[en] Highlights: • This study introduced an in situ injectable hyaluronic acid/gelatin hydrogel, which was prepared by chemically crosslinking hyaluronic acid and gelatin with EDC/NHS. • The hydrogel had the short gelatin time, good cytocompatibility, strong bursting strength and well hemostatic ability. • The hemostatic ability of HA/G hydrogel was same as that of fibrin glue. It suggested that the hydrogel could be a tissue sealant for hemorrhage control in clinic. -- Abstract: Tissue sealants are used for hemorrhage control which is imperative in many surgical procedures. It is a highly challenging task to obtain the ideal tissue sealant. Only a few commercially tissue sealants are available to be used for internal tissue or organ hemorrhage control. This study introduced two in situ injectable hydrogels for hemorrhage control: self-crosslinking gelatin (sc-G) hydrogel and hyaluronic acid/gelatin (HA/G) hydrogel. They were prepared on the tissue surface in situ and characterized by rheological analysis, stability, cytotoxicity, and bursting strength test. The hemostatic ability of the hydrogels was evaluated in a liver-bleeding rat model. The sc-G and HA/G hydrogels gelled around 90 s and 50 s, respectively. They were preferable for cell attachment and proliferation. The bursting strengths of both hydrogels exceeded that of fibrin glue. The hemostatic ability of HA/G hydrogel was better than that of sc-G hydrogel, and was same as that of fibrin glue. The HA/G hydrogel could be used as a tissue sealant for hemorrhage control in clinic.

[en] Highlights: • Microfibrous scaffolds were electrospun from a blend of collagen, hyaluronic acid and poly(l-lactide-co-ε-caprolactone) • The PLC/COL/HA scaffolds had smaller pore size and higher water uptake capacity than PLC scaffolds • The PLC/COL/HA scaffolds had a Young’s Modulus similar to that of many soft tissues • The PLC/COL/HA scaffolds better supported cell adhesion when compared with PLC scaffolds • The PLC/COL/HA scaffolds induced formation of interconnected capillaries -- Abstract: Success of 3D tissue substitutes in clinical applications depends on the presence of vascular networks in their structure. Accordingly, research in tissue engineering is focused on the stimulation of angiogenesis or generation of a vascular network in the scaffolds prior to implantation. A novel, xeno-free, collagen/hyaluronic acid-based poly(l-lactide-co-ε-caprolactone) (PLC/COL/HA) (20/9.5/0.5 w/w/w) microfibrous scaffold was produced by electrospinning. Collagen types I and III, and hyaluronic acid were isolated from human umbilical cords and blended with the GMP grade PLC. When compared with PLC scaffolds the PLC/COL/HA had higher water uptake capacity (103% vs 66%) which may have contributed to the decrease in its Young's Modulus (from 1.31 to 0.89 MPa). The PLC/COL/HA better supported adipose tissue-derived mesenchymal stem cell (AT MSC) adhesion; within 24 h the cell number on the PLC/COL/HA scaffolds was 3 fold higher. Co-culture of human umbilical vein endothelial cells and AT MSCs induced capillary formation on both scaffold types, but the PLC/COL/HA led to formation of interconnected vessels whose total length was 1.6 fold of the total vessel length on PLC. Clinical use of this scaffold would eliminate the immune response triggered by xenogeneic collagen and transmission of animal-borne diseases while promoting a better vascular network formation.

[en] Highlights: • Assessment of the subcutaneous degradation process of insoluble hyaluronic acid. • The speed of degradation of in vivo insoluble hyaluronic acid membranes varies depending on their swelling rate. • The process of degradation of insoluble hyaluronic acid in the subcutaneous over time was elucidated. Insoluble hyaluronic acid (IHA) may prevent adhesions by forming a physical barrier during the period when postoperative adhesions form. This study was performed to verify the changes that a solid IHA membrane undergoes as it is degraded in vivo, and to ascertain the swelling rate of IHA required for it to function as a physical barrier during the postoperative adhesion formation period. Nine female WI rats weighing 300–400 g were used. Discs 8 mm in diameter were cut out of dry IHA membranes made of IHA with a swelling rate (wet weight/dry weight) of either 2.47 (high-swelling IHA) or 1.94 (low-swelling IHA). They were placed in saline to swell and then washed with saline before subcutaneous implantation in four pockets in each rat. The high-swelling IHA started to degrade more rapidly than the low-swelling IHA. There was no evidence of degradation of the low-swelling IHA until day 7, but once it had started, the speed of degradation tended to be similar to that of the high-swelling IHA. The present results showed that, when IHA is implanted subcutaneously in rats, it is degraded over time in a phased process. The swelling rate required for the use of IHA as a postoperative adhesion barrier was also suggested.

[en] The configuration of charged polymers is heavily dependent on interactions with surrounding salt ions, typically manifesting as a sensitivity to the bulk ionic strength. In this work, we use single-molecule mechanical measurements to show that a charged polysaccharide, hyaluronic acid, reflects an interesting regime of insensitivity to ionic strength in the presence of trivalent ions. Using simulations and theory, we propose that this is caused by the formation of a “jacket” of ions, tightly associated with the polymer, whose charge (and thus effect on configuration) is robust against changes in solution composition.

[en] Hyaluronic acid (HA), a principal matrix molecule in many tissues, is present in high amounts in articular cartilage. HA contributes in unique ways to the physical behavior of the tissue, and has been shown to have beneficial effects on chondrocyte activity. The goal of this study was to incorporate graduated amounts of HA into type I collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis in vitro. The results demonstrated that the amount of contraction of HA/collagen scaffolds by adult canine articular chondrocytes increased with the HA content of the scaffolds. The greatest amount of chondrogenesis after two weeks was found in the scaffolds which had undergone the most contraction. HA can play a useful role in adjusting the mechanical behavior of tissue engineering scaffolds and chondrogenesis in chondrocyte-seeded scaffolds

No abstract available

[en] Highlights: • A new approach for the synthesis of catechol-conjugated hyaluronic acid (HA-Cat) macromers with high substitution degree. • HA-Cat hydrogels show excellent adhesion to biomacromolecules, cells, and tissues. • The BMP-2-immobilized HA-Cat hydrogels can effectively promote osteogenesis of seeded hMSCs. -- Abstract: Hydrogels are promising soft materials for the delivery of therapeutic cells and cargo molecules. Inspired by mussel adhesion chemistry, hydrogels based on catechol (Cat)-modified polysaccharides have been developed to enhance hydrogel–tissue interactions. However, due to the inevitable side reactions such as self-polymerization of dopamine involved in the conventional catechol conjugation process, the efficiency of catechol conjugation to polymers is typically low, leading to insufficient stability, low mechanical strength, and poor adhesiveness of these catechol-modified hydrogels. In this study, we report a new approach to synthesize catechol-functionalized hyaluronic acid with improved degree of substitution. Due to the significantly increased conjugated Cat groups, the obtained HA-Cat hydrogels not only can adhere to tissue samples under wet conditions but also can capture cell adhesion proteins to enhance cell attachment and spreading. Meanwhile, owing to the extensive Cat–protein interactions, these hydrogels can also facilitate long-term release of protein-based therapeutic cargoes, such as the osteoinductive BMP-2 protein, thereby effectively promoting osteogenic differentiation of stem cells. These findings show that the HA-Cat hydrogels are ideal carriers of therapeutic cells and drugs for tissue regeneration.

[en] CD44 describes a family of surface proteins consisting of many isoforms due to alternative splice of ten 'variant' exons. Members of this family are involved in various processes including hematopoiesis, lymphocyte activation and homing, limb development, wound healing and tumor progression. Clinically, CD44 has been shown to be a prognostic factor for several human cancers. To answer the question which isoform might be relevant for tumor progression and to gain an insight into the mechanism of its function, I established transfectants of the LB lymphoma cell line in which the expression of four CD44 isoforms, namely CD44v3-10, CD44v4-10, CD44v8-10 and CD44s, was controlled by the Tet-off promoter. In the presence of Doxycycline, the expression was repressed. Removal of Doxycycline switched on expression and the maximal CD44 amount was obtained within two days. The transfectants were characterized regarding their ability to bind to the extracellular matrix component hyaluronate (HA). Overexpression of all four CD44 isoforms conferred the ability to bind HA on LB cells. Other glycosaminoglycans (GAGs) were bound in an isotype-specific fashion. CD44v3-10, CD44v4-10 and CD44v8-10 showed high binding affinity to chondroitin A, B and C, and low affinity to heparin, heparan sulfate and keratan sulfate. CD44s could not bind to these GAGs. Among these three variants, the binding ability of CD44v3-10 was the strongest. CD44 clustering seemed to play a crucial role for HA binding. Both CD44s and CD44v8-10 formed reduction-sensitive complexes in LB cells. The complexes are homooligomers or heterooligomers composed of different isoforms. Cys286 in CD44 transmember domain was not responsible for the formation of reduction-sensitive oligomer or for the enhanced HA binding in LB cell line. Using a conditional dimerization system the requirement of CD44 oligomerization for HA binding was directly demonstrated. The induction of oligomerization increased HA binding. Finally, I studied the role of CD44 in lymphoma development by subcutaneous injection of the transfectants in syngeneic or immunocompromised mice. I found no influence of CD44v3-10, CD44v4-10 and CD44v8-10 isoforms on the LB lymphoma formation and metastasis. (orig.)

[en] PVA was dissolved in mixed solvent (DMSO and water) and followed by several freeze-thaw cycles in a mold to produce PVA membrane. Surface modification of PVA membranes by HA molecules was investigated to improve the hydrophilicity of the membrane surface thereby reducing adsorption of the proteins onto the membrane. The surface composition, water contact angle, optical and mechanical properties, surface morphology, cell compatibility and protein adhesion were systematically investigated. ATR-FTIR spectra, XPS, SEM and AFM indicated that PVA membranes were successfully modified by grafting of the HA. The modified membranes showed increased hydrophilicity and cytocompatibility, decreased surface roughness and mechanical properties, and suppressed cell and protein adhesion compared to the pristine membrane. In general, the achievement of the HA coating with anti-adhesive property can potentially be widely used on surface modification of artificial cornea and other biomedical implants. (paper)

[en] The grafted human glioblastoma cell CB109 was used as a model for intralesional therapy with 131I-labelled hyaluronectin glycoprotein (131I-HN). 131I-HN bound specifically to in situ hyaluronic acid (HA), a main component of the extracellular matrix which is involved in tumour invasion. Labelling experimental conditions were determined and, finally, 25 μCi/μgHN, 1 μg chloramine-T/μgHN and a 60-s stirring period provided a 131I-HN preparation with an optimal affinity for HA (64% compared to unlabelled HN). Following intratumoral injection, 131I-HN was retained with a limited diffusion outside the tumour. On day 4 the radioactivity concentrated in the tumour was still 25 times greater than that in the liver, spleen and kidneys combined. For therapeutic assays, 65 μCi 131I-HN was injected into the tumour, resulting in a delivery of 6.8 Gy over a 7-day period. Controls received unlabelled HN, heat-inactivated HN, a mixture of inactivated HN plus free 131I or no treatment (six animals per group). Tumour volumes were evaluated every second day from treatment day and the rate of tumour growth was expressed as a ratio of tumour size at time intervals to the tumour size at the time of injection. Growth curves were compared: heat-inactivated with or without free 131I had no anti-tumour effect. Unlabelled HN-injected tumours had a slightly slower growth rate than untreated tumours (p < 0.02) and growth rate of 131I-HN-injected tumours was much lower (p < 0.00002). A pronounced inhibitory effect with intralesional 131I-labelled HN injection resulted from a combination of a) blockage of HA, a proliferation facilitating factor, and b) local irradiation of tumoral tissue, while uptake in normal tissues was minimized