OBJECTIVETo investigate the possible influence of the impairment of lymph fluid on the metabolism of hyaluronic acid (HA) in the lymphedematous skin tissue.

METHODSTissue fluid was collected in lymphedematous limbs and the contralateral healthy limbs of 39 patients and HA content was measured with radioimmunoassay. The protein contents were also measured.

RESULTSThe HA contents in interstitial fluid of lymphedematous limb were significantly (8 fold) higher than that of normal limb. The protein concentration in the tissue fluid did not show significant differences between lymphedema and those with normal tissue.

CONCLUSIONThe result suggests blockage of regional draining lymphatics may impairs breakdown of HA and the stagnation of HA in the limb may exert a deleterious effect on the interstitium.

Extracellular Fluid ; metabolism ; Forearm ; Humans ; Hyaluronic Acid ; metabolism ; Leg ; Lymphedema ; metabolism ; Radioimmunoassay ; Skin ; metabolism ; pathology

A hyaluronic acid (HA) incorporated porous collagen matrix was fabricated at -70 degree C by lyophilization. The HA incorporated collagen matrix showed increased pore size in comparison with collagen matrix. Biodegradability and mechanical properties of matrices were controllable by varying the ultraviolet (UV) irradiation time for cross-linking collagen molecules. Addition of HA to collagen matrix did not effect ultimate tensile stress after UV irradiation. HA incorporated collagen matrices demonstrated a higher resistance against the collagenase degradation than collagen matrix. In an in vitro investigation of cellular behavior using dermal fibroblasts on the porous matrix, HA incorporated collagen matrix induced increased dermal fibroblast migration and proliferation in comparison with collagen matrix. These results suggest that the HA incorporated collagen porous matrix assumes to enhance dermal fibroblast adaptation and regenerative potential.
Collagen/*metabolism ; Extracellular Matrix/*metabolism ; Fibroblasts/*physiology ; Human ; Hyaluronic Acid/*metabolism ; Porosity

The theory of channels and collaterals is a very important "substance" prop for the basic theories of TCM. After component model of substances of channels and collaterals is raised, according to the principle, structure and function are dialectical unity, the author finds in deeply research on a great number of completed experimental investigations of corresponding anatomic position of channels and collaterals and similar physiology at home and abroad that the difficult to understand TCM concepts such as "interdependence between yin and yang", "defensive energy" all could be displayed by the substance of body liquid showing liquid crystalline state during living of channels and collaterals.
Cell Membrane ; metabolism ; Cells ; metabolism ; Humans ; Hyaluronic Acid ; metabolism ; Medicine, Chinese Traditional ; Meridians ; Models, Biological

OBJECTIVETo analyze the quality characteristics of human spermatozoa with hyaluronic acid (HA) receptors and search for a new indicator for the assessment of sperm quality.

METHODSUsing sperm-HA binding assay with HA-coated slides, we determined the binding rate of motile sperm with HA receptors and analyzed its correlation with routine semen parameters, sperm membrane function, sperm fertilizing function and diminished/arrested sperm maturation.

RESULTSThe motile sperm with HA binding sites in the acrosomal region showed significantly higher acrosomal integrity ([95.4 +/- 3.9]%) and mitochondrial membrane potential (MMP) ([97.8 +/- 2.1]%) than those in the initial semen ([68.8 +/- 6.2]% and [72.8 +/- 7.4]%) (P < 0.01). The sperm-HA binding scores were correlated mildly with many routine semen parameters (r = 0.195-0.268, P < 0.05), positively with the acrosome reaction level after ionophore challenge (r = 0.666, P < 0.01) and normal sperm morphology (r = 0.417, P < 0.01), and negatively with sperm nucleoprotein immaturation (r = -0.266, P < 0.01), DNA fragmentation (r = -0. 308, P < 0.01) and excessive residual cytoplasm (r = -0.218, P < 0.05).

CONCLUSIONSperm with HA receptors in the acrosomal region exhibit significant advantages in plasma membrane structure, fertilizing potential and maturation. The sperm-HA binding assay, which is based on a relationship between sperm receptors for zona pellucida and HA, is likely to become a new independent indicator for assessing the multiple qualities of spermatozoa.

Acrosome ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Hyaluronic Acid ; Male ; Spermatozoa ; cytology ; metabolism ; physiology

Hyaluronic acid (HA) is widely used in many fields, such as medicine, cosmetics and food. The bioactivity of HA depends on its molecular weight (Mw). Owing to the important physiological activities and special physiological functions, HA oligosaccharides have important application prospects in medicine fields. Streptococcus zooepidemicus has wide applications in commercial production of HA, due to its short fermentation cycle and strong production intensity. In order to efficiently synthesize HA oligosaccharides and solve the dissolved oxygen in the fermentation process, in this study, we overexpressed HA synthase (HasA) and introduced and optimized the leech hyaluronidase LHAase in Streptococcus zooepidemicus WSH-24. As a result, HA oligosaccharides were efficiently produced with improved dissolved oxygen. After 24 h, HA oligosaccharides production intensity reached to 294.2 mg/(L·h), and the concentration accumulated to 0.97 g/L in flask cultures, which was 1.82 times of the wild strain. Impressively, HA oligosaccharides were increased to 7.06 g/L in 3 L fermentor. The constructed Streptococcus zooepidemicus strain for producing HA oligosaccharides would have broad application prospects.
Bioreactors ; Fermentation ; Hyaluronan Synthases ; genetics ; metabolism ; Hyaluronic Acid ; genetics ; metabolism ; Industrial Microbiology ; Oligosaccharides ; genetics ; metabolism ; Streptococcus equi ; genetics ; metabolism

OBJECTIVETo investigate the distribution of hyaluronic acid (HA) with iohexol tracing in the knee joint cavity of rabbits using CT plain scan, three-dimensional reconstruction and Χ-ray and observe how different injection sites affect HA distribution.

METHODSMixtures of HA and iohexol (tracer) were prepared that contained final iohexol concentrations of 2.5%, 5%, 10%, 20%, or 40%. The HA-iohexol mixtures (0.5 ml) were injected into rabbit knee joints, and the optimal iohexol concentration that allowed clear differentiation of the injected agents from the surrounding tissues was determined using dual-source CT plain scan and three-dimensional reconstruction technique. The HA-iohexol mixture (0.5 ml) containing the optimal concentration of iohexol was then injected into the knees of the rabbits either through the patella medial approach or the medial joint line approach, and HA distribution in the knee joint cavity was observed using CT scan and Χ-ray.

RESULTSThe CT value of HA-iohexol mixture increased progressively with the tracer concentration. After injection of the mixture containing 2.5%, 5%, 10%, 20%, and 40% iohexol, the CT value ratios of the soft tissue, HA-iohexol mixture and bone cortex were 2:7:46, 2:14:44, 2:28:44, 2:60:46, and 2:98:45, respectively, and a iohexol concentration of 5% was determined as optimal for differntiating the injected agents from the surrounding tissues. The HA-iohexol mixutre containing 5% iohexol injected through the medial-patellar approach was distributed mainly over the patello-femoral joint, and that injected through the joint line approach was found mainly over the tibio-femoral joint.

CONCLUSIONHA-iohexol mixture containing 5% iohexol allows clear differentiation of bone cortex and soft tissues in rabit knee joint from the injected agents on CT scan and Χ-ray, and the injection approach can influence HA distribution in the knee joint cavity.

Animals ; Contrast Media ; Hyaluronic Acid ; metabolism ; Iohexol ; Knee Joint ; Rabbits ; Tissue Distribution ; Tomography, X-Ray Computed

OBJECTIVE: To study the expression of hyaluronan synthasel-3 (HAS1-3) in nasal polyps, and discuss their clinical significance. METHOD: The expression of HA, HAS1, HAS2 and HAS3 in nasal polyps group (25 cases) and inferior turbinate group (15 cases) was detected by immunohistochemistry and semi-quantitative RT-PCR. RESULT: Compared with the control, the expression of HA, HAS1 and HAS3 was higher in nasal polyps (P<0. 05), while there was no significant difference in HAS2 expression (P>0. 05). CONCLUSION The increase of HASI and HAS3 expression may be the main cause of the excessive deposition of HA in nasal polyps, which may play an important role in the pathogenic process of nasal polyps, and become a potential target for therapy of nasal polyps.
Glucuronosyltransferase ; metabolism ; Humans ; Hyaluronan Synthases ; Hyaluronic Acid ; Immunohistochemistry ; Nasal Polyps ; enzymology

The high- and the low-molecular weight hyaluronic acids (HMW-HA and LMW-HA, respectively) showed different biological activities in inflammation. However, the role of LMW-HA in inflammatory response is controversial. In this study, we aimed to investigate the effect of bioactive hyaluronan (B-HA) on lipopolysaccharide (LPS)-induced inflammatory responses in human macrophages and mice. B-HA was produced from HA treated with glycosylated recombinant human hyaluronidase PH20. Human THP-1 cells were induced to differentiate into macrophages. THP-1-derived macrophages were treated with B-HA, LPS, or B-HA + LPS. The mRNA expression and the production of inflammatory cytokines were determined using quantitative real-time PCR and enzyme-linked immunosorbent assay. The phosphorylation levels of proteins in the nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), and IRF-3 signaling pathways were measured using Western blot. The in vivo efficacy of B-HA was assessed in a mouse model of LPS-induced inflammation. Results showed that B-HA inhibited the expression of TNF-α, IL-6, IL-1, and IFN-β, and enhanced the expression of the antiinflammatory cytokine IL-10 in LPS-induced inflammatory responses in THP-1-derived macrophages and in vivo. B-HA significantly suppressed the phosphorylation of the TLR4 signaling pathway proteins p65, IKKα/β, IκBα, JNK1/2, ERK1/2, p38, and IRF-3. In conclusion, our results demonstrated that the B-HA attenuated the LPS-stimulated inflammatory response by inhibiting the activation of the TLR4 signaling pathway. B-HA could be a potential anti-inflammatory drug in the treatment of inflammatory disease.
Animals ; Cytokines ; Hyaluronic Acid ; Lipopolysaccharides ; Mice ; NF-kappa B/metabolism* ; Signal Transduction ; Toll-Like Receptor 4

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

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