The Effect of Comorbidities on Wound Healing.

Wound healing is affected by several factors. Preexisting diagnoses may significantly alter, delay, or inhibit normal wound healing. This is most commonly seen with chronic disorders, such as diabetes and renal failure, but also occurs secondary to aging and substance abuse. Less commonly, genetic or inflammatory disorders are the cause of delayed wound healing. In some cases, it is not the illness, but the treatment that can inhibit wound healing. This is seen in patients getting chemotherapy, radiation, steroids, methotrexate, and a host of other medications. Understanding these processes may help treat or avoid wound healing problems.

Arginine-Dual roles as an onconutrient and immunonutrient.

Arginine is an important player in numerous biologic processes and studies have demonstrated its importance for cellular growth that becomes limiting in states of rapid turnover (e.g., malignancy). Thus, arginine deprivation therapy is being examined as an adjuvant cancer therapy, however, arginine is also necessary for immune destruction of malignant cells. Herein we review the data supporting arginine deprivation or supplementation in cancer treatment and the currently registered trials aimed at understanding these divergent strategies. J. Surg. Oncol. 2017;115:273-280. © 2016 Wiley Periodicals, Inc.

SDF-1α mediates wound-promoted tumor growth in a syngeneic orthotopic mouse model of breast cancer.

Increased growth of residual tumors in the proximity of acute surgical wounds has been reported; however, the mechanisms of wound-promoted tumor growth remain unknown. Here, we used a syngeneic, orthotopic mouse model of breast cancer to study mechanisms of wound-promoted tumor growth. Our results demonstrate that exposure of metastatic mouse breast cancer cells (4T1) to SDF-1α, which is increased in wound fluid, results in increased tumor growth. Both, wounding and exposure of 4T1 cells to SDF-1α not only increased tumor growth, but also tumor cell proliferation rate and stromal collagen deposition. Conversely, systemic inhibition of SDF-1α signaling with the small molecule AMD 3100 abolished the effect of wounding, and decreased cell proliferation, collagen deposition, and neoangiogenesis to the levels observed in control animals. Furthermore, using different mouse strains we could demonstrate that the effect of wounding on tumor growth and SDF-1α levels is host dependent and varies between mouse strains. Our results show that wound-promoted tumor growth is mediated by elevated SDF-1α levels and indicate that the effect of acute wounds on tumor growth depends on the predetermined wound response of the host background and its predetermined wound response.

Excessive nitric oxide impairs wound collagen accumulation.

BACKGROUND: Nitric oxide (NO) plays a major regulatory role in wound collagen synthesis. We hypothesized that this regulatory role is tightly controlled by the levels of NO in the wound environment and that supranormal wound NO generation impairs wound collagen accumulation. MATERIALS AND METHODS: We used the model of turpentine-induced granuloma in male Sprague-Dawley rats as a sterile inflammatory stimulus generating large amounts of NO. In this environment, NO generation increased by 260%, whereas collagen deposition was significantly reduced by 38.5% (729.7 ± 81.5 versus 449.4 ± 76.3 µg hydroxyproline/100 mg sponge, P<0.05). Inhibition of NO synthase activity using 300 mM L-N6-(1-iminoethyl)-lysine, a highly potent and selective inhibitor of inducible NO synthase, significantly reduced NO elevation by 43.3% and increased wound collagen deposition by 37.3% (P<0.05). These effects occurred without any anti-inflammatory effects of L-N6-(1-iminoethyl)-lysine as assessed by the white blood cell counts and levels of interleukins 1 and 6. CONCLUSIONS: The data show that high levels of NO within the wound environment significantly reduce wound collagen deposition. Inhibition of NO generation restores collagen levels to normal levels. The regulatory effects of NO on wound collagen appear to be highly correlated with the amount of NO generated.

Arginase inhibition promotes wound healing in mice.

OBJECTIVE: Arginase plays important regulatory roles in polyamine, ornithine, and nitric oxide syntheses. However, its role in the healing process has not been delineated. In this study, we used a highly potent and specific inhibitor of arginase, namely 2(S)-amino-6-boronohexanoic acid NH4 (ABH) to evaluate the role of arginase function in wound healing. MATERIALS AND METHODS: ABH or saline was applied topically to full thickness, dorsal, excisional wounds in C57BL/6 mice every 8 hours for 14 days post surgery and the rate of wound closure was estimated planimetrically. Wound tissue was harvested from mice sacrificed on postoperative days 3 and 7 and examined histologically. The extent of epithelial, connective, and granulation tissue present within the wound area was estimated histomorphometrically. The effect of ABH on wound arginase activity, production of nitric oxide metabolites (NO(x)), and presence of smooth muscle actin positive cells (myofibroblasts) was evaluated. RESULTS: While arginase activity was inhibited in vivo, the rate of wound closure significantly increased 7 days post-surgery, (21 ± 4%: P < .01; Student t test) in ABH treated animals. This was accompanied by an early increase in wound granulation tissue and accumulation of NO(x) followed by enhanced re-epithelialization and localization of myofibroblasts beneath the wound epithelium. CONCLUSION: Arginase inhibition improves excisional wound healing and may be used to develop therapeutics for early wound closure.

Peptide inhibitors of MK2 show promise for inhibition of abdominal adhesions.

BACKGROUND: Abdominal adhesions are a common side effect of surgical procedures with complications including infertility, chronic pain, and bowel obstruction, which may lead to the need for surgical lyses of the adhesions. Mitogen-activated protein kinase-activated protein kinase 2 (MK2) has been implicated in several diseases, involving inflammation and fibrosis. Thus, the development of a cell-penetrating peptide (CPP) that modulates MK2 activity may confer therapeutic benefit after abdominal surgery in general and more specifically after bowel anastomosis. METHODS: This study evaluated the function of a CPP inhibitor of MK2 in human mesothelial cells and in a rat bowel anastomosis model. To determine IC50 and basic specificity, kinase inhibition was performed using a radiometric assay. Enzyme-linked immunoassay (ELISA) was used to evaluate interleukin-6 (IL-6) expression in response to IL-1ß and tumor necrosis factor-α (TNF-α) stimulation in vitro to validate MK2 kinase inhibition. Following bowel anastomosis (10 rats for each control and treatment at 4 and 10 d), the rats were evaluated for weight loss, normal healing (colonic burst strength and hydroxyproline content at the anastomosis), and number and density of adhesions. RESULTS: The IC50 of the MK2 inhibitor peptide (22 µM) was similar to that of the nonspecific small molecule rottlerin (IC50 = 5 µM). The MK2 inhibitor peptide was effective at suppressing IL-1ß and TNF-α stimulated IL-6 expression in mesothelial cells. In vivo, the MK2 inhibitor peptide was effective at suppressing both the density and number of adhesions formed as a result of bowel an anastamosis. Importantly, the peptide had no negative effect on normal healing. CONCLUSIONS: In conclusion, the peptide inhibitor of MK2, MMI-0100, has the potential to significantly reduce inflammation through suppression of inflammatory cytokine expression and showed promise as a therapeutic for abdominal adhesions.

Nutrition and wound healing: an update.

Nutrition has always been noted to be one of the major influences on the successful outcome of wound healing. The exuberant cellular and biochemical events that constitute the wound-healing cascade require energy, amino acids, oxygen, metals, trace minerals, and vitamins for successful completion. Many nutritional deficiencies impact on wound healing by impeding fibroblast proliferation, collagen synthesis, and epithelialization. There are also nutrients that can enhance wound-healing responses. It is imperative for physicians to obtain a complete nutritional history and consider nutritional intervention as a means of affecting the course of healing. This review examines many of the advances that have occurred in understanding nutrition/wound interactions.

Intra-abdominal healing: gastrointestinal tract and adhesions.

The abdominal cavity represents one of the most active areas of surgical activity. Surgical procedures involving the gastrointestinal (GI) tract are among the most common procedures performed today. Healing of the GI tract after removal of a segment of bowel and healing of the peritoneal surfaces with subsequent adhesion formation remain vexing clinical problems. Interventions to modify both the responses are myriad, yet a full understanding of the pathophysiology of these responses remains elusive. Different aspects of GI and peritoneal healing, with associated factors, are discussed in this article.

Laparoscopic surgery improves blood glucose homeostasis and insulin resistance following distal gastrectomy for cancer.

BACKGROUND: Prevention of blood glucose elevation and insulin resistance could be more pronounced in patients undergoing laparoscopic rather than open gastrectomy. METHODS: Fifty-seven patients underwent distal gastrectomy by either laparoscopy (n = 36) or an open approach (n = 21). Blood glucose, serum insulin, and the daily insulin secretion rate (urinary C-peptide) were measured. Insulin resistance was evaluated using an adapted homeostasis model assessment of insulin resistance (HOMA-R). RESULTS: Blood glucose levels were lower in the laparoscopy group than in the open group on the operative day and on postoperative days (POD) 1 and 3 (P < .001, P = .001, and P = .024, respectively). Serum insulin levels were lower in the laparoscopy group than in the open group on POD 1 and 3 (P = .045 and P = .027, respectively). HOMA-R was lower in the laparoscopy group than in the open group on POD 1 and 3 (P = .024 and P = .009, respectively). Daily insulin secretion rates were lower in the laparoscopy group than in the open group on POD 1 (P = .023). CONCLUSIONS: Laparoscopic surgery prevents blood glucose elevation and improves insulin resistance compared with open surgery.

Acute wounds accelerate tumorigenesis by a T cell-dependent mechanism.

We investigated the influence of acute wounding on tumor growth in a syngeneic mouse breast cancer model. Metastatic mouse breast cancer cells (4T1) were orthotopically injected into the mammary fat pads of BALB/c mice, and animals were wounded locally by full thickness dermal incisions above the mammary fat pads or remotely above the scapula 9 days later. Local, but not remote, wounding increased tumor size when compared with sham treatment. Injection of wound fluid close to the tumor site increased tumor growth, whereas in vitro wound fluid compared with serum increased the proliferation rate of 4T1 cells. Our results show that wound stroma can unfavorably influence growth of nearby tumors. This effect is T cell-dependent, as local wounding had no effect on tumor growth in nu/nu mice. The effect of wounding on tumor growth can be mimicked by acellular wound fluid, suggesting that T cells secrete or mediate secretion of cytokines or growth factors that then accelerate tumor growth. Here, we define an experimental model of wound-promoted tumor growth that will enable us to identify mechanisms and therapeutic targets to reduce the negative effect of tissue repair on residual tumors.

Laparoscopy-assisted distal gastrectomy for gastric cancer.

OBJECTIVE: The purpose of this study was to evaluate the safety and value of laparoscopy-assisted distal gastrectomy (LADG) for early stage gastric cancer (stages IA, IB, and II). MATERIALS AND METHODS: We retrospectively assessed 101 cases treated by LADG and compared to 49 contemporaneous cases treated by open distal gastrectomy (DG) between 2001 and 2006. Clinical variables, such as tumor diameter, operation time, blood loss, number of lymph nodes dissected, and length of stay were investigated. RESULTS: Tumor size (mm) was significantly smaller in the LADG group (p < 0.0001). Although operation time (min) in the two groups was similar (278 +/- 57 vs. 268 +/- 55), mean blood loss was significantly higher in the DG group (139 +/- 181 vs. 460 +/- 301, p < 0.0001). Fewer lymph nodes were harvested in the LADG group (27 +/- 14 vs. 34 +/- 19, p = 0.012). Hospital stay was longer in the DG group (13.3 +/- 8.5 vs. 16.7 +/- 10.5, p = 0.034). There was no mortality in either group. Postoperative surgical complications occurred in six (6%) of the LADG and four (8%) of the DG. CONCLUSIONS: The authors conclude that laparoscopy-assisted distal gastrectomy is a safe and useful operation for early-stage gastric cancers. If patients are selected properly, laparoscopy-assisted distal gastrectomy can be a curative and minimally invasive treatment for gastric cancer.

Exogenous pro-angiogenic stimuli cannot prevent physiologic vessel regression.

BACKGROUND: In healing wounds, rising levels of vascular endothelial growth factor (VEGF) induce a period of robust angiogenesis. The levels of pro-angiogenic factors in the wound begin to decline just before a period of vascular regression, suggesting that these mediators are necessary to sustain vessel density. The purpose of this study was to determine if the maintenance of pro-angiogenic stimuli in the wound would prevent physiological vessel regression. MATERIALS AND METHODS: A standard subcutaneous sponge wound model was modified by the addition of a mini-osmotic pump, allowing manipulation of the wound milieu by the addition of exogenous growth factors. After initial characterization of this model, exogenous VEGF (10 microg/mL), FGF (10 microg/mL), PDGF (10 microg/mL), or VEGF (10 microg/mL) plus FGF (10 microg/mL) were delivered to wounds and blood vessel density analyzed by immunohistochemistry. RESULTS: VEGF administration resulted in a transient increase in wound vessel density (P < 0.05). None of the pro-angiogenic growth factors (VEGF, FGF, PDGF, VEGF/FGF) were able to prevent vascular regression (P = NS). CONCLUSIONS: These findings suggest that the anti-angiogenic signals that mediate physiological vascular regression in wounds are strongly dominant over pro-angiogenic stimuli during the later phases of wound healing. Clinical manipulation of anti-angiogenic signals in addition to the currently used pro-angiogenic targets may be needed to achieve therapeutic modulation of blood vessel density.

Nutrition and wound healing.

The relationship between nutrition and wound healing--after injury or surgical intervention--has been recognized for centuries. There is no doubt that adequate carbohydrate, fat, and protein intake is required for healing to take place, but research in the laboratory has suggested that other specific nutritional interventions can have significant beneficial effects on wound healing. Successful translation into the clinical arena, however, has been rare. A review of normal metabolism as it relates to wound healing in normoglycemic and diabetic individuals is presented. This is followed by an assessment of the current literature and the data that support and refute the use of specialized nutritional support in postoperative and wounded patients. The experimental evidence for the use of arginine, glutamine, vitamins, and micronutrient supplementation is described. Most of the experimental evidence in the field supporting the use of specialized nutritional support has not been borne out by clinical investigation. A summary of the clinical implications of the data is presented, with the acknowledgment that each patient's plan of care must be individualized to optimize the relationship between nutrition and wound healing.

Adenoviral gene transfer of an NF-kappaB super-repressor increases collagen deposition in rodent cutaneous wound healing.

BACKGROUND: The transcription factor nuclear factor-kappaB (NF-kappaB) plays an essential role in inflammation. To date, no studies have investigated the effect of inhibiting NF-kappaB-mediated inflammation on normal cutaneous wound healing. We tested this by locally administering an adenovirus recombinant that constitutively expresses a super-repressor isoform of inhibitory-kappaB (IkappaB) into rats undergoing a well-established model of dorsal wound healing. METHODS: Seventy-two Sprague-Dawley rats underwent insertion of a sponge-pump construct into a dorsal subcutaneous pocket. One group of rats received pumps filled with the adenovirus expressing I-kappaB (rAd-Ikappab), a second group received pumps filled with adenovirus expressing green fluorescent protein (GFP) (rAd-gfp), and a third received pumps filled with normal saline (NS). Rats were killed in groups of 6 on days 1, 3, 5 and 7 postoperation. The wound fluid was analyzed for nitrite/nitrate (NOx) and tumor necrosis factor-alpha (TNF-alpha) concentrations. The wound fluid was assayed for hydroxyproline (OHP) content, an index of reparative collagen deposition. RESULTS: Administration of rAd-Ikappab for 7 days resulted in higher collagen deposition (OHP) compared with the rAd-gfp and NS groups. NOx levels were significantly higher in the rAd-gfp group on day 1 and marginally so on day 5. TNF-alpha quantitation analysis found no significant difference among the 3 groups. CONCLUSION: IkappaB expression through an adenoviral vector in the cutaneous wound may improve rodent healing, as shown by increased collagen deposition, through decreased inflammation. This mechanism appears to be TNF-alpha independent. Inhibition of NF-kappaB may reduce inflammation by reducing the local NOx concentrations.

Postoperative changes in body composition after gastrectomy.

Nutritional status is one of the most important clinical determinants of outcome after gastrectomy. The aim of this study was to compare changes in the body composition of patients undergoing laparoscopy-assisted gastrectomy (LAG), distal gastrectomy (DG), or total gastrectomy (TG). Total body protein and fat mass were measured by performing a multifrequency bioelectrical impedance analysis using an inBody II machine (Biospace, Tokyo, Japan) in 108 patients (72 men, 36 women) who had undergone LAG (n=24), DG (n=39), or TG (n=45). Changes between the preoperative data and results obtained on postoperative day 14 and 6 months after surgery were then evaluated. The mean preoperative body weight of the subjects was 57.6+/-10.7 kg, the mean body mass index was 22.5+/-3.4 kg/m(2), and the mean fat % was 24%+/-7%. In the immediate postoperative period (14 days), the body weight loss in the LAG group was significantly lower than in the DG and TG groups (2.5+/-0.9 kg vs. 3.5+/-1.8 kg and 4.0+/-1.9 kg, respectively; P < 0.0001). The body composition studies demonstrated a loss of total body protein rather than fat mass. Six months after surgery, body weight was not significantly different from preoperative values in the LAG and DG groups (-1.2+/-3.8 kg and -1.8+/-4.7 kg, respectively), but had decreased by 8.9+/-4.9 kg in the TG group (P=0.0003). A body composition analysis revealed a loss of fat mass in the DG and TG groups. The patients who underwent gastrectomy lost body protein mass during the early postoperative period. The type and extent of surgery has an effect on long-term body mass and composition. Bioelectric impedance analysis can be used to assess body composition and may be useful for nutritional assessment in patients who have undergone gastrectomy.

Cellular and physiological effects of arginine.

Arginine is a semi-essential amino acid that is required during periods of maximal growth, severe stress, and injury. Arginine is a substrate for protein synthesis but also modulates cellular biochemical functions via conversion to a number of biologically active compounds. Arginine is utilized by a vast variety of metabolic pathways that produce a variety of biologically active compounds such as nitric oxide, creatine phosphate, agmatine, polyamines, ornithine, and citrulline. Arginine supply is primarily regulated by two enzyme systems: arginase (part of the urea cycle) and nitric oxide synthase. Arginine has many effects in the body that include modulation of immune function, wound healing, hormone secretion, vascular tone, insulin sensitivity, and endothelial function. Arginine mediates its effects via nitric oxide independent and dependent pathways. Nitric oxide modulates many cellular functions that include vascular tone, expression of adhesion molecules, leukocyte adhesion, and platelet aggregation. Arginine modulates the development of atherosclerotic cardiovascular disease, improves immune function in healthy and ill patients, stimulates wound healing in healthy and ill patients, and modulates carcinogenesis and tumor growth. Thus, arginine is a biologically active dietary compound with numerous physiologic and pharmacological activities.