogenous genes in transformed mesenchymal stem cells derived from rat bone marrow was observed. Methods By putting IRES in the middle of hPDGF-A and hBD2, these two genes were expected to be expressed individually. The shuttle vector was named as pAdTrack-hPDGF-A-IRES2-hBD2, which homologously recombinated with Adeasy-1 in BJ5183 cells and formed the mammalian expression vector pAdeasy-hPDGF-A-IRES2-hBD2. Furthermore, the recombinant vector was packaged in 293 cells into infectious recombinant adenovirus, which were used to infect BMSCs. The expression of hPDGF-A and hBD2 in BMSCs was detected by RT-PCR. Results We successfully constructed recombinant adenovirus vector that simultaneously expressed hPDGF-A and hBD2. The expressions of hPDGF-A and hBD2 were confirmed by RT-PCR on transformed BMSCs. Conclusion The established BMSCs that overexpressed hPDGF-A and hBD2 provide a new strategy of combining cell therapy and gene therapy to promote wound healing, especially the chronic one.
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Objective To construct the recombinant adenovirus of mouse Osf2/Cbfal gene and to observe its ability to infect NIH3T3 fibroblasts. Methods The Osf2/Cbfal gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria. Then the recombinant adenovirus was transfected into NIH3T3 cells using Lipofectine DOTAP, The target gene was detected by poly-merase chain reaction (PCR). The titer and its infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. Results Restriction endonuclease and PCR analyses confirmed that the Osf2/Cbfal gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.6?1012 pfu/ml. The adenovirus had a strong effect on NIH3T3 cells. Conclusion The recombinant adenovirus containing Osf2/Cbfal gene was successfully constructed by the method of homogenous recombination in bacteria.
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Objective To explore the feasibility of mobilization circulating MSCs by G-CSF and observe the repairing effect of G-CSF mobilization in severe mouse traumatic brain injury(TBI) model.Methods MSCs-derived bone marrow and peripheral blood(PB) were cultured and its CFU-F were counted after mobilization by G-CSF.At 2,24,48,96,120,144,192,264,336 h after severe TBI in mice was establish,the neurobehavior of mice was measured by neurological examination and motor functional test,and mortality rate and pathologic changes were analyzed.Results MSCs-derived PB were successfully cultured.The CFU-F of mobilization group increased significantly than that of control group(P
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Objective To isolate the bone marrow mesenchymal stem cells(MSCs) from the GFP-expressing mouse, and to study the osteoblastic differentation of the cells. Methods MSCs were isolated by density gradient centrifugation, then the clutrued cells were induced to osteoblastic differentiation using the conditional medium. We detectd the expression of GFP and MSCs differentiation into osteoblasts by histochemistry and immunochemistry. Results The MSCs maintained the expression of GFP during expanded and induced process. After induced for 10 days, lots of alkaline phosphatase and osteoclcin staining positive cells were observed.Conclusion The MSCs of GFP-expressing mouse were successfully isolated and differentiated into ostoblasts. It may be valuable for tracing the seeding cells in tissue engineering bone.
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The development of body donation remains relatively slow in China.One of its main influencing factors restricted to the development of donation is the anatomy teachers' quality.Lots of volunteers to donate and their families are not satisfied with it,which eventually leads to the failure of the donation.Therefore,enhancing anatomy teachers' quality has become one of the key steps in the donated work.It needs to improve the idea awareness of teachers,to standardize the behavior of teachers,to enhance the role models of teachers,and to carry out a regular humanities education and training to truly realize "people-oriented" goals.
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Objective To study the biological characteristics of platelet-derived growth factor A and human beta defensin 2 (hPDGF-A/hBD2) gene-modified rat bone marrow mesenchymal stem cells (BMSCs). Methods By using liposome transfection technique, recombinant adenovirus vector expressing hPDGF-A/hBD2 (Adv-hPDGF-A-IRES-hBD2) labeled with GFP was transfected into 293T cells for virus packaging and amplification. BMSCs were isolated, cultured and infected by adenovirus-containing supernatant. The exogenous gene-modified BMSCs were comprehensively studied on their biological features, in terms of morphology, cell growth curve, cell cycle, and adipogenic, osteogenic and myogenic differentiation ability. Results hPDGF-A-IRES-hBD2 gene-modified BMSCs did not show obvious changes in cell viability, proliferation, cell cycle distribution or cell differentiation. Conclusion BMSCs were not only good carriers for exogenous hPDGF-A and hBD2 genes but also seed cells for cell therapy even after hPDGF-A/hBD2 modification.
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Objective To observe the repair effect of human ?-defensin 2 (hBD 2)-modified rat dermal multipotent stem cells (dMSCs) transplantation on infected wound. Methods Thirty Wistar rats were excised a piece of whole-layer back skin, 3 mm in diameter, then infected the wound with 1?10 8/ml pseudomonas aeruginosa 1 ml, then the rats were injected on the wounded back respectively with dMSCs modified by hBD 2 (n=10), or pure dMSCs (n=10) or none as control (n=10). The repair effect was evaluated by observing the amount of bacteria under the scar, wound healing time and the percentage of remaining wound area. Results The amount of bacteria under the scar in rats that were transplanted with dMSCs modified by hBD 2 was less than that in rats transplanted with dMSCs or controls (P
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Objective To examine the expression of human ?-defensin 2 (hBD_ 2 ) recombinant adenovirus expression vector in rat dermal multipotent stem cells (dMSCs) and to observe the antiseptic activity of recombinant hBD_ 2 . Methods The expression of hBD_ 2 in dMSCs was examined by RT-PR, fluorescent immunochemistry and Western blotting, and the concentration of recombinant hBD_ 2 in supernate was measured by ELISA. The antiseptic activity of recombinant hBD_ 2 was assessed by K-B disc agar diffusion test. Results hBD_ 2 could be effectively expressed in dMSCs, and the concentration of recombinant hBD_ 2 in supernate was about 743.6 ng/ml . Recombinant hBD_ 2 in supernate showed antiseptic activity. Conclusion Recombinant adenovirus expression vector of hBD_ 2 could be effectively expressed in dMSCs, and the recombinant hBD_ 2 in supernate showed obvious antiseptic effects toward some standard bacteria lines.