BACKGROUND: Osteoarthritis (OA), the most common arthritis, is one of the most frequently encountered orthopaedic conditions. As a small number of large joints such as knee and hip are affected in OA, OA is an ideal target for local therapy. Although corticosteroid and hyaluronic acid have been traditionally used for joints through intra-articular (IA) injection, IA injection also provides a minimally invasive route to apply cell therapy to treat OA. IA cell therapy has drawn attention because it may provide regeneration of articular cartilage in addition to palliative anti-inflammatory effects. METHODS: Current progress of IA injection therapy and the author's perspective on this issue are described narratively. RESULTS: It is too premature to have any conclusion on the eventual efficacy of IA cell therapy concerning regeneration of articular cartilage based on current data. Prospective radiological and histological data from larger numbers of patients are needed to prove cost effectiveness of IA cell therapy. CONCLUSION: Expanding research in this field will produce further evidences to provide guidance on the eventual effectiveness of IA cell therapy in the future.
Arthritis ; Cartilage, Articular ; Cell- and Tissue-Based Therapy ; Cost-Benefit Analysis ; Hip ; Humans ; Hyaluronic Acid ; Injections, Intra-Articular ; Joints ; Knee ; Osteoarthritis ; Prospective Studies ; Regeneration

Objective: To elucidate the possible role of human lysozyme-like protein 4 (LYZL4) in fertilization and characterize its enzymatic properties. METHODS: The localization of LYZL4 in human spermatozoa was investigated by immunofluorescence staining, the sources of LYZL4 on the sperm surface examined by RT-PCR, and the role of LYZL4 in fertilization assessed by the zona-free hamster egg penetration test. The recombinant plasmid pPIC9K-LYZL4 was constructed and its expression induced with methanol after transformed into competent Pichia pastoris GS115. The recombinant LYZL4 protein (rLYZL4) was purified from the fermentation supernatant and subsequently identified by Western blot. The hyaluronan binding ability of rLYZL4 was determined by ELISA and the muramidase activity, hyaluronidase activity, and free radical scavenging ability examined by spectrophotometric methods. RESULTS: Immunodetection with a specific antiserum localized LYZL4 on the acrosomal membrane of mature spermatozoa, which was exclusively secreted from the testis and epididymis as shown by RT-PCR. Immunoneutralization of LYZL4 significantly decreased the number of human spermatozoa bound to zona-free hamster eggs in a dose-dependent manner in vitro. The recombinant protein was expressed successfully by the P. pastoris strain GS115. Purified rLYZL4 exhibited a potent hyaluronan binding ability and a strong free radical scavenging ability but no muramidase or hyaluronidase activity. CONCLUSIONS LYZL4 secreted from the testis and epididymis is localized on the acrosomal membrane of mature spermatozoa and plays a role in sperm-egg binding as well as in binding hyaluronan and scavenging free radicals, which suggests that it might be a multi-functional molecule contributive to sperm protection and sperm-egg binding.
Acrosome ; enzymology ; Animals ; Blotting, Western ; Cricetinae ; Enzyme-Linked Immunosorbent Assay ; Epididymis ; Female ; Fertilization ; physiology ; Free Radical Scavengers ; metabolism ; Humans ; Hyaluronic Acid ; metabolism ; Male ; Muramidase ; analysis ; physiology ; Pichia ; Plasmids ; metabolism ; Recombinant Proteins ; analysis ; metabolism ; Sperm-Ovum Interactions ; physiology ; Spermatozoa ; enzymology ; Testis

OBJECTIVETo investigate the effects of Dragon' s blood extract on proliferation and secret extracellular matrix function of fibroblasts in vitro.

METHODSDragon' s blood was extracted by chloroform, acetoacetic ester, alcohol. Human fibroblast were cultured in vitro in media containing gradient dilutions of Dragon' s blood extracts (0.002, 0.02, 0.2, 2, 20 mg/ml) , which was followed by cell proliferation assessed with MTT assay on 0, 12, 24, 36, 48, 60, 72 h. Under the optimal concentration, the cell growth curves were drawn and the flow cytometry (FCM) was used to determine the changes of cell cycle. On 0, 12, 24, 36, 48, 60, 72 h, the concentration of hyaluronic acid in the supernatant of fibroblast culture was measured by radioimmunoassay.

RESULTS0.2-2 mg/ml Dragon' s blood extracts enhanced the proliferation of fibroblasts in a dose-dependent manner. 2 mg/ml was the optimal dilution of Dragon's blood extract, and it increased the ratio of S cells in cell cycle [(25.80 ± 3.10)%] than control group [(7.50 ± 0.70)%, P < 0.01]. From 12 h to 72 h, in 2 mg/ml Dragon's blood group, concentration of Hyaluronic acid secreted by fibroblasts gradually increased, but were less than control (P < 0.01).

CONCLUSIONSDragon's blood acetoacetic ester extract improved the proliferation of cultured human fibroblasts in vitro, might be beneficial to promote wound healing.

Cell Cycle ; Cell Proliferation ; drug effects ; Culture Media ; chemistry ; Dose-Response Relationship, Drug ; Extracellular Matrix ; Fibroblasts ; cytology ; drug effects ; secretion ; Flow Cytometry ; Humans ; Hyaluronic Acid ; analysis ; secretion ; Plant Extracts ; pharmacology ; Resins, Plant ; Time Factors

OBJECTIVETo study the inhibitory effect of flavonoids from Glycyrrhiza uralensis on thioacetamide-induced chonic hepatic fibrosis in rats and the effect on the protein expressions of transforming growth factor-β1 (TGF-β1) and Caspase-3 in livers.

METHODMale Sprague-Dawley rats were randomly divided into totally seven groups: the normal control group, the model group, LF groups s (400, 200, 100, 50 mg · kg(-1) · d(-1)) and the silymarin positive control group (30 mg · kg(-1) · d(-1)). The hepatic fibrosis model was induced in the rats through intraperitoneal injection with 3% thioacetamide (TAA) at a dose of 150 mg · kg(-1) body weight twice a week for 12 weeks. During the course, the control group and the model group were orally administered with saline (1 mL · kg(-1) · d(-1)). After the modeling and drug intervention, the pathologic changes and fibrosis in liver tissues were observed by HE staining and Masson's Trichrome staining. The serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and liver hydroxyproline (HYP) contents were assayed by biochemical process. The serum hyaluronic acid (HA) was assessed by radioimmunoassay. In addition, the protein expressions of liver TGF-β1 and Caspase-3 were examined by immunohistochemical method. The mRNA expression of TGF-β1 in hepatic tissues was examined by quantitative Real-time PCR analysis.

RESULTCompared with the model group, flavonoids can protect the integrity of the structure of liver tissues, significantly reduce the hepatic cell degeneration and necrosis and the proliferation of fibrous tissues, notably reduce the serum AST, ALT, ALP and HA and HYP in hepatic tissues and down-regulate the protein expressions of liver TGF-β1 and Caspase-3 and the mRNA expression of TGF-β1 in hepatic tissues.

CONCLUSIONThe licorice flavonoids can resist the thioacetamide-induced hepatic fibrosis in rats. Its mechanism may be related to the down-regulation of the protein expressions of TGF-β1 and Caspase-3.

Animals ; Caspase 3 ; analysis ; Flavonoids ; pharmacology ; Glycyrrhiza uralensis ; chemistry ; Hyaluronic Acid ; blood ; Liver ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; prevention & control ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Thioacetamide ; Transforming Growth Factor beta1 ; analysis ; genetics

One hundred and five Labrador dogs were randomly divided into two groups to determine the number of animals that develop elbow dysplasia when treated with an oral supplement compared to untreated ones. Efficacy of the oral treatment was also evaluated once illness was diagnosed. The supplement (Hyaloral) contained hyaluronic acid, hydrolysed collagen, glucosamine, chondroitin sulphate, and gamma oryzanol. Clinical evaluation of the elbow joints was completed at months 3, 6, 12, and 20 by orthopaedic evaluations, radiography, serologic and blood analysis, and veterinarian evaluation of dysplasia symptoms. All side effects were recorded. In the control group, 33.3% of the dogs developed radiographic evidence of elbow dysplasia compared to 18.5% in the treated group. Symptoms of dysplasia at 12 months differed between the treated (12.5%) and control (61.5%) animals, and were significantly different at 20 months (p < 0.05). Differences in lameness along with movement and swelling of the elbows between groups were observed after 12 months. The treated group had improved significantly by the last visit (p < 0.05). No adverse side effects were reported. In conclusion, oral treatment with Hyaloral may have a potential cumulative action that provides protection against dysplasia and significantly improves symptoms of elbow dysplasia.
Administration, Oral ; Animals ; Collagen/*therapeutic use ; Dietary Supplements/analysis ; Dog Diseases/*prevention & control ; Dogs ; Drug Combinations ; Elbow Joint/*abnormalities ; Genetic Diseases, Inborn/prevention & control/*veterinary ; Hyaluronic Acid/*therapeutic use ; Joint Diseases/prevention & control/*veterinary ; Spain ; Species Specificity

Thirty-one dogs with patellar luxation (grades 2 and 3) were categorized into three groups. Group 1 (G.1; n = 12) had sodium hyaluronate (SHA) intra-articularly injected into the stifle joint that received surgery. Group 2 (G.2; n = 10) received SHA twice: first after surgery and then 1 week later. Group 3 (G.3; n = 9) served as a control, without injection. Blood was collected before injection and then once a week for 4 weeks after injection for evaluation of chondroitin sulfate (CS-WF6) and hyaluronan (HA). The results revealed significantly (p < 0.05) improved clinical scores by the end of week 4 in G.1 and G.2 relative to G.3; however, there was no significant difference between G.1 and G.2. There was a significant decrease (p < 0.05) in serum CS-WF6 levels beginning at week 2 in G.1 and G.2. At weeks 3 and 4, serum HA in G.1 and G.2 differed from that in G.3 (p < 0.05). No significant difference (p > 0.05) was observed in serum biomarkers between G.1 and G.2. In conclusion, intra-articular injection with SHA after joint surgery may improve homeostasis of the joint, retarding the process of OA.
Animals ; Blood Chemical Analysis/veterinary ; Chondroitin Sulfates/metabolism ; *Dogs ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Hyaluronic Acid/*administration & dosage/metabolism ; Injections, Intra-Articular/veterinary ; Male ; Osteoarthritis, Knee/drug therapy/prevention & control/*veterinary ; Stifle/*surgery ; Viscosupplements/*administration & dosage

Various kinds of glycosaminoglycans (GAGs) and proteoglycans (PGs) have been known to be involved in structural and space-filling functions, as well as many physiological regulations in skin. To investigate ultraviolet (UV) radiation-mediated regulation of GAGs and PGs in cultured human dermal fibroblasts, transcriptional changes of many types of PGs and GAG chain-synthesizing enzymes at 18 hr after 75 mJ/cm2 of UV irradiation were examined using quantitative real-time polymerase chain reaction methods. Hyaluronic acid synthase (HAS)-1, -2, and -3 and hyaluronidase-2 mRNA expressions were significantly increased by UV irradiation. Expressions of lumican, fibromodulin, osteoglycin, syndecan-2, perlecan, agrin, versican, decorin, and biglycan were significantly decreased by UV irradiation, while syndecan-1 was increased. Expressions of GAG chain-synthesizing glycosyltransferases, xylosyltransferase-1, beta1,3-glucuronyltransferase-1, beta1,4-galactosyltransferase-2, -4, exostosin-1, chondroitin polymerizing factor, and chondroitin sulfate synthase-3 were significantly reduced, whereas those of beta1,3-galactosyltransferase-6, beta1,4-galactosyltransferase-3, -7, beta-1,3-N-acetylglucosaminyltran sferase-2, and -7 were increased by UV irradiation. Heparanase-1 mRNA expression was increased, but that of heparanase-2 was reduced by UV irradiation. Time-course investigation of representative genes showed consistent results. In conclusion, UV irradiation may increase hyaluronic acid production through HAS induction, and decrease other GAG productions through downregulation of PG core proteins and GAG chain-synthesizing glycosyltransferases in cultured human dermal fibroblasts.
Cell Line ; Fibroblasts/metabolism/radiation effects ; Gene Expression Regulation/radiation effects ; Glucuronosyltransferase/genetics/radiation effects ; Glycosaminoglycans/*biosynthesis/chemistry ; Glycosyltransferases/genetics/*metabolism ; Humans ; Hyaluronic Acid/biosynthesis ; Hyaluronoglucosaminidase/genetics/radiation effects ; Polymerase Chain Reaction ; Proteoglycans/*biosynthesis/genetics/radiation effects ; RNA, Messenger/analysis/genetics ; Skin/*metabolism/radiation effects ; Transcription, Genetic/radiation effects ; *Ultraviolet Rays

The purpose of this prospective study was to verify and compare the strengths of various blood markers and fibrosis models in predicting significant liver fibrosis. One hundred fifty-eight patients with chronic liver disease who underwent liver biopsy were enrolled. The mean age was 41 yr and male patients accounted for 70.2%. The common causes of liver disease were hepatitis B (67.7%) and C (16.5%) and fatty liver (9.5%). Stages of liver fibrosis (F0-4) were assessed according to the Batts and Ludwig scoring system. Significant fibrosis was defined as > or =F2. Sixteen blood markers were measured along with liver biopsy, and estimates of hepatic fibrosis were calculated using various predictive models. Predictive accuracy was evaluated with a receiver-operating characteristics (ROC) curve. Liver biopsy revealed significant fibrosis in 106 cases (67.1%). On multivariate analysis, alpha2-macroglobulin, hyaluronic acid, and haptoglobin were found to be independently related to significant hepatic fibrosis. A new predictive model was constructed based on these variables, and its area under the ROC curve was 0.91 (95% confidence interval, 0.85-0.96). In conclusion, alpha2-macroglobulin, hyaluronic acid, and haptoglobin levels are independent predictors for significant hepatic fibrosis in chronic liver disease.
Adult ; Biological Markers/blood ; Chronic Disease ; Fatty Liver/complications ; Female ; Fibrosis ; Haptoglobins/analysis ; Hepatitis B/complications ; Hepatitis C/complications ; Humans ; Hyaluronic Acid/blood ; Liver Cirrhosis/complications/*diagnosis ; Liver Diseases/complications/*diagnosis ; Male ; Middle Aged ; Predictive Value of Tests ; Prospective Studies ; ROC Curve ; alpha-Macroglobulins/analysis

Agitation plays an important role in the hyaluronic acid (HA) fermentation process. However, views about the effect of agitation on HA production remain controversial. We investigated the effect of agitation on cell growth and HA synthesis during HA fermentation process by using Computational Fluid Dynamics (CFD) technology. The results showed that the biomass and HA yield changed a little with the increase of impeller speed, but the HA molecular weight firstly increased and then decreased. The results of phase agitation control strategy demonstrated that the influence of agitation on the HA molecular weight mainly exhibited at the stage of HA synthesis. Moreover, the CFD simulation results indicated that when impeller speed increased, the mixing time reduced while the shear rate increased significantly. The removal of anchor could moderate the contradiction between the mixing time and shear rate, and finally the HA molecular weight increased by 23.9%. The results of this work could provide guidelines for optimizing the HA fermentation, as well as the bioreactor design and scaling up.
Bioreactors ; Fermentation ; Hyaluronic Acid ; biosynthesis ; chemistry ; Hydrodynamics ; Numerical Analysis, Computer-Assisted ; Shear Strength ; Streptococcus equi ; growth & development ; metabolism

OBJECTIVETo investigate the clinical effects of moxibustion (Mox) in treating knee joint osteoarthritis, and to detect the change of hyaluronic acid (HA) level in serum and synovial fluid (SF) for evaluating its significance.

METHODSThirty OA patients were treated with Mox applied on inner and outer hsiyens and Ashi point for 10 min once a day, 5 times a week for 3 months and the therapeutic efficacy was evaluated. Serum and SF levels of HA were measured by radio-immunoassay before and after the 3-month treatment, and compared with those from 30 non-OA persons for normal control.

RESULTSAfter treatment, 19 patients (20 joints) out of the 30 patients were cured, the efficacy of Mox was evaluated as markedly effective in 8 patients on 10 joints, and as effective in 3 patients on 4 joints, the cure rate being 63.3%. Before treatment, HA level in serum (122.87 +/- 34.10 microg/L) was higher and in SF (0.98 +/- 0.17 g/L) was lower in OA patients than those in the normal controls (68.32 +/- 21.48 microg/L and 1.62 +/- 0.30 g/L, P<0.01), whereas after treatment, both the serum and SF levels of HA in patients changed toward normal range (70.29 +/- 27.30 microg/L and 1.58 +/- 0.26 g/L), showing insignificant difference as compared with those in the controls (P<0.01).

CONCLUSIONMox is an effective approach for treatment of OA, and the levels of HA in serum and SF can be taken as the quantitative indicators for evaluating the pathogenetic condition of OA patients.

Female ; Humans ; Hyaluronic Acid ; analysis ; blood ; Male ; Middle Aged ; Moxibustion ; Osteoarthritis, Knee ; blood ; metabolism ; therapy ; Synovial Fluid ; metabolism