Revision of Schatzker type Ⅵ tibial plateau fracture failure focus on the recovery of lower limb alignment / 中华外科杂志

Objective@#To explore the influence of the lower extremity abnormal alignment and the joint surface, and to explore the surgical skills.@*Methods@#Twenty-two cases of tibial plateau Schatzker Ⅵ fracture internal fixation failure revision from January 2012 to January 2017 in Department of Orthopedics, Shanghai 10th Hospital.One year follow-up after initial surgery to make sure of failure.Three-dimensional CT scan, radiography, infection index, gait analysis, knee joint ROM, femur tibia angle, tibial plateau tibial shaft angle and posterior slope if tibial plateau were observed. The medial approach and bi-planer osteotoma were used.Autogenous iliac bone graft, postoperative fast recovery channel were used.Follow-up point included preoperative and postoperative 7 days, 6 weeks, 3 months, and 6 months.Obvervational index included double lower limbs radiography, knee society score(KSS), complications such as infection, skin necrosis, joint main passive activity, double lower limbs alignment the last follow-up SF-36 scale.Rate was compared by χ2 test, measurement data using paired sample t test.Correlation was analyzed by Pearson correlation regression testing.@*Results@#Twenty-two patients received follow-up.KSS, more than 21 cases were benign, with good gait.One case was poor, with claudication gait.Not skin necrosis, no deep infection cases, 1 case get blisters 2 days postoperatively, and disappear after 5 days with detumescence and cold therapy.Whether restoring force line affect the KSS significantly(χ2=22.000, P=0.000). Knee joint ROM, SF-36 score, KSS and lower limb alignment were improved significantly. In different individual the articular surface and anatomical angle recovered greatly but the posterior slope angle was quite difference which has no correlation with KSS and SF-36 scale(P>0.01).@*Conclusions@#Revision of Schatzker type Ⅵ tibial plateau fracture failure should focus on the recovery of lower limb alignment.moderate overcorrect bone cutting and joint surface height can bring benefits to the postoperative knee function.Revision surgery patients have greater psychological pressure, more early psychological intervention is necessary.

Explore the judgmental of the indications of total knee arthroplasty using deep convolutional neural network / 中华骨科杂志

Objective To explore the feasibility of the deep convolutional neural network (DCNN) judging the indications and prognosis of the total knee arthroplasty based on the trained DCNN computer learning system.Methods C1FAR-10 DCNN model based on TensorFlow (an open source system,Google,USA) optimized by Alex Krizhevsky were constructed.There were 400 cases with knee osteoarthritis from different databases used for analysis.Three hundred patients underwent total knee arthroplasty,while 100 did not.X-ray of 200 preoperative cases from the 400 cases and their enlarged image (50 times) were applied for training DCNN,while the enlarged images from other 200 cases were used to test the DCNN.The correlation and the regression between judgment of the DCNN and clinical truth were analyzed.The clinical truths were rechecked three times and were confirmed by treatment results.Pearson correlation and linear regression analysis were used.The relation test of the software was only used as a reference.Results There was no significant difference between the baseline of cases for learning and test.After learning 200 cases,the DCNN judged the 10 000 cases enlarged from remaining 200 cases.The correlation between the DCNN judgment and the clinical truth was not significant (r=0.000,F=0.001,P=0.970).False positive was observed in 1 681 cases,false negative in 3 296.After enlarged to 10 000 images,the correlation between the two judgments was significant (F=11 228.735,P=0.000,r=0.727 and R2=0.529).The software detection precision was 0.860.Conclusion DCNN can be applied in judging the indications of the total knee arthroplasty.Large sample size can improve the accuracy of the judgment significantly.

Lentiviral-medicated hβD-3 gene transfer induces chondrogenesis of human synovium derived mesenchymal stem cells and its antibacterial activity / 中华骨科杂志

Objective This article is aim to test the safety and the in vitro antibacterial activity of the Lentiviral-nedicated humanβ-defensin 3 (hβD-3) transfecion synovium-derived mesenchymal stem cells (SMSCs).Methods Tissues with SMSCs were obtained by surgical operations.Cells were explant cultured and purified by magnetic cell separation system.Cells were identified by microscopic observation, immunofluorescence and cell surface markers.Through inducing the cells into fat, osteoblasts and chondroblasts to respectively determine the multi-directional differentiation potential.Construct a hβD-3 contained lentivirus and transfected into SMSCs.Got the cell growth curve and determine the DNA content by using flow cytometry.The NOD/SCID mice were applied to verify the tumorigenicity of SMSCs.Agar diffusion test was applied to doing antibacterial activity test of posttransfecton SMSCs.The rabbit model of knee joint in staphylococcus aureus infections verify its bacteriostatic action in vivo.Results: Purified SMSCs had the structure and surface signatures of MSCs.It had the potential of multi-differentiation.Immunofluorescence had verified that SMSCs transfected by lentivirus could stably express hβD-3.Cell proliferation kinetics, karyotype analysis and Tumorigenic type analysis verified the safety of SMSCs after transfection.The in vivo and vitro antibacterial activity test of the recombinant SMSCs showed that after cell passages, the subcultured SMSCs (P5 cells were used in this article) could express hβD-3 stably and had antibacterial activity.The result of the antibacterial circle showed that low concentration group inhibit bacterial activity while the medium and high group could enlarge compared with the negative and positive control group.Conclusion Lentiviral-medicated hβD-3 gene express SMSCs could successfully subcultured and stably express hβD-3, meanwhile it had apparent in vitro antibacterial activity.

Experimental study on the compound culture of small intestinal submucosa with synovial mesenchymal stem cells / 中华骨科杂志

Objective To investigate the biosecurity and biocompatibility of small intestinal submucosa (SIS) as scaffold for tissue engineering and to explore the possibility to induce synovial mesenchymal stem cells (SMSCs) differentiate into cartilage with SIS as the scaffold and SMSCs as the seed cells. Methods The SMSCs were isolated and cultured from the synovial mem-brane of knee joints of rabbits by a conventional method. The SIS was harvested from pigs according to the physical method and Abraham's method. 4 rabbits are divided into the experimental group and control group. The biosecurity of SIS as scaf-folds were investigated in pyrogen test, skin sensitization test and systemic toxicity test. The SMSCs and SIS were co-cultured in vitro and induced to differentiate into cartilage to explore the biocompatibility of SMSCs and SIS, the proliferation and differ-entiation ability of SMSCs on the scaffold of SIS. The varietyies were identified by the microscope. Results The SIS isolated with the physical method and Abraham's method is a milky and translucent membrane with a smooth surface. Under the electron microscope, SIS presented a porous Stereoscopic three-dimensional network structure, which is formed by fibrous tissues' intertex-ture. Its' porosity was about 80%and its aperture was 100-200μm. Meanwhile, the biosecurity and biocompatibility of SIS were also fine. In the trial that the SMSCs and SIS were co-cultured in vitro, the SMSCs can grow, adhere to and differentiate on the sur-face of SIS and into the hollows very well. It can also secrete extracellular matrix. Through scanning microscope observation, cells contact with each other by their neuritis, or adhered to the wall of hole by cellular protrution. On the surface of SIS, the SMSCs maintain the property that it can easily differentiate into the chondrogenic lineage in the chondrogenic medium. Immunochemistry staining showed positive expression of type II collagen post 21 days of co-cultrue. Conclusion SIS can be used as the scaffold to construct tissue engineering meniscus as it has good biosecurity and biocompatibility with SMSCs without disturbing the cell form or inhibiting the growth and function of SMSCs.

Biological characteristics of synovial mesenchymal stem cells after isolation and purification / 中国组织工程研究

BACKGROUND:A large amount of studies have confirmed that synovial mesenchymal stem cells have the similarity in cellmorphology, immune phenotype, colony forming ability and differentiation potential with bone marrow mesenchymal stem cells. But bone marrow mesenchymal stem cells are better than synovial mesenchymal stem cells in the ability to differentiate into cartilages. OBJECTIVE:To discuss the possibility of using synovial mesenchymal stem cells as seed cells for meniscal tissue engineering. METHODS:The synovial mesenchymal stem cells were isolated from rabbit synovial tissues with limiting dilution monoclonal culture method, and then the cells were purified. The morphology, ultrastructure, molecular phenotype, proliferation kinetics, karyotype and tumorigenicity of the in vitro cultured cells were analyzed. RESULTS AND CONCLUSION:The synovial mesenchymal stem cells isolated from the rabbit synovial cells had high proliferation capacity during in vitro monolayer culture. The synovial mesenchymal stem cells grew to peak at 6 days, and the doubling time was (30.2±2.4) hours. Flow cytometry results showed the synovial mesenchymal stem cells could express some molecular makers of mesenchymal stem cells, such as CD44 and CD90. DNA contents check, karyotype test and oncogenicity test confirmed isolated and purified synovial mesenchymal stem cells were the normal diploid cells without tumorigenicity, so the cells can be used as seed cells for meniscal tissue engineering.

Growth factors in the construction of tissue-engineered meniscus / 中国组织工程研究

BACKGROUND:Differentiation of growth factor-induced cells into fibrocartilage is the hot topic in the research of meniscus tissue engineering. The in vitro construction and in vivo reconstruction of menisci are closely related with the effect of growth factors. OBJECTIVE:To review the current development of growth factors in meniscus tissue engineering, and to investigate the mechanism. METHODS:The VIP database (http://lib.cqvip.com), CNKI database (www.cnki.net) and PubMed database (http://www.ncbi.nlm.nih.gov/pubmed) were retrieved for the related articles from January 2008 to March 2013 with the key words of“meniscus tissue engineering, cartilage, growth factors”. A total of 53 articles on the research of growth factors in meniscus tissue engineering were included. RESULTS AND CONCLUSION:There were various growth factors in cartilage tissue engineering research, and the new growth factors have been found in the constant. The research on the regulation effect of growth factors on cartilage was changed from the research of single growth factor model to the research of the interactions between multiple growth factors;the molecular mechanisms of growth factors in regulating the cartilage have also been extensively studied. Growth factors have good application prospects in tissue engineering, but there are stil many problems to be solved, such as during the healing of menisci, the expressions and the effect of different growth factors are different in different time points. So how to play the interaction between growth factors timely and appropriately thus stimulating the in vivo microenvironment, investigating the molecular mechanisms of growth factors in regulating the cartilage, as wel as finding the new growth factors are the research focus in meniscus tissue engineering.

Influence to the curative effect of THA with different methods to reconstruct limb length for Crowe Ⅳ type developmental dysplasia of the hip / 中华骨科杂志

Objective To explore the differences of the joint function and the patient's satisfaction between two different methods to reconstruct limb length for unilateral Crowe Ⅳ developmental dysplasia of the hip (DDH).Methods The clinical controlled study were used.21 cases with unilateral DDH Crowe Ⅳ treated with total hip arthroplasty were divided into 2 groups randomly.The patients in the first group were reconstructed the limb with the compensatory length and those in the second group were did with the real length.The patients were followed up 10 years.Two independent sample t-test was used to compare these two operations.Harris scores and SF-36 scores were used in the test.Survival analysis with Kaplan-Meier method was involved in the survival of the prosthesis and then test with Log-rank test.X-ray films of different period were compared to confirm the prosthetic loosening.Results Seventeen patients were followed up 8-10 years.There were no significant differences both in Harris score and most of the SF-36 scores at the median follow-up of 10 years.The item mental healthy of SF-36 expressed significant difference.Five patients required revision.There were 10 cases suffered with polyethylene wear,6 cases with severe osteoporosis in greater trochanter,and 3 cases with osteolytic reaction.Conclusion There are no significant differences between the two surgical method in the survival rate of prosthesis and the joint function in the median follow up.The satisfaction of the control groups is lower than the trial groups both at the early stage after the operation and the median follow up.The revision rate between the two groups is similar but the reason is different.

Histocompatibility between small intestinal submucosa and synovial mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Smal intestinal submucosa is characterized as antimicrobial activity, good biocompatibility, bio-mechanical properties, and rapid degradation in vivo, similar to the extracellular matrix of meniscal fibrochondrocytes. OBJECTIVE:To observe whether there exists a good histocompatibility between smal intestinal submucosa and synovial mesenchymal stem cells. METHODS:Smal intestinal submucosa was treated with physical and chemical treatment. And hematoxylin-eosin staining and scanning electron microscopy observation were performed. Then, smal intestinal submucosa extracts were prepared for the fol owing experiments. (1) Pyrogenic test:smal intestinal submucosa extracts were injected into the ear vein of New Zealand white rabbits. (2) Skin sensitization test:smal intestinal submucosa extracts, paraformaldehyde solution and normal saline were respectively injected intradermal y into New Zealand white rabbits. (3) General toxicity test:smal intestinal submucosa extracts and normal saline were respectively injected into the ear vein of New Zealand white rabbits. Smal intestinal submucosa was co-cultured with osteogenic rabbit synovial mesenchymal stem cells, and smal intestinal submucosa cultured alone served as control. RESULTS AND CONCLUSION:There were some intestinal mucosal epithelial cells, fat cells and other cells adhered onto the surface of smal intestinal submucosa after physical treatment. While, the amount of residual cells decreased sharply after chemical treatment. But the main structure and the component had not been changed. The surface of smal intestinal submucosa was smooth and no cells remained, and there was a three-dimension network spatial structure. The porosity was 80%. Smal intestinal submucosa is a non-toxic, nonirritating, non-immunogenic biomaterial with very good biocompatibility, which has a good histocompatibility with rabbit synovial mesenchymal stem cells.