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PubMedDate not supplied

Different metabolism of hexose sugars and sucrose in wound fluid and in fibroblast cultures derived from granulation tissue, hypertrophic scar and keloid.

Viscose cellulose sponge implants in the rat and fibroblast cultures established from granulation tissue, hypertrophic scar, or keloid were treated with different concentrations of glucose, fructose, galactose, mannose, and sucrose. The concentrations of the above-mentioned sugars in wound fluid and cell culture medium were examined at the termination of experiments by liquid chromatography. Results showed that glucose was present in wound fluid in relatively low levels. In addition to glucose, only mannose was found in wound fluid. On the other hand, cell culture studies showed that virtually all the added sugars were found in cell culture medium. The most prominent exception was the decreased concentration of mannose in keloid fibroblast cultures. In addition, glucose concentration in culture medium of keloid fibroblasts was constantly very low except in mannose-treated cultures where the consumption of glucose was dose-dependently decreased compared to increased mannose concentration. Similarly, increased concentrations of galactose and mannose resulted in dose-dependent lowered consumption of glucose in granulation tissue and hypertrophic scar fibroblasts. These findings suggest that the sugar metabolism may differ in various fibroblast cultures. Further, at least in our wound model, only glucose and mannose are present in wound fluid, and excess sugar is rapidly cleared from wound fluid.

Animals↗
PubMed1975

Protein synthesis by granulation tissue in bipedicle tube flaps.

The protein synthesis and the collagen synthesis by 2- to 7-week-old granulation tissue from implanted cylinders in bipedicle tube flaps on rabbits were measured in vitro by the incorporation of 14C-proline in proteins and in collagen hydroxyproline. Granulation tissue from cylinders implanted in skin folds were used as controls. In the skin flap group the protein synthesis was high at week 2, after which it decreased to week 5. In the controls the protein synthesis was the same as in the skin flap group at week 2. A decrease occurred between weeks 2 and 4 and a further decrease between weeks 6 and 7. At weeks 5 and 6 the protein synthesis was higher in the controls than in the skin flap group, but at week 7 it was the reverse. The collagen synthesis was high and equal in both groups at weeks 2 and 3 and then decreased to weeks 4 and 5. Between weeks 6 and 7 a further decrease was found in the controls, but not in the skin flap group.

Animals↗
PubMed1990

Purification of an active gelatinase from collagen fiber fraction of granulation tissue in rats.

Gelatinase was extracted at 60 degrees C from the collagen fiber-rich fraction of granulation tissue induced by carrageenin in rats. A large part of the extracted gelatinase was unbound to Zn-chelating Sepharose. The unbound gelatinase gave a single band corresponding to a molecular mass of 57 kDa on SDS-substrate PAGE, but showed a much higher molecular mass (greater than 200 kDa) on Sephadex G-150 gel filtration. In addition, that unbound fraction contained gelatin fragments was revealed by SDS-PAGE. When the unbound fraction of Zn-chelating Sepharose was incubated at 37 degrees C, the gelatin fragments disappeared and the apparent molecular mass of gelatinase in gel filtration decreased. This gelatin degradation of the unbound fraction was enhanced by treatment with a 4-aminophenylmercuric acetate (APMA). The results suggest that the gelatinase is bound to gelatin fragments in the unbound fraction. After the treatment with APMA, the gelatinase was purified to to homogeneity; the purified gelatinase gave a single band corresponding to a molecular mass of 57 or 67 kDa on SDS-PAGE under nonreducing or reducing conditions, respectively. The purified gelatinase is a metalloproteinase, and extensively degraded gelatin, but showed no proteolytic activity toward alpha-casein or types I and IV collagens. The results suggest that the 67-kDa active gelatinase is bound to collagen fibers and plays an important role in a rapid degradation of collagen fibers in granulation tissue.

Animals↗
PubMed1979

Subcellular targets of the soluble SiO2-liberated macrophage factors in experimental granulation tissue.

The supernatant from SiO2-treated macrophages increased the incorporated radioactivity of collagen in the incubated experimental granulation tissue slices, especially in the rough endoplasmic reticulum fraction, markedly over that in the respective control preparation. The effect was observed after a 20 min incubation and increases linearly at least up to 3 h. The amount of RNA and phospholipids in the rough endoplasmic reticulum, calculated per protein, increased simultaneously in the incubated slices. The incorporation of the radioactive precursors into DNA in the slices and in the nuclei from proliferating granulation tissue was enhanced significantly by the soluble fraction from SiO2-treated macrophages. This effect could be seen after a 40 min incubation in the slices and after 5 min in the nuclei, when the incorporated radioactivity in DNA began to decrease in the control experiments.

Animals↗
PubMed1981

Effect of different doses of D-penicillamine and combined administration of D-penicillamine and methylprednisolone on collagen, glycosaminoglycans, DNA and RNA of granulation tissue, skin, bone and aorta in rats.

D-penicillamine (D-pen) in doses of 20, 100 and 500 mg/kg/day or D-pen 100 mg/kg/day plus methylprednisolone (MP) 2.0 mg/kg/day was administered daily for 42 days to rats implanted with viscose-cellulose sponges. Operated, pairfed rats served as controls. D-pen increased the DNA content of granulation tissue, but had no effect on the amount of tissue produced. In contrast, high dose D-pen reduced the content of DNA and collagen in skin. A dose related inhibition of collagen crosslink formation occurred in all tissues, particularly in skin, as indicated by increased proportions of extractable collagen with increased alpha/beta chain ratio and aldehyde content. Moreover, low doses of D-pen increased the hydroxyproline/proline ratio of acid soluble skin collagen, presumably due to solubilization of type III collagen as demonstrated by SDS-polyacrylamide gel electrophoresis in the presence of 3.6 M urea. These changes were associated with increased skin fragility and edema plus excess elastin deposition in the aorta after high dose D-pen treatment. Low dose D-pen stimulated the 35S-sulphate uptake into the sulphated glycosaminoglycans (GAGs) of granulation tissue without altering their relative amounts, whereas high dose D-pen reduced the concentration of chondroitin-4/6-sulphate in skin. MP antagonized the solubilizing effect of D-pen on collagen, probably by inhibition of the collagen synthesis. In addition, MP inhibited the cell proliferation and GAG metabolism. Food restriction reduced the DNA content of granulation tissue. The inhibitory effect of D-pen on the formation of granuloma collagen crosslinks in the presence of unaltered rate of collagen biosynthesis may diminish the amount of fibrotic tissue due to increased degradability of crosslink deficient collagen. Simultaneous administration of MP may facilitate this effect by inhibiting the biosynthesis of collagen. However, long-term D-pen treatment seems to increase the susceptibility of normal tissues to mechanical injury.

Animals↗
PubMed1991

Tracheal granulation tissue. A study of bacteriology.

We prospectively examined 19 patients (21 laryngotracheal reconstructions) over a 6-month period to evaluate the bacteriology of granulation tissue present at the time of Teflon stent removal and at the first laryngoscopy several weeks later. The most frequently recovered isolates were viridans streptococci, Pseudomonas aeruginosa, nonhemolytic Streptococcus, and Staphylococcus aureus. All but one positive culture were polymicrobial. The amount of tissue did not correlate with the duration of stenting and the amount of granulation tissue and number of organisms decreased after stent removal. Further prospective study of the most appropriate antimicrobial therapy is needed.

Bacteria↗
PubMedDate not supplied

[Immune complexes: mediators for the formation of inflammatory granulation tissue? Immunohistologic studies of the hyaline articular cartilage in chronic polyarthritis].

Previous reports describe the presence of immunoglobulins and complement components within rheumatoid articular cartilage, thereby suggesting an effect of immune complexes on the formation of pannus. This hypothesis is reinvestigated in this paper. As confirmed in our work, the superficial layer of rheumatoid hyaline cartilage may fulfill the immunohistological criteria for the presence of immune complexes. In osteoarthritis, however, a noninflammatory disease not mediated by immunologic mechanisms, similar results can be obtained. The presence of immune-proteins within hyaline cartilage therefore requires a cautious interpretation. Hyaline cartilage in rheumatoid arthritis is replaced by granulation tissue growing not only at its surface (pannus), but also in subchondral bone. We therefore also thoroughly investigated deep layers of hyaline cartilage in the vicinity of such subchondral tissue, but could not obtain any evidence for the presence of immune complexes therein. The growth of subchondral granulation tissue and the accumulation of PMN in the region of its junction with hyaline cartilage therefore appear to be independent of immune complexes within rheumatoid hyaline cartilage. It is suggested on the basis of these data that immune complexes possibly present in hyaline cartilage do not play an essential role in the formation of granulation tissue replacing cartilage in rheumatoid arthritis. It is, however, not entirely excluded that during advanced stages of rheumatic cartilage degradation immune complexes are formed within the matrix or carried into it from the extra-cartilaginous environment, and that they may then contribute to further cartilage destruction by enzyme release during phagocytic processes.

Antigen-Antibody Complex↗
PubMed2002

Use of a neodymium:yttrium-aluminum-garnet laser to remove exuberant granulation tissue from the esophagus of a horse.

Use of a neodymium:yttrium-aluminum-garnet laser to remove exuberant granulation tissue from the esophagus of a horse A 4-year-old Quarter Horse mare was evaluated because of recurrent esophageal obstruction. Endoscopic examination revealed a proliferative mass in the esophagus approximately 55 cm aborad to the nares. Histologic examination of biopsy specimens revealed diffuse suppurative esophagitis and granulation tissue with no evidence of neoplasia. The mass was excisedand ablated transendoscopically with a neodymium:yttrium-aluminum-garnet laser. During a follow-up examination 2 weeks after the final laser procedure, the mass was no longer apparent, and the esophagus appeared healed. Six months after the final examination, there had been no recurrence of the mass. In this horse, transendoscopic laser excision was efficacious and avoided problems associated with an esophagotomy.

Animals↗
PubMedDate not supplied

Wound matrix attachment regulates actin content and organization in cells of the granulation tissue.

Actin cytoskeletal polymerization is associated with a pro-proliferative, pro-survival state. We hypothesized that the actin polymerization of wound cells is increased in the presence of wound matrix attachment and is decreased after disruption of this attachment. Musculocutaneous flap and wound splinting models were used to investigate the effect of wound matrix attachment on the actin cytoskeleton. Disruption of wound matrix attachment was accomplished by incision of the wound matrix/dermis interface (wound matrix release) and/or desplinting. Polymerized actin was assayed with phalloidin labeling of wound specimens 24 hours after disruption of attachment and a method to quantify the content and organization of polymerized actin in granulation tissue was used. Disruption of wound matrix attachment decreased the content of polymerized actin, the actin staining intensity, and the actin fiber organization in the granulation tissue of both the flap and splint models. Disruption of wound matrix attachment decreased actin polymerization and fiber organization in the granulation tissue. Our data support the concept that the state of wound matrix attachment regulates the actin cytoskeleton of wound cells.

Actins↗
PubMed1987

Epidermal growth factor increases granulation tissue formation dose dependently.

Recent studies have shown that epidermal growth factor (EGF) stimulated the rate of formation of granulation tissue in a model of wound repair (A. Buckley, et al., Proc. Nat. Acad. Sci. USA 82: 7340, 1985). Because pharmacologic doses of EGF were used previously, the relationship of EGF concentration to physiologic effects was determined in this study. Rats were implanted with subcutaneous polyvinyl alcohol sponges containing slow-release pellets formulated to release 0, 0.1, 1.0, or 10 micrograms of EGF/day. Tissue response was judged by the degree of histologic organization and vascularity, as well as several quantitative parameters: wet weight, hydroxyproline content, protein content, and DNA concentration. Each of these parameters showed consistent increases by Day 5 after implantation, when inflammation and edema had subsided. Compared with placebo controls, hydroxyproline (collagen) content was significantly increased by as little as 1 microgram/day of EGF, and DNA content was significantly increased by all dose levels of EGF. Endogenous EGF concentration in experimental granulation tissue was found to be fairly constant (30-40 ng/g wet wt); however, the increasing cellularity of the sponges may have reduced the local concentration of free EGF to low levels. Pellets releasing as little as 4 ng/hr of EGF into the surrounding tissue were able to accelerate wound healing, suggesting that the availability of this growth factor may be a rate-limiting step in wound repair.

Animals↗
PubMed2003

Abscess-forming inflammatory granulation tissue with Gram-positive cocci and prominent eosinophil infiltration in cats: possible infection of methicillin-resistant Staphylococcus.

We occasionally encounter feline cervical or mesenteric lesions diagnosed histopathologically as abscess or inflammatory granulation tissue with eosinophil infiltration. Gram-positive cocci accompany the lesions. In the present study, such lesions obtained from 27 cats were examined to evaluate the histopathologic features and the nature of the causative bacteria. The average age was 7.3 +/- 3.5 years. No sex predilection was observed. Most frequent locations of the lesions included the abdominal cavity with/without mesenteric lymph nodes (11/27, 41%) and subcutaneous tissue or lymph nodes of the neck (9/27, 33%). Common clinical presentation was a localized mass. Grossly, the lesions contained abscesses in the center and were surrounded by fibrous tissue. Microscopically, the necrotic zone contained bacterial colonies. Large numbers of eosinophils and macrophages infiltrated the area surrounding the necrotic tissue. The surrounding connective fiber-rich granulation tissue demarcated the eosinophilic abscess. The bacteria were Gram-positive cocci in 23 of the 27 cats and were positive for anti-staphylococcus antiserum in 19 of the 23 cats. In 15 out of 17 lesions, the colonies expressed immunoreactivity to penicillin-binding protein 2', which is a drug-resistance gene product of methicillin-resistant Staphylococcus (MRS) species. These findings suggest strongly that MRS causes this type of infectious lesion.

Abdominal Abscess↗
PubMed2001

Overexpression of hepatocyte growth factor/scatter factor promotes vascularization and granulation tissue formation in vivo.

The effect of hepatocyte growth factor/scatter factor (HGF/SF) during wound healing in the skin was investigated, using HGF/SF-overexpressing transgenic mouse model. Histological analysis of HGF/SF transgenic mouse excisional wound sites revealed increased granulation tissue with marked vascularization. Northern blot analysis demonstrated that, relative to control, vascular endothelial growth factor (VEGF) expression in transgenic skin was significantly higher at baseline and was robustly up-regulated during wound healing. Elevated levels of VEGF protein were detected immunohistochemically, predominantly in endothelial cells and fibroblasts within the granulation tissue of HGF/SF transgenic skin. Serum levels of VEGF were also elevated in HGF/SF transgenic mice. Thus, results from our study suggest that HGF/SF has a significant effect on vascularization and granulation tissue formation during wound healing in vivo, involving with induction of VEGF.

Animals↗
PubMed2001

Effects of varying levels of subatmospheric pressure on the rate of granulation tissue formation in experimental wounds in swine.

The use of subatmospheric pressure to promote wound healing has increased in popularity during the last several years. The original studies on granulation tissue formation used a 125-mmHg vacuum. The use of alternative sources of subatmospheric pressure has led to many questions regarding efficacy or risk. In this report a swine model is used to quantify and compare the effects of low vacuum suction (25 mmHg) and high vacuum suction (500 mmHg) produced by various vacuum pumps and wall suction systems with the standard 125-mmHg vacuum. Additionally, the effects of an unregulated air leak in the sealing system were examined. All four wound treatments were examined on each of 4 pigs. Wounds were treated until one of the wounds had granulated to a level flush with the surrounding tissue. Wounds treated with the standard 125-mmHg vacuum had filled with granulation tissue by day 8. At this time wounds treated with 25 mmHg had filled 21.2% with new granulation tissue, and wounds treated with 500 mmHg had filled 5.9% with new tissue. Wounds treated with 125 mmHg with a hole in the sealing drape had increased in size 197% because of the debridement of necrotic tissue. In conclusion, wounds treated with a 125-mmHg vacuum exhibited a significant (p < 0.0001) increase in the rate of granulation tissue formation compared with treatment at 25 mmHg or 500 mmHg. The presence of an unregulated air leak in the sealing drape results in significant progression (p < 0.0001) of the wound secondary to dehydration and progressive necrosis.

Animals↗
PubMed1999

Cloning of cDNA for rat pro alpha1(V) collagen mRNA. Expression patterns of type I, type III and type V collagen genes in experimental granulation tissue.

A cDNA clone for rat pro alpha1(V) collagen mRNA was constructed using PCR amplification, with primers based on human and hamster COL5A1 gene sequences. The clone pRCVA1 is 560 nucleotides long and it encodes for the carboxy propeptide of type V procollagen. Homology shared with type I collagen sequence was 64%, with type II collagen 65% and with type III collagen 61%. To evaluate the spatial and temporal expression of type V collagen mRNA in wound healing model, subcutaneously implanted viscose cellulose sponges in rats were used to induce granulation tissue formation. Analyses on granulation tissue were carried out on days 5, 8, 14, 21, 30, 59 and 84. Specific cDNA probes to pro alpha1(I), pro alpha1(III) and pro alpha1(V) collagen mRNA were used in slot blot, Northern and in situ hybridization. Type I collagen gene expression was upregulated at the initial stage of wound healing, type III collagen gene expression was constant and from the day 14 onwards type I and III collagen gene expressions were at the same level. Type V collagen gene expression was seen at every time point studied but at a considerably lower level than type I and III collagens. In situ hybridization showed that type V collagen was expressed in two different cell types. In conclusion, type V collagen was expressed in the wound healing model from at least day 5 onwards and it was synthesized by fibroblast-like and rounded cells.

Actins↗
PubMed2000

Expression and activity of matrix metalloproteinase-2 and -9 in experimental granulation tissue.

The restoration of functional connective tissue is a major goal of the wound healing process which is probably affected by matrix-modifying enzymes. To evaluate the spatial and temporal expression of matrix metalloproteinases (MMP) MMP-2 and MMP-9 and to study the regulation of MMP-2 in wound healing, subcutaneously implanted viscose cellulose sponges in rats were used to induce granulation tissue formation for up to 3 months. MMP-2 mRNA expression was seen throughout the experiment and it was highest after 2 months. MMP-9 gene expression was low between days 8-21 and increased after 4 weeks of granulation tissue formation. Membrane-type 1 MMP (MT1-MMP) mRNA was upregulated early and tissue inhibitor 2 of MMP (TIMP-2) mRNA later during wound healing. In in situ hybridization the expression of MMP-2 mRNA was seen mostly in fibroblast-like cells and MMP-9 mRNA in macrophage-like cells. MMP-9 immunoreactivity was detected in the polymorphonuclear leukocytes and macrophage-like cells on days 3-8. MMP-9 proteolytic activity was observed only on days 3-8. The active form of the MMP-2 increased up to day 14, whereafter it remained at a constant level, whereas latent MMP-2 did not show any apparent changes during the experimental period. We conclude that MMP-2 is important during the prolonged remodelling phase, whereas the gelatinolytic activity of MMP-9 was demonstrated only in early wound healing, and the MMP-9 gene is upregulated when the granulation tissue matures.

Animals↗
PubMed2003

Neutralization of hepatocyte growth factor leads to retarded cutaneous wound healing associated with decreased neovascularization and granulation tissue formation.

To elucidate biologic functions of hepatocyte growth factor and the c-Met receptor in cutaneous wound healing, we analyzed expression and localization of hepatocyte growth factor and c-Met receptor and used a strategy to neutralize endogenous hepatocyte growth factor in a cutaneous wound healing model in mice. Following excision of full-thickness skin on the dorsum of mice, expression of both hepatocyte growth factor and the c-Met receptor increased transiently in cutaneous tissues. Expressions of hepatocyte growth factor increased as early as 2 d postwounding and reached a peak on day 2, whereas the c-Met receptor expression reached a peak 2-4 d postwounding. Immunolocalization of the c-Met receptor indicated that c-Met receptor expression was upregulated in keratinocytes, vascular endothelial cells, and myofibroblasts in granulation tissue, hence these are potential target cells of hepatocyte growth factor. When normal rabbit IgG or neutralizing anti-hepatocyte growth factor IgG was locally and continuously delivered to subcutaneous lesions, the number of capillary vessels decreased with the neutralization of hepatocyte growth factor and there was an associated decreased expansion of granulation tissue. Likewise, retardation in re-epithelialization and the rate of wound closure occurred with neutralization of endogenous hepatocyte growth factor on days 4 and 7 postwounding. Therefore, hepatocyte growth factor is definitely involved in enhancing cutaneous wound healing processes, including re-epithelialization, neovascularization, and granulation tissue formation.

Animals↗
PubMed1993

Changes in cell proliferative activity in granulation tissue invading the root canal.

Root canal models were implanted in rats in order to investigate histologically the movement of granulation tissue invading apical canals and dead spaces and changes in cell proliferative activity as indicated by 3H-thymidine. The models were prepared to have 1, 2, and 3 mm apical canals, with and without dead spaces, to simulate preparations short of the apex and obturation to several levels. After 12 wks implantation of the models without dead spaces, granulation tissue invading 1 mm apical canals did not degenerate and cell proliferative activity remained high. However, tissue invading the 2 and 3 mm apical canals tended to be necrotic and cell proliferative activity was decreased. In the models with dead spaces, the tissue in the 1 and 2 mm apical canals developed and invaded the dead spaces, and still possessed proliferative activity 12 wks after implantation. In contrast, the tissue in the 3 mm apical canals did not invade the dead spaces, even after 12 wks, and no proliferative activity was observed.

Animals↗