This is a report of attempts to compare the growth yields of various species of fastidious mycobacterium inch1ding human pathogens and non-pathogens in the conventional Dubos liquid medium and two simple media formulated recently; one is a medium containing 0.1% hyaluronic acid and 6.0% bovine serum albumin and the other is a semisyntheic medium made of umbilical cord extract supplemented with 10% sheep serum as a final concentration. All mycobacterial strains employed in experiments gave the heaviest growth yields in the hyaluronic acid-bovine serum albumin medium (HAS medium), among the three media. Dubos liquid medium seemed to be inferior to a medium made of umbilical cord extract (UCE medium) in supporting mycobacterial growth. There were three-to seven-fold increases in dry weight of the bacteria grown in the HAS medium as compared with those in the Dubos liquid medium. We also looked for the possible effect of bovine serum albumin (BSA)in the HAS medium on mycobacterial growth. As a result, we found that the amount of BSA in the HAS medium, ranging from zero to 6.0% in the medium, showed no substantial effect on the mycobacteria1 growth.
Comparative Study ; Culture Media/standards* ; Female ; Human ; Hyaluronic Acid/isolation & purification ; Hyaluronic Acid/pharmacology* ; Mycobacterium/growth & development* ; Pregnancy ; Tissue Extracts* ; Umbilical Cord*

Intra-articular injection of hyaluronic acid for osteoarthritis is an old and useful method. According to recent literatures, the following four problems were reviewed in this article: 1) Mechanisms. Real-time PCR was used to detect the transforming growth factor-beta 1 (TGF-beta1), vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) gene expression, which were significantly increased, as well as promoted secretion and synthesis of the tissue inhibitors of metalloproteinase, thus the reaction and cartilage destruction were decreased. 2) Methods. Because of discovering of new HA derivants, single intra-articular treatment with 6 ml hylan G-F 20 had replaced the routine method of once a week. To improve the therapeutic effectiveness, compound drags were applied, such as NSAIDs. 3) Indication. Knee OA (K-L II-III type) was first elected. But at present, the K-L I type was advocated early application. While the K-L IV type, although joint function can not improve, the symptoms relieved. 4) Therapeutic effectiveness. The double-blind place bo-controlled study showed that it was better than corticoid, and persistent and no side-effect. Therefore, this was a safe, persistent effective method.
Humans ; Hyaluronic Acid ; administration & dosage ; pharmacology ; therapeutic use ; Injections, Intra-Articular ; Osteoarthritis ; drug therapy

Hyaluronan (HA), a natural glycoaminoglycan featuring an extracellular matrix, has been suggested as an effective biocompatible material. In this study, the effectiveness of HA microparticles as a carrier system for antibiotics was evaluated, and their physicochemical characteristics were determined. Microparticles were fabricated by the gelation of sulfadiazine (SD) loaded HA solution with calcium chloride through either a granulation (GR-microparticles) or encapsulation (EN-microparticles) process, and atelocollagen was incorporated into the microparticles as an additive in order to improve their physical properties. The characteristics of the microparticles were examined by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and swelling test. In vitro release experiments were performed for 7 days and the released amount of SD was determined using high-performance liquid chromatography (HPLC). Microscopic observations revealed that the collagen incorporated HA particles had a more compact surface than the HA particles. DSC analysis determined a loss of SD crystallinity in the particles. Calcium chloride retarded the swelling of particles, whereas the loaded drug contents did not affect this property. Both GR-and EN-microparticles sustained SD release with initial bursting effect. SD release from EN-microparticles was faster than from GR- microparticles. In addition, the release rate was dependent on the SD content in the microparticles. These results suggest that collagen modified HA microparticles have a potential as a release rate controlling material for crystalline drugs such as SD.
Antibiotics/*administration & dosage ; Calcium Chloride/pharmacology ; Collagen/*pharmacology ; *Drug Carriers ; Hyaluronic Acid/*administration & dosage ; Sulfadiazine/administration & dosage

OBJECTIVE: To investigate the feasibility of a dual-crosslinked injectable hydrogel derived from acellular musclar matrix (AMM) for promoting myoblasts proliferation and myogenic differentiation. METHODS: Firstly, hyaluronic acid was oxidized with NaIO ₄ and methylated to prepare methacrylamidated oxidized hyaluronic acid (MOHA). Then, AMM obtained by washing enzymatically treated muscle tissue was aminolyzed to prepare aminated AMM (AAMM). MOHA hydrogel and AAMM were crosslinked using Schiff based reaction and UV radiation to prepare a dual-crosslinked MOHA/AAMM injectable hydrogel. Fourier transform infrared spectroscopy (FTIR) was used to characterize MOHA, AAMM, and MOHA/AAMM hydrogels. The injectability of MOHA/AAMM hydrogel were evaluated by manual injection, and the gelation performance was assessed by UV crosslinking. The rheological properties and Young's modulus of the hydrogel were examined through mechanical tests. The degradation rate of the hydrogel was assessed by immersing it in PBS. The active components of the hydrogel were verified using immunofluorescence staining and ELISA assay kits. The promotion of cell proliferation by the hydrogel was tested using live/dead staining and cell counting kit 8 (CCK-8) assays after co-culturing with C2C12 myoblasts for 9 days. The effect of the hydrogel on myogenic differentiation was evaluated by immunofluorescence staining and real time quantitative polymerase chain reaction (RT-qPCR). RESULTS: FTIR spectra confirmed the successful preparation of MOHA/AAMM hydrogel. The hydrogel exhibited good injectability and gelation ability. Compared to MOHA hydrogel, MOHA/AAMM hydrogel exhibited higher viscosity and Young's modulus, a reduced degradation rate, and contained a higher amount of collagen (including collagen type Ⅰ and collagen type Ⅲ) as well as bioactive factors (including epidermal growth factor, fibroblast growth factor 2, vascular endothelial growth factor, and insulin-like growth factor 1). The live/dead cell staining and CCK-8 assay indicated that with prolonged incubation time, there was a significant increase in viable cells and a decrease in dead cells in the C2C12 myoblasts within the MOHA/AAMM hydrogel. Compared with MOHA hydrogel, the difference was significant at each time point ( P<0.05). Immunofluorescence staining and RT-qPCR analysis demonstrated that the deposition of IGF-1 and expression levels of myogenic-related genes (including Myogenin, Troponin T, and myosin heavy chain) in the MOHA/AAMM group were significantly higher than those in the MOHA group ( P<0.05). CONCLUSION The MOHA/AAMM hydrogel prepared based on AMM can promote myoblasts proliferation and myogenic differentiation, providing a novel dual-crosslinked injectable hydrogel for muscle tissue engineering.
Hydrogels ; Hyaluronic Acid/pharmacology* ; Vascular Endothelial Growth Factor A/metabolism* ; Tissue Engineering/methods* ; Cell Differentiation ; Myoblasts/metabolism* ; Cell Proliferation

The purpose of this research is to construct a kind of 3D-Scaffold with galactose-carrying polysaccharide for improving the function of hepatocytes in vitro. Galactose moieties were covalently coupled with hyaluronic acid through ethylenediamine. Galactosylated hyaluronic acid/chitosan scaffolds were prepared by lyophilization. The characteristics of the scaffolds such as morphology, hydrophilicity, and mechanical properties were investigated. The results indicated that the porosity and the pore size of the scaffolds made in -20 degrees C were useful used for culturing hepatocytes. And, the incorporating of hyaluronic acid in chitosan network improved the hydrophilicity and mechanical properties of the scaffolds. Rat primary hepatocytes growing in the scaffolds observed by phase-contrast microscope showed the multicellular spheroid morphologies. Therefore, galactosylated hyaluronic acid/chitosan scaffolds could be used as a promising scaffold for liver tissue engineering.
Animals ; Cells, Cultured ; Chitosan ; chemistry ; pharmacology ; Galactose ; chemistry ; pharmacology ; Hepatocytes ; physiology ; ultrastructure ; Hyaluronic Acid ; chemistry ; pharmacology ; Liver ; physiology ; ultrastructure ; Porosity ; Rats ; Tissue Engineering ; Tissue Scaffolds ; chemistry

We compared the bone healing capacity of three different demineralized bone matrix (DBM) products applied using different carrier molecules (hyaluronic acid [HA] vs. carboxymethylcellulose [CMC]) or bone compositions (cortical bone vs. cortical bone and cancellous bone) in a rabbit segmental defect model. Overall, 15-mm segmental defects in the left and right radiuses were created in 36 New Zealand White rabbits and filled with HA-based demineralized cortical bone matrix (DBX), CMC-based demineralized cortical bone matrix (DB) or CMC-based demineralized cortical bone with cancellous bone (NDDB), and the wound area was evaluated at 4, 8, and 12 weeks post-implantation. DBX showed significantly lower radiopacity, bone volume fraction, and bone mineral density than DB and NDDB before implantation. However, bone healing score, bone volume fraction, bone mineral density, and residual bone area at 4, 8, and 12 weeks post-implantation revealed no significant differences in bone healing capacity. Overall, three DBM products with different carrier molecules or bone compositions showed similar bone healing capacity.
Animals ; Bone Matrix/*physiology ; Bone Transplantation ; Carboxymethylcellulose Sodium/*pharmacology ; Histology ; Hyaluronic Acid/*pharmacology ; Rabbits ; *Wound Healing ; X-Ray Microtomography ; X-Rays

OBJECTIVETo investigate the feasibility of using cartilage link protein of hyaluronic acid (HA-CLP) for defining the tumor boundary in a mouse model of lung carcinoma.

METHODSLung carcinoma was induced in KM mice by chemical carcinogenesis. HA-CLP separated from bovine cartilage and purified by affinity chromatography was labeled with (125)I for autoradiography. Immunohistochemical analysis and Western blotting were used to examine the efficiency of HA-CLP in defining the boundaries of the lung tumors.

RESULTSWith autoradiography, the clearest image of lung cancer was obtained at 2 h. With immunohistochemical method, the tumor boundary was the most clearly displayed at 2 h when the strongest signals of HA-CLP was detected; Western blotting also showed the clearest bands of HA-CLP at 2 h.

CONCLUSIONHA-CLP has the immunogenicity of HABP, and can efficiently indicate lung tumor boundary in autoradiography and immunohistochemistry.

Animals ; Autoradiography ; methods ; Extracellular Matrix Proteins ; metabolism ; pharmacology ; Female ; Hyaluronic Acid ; metabolism ; Immunohistochemistry ; Iodine Radioisotopes ; Lung Neoplasms ; radiotherapy ; Male ; Mice ; Proteoglycans ; metabolism ; pharmacology ; Radiotherapy, Image-Guided ; methods

AIMTo evaluate how solution viscosity affects the precorneal residence of five water-soluble polymers with different properties.

METHODSCaptive bubble technique was used, with the consecutive change of contact angle interpreted as an indication of desorption process, to study the residence of those polymers in vitro on freshly enucleated rabbit eyes under physiological conditions.

RESULTSCarbopol and sodium hyaluronate (HA), which adsorbed to isolated ocular surface more than 15 min, showed the optimum precorneal retentive capabilities. When the solution viscosity increased from 12 mPa.s to 50 mPa.s, the residence time of carbopol and HA were prolonged 10 min and 7 min, respectively, but that of sodium carboxymethylcellulose was not affected.

CONCLUSIONThe result suggested that higher viscosity is beneficial to improve the ocular residence time of bio-adhesive polymers.

Acrylic Resins ; Adhesiveness ; Animals ; Cornea ; drug effects ; metabolism ; Delayed-Action Preparations ; Drug Carriers ; Female ; Hyaluronic Acid ; pharmacokinetics ; pharmacology ; In Vitro Techniques ; Male ; Polyvinyls ; pharmacokinetics ; pharmacology ; Rabbits ; Solutions ; Viscosity

We have shown that hyaluronic acid stimulates the proliferation of quiescent NIH 3T3 cells. We have shown that treatment of 1 mg/ml hyaluronic acid results in increase of tyrosine phosphorylation of two proteins, MW 124 kDa and 60 kDa as detected by anti-tyrosine antibodies by Western blot analysis. Maximum phosphorylation occurred within 2 h after addition of 1 mg/ml hyaluronic acid. Stimulation of proliferation was also accompanied by increase in c-Myc protein, which was inhibited by amlloride, an inhibitor of Na+/H+ antiporter and EGTA and increase in the steady state level of pRb, the RB gene product. These results suggest that the intracellular signal transduction pathways that mediate the stimulatory effects of hyaluronic acid on cellular proliferation are similar to those of growth factors.
3T3 Cells ; Amiloride/pharmacology ; Animal ; Cell Division ; Dose-Response Relationship, Drug ; Egtazic Acid/pharmacology ; Hyaluronic Acid/pharmacology* ; Mice ; Mitogens/pharmacology* ; Phosphoproteins/metabolism* ; Phosphorylation ; Proto-Oncogene Proteins c-myc/metabolism* ; Retinoblastoma Protein/metabolism* ; Signal Transduction ; Sodium-Hydrogen Antiporter/antagonists & inhibitors ; Tyrosine

OBJECTIVETo observe the effect of an external film of hyaluronic acid (HA) on the rats wound healing.

METHODSForty-eight SD rats were randomly separated into eight groups of 6 rats each. Bilateral dorsal cuts were performed on each rat, left wound was used as the experiment with HA external film and right wound was used as the control only with normal saline. The process of healing was observed histologically following 1st, 3rd, 5th, 7th, 9th, 14th, 21st, and 28th days postoperatively.

RESULTSInflammation was lighter and epidermal healing was faster in the experimental group than those in the control. The fibroblasts degenerated and the collagen fiber changed to slim and loose bunches in the experimental group.

CONCLUSIONThe results indicated that HA external film could have powerful infiltrating activity at the early stage of wound healing, it could accelerate the healing of epidermis and delay the formation of keratinization layer.

Animals ; Collagen ; metabolism ; Drug Administration Routes ; Fibroblasts ; drug effects ; metabolism ; Hyaluronic Acid ; administration & dosage ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Treatment Outcome ; Wound Healing ; drug effects