MiR-145-5p restrains chondrogenic differentiation of synovium-derived mesenchymal stem cells by suppressing TLR4.

Osteoarthritis (OA) is a progressive degeneration of articular cartilage with involvement of synovial membrane, and subchondral bone. Recently, cell-based therapies, including the application of stem cells such as mesenchymal stem cells (MSCs), have been introduced for restoration of the articular cartilage. Toll-like receptors (TLRs) were reported to participate in OA progression and MSC chondrogenesis. Here, the role and molecular mechanism of toll like receptor 4 (TLR4) in chondrogenic differentiation of synovium-derived MSCs (SMSCs) were investigated. Molecular markers (CD44, CD90, CD45 and CD14) on SMSC surfaces were identified by flow cytometry. Multi-potential differentiation capacities of SMSCs for chondrogenesis, adipogenesis and osteogenesis were examined by Alcian blue, oil red O and Alizarin red staining, respectively. TLR4 and miR-145-5p levels in SMSCs were assessed using RT-qPCR. The protein expression of TGFB3, Col II, SOX9 and Aggrecan in SMSCs was tested by western blotting. Cytokine secretions were analyzed with ELISA for IL-1ß and IL-6. Intracellular NAD+ content and NAD+/NADH ratio were assessed. The interaction between miR-145-5p and TLR4 was confirmed by RNA pulldown and luciferase reporter assays. In this study, SMSCs were identified to have immunophenotypic characteristics of MSCs. TLR4 knockdown inhibited chondrogenic and osteogenic differentiation of SMSCs. Mechanistically, TLR4 was targeted by miR-145-5p in SMSCs. Moreover, TLR4 elevation offset the inhibitory impact of miR-145-5p upregulation on chondrogenic differentiation of SMSCs. Overall, miR-145-5p restrains chondrogenesis of SMSCs by suppressing TLR4.

Bcl-xL mutant promotes cartilage differentiation of BMSCs by upregulating TGF-ß/BMP expression levels.

Bcl-xL is a transmembrane molecule in the mitochondria, with apoptosis-related and pro-metabolic functions, that also plays a role in chondrogenesis and differentiation. A Bcl-xL mutant, in which the GRI sequence is replaced by ELN, has no anti-apoptotic effect, while other biological functions of this mutant remain unchanged. The present study investigated the impact of this Bcl-xL mutant on cartilage differentiation and the expression levels of TGF-ß and bone morphogenetic protein (BMP). Human bone marrow mesenchymal stem cells (BMSCs) were transfected with Bcl-xL and Bcl-xL mutant (∆Bcl-xL) overexpression vectors. The cells were divided into four groups: Control (not subjected to any transfection), EV (empty pcDNA3.1-Bcl-xL vector), OV (Bcl-xL overexpression) and ∆OV (∆Bcl-xL overexpression). Saffron and toluidine blue staining was performed to observe cartilage tissue formation. Flow cytometry was conducted to measure BMSC apoptosis. The expression levels of TGF-ß and BMP were evaluated using reverse transcription-quantitative PCR (RT-qPCR) and western blotting. Compared with that in the control group, the expression levels of Bcl-xL in the OV group increased significantly (P<0.05). Western blotting and RT-qPCR results revealed that OV and ∆OV treatment increased the expression levels of TGF-ß and BMP in transfected cells, compared to their expression in the control and EV groups (P<0.05). Saffron and toluidine blue staining results showed that cartilage formation was increased in the ∆OV and ∆OV + Bax-/Bak-groups to similar degrees. Cell apoptosis in the ∆OV group did not change compared with that in the control group. The Bcl-xL mutant promoted cartilage differentiation of BMSCs and upregulated TGF-ß/BMP expression. This enhancement of chondrogenic differentiation was not related to the expression of Bax and Bak. Taken together, these findings provided for improved application of bone tissue engineering technology in the treatment of articular cartilage defects.

A New Individualized Three-Dimensional Printed Template for Lateral Ankle Ligament Reconstruction.

BACKGROUND Anatomical reconstruction using a semitendinosus tendon autograft is one of the most widely-used techniques for chronic lateral ankle instability (CLAI), and it can result in good biomechanical recovery for patients. The purpose of this study was to investigate the outcome of a novel individualized three-dimensional printed guide template for lateral ankle ligament reconstruction compared with the traditional surgical methods. MATERIAL AND METHODS We retrospectively studied 34 patients with CLAI who required lateral ankle ligament reconstruction. Patients were randomly divided into 2 cohorts: the template group (18 patients) and the conventional group (16 patients). The average operation duration and number of radiation exposures were compared between the 2 cohorts. The displacement of anterior talar and talar tilt angle were recorded at the last follow-up, and Karlsson-Peterson score and American Orthopedic Foot and Ankle Society Score (AOFAS) were also compared. RESULTS All patients had satisfactory ankle stability at the last follow-up. The average operation duration was 51.9±3.6 min and the average number of radiation exposures was 1.34±0.6 in the template group, and the average operation duration was 72.4±12.6 min and the average number of radiation exposures was 6.58±1.7 in the conventional group. Difference between the 2 cohorts was statistically significant. However, in AOFAS (95.2±2.5 vs. 94.9±2.2; P>0.01.) and Karlsson Score (94.7±3.6 vs. 93.8±4.1; P>0.01.), no significant differences were found between the 2 cohorts. CONCLUSIONS Both the template technique and the conventional method provided satisfactory outcomes for CLAI patients. However, the shorter operation duration and low number of radiation exposures in the template cohort suggest it is the better alternative for treatment of CLAI.

Use of Masquelet's technique for treating the first metatarsophalangeal joint in cases of gout combined with a massive bone defect.

BACKGROUND: To examine the safety and efficacy of Masquelet's technique as a surgical method for treating the first metatarsophalangeal joint in cases of gout accompanied by a massive bone defect. METHODS: From January 2010 to January 2016, eleven patients (7 males and 4 females; mean age 33.1 years; range, 23-43 years) received surgical treatment for a first metatarsophalangeal joint tophus which caused a serious bone defect. The first metatarsophalangeal bone defects ranged from 3-6cm, or nearly 50% of the length of normal bone. During the first stage of Masquelet's technique, we removed the tophus and infused that area with bone cement that contained antibiotics. Two months later, we performed the second stage, in which the prosthesis was replaced with iliac cancellous bone, and the operated area was stabilized via locking plate fixation. RESULTS: All of the surgeries were successful, and the 11 patients were followed up for an average of 10.9 months. Postoperative evaluations showed that 10 of the 11 patients healed between 9 and 14 days after the initial surgery. Bone fusion occurred between 2.3 and 3.6 months after the operation, and the average healing time was 3.0 months. One foot wound became infected, but healed after vacuum aspiration. When the American Association of Foot and Ankle Surgery Maryland Foot scoring system was used to evaluate the foot function of the 11 patients prior to surgery, all 11 patients were graded as "failures." Following surgery, 2 patients were graded excellent, 5 were good, 3 were fair, and only 1 patient failed. The total combined excellent and good rate was 63.6%. The total mean Maryland scores pre- and post-surgery were 27.8 points and 74.1 points, respectively; thus the average patient score increased by 46.3 points. CONCLUSIONS: Joints with advanced tophus nodules develop segmental bone defects. Masquelet's technique is an effective method for treating such nodules and their associated bone defects.

Surgical Treatment of Zone 1 Fifth Metatarsal Base Fractures Using the Locking Compression Plate Distal Ulna Hook Plate.

BACKGROUND: The distal ulna hook plate was recently introduced for the treatment of fifth metatarsal base fractures, but no special articles have reported the surgical results of the plate fixation of displaced or comminuted fifth metatarsal base fractures in zone 1, which are the most common forefoot fractures. METHODS: Twenty-one patients with Lawrence classification zone 1 fifth metatarsal base fractures with extra-articular displacement greater than 2 mm (n = 13) and intra-articular displacement fractures (n = 8) were included in the study. Patients were treated with the distal ulna hook plate. Patients were evaluated clinically and radiographically, and the functional outcomes were graded using the American Orthopaedic Foot and Ankle Society midfoot scoring system. Time to union and return to pre-injury levels of activity were calculated. RESULTS: Mean American Orthopaedic Foot and Ankle Society midfoot scores were 42.25 points (range, 27-55 points) preoperatively and 93.16 points (range, 87-100 points) 1 year postoperatively. Mean time to complete union was 61.9 days (range, 52-85 days). Nineteen patients reported returning to previous activities of daily living at a mean of 79.8 days (range, 59-91 days). Two patients showed radiographic signs of mild degenerative changes and noted mild pain, and one patient experienced hardware irritation. CONCLUSIONS: The distal ulna hook plate might be considered an effective surgical method for zone 1 displaced fifth metatarsal base fractures or multifragmentary, osteoporotic fifth metatarsal base fractures.

Membrane complement regulatory protein reduces the damage of transplanting autologous bone marrow mesenchymal stem cells by suppressing the activation of complement.

There are few studies on the interaction of transplanting autologous bone marrow mesenchymal stem cells (BMSCs) and complement. In order to further explore the effect of complement on BMSCs, BMSCs were obtained from bone marrow of 20 cases clinical patients, and then experimented in vitro. The cytotoxicity of complement on the mesenchymal stem cells in autologous human serum (AHS) was measured by Europium cytotoxicity assay. The complement membrane attack complex (MAC) deposited on the membrane surface was detected by flow cytometry. Finally, the cytotoxicity on BMSCs was measured after mCRPs overexpression or knockdown. We found that more than 90% of cells derived from bone marrow were identified to be mesenchymal stem cells through detection of cell membrane surface markers by flow cytometry. BMSCs harvested from the 20 patients all had cytotoxicity after incubated with AHS, and the cytotoxicity was significant higher than that incubated with complement inactivated autologous human serum (iAHS). Complement attack complex (MAC) could be detected on the BMSCs incubated with AHS, which implied the complement activation. We also found that mCRPs CD55 and CD59 overexpressions can resist the cytotoxicity induced by complement activation, while mCRPs CD55 and CD59 knockdown can enhance the cytotoxicity. Thus, the results indicated that mCRPs could effectively protect BMSCs from attacking by complement by suppressing the activation of complement.

Correlation of Adrenomedullin Concentrations with Knee Osteoarthritis Grade.

BACKGROUND Adrenomedullin, a recently identified myokine, has an anti-inflammatory effect. Therefore, we aimed to assess the correlation of adrenomedullin concentrations with the presence and grade of severity of knee osteoarthritis (OA). MATERIAL AND METHODS We recruited 187 knee OA patients and 109 healthy subjects. The severity of OA was evaluated using the Kellgren-Lawrence grading system. RESULTS Compared with the control group, the knee OA group revealed markedly higher adrenomedullin concentrations. Serum and synovial fluid (SF) adrenomedullin concentrations increased with increased KL grades. CONCLUSIONS Serum and SF adrenomedullin concentrations show a correlation with the severity of knee OA.

An Alternative Bundle-to-Bundle Suturing Technique for Repairing Fresh Achilles Tendon Rupture.

The main concern about conventional Achilles tendon repair surgical techniques is how to maintain the initial strength of the ruptured Achilles tendon through complicated suturing methods. The primary surgical problem lies in the properties of the soft tissue; the deterioration of the Achilles tendon, especially in its elasticity; and the surface lubricity of the local tissues. In the present study, we describe an innovative bundle-to-bundle suturing method that addresses these potential problems.

[Ectopic osteogenesis in vivo using bone morphogenetic protein-2 derived peptide loaded biodegradable hydrogel].

We investigated the development of an injectable, biodegradable hydrogel composite of poly(trimethylene carbonate)-F127-poly(trimethylene carbonate)(PTMC11-F127-PTMC11 )loaded with bone morphogenetic protein-2 (BMP-2) derived peptide P24 for ectopic bone formation in vivo and evaluated its release kinetics in vitro. Then we evaluated P24 peptide release kinetics from different concentration of PTMC11-F127-PTMC11 hydrogel in vitro using bicinchoninic acid (BCA)assay. P24/ PTMC11-F127-PTMC11 hydrogel was implanted into each rat's erector muscle of spine and ectopic bone formation of the implanted gel in vivo was detected by hematoxylin and eosin stain (HE). PTMC11-F127-PTMC11 hydrogel with concentration more than 20 percent showed sustained slow release for one month after the initial burst release. Bone trabeculae surround the P24/ PTMC11-F127-PTMC11 hydrogel was shown at the end of six weeks by hematoxylin and eosin stain. These results indicated that encapsulated bone morphogenetic protein (BMP-2) derived peptide P24 remained viable in vivo, thus suggesting the potential of PTMC11-F127-PT- MC11 composite hydrogels as part of a novel strategy for localized delivery of bioactive molecules.

Posterior malleolar fracture: technique and clinical experience of the posterolateral approach.

OBJECTIVE: To introduce the postero- lateral surgical approach to the posterior malleolar fracture and report its clinical outcomes in 32 cases. METHODS: This study consisted of 32 cases, 22 males and 10 females with the mean age of 48 years (range, 21-63 years), suffering from posterior malleolar fracture. All cases were treated with the posterolateral surgical approach to the ankle. The average follow-up period was 28 months (range, 24-35 months). The clinical outcomes of these cases were evaluated on the basis of the Olerud-Molander Ankle (OMA) score and plain radiographs. RESULTS: All cases showed radiological evidence of bony union at follow-up. The average OMA score was 82 points; 21 cases had excellent scores (90-100 points), 9 good (61-90 points), and 2 fair (31-60 points). The excellent-to-good rate was 93.8%. Although most cases did not show any wound dehiscence or necrosis, one patient had a superficial infection which healed after using antibiotic dressing and one had sural cutaneous nerve injury that underwent spontaneous remission without any treatment after three months. In addition, one presented with mild symptoms of peroneal tendonitis that disappeared after plate removal. CONCLUSION: The posterolateral approach offers an effective technique for fracture reduction and fixation of large posterior malleolar fragments.

[Clinical investigation of the minimal invasive osteotomy for the treatment of hallux valgus combined with tailor's bunion].

OBJECTIVE: To investigate the clinical effects of the minimal invasive osteotomy treatment for hallux valgus combined with tailor's bunion deformity. METHODS: From May 2007 to May 2009, Forty-nine feet of 32 patients (a mean age of 57 years old) of hallux valgus combined with tailor's bunion deformity were reviewed retrospectively, including 2 males(2 feet) and 30 females (47 feet). All patients were treated by the minimal invasive osteotomy. The axial and lateral films of all feet with loading were taken before and after operation. The hallux abducto valgus angle(HAV),intermetatarsal angle(IM), fourth-fifth intermetatarsal angle (IM 4 to 5), modified fourth-fifth intermetatarsal angle (MIM 4 to 5), Metatarsophalangeal-fifth angle (MPA) and lateral deviation of the fifth matatarsal angle (LDA) were compared before and after operation. Therapeutic effects of all the patients were observed after operation, the item including AFAS score and WEN Jian-min's therapeutic effects standard. RESULTS: The average operating time was 40 minutes. All the patients were followed up,and the duration ranged from 6 to 24 months. The HAV angle, IM angle, IM 4 to 5 angle, MIM 4 to 5 angle,MPA angle and LDA angle significantly decreased after operation. The AFAS score significantly raised after operation. According to the criteria, 33 feet got an excellent result, 15 good and 1 fair. CONCLUSION: The minimal invasive osteotomy treatment for hallux valgus combined with tailor's bunion is effective and safe. This method is mini-trauma,less pain and it can make a very soon rehabilitation. This method has widely clinical value.