[The in vivo thrombosis evaluation for the biodegradable polymer stent].

New biodegradable intravascular stent can reduce risk of foreign bodies retained, thus, it is widely concerned and some of the products have been introduced into the clinic. However, the characteristic of biodegradable may lead to more safety concerns associated with thrombosis. To ensure the safety, the thrombus formation experiment in vivo needs to be carefully designed and evaluated based on GB/T 16886.4 standard, but current standard do not provide explicit testing and evaluating methods. Establishing animal model with experimental pigs, the study compares biodegradable coronary stents and metal stents by simulating clinical implantation in vivo on the thrombus formation in the implanting process, and after the short-term and long-term implantation. The evaluation methods include gross observation, digital subtraction angiography intraoperative analysis, optical coherence tomography analysis, scanning electron microscopy and so on. The results show that combining these methods could comprehensively evaluate the whole process of the thrombus formation from the beginning of implantation to the end of preclinical animal experiments, so that, it may better predict the clinical thrombosis risk, and the selection of the control was very important. The study tries to use the comparison examples of thrombosis on the new medical instrument to provide the clue for thrombosis evaluation in vivo on similar instruments and show the methodology on the preclinical evaluation.

A New Absorbable Synthetic Substitute With Biomimetic Design for Dural Tissue Repair.

Dural repair products are evolving from animal tissue-derived materials to synthetic materials as well as from inert to absorbable features; most of them lack functional and structural characteristics compared with the natural dura mater. In the present study, we evaluated the properties and tissue repair performance of a new dural repair product with biomimetic design. The biomimetic patch exhibits unique three-dimensional nonwoven microfiber structure with good mechanical strength and biocompatibility. The animal study showed that the biomimetic patch and commercially synthetic material group presented new subdural regeneration at 90 days, with low level inflammatory response and minimal to no adhesion formation detected at each stage. In the biological material group, no new subdural regeneration was observed and severe adhesion between the implant and the cortex occurred at each stage. In clinical case study, there was no cerebrospinal fluid leakage, and all the postoperation observations were normal. The biomimetic structure and proper rate of degradation of the new absorbable dura substitute can guide the meaningful reconstruction of the dura mater, which may provide a novel approach for dural defect repair.

Influences of the steam sterilization on the properties of calcium phosphate porous bioceramics.

The influences of steam sterilization on the physicochemical properties of calcium phosphate (Ca-P) porous bioceramics, including ß-tricalcium phosphate (ß-TCP), biphasic calcium phosphate (BCP) and hydroxyapatite (HA) are investigated. After being steam sterilized in an autoclave (121 °C for 40 min), the porous bioceramics are dried and characterized. The steam sterilization has no obvious effects on the phase composition, thermal stability, pH value and dissolubility of ß-TCP porous bioceramic, but changes its morphology and mechanical strength. Meanwhile, the steam sterilization leads to the significant changes of the morphology, phase composition, pH value and dissolubility of BCP porous bioceramic. The increase of dissolubility and mechanical strength, the decrease of pH value of the immersed solution and partial oriented growth of crystals are also observed in HA porous bioceramic after steam sterilization. These results indicate that the steam sterilization can result in different influences on the physicochemical properties of ß-TCP, BCP and HA porous bioceramics, thus the application of the steam sterilization on the three kinds of Ca-P porous bioceramics should be considered carefully based on the above changed properties.

Polyvinyl-alcohol, chitosan and graphene-oxide composed conductive hydrogel for electrically controlled fluorescein sodium transdermal release.

Accurate and controlled release of drug molecules is crucial for transdermal drug delivery. Electricity, as an adjustable parameter, offers the potential for precise and controllable drug delivery. However, challenges exist in selecting the appropriate drug carrier, electrical parameters, and release model to achieve controlled electronic drug release. To overcome these challenges, this study designed a functional hydrogel using polyvinyl alcohol, chitosan, and graphene oxide as components that can conduct electricity, and constructed a drug transdermal release model using fluorescein sodium salt with proper electrical parameters. The results demonstrated that the hydrogel system exhibited low cytotoxicity, good conductivity, and desirable drug delivery characteristics. The study also integrated the effects of drug release and tissue repair promotion under electrical stimulation. Cell growth was enhanced under low voltage direct current pulses, promoting cell migration and the release of VEGF and FGF. Furthermore, the permeability of fluorescein sodium salt in the hydrogel increased with direct current stimulation. These findings suggest that the carbohydrate polymers hydrogel could serve as a drug carrier for controlled release, and electrical stimulation offers new possibilities for functional drug delivery and transdermal therapy.

A chitosan-optimized polyethyleneimine/polyacrylic acid multifunctional hydrogel for reducing the risk of ulcerative arterial bleeding.

Ulcerative arterial bleeding is characterized by sudden onset, rapid disease development, and high mortality, which is a great challenge for clinicians to treat, specially bleeding in areas where endoscopic operation is difficult, or in the case of diffuse bleeding, tumor bleeding, and recurrent bleeding. Herein, we proposed a novel treatment strategy using biomaterials to protect the wound and isolate the erosion of digestive tract contents to prevent arterial bleeding in advance. By introducing chitosan to construct multihydrogen-bonding and an electrostatic interaction system, we developed polyethyleneimine/polyacrylic acid/chitosan (PEI/PAA/CS) multifunctional hydrogel. The new hydrogel is characterized by ultrafast gelation, strong tissue adhesion, gastric acid resistance, burst resistance, biocompatibility, hemostasis, and tissue repair. The addition of CS significantly improved the tissue adhesion, biocompatibility, hemostasis, and tissue repair ability of the hydrogel. The PEI/PAA/CS hydrogel could adhere to the ulcer surface and form a protective layer on the wound to prevent arterial bleeding. Importantly, the PEI/PAA/CS hydrogel also has the ability to stop bleeding and promote wound repair, which has been demonstrated in a variety of hemorrhage models in rats and rabbits. All of these factors indicate that the PEI/PAA/CS hydrogel is a promising biomaterial for reducing the risk of ulcerative arterial bleeding.

Design and validation of performance-oriented injectable chitosan thermosensitive hydrogels for endoscopic submucosal dissection.

Endoscopic submucosal dissection (ESD) is a challenging procedure. The use of biomaterials to improve the operator's convenience (operating affinity) has received little attention. We prepared two thermosensitive hydrogels, lactobionic acid-modified chitosan/chitosan/ß-glycerophosphate thermosensitive hydrogel (hydrogel 1) and its lyophilized powders (hydrogel 2), characterized their physicochemical properties and evaluated their performance in ESD experiments on large animals, by comparing with the commonly used normal saline (NS) and glycerin fructose (GF). These hydrogels showed good low-temperature fluidity; their viscosities at 4 °C were 92.2 mPa.s and 26.9 mPa.s, respectively. The hydrogels provided significantly better viscoelastic properties than NS and GF. The relaxation moduli of hydrogels were higher than those of NS and GF when the strains were 1 %, 5 %, and 10 %. The hydrogels can be maintained for seven days, even at pH 1, after which they degrade entirely. In pig model experiments, we performed submucosal injection and ESD procedures in the stomach and esophagus. The cushion height produced by the hydrogels was higher than those of NS and GF 30 min after injection. The ESD operation time for hydrogels was significantly shorter. Postoperative wound observation and histological analysis showed that the hydrogels promoted wound healing. The two hydrogels differed in fluidity, viscoelasticity, and other properties, which makes it possible to select the hydrogels according to the size and location of the lesion during ESD operation, and hydrogel 2 may be more suitable for use in lengthier procedures. In general, the hydrogels showed good performance, facilitated the intraoperative operation of ESD, shorten the operation time and promoted wound healing, which is of great significance for reducing the complications and reducing the threshold of ESD operation and further promoting the popularity of ESD.

Nanocomposite hydrogel based on chitosan/laponite for sealing and repairing tracheoesophageal fistula.

Tracheoesophageal fistula (TEF) is an abnormal connection between the trachea and esophagus that severely impairs quality of life. Current treatment options have limitations, including conservative treatment, surgical repair, and esophageal stent implantation. Here, we introduced laponite (LP) nano-clay to improve chitosan-based hydrogels' rheological properties and mechanical properties and developed an endoscopically injectable nanocomposite shear-thinning hydrogel to seal and repair fistulas as an innovative material for the treatment of TEF. Excellent injectability, rheological properties, mechanical strength, self-healing, biodegradability, biocompatibility, and tissue repair characterize the new hydrogel. The introduction of LP nano-clay improves the gel kinetics problem of hydrogels to realize the sol-gel transition immediately after injection, avoiding gel flow to non-target sites. The addition of LA nano-clay can significantly improve the rheological properties and mechanical strength of hydrogels, and hydrogel with LP content of 3 % shows better comprehensive performance. The nanocomposite hydrogel also shows good cytocompatibility and can promote wound repair by promoting the migration of HEEC cells and the secretion of VEGF and FGF. These findings suggest that this nanocomposite hydrogel is a promising biomaterial for TEF treatment.

Hydrogels of collagen/chondroitin sulfate/hyaluronan interpenetrating polymer network for cartilage tissue engineering.

The network structure of a three-dimensional hydrogel scaffold dominates its performance such as mechanical strength, mass transport capacity, degradation rate and subsequent cellular behavior. The hydrogels scaffolds with interpenetrating polymeric network (IPN) structure have an advantage over the individual component gels and could simulate partly the structure of native extracellular matrix of cartilage tissue. In this study, to develop perfect cartilage tissue engineering scaffolds, IPN hydrogels of collagen/chondroitin sulfate/hyaluronan were prepared via two simultaneous processes of collagen self-assembly and cross linking polymerization of chondroitin sulfate-methacrylate (CSMA) and hyaluronic acid-methacrylate. The degradation rate, swelling performance and compressive modulus of IPN hydrogels could be adjusted by varying the degree of methacrylation of CSMA. The results of proliferation and fluorescence staining of rabbit articular chondrocytes in vitro culture demonstrated that the IPN hydrogels possessed good cytocompatibility. Furthermore, the IPN hydrogels could upregulate cartilage-specific gene expression and promote the chondrocytes secreting glycosaminoglycan and collagen II. These results suggested that IPN hydrogels might serve as promising hydrogel scaffolds for cartilage tissue engineering.

[Biocompatibility analysis of hyaluronic acid sodium gels for medical application].

Hyaluronan acid sodium gels are used in ophthalmic surgery, orthopedic treatment and cosmetic surgery. In 2009,there were 12 domestic manufacturers in China producing 33 kinds of products. 23 kinds of imported products were allowed by SFDA to sale in the meantime. Since manufacturers use different production processes, product performances are quite different. According to the GB/T 16886. 1-2001, we designed a pilot program to evaluate the sodium hyaluronate gel products comprehensively in this paper. The results showed that, except chromosome aberration test of gel A and subchronic systemic toxicity of gel C appeared positive, the remaining samples of the test results were negative. This article provides a reference to write standard of cross-linked hyaluronic sodium gel and the revision of standard YY0308-2004.

A simple, safe and easily accessible polyvinyl alcohol hydrogel for wound cleaning.

Acute wounds are often contaminated by some kind of filth, and fluids are usually used to wash away the dirt, but the force of the fluid may cause secondary injury at the wound site or even increase the risk of infection. Hydrogels have several advantages over liquid scouring since they are less intense, more portable, and easier to control. In this study, poly(vinyl alcohol) was used to prepared a series of hydrogels, which were tested in terms of their properties and abilities to clean simulated dirty wounds. Simulated dirt and bacterial (Serratia marcescens) adhesion experiments demonstrated that they could effectively adhere to a certain amount of dirt or bacteria to achieve the purpose of wound cleaning. In addition to the bacterial adhesion, the antibacterial experiments also proved that the hydrogels have a certain inhibitory effect on the proliferation of E.coli and S.aureus. The hydrogels could change shape freely and exhibited excellent biocompatibility, ductility, and self-healing capabilities, which increase their service life and make them more suitable for treating wounds or acting as protection buffers. Based on all these characteristics, the developed hydrogel may be a potentially valuable material for wound cleaning.

Vanillin enhances the antibacterial and antioxidant properties of polyvinyl alcohol-chitosan hydrogel dressings.

Wound management requires the preparation of controllable, safe and effective dressings to isolate the wound from the external environment. Currently, widely used commercial dressings focus on the isolation effect rather than an environment conducive to wound healing. To provide the dressing with beneficial properties such as wetting and antioxidant and antibacterial activity, this study used polyvinyl alcohol (PVA) hydrogel as the base material and introduced chitosan (CS) and vanillin (V) to design a PVA/CS/V three-phase hydrogel dressing. The dressings were prepared using a freeze-thaw cycle to achieve properties. We conducted a comparative analysis of PVA/V and PVA/CS two-phase hydrogels. The PVA/CS/V (PCV) hydrogel dressing maintaining an elastic modulus at >5 kPa at 15-40 °C. An in vitro antibacterial assay showed the potent antibacterial ability of hydrogels against gram-positive and -negative bacteria, and cells in some PCV groups showed higher activity. The antioxidant results showed that PCV hydrogel had a potent scavenging effect on DPPH, ABTS+, and PTIO free radical. The antibacterial and antioxidant properties of three-phase hydrogel showed the best performance in all experimental groups. These results suggest that PCV hydrogel has value in commercial applications due to its simple preparation process and excellent biological properties.

Lactobionic acid-modified chitosan thermosensitive hydrogels that lift lesions and promote repair in endoscopic submucosal dissection.

To develop a biomaterial to lift the lesion and promote wound healing in endoscopic submucosal dissection (ESD), we used lactobionic acid (LA) to improve the water solubility of chitosan (CS) and prepared a new three-phase hydrogel system with lactobionic acid-modified chitosan/chitosan/ß-glycerophosphate (CSLA/CS/GP). The results indicated that the hydrogel retains temperature-sensitive properties, and CSLA obviously improved the low-temperature fluidity of the hydrogel precursor solution, enabling injection of the hydrogel by endoscopic needle. The mechanical strength and bio-adhesion of the hydrogels were also improved by the addition of CSLA and the hydrogels could be maintained in acidic environment for a few days and exhibit greater protection of cells. The CSLA/CS/GP hydrogels show good cytocompatibility. The heights of cushions elevated by CSLA/CS/GP hydrogels remained ∼ 60 % 2 h post-injection in porcine stomach models. Given the unique characteristics of these materials, the CSLA/CS/GP thermo-sensitive hydrogel is a promising intraoperative biomaterial in ESD.

Evaluating platelet activation related to the degradation products of biomaterials using molecular markers.

Increasing numbers of biodegradable medical devices may be used in the circulatory system. The effects of the released degradation products from these medical devices on the blood may be gradual and cumulative. When they reach critical levels, they may cause thrombosis and other complications. For this reason, it is important to evaluate the blood compatibility of degradation products for quality control and development of these devices. In the present study, we evaluated the degradation products of four biodegradable materials (collagen, polylactic acid, calcium phosphate ceramics, and magnesium) using platelet activation molecular markers that are associated with thrombosis. We found that the degradation products activate platelets to a certain extent, and that the degradation products produced during various degradation time periods activate platelets to varying degrees. This platelet activation occurs via several mechanisms, most of which are associated with the physicochemical properties of the degradation products, including ion concentration, pH, molecular microstructure, and molecular weight. Our findings not only provide a clearer understanding of the effects of degradation products from blood-contacting biodegradable devices, but also provide material for screening of degradation behavior so as to improve quality control for these devices.

Feasibility study of use of rabbit blood to evaluate platelet activation by medical devices.

It is important to ascertain platelet responses to blood-contacting medical devices as part of a complete hemocompatibility evaluation. Nevertheless, researchers often face the problem of insufficient quantities of human blood for evaluation of platelet activation by actual medical devices. If animal blood can replace human blood to evaluate platelet activation by medical devices, testing will be smoother and will aid for quality control of related products. Therefore, in this study, we exposed representative biomaterials to human blood, rabbit blood and mouse blood, and evaluated similarities and differences in platelet activation among the three types of blood by measuring various molecular markers. We found that rabbit blood and human blood had considerable similarity in terms of platelet activation, while mouse blood and human blood showed considerable differences. Therefore, rabbit blood may replace human blood for platelet function testing.

[A comparative study of PTT and CT tests for coagulation evaluation of cardiovascular system external communicating devices].

Based on GB/T16886.4-2003 Standard, the coagulation effects of 5 cardiovascular system external communicating devices made consist of metal and polymer were assessed using the partial thromboplastin time (PTT) and Lee-White coagulation time (CT) tests. The results indicate that PTT test is a stable and valuble method for evaluating the coagulating pathway disturbance of the devices. In line with GB/T 16886.4-2003 Standard, PTT test is a recommendable method for evaluating the external communicating devices Based on GB/T14233.2-2005 Standard, CT test is a method more liable to variation, compared with PTT test. This is due to the complex relativity in the test itself, due to all the factors of coagulating cascade, and due to the individual difference of animal. The question of how to select proper negative control for coagulation evaluation of the Cardiovascular System External Communicating Devices in clinical setting for CT test should be addressed and studied.

Evaluating platelet activation related to the degradation of biomaterials using molecular markers.

The effective assessment of platelet activation is an important component of the evaluation of cardiovascular implants. Currently, most evaluation is performed based on the ISO 10993-4 international standard. However, the methods specified in this standard were originally designed for non-degradable materials, and the applicability of these methods to evaluate degradable materials has not been carefully assessed. Here, the platelet activation response was evaluated (using blood from health rabbits) for three typical degradable materials (collagen, polylactic acid, and hydroxyapatite) by measuring the widely used molecular markers CD62 P, CD63, and CD40 L and the three molecular markers PF4, ß-TG, and TXB2 that are referenced in the ISO 10993-4 standard. The variations of these six markers were compared in the simulated degradation of the three test materials. The results showed differences in platelet activation with degradation that were strongly related to the surface physicochemical properties. Changes in the surface roughness and contact angle of the materials correlated with changes in the degree of platelet activation. The six tested platelet activation molecular markers show promise for assessment of platelet function in degradable medical devices, providing guidance for quality control strategies and the design and improvement of safe medical devices.

Hydroxypropylcellulose enhanced high viscosity endoscopic mucosal dissection intraoperative chitosan thermosensitive hydrogel.

In order to prepare a submucosal injection agent that can gel during the endoscopic mucosal dissection (ESD) and still adhere to the wound surface after the surgery to reduce intraoperative and postoperative complications, a thermosensitive hydrogel based on chitosan/ß-glycerophosphate (CS-GP) was constructed. Hydroxypropylcellulose (HPC) and collagen Ⅰ were added into the hydrogel to improve the adhesion ability, viscosity and biocompatibility. The effect of HPC on the viscosity, strength and bio-adhesion and collagen Ⅰ on the biocompatibility and promoting on the wound restoration were intensively studied. The results indicated that HPC and collagen Ⅰ will not obviously change pH of the system and the system was still physically cross-linked. HPC increased the initial gelation time but the time of the entire gelation process is shortened, especially the high-concentration HPC, and improved the mechanical strength obviously. The viscosity and the bio-adhesion of the CS-GP hydrogels were improved by the addition of HPC. Also, the mixed collagen Ⅰ offered good cytocompatibility. According to the results of cultured GES-1 cell, it can be seen that the addition of HPC and collagen Ⅰ does not inhibit cell proliferation, and shows more obvious protection on the cells under acidic conditions. All of these suggested that the application of this hydrogel were more promising in ESD surgery than which without HPC and collagen Ⅰ.

Fabrication and characterization of collagen-based injectable and self-crosslinkable hydrogels for cell encapsulation.

Injectable and self-crosslinkable hydrogels have drawn much attention for their potential application as cell delivery carriers to deliver cells to the injury site of arbitrary shape. In this study, injectable and self-crosslinkable hydrogels were designed and fabricated based on collagen type I (Col I) and activated chondroitin sulfate (CS-sNHS) by physical and chemical crosslinking without the addition of any catalysts. The physical properties of hydrogels, including mechanical properties, swelling and degradation properties, were investigated. The results demonstrated that the physical properties of hydrogels, especially the stiffness of hydrogels, were readily tuned by varying the degree of substitution (DS) of CS-sNHS without changing the concentration of collagen-based precursor. Chondrocytes were encapsulated into hydrogels to investigate the effects of hydrogels on the survival, proliferation and extracellular matrix (ECM) secretion of cells by FDA/PI staining, CCK-8 test and histological staining. The results suggested that all of these hydrogels supported the survival and ECM secretion of chondrocytes, while there was more ECM secretion around chondrocytes encapsulated in hydrogel Col I/CS-sNHS56% in which the DS of CS-sNHS was 56%. When the neutral precursor solution for hydrogel of Col I or Col I/CS-sNHS56% was subcutaneously injected into SD rats, hydrogels both displayed acceptable biocompatibility in vivo. These results imply that these injectable and self-crosslinkable hydrogels are suitable candidates for applications in the fields of cell delivery and tissue engineering.

A novel biomimetic composite substitute of PLLA/gelatin nanofiber membrane for dura repairing.

OBJECTIVE: Biomimetic design will significantly improve growth and regeneration of dural cells and tissue for better repairing effects and fewer complications in repairing the native dura. This study designed a novel composite, biomimetic substitute based on the characteristics of native dura extracellular matrix. METHODS AND RESULTS: This substitute is expected to rapidly induce cell adhesion, migration, and fast regeneration of neotissue. The material characteristics (contact angle, surface charge, and zeta potential were evaluated), in vitro biological characteristics (cell stretch, connections between cells, cell proliferation) and in vivo tissue regeneration capability of this substitute were evaluated, compared to those of collagen dura substitute, the mostly used dura substitute. The results showed that the surface properties of this composite substitute were more biomimetic to native extracellular matrix than collagen substitute did, together with better cytocompatibility, tissue ingrowth, and neoangiogenesis. This composite substitute further demonstrated in clinical case study its ideal repair effect with no CSF leakage or other adverse reactions. CONCLUSION: In conclusion, the new biomimetic composite substitute provides alternative substitute for dura repairing.

Effect of phase composition on protein adsorption and osteoinduction of porous calcium phosphate ceramics in mice.

The purpose of this study was to investigate the effect of phase compositions of porous calcium phosphate (CaP) ceramics on their protein adsorption behaviors in vitro and osteoinductive potentials in vivo in mice. Under competitive conditions, a high adsorption of bone morphogenetic protein 2 (BMP-2) was observed at a high initial concentration of BMP-2 in the multi-protein solution on all the four types of ceramics, indicating their strong affinity for BMP-2. No significant difference in BMP-2 adsorption between the ceramics was noted, indicating that phase composition could have little influence on BMP-2 adsorption. After implantation into the thigh muscles of mice for 45 and 90 days, the histological and histomorphometric analyses showed that porous biphasic calcium phosphate (BCP) ceramic consisting of 30% hydroxyapatite HA and 70% tricalcium phosphate (ß-TCP), i.e. BCP-2 had stronger osteoinductive ability than the other three groups of ceramics. The immunohistochemical staining showed the highest expression of BMP-2 and osteocalcin (OCN) in BCP-2 group. Osteoinduction of porous CaP ceramics might be influenced by the amount of BMP-2 present in the local microenvironment in the implant, which was regulated by the phase composition of the ceramics. BCP-2 promoted the highest expression of BMP-2 and then showed the strongest osteoinduction in mice.