A study on the biological effects of electromagnetic pulse with different high⁃intensity on condylar cartilage in rats / 口腔疾病防治

Objective@#To explore the biological effects of electromagnetic pulse (EMP) with different high intensities on condylar cartilage in rats. @*Methods@#SD rats were randomly divided into a sham group (Sham) and an irradiation group (EMP1: 500 kV/m, 10 Hz; EMP2: 270 kV/m, 10 Hz). Then, they were sacrificed at 1 h, 3 h, 12 h, 24 h and 3 d after irradiation. The degree of cartilage degeneration was evaluated by HE, safranine O-fast green, type Ⅱ collagen immunohistochemistry and TUNEL staining. Immunohistochemistry and western blot were performed to detect the expression of the matrix degradation factors: matrix metalloproteinase-13 (MMP-13), a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS-5) and the apoptosis key factor cleaved-cysteinyl aspartate specific proteinase (cleaved-Caspase3) in condylar cartilage. @*Results @#HE staining showed that, compared with the Sham group, a small amount of exfoliation was found on the fibrous surface layer of the cartilage after irradiation in the EMP1 and EMP2 groups. Compared with the Sham group, the percentage of safranine O-fast green-positive area decreased significantly at 12 h and 24 h (both P<0.01) in the EMP1 group and 12 h and 24 h in the EMP2 group (both P<0.05); the percentage of type Ⅱ collagen-positive area decreased significantly at 3 h and 12 h (P<0.05, P<0.001) in the EMP1 group. In addition, the number of TUNEL-positive apoptotic cells increased significantly at 1 h, 3 h, 12 h, and 24 h in the EMP1 group and 1 h, 3 h, and 12 h in the EMP2 group (P<0.05). Moreover, at different timepoints (except at 3 d) in the EMP1 group and EMP2 group, the percentage of MMP-13, ADAMTS-5- and cleaved Caspase3-positive chondrocytes and their protein levels in condylar cartilage increased significantly after irradiation (P<0.05). @* Conclusion@# EMP with a certain degree of high-intensity can induce early transient damage to condylar cartilage. This effect is dose-and time-dependent.

Effect of Buyi Pishen acupuncture on inflammatory factor and cartilage matrix in adjuvant arthritis rats / 中国针灸

OBJECTIVE@#To observe the effect of Buyi Pishen acupuncture (acupuncture for invigorating spleen and kidney) on inflammatory factor and synovial cartilage matrix in adjuvant arthritis (AA) rats, and to explore the mechanism of acupuncture for rheumatoid arthritis (RA).@*METHODS@#A total of 60 clean male Wistar rats were randomized into a normal group, a model group, a tripterygium wilfordii polyglycoside tablet (TWP) group and an acupuncture group, 15 rats in each group. Rats in the model group, the TWP group and the acupuncture group received intradermal injection of Freund's complete adjuvant (FCA) at right hind foot pad to induce the AA model. TWP suspension of 8 mg/kg was given by gavage in the TWP group. Acupuncture was applied at "Shenshu" (BL 23), "Pishu" (BL 20) and right "Housanli" (ST 36), "Sanyinjiao" (SP 6), "Yanglingquan" (GB 34) in the acupuncture group, 15 min a time, once a day. The intervention was given 15 days in both TWP group and acupuncture group. The foot-pad swelling degree before modeling, before and after intervention and the arthritis index (AI) score before and after intervention were calculated; the serum levels of interleukin (IL)-1β, IL-4, IL-10 and tumor necrosis factor-α (TNF-α) were detected by ELISA method; the ultrastructure and histomorphological changes of synovium issue were observed by transmission electron microscope and HE staining; the positive expression of matrix metalloproteinase (MMP)-3 and MMP-9 in synovium issue was detected by immunohistochemistry method.@*RESULTS@#Before intervention, foot-pad swelling degree of the model group, the TWP group and the acupuncture group was increased compared with the normal group (P<0.01). After intervention, foot-pad swelling degree and AI score were increased compared with the normal group (P<0.01), foot-pad swelling degree and AI scores in the TWP group and the acupuncture group were lower than the model group (P<0.05), and those in the acupuncture group were decreased compared with the TWP group (P<0.05). The model group exhibited unclear nuclear membrane of synovial cells, chromatin pyknosis, massive inflammatory cell infiltration and hyperplasia in synovial tissue; the TWP group and the acupuncture group exhibited clear and smooth nuclear membrane of synovial cells, inapparent chromatin pyknosis, less inflammatory cell infiltration and hyperplasia in synovial tissue, the acupuncture group exhibited less matrix destruction as well. Compared with the normal group, serum levels of IL-1β and TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were increased (P<0.01), while serum levels of IL-4 and IL-10 were decreased (P<0.01) in the model group. Compared with the model group, serum levels of IL-1β and TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were decreased (P<0.05, P<0.01), while serum levels of IL-4 and IL-10 were increased (P<0.05) in the TWP group and the acupuncture group; compared with the TWP group, serum level of TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were decreased (P<0.05), while serum levels of IL-4 and IL-10 were increased (P<0.05) in the acupuncture group.@*CONCLUSION@#Buyi Pishen acupuncture can effectively improve the injury of articular cartilage in AA rats, its mechanism maybe related to reducing the inflammatory reaction in synovium and inhibiting the degradation of articular cartilage matrix.

Development of Printable Natural Cartilage Matrix Bioink for 3D Printing of Irregular Tissue Shape

The extracellular matrix (ECM) is known to provide instructive cues for cell attachment, proliferation, differentiation, and ultimately tissue regeneration. The use of decellularized ECM scaffolds for regenerative-medicine approaches is rapidly expanding. In this study, cartilage acellular matrix (CAM)-based bioink was developed to fabricate functional biomolecule-containing scaffolds. The CAM provides an adequate cartilage tissue–favorable environment for chondrogenic differentiation of cells. Conventional manufacturing techniques such as salt leaching, solvent casting, gas forming, and freeze drying when applied to CAM-based scaffolds cannot precisely control the scaffold geometry for mimicking tissue shape. As an alternative to the scaffold fabrication methods, 3D printing was recently introduced in the field of tissue engineering. 3D printing may better control the internal microstructure and external appearance because of the computer-assisted construction process. Hence, applications of the 3D printing technology to tissue engineering are rapidly proliferating. Therefore, printable ECM-based bioink should be developed for 3D structure stratification. The aim of this study was to develop printable natural CAM bioink for 3D printing of a tissue of irregular shape. Silk fibroin was chosen to support the printing of the CAM powder because it can be physically cross-linked and its viscosity can be easily controlled. The newly developed CAM-silk bioink was evaluated regarding printability, cell viability, and tissue differentiation. Moreover, we successfully demonstrated 3D printing of a cartilage-shaped scaffold using only this CAM-silk bioink. Future studies should assess the efficacy of in vivo implantation of 3D-printed cartilage-shaped scaffolds.

Bone regeneration with bone marrow mesenchymal stem cells, platelet rich plasma and decellularized cartilage matrices complex / 实用口腔医学杂志

Objective:To study the bone regeneration capacity of the complex of bone marrow mesenchymal stem cells(BMSCs) and platelet rich plasma (PRP) with decellularized cartilage matrix (DCM).Methods:BMSCs were isolated from young rabbit and cultured;PRP were prepared from the fresh blood of rabbit and the DCM were come from the fresh ears of rabbits and processed into a mixture of BMSCs-PRP-DCM.The mixture was injected subcutaneously into nude mice,8 weeks after injection bone regeneration was examine by HE staining and Massons staining decellularization.Results:The BMSCs show good differentiation capacity in vitro and the cartilage fragments were well decellularized.Excellent endochondral ossification ability of the complex was observed by in vivo experiment.Conclusion:The BMSCs-PRP-DCM complex has good capacity of endochondral ossification.

Curative Efficacy of Combined Use of Dexamethasone Palmitate, Vitamin B12 and Lidocaine in Treatment of Knee Osteoarthritis and Serum Levels of IL-1, MMP-3 and TIMP-1 / 现代生物医学进展

Objective:To investigate the curative efficacy of combined use of dexamethasone palmitate,vitamin B12 and lidocaine in the treatment of knee osteoarthritis and its effects on the seum levels of interleukin-1 (IL-1),matrix metalloproteinase-3 (MMP-3) and matrix metalloproteinase-1 (TIMP-1).Methods:84 patients with osteoarthritis of knee were selected from August 2014 to July 2015 in Sixth Affiliated Hospital of Xinjiang Medical University and divided into the observation group and the control group according to the order of admission,42 cases in each group.The observation group was given intra-articular injection of dexamethasone palmitate,vitamin B12 and lidocaine,and the control group was treated with traditional Chinese medicine.The clinical efficacy was evaluated after treatment.The changes of serum IL-1,MMP-3,TIMP-1 levels and visual analogue (VAS) score were compared before and at 1 months after treatment.Results:After treatment,the total effective rate of observation group was significantly higher than that of the control group (P＜0.05),the serum levels ofIL-1,MMP-3 levels and VAS score were significantly lower than those before treatment,serum TIMP-1 level was obviously higher than that before treatment(P ＜0.05),the serum levels of IL-1,MMP-3 levels were significantly lower in the observation group than those of the control group,while the serum TIMP-1 level was obviously higher than(P＜0.05) Conclusion:Intra-articular injection with dexamethasone palmitate,vitamin B12 and lidocaine could effectively decrease the levels of serum IL-1 and MMP-3,increase the level ofTIMP-1,and relieve the pain in the patients with knee osteoarthritis.

Research on matrix proteoglycan turnover of tissue engineering repaired cartilage / 西安交通大学学报(医学版)

Objective To examine matrix proteoglycan metabolic markers and probe into the turnover of matrix proteoglycan and enzyme-mediated role of matrix metalloproteinases (MMPs) and aggrecanases in reparative tissues with tissue engineering cartilage. Methods Tissue-engineered cartilage was constructed by cancellous bone matrix gelatin (BMG) with allogeneic chondrocytes in vitro for 2 weeks, then implanted to repair osteochondral defects of rabbit knee joint. Samples were obtained 6 months later to explore the expressions of 3-B-3(-) epitope, MMPs, MMP-generated epitope BC-4 and aggrecanases-generated epitope BC-13. Results In repaired tissues, the expression of 3-B-3(-) epitope increased, but that of MMPs and MMP-generated epitope BC-4 reduced. There was no expression of aggrecanases-generated epitope BC-13. Conclusion Expressions of 3-B-3(-), MMPs, BC-4 and BC-13 can help probe into the matrix proteoglycan turnover in reparative cartilage tissues. Anabolism exceeds catabolism in the repaired tissues. MMPs play an important role in the conservative baseline turnover of proteoglycan and remodeling of the graft tissues.

Significance of Magnetic Resonance Imaging for Early Diagnosis of Knee Ostarthritis(review) / 中国康复理论与实践

@#During the latest researches on knee ostarthritis,it is discovered that before the morphology changes of cartilage,there were changes of the content of proteoglycan and water and changes of alinement of surface layer collagen fibers in cartilage matrix.By the new technologies of DWI,proton density mapping image,23Na MRI,T2Mapping image and so on,not only the pathological changes of cartilage can be disclosed,the component changes of cartilage matrix can also be detected.Accompany with the progress in the technology of software and hardware,MRI will have more and more prospects in early diagnosis and therapeutic effect evaluation of knee ostarthritis.

Bone defect repair using allogenic ECM combined with chondrocytes in rabbits / 中国矫形外科杂志

[Objective]The characteristics and mechanism of the bone defect repair with the using allogenic extracellular cartilage matrix(ECM)were studied.[Method]Forty New Zealand white rubbits were divided into three groups at random:A and B on the right side,the defect was covered with the complex of ECM and autologous mesenchymal stem cells(MSCs),the ECM in the groups A and B,respectively.The defects on the left side served as controls correspondingly.The rabbits were killed at the 2nd,4th,6th,10th week.Samples were taken for radiological and histological studies.[Result]In the experimental sides the bone defects were healed.New bone appeared in the way of intramembranous ossification and entochondrostosis.[Conclusion]It is suggested that the complex of ECM and autologous MSCs has the effects on guiding bone regeneration and preventing from nonunion.