Photocrosslinkable chitosan hydrogel containing fibroblast growth factor-2 stimulates wound healing in healing-impaired db/db mice

Application of ultraviolet light (UV-) irradiation to a photocrosslinkable chitosan (Az-CH-LA) aqueous solution including fibroblast growth factor-2 (FGF-2) resulted within 30s in an insoluble, flexible hydrogel. About 20% of the FGF-2molecules were released from the FGF-2-incorporated chitosan hydrogel into phosphate buffered saline (PBS) within 1 day, after which no further significant release occurred under in vitro non-degradation conditions of the hydrogel. The FGF-2molecules retained in the chitosan hydrogel remained biologically active, and were released from the chitosan hydrogel upon the in vivo biodegradation of the hydrogel. In order to evaluate its accelerating effect on wound healing, full thickness skin incisions were made on the back of healing-impaired diabetic (db/db) mice and their normal (db/+) littermates. Application of the chitosan hydrogel significantly induced wound contraction and accelerated wound closure in both db/db and db/+ mice. However, the addition of FGF-2 in the chitosan hydrogel further accelerated wound closure in db/db mice, although not in db/+ mice. Histological examination also has demonstrated an advanced granulation tissue formation, capillary formation and epithelialization in wounds treated with FGF-2-incorporated chitosan hydrogels in db/db mice.     

Effects of a collagen matrix containing basic fibroblast growth factor on wound contraction.

We evaluated the effectiveness of basic fibroblast growth factor (bFGF) in inhibiting wound contraction, both alone and in combination with collagen matrix, using a simulated in vivo delayed healing type model. We also studied the mechanisms involved in this inhibition in in vitro experiments using fibroblast populated collagen gels. As a result, we were able to demonstrate that both collagen matrix and bFGF significantly inhibited wound contraction; especially, bFGF acted in a dose-dependent fashion. Interestingly, their combination was much more effective than either collagen matrix or bFGF alone, a finding that was supported by the histopathological data. Wounds treated with collagen matrix, but not control wounds, showed horizontal rearrangement of collagen fibers in dermis as well as evidence of fibroblast proliferation, which was not observed in scar regions surrounded by normal dermis. Using fibroblast-populated collagen gel contraction as an in vitro model, we found that bFGF significantly inhibited contraction. Taking all these results together, it was concluded that collagen matrix is useful not only as a carrier of cytokines such as bFGF, but also for the quick closure of chronic wounds, thereby preventing contracture, which remains one of the most challenging problems in treating this type of wound. Application of bFGF-treated collagen matrix to chronic wounds such as decubitus, and diabetic and leg ulcers may prove to be highly beneficial in clinical practice.     

Nicotine accelerates angiogenesis and wound healing in genetically diabetic mice

Recently, we have discovered an endogenous cholinergic pathway for angiogenesis mediated by endothelial nicotinic acetylcholine receptors (nAChRs). Since angiogenesis plays a major role in wound repair, we hypothesized that activation of nAChRs with nicotine would accelerate wound healing in a murine excisional wound model. In genetically diabetic and control mice full-thickness skin wounds (0.8 cm) were created on the dorsum and topically treated over 7 days with either vehicle (phosphate-buffered saline, PBS) or nicotine (10(-8) mol/L, 10(-9) mol/L; each, n = 5). Wound size was measured over 14 days followed by resection, histological analysis, and quantitation of vascularity. In diabetic animals an agonist (epibatidine, 10(-10) mol/L) or antagonist (hexamethonium, 10(-4) mol/L) of nAChRs as well as the positive control basic fibroblast growth factor (bFGF, 25 microg/kg) were also tested. To further study the role of endothelial nAChRs in angiogenesis, we used an ex vivo vascular explant model. In diabetic mice wound healing was markedly impaired. Nicotine significantly accelerated wound healing as assessed by closure rate and histological score. The effects of nicotine were equal to bFGF and were mimicked by epibatidine and blocked by hexamethonium. Histomorphometry revealed increased neovascularization in animals treated with nicotine. Furthermore, capillary-like sprouting from vascular explants was significantly enhanced by nicotine. In conclusion, agonist-induced stimulation of nAChRs accelerates wound healing in diabetic mice by promoting angiogenesis. We have discovered a cholinergic pathway for angiogenesis that is involved in wound healing, and which is a potential target for therapeutic angiogenesis.     

碱性成纤维细胞生长因子对小鼠脾脏细胞凋亡的抑制作用

目的和方法：Ｘ射线照射结合无血清培养诱导小鼠脾细胞凋亡,采用流式细胞术定量分析碱性成纤维细胞生长因子对脾细胞凋亡的抑制作用,并采用淋巴细胞转化试验「＾３Ｈ」－ＴｄＲ掺入法检测ｂＦＧＦ对脾细胞的增殖效应。结果：小鼠经过不同剂量Ｘ射线照射,脾细胞再经过不同时程无血清培养,都出现了典型了亚二倍体ＡＰ峰,１μｇ＝ｍｌ和２μｇ／ｍＬ     

碱性成纤维细胞生长因子与小儿烧伤创面的愈合

背景：皮肤创面愈合是一个复杂的病理过程,对于创伤和创伤后感染等引起皮肤难愈合的研究一直是临床创面修复的难题,对于碱性成纤维细胞生长因子促进皮肤创面愈合的基础研究相对较多,临床应用研究较少。 目的：对碱性成纤维细胞生长因子促进皮肤创面愈合研究的文献资料趋势进行多层次分析,探讨在小儿烧伤创面愈合中的应用疗效。 方法：以电子检索方式对CNKI数据库学术期刊2002-01/2011-12收录有关碱性成纤维细胞生长因子促进皮肤创面愈合研究的文献进行分析,采用检索词为“碱性成纤维细胞生长因子;创面愈合”,运用数据库的分析功能和Excel软件图表的功能分析数据特征。 结果与结论：CNKI数据库学术期刊2002/2011收录碱性成纤维细胞生长因子促进皮肤创面愈合研究的文献共228篇,文献数量处于平稳发展趋势。《中国组织工程研究与临床康复》杂志收录的文献数量最多为26篇。解放军第304医院产出的文献最多。碱性成纤维细胞生长因子促进皮肤创面愈合研究文献的基金资助项目有16项,基金资助项目的文献共76篇,以国家重点基础研究发展计划(973)项目和国家自然科学基金资助项目的文献最多。碱性成纤维细胞生长因子在小儿烧伤创面愈合中应用的文献虽然较少,但实验结果均显示治疗效果较好,有促进小儿Ⅱ度烧伤创面愈合的作用,而且无不良反应出现。     

Acidic fibroblast growth factor overexpression in corneal epithelial wound healing.

aFGF expression was studied in normal and regenerating cornea of adult rats. aFGF mRNA and proteins were expressed mainly in corneal epithelium but not in stroma. After burning of the epithelium by iodine vapours, the intact epithelial cells migrated to cover the wounded area during the first 4 days and then divided to reconstitute a normal multilayered epithelium 6 days after injury. aFGF mRNA localized by in situ hybridization on regenerating epithelium showed a peak between 6 hr and 2 days after denudation, decreasing to basal levels 6 days later. This induction of aFGF mRNA preceded the increased amount of aFGF peptides, as assessed by indirect immunofluorescence staining. Thus aFGF overexpression is clearly correlated with active migration in epithelial wound healing.     

Monospecific antibodies implicate basic fibroblast growth factor in normal wound repair

Exogenous polypeptide growth factors influence the rate of wound healing and other biological processes, but there is no direct evidence that these peptides have an intrinsic role. To test whether basic fibroblast growth factor is involved in wound repair, rats were implanted with subcutaneous polyvinyl alcohol sponges containing slow-release pellets releasing either a polyclonal neutralizing antiserum directed against basic fibroblast growth factor, preimmune IgG, or nothing. Histological and biochemical evaluation of the granulation tissue that infiltrated the sponges showed anti-basic fibroblast growth factor to cause significant reductions in DNA, protein, and collagen content when compared with either preimmune IgG or placebo at the early stages of wound repair.     

局部注射神经生长因子及碱性成纤维生长因子对大鼠胫骨骨折愈合的影响

目的 探讨局部注射神经生长因子(NGF)及碱性成纤维生长因子(BFGF)对大鼠胫骨骨折愈合的影响及其机制。方法 选取12周龄清洁级成熟雄性SD大鼠 48 只,用数字表法随机分为NGF 组、BFGF组、NGF+BFGF组、对照组4组,各12只;每组大鼠按观察时间点不同随机分为术后2 周及术后4 周2个亚组,每亚组6只。 48 只大鼠均建立左胫骨骨折模型,并行1 mm克氏针髓内内固定。术后第3 天于骨折断端周围局部经皮注射给药:NGF组注射NGF 0.8 μg+0.3 mL生理盐水,BFGF组注射BFGF 1.2 μg+0.3 mL生理盐水,NGF+BFGF组注射NGF 0.8 μg+BFGF 1.2 μg+0.3 mL生理盐水,对照组注射0.3 mL生理盐水;每天注射1次,连续注射7 d。术后2 周、4 周分别行X 线摄片,并采集各组大鼠股动脉血液标本后,采用颈椎离断法处死大鼠。取各组大鼠左胫骨骨痂标本,采用酶联免疫吸附试验(ELISA)检测碱性磷酸酶(ALP)含量;将骨痂组织制成病理切片,行HE染色,观察组织学特点;应用多功能真彩色细胞图象分析管理系统计算切片中骨组织的骨小梁表面成骨细胞指数(IOB)、骨小梁宽度(TW)、骨小梁面积百分比(TV);采用蛋白芯片技术检测大鼠血清中骨形成蛋白-2(BMP-2)、血管内皮生长因子(VEGF)、胰岛素样生长因子-1(IGF-1)的含量。结果 (1)术后2周、4周X线摄片:NGF、BFGF、NGF+BFGF三组大鼠骨折愈合均较对照组快,以NGF+BFGF组愈合最快。(2)ELISA检测:对照组、NGF组、BFGF组、NGF+BFGF组术后2周骨痂组织中ALP的含量分别为(110.02±1.92)、(140.87±2.22)、(136.12±1.23)、(187.44±0.90)ng/mL,术后4周骨痂组织中ALP的含量分别为(91.23±1.47)、(106.62±1.64)、(104.83±1.05)、(130.59±1.18)ng/mL;各观察时点4组间比较,NGF+BFGF组含量最高,组间两两比较差异均有统计学意义(P值均＜0.05)。(3)组织学观察:不同时间点上NGF、BFGF、NGF+BFGF三组大鼠在骨痂的生长、骨小梁形成均较对照组明显,且NGF+BFGF组优于其他组。骨小梁形态学指标测量显示,不同时间点4组间比较,对照组、NGF组、BFGF组、NGF+BFGF组大鼠骨痂内骨小梁IOB、TW、TV均呈上升趋势,NGF+BFGF组最高;组间两两比较,除NGF组与BFGF组各指标差异均无统计学意义(P值均＞0.05),其他指标组间差异均有统计学意义(P值均＜0.05)。(4)蛋白芯片检测:不同时间点4组间比较,除VEGF含量在术后4周时差异无统计学意义(P＞0.05),VEGF术后2周及BMP-2、IGF-1术后2周、4周时,NGF组、BFGF组、NGF+BFGF组均高于对照组,NGF+BFGF组含量最高,组间两两比较,差异均有统计学意义(F=198.285、368.060、2006.017、33.332、292.643,P值均＜0.05)。结论NGF及BFGF均能促进大鼠胫骨骨折愈合,且两者联合应用有协同作用。其机制可能与能够促进骨折愈合过程中VEGF、BMP-2、IGF-1的表达有关。     

Effects of fibroblasts and basic fibroblast growth factor on facilitation of dermal wound healing by type I collagen matrices

Healing of large open dermal wounds is associated with decreased values of the tensile strength even up to 6 months post-wounding. Results of previous studies have shown that healing is facilitated in the presence of a type I collagen sponge by promoting deposition of newly synthesized large-diameter collagen fibers parallel to the fibers of the sponge. In this study healing is evaluated in dermal wounds treated with a collagen sponge seeded with fibroblasts or coated with basic fibroblast growth factor (bFGF). Experimental results indicate that the presence of a collagen sponge results in increased wound tensile strength and increased collagen fiber diameters in the upper dermis 15 days post-wounding in an excisional guinea pig dermal wound model. In comparison, dermal wounds treated with collagen sponges seeded with fibroblasts or coated with bFGF showed increased tensile strengths 15 days postimplantation and increased degree of reepithelialization. These results indicate that fibroblast seeding and bFGF coating in conjunction with a type I collagen sponge matrix facilitate early dermal and epidermal wound healing.     

糖尿病小鼠视网膜碱性成纤维细胞生长因子的表达及细胞损害的电镜观察

目的：为临床研究提供形态学资料。方法：用免疫组化方法，观察碱性成纤维细胞生长因子(bFGF)在糖尿病小鼠视网膜的分布及表达密度；用透射电镜观察不同病程视网膜细胞的损害情况。结果：糖尿病及正常小鼠视网膜各层细胞均表达bFGF，反应强度和密度不同。发病组自6月龄起，居于大血管周围的阳性节细胞密度增加；不同病程bFGF的表达有不同程度的增加。随糖尿病病程延长，细胞超微结构受到不同程度的损害。结论：糖尿病时增多的bFGF直接或间接作用于血管内皮细胞和平滑肌细胞，刺激二者增殖，促进微血管病变的发生；另一方面，减弱视细胞与双极细胞间的信息传递，对糖尿病性盲的发生起重要作用。同时，各种细胞的超微结构及相应的功能也受到不同程度的损害。     

Improved healing of tympanic membrane perforations with basic fibroblast growth factor.

We have investigated the effects of basic fibroblast growth factor (FGF) on the healing of tympanic membrane (TM) perforations. In the first series of experiments, a simple, round 1-mm perforation was made in the membrane and the effects of basic FGF examined. In a second series of experiments, basic FGF was tested on 2-mm perforations in which the borders were folded inward in order to delay normal healing. Topical applications of saline or basic FGF were administered onto gelfoam overlays of the TM perforations in 51 guinea pigs by delivering 5 microliters aliquots of PBS or 5 microliters of PBS containing 1 microgram of basic FGF on the day of surgery and daily thereafter. Repair of the lesions was evaluated 3, 5 or 8 days after surgery. The results show that basic FGF mediates faster healing of TM perforations by inducing rapid proliferation of the subepithelial connective tissue layer.     

Investigation of epidermal growth factor containing liposome formulation effects on burn wound healing

It was aimed to develop liposome formulations containing epidermal growth factor (EGF) and to investigate the healing effects of these formulations on second-degree burn wounds in rats. Multilamellar type liposomes containing EGF were prepared by film formation method. In vitro releases of EGF from liposome formulations were determined using spectrofluorometer. Second-degree standard burn wounds were formed on rats and liposome formulations containing 10 microg/mL EGF (ELP group), EGF solution (ES group), liposome without containing EGF (LP group), and Silverdine ointment (SIL group) were applied daily. Untreated control groups [unburned (S) and untreated (Y) rats] were also evaluated. Biopsies were taken at the 3rd, 7th, and 14th day of the treatment from the wounds and histological observations were performed under transmission electron microscope. Bromodeoxyuridine (BrdU) staining technique was used for immunohistochemical analysis. After trichrome stainings, the thicknesses of the epidermis and the areas of the fibroblast nucleus were measured using a light microscope and Vision Screening Analysis Program. It was observed that the healing in the ELP group was the fastest among all groups (p < 0.05) at the 14th day of therapy. The healing effect in order from highest to lowest efficacy was found to be ELP>SIL>ES groups, respectively, at the 14th day. All results indicated that the EGF-liposome formulation is effective and can be used for the treatment of burn wounds.     

Effect of basic fibroblast growth factor on vascularization in esophagus tissue engineering

We carried out an experimental study to evaluate the effect of basic fibroblast growth factor (bFGF)-containing collagen gel on vascularization in esophageal tissue engineering. We compared an acellular collagen sponge scaffold and an acellular collagen gel scaffold in combination with bFGF using a canine model. The construct was implanted in the cervical esophagus and the regenerated tissue was evaluated one month after surgery. Histological analysis confirmed a significantly large amount of blood vessels in the bFGF-containing collagen gel group as compared to the collagen gel group without bFGF (bFGF (-)). However, in the collagen sponge groups, no difference was observed between the bFGF (+) group and the bFGF (-) group. These results showed that bFGF-containing collagen gel is suitable not only for an acellular scaffold for tissue engineering but also for an effective tropic factor vehicle in vivo.     

Influence of basic fibroblast growth factor on the development of parkinsonian syndrome in mice

Intranasal administration of basic fibroblast growth factor prevented or reduced oligokinesia, muscular rigidity, and weight loss and also prevented mortality in mice with parkinsonism induced by repetitive intraperitoneal injections of 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine. These beneficial effects are probably due to trophic actions of the factor, whereby the degeneration of nigrostriatal neurons is weakened and delayed and their survival is prolonged. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 120, N ^o 9, pp. 260–262, September, 1995     

Placenta growth factor in diabetic wound healing: altered expression and therapeutic potential

Reduced microcirculation and diminished expression of growth factors contribute to wound healing impairment in diabetes. Placenta growth factor (PlGF), an angiogenic mediator promoting pathophysiological neovascularization, is expressed during cutaneous wound healing and improves wound closure by enhancing angiogenesis. By using streptozotocin-induced diabetic mice, we here demonstrate that PlGF induction is strongly reduced in diabetic wounds. Diabetic transgenic mice overexpressing PlGF in the skin displayed accelerated wound closure compared with diabetic wild-type littermates. Moreover, diabetic wound treatment with an adenovirus vector expressing the human PlGF gene (AdCMV.PlGF) significantly accelerated the healing process compared with wounds treated with a control vector. The analysis of treated wounds showed that PlGF gene transfer improved granulation tissue formation, maturation, and vascularization, as well as monocytes/macrophages local recruitment. Platelet-derived growth factor, fibroblast growth factor-2, and vascular endothelial growth factor mRNA levels were increased in AdCMV.PlGF-treated wounds, possibly enhancing PlGF-mediated effects. Finally, PlGF treatment stimulated cultured dermal fibroblast migration, pointing to a direct role of PlGF in accelerating granulation tissue maturation. In conclusion, our data indicate that reduced PlGF expression contributes to impaired wound healing in diabetes and that PlGF gene transfer to diabetic wounds exerts therapeutic activity by promoting different aspects of the repair process.     

Recombinant hepatocyte growth factor accelerates cutaneous wound healing in a diabetic mouse model

We examined effects of recombinant hepatocyte growth factor (HGF) on cutaneous wound healing, using a full-thickness cutaneous excision model in diabetic mice. Topical administration of HGF, as well as basic fibroblast growth factor (bFGF), promoted the rate of wound closure and re-epithelialization. Both HGF and bFGF enhanced expansion of the granulation tissue and stimulated neovascularization on day 7 postwounding, wherein the increase in microvessel density in HGF-treated wounds was higher than that in bFGF-treated wounds. Matrix metalloproteinases (MMP-2 and MMP-9) activities involved in cell migration, angiogenesis, and extracellular matrix (ECM) remodeling, were enhanced by HGF-treatment on day 7. On day 28 postwounding (later stages of wound healing), granulation tissue in bFGF-treated wounds remained to a greater extent than that seen in saline- and HGF-treated wounds. Likewise, bFGF- but not HGF-treatment stimulated DNA synthesis of fibroblasts in granulation tissue, suggesting that HGF stimulates wound healing with lesser degree of susceptibility to cutaneous scarring. We propose that supplement of HGF may be a potential therapeutic approach for treatment of cutaneous ulcer.     

纳米银抗菌水凝胶敷料联合重组人碱性成纤维细胞生长因子在踝部开放骨折创面的应用

背景:纳米银抗菌敷料和水凝胶是新型敷料,用于感染伤口的治疗.重组人碱性成纤维细胞生长因子是临床常应用于开放性伤口的活性多肽类药物.目的:探究外用重组人碱性成纤维细胞生长因子、纳米银抗菌水凝胶敷料在踝部开放骨折清创术后创面愈合中的应用.方法:收集踝部开放骨折清创术后患者99例,根据随机对照表分为对照组、试验A组和试验B组...