Molecular Hyaluronic Acid Research Articles

Automated Open-Loop in Vitro photothermal TherapyUsing Near-Infrared Emitting Graphene Quantum Dots

Graphene-based materials have been the subject of interest for photothermal therapy due to their high light to heat conversion efficiency. Graphene quantum dots (GQDs) are expected to both possess advantageous photothermal properties and facilitate visible and near-infrared (NIR) fluorescence image-tracking, while surpassing the other graphene-based materials in their biocompatibility. To test these capabilities several different GQD structures were synthesized using top-down (RGQDs from reduced graphene oxide) and bottom-up (HGQDs from molecular hyaluronic acid) approaches. RGQDs and HGQDs possess nanometer-scale sizes, are hydrophilic, and exhibit fluorescence throughout the visible and NIR. The high biocompatibility of RGQDs (1.5 mg/ml) and HGQDs (1.7 mg/ml) and substantial absorption tail in the NIR make them suitable for biological photothermal therapy applications. In aqueous suspensions of RGQDs and HGQDs, a low power (0.8 W/cm2) 808 nm NIR laser irradiation facilitates heating up to 46.0 and 47.0 °C, respectively, making them sufficient for cancer tumor ablation. In vitro photothermal studies were performed via cost-effective open-loop automated approach involving a 3D printer head controlling the laser and a thermocouple. These experiments are carried out in a 96 well plate to show that GQDs can facilitate HeLa cancer cells heating up to 54.5 and 43.5 °C from 37.0 °C, leading to the inhibition of cell viability down to 23%. GQDs’ fluorescence in the visible and NIR effectively tracks their intracellular accumulation verifying effects of photothermal treatment. The combination of therapeutic and imaging capabilities makes GQDs developed in this work highly prospective anticancer agents.

Automated Approach to In Vitro Image-Guided Photothermal Therapy with Top-Down and Bottom-Up-Synthesized Graphene Quantum Dots.

Graphene-based materials have been the subject of interest for photothermal therapy due to their high light-to-heat conversion efficiency. Based on recent studies, graphene quantum dots (GQDs) are expected to possess advantageous photothermal properties and facilitate fluorescence image-tracking in the visible and near-infrared (NIR), while surpassing other graphene-based materials in their biocompatibility. Several GQD structures including reduced graphene quantum dots (RGQDs) derived from reduced graphene oxide via top-down oxidation and hyaluronic acid graphene quantum dots (HGQDs) hydrothermally bottom-up synthesized from molecular hyaluronic acid were employed to test these capabilities in the present work. These GQDs possess substantial NIR absorption and fluorescence throughout the visible and NIR beneficial for in vivo imaging while being biocompatible at up to 1.7 mg/mL concentrations. In aqueous suspensions, RGQDs and HGQDs irradiated with a low power (0.9 W/cm2) 808 nm NIR laser facilitate a temperature increase up to 47.0 °C, which is sufficient for cancer tumor ablation. In vitro photothermal experiments sampling multiple conditions directly in the 96-well plate were performed using an automated simultaneous irradiation/measurement system developed on the basis of a 3D printer. In this study, HGQDs and RGQDs facilitated the heating of HeLa cancer cells up to 54.5 °C, leading to the drastic inhibition of cell viability from over 80% down to 22.9%. GQD's fluorescence in the visible and NIR traces their successful internalization into HeLa cells maximized at 20 h suggesting both extracellular and intracellular photothermal treatment capabilities. The combination of the photothermal and imaging modalities tested in vitro makes the GQDs developed in this work prospective agents for cancer theragnostics.

Conservative treatment of hip osteoarthritis; comparison between three medium/high molecular weight hyaluronic acid injections and two injections of HYADD®4: a randomized controlled double-blind study.

Conservative treatment of hip osteoarthritis; comparison between three medium/high molecular weight hyaluronic acid injections and two injections of HYADD®4: a randomized controlled double-blind study.

Hyaluronic acid conjugated polydopamine functionalized mesoporous silica nanoparticles for synergistic targeted chemo-photothermal therapy.

The integration of chemotherapy and photothermal therapy into one nanoplatform has attracted much attention for synergistic tumor treatment, but the practical clinical applications were usually limited by their synergistic effects and low selectivity for disease sites. To overcome these limitations, a tumor-specific and pH/NIR dual-responsive multifunctional nanocarrier coated with mussel inspired polydopamine and further conjugated with targeting molecular hyaluronic acid (HA) was designed and fabricated for synergistic targeted chemo-photothermal therapy. The synthesized versatile nanoplatform displayed strong near-infrared absorption because of the successful formation of polydopamine coating. Furthermore, the nanosystem revealed high storage capacity for drugs and pH/NIR dual-responsive release performance, which could effectively enhance the chemo-photothermal therapy effect. With this smart design, in vitro experimental results confirmed that the drug loaded multifunctional nanoparticles could be efficiently taken up by cancer cells, and exhibited remarkable tumor cell killing efficiency and excellent photothermal properties. Meanwhile, significant tumor regression in the tumor-bearing mice model was also observed due to the combination of chemotherapy and photothermal therapy. Thus, this work indicated that the simple multifunctional nanoplatform can be applied as an efficient therapeutic agent for site-specific synergistic chemo-photothermal therapy.

Advances in the Production of High Molecular Weight Hyaluronic Acid by Microbial Fermentation

Advances in the Production of High Molecular Weight Hyaluronic Acid by Microbial Fermentation

Preparation and Characterization of Sponge Using Different Ratios of High Molecular Hyaluronic Acid and Small Intestine Submucosa

Preparation and Characterization of Sponge Using Different Ratios of High Molecular Hyaluronic Acid and Small Intestine Submucosa

Efficacy and Tolerance of an Injectable Medical Device Containing Hyaluronic Acid and Amino acids: A Monocentric Six-Month Open-Label Evaluation

Background: An innovative injectable solution containing low molecular hyaluronic acid (HA) and a specific Amino Acids mixture was formulated to physiologically promote local neo-collagenesis and elastogenesis through fibroblasts chemotaxis migration into the injected area. Objective: Aim of this open clinical trial is to evaluate efficacy and tolerance of the injectable product under study, on main sign of face skin photoaging. Methods: A single Italian center treated 25 female subjects aged 48-65 years with 4 micro-injection sessions with 10-day time laps between each product administration. Subjects were evaluated in basal conditions and after 4, 8, 12 and 24 weeks, using validated clinical scales, subjective evaluations and objective quantitative outcome measures. Assessment of aesthetic results included photographic documentation. Results: Obtained results showed an improvement of all the clinical and subjective assessments and on the majority of objective instrumental parameters. These were already significant 10 days after the first injection procedure and were still significant and still improving after 6 months (at follow up). Global judgment on tolerability was good/excellent, both in the investigators’ opinion and volunteers’ self-evaluation. Conclusions: Obtained results confirm the aesthetic performance of the injectable product on main signs of face skin photoaging. In particular, it was demonstrated the biovolumetric effect, the antiwrinkle efficacy, a superficial and deep moisturising activity and elasticizing properties. This study supports the definition of extracellular matrix targeting (ECM-targeting) for this product.

Global facial enhancement using a cross-linked hyaluronic acid dermal filler

Molecular hyaluronic acid (HA) has been a staple product in aesthetics for many years. The Decoria range of dermal fillers, manufactured by Bohus BioTech, represents an innovation in HA dermal filler manufacturing using unique Decoria Proprietary Spherification (DPS) technology. Spherification of the HA particles results in smoother results and higher rates of patient satisfaction. There are two products in the Decoria range (Decoria Essence and Decoria Intense), covering a range of facial treatment indications. This article introduces the technology of the products and describes the way in which Decoria dermal fillers can be used for global facial rejuvenation. Three case studies were undertaken by the author, addressing a diverse range of patients' cosmetic concerns, including lip enhancement, mid-face revolumisation and lower facial contouring. The outcomes of these case studies, illustrated with before and after photography, demonstrate excellent aesthetic results and high patient satisfaction.

Hydroxyapatite combined with hyaluronic acid metronidazole gel increased the quantity of osteoblasts in the alveolar bone wistar rat

Background: Bone graft material have been used extensively in bone healing and periodontal treatment. Alloplast such as hydroxyapatite are frequently used to repair and reconstruct bone defects. By merely applying hydroxyapatite for the treatment of bone is not fully effective yet to produce new bone regeneration. Locally applied high molecular hyaluronic acid (HA) has been shown to stimulate differentiation and migration of mesenchymal cells. Recent studies on regenerative surgical procedures indicate that reduction of bacterial burden at the wound site may improve the clinical outcome of regenerative therapy. Metronidazole has the greatest bacteriostatic effect. A clinical application of HA metronidazole gels during the surgical therapy may reduce the bacterial contamination of surgical wound site. Purpose: The purpose of this study was to examine the effect of combination hydroxyapatite and HA metronidazole gel 1% on osteoblast cell number after wound healing process in the wistar rats incisor tooth extraction socket. Method: Twenty seven wistar rats were divided randomly into 3 groups. The first group consisted of wistar rats given hydroxyapatite were subjected to the mandibular incisor extraction socket. The second group were given hydroxyapatite combined with HA metronidazole gel 1%. The control group were filled with blood. Wistar rats were euthanized on day 14 and then preparation for histological examination was stained using hematoxylin-eosin and then the numbers of the osteoblasts were calculated. Result: The differences in each group were tested by one way Anova test ( α =0.05). The numbers of osteoblasts in each group had a significant difference (p<0.05): the highest numbers of osteoblasts were found in the group that was given hydroxyapatite combined with HA metronidazole, followed by the lower numbers of osteoblasts in the hydroxyapa ite group and the lowest numbers of osteoblasts were found in the control group. Conclusion: Adjunctive application of HA metronidazole to hydroxyapatite after tooth extraction increase the number of osteoblast in the treatment of bone defects on a wistar rat model.

AB0066 Cyclic phosphatidic acid (cpa) stimulates hyaluronic acid production in human osteoarthritic articular chondrocytes, and intra-articular administration of cpa suppresses pain, swelling, and cartilage destruction in rabbit experimental osteoarthritis.

BackgroundCyclic phosphatidic acid (cPA) is one of lipid mediators, has been shown various biological effects1. On human skin fibroblasts, cPA stimulates high molecular hyaluronic acid (HA) production through up-regulating HA...

Reflective account of a delayed adverse reaction to a large molecular filler

This is a reflective account of a 62-year-old woman (Jean) who developed delayed hypersensitivity reactions to a large molecular hyaluronic acid (HA) filler in 2009. Jean had previous injections in August 2008 of 1 ml of HA filler to her nasolabial folds, of 1 ml of HA filler to the peri-oral area in November 2008 and 1 ml of HA filler to nasolabial folds in May 2009, all with excellent results and high patient satisfaction. However, Jean presented 10 weeks after her initial treatment for mid-face volume loss with a volumising HA dermal filler and developed an adverse delayed reaction. Following this experience, the author formed a protocol for her clinic and has taken part in discussions about protocols in practice, prescribing and managing complications.

BMP-2 and BMP-7: differential regulation of chondrogenic differentiation

BMP-2 and BMP-7: differential regulation of chondrogenic differentiation

Establishment of novel three-dimensional culture system for human chondrocytes using micro-space plate

Purpose: It has been reported that monolayer culture of human articular chondrocytes results in dedifferentiation and loss of chondrogenic property. Therefore, the dedifferentiation is a big problem when the influence of cytokines and drugs were evaluated. In order to solve this problem, we performed a novel three-dimensional culture system for human chondrocytes using micro-space plates compared with monolayer culture. Methods: The micro-space plate has regularly arranged 200 mm×100 mm×50 mm square compartments on its surface. Human articular chondrocytes were cultured in flat plates or micro-space plates for 1 week. After culture, the level of collagen 1A1 (COL1A1) and aggrecan mRNA expressions were measured by real-time PCR, and cartilage matrix production was assessed by alcian blue staining. Furthermore, chondrocytes were cultured with IL-1β, IL-6+soluble IL-6 receptor (sIL-6R) in the presence or absence of high molecular hyaluronic acid (HA) for additional 24 h and then the production of MMP-3 in cell supernatant was measured by ELISA. Results: At the same passage of cells, human articular chondrocytes grown on micro-space plates in clusters after 5-7 days’ culture, whereas cells grown on flat plates in fibroblast-like morphology. In three-dimensional culture, the mRNA expression levels of COL1A1 and aggrecan were higher than those in monolayer culture. And the cartilage matrix production in three-dimensional culture was higher than that in monolayer culture. Moreover, the production of MMP-3 induced by IL-1β and IL-6+sIL-6R was higher in three-dimensional culture than in monolayer culture. Inhibitory activity of HA on MMP-3 production was similar between three-dimensional culture and monolayer culture. Over the subsequent passage, the expression levels of COL1A1 and aggrecan mRNA and MMP-3 production by IL-1β and IL-6+sIL-6R gradually decreased in monolayer culture, but they remained at higher levels in three-dimensional culture. Conclusions: In conclusion, the present study demonstrated that the novel three-dimensional culture using micro-space plates maintains chondrogenic properties. Three-dimensional culture using micro-space plates is a useful tool for chondrocyte culture.

The effect of high molecular hyaluronic acid on the induction of matrix degradation enzymes By IL-6, IL-1β and TNF-α

The effect of high molecular hyaluronic acid on the induction of matrix degradation enzymes By IL-6, IL-1β and TNF-α

Wound healing potential by hyaluronate gel in streptozotocin-induced diabetic rats

This study was conducted to investigate whether topical application of Hyaluronate gels could improve the impaired wound healing in streptozotocin-induced diabetic rats. Four groups of adult male Sprague Dawley rats, 2 cm full-thickness skin wound were experimentally created on the posterior neck area of streptozotocin-induced diabetic rats (STZ). Wounds of Group 1 animals were topically treated with the vehicle, gum acacia in normal saline, as a placebo control group. Group 2 animals served as reference standard and treated topically with Intrasite gel. Animals of Group 3, 4 were treated topically with new oral high molecular weight hyaluronic acid 240 mg/100 g gel, 0.8% hyaluronic acid gel respectively. Macroscopically, the reference standard gel and the three gels-treated wounds were significantly healed faster in comparison to placebo control wounds group. Wound closure was significantly accelerated by topical application of high molecular weight hyaluronic acid compared to reference standard gel, 0.8% hyaluronic acid gel. Furthermore, histological examination of healed wounds with reference standard gel, 0.8% hyaluronic acid gel, and hyaluronic acid 240 mg/100 g gel-treated wounds revealed comparatively increases in macrophages, fibroblast migration, collagen regeneration, and epithlization compared with the placebo control group. In conclusion, the new oral high molecular weight hyaluronic acid gel can improve the impaired healing of diabetic wounds and could be useful in treating oral ulcerations.

Specific degradation of hyaluronic acid and its rheological properties

High molecular hyaluronic acid was degraded by ultrasonication to definite molecular weight varying from 0.4 to 2.0 × 10 6g mol −1. The weight average molecular weight (M w) of degraded hyaluronic acid in 0.2 M NaCl was determined by size exclusion chromatography combined with multi-angle-laser-light-scattering (SEC-MALLS). Comparison measurements by using an Ubbelohde viscosimeter and a low shear rate viscosimeter (Zimm-Crothers) were performed for all sonicated samples. The relation of M w of the hyaluronic acid to the intrinsic viscosity [η] was calculated according to the Mark-Houwink equation, [η] = K M w α. The coefficients K = 5.075 10 −2 and α = 0.716 determined by Ubbelohde and K = 3.023 10 −2 and α = 0.770 by Zimm-Crothers viscosimeter. Furthermore, the dependence on pH and the influence of bivalent cations were investigated. In the range of pH 5.0 to 11.0 the intrinsic viscosity [η] was constant. At pH less than 5.0, the intrinsic viscosity [η] and the molecule volume decreased probably due to the protonation of glucoronic acid groups. Most bivalent cations do not affect the rheological behaviour significantly, only Fe 2+ causes to decrease of the intrinsic viscosity [η] and the molecule volume by intramolecular chain linkage combined with displacement of hydrating water.

The glycosaminoglycans in cultures of stimulated rat peritoneal macrophages. 2. Gel chromatographic studies and the behaviour of heparan sulfate.

The molecular weight distribution of pMP-derived glycosaminoglycans (GAG), i.e. non-sulfated GAG, chondroitin sulfate (CS), and heparin sulfate (HS)-like material was determined. The peritoneal macrophages (pMP) were harvested from rats normal or stimulated by i.p. injection of thioglycolate, carrageenan or BCG, and maintained in culture. The GAG of cell layer and medium were isolated separately after labeling with 35S-sulfate and 3H-acetate. Treatment with nitrous acid served to remove HS-like material. Labeling with 3H-acetate served to detect synthesis of the high m. w. hyaluronic acid (HA). Gel chromatic separation was done using Sephadex G-200 columns. The maximal size of 35S-labeled GAG, especially HS (36 kDa), was reduced in cultural medium and cell layer after stimulation in vivo. Reduction was most pronounced after application of carrageenan followed by thioglycolate and BCG/LPS stimulation. The extracellular GAG of BCG-stimulated pMP were smallest, probably due to degradation. Heparan sulfate-like material made up a larger proportion in monolayer and medium, comprising the total m.w. range up to 36 kDa. The GAG sensitive to nitrous acid were maximal in cultures of carrageenan-stimulated pMP and minimal in those of thioglycolate-stimulated pMP. This type of HS was sensitive to hyaluronidase, too. Any synthesis of high molecular hyaluronic acid was not found in normal or stimulated rat pMP. Therefore MP-associated HA must be adsorbed from other sources or synthesized by early forms of macrophages.