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PubMed1989

Granulation tissue in the chick embryo yolk sac blood vessel system.

The development of granulation tissue following acute vascular injury was studied in the yolk sac blood vessel system of chick embryos. Administration of irritant substances was carried out directly on the yolk sac at day 4 of incubation and changes were observed up to 72 h post-application. Reaction nodules developed at the impaired area within 48 h p.a. The nodules were studied by light and electron microscopy and enzyme histochemistry. A proliferation of phagocytosing and mesenchymal cells as well as capillary sprouting took place. Collagenous fibres could be detected within 72 h after administration. The model is suitable for the study of connective tissue proliferation, angiogenesis and possibly drug effects influencing these mechanisms. In accordance with the current legislation for animal protection it is a pain-free test system and can be applied before the onset of reflexes and sensitivity of the chick embryo.

Acid Phosphatase↗
PubMed2006

Is omeprazole or misoprostol superior for improving indomethacin-induced delayed maturation of granulation tissue in rat gastric ulcers?

BACKGROUND AND AIMS: Proton pump inhibitors (PPI) and prostaglandin (PG) preparations are believed to both prevent NSAID-induced gastric ulcers and promote the delayed healing of gastric ulcers by NSAIDs, but it remains unclear which of these drugs is superior. The aim of this study was to clarify which achieved better healing of NSAID-induced gastric ulcers, not only with respect to epithelialization but also repair of the submucosal tissues. METHODS: We used acetic acid to induce gastric ulcers in rats, and compared the changes between a control group, NSAID group, NSAID + PPI group and NSAID + PG group. After removing the stomach of each animal, an ulcer index was calculated and the collagen content and type III collagen content of granulation tissue were measured. We also studied fibroblast dynamics, including proliferation, collagen synthesis, differentiation into myofibroblasts, and apoptosis. RESULTS: Indomethacin prevented re-epithelialization of the ulcers, interfered with fibroblast function, and also delayed the replacement of type III collagen. Omeprazole promoted epithelialization, but could not fully reverse the influence of indomethacin on granulation tissue maturation. A concomitant dose with misoprostolreversed it completely. CONCLUSIONS: From our point of view in this study in the use of experimental ulcers, it was thought that compensation of PG should have priority to gastric acid inhibition in terms of healing of NSAID-induced gastric ulcer.

Analysis of Variance↗
PubMed1992

Multisystem disorder of Punjabi children exhibiting spontaneous dermal and submucosal granulation tissue formation: LOGIC syndrome.

We describe a multisystem disease that affects children of Muslim families originating in the Punjab region of Pakistan and India. An altered cry due to vocal cord thickening, skin ulceration, nail abnormalities, and conjunctival scarring appear in the first few months of life. Progression and spread of the disease in these sites may be accompanied by involvement of other epithelial surfaces. The teeth may exhibit defective enamel formation. Histology reveals the formation of simple granulation tissue arising in the dermis and submucosa which become massively thickened and ulcerated. There is good evidence for an autosomal recessive gene defect, but the actual mechanism of the disease is not known. Medical and surgical therapy have been ineffective in altering the course of this devastating and usually fatal condition. We suggest the term LOGIC (laryngeal and ocular granulation tissue in children from the Indian subcontinent) for this newly established disease.

Child, Preschool↗
PubMedDate not supplied

Squamous cell carcinoma in recessive dystrophic epidermolysis bullosa, presenting as an ulcer that appears to be filled with granulation tissue.

BACKGROUND: Cutaneous squamous cell carcinoma (SCCA) is an important cause of death in patients with the severe form of recessive dystrophic epidermolysis bullosa (RDEB). These cancers often present with features that are different from those of actinically induced SCCA. OBJECTIVE: To emphasize the unusual features with which SCCA can present in RDEB and discuss early recognition of such cancers. METHODS: We present the case of a patient with RDEB who developed a cutaneous ulcer that appeared to be filled with heaped-up granulation tissue. Biopsy, however, showed moderately differentiated squamous cell carcinoma. RESULTS: The patient underwent wide surgical excision of the cancer, with split-thickness skin graft. She has remained disease-free three years later. CONCLUSION: In patients with RDEB, any ulcer that appears to be filled with heaped-up granulation tissue should be biopsied to rule out SCCA.

Biopsy↗
PubMed1994

Topical zinc oxide treatment increases endogenous gene expression of insulin-like growth factor-1 in granulation tissue from porcine wounds.

Application of zinc oxide has been shown to accelerate the healing of both chronic and acute wounds, but the mechanisms are unknown. We quantified the gene expression (mRNA) for one important growth factor, insulin-like growth factor-1 (IGF-1) in 12 full-thickness wounds in each of three domestic pigs treated with or without topical zinc oxide. We used a RNAase protection/solution hybridisation technique to measure IGF-1 mRNA concentrations, which were 50% higher in the granulation tissue in wounds treated with zinc oxide compared with control wounds on days 3-4 (p < 0.05), but not thereafter (up to postoperative day 11). Topical zinc oxide increased the healing rate of wounds compared to the control group (p < 0.01). The cell composition of the granulation tissue was similar in the two groups. The increased gene expression of IGF-1 may be one mechanism by which topical zinc oxide enhances wound healing.

Administration, Topical↗
PubMed1999

A 16-kDa fragment of collagen type XIV is a novel neutrophil chemotactic factor purified from rat granulation tissue.

A neutrophil chemotactic factor has been purified from the homogenate of rat granulation tissues. The purified chemoattractant was a basic protein with heparin-binding site and gave a single band corresponding to a molecular mass of 16 kDa on SDS-PAGE under reducing conditions. The chemoattractant was treated with lysylendopeptidase and the resulting peptides were isolated by reversed-phase HPLC. Amino acid sequences of the peptides were almost identical with the sequence of N-terminal fibronectin type III domain of human collagen type XIV, suggesting that the purified chemoattractant consists mainly of N-terminal fibronectin type III domain and the adjacent heparin-binding site of rat collagen type XIV. The 16-kDa fragment of collagen type XIV dose dependently attracted rat neutrophils and transiently increased the intracellular free Ca2+ concentration of neutrophils. The results suggest that the novel chemoattractant plays a role in neutrophil recruitment in rat inflammation.

Amino Acid Sequence↗
PubMed2002

The CXC chemokine cCAF stimulates precocious deposition of ECM molecules by wound fibroblasts, accelerating development of granulation tissue.

BACKGROUND: During wound repair, fibroblasts orchestrate replacement of the provisional matrix formed during clotting with tenascin, cellular fibronectin and collagen III. These, in turn, are critical for migration of endothelial cells, keratinocytes and additional fibroblasts into the wound site. Fibroblasts are also important in the deposition of collagen I during scar formation. The CXC chemokine chicken Chemotactic and Angiogenic Factor (cCAF), is highly expressed by fibroblasts after wounding and during development of the granulation tissue, especially in areas where extracellular matrix (ECM) is abundant. We hypothesized that cCAF stimulates fibroblasts to produce these matrix molecules. RESULTS: Here we show that this chemokine can stimulate precocious deposition of tenascin, fibronectin and collagen I, but not collagen III. Studies in culture and in vivo show that tenascin stimulation can also be achieved by the N-terminal 15 aas of the protein and occurs at the level of gene expression. In contrast, stimulation of fibronectin and collagen I both require the entire molecule and do not involve changes in gene expression. Fibronectin accumulation appears to be linked to tenascin production, and collagen I to decreased MMP-1 levels. In addition, cCAF is chemotactic for fibroblasts and accelerates their migration. CONCLUSIONS: These previously unknown functions for chemokines suggest that cCAF, the chicken orthologue of human IL-8, enhances healing by rapidly chemoattracting fibroblasts into the wound site and stimulating them to produce ECM molecules, leading to precocious development of granulation tissue. This acceleration of the repair process may have important application to healing of impaired wounds.

Angiogenesis Inducing Agents↗
PubMed1997

Dexamethasone abrogates the fibrogenic effect of transforming growth factor-beta in rat granuloma and granulation tissue fibroblasts.

Administration of TGF-beta, a fibrogenic inflammatory growth factor, promotes fibrosis and scarring. Dexamethasone, an anti-inflammatory steroid, inhibits wound healing and reduces fibrosis. The current studies were initiated to determine whether the co-administration of dexamethasone was able to abrogate the fibrogenic effect of TGF-beta. Polyvinyl alcohol sponges were implanted subcutaneously on the abdominal area of rats and directly injected with vehicle, dexamethasone, TGF-beta, or dexamethasone plus TGF-beta. Dexamethasone was able to block the fibrogenic effect of TGF-beta. Collagen and noncollagen protein synthesis was measured as a function of TGF-beta or dexamethasone concentrations in fibroblasts isolated from granulation tissue. Addition of dexamethasone to cultures treated simultaneously with TGF-beta blocked the fibrogenic response of TGF-beta. To study the molecular regulation of collagen gene expression by TGF-beta or dexamethasone, fibroblasts derived from granulation tissue were stably transfected with the ColCat 3.6 plasmid, which contains the rat pro alpha1(I) collagen promoter linked to the chloramphenicol acetyltransferase (CAT) gene. Dexamethasone decreased CAT activity whereas TGF-beta increased the activity of this reporter gene. The increase in CAT activity observed with TGF-beta treatment was significantly decreased when dexamethasone was added to the cultures, although CAT activity did not return to control level. Since collagen synthesis in fibroblasts treated simultaneously with dexamethasone and TGF-beta1 was found to be the same as that of untreated samples, the data indicate that there is a dexamethasone-mediated posttranscriptional regulation of pro alpha1(I) collagen mRNA. These studies demonstrate that at the in vivo level, the cellular level, and the molecular level, dexamethasone is able to block the fibrogenic effect of TGF-beta.

Animals↗
PubMed1979

Preparation of subcellular fractions from granulation tissue by density gradient centrifugation.

The 7000 g pellet of homogenized mature sponge-induced granulation tissue was fractionated by centrifugation in a stepwise sucrose gradient in order to study the synthesis and secretion of collagen and other components of this tissue on the subcellular level. As indicated by chemical and enzymatic assays, by electron microscopy and by incorporation experiments, the collagen-synthethesizing rough endoplasmic reticulum fraction was isolated free from the secretory vesicles (smooth endoplasmic membranes and Golgi elements) and fibrous extracellular matrix. Collagen differed from other proteins in the distribution among the subcellular fractions. In pulse-chase experiment the translocation of 14C-labelled collagen was demonstrated from the rough endoplasmic reticulum through the secretory vesicles to extracellular fibrillar collagen. This fractionation method will be used to study the modulation of collagen synthesis and secretion in the reparative tissue.

Animals↗
PubMed1992

Heparin induces alpha-smooth muscle actin expression in cultured fibroblasts and in granulation tissue myofibroblasts.

BACKGROUND: Heparin increases alpha-smooth muscle actin expression in smooth muscle cells in vivo and in vitro. It has been recently suggested that alpha-smooth muscle actin expression in fibroblasts is a marker of myofibroblastic differentiation. We have examined the effect of heparin and of four nonanticoagulant heparin derivatives on alpha-smooth muscle actin expression by fibroblasts in vitro and in vivo. EXPERIMENTAL DESIGN: For in vitro experiments, heparin was added for 7 days to different fibroblastic cultures. We studied cell proliferation and alpha-smooth muscle actin protein and mRNA expression. For in vivo studies, osmotic minipumps filled with NaCl or tumor necrosis factor-alpha without or with nonanticoagulant heparin were implanted subcutaneously. After 14 days, newly accumulated connective tissues around the pumps were processed for immunofluorescence and electron microscopic and biochemical studies. RESULTS: In vitro, heparin inhibited proliferation and increased the expression of alpha-smooth muscle actin protein and mRNA. Analysis of [3H]thymidine incorporation in synchronized cells suggested that heparin produces a selection of alpha-smooth muscle actin expressing cells. In vivo, the local application of tumor necrosis factor-alpha resulted in formation of a typical granulation tissue: immunofluorescence showed that accumulated fibroblastic cells express alpha-smooth muscle actin only in the presence of heparin derivatives. In tumor necrosis factor-alpha treated animals, electron microscopic examination established the presence of myofibroblasts, but alpha-smooth muscle actin was expressed in microfilament bundles only in the presence of heparin derivatives. CONCLUSIONS: These results show that heparin and its nonanticoagulant derivatives influence the expression of alpha-smooth muscle actin in fibroblastic cells both in vitro and in vivo and that this effect is probably related to the selection of a particular cell subpopulation. They suggest a possible role for heparin during the formation and evolution of granulation tissue.

Actins↗
PubMed1992

Interstitial and airspace granulation tissue reactions in lung transplant recipients.

Twenty-three transbronchial and open-lung biopsies from patients who had received a lung allograft displayed fibromyxoid plugs of granulation tissue within airways, airspaces, and the interstitium in a patchy distribution. This granulation tissue-like reaction was identified in three clinicopathologic settings. First, 11 cases occurred with acute lung rejection, of which four cases had been partially treated with steroids for a previous rejection episode. Second, in seven cases the fibromyxoid tissue represented the healing phase of previously diagnosed diffuse alveolar damage resulting from preservation (harvest) injury to the allograft. Third, five cases were related to infection: herpes, Pseudomonas, Serratia, Staphylococcus, and Pneumocystis pneumonias. Although organizing pneumonia-like responses usually suggest an infectious episode, this reaction may be seen as a manifestation of acute lung rejection or ischemic lung injury.

Biopsy↗
PubMed1990

Collagen distribution in developing experimentally induced granulation tissue. A morphometric study.

Viscose cellulose sponges were implanted subcutaneously on the back of full-grown Sprague-Dawley rats. Seven, 14, 21, 28, 42, 60 and 90 days after implantation, groups of 12 animals decapitated and the sponges were removed and processed for light microscopy. Five microns sections were stained with Picro-Sirius Red. Morphometry was performed on the zone of ingrowth and the collagen. The intersectional variation in the morphometrically determined collagen density within the sponges was below 20%. The hydroxyproline content was determined biochemically in 5 microns sections of sponges implanted for 14, 42, 60 and 90 days. A positive correlation (rho = 0.79, p less than 0.0001) was observed between the biochemically and morphometrically determined collagen contents. The morphometric determinations showed a steady increase in the granulation tissue ingrowth. At day 60 the ingrowth was complete. There was an increasing collagen density from days 7 and 14 through days 21 and 28, followed by a nearly steady state up to day 90 and a significantly higher collagen density peripherally than centrally in the day 42 sponges. The study has shown that morphometric collagen determination at light microscopical level using Sirius Red-stained sections may add quantitative data describing the dynamic changes in collagen content and distribution within developing granulation tissue.

Animals↗
PubMed1983

Can heterologous collagen enhance the granulation tissue growth? An experimental study.

A purified and lyophilized sponge of native bovine tendon collagen (Gelfix) was used to plug gastric, jejunal and colonic surgical holes (phi = 6 mm) on 78 white rats. In a control group (18 rats) colonic holes were plugged with autologous epiploon. The mortality rate due to generalized peritonitis, was higher (Fischer test = 4%) in the control group than in animals treated with the heterologous collagen. The healing process was evaluated by light microscopy at sequential intervals ranging from 4 to 120 days after implantation. The heterologous collagen biological activity was evaluated as follows: 1. The sponge was labelled with Tyrosine 125I or mixed with Collagen Methyl 14C methylated. Then the radioactivity of the isolated granulation tissue cells was evaluated. The electrophoresis of these lysed cells, as well as histoautoradiography were performed. 2. Indirect immunofluorescence studies employing rabbit antibodies against fibronectin were also carried out. The heterologous collagen seemed to act as a mechanical support and as a stimulus to chemotactic migration of fibroblasts. It also appeared to participate as a nutrient substitute in the metabolic activity of the granulation tissue. Finally, heterologous collagen fibers were found to be massively bound by fibronectin.

Animals↗
PubMed1996

Acceleration of granulation tissue ingrowth by hyaluronic acid in artificial skin.

Hyaluronic acid (HA), which is known to play an important role in wound healing, was incorporated in an artificial skin material and studied for its potential to create a wound bed which would support a skin graft. Collagen sponge based artificial skin was soaked in 0.3% HA in phosphate buffered saline and grafted onto skin defects in rats. Control grafts were soaked in normal saline solution. HA incorporated implants and control implants were simultaneously grafted onto wounds made on either side of the spine. To examine the effect of HA incorporation, the percentage area of cellular tuft infiltration and the number of capillaries present in the graft matrix were evaluated at 7 and 14 days after the operation. At postoperative day 7, there was a statistically significant difference in the number of capillaries in the matrix of the experimental versus the control implants. There was no difference in the percentage area of cellular tuft infiltration. At postoperative day 14, all implants exhibited better ingrowth of granulation tissue than at day 7. The differences between the experimental and control implants were statistically significant with respect to both the percentage area of cellular tuft infiltration and the number of capillaries. It is therefore concluded that in artificial skin HA incorporation accelerates the ingrowth of granulation tissue, making a more suitable graft bed.

Animals↗
PubMed1975

Granulation tissue formation in oophorectomized rats treated with female sex hormones. I. A histological study.

Granulation tissue formation was studied in experimental granulomas up to three weeks in oophorectomized rats treated with female sex hormones. The influence of estrogen and progesterone during the early phase of healing could sucessfully be followed histologically. The amount of DNA and hemoglobin in the granulomas correlated well with the amount of cells and the capillary bed found in the histological sections.

Animals↗
PubMed2004

Wound splinting modulates granulation tissue proliferation.

Attachment of the extracellular matrix to a substratum is important for fibroblast survival and proliferation in three-dimensional in vitro culture systems. We hypothesized that wound matrix attachment in a wound splinting model would modulate wound cell proliferation in vivo. Male rats were excisionally wounded on the dorsum, and a splint was sutured to the wound edge. In one experiment (N = 12), 6 rats were desplinted on day 5, and then all were sacrificed 24 h later, 6 h after 5-bromo-2'-deoxyuridine (BrdU) injection. In the second experiment (N = 18), 6 rats each were desplinted, desplinted with wound edge release, or not disturbed, followed by BrdU injection and sacrifice 24 h later. BrdU-labeled nuclei were quantified on frozen sections of granulation tissue, cut at three different levels. In the first experiment, the percentage of BrdU-positive nuclei per high power field (hpf) in the splinted vs. desplinted animals was 6.15 +/- 2.45 (S.D.) vs. 3.03 +/- 1.58%* p<0.001, ANOVA. In the second experiment, the number of BrdU-positive per hpf was 33.1 +/- 17.4 vs. 14.5 +/- 17.1 vs. 10.2 +/- 9.1* (splinted vs. desplinted vs. desplinted/released); *p<0.001 [analysis of variance (ANOVA)]. Removal of the wound splint decreased the rate of BrdU-labeled cells in the granulation tissue by approximately 50%; complete disruption of wound matrix attachment may have decreased this rate even further. Wound cell proliferation is modulated by lateral attachment of the wound matrix.

Analysis of Variance↗
PubMed1996

Keratinocyte growth factor induces granulation tissue in ischemic dermal wounds. Importance of epithelial-mesenchymal cell interactions.

BACKGROUND: Keratinocyte growth factor acts specifically on epithelial cells and is presumed to play an important role in tissue repair. OBJECTIVE: To examine the wound-healing effects of keratinocyte growth factor under hypoxic conditions in vivo and in vitro. DESIGN AND INTERVENTIONS: Dermal ulcers were created in the ischemic ears of 40 anesthetized young female rabbits. Either recombinant keratinocyte growth factor (rKGF) or buffer was applied to each wound. Wounds were bisected and analyzed histologically at days 7 and 10 after wounds were created. For the in vitro study, normal keratinocytes were treated with rKGF (20 ng/mL) and cultured under hypoxic (3.5% oxygen) conditions. The conditioned media were collected at 48 and 72 hours. MAIN OUTCOME MEASUREMENTS: The amount of epithelial growth and deposition of granulation tissue were measured in all wounds. The amount of transforming growth factor alpha in keratinocyte-conditioned media was measured by using a sensitive radioimmunoassay. A proliferation assay of dermal fibroblasts, treated with conditioned media, was also performed under 3.5% oxygen culture conditions. RESULTS: The rKGF (range, 5-40 micrograms per wound) that was applied significantly increased new epithelium by greater than 70% (P = .03) at days 7 and 10 after wounds were created. A significant increase in new granulation tissue formation (170%) was also observed in rKGF-treated wounds at day 10, at a dose of 40 micrograms per wound (P < .002). The amount of transforming growth factor alpha protein in the conditioned media that were treated with rKGF (20 ng/mL) increased by 26.8% and 171% at 48 and 72 hours, respectively, over that of controls. The conditioned media from rKGF-treated keratinocytes, grown for 72 hours, resulted in a 51% increase in the proliferation of primary rabbit dermal fibroblasts. CONCLUSION: Keratinocyte growth factor enhances the wound-healing process of ischemic ulcers, indicating that epithelial-mesenchymal cell interactions are critical for the healing of wounds under ischemic conditions and possibly under normal conditions as well.

Animals↗
PubMed1977

Influence of somatostatin on peripheral leucocyte count and granulation tissue in man and rats.

Subcutaneous injection of synthetic protamin-zink-somatostatin completely prevents endotoxin-induced leucocytosis in normal rats. Piromen-induced elevated stab neutrophil, neutrophil and monocyte counts remain within the normal range during somatostatin administration. There is an inhibiting effect of synthetic protamin-zink-somatostatin on the wet weights of granulation tissue of cotton pellet granulomata, too. Incorporation of 35S-sulfate in sulfated mucopolysaccharides of granulation tissue in cotton pellet granulomata is not inhibited. Intravenous administration of synthetic linear somatostatin decreases stab neutrophil and neutrophil blood count in patients with acute bacterial leucocytosis. After the termination of somatostatin infusion a rebound phenomenon occurs. In healthy subjects lymphocyte count increases during somatostatin infusion. This effect can not be demonstrated in patients with bacterial leucocytosis.

Adult↗