In vivo evaluation of a novel alginate dressing.

Alginate dressings are currently used in the management of epidermal and dermal wounds, and provide a moist environment that leads to rapid granulation and reepithelialization. However, a cytotoxic effect on proliferation of fibroblasts and residual material with inflammation in healing wounds have been reported recently. We have developed a new alginate dressing (AGA-100), which does not have an inhibitory effect on proliferation of fibroblasts. The purpose of this study was to evaluate the new alginate dressing with respect to wound healing in full- and partial-thickness pig wounds and with respect to biodegradation following implantation into rabbit muscle. Kaltostat and Sorbsan, both well-established commercial dressings, were used as control. The closure rate of full-thickness wounds treated with AGA-100 was significantly higher on day 15 compared with that with Kaltostat and Sorbsan. Reepithelialization rate of partial-thickness wounds treated with Sorbsan was statistically significantly lower on day 3 than those with the other two dressings. As to dressing debris remained in the healing wound, a large amount of foreign debris was noted in all the full-thickness wounds treated with Kaltostat or Sorbsan, while only about one-third of wounds treated with AGA-100 showed a little dressing debris. AGA-100 implanted into the muscle of rabbits was bioresorbed completely within 3 months. Therefore, dressing residue in AGA-100-treated full-thickness wounds might be fully absorbed in a few months. In conclusion, it is shown that our newly developed AGA-100 possesses superior properties compared with typical alginate dressings.     

In vitro and in vivo evaluation of the chitosan/Tur composite film for wound healing applications

We have developed tourmaline/chitosan (Tur/CS) composite films for wound healing applications. The characteristics of composite films were studied by optical microscope, infrared spectra and X-ray diffraction. Tur particles were uniformly distributed in the CS film and the crystal structure of CS was not remarkably changed except the decrease of crystallinity. The influence of Tur on wound healing applications was characterized by modulating Tur concentrations in the Tur/CS composite film prepared by loading Tur powder into CS matrix with different proportion (0, 1/40 and 1/10). Then L929 cells were co-cultured on the composite films to access the cytotoxicity in vitro. Tur concentrations strongly influenced cell process extension. Tur/CS composite film with 1/40 mass ratio could promote the cell adhesion and proliferation. Fewer and shorter processes were observed at high Tur density. When the composite films were transplanted on porcine full-thickness burn wounds, histological results demonstrated that the Tur/CS group with 1/40 mass ratio had a significantly higher number of newly-formed and mature blood vessels, and fastest regeneration of dermis. Based on the observed facts these films can be tailored for their potential utilization in wound healing and skin tissue engineering applications.     

Chitosan-alginate PEC membrane as a wound dressing: Assessment of incisional wound healing

Flexible, thin, transparent, novel chitosan-alginate polyelectrolyte complex (PEC) membranes, cast from aqueous suspensions of chitosan-alginate coacervates with CaCl(2), were evaluated as potential wound-dressing materials. MTT and NR assays suggested that the chitosan-alginate PEC membranes and their aqueous extracts were nontoxic towards mouse and human fibroblast cells. Cell growth was also not hindered by co-incubation with the membranes. Compared to conventional gauze dressing, the PEC membranes caused an accelerated healing of incision wounds in a rat model. Wounds closed at 14 days postoperatively, and histological observations showed mature epidermal architecture with keratinized surface of normal thickness and a subsided inflammation in the dermis. This was followed by an excellent remodeling phase with organized thicker collagen bundles and mature fibroblasts at 21 days postoperative. Control wounds continued to show signs of an active inflammatory phase under scab on Day 21. Closure rate and appearance of PEC membrane-treated wounds were comparable with Opsite(R)-treated wounds. On the basis of its biocompatibility and wound-healing efficacy, the chitosan-alginate PEC membrane can be considered for wound-dressing applications.     

Electrospun nanofibrous polyurethane membrane as wound dressing

Produced via electrospinning, polyurethane membrane, which has a unique property, has been of interest in medical fields. Electrospinning is a process by which nanofibers can be produced by an electrostatically driven jet of polymer solution. Electrospun fibers are collected in the form of membranes. The porous structured electrospun membrane is particularly important for its favorable properties: it exudates fluid from the wound, does not build up under the covering, and does not cause wound desiccation. The electrospun nanofibrous membrane shows controlled evaporative water loss, excellent oxygen permeability, and promoted fluid drainage ability, but still it can inhibit exogenous microorganism invasion because its pores are ultra-fine. Histological examination indicates that the rate of epithelialization is increased and the dermis becomes well organized if wounds are covered with electrospun nanofibrous membrane. This electrospun membrane has potential applications for wound dressing based upon its unique properties.     

Norfloxacin-loaded chitosan sponges as wound dressing material

The aim of this study was the preparation and characterization of chitosan sponges including a model antibiotic (i.e., norfloxacin). The chitosan sponges were prepared by a solvent evaporation method. The matrix was also cross-linked during the preparation. The results indicated that the chitosan sponges were in the fibrillar structure. The swelling behavior, norfloxacin loading, in vitro release characteristics, and antibacterial activity were determined. The effects of cross-linker concentration, norfloxacin/chitosan ratio, chitosan molecular weight, and base concentration were investigated. The most effective parameter was found to be the degree of neutralization. It was also observed that the equilibrium swelling ratio decreased with increasing cross-linking density. The norfloxacin release was found to be swelling controlled initially and diffusion controlled at the extended release periods. It was also found that the antibacterial activity was directly proportional to the release rate.     

Systematic review of the use of honey as a wound dressing

Objective  

To investigate topical honey in superficial burns and wounds though a systematic review of randomised controlled trials.     

Preparation and characterization of chitin beads as a wound dressing precursor

Chitin was dissolved in N, N-dimethylacetamide/5% lithium chloride (DMAc/5%LiCl) to form a 0.5% chitin solution. Chitin beads were formed by dropping the 0.5% chitin solution into a nonsolvent coagulant, ethanol. The beads were left in ethanol for 24 h to permit hardening, consolidation, and removal of residual DMAc/5%LiCl solvent in order to give spherical chitin beads uniform size distribution. The ethanol-gelled chitin beads had an average diameter of 535 microm. The chitin beads were subsequently activated in 50% (w/v) NaOH solution and reacted with 1.9 M monochloroacetic acid/2-propanol solution to introduce a carboxymethylated surface layer to the chitin beads. The bilayer character of the surface-carboxymethylated chitin (SCM-chitin) beads was verified by Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), and confocal microscopy. The bilayered SCM-chitin beads were found to absorb up to 95 times their dry weight of water. These SCM-chitin beads have potential as a component of wound dressings.     

Fabrication and characterization of a sponge-like asymmetric chitosan membrane as a wound dressing

A novel asymmetric chitosan membrane has been prepared by immersion-precipitation phase-inversion method and evaluated as wound covering. This new type of chitosan wound dressing which consists of skin surface on top-layer supported by a macroporous sponge-like sublayer was designed. The thickness of the dense skin surface and porosity of sponge-like sublayer could be controlled by the modification of phase-separation process using per-evaporation method. The asymmetric chitosan membrane showed controlled evaporative water loss, excellent oxygen permeability and promoted fluid drainage ability but could inhibit exogenous microorganisms invasion due to the dense skin layer and inherent antimicrobial property of chitosan. Wound covered with the asymmetric chitosan membrane was hemostatic and healed quickly. Histological examination confirmed that epithelialization rate was increased and the deposition of collagen in the dermis was well organized by covering the wound with this asymmetric chitosan membrane. The results in this study indicate that the asymmetric chitosan membrane thus prepared could be adequately employed in the future as a wound dressing.     

Development of an infection-resistant, bioactive wound dressing surface

Trauma, whether caused by an accident or in an intentional manner, results in significant morbidity and mortality. The goal of this study was to develop a novel biomaterial surface in vitro and ex vivo that provides both localized infection resistance nd hemostatic properties. Our hypothesis is that a combination of specific surface characteristics can be successfully incorporated into a single biomaterial. Functional groups were created with woven Dacron (Cntrl) material via exposure to ethylenediamine (C-EDA). The antibiotic ciprofloxacin (Cipro) was then applied to the C-EDA material using pad/autoclave technique (C-EDA-AB) followed by surface immobilization of the coagulation cascade enzyme thrombin (C-EDA-AB-Thrombin). Antimicrobial activity by the C-EDA-AB surface persisted for 5 days compared with Cntrl and dipped controls, which lasted <1 h. C-EDA-AB-Thrombin surfaces had 2.6- and 105-fold greater surface thrombin activity compared with nonspecifically bound thrombin and Cipro-dyed surfaces, respectively. Surface thrombus formation ex vivo was evident after 1 min of exposure, with thrombus organization evident by 2.5 min. In contrast, C-EDA-AB and Cntrl segments showed only blood protein adsorption on the fibers. Thus, this study demonstrated that Cipro and thrombin can be simultaneously incorporated onto a biomaterial surface while maintaining their respective biological activities.     

Ethyl acetate Salix alba leaves extract-loaded chitosan-based hydrogel film for wound dressing applications

High toxicity and multidrug resistance associated with various standard antimicrobial drugs have necessitated search for safer alternatives in plant-derived materials. In this study, we performed biological examination of chitosan-based hydrogel film loaded with ethyl acetate Salix alba leaves extract against 11 standard laboratory strains. FTIR showed regeneration of saccharide peak in CP1A at 1047 cm^?1 and increased in height of other peaks. DSC exothermic decomposition peaks at 112 °C, 175 °C and 251 °C reveal the effect of extract on hydrogel film. From FESEM images, three-dimensional cross-linking and extract easily seen in the globular form from the surface. MTT assay on HEK 293 cells showed that CP1A was non-toxic. Minimum inhibitory concentration ranges from 4000 μg/ml to 125 μg/ml. Enterococcus faecium, Candida glabrata and Candida tropicalis were the most resistant, while Salmonella typhi and Candida guilliermondii were the most susceptible micro-organisms.     

Biofunctionalized electrospun silk mats as a topical bioactive dressing for accelerated wound healing

Materials able to deliver topically bioactive molecules represent a new generation of biomaterials. In this article, we describe the use of silk mats, made of electrospun nanoscale silk fibers containing epidermal growth factor (EGF), for the promotion of wound healing processes. In our experiments, we demonstrated that EGF is incorporated into the silk mats and slowly released in a time-dependent manner (25% EGF release in 170 h). We tested these materials using a new model of wounded human skin-equivalents displaying the same structure as human skin and able to heal using the same molecular and cellular mechanisms found in vivo. This human three-dimensional model allows us to demonstrate that the biofunctionalized silk mats, when placed on the wounds as a dressing, aid the healing by increasing the time of wound closure by the epidermal tongue by 90%. The preservation of the structure of the mats during the healing period as demonstrated by electronic microscopy, the biological action of the dressing, as well as the biocompatibility of the silk demonstrate that this biomaterial is a new and very promising material for medical applications, especially for patients suffering from chronic wounds.     

Spandra: a sustained release battlefield wound dressing

In 1981, our laboratories developed a family of elastomers which could be cured by ultraviolet (UV) radiation. Curing by UV radiation was a significant advance in chemistry, since it allowed ultra-fast curing of elastomers in a matter of seconds, as compared to several hours at 110 degrees C for conventional heat curing. We applied for a patent based on this technology, and the patent was allowed in mid-1984 [29]. Based on this technology, Thermedics submitted a proposal to the US Army for the development of a sustained-release battlefield wound dressing containing antibiotics and coagulants. The drugs were evenly distributed in the oligomer matrix, and subsequently cured in seconds under UV illumination, without the use of heat, organic solvents or water. Because delicate drugs are not subjected to heat, organic solvents or water, the pharmacological activity of the drugs is insured. Therefore, theoretically any drug may be incorporated into our dressing. Sustained release dressings were first developed at Thermedics in 1983, spurred by a contract from the US Army Medical Research and Development Command. Under this contract, the Company developed a new type of wound dressing capable of accelerating the healing process, retarding infection, and minimizing pain. Based on our TECOFLEX materials technology, the dressing performs like temporary artificial skin. Its transmission properties for oxygen, carbon dioxide, and water vapor are similar to those of intact skin. Thus, while excluding bacteria from the wound site, the dressing maintains an optimal moist environment for the promotion of rapid healing. The new drawing shown in Figure 10 minimizes pain during healing by preventing dehydration and shrinkage in the wound. Patient comfort is also enhanced by the incorporation of a special fabric which imparts flex properties to the bandage that are almost identical to those of human skin, with greater stretch in one direction than in another. This also facilitates application to complex body contours by only one attendant, an important feature in both hospital and emergency situations. Materials currently in use in hospitals are difficult to handle, requiring two or three nurses to apply large dressings. Thermedics' military wound dressing not only has the significant advantage of ease of application, but this dressing can also be used for delivery of drugs to a specific site.(ABSTRACT TRUNCATED AT 400 WORDS)     

In vitro and in vivo evaluation of ultrananocrystalline diamond as an encapsulation layer for implantable microchips

Thin ultrananocrystalline diamond (UNCD) films were evaluated for use as hermetic and bioinert encapsulating coatings for implantable microchips, where the reaction to UNCD in vitro and in vivo tissue was investigated. Leakage current tests showed that depositing UNCD coatings, which were conformally grown in (1% H₂) Ar/CH₄ plasma, on microchips rendered the surface electrochemically inactive, i.e. with a very low leakage current density (2.8 × 10⁻⁵ A cm⁻² at −1 V and 1.9 × 10⁻³ A cm⁻² at ± 5 V) ex vivo. The impact of UNCD with different surface modifications on the growth and activation of macrophages was compared to that of standard-grade polystyrene. Macrophages attached to oxygen-terminated UNCD films down-regulated their production of cytokines and chemokines. Moreover, with UNCD-coated microchips, which were implanted subcutaneously into BALB/c mice for up to 3 months, the tissue reaction and capsule formation was significantly decreased compared to the medical-grade titanium alloy Ti–6Al–4V and bare silicon. Additionally, the leakage current density, elicited by electrochemical activity, on silicon chips encapsulated in oxygen-terminated UNCD coatings remained at the low level of 2.5 × 10⁻³ A cm⁻² at 5 V for up to 3 months in vivo, which is half the level of those encapsulated in hydrogen-terminated UNCD coatings. Thus, controlling the surface properties of UNCDs makes it possible to manipulate the in vivo functionality and stability of implantable devices so as to reduce the host inflammatory response following implantation. These observations suggest that oxygen-terminated UNCDs are promising candidates for use as encapsulating coatings for implantable microelectronic devices.     

聚己内酯/丝胶蛋白静电混纺膜炎症响应的体内评价

背景：研究表明,丝胶蛋白可以增加哺乳动物细胞的增殖,促进伤口愈合,具有良好的抗菌、抗氧化、抗癌的活性,同时具有良好的生物相容性和可降解性,是一种理想的组织工程材料。然而目前对丝胶蛋白能否引起炎症响应仍存在争论。 目的：评价聚己内酯/丝胶蛋白静电混纺膜的炎症响应。 方法：将不同混合比例的聚己内酯/丝胶蛋白纳米纤维薄膜体内埋植入大鼠背部肌肉,4周后取出进行组织学观察,通过苏木精-伊红染色和巨噬细胞抗体CD68免疫组织化学染色评价其炎症响应。 结果与结论：聚己内酯/丝胶蛋白7∶3组混纺膜炎症反应与丝素蛋白膜类似,小于聚己内酯/丝胶蛋白6∶4组、聚己内酯/丝胶蛋白5∶5组和聚己内酯组;聚己内酯/丝胶蛋白6∶4组与聚己内酯/丝胶蛋白5∶5组与聚己内酯组类似。提示聚己内酯/丝胶蛋白静电混纺膜不会引起显著的炎症响应,少量添加丝胶蛋白可以降低聚己内酯的炎症反应,随着丝胶蛋白含量的增加,炎症反应呈上升趋势。     

Chemical and physical properties of a hydrogel wound dressing

Geliperm hydrogel provides optimal physiological conditions for wound healing. The material is composed of two interlaced networks, one of polyacrylamide and one of agar, and contains about 96% firmly bound water. It is supplied in smooth, elastic, transparent sheets which are impermeable to bacteria but permeable to gases, salts, metabolites and proteins. Geliperm is nontoxic and has no irritative properties. Mechanical properties, water retention and diffusion of dyes and proteins are reported. Bacterial size should preclude penetration of the gel. The hydrogel in granular form represents a coherent material which could be used in deep fissured wounds and for the treatment of injuries with a large amount of exudation and contamination.     

Sequential antibiotic and growth factor releasing chitosan-PAAm semi-IPN hydrogel as a novel wound dressing

The aim of this study is to prepare a novel wound dressing material which provides burst release of an antibiotic in combination with sustained release of growth factor delivery. This might be beneficial for the prevention of infections and to stimulate wound healing. As a wound dressing material, the semi-interpenetrating network (semi-IPN) hydrogel based on polyacrylamide (PAAm) and chitosan (CS) was synthesized via free radical polymerization. Ethylene glycol dimethacrylate was used for cross-linking of PAAm to form semi-IPN hydrogel. The hydrogel shows high water content (～1800%, in dry basis) and stable swelling characteristics in the pH range of the wound media (～4.0–7.4). The antibiotic, piperacillin–tazobactam, which belongs to the penicillin group was loaded into the hydrogel. The therapeutic serum dose of piperacillin–tazobactam for topic introduction was reached at 1st hour of the release. Additionally, in order to increase the mitogenic activity of hydrogel, epidermal growth factor (EGF) was embedded into the CS–PAAm in different amounts. Cell culture studies were performed with L929 mouse fibroblasts and the simulated cell growth was investigated by 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide assay. The successful sustained release behavior of CS–PAAm hydrogel for EGF maintained the presence of EGF in the culture up to 5?days and the highest mitochondrial activities were recorded for the 0.4?μg EGF-loaded/mg of hydrogel group. In conclusion, CS–PAAm semi-IPN hydrogel loaded with piperacillin–tazobactam and EGF could be proposed for an effective system in wound-healing management.     

In vivo biocompatibility and stability of a substrate-supported polymerizable membrane-mimetic film

The cell membrane establishes an important paradigm for the molecular engineering of coatings for implantable devices because of its intrinsic biocompatibility and ability to act as a template for the assembly of diverse membrane-associated macromolecules. A stabilized membrane-mimetic film was assembled on alginate/Ca(2+) hydrogel microcapsules by in situ polymerization of an acrylate functionalized phospholipid. The phospholipid monomer was prepared as unilamellar vesicles and fused onto octadecyl chains that were components of an amphiphilic terpolymer anchored onto a polyelectrolyte multilayer (PEM) by electrostatic interactions. Microcapsules coated with a membrane-mimetic film were implanted into the peritoneal cavity of C57BL/6 mice, and the short-term biostability and biocompatibility of membrane-mimetic films assembled on two different alginate/poly(l-lysine) PEM cushions were compared. The nature of the underlying PEM support had a profound impact on the biocompatibility of the membrane-mimetic film, as the percentage of retrieved microcapsules completely overgrown with host cells shifted from 66+/-5.9% to less than 1% when modifications to the PEM were made. When assembled on the appropriate PEM support, biocompatibility of membrane-mimetic-coated microspheres was high wherein 87.5+/-5.7% of the implanted microspheres were retrieved 4 weeks after implantation and 92.6+/-6.4% of the retrieved capsules were free of cell adhesion or fibrotic overgrowth. Finally, 4 weeks after implantation, microspheres coated with a Texas red-labeled membrane-mimetic film were imaged with confocal microscopy and exhibited a uniform film around the periphery of the implant, indicating a high degree of film biostability. Hence, membrane-mimetic films provide a new route for generating robust, biocompatible, and biochemically heterogeneous coatings for implantable devices through molecular self-assembly.