bluuebunny/crossref_metadata_2025 huggingface dataset:
104 Effect of Connective Tissue Growth Factor on Protein Kinase Expression and Activity in Human Corneal Fibroblasts
Authors: Radhakrishnan S.S. , Blalock T.D. , Schultz G.S.
Published: March 2007

Purpose: To evaluate expression patterns of protein kinases and protein kinase activities in human corneal fibroblasts treated with connective tissue growth factor (CTGF).

Methods: Human corneal fibroblast cultures were grown to confluence and treated with CTGF for 0, 5, and 15 minutes. Cytoplasmic protein extracts were obtained; protein kinase expression and activity arrays were performed using Kinetworks™ analysis (screening for expression of 75 different protein kinases and 31 different phosphoproteins). Further studies using extended time courses of CTGF exposure in corneal fibroblasts (0, 1, 2, 3, 4, 5, 10, 15, 30, and 60 minutes) were performed using immunoblot analysis to detect expression of protein kinase A catalytic subunit (PKA‐cat), and focal adhesion kinase (FAK). All results and were normalized by comparison to beta‐actin.

Results: After 5 minutes of exposure to CTGF, levels of active proteins increased for 21 of the 75 kinases analyzed. Some notable protein kinases that were induced include: death‐associated kinase 1, focal adhesion kinase (FAK), G‐protein coupled receptor kinase 2, protein kinase A catalytic subunit (PKA‐cat), protein kinase B alpha, and protein kinase C (ε, μ, and ζ, subunits). Extended time course analysis of PKA‐cat and FAK showed statistically significant increases in expression following CTGF stimulation within 15 minutes.

Conclusion: CTGF increased the levels of active protein kinases in human corneal fibroblast cultures, including PKA‐cat and FAK after 5 minutes of exposure. These results further our understanding of the signal transduction mechanism activated by CTGF in corneal fibroblasts. This suggests that CTGF mediates the effects of transforming growth factor beta (TGF‐β) on protein kinase expression and phosphorylation of second messengers in processes such as cell proliferation and collagen synthesis.

Acknowledgments: Funding source: NEI 05587
111 HB‐50: A Pre‐Clinical Study of a Prophylactic for Wound Infection
Authors: Harris Scott M , Zhang Lijuan , Parente Jody , Rodeheaver George T. , Falla Timothy J.
Published: March 2007

Topical prophylaxis against wound infection by an agent that is active against multi‐resistant bacteria does not generate resistance and is rapidly cidal would be of great clinical benefit. Peptides of the innate immune system have long been known to protect a wide range of organisms from attack by bacterial and fungal pathogens. Helix BioMedix Inc. has developed a short bioactive peptide antimicrobial modeled after these peptides. HB‐50 is an amphipathic cationic alpha‐helical peptide that has broad spectrum activity and is rapidly microbicidal. These attributes make HB‐50 an ideal candidate for wound infection prophylaxis. In vitro studies have demonstrated HB‐50 to be active against both gram‐positive and gram‐negative bacteria killing 5–7 log orders of bacteria within minutes. In addition, this peptide has potent activity against Vancomycin and Mupirocin resistant S. aureus. Preliminary testing of the peptide in a rat abraded skin infection model has shown the peptide’s effectiveness in preventing wound infection while not inhibiting wound healing. Additionally, the HB‐50 sequence has been specifically developed to be cost effective to manufacture and therefore is well suited for use as a topical antimicrobial agent.
112 Effects of High Voltage Pulsed Current on Bacterial Viability: An in Vitro Study
Authors: Kuykendall Cassie A. , Hutchinson Trish L. , Bloedaum Alan P.
Published: March 2007

Background and Purpose: High voltage pulsed current (HVPC) has been shown to have positive effects on wound healing due in part to proposed bacteriocidal effects in wound beds. The purpose of this study was to examine the effects of electrical stimulation (HVPC) using in vitro Streptococcus A and clinically‐accepted wound care guidelines.  Methods: Commercially available Streptococcus A was grown overnight in Trypticase Soy Broth. Standard 100 millimeter (mm) Petri dishes of Trypticase Soy Agar with 5% sheep blood were then streaked using sterile cotton swabs. 52 plates were subjected to 45‐minutes of HVPC with the following parameters: 120 pps, 200 V, 10‐second on/off cycle, and 2‐second ramp time. Negative polarity was used as the active electrode. 48 control plates were prepared identically without the application of the HVPC. Zones of inhibition were measured the following day at the positive and negative electrode sites to the nearest millimeter after a 24‐hour incubation period using the Kirby‐Bauer method. Results: Differences were calculated using a two‐way ANOVA with SPSS 11.5. Mean inhibition at the positive and negative electrodes was 0.00 mm ± 0.00 and 10.13 ± 4.13 mm, respectively. Significant between group effects were found for the control and experimental groups (F1,199 = 289.4, p < 0.001), negative and positive electrodes (F1,199 = 289.4, p < 0.001), and interaction of groups by electrodes (F1,199 = 289, p < 0.001).  Conclusion: In vitro, negative polarity HVPC using clinically accepted and feasible parameters can inhibit Streptococcus A growth around the negative (active) electrode. No growth inhibition was noted around the positive (dispersive) electrode. This could provide an alternate means of infection control in assistance with wound healing.
113 Does Wound Care Education Speed Healing of Infected, Acute Wounds?
Authors: Sherman J , Lyndon A , Hopf H
Published: March 2007

Background: Wound infections cause significant morbidity in women after C‐Sections. Regular lectures on wound care were instituted for obstetrical residents at our institution in 1999. We hypothesized that resident education would decrease time to complete closure in post‐C‐Section wounds healing by secondary intention.
Methods: With IRB approval, a retrospective chart review was performed to obtain the following information in patients treated pre (1996–1999) versus post (2000–2003) education intervention: age, race, occupation, social support, medical co‐morbidities, medications, week of gestation at delivery, duration of 1st stage of labor, length of ruptured membranes, meconium, chorioamnionitis, Group B Strep status, indications for C‐Section, antibiotics, wound treatment, wound culture, wound dressing, time to full closure, number of visits for wound care (home, office), and need for repeat procedures.
Results: Data are currently being analyzed.
Conclusions: If resident education reduces the time to full closure, then we will evaluate expanding this teaching approach to other departments using a randomized prospective study.
114 A Comparison of in vitro Disc Diffusion and Time‐Kill Kinetic Assays for the Evaluation of Antimicrobial Activity of Silver Containing Wound Dressings
Authors: Gallant Corrie L. , Yin Hua Q. , Langford Rita E. , Olson Merle E. , Hart David A. , Burrell Robert E.
Published: March 2007

There is a plethora of new silver containing dressings on the market today. Various manufacturers attempt to show that their dressings are the most efficacious and therefore should be preferentially employed by health care workers based on the results of their in vitro tests. However, there have been no studies to date that clearly identify which in vitro tests are appropriate for comparison purposes. The available literature suggests that there are problems with disc sensitivity assays in terms of assessing the antimicrobial efficacy of silver. Spadaro (1985) has shown that the diameters of zones of inhibition were not proportional to the concentration of anodic silver in complex test media. Further, Richards et al (2001) have shown that zones of inhibition did not appear to correlate to log reduction assay data for silver containing dressings. Other authors favour log reduction or time kill assays as they better compare to clinical data. The purpose of this study was to determine which in vivo test is most appropriate for evaluating the antimicrobial efficacy of silver‐containing dressings. This was done by testing 8 different silver containing dressings and 2 non‐silver (mafenide acetate or hexamethylene biguanide) containing topical agents against 17 clinically relevant microorganisms using both zone of inhibition assays and time‐kill kinetic assays in complex media. The results for the two assays were then correlated to determine if the methods generated similar results. It was determined that the two methods do not correlate at all. This is most likely a result of the silver interacting with the media in the zone of inhibition test, thus invalidating the results of this test. We therefore conclude that zone of inhibition data generated for silver‐containing dressings is of little value when assessing antimicrobial efficacy, and that time‐kill assays are of greater use.
117 Care of the Thermally Injured Breast
Authors: Ortega R.E. , Riina L.H. , Davenport T.A. , Glickman L.T. , Simpson R.L. , Burger K.M. , Hangan D.
Published: March 2007

Introduction: Thermal injury to the female breast is one of the most challenging aspects of aesthetic burn surgery today. As the ability to provide coverage for large body surface injuries has progressed greatly in recent years; attention can now be directed towards achieving aesthetically pleasing results. Breast reconstruction in the burn patient can be accomplished in several ways. In planning the reconstruction, one must account for gender, age, and stage of breast development at the time of injury. The following is the treatment protocol utilized at our institution. After determining the level of injury the injured area is cleansed and treated with topical anti‐microbial agents such as Silvadene. If the decision is made to excise and graft the injured area one of two algorithms is followed. The first choice involves excising the burn and placing a split thickness skin graft to the area involved. This is done by placing a sheet graft and using aerosolized fibrin sealant to affix it to the wound bed. If the burn involves deeper elements of tissue then a second approach is taken which includes excision of the burn down to the level of fascia with preservation of the breast mounds and the nipple areola complex (NAR). The (NAR) is spared excision and allowed to heal. Reconstruction of the (NAR) can be deferred for a secondary procedure depending upon the response to primary healing. A split thickness skin graft is then applied to the area of injury. Again a sheet graft is preferred and fibrin sealant is utilized to improve graft fixation and contour. We attribute our excellent results to the sheet grafts and fibrin sealant used. It should be noted that the increased vascularity of the breast fat when compared to fat located elsewhere in the body allows the grafts to adhere and survive on this generally difficult to graft surface.

Methods: We identified five female patients at our institution over the last 18 months with thermal injuries to the breasts. Each patient was placed into one of the two treatment algorithms.

Results: The five patients had excellent outcomes. Breast mounds and symmetry were preserved. Further development of the breast was allowed in each patient. One patient even underwent a breast augmentation after surviving a 50% TBSA injury. Proper use of fibrin sealant and sheet grafts account for the excellent results seen at this institution.

Conclusion: Following careful evaluation of the burned female breast cosmetically and functionally acceptable results can be attained when following our institution’s protocol for breast reconstruction in the female burn victim.
126 Scrotal and Penile Reconstruction using VAC Therapy
Authors: Snyder N , Gould LJ
Published: March 2007

Fournier gangrene is a rapidly progressive necrotizing soft tissue infection that often leaves patients with a large integumentary deficit to their penis and scrotum. Treatment of the skin defect has been described in a multitude of ways including split thickness skin grafting, burying the testes in the thighs, thigh flaps, and a variety of myocutaneous and fasciocutaneous flaps.

Many of the previously described techniques have worked well for smaller defects and for closure of wounds but fell short of aesthetic reconstruction of sexual organs. We describe a technique using split‐thickness skin grafts with suction dressings or bolster. This was employed using the VAC©(Vacuum Assisted Closure Device, KCI©). This technique creates a natural appearing scrotum that holds the testes away from the body in a physiologic manner and surfaces the penis with a seemingly natural appearance in a functional manner. The technique provides a nearly 100% graft take and greatly decreases hospital length of stay.

A case report demonstrates a patient’s full recovery to his professional, social, and sexual life within six weeks of initial reconstructive treatment. Our technique provides a reliable treatment with good functional and aesthetic outcomes to a difficult problem.
128 Wound Healing Morphology in an Acute Wound Healing Study
Authors: Serena T.
Published: March 2007

Background: Physiological healing in acute wounds generally occurs by granulation from the wound base and by migration of epithelium from the wound edges toward the center of the wound in a uniform, concentric fashion. The Wound Healing Cooperative Group (WHCG) compared this phenomenon in acute wounds treated with standard care versus acute wounds pharmacologically stimulated with growth factor therapy.

Methods: With informed consent, 20 normal healthy volunteers underwent four 6 mm biopsies of the flexor surface of both forearms. The biopsy sites were randomly assigned to a control arm (daily bacitracin) or to one of three treatment arms: i) rhPDGF‐BB 0.01% gel Q.D., ii) rhPDGF‐BB Q.O.D., iii) daily rhPDGF‐BB (Q.D. × 7 days followed by bacitracin alone daily). Wound morphology was carefully examined and photographed daily until complete healing was achieved.

Results: There were distinct differences in the morphological pattern of healing seen between control wounds and growth factor‐stimulated wounds. Acute wounds treated with bacitracin tended to heal in circumferential fashion, as predicted. Growth factor‐stimulated wounds, by contrast, exhibited accentuated angiogenesis (granulation), with non‐uniform epithelial islands streaming into the wound.

Conclusion: This pilot study suggests that rhPDGF‐BB influences acute wound healing by promoting accelerated granulation and epithelialization. Accelerated healing is manifest by different healing morphologies. The biological basis for these differences requires further histological and molecular analyses.

Acknowledgements:

Funding from The Angiogenesis Foundation.
132 Differential Regulation of Osteopontin and Matrix Metalloproteinases During Diabetic Wound Healing
Authors: Sharma A. , Singh A. K. , Warren J. , Maheshwari R. K
Published: March 2007

Delayed wound healing is a common complication of diabetes resulting in significant clinical morbidity. The diabetic wound exhibits impaired cellular infiltration and consequently inadequate granulation tissue formation. Additionally, altered patterns of apoptosis. In this study, we have evaluated the differential gene expression pattern in transgenic diabetic female mice, 5–6 weeks old, in a full thickness cutaneous punch wound model. We assessed the role of matrix metalloproteinases (MMPs) and differential gene expression pattern at 4th, 7th and 11th day post wounding. Supernatants obtained from diabetic wound tissue homogenates were subjected to zymogram analysis. The data showed that MMPs were expressed at higher level by 4th day post wounding, whereas expression of MMPs were down regulated towards the 11th day post wounding suggesting their role during early phase of wound healing.

The pathway specific gene array data demonstrated differential regulation of several growth factors, transcription factors and other related genes such as fibroblast growth factors and their receptors, ID3 and restin respectively. The cytokine/extracellular matrix protein osteopontin (OPN), an important component of cellular immunity and inflammation also showed higher expression after 4 days post wounding. The expression of OPN remained at higher level after 11 days post wounding in diabetic mice, whereas the expression were down regulated to basal level in normal wounded animal suggesting that the expression of OPN was concomitant with the extent of healing. Other adhesion molecules such as integrin αV and PECAM‐1 were also differentially regulated. Though a single gene may not be solely responsible for any defect or impairment in healing as it is a very tightly controlled and regulated process, however, a detailed study of these gene(s) may shed some light to the delayed healing in diabetic mice.

Acknowledgements: (These studies are supported by NIH Grant G174KT.)
135 Induction of Gene Expression for EGF‐like Ligands in Porcine Wounds Following Receptor Overexpession
Authors: Okwueze M.I. , Cardwell N.L. , Pollins A.C. , Nanney L.B.
Published: March 2007

A rationale to explain the role of the large and redundant family of EGF cytokines with multiple receptor combinations has proven elusive. We examined endogenous gene expression levels during wound repair for four EGF‐like cytokines (Transforming growth factor‐alpha (TGFα), Heparin‐binding epidermal growth factor (HB‐EGF), Epiregulin (EPR) and Betacellulin (BC) using a quantitative real‐time PCR technique. In addition, cytokine gene expression was evaluated after overexpression of different of EGF receptor forms (erbB‐1, erbB‐3 and erbB‐4) or a control B‐galactosidase gene using an adenoviral transfection strategy to introduce either homo or heterodimer receptor combinations. At baseline, endogenous gene expression levels of TGFα and HB‐EGF were 10‐fold and 100‐fold higher than EPR and BC, respectively. Expression of TGFα was increased by 3‐fold in the receptor combination of erbB‐1/erbB‐4 expressing wounds. Recent studies on the EGF‐related polypeptides HB‐EGF, EPR and BC have shown preferential binding to erbB‐4. Our study suggests that wounds predominantly expressing erbB‐4 showed a 2.5‐fold increase in gene expression of HB‐EGF. A synergistic 4‐fold increase in gene expression was noted when erbB‐4 overexpression was treated with HB‐EGF ligand. In contrast, wounds transfected with erbB‐4 alone showed no change in EPR gene expression compared to controls. However, wounds treated with TGFα showed 3‐fold increase in EPR gene expression and EPR potentiated itself. Similarly, wounds transfected with erbB‐4 and treated with growth factors TGFα and EGF upregulated the gene expression of BC by 4‐fold as compared to controls. Interestingly, wounds overexpressing erbB‐3 with topica application of EGF accentuated expression of TGFα by 7‐fold. In histologic sections, these wounds showed a 2‐fold increase in the dermal region compared to control. Thus, the EGF‐like family of ligands and its homo‐or heterodimer receptor pairs have the potential in vivo to evoke differing profiles or intensities of gene expression patterns as they mediate wound repair.
136 Celosia Argentea Linn. Leaf Extract Improves Wound Healing in Rat Burn Wound Model
Authors: Priya Kulasekaran S , Babu Mary , Wells Alan
Published: March 2007

Celosia argentea is used in traditional medicine for sores, ulcers, and skin eruptions. Therefore, we investigated the healing efficiency of an ointment formulated on a alcohol extract of Celosia argentea leaves (CA) in a rat burn wound model. Wound closure occurred earlier in the treated rats (15 days versus 30 in the untreated group; P < 0.05). Collagen and hexsoamine content of the granulation tissue increased at a faster rate in the treated wounds. To probe the cell biologic basis of this effect, we found that this extract promoted cell motility and proliferation of primary dermal fibroblasts but did not alter these responses in primary keratinocytes. In short, we demonstrate a salutary action of the Celosia argentea extract on wound healing and suggest that this may be due to mitogenic and motogenic promotion of dermal fibroblasts.
138 Molecular Pathogenesis of Chronic Wounds: The Role of β‐Catenin and C‐MYC
Authors: Stojadinovic O. , Brem H. , Lee B. , Vouthounis C. , Entero H. , Tomic‐Canic M.
Published: March 2007

The lack of understanding of molecular pathogenesis of impaired healing in chronic ulcers leads to a serious health issue that contributes to excessive limb amputations and mortality. Using biopsies from patients with chronic wounds, skin organ culture and primary keratinocytes in culture we identified that β‐catenin and its downstream target, c‐myc, play important role in development of chronic wounds. In contrast to normal epidermis, we observed significant nuclearization of β‐catenin and elevated c‐myc expression at the non‐healing wound edge of patients with chronic ulcers. In vitro studies indicated that activation and stabilization of nuclear β‐catenin inhibits wound healing and keratinocyte migration by: blocking EGF response and inducing c‐myc. Using Affymetrix large scale microarrays we found that β‐catenin downstream target, c‐myc, is induced in skin by an inhibitor of wound healing (glucocorticoids) and repressed in the initial phase of normal wound healing, 4 to 48 hr, whereas it becomes de‐repressed at 96 hr post wounding. Therefore, the activation of β‐catenin/c‐myc pathway(s) contributes to impaired healing by inhibiting of keratinocyte migration and altering keratinocyte differentiation. The presence of activated β‐catenin and c‐myc in the epidermis of chronic wounds may serve as molecular markers of impaired healing and future targets for therapeutic intervention. While β‐catenin signaling has been implicated in epithelial development and oncogenesis its role in wound healing has never been postulated. This further illustrates the importance of “tissue context” specificity, because β‐catenin in the context of malignant tissue promotes invasion whereas in the context of a wound environment does the opposite.
142 β‐Catenin and Carm‐1 as Co‐Repressors of Glucocorticoid Receptor Lead to Inhibition of Keratinocyte Migration
Authors: Stojadinovic O. , Vouthounis C. , Lee B. , Stallcup M. , Tomic‐Canic M.
Published: March 2007

Ability of keratinocytes to migrate relays on appropriate citoskeletal network and is one of the key prerequisites of wound healing. Keratins K6 and K16 are the cytoskeletal components that mark migrating keratinocytes. We found K6/K16 to be differentially regulated in microarray experiments: induced in the wound healing and suppressed by inhibitors of wound healing, glucocorticoids (GC). We have shown previously that GC suppress K6/K16 expression through a unique molecular mechanism that involves four monomers of glucocorticoid receptor (GR). Because β‐catenin interacts with members of the nuclear receptor family, such as retinoic acid receptors (RAR) and androgen receptor (AR) and participates in both activation and repression of transcription and similarly to GC inhibits keratinocytes migration we hypothesized that β‐catenin participates in GC‐mediated repression of K6/K16. To test if β‐catenin participates in K6/K16 suppression by GC we used co‐transfection experiments, with primary human keratinocytes. By itself, β‐catenin did not affect K6/K16 expression. However, in the presence of GC, β‐catenin acted as a GR co‐repressor further suppressing K6/K16 expression. Moreover, protein arginine methyltransferase CARM‐1 enhances this co‐repression, suggesting that the complex that suppresses K6/K16 promoters contains GR, β‐catenin and CARM‐1. Similarly, β‐catenin functioned as a co‐repressor of GR even if we just incubated keratinocytes with LiCl (stabilized β‐catenin) rather than transfected its expression plasmid. It has been shown that β‐catenin and CARM‐1 bind each other in a co‐activator complex with AR, but it was not known if this complex has a co‐repressing capacity. We have shown that by participating in the GC‐mdiated repression of K6/K16 transcription as a co‐repressor with CARM1, β‐catenin contributes to the inhibition of keratinocyte migration through altering the cytoskeletal network and subsequent inhibition of wound healing.
144 The Upregulation of Wound Healing Related Genes in Venous Ulcerations Treated with Apligraf, a Possible Mechanism of Action
Authors: Serena T.E. , Finley C.
Published: March 2007

Background: Apligraf, a bioengineered living skin construct, has been shown to accelerate the healing of chronic venous leg and diabetic foot ulcerations. However, to date the mechanism of action of Apligraf in the wound healing process is not well understood.

Objective: The primary objective of this study was to determine the levels of expression of selected wound healing related genes in venous leg ulcers treated with Apligraf in comparison to ulcers treated with standard multi‐layer compression therapy alone. Gene chip technology was employed.

Methods: Three patients were randomized into the Apligraf or standard treatment arms. A baseline 6 mm punch biopsy was obtained prior to the initial application of Apligraf or multi‐layer compression therapy. A second biopsy was obtained depending on the randomly chosen biopsy schedule (weeks 1, 2 or 4 following Apligraf application or initiation of compression). The biopsy specimens were snap frozen and later analyzed using microarray gene chip technology.

Results: The patients treated with Apligraf demonstrated an up‐regulation of the genes thought to be important in the wound healing process.

Conclusion: The results from this pilot study suggest that Apligraf may function by an up‐regulation of the genes involved in wound healing as opposed to compression therapy which works primarily in a mechanical fashion. Based on these results further study is needed.
147 Persistence and Expansion of Seeded Marrow Cells in Dermal Substrate
Authors: Liu PY , Wang XT , Ponte C , Tang JB , Badiavas EV
Published: March 2007

Bone marrow cells have been used to potentiate wound healing in chronic wounds (Badiavas and Falanga, 2003). We postulate that the performance of these autografted cells may be enhanced by seeding them into scaffolding with normal skin architecture; indeed that the differentiation of the precursor cells may be determined by the juxtaposition of intact extracellular matrix. This might in turn lead to a prefabricated skin substitute capable of recapitulating fully functional skin.

We are seeding murine gfp+ bone marrow cells, containing mixed mesenchymal and hematopoietic precursor cells into Alloderm®, a decellularized preserved skin graft. This seeding is potentiated by meshing the skin and gentle centrifugation of fresh marrow into the dermal aspect of the allograft. Other samples were merely co‐cultured with the cells in Dexter’s media. Duplicate samples were fixed and stained with DAPI at 1, 3, 7, and 14 days. Viewed under fluorescence, gfp+ cells are bright green.Comparing these images with DAPI‐staining allows viable cells to be identified which are gfp+. H&E micrographs were used to ascertain cellular morphology.

Results: Scant cells were observed in the allografts at the early time points. The cells were pleiomorphic with large nuclei, resembling hematopoietic precursors(Fig 1). Later, at days 7 and 14, more abundant cells were apparent, including some in the interior of the dermis. This signifies either migration or seeding via centrifugation. The morphology of these cells is significantly different; the spindle shape suggests a fibroblast (Fig 2). In addition, the population of gfp+ cells has markedly expanded by day 14, attaining near confluence in some sections (Fig 3). No cells were seen in controls (Fig 4).

Conclusion: True skin replacement will only be possible when the panoply of skin appendages can be recapitulated. The experiments presented here are evidence that pluripotential precursor cells can be seeded into dermal matrices providing an optimal environment for regeneration.

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148 2‐Methoxyestradiol, an Endogenous Estrogen Metabolite, Regulates Apoptosis in Skin Microvascular Endothelial Cells
Authors: Philip A. , Horvat A‐M , Chowdhury M.
Published: March 2007

Programmed cell death or apoptosis regulates the sequence of cellular events that are involved in the wound healing process. Understanding the mechanisms controlling apoptosis during tissue repair may lead to the development of modalities to improve healing and reduce scarring. 2‐Methoxyestradiol (2ME), a naturally occurring metabolite of 17‐ estradiol, has recently emerged as a promising anti‐proliferative and angiostatic agent with minimal toxicity at pharmacological doses. However, the mechanism by which 2ME induces its anti‐proliferative and apoptotic activities is uncertain, and its physiological function remains to be determined. In the present study we examined the effects of physiological and pharmacological concentrations 2ME in human skin microvascular endothelial cells.

Our findings show that 2ME rapidly activates JNK and p38 MAP kinase pathways at both physiological and supraphysiological levels. Our results indicate that the rapid activation of p38 and JNK are not required for 2ME‐induced inhibition of DNA synthesis, caspase‐3 activation, or cell cycle arrest, and thus apoptosis, in microvascular endothelial cells. On the contrary, our results suggest that rapid activation of JNK and p38 MAP kinases likely mediates an early pro‐survival response to 2ME. The fine regulation of survival versus apoptotic signals may contribute to the ability of 2ME to elicit its effects in endothelial cells. Since endothelial cells play an essential role during wound healing, identification 2ME as a potent regulator of endothelial cell proliferation and function warrants further studies of its therapeutic potential as an agent to manipulate wound repair events.
150 A Study on Scientific Validation of the Indian Traditional Wound Healing Agent
Authors: Gnanamani A. , Gopal R. , Babu Mary , Sadulla S.
Published: March 2007

Maththaan thailam, the Datura leaf juice is traditionally used for healing of wounds and related ailments in India. However, the scientific validity of the thailam has not been explored in the literatures. In the present study, the efficacy of maththaan thailam on healing of three types of wounds namely Open, Burn and Diabetic wound in animal models has been studied. Healing pattern and biochemical characterization of granulation tissue were assessed over the healing periods. Studies were more concentrated on to evaluate the change in the matrix metallo proteinase profile in the three different types of wound with respect to the applied thailam. Further supports were also expected from the antioxidant levels and histopathological examination of granulation tissue. It has been observed that among the three types of wounds, delayed healing was observed with diabetic wound. Complete healing of 2 × 2 cm2 wound size was achieved only after 22 days, whereas burn and open wound took only 16 and 9 days for complete closure. Further, MMP’s profile also showed a variation in the expression of MMP2 and MMP9 with respect to healing property. Biochemical and histochemical results and the antioxidant levels are highly correlated with the healing property irrespective of the wounds
006 Development of a Novel Matrix Metalloproteinase‐Inhibiting Wound Dressing
Authors: Skarja Gary A. , Gorbet Maud B. , Lawson‐Smith Rebecca K. , May Michael H. , Sefton Michael V.
Published: March 2007

Rimon Therapeutics develops synthetic therapeutic polymers (Theramers™) that combine bioactivity with a broad range of desirable mechanical and physical properties. The MI Theramer™ is a polymer that modulates the activity of matrix metalloproteinases (MMPs).. Excessive proteolytic cleavage of extracellular matrix and growth factors resulting from elevated levels of MMPs is believed to contribute to the impaired wound healing associated with chronic, non‐healing wounds. MI Theramer™ beads suspended in ThermaGel™(a thermoreversible gel) make up MI‐Gel™ Dressing. The dressing may be applied as a liquid that is poured into a wound, where it will gel on contact. The dressing can be reliquified and removed easily without re‐injuring the wound by cooling it.

In vitro inhibition of collagenase type IV (equivalent to MMP‐2) from clostridium histolyticum (Molecular Probes) was measured. A dose‐dependent inhibition of MMP‐2 activity was measured for MI Theramer™ beads alone and suspended in ThermaGel™ indicating that MMP‐2 is able to diffuse into the gel and bind to the beads suspended therein. In vivo efficacy was demonstrated by observing the degradation of glutaraldehyde cross‐linked gelatin tubes using a mouse subcutaneous pouch model. Gelatin zymography on extracts collected from the gelatin tubes co‐implanted with MI Theramer™ beads also showed reduced MMP‐2 and MMP‐9 activity in comparison to controls.

The insoluble MI Theramer™ beads were able to modulate MMP activity both in vitro and in vivo. The beads retained their inhibitory activity when they were suspended in a novel thermoreversible gel (ThermaGel™). The combined MI‐Gel™ Dressing may have promise as a novel chronic wound healing product.
007 Rebalancing Wound Biochemistry Improves Healing: A Clinical Study Examining Effect of Promogran
Authors: Cullen B. , Kemp L. , Essler A. , Wallenfang‐Sohle K. , Stadler R.
Published: March 2007

Normal wound healing is a carefully controlled balance of new tissue formation and destructive processes necessary to remove damaged tissue. Within this complex environment there are many points of regulation, which control the biological processes necessary to achieve wound repair. An alteration in any of these processes can result in an imbalance of the biochemical components, which ultimately results in delayed wound closure and the formation of a chronic wound. Therefore, we propose that an effective therapeutic approach would modify this hostile environment and redress this imbalance.

In this study we have evaluated the effect of PROMOGRAN in vivo, in patients with venous leg ulcers. Wound fluid samples were collected and analyzed from a number of patients prior to and during treatment with PROMOGRAN.

Our results have shown that wounds, which respond to PROMOGRAN treatment, have also shown a decrease in protease activity in the corresponding wound fluid samples. Whilst it is impossible to determine whether this reduction in protease levels is responsible for healing or merely symptomatic of other changes occurring within the wound, we have shown that a decrease in proteolytic activity is concomitant with healing.

This study provides clinical evidence that PROMOGRAN can rebalance the chronic wound environment in situ and thereby promotes wound repair.
009 Evaluation of an Ozone‐Based Wound Management System
Authors: Denvir A.J. , Blankenburg A.C. , Holder I.A.
Published: March 2007

The onset of infection can lead rapidly to sepsis, septic shock, and eventually death. Considering the high costs of hospitalization and the added trauma and discomfort to the patient, improved methods for infection control are needed. Ozone offers a specific solution to the problem of effective management of microbial wound contamination. Despite the advantages, several technical barriers have prevented ozone‐based disinfection treatments from becoming more widely investigated. Negative press from years of unsubstantiated medical successes has made the medical community wary of even legitimate ozone technologies. Because there has been no research into using ozone in infection prevention, much of the hardware has not been developed to the stage where the technology can be used in clinical trials. Lynntech Inc., over the past 5 years, has been working to develop a pre‐clinical ozone‐based wound management system. This new hardware uses an electrochemical ozone generator that can deliver a predetermined quantity of ozone gas to a specific wound site. Electrochemically generated ozone has been shown to be effective against a range of gram negative and gram positive bacteria and our studies with various mimetic wound systems have shown that with controlled delivery, ozone can be utilized as either a disinfectant or as a biostat. The device is well suited for hospital use as it operates of low voltage power supplies and unlike other ozone generation technologies will not interfere with other electronic equipment while it is in operation. This paper outlines both the advantages and limitations of using an ozone based system as a wound management tool and looks to the future research that needs to be performed to substantiated the initial research findings.
010 Structural Characterization of Small Intestinal Submucosa (SIS) using Cryo and Traditional Scanning Electron Microscopy (SEM)
Authors: Janis A.D. , Sherman D.M.
Published: March 2007

Composition and structure determine the fate of biomaterials in the body, encouraging or discouraging acceptance, cellular infiltration, and degradation or incorporation into host tissue. Most biomaterials research has been focused on composition, while structure remains a relatively uninvestigated factor in biomaterial success. Naturally derived biomaterials may provide optimal structure for cellular repopulation in certain applications. Porcine small intestinal submucosa (SIS) is an acellular, naturally occurring extracellular matrix used in the clinical treatment of soft tissue injury, including pressure, venous, and diabetic ulcers. Numerous studies have demonstrated that SIS induces host tissue infiltration, tissue‐specific remodeling, and repair. SIS consists primarily of collagen, although the presence of other proteins, growth factors, glycosaminoglycans, and proteoglycans has been reported. SEM was used to characterize the cellular environment provided by this biomaterial. Cryo‐SEM was used to image mechanically isolated, oxidatively disinfected, lyophilized, sterilized, and rehydrated SIS. The fractured cross‐sectional faces of intact intestine, oxidatively disinfected SIS, and lyophilized SIS were imaged with traditional SEM. Comparison of the images utilizing cryo and traditional preparation techniques show that SIS is a heterogeneous matrix consisting of textured fibers of mixed size. There are differences in the structure of the mucosal and serosal surfaces. Although the matrix architecture shows signs of collapse and delamination following lyophilization, much of the native structure is preserved. These changes are preserved following rehydration. The effects of these structural changes on cellular reaction and infiltration are unknown, but the naturally complex architecture‐clearly facilitates host interaction on thecellular and subcellular levels. This work was supported by Cook Biotech Incorporated.
022 Warming Surgical Wounds: Effects on Healing and Wound Complications
Authors: Whitney J.D. , Dellinger E. P. , Wickline M.
Published: March 2007

Aims: Compare postoperative local warming to standard incision care on: 1) healing response and 2) incidence of wound complications.

Methods: This two group randomized pilot study (N = 54) tested the wound healing effects of post‐operative warming in patients having gastric bypass or colectomy procedures. Patients randomized to warming received their first 1‐hour treatment in the post anesthesia recovery unit followed by 5 additional treatments in the first 48 hours after surgery. Healing response was evaluated by hydroxyproline and mRNA for pro α 1(I) collagen content of a subcutaneous ePTFE implant removed on postoperative day 9. Patients were followed for the first two postoperative clinic visits (6–8 weeks). CDC Surgical Site Infection Criteria were used to evaluate wound infection and complications.

Results: Six warmed (22%) and 10 standard care (37%) patients experienced healing problems.

Incidence of Wound Infection by CDC classification.

  Incidence of Wound Infection by CDC classification.  

Group
Superficial
Deep Incisional
Organ Space

Warming
2
3
1

Standard Care
4
4
2

Mean hydroxyproline levels did not differ, though there was a trend toward higher amounts in warmed patients (332 ± 185 ng/cm ePTFE vs. 286 ± 165 ng/cm). Differences in mRNA for pro α 1(I) collagen approached significance (Chi‐Square 5.4, p = .06) with warmed patients showing more cells with abundant positive message.

Conclusions. Clinical and cellular outcomes suggest local incision warming may benefit healing by reducing infection and influencing fibroblast collagen production.
011 Inflammation‐induced Angiogenesis: Potential Roles of IL‐8. and VEGF
Authors: Yao M. , Dueck M. , Martins‐Green M.
Published: March 2007

Inflammation is invariably accompanied by angiogenesis; both are important in wound healing. However, it is not known how angiogenic factors that are present during the inflammatory phase of wound healing are involved in inflammation‐induced angiogenesis. In this study, we address the contribution of two such factors, interleukin‐8 (IL‐8/CXCL8) and VEGF, in inflammation‐induced angiogenesis. We determined the relative levels of these molecules during the first two weeks of the wound healing process in partial‐thickness skin wounds of rabbits. IL‐8 levels increase rapidly after wounding, reaching maximal levels after 24 hr; in contrast, VEGF levels peak at 3 days. We also determined the time course of the infiltration of various inflammatory cells and production of new blood vessels after wounding. Neutrophils were markedly increased 4 hr after wounding, and their level peaked at 24 hr. Monocyte infiltration peaked 48 hr after wounding and decreased by day 3. The number of blood vessels was significantly increased by day 3 after wounding and continued to increase through days 5 and 7.

In order to determine the contribution of IL‐8 and VEGF to the angiogenic process, we performed the wounding experiments in the presence and absence of IL‐8 and VEGF antibodies. Treatment with IL‐8 antibodies blocked the early stage of blood vessel formation but did not alter the late stage; in contrast, VEGF antibodies blocked the late stage without altering the early stage. Currently, we are performing studies using inhibitors against IL‐8 and VEGF receptors to determine whether we observe similar results; we are also beginning to decipher molecular mechanisms involved in this process. This study suggests that IL‐8 and VEGF play complementary roles in stimulating angiogenesis during wound healing.
017 Lower Expression of P‐Selectin may Modulate the Fetal Inflammatory Response:
Authors: Zhu X. , Cass D. , Smith C.W. , Olutoye O.O.
Published: March 2007

Fetal dermal wound healing is characterized by a minimal inflammatory response and absence of scar. In order to determine the factors contributing to the minimal inflammatory response in the fetus, the role of P‐selectin in the interaction and transmigration of leukocytes through vascular endothelium was investigated. Methods: Primary endothelial monolayers were established from adult porcine central veins and from umbilical veins of mid‐gestation porcine fetuses. The ability of the endothelial cells to capture and enable transmigration of adult leukocytes was studied under flow conditions of 4 dynes/sec, with and without prior stimulation of the endothelial cells with TNF‐α or IL‐1β. In addition, P‐selectin mRNA expression was determined by quantitative real time PCR. All data were analyzed by ANOVA.  Results: In response to IL‐1β 10 ng/ml stimulation, adult endothelial cells manifested a 10 fold increase in P‐selectin mRNA expression while fetal endothelial cells mounted only a 3.5 fold increase. 100 ng/ml was required to mount a 10 fold increase in fetal P‐selectin expression while no further increase in adult P‐selectin expression was noted at this dose. Leukocyte capture, demonstrated by rolling of neutrophils over the endothelial surface, was more efficient on adult monolayers compared to the fetus. This is inversely related to the rolling velocity which was significantly slower in the adult (5.3 ± 0.6 vs 12.4 ± 1.0 μm/mm2). Ultimately, the number of neutrophils transmigrating across adult endothelial monolayers under flow conditions was significantly higher compared to the fetal monolayer (199 ± 18 vs 72 ± 9 cells/mm2). Conclusion: Lower P‐selectin expression on fetal endothelial cells may account, in part, for the minimal inflammatory response noted following fetal dermal injury.
021 Contact Sensitivity in Patients with Leg Ulcerations: A North American Study
Authors: Saap L. , Fahim S. , Arsenault E. , Pratt M. , Pierscianowski T. , Falanga V. , Pedvis‐Leftick A.
Published: March 2007

Background: Over the last two decades, there have been a number of studies in Europe on contact sensitivity in patients with chronic leg ulcerations with a frequency of positive patch test results ranging from 40 to 82.5%. The prevalence of sensitization has not been studied in North America. Furthermore, many of the newer dressings and wound care products in the market have not been studied for contact sensitivity in patients with chronic wounds.

Objectives: 1) To determine the prevalence of allergen sensitivity in patients with history of leg ulcers in two North American study centers, 2) to compare our results to the European studies and to the North American Contact Dermatitis Group (NACDG) database and 3) to help delineate a standard battery of allergens for patch testing in North American leg ulcer patients

Methods: 54 patients with an active or past leg ulcer were prospectively entered in the study. The patients were patch tested to both the NACDG Standard series, as well as, a comprehensive supplemental series of 48 allergens including wound care medicaments and dressings.

Results: 63% of patients were sensitized to at last one allergen. The most common allergens were Balsam of Peru (29.6%), bacitracin (24.1%), fragrance mix (20.4%), wood tar mix (20.4%), propylene glycol (13.5%), neomycin sulfate (13%), benzalkonium chloride (13%), carba mix (11.1%), nickel sulfate (11.1%) and Duoderm CGF (11.1%). Duoderm CGF was the most allergenic dressing in our study group.

Conclusion: There is a high incidence of positive patch tests in patients with past or current leg ulcerations. Using a modified leg ulcer series along with the standard NACDG series is very important in evaluating patients with leg ulcers.
Critical influences affecting response to various treatments in young children with ADHD: a case series
Authors: Heriot S. A. , Evans I. M. , Foster T. M.
Published: March 2007

Background  While the use of stimulant medication as a treatment for children with attention deficit hyperactivity disorder (ADHD) has been the most studied therapy in child psychiatry, there is debate about its use with young children. This study describes a series of cases seen in a normal clinical context, treated with one of four different treatment programmes.

Methods  Sixteen pre‐school children diagnosed with ADHD and their parents were randomly assigned to receive one of four treatments: (1) 0.3 mg/kg methylphenidate, parent training programme; (2) 0.3 mg/kg methylphenidate, parent support programme; (3) placebo medication, parent training; and (4) placebo medication, parent support. Changes were assessed at the individual level, using clinical observations, parent and teacher rating scales and measures of parenting and family factors.

Results  Children were more likely to improve when the treatment involved at least one active component (medication or parent training). However, there was notable variability in individual parental and child participants' responses to all treatment conditions, indicating the importance of interactions between treatment variables and other factors.

Conclusions  Findings are discussed within the framework of a transactional model, and inferences are drawn about the limitations of the idea that there is a ‘best treatment’ that is universally applicable.