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34 STROMAL PROGENITOR CELLS SPC ; UPREGULATE SDF-1alpha SDF-1a ; PRODUCTION IN RESPONSE TO INFLAMMATION: A POTENTIAL MECHANISM FOR SPC MEDIATED ENHANCEMENT OF IMPAIRED WOUND HEALING A.T. Badillo, S. Chung, R.A. Redden, L. Zhang, E.J. Doolin, and K.W. Liechty, Children's Hospital of Philadelphia, Philadelphia, PA We have previously shown that SPC can improve wound healing in a murine model of diabetic wound healing. In addition, we observed decreased levels of SDF-1a in our untreated diabetic wounds and that treatment with SPC corrects this deficit. Chemotaxis of progenitor cells to tissues is regulated by interactions between SDF-1a and its receptor CXCR-4. Based on these observations we hypothesized that the correction of the diabetic wound healing defect by SPC treatment may, in part, be due to increased SDF-1a production and CXCR-4 receptor expression by SPC in response to signals in the early wound healing environment. To test this hypothesis we examined SDF-1a production and CXCR-4 expression by SPC in response to treatment with cytokines present in the early phases of cutaneous wound healing. METHODS: Fetal liver derived SPC were treated with increasing concentrations of PDGF, TGF-b1, TNF-a, or IL-10 over 48 hours. SPC production of SDF-1a was determined by ELISA and CXCR-4 receptor expression was determined by FACS analysis at both 24 and 48 hours. RESULTS: Stimulation with PDGF produced a 3 fold increase in SDF1a production over 48 hours. Stimulation with TGF- b-1caused a dose dependent increase in SDF-1a in the first 24 hours of treatment followed by down regulation to baseline expression levels after 48 hours of treatment. In contrast, treatment with the anti-inflammatory cytokine IL10 resulted in SDF-1a down-regulation. Stimulation with either PDGF, TGF b-1, or TNF-a increased CXCR-4 expression 2-4 fold. 35 A NOVEL MURINE MODEL FOR CREATING PARTIAL THICKNESS DERMAL INJURY TO STUDY PROGENITOR CELL ACTIVITY IN WOUND HEALING JA Greco III1, DL Ellis2, JM Davidson3, LB Nanney1Departments of Plastic Surgery1, Dermatology2, and Pathology3Vanderbilt University Medical Center--Nashville, TN Contributions of follicular bulge and circulating progenitor cells to wound healing have not been fully elucidated. Examination of these cells in mice requires creation of a superficial injury model that spares the bulge region. We hypothesized that the Free Electron Laser FEL ; would impart such a lesion with minimal collateral damage. Preliminary experiments on pelt and C57BL6 mice explored FEL parameters eg. wavelength, beam-path attenuation, and power ; needed to produce a consistent partial thickness lesion in mouse skin. Accordingly, at 12-14 mJ of energy the FEL created a reproducible partial thickness injury at a depth consistently superficial to the follicular bulge region in both murine strains tested. Depth of injury was quantified on days 0, 1, 2, and 6 days after wounding in C57BL6 control mice ; and MRL mice--a strain with a regenerative phenotype. Specimens were subjected to staining with H&E, Trichrome, and immunohistochemical stains Ki67, TUNEL, keratin 1, F4 80, CD 3, and CD34 ; to evaluate various cellular populations throughout the repair process. Ki67 immunostaining showed robust cellular proliferation emanating upward from the bulge region in both murine strains with differential effects on the rate of epidermal healing. MRL wounds showed an unexpectedly large infiltrate of mononuclear cells initially in the subcutaneous area. Proliferating cells in this population 12.6% ; reached a peak at 48 hours after wounding. A 4-fold increase in these was noted in the subcutaneous region over the healing dermal region at days 2 and 6 after wounding. Our data suggest that the FEL model is a reliable tool for creating wounds in mice that preserve and activate resident bulge stem cells and also incite an influx of circulating proliferating cells to the wound bed. Ongoing study is directed toward the use of additional markers to identify these mitotically active cells and their reparative roles. Work is supported with funds from the Departmentof Plastic Surgery. 36 HYPERBARIC OXYGEN PROTECTS AGAINST OXIDATIVE DAMAGE THROUGH DOWN-REGULATION OF MAPK SIGNAL TRANSDUCTION Q Zhang, Q Chang, W Myers, LJ Gould University of Texas Medical Branch, Galveston, TX Introduction: Hyperbaric oxygen HBO ; improves ischemic wound healing; the mechanism remains unknown. We have previously shown that HBO reduces apoptosis. In this study we examined the effect of HBO on the reactive oxygen species ROS ; mediated MAPK pathway in a rat ischemic wound model. Methods: Male Sprague-Dawley rats underwent creation of a validated ischemic wound. HBO treatment was compared to normoxia and to the free-radical scavenger, Nacetylcysteine. Four groups underwent daily treatment: HBO 90 minutes, 2.4 atm HBO NAC HBO plus 150mg kg NAC intraperitoneal NAC 150mg kg Control neither 6.
Pre-clinical studies focus on cytotoxicity in cell lines or explants What is an acceptable selectivity index for vaginally applied drugs? Rabbit vaginal irritation model Clinical trials rely on colposcopy & adverse events Experience with N-9 suggests this may not be sufficient Measurement of cytokines & other inflammatory markers may provide insight into potential toxicity of surfactant-type agents Precisely how or whether the inflammatory response to N-9 was responsible for the susceptibility to HIV unknown Biological functional significance of cytokine levels in vaginal secretions not defined, for example, zovirax uk.
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1. 2. 3. Hudson, T., Women's Encyclopedia of Natural Medicine. 1999, Lincolnwood, IL: Contemporary Publishing Group, Inc. Geller, S.E., Studee, L., Chandra, G., Knowledge, attitudes, and behaviors of healthcare providers for botanical and dietary supplement use for postmenopausal health. Menopause, 2005. 12 1 ; : 49-55. Ueda, M., A 12-week Structured Education and Exercise Program Improved Climacteric Symptoms in Middle-aged Women. Journal of Physiological Anthropology and Applied Human Science, 2004. 23 5 ; : 143-148. Lindh-Astrand, L., Nedstrand, E., Wyon, Y., Hammar, M., Vasomotor symptoms and quality of life in previously sedentary postmenopausal women randomized to physical activity or estrogen therapy. Maturitas, 2004. 48: p. 97-105. Dormine, S.L., Reame, N.K., Menopausal hot flash frequency changes in response to experimental manipulation of blood glucose. Nurse Res, 2003. 52 5 ; : 338-343. Christy, C.J., Vitamin E in menopause. J Obstet Gynecol, 1945. 50 84-87 ; . McLaren, H.C., Vitamin E in the menopause. Br Med J, 1949. ii: p. 1378-1381. Finkler, R.S., The effect of vitamin E in the menopause. J Clin Endocrinol Metab, 1949 9 ; : p. 8994. Barton, D.L., Loprinzi, C.L., Qella, S.K., et al., Prosepective evaluation of vitamin E for hot flashes in breast cancer survivors. J Clin Oncol, 1998. 16 2 ; : 495-500. Murase, Y., Iishima, H., Clinical studies of oral administration of gamma oryzanol on climacteric complaints and its syndrome. Obstet Gynecol Prac, 1963. 12: p. 147-149. Ishihara, M., Ito, Y., Nakakita, T., Maehama, T., Hieda, S., Yamamoto, K., Ueno, N., Clinical effect of hamma-oryzanol on climacteric disturbance on serum lipid peroxides. Nippon Sanka Fujinaka Gakkai Zasshi, 1982. 32 2 ; : 243-251. Yoshino, G., Kaxumi, T., Amano, M., et al., Effects of gamma-oryzanol on hyperlipidemic subjects. Current Ther Res, 1989. 45: p. 543-552. Pizzorno, J., Textbook of Natural Medicine 2 ed. ; . 1999, Edinburgh: Churchill Livingstone, Inc. Upton, R., Black Cohosh Rhizome, in American Herbal Pharmacopoeia and Therapeutic Compendium. 2002. Duker, E.M., Kopanski, L., Jarry, H., Wuttke, W., Effects of extracts from cimicifuga racemosa on gonadotrophin release in menopausal women and ovariectomized rats. Planta Med, 1991. 57: p. 420-424. Stolze, H., An alternative to treat menopausal complaints [in German]. Gyne, 1982. 1: p. 14-16. Warnecke, G., Influencing menopausal symptoms with a phytotherapeutic agent: successful therapy with cimicifuga monoextract [in German]. Med Welt, 1985. 36: p. 871-874. Stoll, W., Phytopharmacon influences atrophic vaginal epithelium: double-blind study: Cimicifuga vs. estrogenic substances [in German]. Therapeutikon, 1987. 1: p. 23-31. Mills, S., Bone, K., The Essential Guide to Herbal Safety. 2002, St. Louis, MO: Elsevier, Inc. Upton, R., Chase Tree Fruit, in American Herbal Pharmacopoeia and Therapeutic Compendium. 2001. Mills, S., Bone, K., Principles and Practice of Phytotherapy. 2000, Edinburgh: Churchill Livingstone. Hirata, J.D., Swiersz, L.M., Zell, B., Canchola, A.J., et al., Does dong quai have estrogenic effects in postmenopausal women? A double-blind, placebo-controlled trial. Fertil Steril, 1997. 68 6 ; : 981-986. Bone, K., Clinical Applications of Ayurvedic and Chinese Herbs. 1996, Blackburn, Victoria: Printgraphics Pty Ltd. Milligan, S.R., Kalita, J.C., Pocock, V., Van de Kauter, V., Stevens, J.F., Deinzer, M.L., Rong, H., De Keukeleire, D., The endocrine ativities of 8-prenylnaringenin and related hop Humulus lupulus L. ; flavonoids. J Clin Endorinol Metab, 2000. 85 12 ; : 4912-4915. Davydov, M., Krikorian, A.D., Eleutherococcus senticosus Rupr. & Maxim. ; Maxim. Araliaceae ; as an adaptogen: a closer look. J Ethnopharmacol, 2000. 72: p. 345-393. Tode, T., Kikuchi, Y., Hirata, J., Kita, T., Nakata, H., Nagata, I., Effect of Korean red ginseng on psychological functions in patients with severe climacteric syndromes. Int J Gynaecol Obstet, 1999. 67 3 ; : 169-174 and accupril.
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