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-161- Wound Healing 9/5 Room-C
OCULAR SURFACE RECONSTRUCTION FOR SEVERE DRY EYE CORNEAL STEM CELL TRANSPLANTATION.
Kazuo Tsubota, Yoshiyuki Satake, Naoshi Shinozaki, Shigeto Shimmura, Jun Shimazaki.
Department of Ophthalmology, Tokyo Dental College, Chiba, Japan.
Ocular cicatricial pemphigoid (OCP) and Stevens-Johnson (SJS) syndrome often cause blindness and ocular damage which is not amenable to surgical correction. We present a surgical technique for reconstructing affected eyes. Fourteen eyes of 11 patients with cicatricial keratoconjunctivitis (7 OCP and 4 SJS) were treated with a combination of allograft limbal transplantation and frequent use of artificial tears derived from the patient's serum. With the mean follow-up time of 5 months, successful ocular surface reconstruction was achieved in 12 eyes with minimal recurrence of symblepharon. The concentration of EGF, vitamin A and TGF-¦Â in serum (essential tear components) were 0.51 +/- 0.08 ng/ml, 227.3 +/- 95.6 IU/dl, and 53.9 +/- 4.7 ng/ml respectively. These components were well preserved up to one month in the refrigerator (4 ¡î) and 3 months in freezer (-20 ¡î). The viability of corneal stem cells were confirmed in 3 cases by FISH method. We have shown that a combination of allograft limbal transplantation and the serum derived tears can reconstruct the ocular surface in most cases. Although the follow-up period was short and the number of the patients were small, this approach appears to offer an alternative to keratoprosthesis for treating severe cicatricial keratoconjunctivitis with dry eye.
-162- Wound Healing 9/5 Room-C
RECONSTRUCTION OF THE OCULAR SURFACE EPITHELIA OF PATIENTS WITH CICATRICIAL DISEASES BY AMNIOTIC MEMBRANE TRANSPLANTATION.
Jun Shimazaki, Hideo Sato, Naoshi Shinozaki, Kazuo Tsubota.
Department of Ophthalmology, Tokyo Dental College, Chiba, Japan.
Purpose: Amniotic membrane has been used as drapings in burns or after skin transplantation. Using its unique characteristics; anti-inflammatory effects and lack of immunogenecity, we propose a new surgical method for ocular surface reconstruction in patients with severe cicatricial diseases such as Stevens-Johnson syndrome . Methods: Amniotic membrane was obtained at the time of Cesarean sections and preserved at -80¡î. Immunohistochemistry for b-FGF, HGF, and TGF-¦Â, and quantitative analysis for the growth factors using ELISA were performed. Amniotic membrane transplantation (AMT) was performed for ocular surface diseases simultaneously as transplantation of stem cells of the corneal epithelium.Results:b-FGF, HGF, and TGF-¦Â were all present in the amniotic membrane.They were detected in the supernatant at concentrations of 5.31¡Þ3.04Ž¤2.45¡Þ1.45Ž¤14.24¡Þ5.96(ng/g), respectively. However, they all significantly decreased after two months' preservation. Clinically, AMT successfully reconstructed the ocular surface in about 70% of cases with severe cicatricial ocular diseases. Conclusions: Amniotic membrane seems to provide suitable microenvironment for normal differentiation and proliferation of ocular surface epithelia. AMT seems to be useful for the treatment of severe cicatricial ocular surface diseases.
-163- Wound Healing 9/5 Room-C
ANALYSIS OF HEALING PROCESS OF MRSA-INFECTED CUTANEOUS WOUNDS IN NORMAL AND DIABETIC MICE.
Ryoji Tsuboi, Chong-Ming Shi, Hideoki Ogawa.
Department of Dermatology, Juntendo University, Tokyo, Japan.
It is well known in clinics that bacterial infection delays the healing of chronic skin ulcers. In this study, we succeeded in the preparation of an infectious chronic wound model in diabetic mice by the application of MRSA (methicillin-resistant Staphylococcus aureus). Two three-mm diameter full thickness wounds were prepared on the dorsal skin of female C57BLKsJ db/db mice, 1I105 MRSA was applied locally, and the wounds then covered by a Cathereep film. Thirteen days after the application, the wounds were histologically evaluated by measuring reepithelialization (%), areas of granulation tissue (mm2), capillary numbers and the number of bacterial organisms. Results indicated that the diabetic mice had a high incidence of infectious ulcers and impaired healing when compared with those in normal mice, that the MSSA N315P and MRSA N315PZR strains had similar pathogenicity with respect to the formation of infectious wounds in diabetic mice, and that the treatment of infectious wounds with purified sucrose-povidone iodine is effective for reducing the number of the organisms and for promoting healing of the infectious wounds.
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THE EXPRESSION OF CONNECTIVE TISSUE GROWTH FACTOR DURING WOUND HEALING.
Nobukazu Hayashi1,2, Takashi Kakinuma2, Shigeru Kawara3, Kazuhiko Takehara3, Kunihiko Tamaki2, Atsuyuki Igarashi1.
1Division of Dermatology, Kanto Teishin Hospital, 2Department of Dermatology, University of Tokyo, Tokyo, 3Department of Dermatology, Kanazawa University Kanazawa, Japan.
Connective Tissue Growth Factor (CTGF) was known to be expressed in fibrotic diseases (ex. PSS and keloid) and tissue regeneration in wound model. But no report was existed which commented the CTGF expression on real wound sites. In this study We investigated CTGF expression on rat real wounds. We used 6mm Dispopunch to make ulcers on the back of rats, and tissue materials were taken on days 2, 4, 6, 8, 10, 12 after wounds.These tissues were used for in situ hybridization using Digoxgenin labeled CTGF probes. The result revealed that CTGF was continuously expressed on the fibroblasts around the wound. CTGF was more expressed on days 6 and 8. Also we found the CTGF message on the endothelial cells of the new vessels on wound site.