Effects of basic fibroblast growth factor administration on vascular changes in wound healing of rat palates.

OBJECTIVE: The purpose of this study was to investigate the vascular changes induced by mucoperiosteal denudation of rat palate and to elucidate the effects of basic fibroblast growth factor (bFGF) administration on the palatal vascular network in wound healing. METHODS: A total of 117 male Wistar rats were used for the study on their 20th postnatal day. The animals were divided into three groups: a scar formation group, a basic fibroblast growth factor group, and a control group. The scar formation and basic fibroblast growth factor groups had lateral mucoperiosteum excised from the palate. In the basic fibroblast growth factor group, a solution of basic fibroblast growth factor was injected into the operated area 1 week after excision. At 6, 8, and 10 weeks postoperatively, palatal vascular changes were investigated by immunohistochemical staining and corrosion cast techniques. RESULTS: Throughout the experimental period, there were significantly fewer vessels in the scar formation group than in the control and basic fibroblast growth factor groups. In the basic fibroblast growth factor group, the elongation of new vessels and capillary proliferation proceeded, and after 10 weeks a highly organized vascular network was established. The scar formation group showed few Volkmann's canals that were shrunken or closed, whereas the basic fibroblast growth factor group evidenced Volkmann's canals with arterioles or venules, as seen in the control. CONCLUSIONS: The results suggested that injection of basic fibroblast growth factor into palatal wounds improves the vascular supply to the operated mucosa and underlying bone during and after palatal wound healing, which may contribute to tissue remodeling of the palate during growth.     

Human deciduous teeth dental pulp cells with basic fibroblast growth factor enhance wound healing of skin defect

In this research, we examined the effect on wound healing applying basic fibroblast growth factor (b-FGF) that is approved for clinical use to enhance wound healing and human deciduous teeth dental pulp cells (hDPCs) in clinics, but that have been attracting attention as a novel stem cell source in recent years. Human deciduous teeth were harvested from healthy volunteers, and hDPCs were isolated. We used a nude mouse full-thickness skin defect model and evaluated wound healing by macroscopic view and histologic and histomorphometric analysis. The mice were randomly divided into 4 groups: phosphate-buffered saline-treated group (control group), b-FGF-treated group (b-FGF group), hDPC-treated group (hDPC group), and hDPC and b-FGF-treated group (hDPC/b-FGF group). Basic fibroblast growth factor and hDPC groups accelerated wound healing compared with the control group. There was no statistically significant difference in wound healing observed between the hDPC and b-FGF groups. The hDPC/b-FGF group demonstrated accelerated wound healing compared with other groups. At day 14, PKH26-positive cells were surrounded by human type I collagen in hDPC and hDPC/b-FGF groups in immunohistologic evaluation. Significantly increased collagen fibril areas in wound tissues were observed in b-FGF, hDPC, and hDPC/b-FGF groups as compared with the control group at days 7 and 14. Our results showed that the hDPC/b-FGF group significantly promotes wound healing compared with other groups. This study implies that deciduous teeth that are currently considered as medical spare parts might offer a unique stem cell resource for potential of new cell therapies for wound healing in combination with b-FGF.     

Low-intensity pulsed ultrasound enhances palatal mucosa wound healing in rats

PurposeLow-intensity pulsed ultrasound (LIPUS) has been used in fracture treatment to shorten the time needed for biological wound healing. However, the influence of LIPUS exposure on oral wound healing has not been sufficiently investigated. This study was conducted to evaluate low-intensity pulsed ultrasound on wound healing in palatal excisional wounds of rats.MethodsExcisional wounds, 5 mm in diameter, were made in the center of the palate of rats. Animals were divided into four experimental and control groups (1-week after LIPUS exposure, 1-week control, 2-week after LIPUS exposure, and 2-week control). The affected area in the experimental group was exposed to LIPUS, daily frequency: 3 MHz, intensity: 160 mW, exposure time: 15 min. Specimens were fixed in 10% neutral formalin solution immediately after sacrifice. The wound was measured histologically.ResultsWound width in the LIPUS group tended to be smaller than that of the control group. The experimental group in both 1-week and 2-week groups showed that unhealed areas were significantly smaller by LIPUS than those in the control groups (P < 0.05).ConclusionOur results suggest that the use of LIPUS on palatal excisional wounds was effective in promoting epithelial and connective tissue closure.     

Quantitative evaluation of myofibroblast apoptosis during wound healing in rat palate after post-operative administration of basic fibroblast growth factor (bFGF)

Abstract

Objective. Excessive wound contraction apparently inhibits maxillary growth; thus, myofibroblast apoptosis needs to be accelerated in mucoperiosteal denudation after palatoplasty. The aim of this study was to evaluate myofibroblast apoptosis during wound healing in mucoperiosteal denudation of rat palates immediately after post-operative administration of basic fibroblast growth factor (bFGF). Materials and methods. A total of 100 male Wistar rats aged 20 days were divided into control, scar, sham and bFGF groups (n = 25 each). In the scar, sham and bFGF groups, mucoperiosteum was removed from the palate and fibrin glue was applied to the exposed bone surface immediately after surgery. In the bFGF group, 10 μL of 2 μg/μL bFGF solution was injected into the operated area beneath the fibrin glue. At 2, 5, 7, 14 and 28 days post-operatively, myofibroblast apoptosis during the wound healing process was investigated by double immunofluorescence staining. The apoptotic area of myofibroblasts was measured using image software. Results. In the bFGF group, at 2 days, apoptosis of myofibroblasts in the lamina propria and submucosa was marked, as compared with the other three groups and apoptosis of myofibroblasts was scarcely seen at 5 days. At 5 and 7 days, the apoptotic area of myofibroblasts in the bFGF group was statistically significantly smaller when compared to the scar and sham groups. Conclusion. The results confirmed that bFGF injection immediately after surgery accelerated apoptosis of myofibroblasts in mucoperiosteal denudation of rats. This may reduce maxillary growth retardation due to excessive wound contraction.     

Basic fibroblast growth factor and epidermal growth factor reverse impaired ulcer healing of the rabbit oral mucosa

BACKGROUND: The therapies for refractory ulcers on the oral mucosa are symptomatic and very unsatisfactory. We hypothesized that application of growth factors might be able to achieve successful remission of the lesion. We evaluated the effects of systemic administration and topical application of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on impaired wound healing of ulcers in the rabbit gingiva. METHODS: Almost uniform round ulcers could be created on the gingiva of the rabbits by chemical injury with acetic acid. When the submandibular glands were removed or i.v. injection of cisplatin (CDDP) and peplomycin sulfate was performed before ulcer formation, healing of the ulcers took longer than in untreated rabbits. To ascertain whether or not human EGF and bFGF affect rabbit cells, we first examined the effects of EGF and bFGF on the proliferation of the cells derived from rabbit gingiva. We then applied EGF or bFGF in these impaired healing models. RESULTS: EGF and bFGF promoted proliferation of the fibroblasts, and EGF also promoted proliferation of the keratinocytes isolated from gingival tissue of rabbits in vitro. Systemic injections of EGF and bFGF in rabbits, which had their submandibular glands removed, and topical application of bFGF accelerated healing of ulcers created in rabbits injected with CDDP and peplomycin sulfate. The ability of bFGF to promote the healing of ulcers was much greater than that of EGF. CONCLUSION: Basic FGF may be effective for refractory oral mucosal lesions.     

应用数码显微镜评价重组牛碱性成纤维细胞生长因子辅助治疗萎缩性舌炎的临床效果

目的：应用数码显微镜观察重组牛碱性成纤维细胞生长因子（recombinantbovinebasicfibroblastgrowthfactor,rb-bFGF）辅助治疗萎缩性舌炎的疗效。方法：39例维生素B12缺乏的萎缩性舌炎患者,随机进入治疗组（21例）和对照组（18例）。采用数码显微镜,检测患者初次就诊及第7、14d复诊时舌乳头形态,同时记录舌痛、味觉改变的程度。比较两组患者在初诊及两次复诊时上述3项指标的改善情况。结果：第7d复诊,治疗组患者总体舌黏膜萎缩的改善程度高于对照组（P〈0．05）,其舌痛存在时间短于对照组护（P〈0．05）;第14d复诊,两组治疗效果趋于一致。结论：rbFGF能够减轻炎症反应、促进舌乳头生长并缩短疼痛时间。     

Ultrastructure of squamous epithelium during wound healing in palatal mucosa of guinea pigs

The ultrastructure of the epithelium of wounded guinea pig palate was studied. Twenty-four biopsies (animals) were used. Biopsies were taken 18, 48, 96, and 120 h after wounding. Essential changes in epithelial cell structure in response to wounding were as follows: microfilaments, suggested to represent actin-containing filaments, became evident in the cortical cytoplasmic zone. Micro tubules prevailed in the peripheral cytoplasm. Tonofilament bundles assumed the character of short tufts or delicate bundles and. the amount of non-bundled tonofilaments was increased. These observations suggest changes in the cytoskeletal/contractile system facilitating cell motility. The structure of lysosomal profiles indicates an enhanced phagocytic capability. Influence of fixation on preservation of non-membrane-bound organelles is suggested based on comparable morphometric data.     

碱性成纤维细胞生长因子对体外大鼠牙胚分化的影响

目的：观察外源性ｂＦＧＦ对体外培养牙胚发育和分化的作用。方法：采用体外培养１７ｄＳＤ胎鼠第一磨牙牙胚，培养液中加入１０ｎｇ／ｍｌ人重组ｂＦＧＦ（ｈｂＦＧＦ），分别培养３、６、９ｄ，组织学观察。结果：外源性ｈｂＦＧＦ可使大鼠牙胚的细胞分化和牙本质基质分泌加快。结论：外源性ｂＦＧＦ可以促进大鼠牙胚的发育，加速牙齿发育。     

Cell junctions in squamous epithelium during wound healing in palatal mucosa of guinea pigs

The nature and distribution of cell junctions of the epithelium of wounded guinea pig palate were .studied. Biopsies, 32 in total, were taken 13, 48, 96, and 120 h after wounding. Desmosomes were present in the wound epithelium throughout the observation period. Primitive desmosomes, indicating de novo formation of desmosomes, were observed at the front of advanced migratory lips (48 h) and in the central region and the wound epithelium after bridging (96 and 120 h). Similarly, de novo formed hemidesmosomes were seen. They occurred along epithelial cell membranes exposed to the fibrinous wound exudate during the migratory lip stages (18 and 48 h) and after epithelial bridging. The observations .suggest that wounding induced changes in the mechanical properties of the tissue. Gap junctions were present in migrating epithelium preferably at the stem of the lips and in the epithelium after bridging, which suggests their participation in regulation of the epithelial cell differentiation. Tight junctions, capable of acting as seals, were absent or fragmentary at best. A relative decrease in the volume density of intercellular spaces in migrating epithelium as compared to intact epithelium, suggests an in vivo tissue reaction.     

Acceleration of rat salivary gland tissue repair by basic fibroblast growth factor

A model of atrophic rat submandibular gland was used to examine the ability of basic fibroblast growth factor (bFGF) to accelerate tissue repair. The gland duct was separated carefully from associated blood vessels and nerve, and ligated with a 8-0 suture under a surgical microscope. Two weeks after ligation, the glandular tissue showed severe atrophy and weight loss (to 26% of that in a sham-operated group). Thereafter, the ligature was removed and various amounts of bFGF, isoproterenol or saline were instilled retrogradely through the duct. Both isoproterenol and bFGF increased cell proliferation significantly. bFGF accelerated the proliferation of various cell types, including both acinar and ductal. The proliferative effects of bFGF peaked at a dose of 1 ng/gland. When bFGF (1 ng/gland) was administered to the atrophic gland, its weight increased to 125% of the glands in saline-treated control animals after 2 weeks. The effects of bFGF were also examined in normal submandibular glands: bFGF stimulated cell proliferation, but the effective concentration was at least 50 times higher than that required in the atrophic gland. The results from immunohistochemical tests against anti-FGF receptor-type 1 antibody demonstrated increased immunoreactivity in the damaged gland, which might be involved in the difference in the response to bFGF between damaged and normal glands. Overall, the results indicate that bFGF can accelerate tissue repair in salivary gland.     

过量氟对大鼠切牙碱性成纤维细胞生长因子表达的影响

目的 研究过量氟对大鼠切牙碱性成纤维细胞生长因子（basic fibroblast growth factor，bFGF）表达的影响，从分子水平探讨氟牙症的发病机制。方法将20只Wistar大鼠按随机数字表法随机分为两组：对照组（饮用蒸馏水）和实验组（饮用100mg／L氟化水），复制氟牙症动物模型，8周末处死动物，获取切牙标本，免疫组化染色观察bFGF在大鼠切牙的表达定位及其在对照组与实验组切牙表达的变化。结果bFGF在分泌期成釉细胞、成牙本质细胞中均呈阳性表达。实验组bFGF的表达明显弱于对照组，差异有统计学意义（P〈0．01）。结论过量氟可抑制bFGF的表达，从而影响上皮与间充质之间的相互介导作用，导致釉质发育障碍。     

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目的 探讨碱性成纤维细胞生长因子(bFGF)基因修饰的组织工程化复合物对牙龈组织缺损再生的影响.方法 本研究于2007年7月至2009年1月在福建省高校口腔医学重点实验室进行.利用bFGF基因转染Beagle 犬牙龈成纤维细胞(GFs),并将其接种于脱细胞真皮基质(ADM)形成组织工程化复合物,植入Beagle犬的人工牙周组织缺损区,通过对治疗前后牙周指标测量分析,评价该组织工程化复合物对牙龈组织再生的影响.结果 实验前后附着丧失差值测定显示GFs与ADM复合物组及转染bFGF基因的GFs复合物组之间无显著性差异,与空白对照组相比均有明显增加(P＜ 0.05).角化龈宽度差值,6周时转染基因组有明显增加,12周时两实验组均比空白对照组明显增加(P＜0.05).结论 复合GFs的ADM可能有助于改善附着丧失的程度及角化龈宽度,而bFGF的作用并不肯定.组织工程技术能有效促进牙龈组织再生.     

碱性成纤维细胞生长因子对兔下颌骨骨折愈合中细胞增殖的影响

目的：研究重组人碱性成纤维细胞生长因子（recombinant human basic fibroblast growth factor,rhbFGF）对兔下颌骨骨折愈合中细胞增殖的影响。方法：在兔下颌骨骨折动物模型中,以牛胶原蛋白I型为载体,餐源性应用rhbFGF于骨折局部。分别于术后1、2、4、8、12周时取材,采用ABC方法进行增殖细胞核抗原（proliferating cell nuclear antigen,PCNA)免疫组化染色。结果：术后、2周时rhbFGF治疗组骨痂内PCNA阳性细胞比率明显高于对照组（P＜0．01）。结论:rhbFGF在兔下颌骨骨折愈合早期,对细胞的增殖起十分重要的促进作用。     

Effects of basic fibroblast growth factor on cultured rat and human submandibular salivary gland cells

Basic fibroblast growth factor (bFGF) is a strong mitogen for most mesoderm- and ectoderm-derived cells. Although bFGF exists in rat and human salivary glands, its physiological role in those glands is unknown. In this study, the effects of bFGF were investigated in monolayer culture of normal rat and human submandibular gland cells. Epithelial cells from rat and human submandibular glands were cultivated with the aid of 3T3 cells as a feeder layer. The effects of different concentrations of bFGF on the second passage of these cultured cells were examined. In both the rat and human cells, the percentage of bromodeoxyuridine (BrdU)-positive cells gradually increased up to 50 ng/ml, and then increased sharply at 100 ng/ml. However, at concentrations higher than 100 ng/ml, the percentages of BrdU-positive cells reached a plateau. In both rat and human cells, total cell numbers at 100 ng/ml bFGF were significantly higher than those of the control group from culture day 4. On the other hand, the morphology of the cultured cells showed no difference either with or without bFGF. These results indicate that a major effect of bFGF on salivary gland epithelial cells is to act as a mitogenic stimulus.     

碱性成纤维细胞生长因子对大鼠下颌骨缺损Bio-oss植骨的影响

目的研究外源性碱性成纤维细胞生长因子对大鼠下颌骨缺损Bio-oss植骨愈合的影响。方法选用48只体重约450g的Wistar大鼠随机分为2组,实验组24只,对照组24只。在大鼠左侧下颌角处制备5mm×4mm×2mm的骨缺损区。实验组植入含有基因重组大鼠碱性成纤维细胞生长因子的Bio-oss颗粒。对照组植入含生理盐水的Bio-oss颗粒。于术后2、4、6、8周分四批处死大鼠,取其左侧下颌骨进行X线和组织学检查。结果术后各个阶段实验组骨愈合的速度均高于对照组。结论外源性碱性成纤维细胞生长因子与Bio-oss复合后对大鼠下颌骨缺损的修复效果明显优于单纯应用Bio-oss骨。     

Regeneration of periodontal tissues by basic fibroblast growth factor

Several growth factors (or cytokines) have recently received attention because of their ability to actively regulate various cellular functions of periodontal ligament (PDL) cells and the effects of topical application of such factor(s) on periodontal tissue regeneration has been evaluated. In this study, we examined the role of basic fibroblast growth factor (bFGF) in the wound healing and regeneration of periodontal tissues. Alveolar bone defects (such as 2-wall, 3-wall and furcation class II bone defects) were created surgically in beagle dogs and primates. Recombinant bFGF was topically applied to the artificial bony defects. Six or 8 wk after application, the periodontal regeneration was morphologically and histomorphometrically analyzed. In all sites where bFGF was applied, significant periodontal ligament formation with new cementum deposits and new bone formation was observed in amounts greater than in the control sites. We found it noteworthy that no instances of epithelial down growth, ankylosis or root resorption were observed in the bFGF sites. In vitro studies demonstrated that bFGF enhances the proliferative responses of human PDL cells, which express FGF receptor-1 and -2, but inhibits the induction of alkaline phosphatase activity and mineralized nodule formation by PDL cells. Interestingly, we observed that the mRNA level of laminin in PDL cells, which plays an important role in angiogenesis, was specifically upregulated by bFGF stimulation, but that of type I collagen was downregulated. The present study demonstrates that bFGF can be applied as one of the therapeutic modalities which actively induce periodontal tissue regeneration. The results of in vitro studies suggest that by suppressing the cytodifferentiation of PDL cells into mineralized tissue forming cells, bFGF may play important roles in wound healing by promoting angiogenesis and inducing the growth of immature PDL cells, and may in turn accelerate periodontal regeneration.     

The effect of basic fibroblast growth factor on delayed-replanted monkey teeth

BACKGROUND: Basic fibroblast growth factor (bFGF; FGF-2) has been reported to facilitate wound healing and periodontal regeneration in experimental alveolar bone defects. The purpose of this study was to evaluate histologically the effect of topically applied bFGF with or without fibrin glue on delayed-replanted monkey teeth prone to replacement resorption. METHODS: Forty-five roots from five monkeys were endodontically treated aseptically and then extracted as atraumatically as possible. Ten negative control roots were replanted immediately, while 12 positive control roots were allowed to bench dry for 1 hour prior to replantation, both without further treatment. Roots in the two experimental groups were bench dried for 1 hour, rinsed with saline, and then replanted into sockets filled with bFGF with (11 roots) or without (12 roots) fibrin glue. After 12 weeks, histological sections were prepared and evaluated according to morphometric analysis as complete healing or unfavorable healing composed of inflammatory resorption and replacement resorption. RESULTS: Kruskal-Wallis and Mann-Whitney U tests showed teeth in the negative control group to have significantly higher complete healing (98.88% +/- 2.30%) and significantly lower unfavorable healing (1.12% +/- 2.30%) than the positive control group and the experimental groups. bFGF/fibrin glue group showed higher occurrence of complete healing (39.06% +/- 41.62%) compared to the bFGF group (25.28% +/- 28.85%) and the positive control group (16.58% +/- 19.60%), although the differences were not significant. Comparing the complete and unfavorable healing, there was no significant difference in the bFGF/fibrin glue group (P = 0.47), but the differences were significant in the other groups (P < 0.05). CONCLUSION: Topical application of bFGF with fibrin glue showed an insignificantly higher occurrence of complete healing in delayed-replanted monkey teeth.