bluuebunny/crossref_metadata_2025 huggingface dataset:
symposium II: basic research and clinical approach for impaired wound healing Moderated by Takashi Nakazuka: Negative‐Pressure Dressings in the Treatment of Infected Pressure Ulcers
Authors: Isago Tsukasa , Nozaki Motohiro
Published: January 2005

Applying negative pressure to wounds may speed the formation of granulation tissue, decrease the amount of localized edema, increase blood flow, and accelerate healing. In the present study, we treated 10 patients with stage IV chronic pressure ulcers associated with infection by this negative pressure dressings. The aim of this investigation is to study the efficacy of this therapeutic method for pressure ulcers with the infection by this negative pressure dressings. The long (A) and short (B) diameters of ulcer were measured to determine the size, and the vertical distance from the skin to the deepest point of ulcer was measured to determine the depth. Lesions were measured initially, at weekly intervals. The area of the lesion was taken to be 3.14 × A/2 × B/2 (cm2). In comparison with the area in the start and end of the treatment, the area had been reduced in all cases, and the reduction of 55.1% was shown from the area at the baseline. In comparison with the depth in the start and end of the treatment, the depth also decreased as well as the area in all cases, and the reduction of 61.2% was shown from the depth at the baseline. In view of the period of evaluation, the method is considered markedly effective in reducing the size and depth of ulcer.
Analysis of Pathophysiology of Pressure Ulcer in a Microcirculatory Model
Authors: Ichioka S , Tsuji S , Sekiya N , Nakatsuka T
Published: January 2005

Aim:  This study aims to establish a pressure ulcers model that visualizes the microcirculation, and to examine the participation of ischemia‐reperfusion injury in the pathophysiology of pressure ulcers.

Methods:  An original system composed of a new skinfold chamber and compression device allowed loading quantitative vertical stress to the skin. An intravital microscopic technique enabled direct visualization of the microcirculation in the physiological condition and in response to pressure application. To estimate the effect of ischemia‐reperfusion injury, animals were divided into two groups: the compression‐release group in which the animals received four cycles of compression‐release which consisted of 2 hours of compression followed by 1 hour of pressure release; and the compression alone group in which the animals underwent continuous compression for 8 hours. Functional capillary density was quantified before the compression procedure and on day1 (35 hours) after the first evaluation.

Results and Conclusions:  The cyclic compression‐release procedure significantly decreased functional capillary density as compared to continuous compression, indicating that in our experimental setting repetition of ischemia‐reperfusion cycle more severely damaged the microcirculation than single prolonged ischemic insult. The finding supports the significant contribution of ischemia‐reperfusion injury to the pathophysiology of pressure ulcers at the level of dynamic in vivo microcirculation.
How to Treat Acute Phase Pressure Ulcers with Necrotic Tissue
Authors: Tsukada Kunio , Tokunaga Keiko , Nagano Midori
Published: January 2005

Aim:  Most acute phase pressure ulcers have necrotic tissue. Choices to treat such ulcers are controversial. The aim of this study is to help produce a guideline to treat acute phase pressure ulcers with necrosis.

Methods:  Records of patients who acute phase ulcers with necrosis were retrospectively reviewed and their primary treatments and outcomes were studied. Ninety‐seven ulcers of 86 patients for 3.5 year period were included. Pictures of all ulcers were taken throughout the treatment periods.

Results:  Thirteen Stage II ulcers with black or yellow necrosis were treated with hydrocolloid dressings. All of them had excellent results. Forty of 76 Stage III ulcers had yellow necrosis. Hydrocolloid dressings were selected for 24 of them. Two of them became worth. Silver sulfadiazine cream was used for 5 ulcers and the results were relatively good. Fifteen of 36 ulcers with black necrosis were treated with hydrocolloid dressings. Two of them became worth. Thirteen ulcers with black necrosis were resected surgically at the beginnings of the treatments. One of them bled and became worth.

Only one of eight Stage IV ulcers had yellow necrosis and it was treated with hydrocolloid dressing and result was good. Remaining five ulcers with black necrosis were resected with good results.

Conclusions:  The primary choice for treating Stage II pressure ulcers with any type of necrosis is hydrocolloid dressings. We recommend hydrocolloid dressings for Stage III and IV ulcers without local symptoms of infection. Then after providing autolysis, surgical debridement should be done. If the patient is malnourished or is under poor pressure release, silver sulfadiazine cream may be recommended.
Oxygen Hyperbaric Pressure Therapy (OHPT) for Decubitus Ulcers
Authors: Fukai Takao , Nakata Masayuki , Takeda Akira
Published: January 2005

Aim:  Although oxygen hyperbaric pressure therapy (OHPT) for intractable ulcers is covered by national insurance, few studies have reported its use in the treatment of decubitus ulcers. In this study, we used OHPT as part of conservative treatment, and obtained good results.

Methods:  Five inpatients received a 60‐min session of 100% oxygen in a Hyperbaric Oxygen ChamberR(Kawasaki Engineering Co., Kobe, Japan) at 2 ATA for 10 days. The clinical response was evaluated by four parameters, the difference in the area of ulcer wounds before and after treatment, changes in inflammatory responses, state of wound infection, and histological findings.

Results:  OHPT was effective in reducing the area of ulcer wounds in four patients. Three patients, in whom blood tests had shown inflammatory responses (increases in WBC and CRP) before treatment, showed improvement. Two patients, in whom wound cultures were positive for MRSA, became negative after treatment. However, the patients varied in granulation tissue proliferation during treatment. The possible mechanisms underlying these changes were, first, improvement of tissue hypoxia by hyperbaric oxygen supply to ischemic tissue, leading to the above‐described reduction in wound area and improvement in inflammatory responses, and second, inhibition of bacterial growth due to an increase in active oxygen species, leading to the disappearance of wound infection.

Conclusions:  The results of this study suggest that OHPT is a very useful option in the conservative treatment of decubitus ulcers.
Chondrogenic and Osteogenic Differentiation of Adipose Tissue‐Derived Stem Cells Isolated from GFP Transgenic Mice
Authors: Ogawa Rei , Mizuno Hiroshi , Watanabe Atsushi , Migita Makoto , Shimada Takashi , Hyakusoku Hiko
Published: January 2005

Aim:  Taking advantage of homogeneously marked cells from green fluorescent protein (GFP) transgenic mice, we have previously demonstrated that bone marrow derived stromal cells (BSCs) differentiate into a variety of cell lineages both in vitro and in vivo. In the present study, we extend this approach to characterize adipose tissue derived stromal cells (ASCs), sometimes called processed lipoaspirate (PLA) cells.

Methods:  ASCs were isolated from inguinal fat pads of GFP mice. After the primary culture in control medium, the cells were incubated in osteogenic and chondrogenic medium for 2 to 4 weeks, respectively. Osteogenic and chondrogenic differentiation were assessed by the special staining and RT‐PCR.

Results:  ASCs incubated in osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. RT‐PCR analysis showed the Expression of osteocyte related genes. Positive staining cells for alcian blue and the expression of chodrocyte related genes were found in ASCs incubated in chondrogenic medium.

Conclusions:  These findings suggest that ASCs derived from GFP transgenic mice have both osteochondrogenic potential in vitro. Since this cell population can be easily identified through fluorescence microscope, it may be an ideal source of ASCs for further experiments of stem cell biology and tissue engineering.
Expression of the FGF Family and EGR‐1 in a Wound Healing Model (Reorganized Small Intestine From Fetal Mouse as an in Vitro Wound Healing Model)
Authors: Yonekawa Hironobu , Murata Eiko , Fujita Keiko , Satomi Akira , Akita Masumi
Published: January 2005

Aim:  In the present study, expression of members of the fibroblast growth factor (FGF) family and early growth response gene 1 (Egr‐1), known as a transcription factor, was analyzed by reverse transcription polymerase chain reaction (RT‐PCR).

Methods:  After the mouse fetal small intestine was divided into single cells with collagenase‐disperse, the cells were cultured. After reorganized small intestine were resected with a knife, expression of mRNAs from FGF family and Egr‐1 was analyzed by RT‐PCR.

Results:  The wound surface was covered with epithelial cells by 24 hours after resection. Among members of the FGF family, mRNAs from FGF‐1, ‐2, ‐5, and ‐7 were expressed. Egr‐1 was expressed as early as 15 minutes after resection. Egr‐1 is thought to be a transcription factor that induces the expression of FGF‐2, other than factors involving wound healing, such as PDGF and TNF‐a.

Conclusions:  In the present study, it was determined that this structure is a potential model for studying the roles of transcription factor and growth factors expressed in association with damage and wound healing.
Disturbance of Wound Healing Affects the Defensive Mechanism at the Base of the Gastric Mucosa
Authors: Yoshida Masashi , Wakabayashi Go , Ishikawa Hideki , Shimazu Motohide , Tanabe Minoru , Kumai Koichiro , Kubota Tetsuro , Otani Yoshihide , Saikawa Yoshiro , Kameyama Kaori , Kitajima Masaki
Published: January 2005

In vivo microscopy was performed to observe the basal region after thermal injury to the back skin of rats. A donor of nitric oxide, 3‐morpholinosydnonimine hydrochloride (SIN‐1), or a serine protease inhibitor, camostat mesilate, was administered. Antivascular endothelial growth factor (VEGF) neutralizing antibody was administered 5 hours after thermal injury (anti‐VEGF group). Postcapillary venules could be observed in the basal region of the gastric mucosa (PV‐BGM). The PV‐BGM was dilated 5 hours after thermal injury, and it was reduced by the administration of SIN‐1 or pretreatment with camostat mesilate. In the control group, the erosions did not reach the basal region of the gastric mucosa. Most of the erosions healed within 72 hours. Delayed healing was observed in the anti‐VEGF group. In the anti‐VEGF group, exudation and congestion in the basal region was observed at 24 hours, and ulcer formation was observed at 72 hours after thermal injury. The blood flow of the PV‐BGM is thus speculated to increase when the mucosal superficial circulation is disturbed. The PV‐BGM can contribute to the defensive mechanisms in the basal region of gastric mucosa. The abnormal process of the healing may disturb the defensive mechanism at the base of the gastric mucosa, thereby result in ulcer formation.
Mechanism of Impairment in Wound Healing of Electron Beam‐Irradiated Rat Skin
Authors: Sugiyama Kenji , Nakatsuji Eri , Ochiai Atsushi , Esumi Hiroyasu , Matsumura Yasuhiro
Published: January 2005

Aim:  Preoperative radiotherapy for head and neck or colon cancer frequently prevents the following surgical operation, since effective dose of ionized radiation induces wound complication. To make the operation possible, causes for the impaired wound healing were clarified.

Methods:  Thighs of F344 rats were irradiated with electron beam (30 Gy). Seven days after the irradiation, full‐thickness skin of the thigh was cut by punch biopsy (4‐mm diameter). Healing process was determined by measuring size of wound opening. Cytokeratin 14, smooth muscle actin (a marker of myofibroblasts), and integrin beta1 were detected by immunohistochemical analyses.

Results:  The wounds in the nonirradiated rats healed almost completely on day 7, whereas repair of epidermis was impaired in the irradiated skin. Numbers of hair follicles and micro‐vessels decreased, and myofibroblasts were almost missing in the scar and dermis adjacent to the wounds of irradiated skins. Proliferation indexes of epidermal basal cells were much higher in irradiated skins than those in nonirradiated skins, however, these basal cells did not express cytokeratin 14, concomitant with the absence of integrin beta1 in basement membrane of the irradiated epidermis. On the other hand, apoptosis appeared in the dermal follicular cells of hair bulbs, 18 h after the irradiation. On day 3, structures of hair follicles were destructed.

Conclusions:  Reepithelialization of wounds was blocked in the irradiated skin. Since epidermal basal cells are supplied from stem cells in the hair bulge during cutaneous wound healing, one of causes for the impaired wound healing may be insufficiency of epidermal stem cells.
Preventive Effect of Fibroblast Growth Factor and Hepatocyte Growth Factor on Ceramide‐Induced Dysfunction Human Small Intestinal Cells
Authors: Yamaguchi Ryo , Takami Yoshihiro , Iida Takuya , Shimazaki Syuij
Published: January 2005

Purpose:  Gastrointestinal (GI) damages associated with multiple organ failure is a fatal complication. The injured function of GI tract causes malnutrition and the bacterial translocation. So the functional recovery of GI tract is quite important for treating such cases. In the present study, we investigated whether basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) could recover the injured functions of cultured intestinal cells.

Method:  Human small intestinal cells were cultured in a 10% FCS added DMEM. The cellular damages were induced by the treatment with various concentrations of cell permeable C6‐ceramide (0–20 M). Growth factors (10 ng/ml bFGF or 50 ng/ml HGF) were added to the medium simultaneously or 2 days after the beginning of the ceramide treatment. After 4 days of the treatment, cell growth and apoptosis induction were investigated.

Result and Conclusion:  Ceramide inhibited cell growth and induced apoptosis in a dose‐dependent manner. Both growth factors significantly improved the cell growth and reduced apoptosis. These effects were also observed when the growth factor treatment was delayed 2 days after the ceramide treatment. It was found that bFGF was more significant than HGF on these protective effects. It is suggested that growth factor treatment may be a useful therapeutic option to improve GI tract dysfunction.
General session III Moderated by Takehisa Iwai: Muscular Flap Treatment for Intractable Ulcer with Artificial Vessel Exposure After a Surgery for Thoracic Aortic Aneurysm
Authors: Yamauchi T , Kiyokawa K , Tai Y , Inoue Y , Kizuka Y.
Published: January 2005

Aim:  To heal an exposed artificial vessel due to mediastinitis after the replacement of thoracic aortic aneurysm.

Methods:  The exposed vessel is washed thoroughly, wrapped in a muscular flap, and a free skin graft is grafted over it to close.

Results:  An 81‐year‐old male repeatedly had mediastinum after the replacement of aneurysm, and it was not cured with muscular fillings, and a rubor on the chest developed to a subcutaneous hematoma. The lesion was closed with a patch on the artificial vessel, the mediastinal abscess was washed, and the opening was closed by using latissimus dorsi muscular flap and split‐thickness skin graft. No recurrence has occurred for 3 years. A 69‐year‐old male developed mediastinitis after the replacement of thoracic arch aortic aneurysm, and the wound reopened and the vessel exposed. Because conservative treatment did not help, pectoralis major muscular flaps with the perforating branches of the internal thoracic artery and vein as the pedicle were elevated from the both sides, filled the mediastinal space and a split‐thickness skin graft was grafted. No recurrence has occurred for one year.

Conclusions:  Artificial vessels that developed infections are usually removed. However, we had good results by washing the lesion and wrapping it with muscular flap. By considering the size of surgical invasion and high death rate associated with the re‐replacement of artificial vessel or the opening of omentum, utilization of muscular flap should be considered as the first choice treatment.
Wound Healing and Scar Formation on the Skin of the Chest Wall and the Lower Leg After Cardiac Bypass Surgery
Authors: Takahashi Hajime , Kurata Daisuke , Takeuchi Masaki , Sasaki Kenji , Nozaki Motohiro
Published: January 2005

Introduction:  Many patients receiving heart surgery have extensive operation scars on the skin surface of the chest wall and lower leg. In some cases there are long lasting open wounds in the lower leg after vein harvesting. Some patients complain of pain and itching and we can recognize hypertrophic or extensive scars, which we have to treat.

Case and result:  We treated 16 patients (11 men and 5 women) ranging in age from 47 to 86. They were receiving many drugs such as anticoagulants, thus wound healing was delayed. In these cases, we choose not to resect tissue but apply ointment to remove necrotic tissue, and prepare good granulation tissue for smooth epithelization. Then, we employ prophylactic measures against hypertrophic scar formation such as tape containing steroid, silicon gel sheets and skin tapes as well oral medicine (toranilast) for the same purpose. Because of they have had to continuously take many oral medicines, these patients often do not want to take additional oral medicine. This is one point of difference in the treatment of heart surgery patients and patients with other types of wounds.
Involvement of KGF in the Wound Healing Process of Tracheal Cartilage in Rat Longitudinal Tracheotomy Model
Authors: Abo Takafumi , Matumoto Keitarou , Yamayosi Takatomo , Nagayasu Takesi , Izumi Sinich , Hisikawa Yositaka , Kouji Takehiko
Published: January 2005

Aim:  To address whether KGF (Keratinocyte Growth Factor) and KGFR (keratinocyte Growth Factor Receptor) expression is involved in the wound healing process of tracheal cartilage in rat longitudinal tracheotomy model. And to expose the new strategy to control the process utilizing KGF and its related compounds.

Method:  In this study, we incised rat trachea longitudinally for three cartilage lings, and harvested trachea from rat on 0, 1, 3, 5, 7 days after operation. We made horizontal section and observe them with immunohistochemistry.

Result:  The cartilage defect was filled with chondrocytes originated from the perichondrium. KGF expression was observed in chondrocytes, perichondrium and fibroblasts in injured area. The expression of KGF was upregulated immediately after the operation, reaching a peak at 3–5 days.

Future approach:  Further examination on temporal and spatial pattern of KGF and KGFR expression. Quantitative analysis of the expression of KGF and KGFR in relation to cell kinetics. Attempts to manipulate the kinetics of wound healing using KGF and its related compounds.
General session IV Moderated by Yoka Yoshimura: A Retrospective Study of Vacuum‐Assisted Closure in Japan
Authors: Miyamura Suguru , Terashi Hiroto , Yanagi Hideyuki , Tsuji Yoriko , Nomura Tadashi , Nonomura Hideaki , Tahara Shinya
Published: January 2005

Aim:  Vacuum‐assisted closure (VAC) system, a new method for wound treatment, has been widespread over the world. There are few reports about this treatment in Japan. The purpose of this report is to consider the usefulness of VAC treatment at our department.

Methods:  A retrospective review of 24 patients with 27 chronic wounds, which have been treated with VAC, was presented. The following parameters were studied: age, gender, present illness, complications, duration of VAC treatment, and its result.

Results:  Eight women and 16 men were presented. The median age was 66.1 years (range, 20–84 years). Eleven of 27 wounds were mediastinitis. Six were pressure ulcers, and nine were other complicated ulcers. Tweleve patients had diabetes mellitus, and nine had hypertension. The median duration of VAC treatment was 32.5 days. Tweleve wounds healed only with VAC, and nine healed with a combination of VAC and small operation. Six were halfway stopped a VAC treatment due to infection.

Conclusions:  Our results suggest that VAC system is an alternative treatment for chronic wounds of elder patients with high risk.
Chemical Analysis of the Surface of Silicone Modified by Oxygen Plasma Treatment for Cell Adhesion
Authors: Yamada Tai , Ohsugi Ikuko , Inoue Yoshikazu , Mizuno Kiyoyuki , Okumoto Takayuki , Yoshimura Yohko
Published: January 2005

Aim:  As we confirmed O2 plasma treatment is useful for cell adhesion and growth on silicone surface by the culture experiment, reported in the other paper (Ohsugi et al). We aimed to confirm the change of the surface of silicone by O2 plasma treatment from the material‐scientific view.

Material:  All materials were cut from a medical grade silicone mammary implant as the same sized disks. Group 1 is silicone disks with no treatment on material's surface. Group 2 is silicone disks with surface modifications by O2 plasma treatment for 3 minutes. Group 3 is silicone disks with surface modifications by O2 plasma treatment for 10 minutes.

Method:  We investigated each group by the scanning electron microscope (SEM), the atomic force microscope (AFM), and X‐ray photoelectron spectoroscopy (XPS).

Result:  As SEM and AFM shows, after O2 plasma treatment for 3 minutes, an original inequality formation (10 μm in width) on a surface of silicone were made flat, and for 10 minutes a crack formation occurred.

From investigation by XPS, after O2 plasma treatment for 3 minutes, the ratio of oxygen on the surface of silicone increased. This phenomenon was thought that hydrophilic functional groups (‐OH, ‐COOH, etc.) were grafted on the surface of silicone. We supposed this is the reason why cell adhesion and growth became more effective.
Effect of Oxygen Plasma Treatment for Cell Adhesion Properties on Silicone Surface
Authors: Ohsugi Ikuko , Yamada Tai , Inoue Yoshikazu , Mizuno Kiyoyuki , Okumoto Takayuki , Yoshimura Yohko
Published: January 2005

Aim:  The silicone polymer is known to be useful and safety as a material of medical implants. But fundamentally its biocompatibility and biostability are questionable. Thus we try to make a hybrid‐type silicone implant coating by cell layers. At first, we aimed to confirm cell adhesion and growth on silicone surface.

Material:  Group 1 is silicone implants for the augmentation rhinoplasty with no treatment. Group 2 is same type silicone implants with surface modifications by O2 plasma treatment (3 min). Group 3 is the group 2 implants with serum and plasma coating (20 min).

Method:  We co‐cultured materials of each group with human fibroblast (2.4*104 cells/ml), for 17 hours, with 5%CO2, in the temperatures of 37 °C.

Results:  Group 1; none of fibroblasts could take the surface of materials.

Group 2; several fibroblasts were found on the surface of those.

Group 3; much more fibroblasts were found than group 2

This result means that the surface modification by O2 plasma treatment itself enables the cell adhesion and growth on silicone surface.
Comparison of Bacteria‐Retaining Ability of Wound Dressings
Authors: Tachi Masahiro , Hirabayashi Shinichi , Yonehara Yoshiyuki , Uchida Gentaro , Tohyama Takuya
Published: January 2005

We studied the bacterial retention capacity of alginate and carboxymethylcellulose dressings, using an infected skin ulcer model on the backs of rats. Wound surfaces were inoculated with either Staphylococcus aureus or Pseudomonas aeruginosa at a concentration of 1.5 × 106 cfu/wound. Carboxymethylcellulose dressing, alginate dressing A and alginate dressing B were applied to the contaminated wounds for 12 hours. Each dressing was then divided into two piece. Total viable bacterial count within the dressing was calculated using one piece, and bacterial count released from the dressing into physiological saline was determined using the other piece, enabling bacterial retention rate to be calculated. Each dressing was tested on each of 10 wounds. Statistical analyses were performed using one‐way analysis of variance (ANOVA) for replicated measures combined with Duncan's Multiple Comparison Test. Carboxymethylcellulose dressing was most effective in its ability to retain both Staphylococcus aureus and Pseudomonas aeruginosa(p < 0.05).

Conclusions:  The bacterial retaining ability of carboxymethylcellulose dressing was found to be significantly higher than that of alginate dressings in an infected animal wound model.
Chitosan Sponge with Photocrosslinkable Chitosan Hydrogel Stimulates Large and Impaired Wound Healing in Rats
Authors: Ishizuka Toshiaki , Ishihara Masayuki , Obara Kiyohaya
Published: January 2005

Aim:  We have previously reported that a photocrosslinkable chitosan hydrogel (Az‐CH‐LA) has induced wound healing in diabetic mice. However, deep and large skin wounds may be not enough covered by Az‐CH‐LA only. Therefore, we evaluated whether chitosan sponge with Az‐CH‐LA accelerates large and impaired wound healing.

Methods:  Full thickness‐square wounds (25 × 25 mm) were prepared on the upper back of twenty‐four male Sprague Dawley rats. Chitosan sponge and/or Az‐CH‐LA aqueous solution (30 mg/ml) was added onto the wound of each rat and was then irradiated with UV light. Thus, the following four experimental groups were studied: Group (G) 1: control, G2: Az‐CH‐LA solution only, G3: Chitosan sponge + 0.9% saline, or G4: Chitosan sponge + Az‐CH‐LA solution. The changes in wound area were measured at day 2, 4, 8, and 16. The skin including the wound was removed from each rat for histological examination.

Results:  The wound area in G4 at day 2, 4, and 8 was significantly smaller than the other groups. The number of neovessels and the thickness of granulation tissue in G4 at day 2 and 4 were significantly higher than the other groups.

Conclusions:  The chitosan sponge with Az‐CH‐LA has been found to be more effective in large and impaired wound healing compared with a Az‐CH‐LA solution only.
Liver Regeneration After Resection of Multiple Liver Cysts
Authors: Suehiro Taketoshi , Hirashita Teijirou , Mashino Koujirou , Sunmiyoshi Yasushi , Takeuchi Hideya , Aramaki Masanori , Watanabe Akihiro , Kanoh Tadashi
Published: January 2005

Background:  A regenerative function is in one of the functions of liver, and if it is a normal liver, it will reproduce even to capacity almost before an operation within one year after excision. However, liver substance is pressed over the long period of time, and, as for liver cysts, a postoperative liver reproduction process is unknown. Comparison examination of the multiple liver cysts postoperative liver reproduction was carried out with after the operation.

Patients:  It was aimed at one example of a multiple liver cysts operation and three metastatic live tumor (two extended left lobectomy, one right lobectomy) in this hospital. Liver volume was measured before an operation and 14 postoperative days CT, and comparison examination of the standard liver volume ratio called for from height weight was carried out.

Results:  Before an operation of liver cysts and 14 postoperative day, liver volume was 151% 45 or 69% in 450 or 680 ml. Before an operation of control group and day 14 postoperative day liver volume was an average of 128% at 116 and 128 or 140% in 62, 72, 56, 71, and 49 or 68% at 618, 720, 532, 680, and 516 or 720 ml, respectively.

Conclusion:  Multiple liver cysts postoperative liver reproduction was promoted compared with after the liver resection.
General session VI Moderated by Nobutoshi Andou: Lung Vitamin E Depletion Resulting from Burn and Smoke Inhalation in Sheep
Authors: Shimoda Katsumi , Soejima Kazutaka , Isago Tsukasa , Nozaki Motohiro , Traber Daniel L.
Published: January 2005

Introduction:  Oxidants are involved in the pathogenesis of multiple trauma caused by burn and smoke inhalation. α‐ and γ‐tocopherol are major tissue antioxidants and their depletion reflects oxidant injury. We hypothesized that lung vitamin E levels in sheep would be depleted with multiple trauma.

Methods:  Sheep (n = 31) were surgically prepared for chronic study, then received either a 40% body surface area third‐degree burn, 48 breaths of cotton smoke, a combination of these injuries (B&S) or were given sham injuries. All sheep were resuscitated with Ringer's lactate solution (4 ml/kg/%BSA burn) and mechanically ventilated. The sheep were sacrificed at various time intervals as indicated and vitamin E was measured (nmol/g) in snap‐frozen lung samples obtained at autopsy 48 h postinjury.

Results:  Lungα‐tocopherol was significantly depressed in sheep with the combination injury, smoke alone and burn alone, respectively, compared to sham treated (p < 0.05). Lungγ‐tocopherol was significantly depressed in sheep with the combination injury compared to sham treated (p < 0.05). Neither smoke alone nor burn alone animals were statistically different from sham.

Conclusion:  α‐tocopherol depletion may be a result of lipid peroxidation, while γ‐tocopherol depletion may reflect both lipid peroxidation and peroxynitrite damage (nitro‐γ‐tocopherol formation). These results suggest that animals receiving a combination burn and inhalation injury have undergone marked oxidative stress.
General session VII Moderated by Chifumi Yoshida: A Case of Ulcerative Colitis with Peristomal Pyoderma Gangrenosum
Authors: Ebata Tomoe
Published: January 2005

Introduction:  Pyoderma gangrenosum is a skin disease that can result in progressive necrosis and formation of painful ulcers. This is to report on a case where the patient developed the disease around a stoma.

Case:  A 40‐year‐old female patient developed ulcerative colitis (UC) when she was 25. At the age of 38, the exacerbation of UC required a colectomy and ileal pouch anal canal anastomosis (IACA), following which, a permanent ileostomy was constructed due to pouchitis. In April 2003, she developed redness around the stoma with pain, which resulted in the gradual formation of an ulcer. She was hospitalized with a diagnosis of peristomal pyoderma gangrenosum (PPG). After admission to the hospital, the dosage of predonine was increased to 30 mg/day, and dapsone was concomitantly administered. Clobetasol propionate and sulfadiazine silver were topically administered 1–4 times daily. The wound region adjacent to the stoma was protected with a gauze dressing. Although the gauze was changed each time, it was contaminated with fecal matter, the suffering thereby inflicted on the patient was too severe due to the associated smell and staining of her clothing. For this reason, she alternatively started wearing a stoma bag, which enabled periodic replacement once daily. Following a gradual improvement of the ulcer, she was discharged from the hospital two months after admission. Her recovery was confirmed four months after onset.

Conclusion:  PPG is accompanied by pain, and treatment also involves suffering by the patient. We need to make the diagnosis and determine the treatment method while the disease is still in its early stages. The selection of the method of care should be made with consideration of the practical aspect of the patient's defecation control.
Important Factors in the Healing of a Pressure Ulcer in the Ischium of a Spinal Cord‐Injury Patient Whose Pressure Ulcers Recurred Over 26 Years
Authors: Sugai Ayumi , Yamamoto Hiromasa
Published: January 2005

Objectives:  Pressure ulcers in the ischium of spinal cord‐injury patients are caused by their wheelchair‐bound lifestyle. Such injuries are intractable and limit a patient's activities of daily life for long periods. Here, we report our experience with a patient whose pressure ulcer was healed without limitation of her daily activities.

Subject:  A woman aged 56, who had injured her thoracic spinal cord 26 years earlier, developed a pressure ulcer in the right ischium. The injury healed and then recurred over 10 admissions for treatment, and the maximum period over which the patient stayed at home without a pressure ulcer was only two months over the past several decades. In June 2002, she was healed and discharged, but the pressure ulcer recurred in the same region only one month later.

Course:  She was treated in the outpatient department, but the affected area expanded and the condition aggravated and invaded the subcutaneous tissue. The causes were considered to be a body posture leaning to the right, rubbing during decompression, and pressure on the lesion exerted by a cushion of the wheelchair. Treatment of the patient began from a consideration of daily activities, including work and housekeeping. The cushion was changed to a new one that helped the patient maintain a good posture and prevented rubbing. In addition, the patient was instructed to develop a decompressing action that dispersed the body pressure to the posterior thigh, and was shown how to sit during monthly visits to the outpatient department. The affected lesion was washed every day in the shower, and was treated with sugar and povidone‐iodine.

Results:  The patient observed changes in the affected lesion of the pressure ulcer, and reported changes that occurred in her daily life. In April 2003, the pressure ulcer had healed and has yet to recur 8 months later.

Conclusions:  (1) For pressure ulcers in the ischium of a spinal cord‐injury patient, it is of importance for healing and prevention of recurrence to find the cause of the pressure and to improve the condition, rather than to treat the affected lesion. (2) Instructions regarding adoption of the correct sitting position, i.e., a major factor in daily life, and continuous care intervention improved the patient's self‐care ability.
symposium i‐1. pathophysiology and treatment of pressure ulcer Moderated by Koicki Hirata: The Study of Clinical Usefulness of Basic FGF
Authors: Hamanoue Masahiro , Nakajo Tetuhiro , Kijima Fumio , Nisijima Hiro , Maenohara Sigeho
Published: January 2005

Recently, basic FGF (fibroblast growth factor) has been made clinical use for wound cure. We studied clinical usefulness of basic FGF.

Cases:  Nine cases took wound necrosis by infection. Two cases took burn after mastectomy. One case took abscess in pelvic space.

Methods:  We investigated healing rate of wound by healing area per day. And we evaluated histological wound healing by HE staining, labeling index of Ki64 staining, CD34 staining for index of neovascularization, and α‐smooth muscle actin staining for differentiation of fibroblasts.

Result:  Healing rate of nine cases with wound necrosis by infection was 5 cm2/day and that of two cases with burn after mastectomy was 0.5 cm2/day. Labeling index was more than 10%. CD34 staining showed increasing neovascularization of healing tissues with treatment of bFGF, compaired with that recieved other treatments. But the case with abscess did not recover and took the reoperation for treatment.

Conclusion:  We concluded that bFGF was efficacious against tissue damage and useful for wound cure. But basic FGF was not useful for the infectious focus without control.
European Tissue Repair Society Focus Meeting Tissue Repair, Contraction, and the Myofibroblast Nyon, Switzerland November 18–20, 2004 The following compilation of abstracts represents a partial list of submissions received for presentation at the meeting. Please note the abstracts are arranged in alphabetical order (by presenter's name). Communications by the invited speakers will be published in a book by Landes Bioscience.: TGF‐β1 Signaling is Regulated by Negative Feedback Mechanism of BMP‐7 in Fibrotic Kidneys of Hereditary Nephrotic Mice
Authors: Anan S , Goto Y , Yamaguchi‐Yamada M , Sakata C , Manabe N
Published: January 2005

ICR‐derived strain with gulomerulonephrisis (ICGN) mouse derived from ICR mouse is a newly established mouse strain with hereditary nephrotic syndrome. The affected ICGN mice consistently show the clinical signs of nephrotic syndrome, developing various degrees. The lesions consist of thickened capillary basement membrane, enlarged mesangium without cellular proliferation, and tubulointerstitial fibrosis. Based on these histopathological findings and clinical biochemical data, ICGN strain mouse is considered to be a good experimental model for human idiopathic nephrotic syndrome. Extracellular matrix (ECM) components provide structural integrity to the tissues, and mediate regulation of cell growth, migration, differentiation, etc. The damaged cells have increased secretion of ECM components. This is a kind of self‐protecting reaction that is associated with healing process against injury. However, the hyperaccumulation of ECM components leads to fibrosis, a pathological phenomenon. Therefore, it is very important to make clearly the process of fibrosis in ICGN strain mouse. In this study, the expression and localization of transforming growth factor beta1 (TGF‐β1), TGF‐β1 type I receptor (TGFR1), and bone morphogenic protein (BMP)‐7, which regulate the signals of TGF‐β1 negatively, were investigated by using immunohistochemical staining and Western blotting. High expression levels of TGF‐β1 and TGFR1 were observed in tubular epithelial cells of middle stage‐ICGN mice as compared with those of control ICR mice. BMP‐7 was strongly expressed in tubular epithelium of ICR mice, but it decreased with the progression of renal disease in ICGN mice. Moreover, no localization of BMP‐7 in expanded tubular epithelial cells in ICGN mice was seen. These data suggest that the increase of TGF‐β1 and TGFR1 and the degradation of negative regulation of TGF‐β1 by BMP‐7 in middle and late stages of ICGN mice play an important role in renal fibrosis of ICGN mice. The present findings may contribute to elucidate the pathogenic mechanisms of human idiopathic nephrotic syndrome.
The Role of Matrix Stiffness in Hepatic Stellate Cell Activation and Liver Fibrosis
Authors: Gaca MDA , Georges PC , Hui JJ , Janmey PA , Wells RG
Published: January 2005

Transdifferentiation of hepatic stellate cells into proliferative and fibrogenic myofibroblasts is a critical event in the development of liver fibrosis. The causes of HSC transdifferentiation are not well understood, although the extracellular matrix has been implicated as one possible factor. We hypothesized that HSC transdifferentiation is determined by the mechanical properties of the surrounding matrix, and that the diseased liver demonstrates an increased elastic modulus that contributes to the perpetuation of HSC transdifferentiation and the progression of fibrosis. To test this hypothesis in an in vitro system, primary rat HSC were plated on inert polyacrylamide supports of variable but precisely defined elastic modulus ranging from 100 to 12 K Pa. These were coated with different matrices (type I collagen, fibronectin, or Matrigel™) in layers thin enough not to alter the mechanical properties of the system. As determined by morphology, degree of cell spreading, and expression of α‐smooth muscle actin, cells became progressively transdifferentiated as substrate stiffness (elastic modulus) increased, independent of the chemical identity of the coating matrix. The degree rather than speed of HSC transdifferentiation correlated with substrate stiffness, with cells cultured on supports of intermediate stiffness adopting stable intermediate phenotypes. These changes were independent of the addition or removal of TGF‐β. This demonstrated in an in vitro system that HSC transdifferentiation is primarily a function of the physical rather than the chemical properties of the substrate. As a preliminary to determining the role of stiffness in vivo, we determined the elastic modulus of normal and progressively fibrotic rat liver. Rats were injected twice weekly with carbon tetrachloride or vehicle, and were sacrificed in groups of 7–8 (2 controls and 5–6 CCl4) at time periods ranging from four days to ten weeks after the first injection. Control rats demonstrated a G’(elastic modulus) of 460 +/− 25 Pa. The mean elastic modulus of CCl4‐treated livers increased progressively, beginning at the four day point, 525 +/− 39 Pa, and reaching 1617 +/− 170 Pa at 10 weeks. These values are consistent with the values at which we saw intermediate degrees of HSC transdifferentiation in vitro, and suggest that alterations in liver stiffness are a key factor driving the progression of fibrosis.
Mechanical Stress Induces TGFβ1 Activation in Myofibroblast Culture
Authors: Wipff PJ , Braunecker J , Meister JJ , Hinz B
Published: January 2005

Differentiation of fibroblasts into contractile, α‐smooth muscle actin (α‐SMA) expressing myofibroblasts depends on the action of the cytokine TGFβ in conjunction with mechanical tension. The goal of this study was to assess whether mechanical stress may play a role in activating TGFβ. Previous studies have shown that myofibroblasts secrete TGFβ1 as a large latent complex, consisting of the latency associated protein (LAP), the latent TGFβ1 binding protein (LTBP‐1) and TGFβ1. LTBP‐1 targets the small latent complex of LAP and TGFβ1 to the extracellular matrix (ECM), providing a stock of latent TGFβ1. To reveal the mechanisms triggering the release of active TGFβ1 from LAP and LTBP‐1, myofibroblasts were cultured on flexible silicone membranes for 3d before being subjected to unique 5% uniaxial stretch. After stress application we observed the course of TGFβ1 activation over 24 h by quantifying luciferase synthesis under the control of the TGFβ‐inducible PAI‐1 promoter. TGFβ activation showed two distinct peaks 1 h and 6 h after stretch. TGFβ1 mRNA levels were increased after 6 h as assessed by semi‐quantitative RT‐PCR. To further determine if rapidly activated TGFβ1 was released from ECM stores, we stretched myofibroblast‐derived ECM after cells have been removed by desoxycholate or EDTA treatment. Finally, we assessed the importance of the actin cytoskeleton in TGFβ1 activation by stretching Triton‐X‐100‐ and cytochalasin‐D‐treated myofibroblasts. Our results suggest that mechanical activation of TGFβ in myofibroblast culture requires cell activity and a functional cytoskeleton.
Increased Pyridinoline Cross‐Link Levels in Fibrosis is Accomplished by TGFβ Induced Lysyl Hydroxylases 2B Expression
Authors: Van Der Slot AJ , Zuurmond AM , Bank RA
Published: January 2005

Fibrosis is a complex process involving an acute inflammatory response, which finally results in the excessive deposition of collagen. The deposited collagen shows an increase in pyridinoline cross‐links, due to overhydroxylation of lysine residues within the telopeptides. As we have found that the enzyme responsible for the hydroxylation of the telopeptide lysine residues is lysyl hydroxylase 2 (LH2)[1], it was examined whether LH2 expression is increased in fibrotic lesions. Furthermore, we examined whether the expression of LH2 can be induced by TGFβ, a cytokine playing an important role in fibrotic processes. LH2a and LH2b (two splice variants of LH2) mRNA expression was analyzed in fibroblasts isolated from keloid, hypertrophic scars, fibrotic lesions of Dupuytren's and systemic sclerosis patients, as well as in activated stellate cells, which are responsible for liver fibrosis. Furthermore, LH2b expression was examined in control fibroblasts stimulated with TGFβ1, TGFβ2 and TGFβ3. The matrices deposited by the fibrotic fibroblasts as well as by the TGFβ stimulated control fibroblasts were analyzed for the presence of pyridinoline cross‐links. All fibrotic fibroblasts evaluated showed increased LH2b mRNA expression, which was associated with elevated pyridinoline cross‐link levels in the extracellular matrices deposited by these fibroblasts. LH2a was hardly detectable in all cells, suggesting that this splice variant does not play a significant role in fibrotic conditions. These data indicate that in fibrotic lesions, LH2b is responsible for the overhydroxylation of the collagen telopeptides. TGFβ1, TGFβ2 and TGFβ3 were able to induce LH2b expression. In addition, the three TGFβ isoforms increased the pyridinoline cross‐link levels in the matrices deposited by the stimulated cells, showing that TGFβ promotes the formation of pyridinoline cross‐links via the induction of LH2b expression. We conclude that LH2b plays a key role in fibrotic processes via the formation of pyridinoline cross‐links, giving rise to collagen that is difficult to degrade and therefore significantly contributes to the collagen accumulation.
Microscopical Characterization of Poly‐L/D‐Lactide (PLDLA) 96/4 Sutures in the Achilles Tendon of Rabbits
Authors: Kangas J , Pajala A , Leppilahti J , Ryhänen J , Länsman S , Törmälä P , Waris T , Ashammakhi N
Published: January 2005

Common Achilles tendon ruptures are usually not fixed by absorbable sutures due to limitations in their strength properties. Modern technology has made it possible to develop bioabsorbable sutures with prolonged strength retention. To evaluate histologically tissue reactions and biodegradation of poly‐L/D‐lactide (PLDLA) sutures implanted in Achilles tendon of rabbits. Fifteen rabbits were operated on and killed within a time schedule of 2, 6 and 12 weeks, with five rabbits in per period. PLDLA monofilament sutures (Tampere University of Technology, Tampere, Finland) were implanted inside the rabbit medial gastrocnemius tendon for biocompatibility testing. Polyglyconate (4.0) monofilament sutures (Maxon®, Cyanamid of Great Britain Ltd., Gosport, UK) with the same diameter were implanted in the contralateral tendon. The histology was studied in hard‐resin embedded samples and the thickness of the encapsule membrane was determined histomorphometrically.

PLDLA have significantly smaller capsule formation around each fiber in tendon than Maxon® sutures. The mean capsule thicknesses around PLDLA sutures was 5.26, 11.66, and 10.63 after 2, 6, and 12 weeks respectively. The mean capsule thicknesses around Maxon® sutures was 13.22, 80.97, and 17.59 after 2, 6, and 12 weeks, respectively. In conclusion, by 12 weeks the PLDLA sutures implanted intratendineously formed thinner fibrous capsule than Maxon® sutures of same diameter. The suture materials were not degraded by 12 weeks.

Acknowledgments:  Research funds from the Technology Development Center in Finland (TEKES, Biowaffle Project 40274/03 and MFM Project 424/31/04), the European Commission (EU Spare Parts Project QLK6‐CT‐2000‐00487), the Academy of Finland (Project 73948) and the Ministry of Education (Graduate School of Biomaterials and Tissue Engineering) are greatly appreciated.
Smooth Muscle Cells of Fatal Coronary Artery Lesions Exhibit a Myofibroblastic Phenotype
Authors: Hao H , Gabbiani G , Camenzind E , Virmani R , Bochaton‐Piallat ML
Published: January 2005

Objectives:  Characterize the phenotypic features of smooth muscle cells (SMCs) of erosions, stable plaques and in stent restenosis causing sudden death, in order to verify the possibility that they have modulated into myofibroblasts.

Methods and Results:  Expression of α‐SM actin (α‐SMA), SM myosin heavy chains (SMMHCs) and smoothelin was investigated by immunohistochemistry followed by morphometrical quantification in the media and intima of coronary lesions. The expression of all cytoskeletal proteins was lower in the media of restenotic lesions and, to a lesser extent, of stable plaques compared to that of erosions demonstrating a significant medial atrophy in the two first conditions. An important expression of α‐SMA was detected in the intima of the different lesions; moreover, the area of α‐SMA staining was significantly larger in erosions and restenotic lesions compared to stable plaques. In the same location a striking decrease of SMMHCs and a disappearance of smoothelin was observed in all situations.

Conclusions:  Intimal SMCs of erosions and restenotic lesions as well as, to a lesser extent, of stable plaques, exhibit a phenotypic profile suggesting that they have modulated into myofibroblasts. The high accumulation of α‐SMA‐positive cells in erosions correlates with the higher appearance of thrombotic complications in this situation compared to the two other situations. (Supported by the Swiss National Science Foundation grant No 32‐068034.02.)