Human neutrophil elastase and collagenase sequestration with phosphorylated cotton wound dressings

The design and preparation of wound dressings that redress the protease imbalance in chronic wounds is an important goal of wound healing and medical materials science. Chronic wounds contain high levels of tissue and cytokine-destroying proteases including matrix metalloprotease and neutrophil elastase. Thus, the lowering of excessive protease levels in the wound environment by wound dressing sequestration prevents the breakdown of extracellular matrix proteins and growth factors necessary for wound healing. Phosphorylated cotton wound dressings were prepared to target sequestration of proteases from chronic wound exudate through a cationic uptake binding mechanism involving salt bridge formation of the positively charged amino acid side chains of proteases with the phosphate counterions of the wound dressing fiber. Dressings were prepared by applying sodium hexametaphosphate and diammonium phosphate in separate formulations to cotton gauze by pad/dry/cure methods. Phosphorylated cotton dressings were assessed for their ability to lower elastase and collagenase activity. The phosphorylated cotton dressings lowered elastase and collagenase activity 40-80% more effectively than the untreated cotton wound dressings under conditions that mimic chronic wound exudate. Efficacy of the phosphorylated cotton was found to be related to the level of phosphorylation and a lower pH due to protonated phosphate at the surface of the dressing. The capacity of the modified gauze to sequester continued elastase secretions similar to that found in a chronic wound over a 24-h period was retained within a 80% retention of elastase sequestration and was dose-dependent.     

In vitro and in vivo assays on egg white/polyvinyl alcohol/clay nanocomposite hydrogel wound dressings

Novel nanocomposite hydrogel wound dressings on the basis of egg white and polyvinyl alcohol, as matrix, and natural Na-montmorillonite clay, as reinforcing agent, were prepared and their performances on wound healing investigated in vitro and in vivo. In vitro cytotoxicity assay revealed non-cytotoxic activity and excellent biocompatibility level of prepared nanocomposite hydrogel wound dressings. The bacterial penetration assay showed the prepared nanocomposite hydrogel wound dressings are excellent barriers against microorganisms and could protect the wound from infection during the wound healing. In vivo animal study showed that the wound healing process was considerably faster in wounds covered with nanocomposite hydrogel wound dressings compared to the conventional wound dressing, i.e. sterile gauze, due to creation of a moist environment on the wound surface and faster migration rate of the epidermal cells. The mechanical properties of healed wounds with nanocomposite hydrogel wound dressings were better than those control wounds covered with sterile gauze due to their better collagen formation ability as a result of created moist healing condition as well as the presence of egg white, as a source of proteins, in their structures.     

Androgens influence expression of matrix proteins and proteolytic factors during cutaneous wound healing

Excessive proteolytic activity is a feature of chronic wounds such as venous ulcers, in which resolution of the inflammatory response fails and restorative matrix accumulation is delayed as a consequence. The inflammatory actions of native androgens during the healing of acute skin wounds have lately been characterized. We have now investigated the hypothesis that such activities may impact upon the balance between anabolic and catabolic processes during wound healing. We report that wound deposition of both type I collagen and fibronectin is increased in castrated rats compared with control animals. This response is accompanied by early increases and later decreases in overall wound levels of the key collagenolytic enzymes, matrix metalloproteinase (MMP)-1 and MMP-13. Moreover, the activities of MMP-2 and MMP-9, two further enzymes that contribute to collagen digestion during venous ulceration, were significantly decreased in the wounds of castrated rats. Additional analyses provide evidence that androgens directly stimulate dermal fibroblast collagen production, which supports the suggestion that increased wound collagen deposition in androgen-deprived rats results from reduced matrix degradation (as opposed to enhanced matrix protein biosynthesis). Androgen-mediated dysregulation of the parallel processes of collagen deposition and turnover may underscore the delayed healing of cutaneous wounds in elderly male patients and further contribute to the increased incidence of non-healing wounds in this population.     

The inhibition of matrix metalloproteinase activity in chronic wounds by a polyacrylate superabsorber

Excessive matrix metalloproteinase (MMP) levels have been observed in wound fluid of impaired healing wounds. This is thought to interfere with granulation tissue formation as newly formed extracellular matrix and cytokines are degraded and the wound becomes deadlocked, unable to progress to the next healing stages. In the cleansing phase, associated with high MMP activity levels, hydroactive wound dressings containing polyacrylate superabsorber particles are particularly effective. We tested whether these particles can block MMP activity in wound fluid obtained from chronic venous leg ulcers. Polyacrylate superabsorber particles inhibited MMP activity by more than 87% in a fluorogenic peptide substrate assay. Further analysis revealed two underlying molecular mechanisms. First, experiments showed direct binding of MMPs to the particles. Secondly, polyacrylate superabsorber particles can bind Ca2+ and Zn2+ ions competing with MMPs for divalent ions required for enzymatic activity. Furthermore, we provide the first evidence in vivo that MMPs bind effectively to polyacrylate superabsorber particles within the hostile environment of chronic wounds. We conclude that polyacrylate superabsorber particles can rescue the highly proteolytic microenvironment of non-healing wounds from MMP activity so that more conductive conditions allow healing to proceed.     

Accelerating effects of chitosan for healing at early phase of experimental open wound in dogs.

Chitosan is a polymeric beta(1 --> 4) glucosamine (2-amino-2-deoxy-D-glucose) and N-acetyl-D-glucosamine (2-acetamido-2-deoxy-D-glucose) which has been reported as a wound healing accelerator. In order to evaluate the efficacy of chitosan as an accelerator of wound healing, experimental open skin wounds were made on the dorsal side in three normal beagles. Cottonfiber-type chitosan (degree of acetylation = 18%) was applied for 15 days, and the process of wound healing was evaluated histologically and immunohistochemically. On day 3 postwounding, the chitosan-treated wounds showed histologically severe infiltration of polymorphonuclear (PMN) cells and an increase in effusion compared with that in the control. Granulation was more pronounced by the chitosan treatment on day 9 and 15 postwounding. Immunohistochemical typing of collagen I, III and IV showed increase of the production of type III collagen in the chitosan group. The appearance of mitotic cells occurred numerously in the control on postwounding day 3, and in the chitosan group on postwounding day 6. These results suggest chitosan to be having a function in the acceleration of infiltration of PMN cells at the early stage of wound healing, followed by the production of collagen by fibroblasts.     

Alginates from wound dressings activate human macrophages to secrete tumour necrosis factor-alpha

Alginates are used to manufacture a number of wound dressings. Clinical observations indicate that they may initiate or accelerate healing of chronic wounds after treatment of underlying pathology. Wound granulation tissue contains large numbers of macrophages and they are thought to regulate the healing process. As purified alginates have been demonstrated to activate macrophages this study was initiated to determine whether alginates present within wound dressings may interact with wound macrophages. Alginate fibres taken from four commercially available dressings were co-cultured with the human histiocytic lymphoma cell line U937 following its differentiation with PMA. Activation was assessed by measurement of TNFalpha production. Two of the dressings, Seasorb and Tegagen, had a minimal effect whilst Sorbsan at 1 mg/ml induced 302 + 19 pg/ml TNFalpha. This effect was inhibited by polymyxin B indicating that activation was due to endotoxin contamination. Kaltostat induced production of 839 + 36 pg/ml TNFalpha. This effect was induced both by polymyxin inhibitable endotoxin and a direct interaction with the alginate fibres. These data indicate that some alginate containing dressings have the potential to activate macrophages within the chronic wound bed and generate a pro-inflammatory signal which may initiate a resolving inflammation characteristic of healing wounds.     

Biotinylated GHK peptide incorporated collagenous matrix: A novel biomaterial for dermal wound healing in rats

Matrikines are small peptide fragments of extracellular matrix proteins that display potent tissue repair activities. Difficulties in achieving sustained delivery of bioactive concentration of matrikines in the affected area limits their therapeutic use. The present study evaluates the effects biotinylated matrikine peptide (bio-glycyl-histidyl-lysine) incorporated collagen membrane for dermal wound healing processes in rats. Biotinylated peptide incorporated collagen matrix (PIC) showed better healing when compared to wounds treated with collagen matrix [CF (collagen film)] and without collagen [CR (control)]. Binding studies indicate that biotinylated GHK (Bio-GHK) binds effectively to the collagen matrix and red blood cell (RBC) membrane when compared with t-butyloxycarbonyl substituted GHK (Boc-GHK). Wound contraction, increased cell proliferation, and high expression of antioxidant enzymes in PIC treated group indicate enhanced wound healing activity when compared to CF and CR groups. Interestingly Bio-GHK incorporated collagen increases the copper concentration by ninefold at the wound site indicating the wound healing property of Bio-GHK can also be linked with both copper localization and matrikine activities. These results demonstrate the possibility of using Bio-GHK incorporated collagen film as a therapeutic agent in the wound healing process.     

Chromatographic and traditional albumin isotherms on cellulose: a model for wound protein adsorption on modified cotton

Albumin is the most abundant protein found in healing wounds. Traditional and chromatographic protein isotherms of albumin binding on modified cotton fibers are useful in understanding albumin binding to cellulose wound dressings. An important consideration in the design of cellulosic wound dressings is adsorption and accumulation of proteins like albumin at the solid-liquid interface of the biological fluid and wound dressing fiber. To better understand the effect of fiber charge and molecular modifications in cellulose-containing fibers on the binding of serum albumin as observed in protease sequestrant dressings, albumin binding to modified cotton fibers was compared with traditional and chromatographic isotherms. Modified cotton including carboxymethylated, citrate-crosslinked, dialdehyde and phosphorylated cotton, which sequester elastase and collagenase, were compared for their albumin binding isotherms. Albumin isotherms on citrate-cellulose, cross-linked cotton demonstrated a two-fold increased binding affinity over untreated cotton. A comparison of albumin binding between traditional, solution isotherms and chromatographic isotherms on modified cellulose yielded similar equilibrium constants. Application of the binding affinity of albumin obtained in the in vitro protein isotherm to the in vivo wound dressing uptake of the protein is discussed. The chromatographic approach to assessment of albumin isotherms on modified cellulose offers a more rapid approach to evaluating protein binding on modified cellulose over traditional solution approaches.     

Testing intelligent wound dressings.

While occlusive wound dressings help provide patients with moist wound healing to reduce pain and increase reepithelialization rate, the moisture vapor transmission rate (MVTR) of these dressings remains constant even though wound exudate levels may vary with time and from wound to wound. The clinician is therefore faced with exudate buildup in heavily exuding wounds and desiccation in lightly exuding wounds-a situation requiring frequent patient monitoring and dressing changes. Am "intelligent" wound dressing would have the ability to automatically respond to a wound's exudate level by self-adjusting its MVTR to maintain a constant moist wound environment. Such a dressing could help ensure that exudate buildup or wound desiccation is reduced or avoided. Three commercial wound dressings (hydrocolloid, thin film, and membrane laminate) were studied for their ability to alter their MVTR in response to varying moisture level. An efficient test methodology and experimental design was developed, which involved direct and indirect fluid contact with the dressings using two temperatures and two test methods. One dressing, a membrane laminate, was found to exhibit intelligent MVTR behavior. Data is presented which shows this dressing's ability to adjust its MVTR nearly eight-fold as a function of hydration level. Information regarding the mechanism of action of this intelligent dressing is also presented.     

Chitosan-based dressings loaded with neurotensin—an efficient strategy to improve early diabetic wound healing

One important complication of diabetes mellitus is chronic, non-healing diabetic foot ulcers (DFUs). This study aims to develop and use dressings based on chitosan derivatives for the sustained delivery of neurotensin (NT), a neuropeptide that acts as an inflammatory modulator in wound healing. Three different derivatives, namely N-carboxymethyl chitosan, 5-methyl pyrrolidinone chitosan (MPC) and N-succinyl chitosan, are presented as potential biomaterials for wound healing applications. Our results show that MPC has the best fluid handling capacity and delivery profile, also being non-toxic to Raw 264.7 and HaCaT cells. NT-loaded and non-loaded MPC dressings were applied to control/diabetic wounds to evaluate their in vitro/in vivo performance. The results show that the former induced more rapid healing (50% wound area reduction) in the early phases of wound healing in diabetic mice. A NT-loaded MPC foam also reduced expression of the inflammatory cytokine TNF-α (P < 0.001) and decreased the amount of inflammatory infiltrate on day 3. On day 10 MMP-9 was reduced in diabetic skin (P < 0.001), significantly increasing fibroblast migration and collagen (COL1A1, COL1A2 and COL3A1) expression and deposition. These results suggest that MPC-based dressings may work as an effective support for sustained NT release to reduce DFUs.     

Glucose oxidase incorporated collagen matrices for dermal wound repair in diabetic rat models: a biochemical study

Impaired wound healing in diabetes is a well-documented phenomenon. Emerging data favor the involvement of free radicals in the pathogenesis of diabetic wound healing. We investigated the beneficial role of the sustained release of reactive oxygen species (ROS) in diabetic dermal wound healing. In order to achieve the sustained delivery of ROS in the wound bed, we have incorporated glucose oxidase in the collagen matrix (GOIC), which is applied to the healing diabetic wound. Our in vitro proteolysis studies on incorporated GOIC show increased stability against the proteases in the collagen matrix. In this study, GOIC film and collagen film (CF) are used as dressing material on the wound of streptozotocin-induced diabetic rats. A significant increase in ROS (p?相似文献   

Comparison of the antioxidant properties of wound dressing materials--carboxymethylcellulose,hyaluronan benzyl ester and hyaluronan,towards polymorphonuclear leukocyte-derived reactive oxygen species

In chronic wounds, factors are released which perpetuate inflammatory processes, including polymorphonuclear leukocyte (PMN)-derived reactive oxygen species (ROS), such as superoxide radical (O(2)*-) and hydroxyl radical (*OH) species. The glycosaminoglycan, hyaluronan, has established antioxidant properties towards ROS, although the antioxidant potential of wound dressing biomaterials, such as 75% benzyl esterified hyaluronan (BEHA) and carboxymethylcellulose (CMCH), are less characterised. This study compared the antioxidant properties of high and low molecular weight hyaluronan (HMWT HA and LMWT HA), BEHA and CMCH towards ROS, generated by stimulated PMN in vitro. The antioxidant capacities of each biomaterial were assessed by their inhibition of O(2)*- -induced cytochrome C reduction, generated via PMN stimulation by phorbol myristyl acetate (PMA); and their inhibition of *OH-induced 2-deoxy-D-ribose degradation, generated by PMA stimulated PMN in the presence of a ferric chloride-EDTA chelate. All biomaterials, except LMWT HA, possessed dose-dependent antioxidant properties against O(2)*-, BEHA having greatest antioxidant potential, followed by HMWT HA and CMCH. HMWT HA exhibited the highest dose-dependent antioxidant properties towards *OH, followed by BEHA and CMCH. LMWT HA demonstrated no antioxidant properties towards *OH. These antioxidant activities, particularly towards O(2)*-, may be beneficial in removing the initial source of ROS necessary for the secondary formation of *OH, implicated as a causal factor for the extensive metabolic alterations observed in chronic wounds.     

Attenuation of protease activity in chronic wound fluid with bisphosphonate-functionalised hydrogels

Chronic ulcers are an important and costly medical issue, imposing considerable pain, reduced mobility and decreased quality of life. The common pathology in these chronic wounds is excessive proteolytic activity, resulting in degradation of key factors critical to the ulcer's ability to heal. Matrix metalloproteinases (MMPs), a large family of zinc-dependent endopeptidases, have been shown to have increased activity in chronic wound fluid (CWF), with many authors suggesting that they need to be inhibited for the ulcer to heal. The studies we report here show that the excessive MMP activity in CWF can be inhibited with the bisphosphonate alendronate, in the form of a sodium salt, a functionalised analogue, and tethered to a poly(2-hydroxy methacrylate) (PHEMA) hydrogel. Furthermore, these functionalised alendronate hydrogels appear to be biologically inert as assessed in a three-dimensional ex vivo human skin equivalent model. Together, these results highlight the potential use of a tethered MMP inhibitor to inhibit protease activity in wound fluid. This approach may improve wound healing as it still allows MMPs to remain active in the upper cellular layers of the ulcer bed where they perform vital roles in wound healing; thus may offer an attractive new device-orientated wound therapy.     

不同敷料修复创面的实验研究及临床应用评价

背景：各种材料以多种形式制成的创面敷料近年来发展迅速,复合应用或是制成凝胶、药膜的形式是目前研究和应用的热点。 目的：评价不同敷料在创面的实验研究或者临床应用。 方法：以 “敷料;创面;实验;应用”为关键词,对维普数据库2002-01/2011-10的相关文章进行计算机检索,纳入与敷料和创面相关的随机对照实验类文章,排除重复研究或Meta分析类文章,对22篇文献研究的资料结果进行提取、分析。 结果与结论：纳米银、聚维酮碘、壳聚糖、康复新等是用于实验动物创面的主要材料,分别与中空活性炭纤维、明胶海绵复合或是制成原位凝胶、药膜形式的材料,能最大程度地发挥抗感染、促进创面愈合的作用。壳聚糖、纳米银或者银离子、猪胶原纤维、生物光素、液超妥藻酸盐、痊愈妥等材料可用于临床患者创面,多是以复合或是制成水凝胶等形式,有利于发挥各自性能,更好地促进不同创面愈合。     

甲壳素麻油蜂蜡膏促进糖尿病足创面愈合的研究进展

慢性创面（包括糖尿病足溃疡）合适敷料的应用,创造良好的伤口愈合环境,是临床工作面临的课题。甲壳素、麻油、蜂蜡能有效促进体表创面的愈合,甲壳素麻油蜂蜡膏更能有效促进创面的愈合,为临床敷料的应用提供更多的选择。     

Role of oxidative stress in pancreatic inflammation

Reactive oxygen and reactive nitrogen species (ROS/RNS) have been implicated in the pathogenesis of acute and chronic pancreatitis. Clinical and basic science studies have indicated that ROS/RNS formation processes are intimately linked to the development of the inflammatory disorders. The detrimental effects of highly reactive ROS/RNS are mediated by their direct actions on biomolecules (lipids, proteins, and nucleic acids) and activation of proinflammatory signal cascades, which subsequently lead to activation of immune responses. The present article summarizes the possible sources of ROS/RNS formation and the detailed signaling cascades implicated in the pathogenesis of pancreatic inflammation, as observed in acute and chronic pancreatitis. A therapeutic ROS/RNS-scavenging strategy has been advocated for decades; however, clinical studies examining such approaches have been inconsistent in their results. Emerging evidence indicates that pancreatitis-inducing ROS/RNS generation may be attenuated by targeting ROS/RNS-generating enzymes and upstream mediators.     

“创面床准备”理论实施的系统评价

目的 评价“创面床准备”理论的实施对慢性创面的治疗效果,从而得出一个系统、规范、科学的慢性创面治疗原则.方法 采用Cochrane系统评价方法,检索万方,中国期刊网(CNKI)、中国生物医学文献光盘数据库(CBM)和PubMed等电子数据库,对国内外公开发表的有关“创面床准备”理论实施的研究文献进行荟萃分析.结果 共纳入65个研究、3989例各种慢性创面疾病患者.干预措施包括不同时期选择合适新型敷料、各种生长因子,清创及封闭式负压引流.按测量指标和干预措施进行亚组分析,得出结果:实施“创面床准备”理论,其创面的愈合率[0R=2.12,95％ CI( 1.79,2.52)]、愈合时间[WMD =-12.38,95％ CI(-15.14,-9.62)]、换药次数[WMD=-15.56,95％CI(-20.57,-10.56)]、需要植皮手术的例数[ 0R=0.26,95％ CI (0.10,0.66)]、缩小创面面积[WMD=8.97, 95％ CI( 1.04,16.91)]、植皮手术成功率[0R=5.69,95％CI(2.19,14.82)]均优于对照组,治疗费用[WMD=- 1.53,95％CI(-10.38,7.33)]与对照组差异无统计学意义.结论 实施“创面床准备”理论指导治疗,可以提高创面的愈合率,减少创面愈合时间,加速缩小创面面积,减少换药次数,减少需要植皮手术率,提高植皮手术成功率,并且不增加治疗费用.     

Silver against Pseudomonas aeruginosa biofilms

Silver has been recognized for its antimicrobial properties for centuries. Most studies on the antibacterial efficacy of silver, with particular emphasis on wound healing, have been performed on planktonic bacteria. Our recent studies, however, strongly suggest that colonization of wounds involves bacteria in both the planktonic and biofilm modes of growth. The action of silver on mature in vitro biofilms of Pseudomonas aeruginosa, a primary pathogen of chronic infected wounds, was investigated. The results show that silver is very effective against mature biofilms of P. aeruginosa, but that the silver concentration is important. A concentration of 5-10 mug/mL silver sulfadiazine eradicated the biofilm whereas a lower concentration (1 mug/mL) had no effect. The bactericidal concentration of silver required to eradicate the bacterial biofilm was 10-100 times higher than that used to eradicate planktonic bacteria. These observations strongly indicate that the concentration of silver in currently available wound dressings is much too low for treatment of chronic biofilm wounds. It is suggested that clinicians and manufacturers of the said wound dressings consider whether they are treating wounds primarily colonized either by biofilm-forming or planktonic bacteria.     

Egg white/poly (vinyl alcohol)/MMT nanocomposite hydrogels for wound dressing

Nanocomposite hydrogels on the basis of egg white and poly (vinyl alcohol) (PVA) containing 0, 5, and 10 wt.% of montmorillonite (MMT) nanoclay were prepared by a facile cyclic freezing–thawing technique and their properties investigated for wound dressing application. The morphological, structural, thermal, physical, and in vitro cytotoxic properties of the prepared nanocomposite hydrogel wound dressings (NHWDs) were experimentally studied. The NHWDs had an exfoliated morphology with a porous structure having pores sizes in the nanometric scale. It was shown that MMT acted as cross-linker in the network of NHWDs and improved their thermal stabilities. The prepared wound dressings were transparent and their equilibrium water contents and water vapor transmission rates, as two important factors of wound dressings, were very close to the properties of human skin which means that the prepared wound dressings could interact appropriately with the damaged tissues of wounds and protect them like an artificial skin during the wound healing process. The in vitro cytotoxicity assay also confirmed the non-cytotoxic nature of the prepared NHWDs. It was finally concluded that the prepared egg white/PVA/MMT nanocomposite hydrogels are promising materials to be used as novel wound dressings in wound and burn care.