1.Spinal fusion of lumbar intertransverse process in a goat by using tissue engineered bone with xenogeneic deproteinized bone as scaffold
Chunyang GAO ; Ge SUN ; Dongmei HAN
Orthopedic Journal of China 2006;0(11):-
[Objective]To study the properties of xenogeneic deproteinized bone (DPB) used as scaffold in the bone tissue engineering and its application to the spinal fusion of the lumbar intertransverse process in a goat. [Methods]The deproteinized bone was derived from an adult pig femoral cancellous bone through physical and chemical treatments.The cell-material complex was observed under the inverted phase contrast microscope to evaluate the adhesion and the growth of the osteoblasts. The experimental model of the spinal fusion in the lumbar intertransverse process was produced in 24 male goats aged 6-8 months, which were divided into 3 groups according to different implant graft. All the samples were harvested at 4,8,12 weeks postoperatively, and a series of examinations were performed, including the radiography and the histomorphological assay.[Results]It showed that The DPB maintained natural pore network system, it hardly had any antigen, so it had good histocompatibility. In the spinal fusion model of lumbar intertransverse process,the cell-material complex could form cartilage and had a new bone formation in a multipoint way, the osteogenic process was almost the same as the auto-ilium osteogenesis and had a good mechanical strength.[Conclusion]The xenogeneic deproteinized bone is a good material in the bone tissue engineering, which can be used as an osteogenesis scaffold and provides a stable fusion.
2.Construction of tissue-engineered bone using a xenogenic deproteinized bone scaffold for spinal intertransverse fusion
Gang LI ; Chunyang GAO ; Lianjin JIN
Chinese Journal of Tissue Engineering Research 2009;13(38):7459-7463
BACKGROUND:Xenogenic deproteinized bone is generally collected and has unique biological properties;however,its immunogen hopes be resolved so as to discover a new type of bone graft material.OBJECTIVE:To explore the performance of xenogenic deproteinized bone scaffold for tissue-engineered bone and the effect on spinal intertransverse fusion in goats.DESIGN,TIME AND SETTING:An in vivo cell-scaffold study was performed at the Key Laboratory of Fibrotic Biotherapy of Heilongjiang Province between February and October 2008.MATERIALS:Twelve healthy male goats aged 6-8 months were provided by Animal Center of Mudanjiang Medical College.METHODS:Cancellous bone at distal femur of adult swine was obtained to prepare xenogenic deproteinized bone scaffold using physical and chemical methods.Effects of the scaffold on morphology,structure,component,biomechanical property,and biological behavior of seed cells were detected and analyzed.Bone marrow was extracted from ilium of goat and gradient-centrifuged to obtain the third-passaged bone marrow mesenchyme stern cells (BMSCs).A certain quantity of autologous BMSCs and recombinant human bone morphogenetic protein 2 (rhBMP-2) were plated onto the scaffold to achieve tissue-engineeried bone.Models of bilateral L3,4 intertransverse bone graft were established on 12 goats.Tissue-engineeried bone was implanted into the left side in a repairing group,while the same volume of autologous ilium was implanted into the right side in a control group.MAIN OUTCOME MEASURES:X-ray examination and histological detection were performed at 4,8,and 12 weeks after implantation.RESULTS:Deproteinization-treated cancellous bone exhibited a spatial grid structure of variously sized,crossing and opening pores,composing hydroxyapatite and collagen.The pore diameter was 200-500 μm,and porosity was about 60%.Mechanical property and cell compatibility were well.X-ray examination demonstrated that at 4 weeks after implantation,some or even lateral intertransverse bridging regions were unclear,and material density in the repairing group was lower than control group;at 8 weeks after implantation,interspace between up and down bridging was shrunk,and a great quantity of calluses were successively formed;at 12 weeks after implantation,complete confluence was observed,and the density in the repairing group was closed to that in the control group.Histological detection indicated that at 4 weeks after implantation,new bone was multidrop-formed in the repairing group;at 8 weeks after implantation,bone tissue like the islet shape was passed through the whole implanted materials;at 12 weeks after implantation,woven bones arranged across,medullary cavity was formed,and osteogenic activity in the repairing group was closed to that in the control group.On the other hand,at 4 weeks after implantation,a large quantity of new bones were formed in the control group;at 8 weeks after implantation,a great quantity of collagen fibers were observed,and osteogenesis was clear in marginal region;at 12 weeks after implantation,fibrous tissues were reduced,and osteogenesis was active.CONCLUSION:BMSCs and rhBMP-2 incubated on xenogenic deproteinized cancellous bone is an ideal tissue-engineeried bone scaffold,characterizing by an excellent histocompatibility and a strong osteogenesis,based on in vitro X-ray diffraction analysis,mechanical test,and in vivo osteogenesis experiment.
3.Development and related characterization of heterogeneous bone scaffold materials
Chunyang GAO ; Ge SUN ; Dongmei HAN
Chinese Journal of Tissue Engineering Research 2007;0(03):-
BACKGROUND:Heterogeneous deproteinized bone because of wide source and its special biological characteristics maybe a good bone tissue engineering scaffold.Its immunogenicity and mechanical properties are different by different interventions.OBJECTIVE:To proof the preparation technology,immunological and mechanical properties of heterogeneous deproteinized bone.DESIGN,TIME AND SETTING:A control observational experiment was performed in the Heilongjiang Key Laboratory of Anti-fibrosis Biotherapy from February to October 2008.MATERIALS:Fresh femoral cancellous bones extracted from adult pigs,which were not including cartilages and cortical bones,were prepared into size of 3 cm?0.5 cm?0.5 cm.Their long axis direction was the same with orientation of trabeculae.The apparent pore densities of the used bones were almost same.METHODS:The bones were soaked in chloroform-carbinol(1:1) 24 hours for degrease.They were shaken and washed by distilled water at 50 ℃,and then put into 20% H2O2 for 24 hours.The procedure repeated three times.The bones were soaked in distilled water at room temperature for 24 hours,and then dryed.MAIN OUTCOME MEASURES:The gross morphological and histological features of the deproteinized cancellous bones were observed.The contents of amino acid and their biomechanical properties were measured.RESULTS:The three-dimensional space structures of the deproteinized cancellous bones were not damage greatly,and they had a natural pore network system which composited of hydroxyapatite and collagen.The contents of collagen amino acids in the deproteinized cancellous bones had no obvious difference from fresh cancellous bones,but the wave crests of aromatic amino acids such as tyrosine and methionine were disappear.The elastic moduli of deproteinized cancellous bones were significantly higher than those of fresh cancellous bones(P
4.Physicochemical properties of xenogeneic deproteinized cancellous bone as stent materials
Ge SUN ; Chunyang GAO ; Bin ZHOU
Chinese Journal of Tissue Engineering Research 2007;0(29):-
BACKGROUND:Deproteinized cancellous bone can be used as a new kind of bone graft material due to its wide resources andbiological characteristics if the problem of immunogen can be solved. OBJECTIVE:To study the properties of deproteinized cancellous bone following preparation with physico-chemical methods,in addition,to explore the feasibility of xenogeneic deproteinized cancellous bone used as scaffolds in bone tissue engineering. DESIGN,TIME AND SETTING:A comparison observation was performed at the Heilongjiang Key Laboratory of Anti-fibrosis Biotherapy between February 2008 and January 2009. MATERIALS:Deproteinized bone was derived from pigs and goals femoral cancellous bone followed by treating with 1:1 methanol extract and 30% H2O2. METHODS:The mechanical properties of vector were examined by scanning electron-microscopy,X-rays diffraction analysis,differential scanning calorimetry,and mechanical experimental instrument. The density of third passage of human bone mesenchymal stem cells (MSCs) of iliac bone was regulated to 2?108/L,vaccinated to vector,and the cell adhesion and proliferation was observed by inverted microscope and scanning electron microscope. Forty BALB/C mice were randomly dividedinto the experimental and control groups,with 20 animals in each group. Muscle behind femur was made muscle pouch,which was implanted pig deproteinized bone in the experimental group,and no other treatment in the control group. The level of serumcarnine,glutamic-oxal(o)acetic transaminase (GOT),as well as glutamic-pyruvic transaminase (GPT) were measured at weeks 12,4,8 after operation. Meanwhile,the immunogenicity of pig deproteinized bone and specimen of deproteinized bone in mice were detected at the 4 weeks after implantation. MAIN OUTCOME MEASURES:Physicochemical properties and immunogenicity of deproteinized bone were analyzed by in vitroexperiment. And the bio-safety was evaluated by animal experiment. RESULTS:The inorganic composition of deproteinized cancellous bone was hydroxyapatite ceramic,which is identical with nature composition of human bone. It showed that deproteinized cancellous bone maintained natural network pore system and kept good mechanical property. The MSCs grew well on deproteinized cancellous bone due to its low immunogenicity and toxicity. CONCLUSION:The xenogeneic deproteinized cancellous bone is an excellent vector in bone tissue engineering.
5.Expression of cytokines in the degenerative cartilages and synovial tissues of osteoarthritis patients
Yanhui ZHANG ; Chunyang GAO ; Shaohua LI
Chinese Journal of Tissue Engineering Research 2013;(37):6671-6675
BACKGROUND:The inflammation of osteoarthritis is mediated by chondrocytes and the synovial tissue-secreted cytokines. Articular cartilage and synovial tissue contains a variety of cytokines, which play important regulatory role in the repair of articular cartilage injury.
OBJECTIVE:To analyze the relationship between chondrocytes, synovial tissue-secreted cytokines and osteoarthritis, as wel as the effect of chondrocytes and synovial tissue-secreted cytokines on osteoarthritis.
METHODS:A computer-based search was conducted in Wanfang database (www.wanfangdata.com.cn), PubMed database (www.ncbi.nlm.nih.guv/pubmed) by the first author from 2005 to 2010. The key words were“osteoarthritis, degeneration, cartilage tissue, cytokines”. A total of 146 articles were obtained through computer search, then reading the titles and abstracts for initial screen, 86 articles with the unrelated research purposes were eliminated, 40 articles with the repetitive content were excluded, and final y 21 articles related to the effect of various cytokines in the degenerative cartilage and synovial tissue of the osteoarthritis patients were included for further analysis.
RESULTS AND CONCLUSION:The cytokines mainly refers to the activated immune cel s and some stromal cel-secreted non-specific regulate immune responses and inflammatory responses-mediated smal molecule proteins, including lymphokines produced by the lymphocytes, as wel as the single nuclear factor and other cytokines produced by monocytes macrophages. Synovial cel-secreted cytokines may partial y explain the pathological process of osteoarthritis and play an important role in joint inflammation. Although, more and more scholars have pay attention to the effect of synovial cel s and cartilage cel-secreted cytokines, but mainly studies the effect of exogenous cytokines on chondrocytes or synovial cel s, and the effect of endogenous cytokines in the pathogenesis of osteoarthritis has not been extensively studied.
6.Biocompatibility of induced bone mesenchymal stem cells on xenogeneic deproteinized cancellous bone
Gang LI ; Chunyang GAO ; Lianjin JIN
Chinese Journal of Tissue Engineering Research 2007;0(12):-
0.05). The cell cycles of the two groups were similar, and heteroploid cells were not found out in both groups. CONCLUSION: Xenogeneic deproteinized cancellous bone has a good biocompatibility to bone mesenchymal stem cells and promotes cell growth and differentiation.
7.ROI-CTM cervical fusion or titanium plate for treatment of Hangman fractures:a comparison of cervical stability and fusion rate
Weihong ZHANG ; Chunyang MENG ; Feng GAO ; Wen YUAN
Chinese Journal of Tissue Engineering Research 2015;(38):6074-6080
BACKGROUND:The self-locking ROI-CTM interbody fusion has zero profile design which can significantly reduce the impact on the esophagus and pharynx, and also simplify the operation procedure. However, there is no report whether it has satisfactory outcomes in the treatment of Hangman fractures.
OBJECTIVE: To compare the clinical effects of ROI-CTM or titanium plate internal fixation on postoperative cervical stability and fusion rate in the treatment of Hangman fractures.
METHODS:Twenty-one cases of Hangman fractures were included, including 13 males and 8 females, aged 25-62 years old. Fifteen cases were treated with titanium plate internal fixation, and the other five cases were treated with ROI-CTM fusion device. Cervical anteroposterior and lateral radiographs, lateral flexion extension radiographs and CT scanning were carried out before, 3 days and 3 months after repair to assess angular displacement, horizontal displacement and fusion rate; visual analog scale score and Bazaz dysphagia score were determined.
RESULTS AND CONCLUSION:At 3 days after operation, the ROI-CTM group was significantly better than the titanium plate group in visual analog scale scores and Bazaz dysphagia grade (P < 0.05), and there were no significant differences in angular displacement and horizontal displacement (P > 0.05). However, at 3 months after operation, there was no significant difference between the two groups in visual analog scale score, Bazaz dysphagia grade, angular displacement and horizontal displacement and fusion rate (P > 0.05). Postoperative complete reduction rate of spondylolisthesis was higher in the ROI-CTM group than the titanium plate group (P < 0.05). In each group, the angular displacement and horizontal displacement were significantly reduced at 3 day and 3 months after operation (P < 0.05), but there was no significant difference at 3 days and 3 months after operation (P > 0.05). No rejection reaction, internal fixation fracture and loosening occurred in the two groups. These findings indicate that the self-locking ROI-CTM fusion for treatment of Hangman fractures are characterized as higher reduction rate of spondylolisthesis, less dysphagia, and less neck pain.
8.Clinical significance of Her2 pathological expression in breast and gastric cancer patients
Zhenyu YAN ; Chunyang MAI ; Peng GAO ; Bo SU ; Shixuan WANG
Chinese Journal of Immunology 2016;32(6):858-862
Objective:To investigate the clinical significance of Her2 pathological expression in the breast and gastric cancer patients for providing a reference for cancer prevention. Methods: Selected surgical resection specimens of 60 diagnosed in advanced gastric cancer and 60 diagnosed with advanced breast cancer from March 2009 to May 2014 in our hospital,all specimens were given the Her2 pathological expression of immunohistochemical analysis and investigation the clinical and pathological data. Results: The Her2 positive expression rates in the breast cancer and gastric cancer specimens were 40. 0% and 36. 7%,respectively that compared to no significant difference (P>0. 05). The Her2 expression was not related to the cancer patient’s age,histological type,and there were sig-nificantly correlated to the TNM stage and lymph node metastasis ( P<0. 05 ) . Spearman correlation analysis showed that Her2 expression in the breast cancer and gastric cancer were significant positive correlation to the Survivin, Bcl-2 expression ( P<0. 05 ) . Conclusion:Breast cancer and gastric cancer patients have shown high pathological expression of Her2,and are related to the TNM stage and lymph node metastasis, it may be through inhibited the apoptosis regulating proteins involved in the pathogenesis and metastasis of tumors.
9.Characteristics of locking reconstruction titanium plate in the treatment of acetabular fracture for aged patients
Haibin WANG ; Chunyang MENG ; Bin WU ; Liang HAN ; Ming GAO ; Cunling JIA ; Dailiang JIA
Chinese Journal of Tissue Engineering Research 2014;(31):4962-4967
BACKGROUND:Pelvic and acetabular fractures in elderly are frequently observed in the clinic, and the incidence gradual y increased. Moreover, it is difficult to conduct reduction and fixation due to their physical status and osteoporosis to different degrees. Locking plate is ideal fixation material in surgical treatment of acetabular fractures in elderly due to the special functions and good molding of screw and locking plate.
OBJECTIVE:To analyze the advantages and disadvantages of locking reconstruction plate used in the acetabular fracture for the aged patients and to improve the clinical effect of acetabular fractures in elderly.
METHODS:From January 2010 to February 2013, 21 aged patients with acetabular fracture were treated by open reduction and internal fixation with locking reconstruction plates made of titanium and 00Cr 18 Ni 14 Mo 3 stainless steel in the Department of Orthopedics Affiliated Hospital of Jining Medical University.
RESULTS AND CONCLUSION:Of 21 patients, 20 cases were fol owed up for 12-18 months (mean 14 months), but one dropped out. Al the fractures were healed within 4-6 months and no failure cases. According to Matta evaluation criteria, the satisfactory rate was 90%. According to the D’Aubigne scoring system, the excellent and good rate was 95%. These data indicated that locking reconstruction titanium plate is reliable, has low failure rate and satisfactory clinical effects for acetabular fracture in aged patients, and is ideal fixation material in treatment of acetabular fractures in elderly.
10.Early curative effect of transpedicular screw fixation in treatment of thoracolumbar fracture combined with posterior ligamentous complex injuries
Guijun CAO ; Chunyang MENG ; Weihong ZHANG ; Xiangqing KONG ; Feng GAO ; Tongshen WU ; Guowu CHEN
Chinese Journal of Trauma 2015;31(4):307-311
Objective To investigate the preliminary clinical result of thoracolumbar fracture combined with posterior ligamentous complex injuries repaired by posterior transpedicular screw fixation.Methods A retrospective review was performed on 22 patients with thoracolumbar flexion-distraction fracture combined with posterior ligamentous complex injuries treated with transpedicular screw fixation from July 2008 to March 2013.There were 16 males and 6 females with mean age of 39 years (range,23-62 years).After medically stable,posterior pedicle screw fixation was performed under intravenousinhalational anesthesia.According to the degree of fracture displacement and types of ligament injury,posterolateral bone grafting or intervertebral fusion at the level of injury was conducted.Vertebral height restoration,Cobb' s angle and American Spinal Injury Association (ASIA) score were reviewed preoperatively,at postoperative 3 days and at the last follow-up.Results All the patients were operated on smoothly.There were no complications during operation.All the patients were followed up for 5-51 months (mean,26.5 months).Fracture reductions were satisfied with the closure of vertebral posterior element.Mean anterior vertebral height and Cobb' s angle improved by 20.6% and 10.60°respectively after operation (P <0.01).Eight patients with neurological dysfunction showed some recovery after operation with the mean sensory score improved by 20.7% (P < 0.05) and mean motor function score improved by 30.9% (P < 0.0l).All bone grafts were healed,without pain,loosening or breakage in the fixation system.Conclusions Posterior pedicular screw fixation attains good short-term outcome for thoracolumbar flexion-distraction fracture combined with posterior ligamentous complex injuries.The surgery provides satisfactory reduction and instant spinal three-column stability for the unstable spine fracture.Sufficient bone graft is the guarantee to permanent stability.