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The experimental basis of intestinal suturing. Effect of surgical technique, inflammation, and infection on enteric wound healing.

Factors that incite inflammation at the healing wound prolong the lag period of wound healing and delay the return of strength at the suture line. Inflammation activates bowel-wall collagenase, which degrades the collagen within the wound, eroding the foundation in which sutures are anchored. Experimental studies have compared the impact of various surgical techniques. Sutures placed by hand uniformly invoke an inflammatory response because dragging the thread through the bowel wall injures tissue. Single-layer anastomoses heal more rapidly than double-layer suture lines. The inner layer causes avascular necrosis of the inverted cuff. Experimental studies have not clearly shown the superiority of inverting suture lines over everting ones. Experimental studies done over the last century indicate that the single-layer inverting anastomosis recommended by Lembert and Halstead adequately compensates for enteric wound weakness during the lag period. Other techniques of sewing an anastomosis provide no clear advantage. Other factors that incite inflammation also delay enteric wound healing. Debris, necrotic tissue, or infection illicit an inflammatory response with detrimental effects on the anastomosis. Antibiotics, by assisting in the control of infection or by minimizing the size of an inoculum, help speed healing. Stapling devices violate many of the doctrines of intestinal suturing. Experimental studies suggest, however, that staple lines incite a minimal inflammatory response. Consequently, wounds closed with stapling devices regain strength more rapidly than those closed with traditional surgical techniques.

Animals↗

Synthesis of type I collagen in healing wounds in humans.

To quantify wound healing in surgical patients, samples of wound fluid were collected through a silicone rubber tube for 7 postoperative days and their concentrations of the carboxyterminal propeptide of type I procollagen (PICP) and the aminoterminal propeptide of type III procollagen (PIIINP) were measured with specific radioimmunoassays. The mean concentration of PICP in would fluid on day 1 was 207 +/- 92 (SD) micrograms/L, and on day 2 908 +/- 469 micrograms/L (p less than 0.001, signed rank test). On day 7, the mean concentration reached was 380 times higher than that of day 1 (79,330 +/- 54,151 micrograms/L). Only one peak of PICP antigenicity, corresponding to the intact propeptide as set free during synthesis of type I procollagen, was detected on Sephacryl S-300 gel filtration analysis of wound fluid samples. The mean concentration of PIIINP was 70 +/- 61 micrograms/L on day 1, 86 +/- 88 micrograms/L on day 2, and 180 +/- 129 micrograms/L on day 3 (p less than 0.001 when compared with day 1). Finally on day 7, a 250-fold concentration (17,812 +/- 9839 micrograms/L), compared with day 1, was reached. Methods described in the present paper allow separate and repetitive quantification of the synthesis of both type I and type III procollagen during human wound healing.

Adult↗

Overview of matrix metalloprotease modulation and growth factor protection in wound healing. Part 1.

Increasingly, basic research is revealing that the biochemical environment of the non-healing wound is different from that of the healing wound in ways that negatively affect multiple aspects of the healing process. These areas of biochemical difference may provide avenues of research for new therapeutic interventions. A particular biochemical finding in nonhealing wounds of diverse etiologies is the persistent elevated levels of a specific class of proteolytic enzymes known as the matrix metalloproteases or MMPs. The effects of these high levels of MMPs in the wound environment are thought to include local destruction of growth factors and their receptors as well as degradation of granulation tissue components. MMPs are fast gaining attention as potential causative factors in impaired healing, and recently, strategies have been focused on preventing or reducing their levels, once elevated, in the wound environment. A proprietary combination of bovine collagen and oxidized regenerated cellulose has demonstrated an ability to not only reduce elevated wound proteases but also sequester and protect wound growth factors and promote cellular chemotaxis and proliferation.

Cell Communication↗

Wound healing and chronic wounds.

A renaissance is taking place in the field of wound healing. Interest in wounds has probably never been so great, and a number of key therapeutic advances in the last 2 or 3 years are only an indication of more progress to come. In large part, we are benefitting from decades of basic research that has provided the knowledge and technical advances for many of today's therapeutic products. In this manuscript, we will provide the initial framework for wound healing and have summarized some key concepts.

Biocompatible Materials↗

A new concept of a multidisciplinary wound healing center and a national expert function of wound healing.

HYPOTHESIS: An independent, multidisciplinary wound healing center in an accepted national expert function of wound healing is the optimal way to improve prophylaxis and treatment of patients with problem wounds. DESIGN: A clinical perspective analysis. SETTING: An independent, multidisciplinary wound healing center focusing on all types of problem wounds, organized as a university hospital department, and integrated in an expert function in the national health care organization of Denmark. PATIENTS AND METHODS: Patients with all types of problem wounds referred to and treated in the center during the first years of its existence provided a model for a new multidisciplinary structure for treatment of wound patients in the health care system. RESULTS: During the first 3 years of the fully functioning wound healing center, a total of 23 802 patient consultations were performed in the outpatient clinic, and 1014 patients with problem wounds were hospitalized in the inpatient ward. The surgical concept of the center has resulted in improved healing rates in patients with leg ulcers and decreased rates of major amputations. The outpatient function has resulted in a decrease in the number of patients transported in beds to the center. This structure provides better opportunities for basic and clinical research as well as for establishing expert education for all types of health care personnel. The center's structure has been the background for establishing an expert function in wound healing, allowing the wound healing area area to be fully integrated in the Danish National Health Care System. Overall, the concept and structure of the center have enhanced the knowledge and understanding of wound problems and increased the status of wound healing and patient care. CONCLUSIONS: Establishing multidisciplinary centers integrated into an accepted national expert function of wound healing is an optimal way to improve the clinical outcome of prophylaxis and treatment of all types of problem wounds. This model, with minor adjustments, may be applicable for both industrialized and developing countries.

Algorithms↗

Co-expression of p16INK4A and laminin 5 by keratinocytes: a wound-healing response coupling hypermotility with growth arrest that goes awry during epithelial neoplastic progression.

The replicative lifespan of human keratinocytes in culture is restricted by a telomere-unrelated induction of p16INK4A (p16) and p14ARF. We have found that, in vivo, p16 is expressed by epidermal and oral keratinocytes at the migrating fronts of healing wounds and at the stromal interface of severely dysplastic and early invasive lesions and that such cells also invariably display increased expression of Laminin 5 (Lam5). In culture, p16 and Lam5 are coexpressed in keratinocytes at senescence, at the edges of wounds made in confluent cultures, and when cells are plated on dishes coated with the gamma2 precursor form of Lam5 (Lam5gamma2pre). Lam5/p16 coexpression in all three in vitro settings is associated with directional hypermotility and growth arrest. Hypermotility and growth arrest are uncoupled in p16- and p14ARF/p53-deficient keratinocytes and squamous cell carcinoma (SCC) cells; such cells become hypermotile is response to Lam5gamma2pre but do not growth arrest. Thus, the Lam5/p16 response is activated in normal wound healing, causing growth arrest of migratory keratinocytes that lead wound reepithelialization. This response also becomes activated at a critical stage of neoplastic progression, acting as a tumor suppressor mechanism. Rare premalignant cells that lose p16 remain motile and proliferative, thereby resulting in invasive growth as SCC.

Carcinoma, Squamous Cell↗

Common complications of wound healing. Prevention and management.

The culmination of any operation is a healed wound. Failure of a wound to heal increases time spent in the hospital and the expense, and may start a cascade of progressive complications. Thus, it is important to optimize as many factors as possible before operation and to have the knowledge and resources to deal with wound problems should they occur. This approach involves understanding basic wound healing with its multiple factors, including nutrition, and knowing how to deal with potential adverse factors in wound healing, such as chemotherapy, corticosteroids, and radiation.

Bandages↗

Fetal cleft lip repair in lambs: histologic characteristics of the healing wound.

Sequential surgical procedures in cleft lip/palate (CL/P) patients result in scar formation that is believed to be associated with midface growth retardation. By use of a previously developed fetal lamb model, wound healing characteristics were investigated after in utero CL repair. It is hypothesized that scarless healing after fetal CL repair occurs without inflammation and scar formation. CL wounds were created in mid-second-trimester fetal lambs and either repaired in three layers (mucosa, muscle, and skin) or left unrepaired. Fetuses were then harvested at 7, 14, and 21 days postoperatively, and the wound site was examined microscopically. When created at 75 days' gestation (term = 145 days), fetal lamb CL wounds heal rapidly without inflammation and scar formation. By day 21 postoperatively, there was complete regeneration of skin, muscle, and mucosa, as well as epidermal appendages. With this model, it will be possible to compare the effects of scarless fetal CL repair with those of postnatal repair on midface growth.

Animals↗

Effect of macrophage stimulation on collagen biosynthesis in the healing wound.

Immunomodulators that enhance macrophage function have been shown to be beneficial in a number of wound-healing models in humans and in experimental animals. The exact mechanism of this improved healing is unclear. To assess the role of collagen biosynthesis, the immunomodulator glucan phosphate was utilized in two murine models of wound healing, i.e., colon anastomosis and full-thickness skin incision. Tensile strength was evaluated using computer-assisted constant velocity tensiometry. Collagen biosynthesis was determined by assaying hydroxyproline content of wound hydrolysates by N-(9-fluorenyl)methoxycarbonyl/o-phthalaldehyde high-performance liquid chromatography. Experimental animals were treated with (1-3)-beta-D-glucan phosphate (250 mg/kg) intravenously 24 hours prior to colon anastomosis or skin incision. A second dose of glucan phosphate was given immediately postoperatively. Control animals received dextrose and water (5% w/v) intravenously. Tensile strength and hydroxyproline content were measured on postoperative Day 3. In the skin wound model, glucan phosphate treatment increased (P < 0.05) tensile strength by 42 per cent (342.5 +/- 12.2 vs 241.8 +/- 4.8 g), and hydroxyproline content was increased by 23.5 per cent (242.0 +/- 14.4 vs 196.8 +/- 10.5 pmol/microg; P < 0.05). In the glucan phosphate group, colon tensile strength was significantly (P < 0.05) increased by 34 per cent (34.2 +/- 2.3 g vs 45.8 +/- 2.1 g), and hydroxyproline content was increased by 7 per cent (47.45 +/- 3.31 vs 44.34 +/- 3.74 pmol/microg). These data indicate that macrophage modulation with glucan phosphate will increase tensile strength in experimental colon and skin wounds. In addition, we observed a positive correlation between glucan phosphate treatment, wound tensile strength, and collagen biosynthesis.

Adjuvants, Immunologic↗

The influence of cortisol on wound healing of the skin and distant connective tissue response.

The effect of moderate dosage cortisol treatment on healing wounds and on the response in skin distant to the wounds was investigated. The stress-strain characteristics of a skin wound and intact skin specimens were examined. Also, skin thickness, water content and collagen content were measured. Cortisol treatment resulted in reduced extensibility and increased stiffness for ten day wounds, but unchanged failure energies; for 20 day wounds, failure energies were reduced, indicating reduced ability to withstand rupturing forces. Thus, the mechanical properties of healing wounds under moderate dosage, long term corticosteroid treatment were found to be slightly impaired. A systemic effect of wounding at day 10 resulted in decreased stiffness and increased collagen content of the skin distant to the wound.

Animals↗

The effect of indirect injury to peripheral nerves on wound healing after firearm wounds.

This paper reports the pathologic and functional changes of the sciatic nerve indirectly injured by firearms and its effect on wound healing. Twelve dogs were used. Their hind legs were shot with 1.03-gram steel spheres, avoiding direct injury to the sciatic nerve. The experimental animals were divided into two groups according to the impact velocity, that is, a 1,500 m/sec high velocity projectile (HVP) group and a 400 m/sec, low velocity projectile (LVP) group as a control. After wounding, the same procedure was followed in all dogs. At intervals, the tissues of the wound track were excised to determine the level of cyclic nucleotides (cAMP, cGMP), secondary messengers of neurotransmitters, the activity of acetylcholinesterase (AChE), and the content of hydroxyproline, which indicates the degree of wound healing. The sciatic nerve next to the wound track was excised for light and electron microscopic study. The neurofilament in the sciatic nerve was examined by immunohistochemistry with monoclonal neurofilament-200 antibody. The results showed that the indirect injuries to the sciatic nerves of the HVP group were more severe than in those in the LVP group and included naked axons, stripped myelin sheaths, widened nodes of Ranvier, and decrease in the numbers of neurofilaments. At 6 hours, cAMP content in the HVP group was significantly lower than that in the LVP group. It began to rise and approached to the level of LVP group at 10 days. The cGMP content, activity of AChE, and hydroxyproline content of the HVP group were significantly lower than those of the LVP group. The results suggested that the indirect injury to sciatic nerves, especially to the cytoskeleton, after HVP wounds may induce the disturbance of an axoplasmic flow and influence the metabolism of trophic factors such as cAMP, cGMP, and AChE. All changes could contribute to the delay in wound healing.

Acetylcholinesterase↗

Macrophage-derived growth factors in wound healing: regulation of growth factor production by the oxygen microenvironment.

There is a large amount of current research investigating the pathophysiologic aspects of pulmonary hypertension. Results of this research indicate the potential role of the macrophage as one possible mediator of pulmonary hypertension. The macrophage has been shown to produce numerous soluble mediators, including various growth factors, some of which may be involved in the cellular proliferation of the arterial wall. Most of the knowledge regarding oxygen microenvironments, macrophages, and growth factor production comes from research on the healing wound. A considerable amount of what has been learned about the role of the macrophage in cutaneous wound healing may have direct applicability to macrophage function in acute lung injury and repair, and pulmonary hypertension. Macrophages play a pivotal role in cutaneous wound repair. They participate in immunologic and nonspecific host defense responses and produce numerous growth factors that regulate mesenchymal cell proliferation, migration, and synthesis of extracellular matrix proteins. These same responses are probably at work during the progression of pulmonary hypertension.

Growth Substances↗

Human wound healing fibroblasts have greater contractile properties than dermal fibroblasts.

Contractile and phenotypic properties of human fibroblasts from healing wounds were compared to those of dermal fibroblasts using in vitro models. Wound fibroblasts were recovered from implants, made of a polyvinyl alcohol sponge threaded into a perforated silicone tube, 12 days after their subcutaneous implantation in human volunteers. Dermal fibroblasts were isolated from the skin of healthy subjects. Two morphologically different fibroblast populations were observed in cells cultured from implants. In order to characterize these fibroblast populations, intracellular alpha-actin expression was studied by immunofluorescence labeling of cells cultured in monolayer. This protein was detected in less than 1% of the dermal fibroblasts. By contrast, 30 to 40% of wound fibroblasts were labeled and contained fiber networks of alpha-actin. These results confirm the presence of myofibroblasts in human wound healing tissues. The contractile property of fibroblasts and myofibroblasts was evaluated using a three-dimensional cell culture model (fibroblast populated collagen gels). Cells were incorporated in a collagen matrix and cultured for 14 days. The surface area of collagen gels was measured every day. Our results show that wound fibroblasts strongly contract collagen gels during the first 24 hr (surface area at 24 hr = 20-55% of initial surface area) in comparison to dermal fibroblasts (surface area at 24 hr = 70-75% of initial surface area). This superior level of contraction was observed until the fifth day of culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗

Selective loss of blood group antigens during wound healing.

Frozen sections of healing wounds of the oral mucosa of 3 rhesus monkeys were examined by a double layer immunofluorescent technic for the presence of blood B-like antigens. Antigen activity was present in normal epithelium and in the epithelial outgrowth into the wounds but was absent from the outgrowth following treatment with lipid solvents. This finding suggests that glycoprotein but not glycolipid associated antigen is lost from the surface of epithelial cells during wound healing, a pattern of selective loss which may reflect a difference between regenerative and neoplastic lesions.

Animals↗

The effects of a broad-spectrum matrix metalloproteinase inhibitor on characteristics of wound healing.

This study investigates the effects of a broad-spectrum matrix metalloproteinase inhibitor (MMP-i) on the rate of closure, hydroxyproline deposition, and macrophage infiltration in healing wounds. Full-thickness excisional wounds were created on the dorsal surface of hairless mice. Two experimental groups were used to measure rates of wound closure: (a) MMP-i administration (0.03, 0.3, 3.0, and 30 microg/mL) on days 0-1 postwounding (inflammatory phase) and (b) MMP-i administration (0.03, 0.3, 3.0, and 30 microg/mL) on days 6-8 postwounding (proliferative phase). Additionally, hydroxyproline deposition and percent macrophage infiltration were measured in skin wound margins on days 2, 8, and 16 postwounding. MMP-i administration at concentrations of 0.03, 0.3, and 3.0 microg/mL on days 0-1 postwounding significantly (p <.05) increased the rate of wound closure. No significant effect on the rate of wound closure was observed with MMP-i administration on days 6-8 postwounding. Hydroxyproline deposition was significantly (p <.05) increased on day 8 postwounding, and the percent macrophage infiltration was significantly (p <.05) decreased on day 2 postwounding by MMP-i administration on days 0-1 postwounding. These experiments demonstrate that MMP-i administration during the inflammatory phase significantly affects several characteristics of wound healing. We postulate that these effects may be attributed to decreased degradation of ECM components, increased concentrations of endogenous growth factors, and a shortened inflammatory phase.

Animals↗