An injectable thermosensitive hydrogel for sustained release of apelin-13 to enhance flap survival in rat random skin flap.

With the advantage of handy process, random pattern skin flaps are generally applied in limb reconstruction and wound repair. Apelin-13 is a discovered endogenous peptide, that has been shown to have potent multiple biological functions. Recently, thermosensitive gel-forming systems have gained increasing attention as wound dressings due to their advantages. In the present study, an apelin-13-loaded chitosan (CH)/ß-sodium glycerophosphate (ß-GP) hydrogel was developed for promoting random skin flap survival. Random skin flaps were created in 60 rats after which the animals were categorized to a control hydrogel group and an apelin-13 hydrogel group. The water content of the flap as well as the survival area were then measured 7 days post-surgery. Hematoxylin and eosin staining was used to evaluate the flap angiogenesis. Cell differentiation 34 (CD34) and vascular endothelial growth factor (VEGF) levels were detected by immunohistochemistry and Western blotting. Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were assessed by enzyme linked immunosorbent assays (ELISAs). Oxidative stress was estimated via the activity of tissue malondialdehyde (MDA) and superoxide dismutase (SOD). Our results showed that CH/ß-GP/apelin-13 hydrogel could not only reduce the tissue edema, but also improve the survival area of flap. CH/ß-GP/apelin-13 hydrogel also upregulated levels of VEGF protein and increased mean vessel densities. Furthermore, CH/ß-GP/apelin-13 hydrogel was shown to significantly inhibit the expression of TNF-α and IL-6, along with increasing the activity of SOD and suppressing the MDA content. Taken together, these results indicate that this CH/ß-GP/apelin-13 hydrogel may be a potential therapeutic way for random pattern skin flap.

Genome-wide identification of STATs and analysis of their role in sex determination in Pacific oysters (Crassostrea gigas).

STAT (signal transducer and activator of the transcription) proteins, are a group of highly conserved transcription factors and fundamental components of the JAK-STAT signaling pathway. They play crucial roles in a variety of biological processes, such as immunity, proliferation, differentiation, and growth. However, little information is known regarding their role in gonad development and sex determination in mollusks. In this study, we identified 3 STAT genes in Pacific Oyster Crassostrea gigas. Phylogenetic analysis showed that STATs from mollusks were highly conserved, and most of them had four identical motif regions, except for the STAT1 and STAT3 predicted sequences from Crassostrea hongkongensis. Tissue expression analysis indicated CgSTAT1 had a high expression level in most tissues, while CgSTAT3 had a low expression level in most tissues. Expression analysis of early developmental stages showed CgSTAT1 had a higher expression level from egg to D shaped larva and a lower expression level in subsequent stages. In contrast CgSTAT1, CgSTAT2 had a reverse expression pattern. Expression analysis of different developmental stages of diploid gonads indicated that CgSTAT1 had a higher expression level at the S1 and S3 stages relative to the S2 stage in females, while in males the S3 stage had a higher expression than than the S2 stage. The expression level of CgSTAT1 between diploids and triploids in females differed significantly, but there were no significant differences in males. Expression of CgSTAT2 differed significantly between diploid and triploid males. These data suggest an important role for STATs in sex differentiation in diploid and triploid oysters. Our study is the first to explore the role of STATs in sex differentiation and gonadal development in oysters, and will help us better understand the molecular mechanisms of sex differentiation in shellfish.

Genome-wide analysis of the cytochrome P450 gene family in Pacific oyster Crassostrea gigas and their expression profiles during gonad development.

The cytochrome P450 (CYP) gene superfamily plays a significant role in various physiological processes, producing different compounds such as hormones, fatty acids, and biomolecules. However, little information is known their roles during gonad development in Pacific oyster (Crassostrea gigas). In this study, total of 116 CgCYP (Crassostrea gigas cytochrome P450) genes were identified and their expression pattern was analyzed for the first time. The relative molecular weights of these CgCYP genes ranged from 63.52 to 113.41 kDa, and the length of encoded amino acids ranged from 103 to 993. And total 26 cis-acting elements of these CgCYP genes were identified. GO and KEGG enrichment analysis showed some CgCYP genes are essential for the metabolism of male and female sex hormones. Additionally, expression anslysis showed 69 CgCYP genes were over-expressed in early gonad development and triploid infertile individuals. More importantly, expression levels of CgCYP1, CgCYP15, CgCYP34, CgCYP46, CgCYP69, CgCYP87, CgCYP88, and CgCYP103, were found to be significantly higher in female gonad, suggesting their important roles in female gonad development. The results of this study will provide a better understanding of the CgCYP genes in the gonad development of Pacific oyster.

Characterization of MKK family genes and their responses to temperature stress and Vibrio parahaemolyticus infection in noble scallop Chlamys nobilis.

Mitogen-activated protein kinase kinase (MKK), the key element of the Mitogen-activated protein kinase (MAPK) signaling pathway, is crucial for the immune response to adverse environments in aquatic animals. Nevertheless, there is limited information regarding the role of the MKK gene family in mollusks. In our study, genome data and transcriptome were used to identify four MKK genes (CnMKK4, CnMKK5, CnMKK6, and CnMKK7) in the noble scallop. The result of the gene structure, motif analysis, and phylogenetic tree revealed that MKK genes are relatively conserved in bivalves. Moreover, four CnMKK genes were significantly highly expressed in immune-related tissues, suggesting that CnMKKs may related to bivalve immunity. Furthermore, CnMKK6 and CgMKK4 were significantly differentially expressed (P < 0.05) under 24 h of temperature stress, and all CnMKKs were significantly differentially expressed (P < 0.05) under 24 h of Vibrio parahaemolyticus infection. These results showed that the CnMKKs may have a significant impact under biotic and abiotic stresses. In conclusion, the result of the CnMKKs provides valuable insights into comprehending the function of MKK genes in mollusks.

Effects of chronic cold stress on tissue structure, antioxidant response, and key gene expression in the warm-water bivalve Chlamys nobilis.

As ectothermic invertebrates, mollusks are regarded as good environmental indicator species for determining the adverse effects of climate change on marine organisms. In the present study, the effects of cold stress on the tissue structure, antioxidant activity, and expression levels of genes were evaluated in the warm-water noble scallop Chlamys nobilis by simulating natural seawater cooled down during winter from 17 °C to 14 °C, 12 °C, 10 °C, and 9 °C. Firstly, the gill was severely damaged at 10 °C and 9 °C, indicating that it could be used as a visually indicative organ for monitoring cold stress. The methylenedioxyamphetamine (MDA) content significantly increased with the temperatures decreasing, meanwhile, the antioxidant enzyme activities superoxide dismutase (SOD) and catalase (CAT) showed a similar pattern, suggesting that the scallop made a positive response. More importantly, 6179 genes related to low temperatures were constructed in a module-gene clustering heat map including 10 modules. Furthermore, three gene modules about membrane lipid metabolism, amino acid metabolism, and molecular defense were identified. Finally, six key genes were verified, and HEATR1, HSP70B2, PI3K, and ATP6V1B were significantly upregulated, while WNT6 and SHMT were significantly downregulated under cold stress. This study provides a dynamic demonstration of the major gene pathways' response to various low-temperature stresses from a transcriptomic perspective. The findings shed light on how warm-water bivalves can tolerate cold stress and can help in breeding new strains of aquatic organisms with low-temperature resistance.

A novel carotenoids-producing marine bacterium from noble scallop Chlamys nobilis and antioxidant activities of its carotenoid compositions.

Marine bacteria produce many bioactive compounds, including carotenoids. However, the quality of bacterium carotenoids is relatively unknown. Therefore, in this study, a novel carotenoids-producing bacterium Brevundimonas scallop Zheng & Liu was isolated from Chlamys nobilis. The genome of the isolate was analyzed, carotenoid compounds were screened using HPLC-MS and the carotenoid production in B. scallop was monitored. The results revealed that the genome of B. scallop contained a carotenoid synthesis gene cluster, which involved in astaxanthin and hydroxy-astaxanthin biosynthesis. The 2,2'-dihydroxy-astaxanthin was the major carotenoid produced by B. scallop. The optimum culture condition for the highest carotenoids production (1303.62 ± 61.06 µg/g dry cells) for B. scallop was at temperature and salinity of 20 °C and 3% salt, respectively, in 10 g/L glucose as carbon source. The results showed the B. scallop is a new carotenoids resource in marine bivalve, which has an excellent antioxidative activity and potential industrial use.

Orthogonal plating method versus parallel plating method in the treatment of distal humerus fracture: A systematic review and meta-analysis.

PURPOSE: This systematic review and meta-analysis was performed to investigate the outcomes of orthogonal plating method and parallel plating method in the treatment of distal humerus fracture from the current literatures. METHODS: The electronic literature database of Pubmed, Embase, and Cochrane library were searched in November 2018. The data operation time, union time, Mayo Elbow Performance Score (MEPS), range of motion (ROM) of elbow, arc of elbow flexion, arc of elbow extension, rate of excellent and good results and complications (including heterotopic ossification, transient ulnar nerve neuropathy and ankylosis) were extracted. Stata 14.0 software was used for our meta-analysis. RESULTS: A total of 8 studies including 6 RCTs and 2 cohort studies met our inclusion criteria. This meta-analysis showed that there was no significant difference between the two groups regarding operation time, MEPS, ROM of elbow, arc of elbow flexion, arc of elbow extension and rate of excellent and good results at final follow-up (P = 0.50, P = 0.39, P = 0.87, P = 0.18, P = 0.58 and P = 0.59 respectively). However, the present meta-analysis demostrated that parallel plating method had significantly shorter union time than orthogonal plating method (P = 0.018). As for the complications (heterotopic ossification, transient ulnar nerve neuropathy and ankylosis), there was no significant difference between the two groups (P = 0.89, P = 0.08 and P = 0.29 respectively). CONCLUSION: Our meta-analysis suggested that both orthogonal plating and parallel plating method could achieve satisfactory outcomes with the similarly low complications in the treatment of distal humerus fracture. More RCTs are required for further research.

Intramedullary versus extramedullary fixation for the treatment of subtrochanteric fracture: A systematic review and meta-analysis.

PURPOSE: This meta-analysis was performed to investigate the outcomes of intramedullary fixation versus extramedullary fixation in the treatment of subtrochanteric fracture from the current literature. METHODS: The electronic literature database of PubMed, Embase, Cochrane library, CNKI and Wanfang were searched in December 2018. The data operation time, intraoperative blood loss, length of incision, length of stay, union time, rate of infection, rate of fixation failure, rate of refracture, rate of reoperation, rate of nonunion and rate of excellent and good results were extracted. Stata 14.0 software was used for our meta-analysis. RESULTS: A total of 11 studies including 8 RCTs and 3 cohort studies met our inclusion criteria. This meta-analysis showed that intramedullary fixation could achieve significantly shorter operation time (P = 0.000), less intraoperative blood loss (P = 0.000), shorter length of incision (P = 0.000) and length of stay (P = 0.001) with evidently lower rate of fixation failure (P = 0.001), rate of reoperation (P = 0.003) and higher rate of excellent and good functional results (P = 0.003) than extramedullary fixation for subtrochanteric fractures. However, no significant difference was found regarding union time (P = 0.17), rate of infection (P = 0.99), rate of refracture (P = 0.98) and rate of nonunion (P = 0.42) between the two groups. CONCLUSION: Our meta-analysis suggested that intramedullary fixation for subtrochanteric fracture might be superior to extramedullary fixation in term of shorter operation time, less intraoperative blood loss, shorter length of incision, length of stay and better functional outcomes. Meanwhile, intramedullary fixation had lower rate of fixation failure and reoperation. Therefore, we recommend intramedullary fixation as the treatment of subtrochanteric fracture. More large multi-center and high-quality RCTs are required for further research.

Correction to: Plumbagin Prevents IL-1ß-Induced Inflammatory Response in Human Osteoarthritis Chondrocytes and Prevents the Progression of Osteoarthritis in Mice.

The original version of this article contained mistakes, and the authors would like to correct them. The correct details are given below.

Inhibiting the PI3K/AKT/NF-κB signal pathway with nobiletin for attenuating the development of osteoarthritis: in vitro and in vivo studies.

Osteoarthritis (OA), an age-related degenerative disease, is characterized by progressive degradation of the articular cartilage. There is increasing evidence that nobiletin (NOB) exerts special biological functions in a variety of diseases. However, whether it protects against OA remains unknown. In this study, we investigated the anti-inflammatory and chondroprotective effects of NOB on IL-1ß-induced human OA chondrocytes and in the surgical DMM mice OA models. In vitro, NOB treatment completely suppressed the overproduction of pro-inflammatory mediators, including PGE2, NO, COX-2, iNOS, TNF-α and IL-6 in IL-1ß-induced human OA chondrocytes. Moreover, NOB exerted a potent inhibitory effect on the expression of MMP-13 and ADAMTS-5 as well as the degradation of aggrecan and collagen-II, which leads to the degradation of the extracellular matrix. Furthermore, NOB dramatically suppressed the IL-1ß-stimulated phosphorylation of PI3K/Akt and activation of NF-κB in human OA chondrocytes. In addition, treatment with NOB not only prevented the destruction of cartilage and the thickening of subchondral bone but also relieved synovitis in mice OA models. In conclusion, our study suggests that NOB holds novel therapeutic potential for the treatment of OA.

The protective effect of phloretin in osteoarthritis: an in vitro and in vivo study.

Osteoarthritis (OA) is a degenerative joint disease characterized by the degradation and inflammation of cartilage. Phloretin, a type of dihydrochalcone mainly found in apples and apple-derived products, has been reported to possess various potent biological effects such as antioxidant, anticancer, anti-inflammatory, and immunomodulatory. However, the anti-inflammatory effects of phloretin on OA have not been reported. This study was aimed at assessing the effects of phloretin on human OA chondrocytes. Human OA chondrocytes were pretreated with phloretin (10, 30, and 100 µM) for 2 h and subsequently stimulated with IL-1ß for 24 h. The production of NO, PGE2, TNF-α, and IL-6 was determined using the Griess reagent and ELISAs. The mRNA expression of COX-2, iNOS, MMP-3, MMP-13, and ADAMTS-5 was measured by real-time PCR. Changes in the protein expression of COX-2, iNOS, MMPs, ADAMTS, aggrecan, collagen-II, NF-κB, and the PI3K/Akt signaling pathway were detected by western blotting. In this study, we found that phloretin significantly inhibited the IL-1ß-induced production of NO, PGE2, TNF-α, and IL-6, the expression of COX-2, iNOS, MMP-3, MMP-13, and ADAMTS-5, and the degradation of aggrecan and collagen-II in human OA chondrocytes. Furthermore, phloretin dramatically suppressed the IL-1ß-stimulated phosphorylation of PI3K/Akt and activation of NF-κB in human OA chondrocytes. In addition, treatment with phloretin not only prevented the destruction of cartilage and the thickening of subchondral bone but also relieved synovitis in a mouse model of OA. Moreover, immunohistochemical results showed that phloretin significantly decreased the expression of MMP-13 and increased the expression of collagen-II in OA in mice. In conclusion, these results suggest that phloretin may be a potential agent for the treatment of OA.

Treatment of Die-Punch Fractures with 3D Printing Technology.

PURPOSE: We evaluated the feasibility, accuracy and effectiveness of applying three-dimensional (3D) printing technology for preoperative planning for die-punch fractures. METHODS: A total of 107 patients who underwent die-punch fracture surgery were enrolled in the study. They were randomly divided into two groups: 52 cases in the 3D model group and 55 cases in the routine group. A 3D digital model of each die-punch fracture was reconstructed in the 3D group. The 3D digital model was imported to a 3D printer to build the full solid model. The operation time, blood loss volume, and the number of intraoperative fluoroscopy were recorded. Follow-up was performed to evaluate the patients' surgical outcomes. RESULTS: Treatment of die-punch fractures using the 3D printing approach reduced the number of intraoperative fluoroscopy, blood loss volume, and operation time, but did not improve wrist function compared to those in the routine group. The patients wanted the doctor to use the 3D model to introduce the condition and operative plan because it was easier for them to understand. The orthopedic surgeons thought that the 3D model was useful for communicating with their patients, but their satisfaction with the preoperative plan was much lower than the benefit of using the 3D model to communicate with their patients. CONCLUSIONS: 3D printing technology produced more accurate morphometric information for orthopedists to provide personalized surgical planning and communicate better with their patients. However, it is difficult to use widely in the department of orthopedics.

The Feasibility of 3D Printing Technology on the Treatment of Pilon Fracture and Its Effect on Doctor-Patient Communication.

PURPOSE: The aim of this study was to assess the feasibility and effectiveness of the three-dimensional (3D) printing technology in the treatment of Pilon fractures. METHODS: 100 patients with Pilon fractures from March 2013 to December 2016 were enrolled in our study. They were divided randomly into 3D printing group (n = 50) and conventional group (n = 50). The 3D models were used to simulate the surgery and carry out the surgery according to plan in 3D printing group. Operation time, blood loss, fluoroscopy times, fracture union time, and fracture reduction as well as functional outcomes including VAS and AOFAS score and complications were recorded. To examine the feasibility of this approach, we invited surgeons and patients to complete questionnaires. RESULTS: 3D printing group showed significantly shorter operation time, less blood loss volume and fluoroscopy times, higher rate of anatomic reduction and rate of excellent and good outcome than conventional group (P < 0.001, P < 0.001, P < 0.001, P = 0.040, and P = 0.029, resp.). However, no significant difference was observed in complications between the two groups (P = 0.510). Furthermore, the questionnaire suggested that both surgeons and patients got high scores of overall satisfaction with the use of 3D printing models. CONCLUSION: Our study indicated that the use of 3D printing technology to treat Pilon fractures in clinical practice is feasible.

Comparison of suture button fixation and syndesmotic screw fixation in the treatment of distal tibiofibular syndesmosis injury: A systematic review and meta-analysis.

PURPOSE: This systematic review and meta-analysis was performed to investigate the outcomes of syndesmotic screw fixation versus suture button fixation in the treatment of distal tibiofibular syndesmosis injury from the current literature. METHODS: The electronic literature database of PubMed, Embase, and Cochrane library were searched in August 2018. The data on medial clear space, tibiofibular clear space, tibiofibular overlap, American Orthopaedic Foot and Ankle Society (AOFAS) scores and complications (including wound infection, local irritation or discomfort, screw loosening and screw breakage) were extracted. Stata 14.0 software was used for our meta-analysis. RESULTS: A total of 11 studies including 5 randomized controlled trials (RCTs) and 6 cohort studies met our inclusion criteria. This meta-analysis showed that there was no significant difference between the two groups regarding medial clear space (P = 0.54), tibiofibular clear space (P = 0.23) and tibiofibular overlap (P = 0.88) postoperatively. However, the present meta-analysis demonstrated that the suture button fixation group had significantly higher AOFAS scores than the syndesmotic screw fixation group at 3rd, 6th, 12th and 24th months postoperatively (P = 0.001, P = 0.006, P = 0.000 and P = 0.049 respectively). Besides, the time to full weight bearing in the suture button fixation group was significantly earlier than that in the syndesmotic screw fixation group (P = 0.000). As for the complications, the suture button fixation group had a lower rate of post-operative complication (screw loosening and screw breakage) compared with the syndesmotic screw fixation group (P = 0.048 and P = 0.000 respectively). CONCLUSION: Our meta-analysis suggested that suture button fixation could achieve significant higher AOFAS scores with a lower rate of postoperative complications and earlier time to full weight bearing in distal tibiofibular syndesmosis injury. More RCTs are required for further research.

Comparison of the outcomes of pediatric tibial shaft fractures treated by different types of orthopedists: A prospective cohort study.

BACKGROUND: The objective of this study was to compare the outcomes of pediatric tibial shaft fractures treated with titanium elastic nail (TEN) by pediatric orthopedists and non-pediatric orthopedists. MATERIALS AND METHODS: We conducted a prospective cohort study of 90 children of tibial shaft fractures, who were randomized to operative stabilization either by pediatric orthopedists (Group A, 45 cases) or by non-pediatric orthopedists (Group B, 45 cases) from April 2010 to May 2015. Demographic data and clinical characteristics (age, sex, weight, fracture side and type, cause of injury, number of fibula fracture and time from injury to operation) were comparable between the two groups before surgery. Clinical data, complications and functional outcomes between the two groups were recorded. RESULTS: A total of 10 patients were lost to follow-up for various reasons and each group remained 40 cases. The mean follow-up period was 14.9 ±â€¯1.9 months for Group A and 15.3 ±â€¯2.2 months for Group B (P = 0.451). There was no significant difference in length of hospitalization, full weight-bearing time, fracture union time and TEN outcome scores between the two groups (P = 0.917, P = 0.352, P = 0.404, P = 506, respectively). However, Group A exhibited significantly shorter operation duration and less fluoroscopy times than Group B (P < 0.001 and P < 0.001, respectively). Also, there was a trend for patients of Group A to have lower rate of open reduction than Group B (P = 0.019). When comparing the total complications, no significant difference existed between the groups (P = 0.764). CONCLUSIONS: Our results indicated that pediatric tibial shaft fractures treated surgically by pediatric orthopedists offered potential advantages including a shorter operating time, less times of fluoroscopy and a lower rate of open reduction. However, both pediatric and non-pediatric orthopedists could achieve satisfactory clinical results in treatment of these injuries.

Comparison of three surgical fixation methods for dual-bone forearm fractures in older children: A retrospective cohort study.

BACKGROUND: The aim of this study was to compare the outcomes of dual ESIN (D-ESIN) fixation, hybrid fixation, and open reduction and dual plate (d-plate) fixation in the treatment of dual-bone forearm fractures in children aged 10-16 years. MATERIALS AND METHODS: 137 patients with dual-bone forearm fractures (48 patients in the D-ESIN group, 45 patients in the hybrid group, and 44 patients in the d-plate group) were reviewed. Duration of surgery, length of incision, intraoperative blood loss, intraoperative times of fluoroscopy, and duration of postoperative immobilisation were recorded. Radiographic outcomes, functional outcomes, and complication rate were also recorded. RESULTS: Surgeries and incisions were significantly shorter, and less intraoperative blood loss occurred, in the hybrid group than the d-plate group (P < 0.001). The hybrid group was also characterised by less intraoperative fluoroscopy times and shorter duration of postoperative immobilisation compared with the D-ESIN group (P < 0.001). The union rate of the ulna at 3 months postoperatively was higher in the hybrid and d-plate groups than in the D-ESIN group (P = 0.003). The union rate of the radius was similar in all three groups (P = 0.403). No significant difference in the union rate of the radius or ulna was observed among groups at 6 months postoperatively (P = 0.052). The mean union time was notably later in the D-ESIN group than in the hybrid and d-plate groups. However, no significant difference in functional outcome or complication rate was observed among the three groups (P = 0.822 and P = 0.912). CONCLUSION: Hybrid fixation was superior in terms of the duration of surgery, intraoperative use of fluoroscopy, intraoperative blood loss, duration of postoperative immobilisation, delayed union of the ulna, and bone union time. Therefore, hybrid fixation is a safe and effective treatment for dual-bone forearm fractures in children aged 10-16 years.

Chrysin Attenuates IL-1ß-Induced Expression of Inflammatory Mediators by Suppressing NF-κB in Human Osteoarthritis Chondrocytes.

Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage degradation and inflammation. Chrysin, a natural flavonoid extracted from honey and propolis, has been reported to have anti-inflammatory effects. However, the anti-inflammatory effects of chrysin on OA have not been reported. This study aimed to assess the effects of chrysin on human OA chondrocytes. Human OA chondrocytes were pretreated with chrysin (1, 5, 10 µM) for 2 h and subsequently stimulated with IL-1ß for 24 h. Production of NO, PGE2, MMP-1, MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5 was evaluated by the Griess reaction and ELISAs. The messenger RNA (mRNA) expression of COX-2, iNOS, MMP-1, MMP-3, MMP-13, ADAMTS-4, ADAMTS-5, aggrecan, and collagen-II was measured by real-time PCR. The protein expression of COX-2, iNOS, p65, p-p65, IκB-α, and p-IκB-α was detected by Western blot. The protein expression of collagen-II and p65 nuclear translocation was evaluated by immunofluorescence. We found that chrysin significantly inhibited the IL-1ß-induced production of NO and PGE2; expression of COX-2, iNOS, MMP-1, MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5; and degradation of aggrecan and collagen-II. Furthermore, chrysin dramatically blocked IL-1ß-stimulated IκB-α degradation and NF-κB activation. Taken together, these results suggest that chrysin may be a potential agent in the treatment of OA.

Plumbagin Prevents IL-1ß-Induced Inflammatory Response in Human Osteoarthritis Chondrocytes and Prevents the Progression of Osteoarthritis in Mice.

Inflammation and inflammatory cytokines have been reported to play vital roles in the development of osteoarthritis (OA). Plumbagin, a quinonoid compound extracted from the roots of medicinal herbs of the Plumbago genus, has been reported to have anti-inflammatory effects. However, the anti-inflammatory effects of plumbagin on OA have not been reported. This study aimed to assess the effects of plumbagin on human OA chondrocytes and in a mouse model of OA induced by destabilization of the medial meniscus (DMM). In vitro, human OA chondrocytes were pretreated with plumbagin (2, 5, 10 µM) for 2 h and subsequently stimulated with IL-1ß for 24 h. Production of NO, PGE2, MMP-1, MMP-3, and MMP-13 was evaluated by the Griess reagent and ELISAs. The messenger RNA (mRNA) expression of COX-2, iNOS, MMP-1, MMP-3, MMP-13, aggrecan, and collagen-II was measured by real-time PCR. The protein expression of COX-2, iNOS, p65, p-p65, IκBα, and p-IκBα was detected by Western blot. The protein expression of collagen-II was evaluated by immunofluorescence. In vivo, the severity of OA was determined by histological analysis. We found that plumbagin significantly inhibited the IL-1ß-induced production of NO and PGE2; expression of COX-2, iNOS, MMP-1, MMP-3, and MMP-13; and degradation of aggrecan and collagen-II. Furthermore, plumbagin dramatically suppressed IL-1ß-stimulated NF-κB activation. In vivo, treatment of plumbagin not only prevented the destruction of cartilage and the thickening of subchondral bone but also relieved synovitis in mice OA models. Taken together, these results suggest that plumbagin may be a potential agent in the treatment of OA.

Silibinin protects against osteoarthritis through inhibiting the inflammatory response and cartilage matrix degradation in vitro and in vivo.

Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage degradation and inflammation. Silibinin, a polyphenolic flavonoid derived from fruits and seeds of Silybum marianum, has been reported to possess various potent beneficial biological effects, such as antioxidant, anti-cancer, hepatoprotective and anti-inflammatory activities. However, the anti-inflammatory effects of silibinin on OA have not been reported. This study aimed to assess the effects of silibinin on OA both in vitro and in vivo. In this study, we found that silibinin significantly inhibited the nterleukin-1ß (IL-1ß)-induced production of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α) and IL-6, expression of cyclooxygenase2 (COX-2), inducible nitric oxide synthase (iNOS), matrix metalloproteinase-1 (MMP-1), MMP-3, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) and ADAMTS-5, degradation of aggrecan and collagen-II in human OA chondrocytes. Furthermore, silibinin dramatically suppressed IL-1ß-stimulated phosphatidylinositol 3 kinase/ protein kinase B (PI3K/Akt) phosphorylation and nuclear factor-kappa B (NF-kB) activation in human OA chondrocytes. In addition, treatment of silibinin not only prevented the destruction of cartilage and the thickening of subchondral bone but also relieved synovitis in mice OA models. Also, the immunohistochemistry results showed that silibinin significantly decreased the expression of MMP-13 and ADAMTS-5 and increased the expression of collagen-II and aggrecan in mice OA. Taken together, these results suggest that silibinin may be a potential agent in the treatment of OA.

Baicalin suppresses IL-1ß-induced expression of inflammatory cytokines via blocking NF-κB in human osteoarthritis chondrocytes and shows protective effect in mice osteoarthritis models.

Osteoarthritis (OA) is a degenerative joint disease with an inflammatory component that drives the degradation of cartilage extracellular matrix. Baicalin, a predominant flavonoid isolated from the dry root of Scutellaria baicalensis Georgi, has been reported to have anti-inflammatory effects. However, the anti-inflammatory effects of baicalin on OA have not been reported. Our study aimed to investigate the effect of baicalin on OA both in vitro and in vivo. In vitro, human OA chondrocytes were pretreated with baicalin (10, 50, 100µM) for 2h and subsequently stimulated with IL-1ß for 24h. Production of NO and PGE2 were evaluated by the Griess reaction and ELISAs. The mRNA expression of COX-2, iNOS, MMP-3, MMP-13, ADAMTS-5, aggrecan and collagen-II were measured by real-time PCR. The protein expression of COX-2, iNOS, MMP-3, MMP-13, ADAMTS-5, p65, p-p65, IκBα and p-IκBα was detected by Western blot. The protein expression of collagen-II was evaluated by immunofluorescence. Luciferase activity assay was used to assess the relative activity of NF-kB. In vivo, the severity of OA was determined by histological analysis. We found that baicalin significantly inhibited the IL-1ß-induced production of NO and PGE2, expression of COX-2, iNOS, MMP-3, MMP-13 and ADAMTS-5 and degradation of aggrecan and collagen-II. Furthermore, baicalin dramatically suppressed IL-1ß-stimulated NF-κB activation. In vivo, treatment of baicalin not only prevented the destruction of cartilage but also relieved synovitis in mice OA models. Taken together, these results suggest that baicalin may be a potential agent in the treatment of OA.