Ryuzaburo Tanino

Induction of vascular endothelial growth factor by fibrin as a dermal substrate for cultured skin substitute.

In the initial phase of wound healing, endogenous fibrin clots are known to form a provisional matrix and to promote angiogenesis. Growth factors such as vascular endothelial growth factor (VEGF) increase in wounds to stimulate angiogenesis. However, it remains unknown whether VEGF is induced when fibrin is used as a dermal substrate for cultured skin substitutes. The authors investigated the effect of fibrin gel as a dermal substrate for a cultured skin substitute, using human keratinocytes and dermal fibroblasts. A collagen-cultured skin substitute was also examined for comparison. VEGF in the culture supernatant in both types was measured by enzyme-linked immunosorbent assay, and VEGF mRNA was determined semiquantitatively by reverse-transcriptase polymerase chain reaction after 2 days of incubation. Experiments were performed using 12 cultured skin substitutes: four for histologic examination before transplantation, four for VEGF assay in vitro, and four for the transplantation to athymic mice. Three independent experiments were performed for each step. VEGF concentration in the fibrin-cultured supernatant was 84.3 +/- 11.8 pg/ml, whereas it was 27.8 +/- 4.68 pg/ml in the case of the collagen substrate. The relative levels of VEGF mRNA were 1.088 +/- 0.100 and 0.698 +/- 0.226, respectively. In in vivo transplantation, the fibrin-type cultured skin substitute showed an excellent take on the wound bed, and a normally proliferating keratinocyte layer with emergence of vascular endothelial cells in the transplanted floor was seen 3 days after transplantation. Vascular endothelial cells, which were identified using alkaline phosphatase stain, were significantly increased in the fibrin-type cultured skin substitute. The use of fibrin as a dermal substrate for cultured skin substitute increases the secretion of VEGF, improves regeneration of mature epidermal structure after in vivo transplantation, and promotes the migration of vascular endothelial cells.

The influence of different types of hard-palate closure in two-stage palatoplasty on maxillary growth: cephalometric analyses and long-term follow-up.

Using cephalometric analysis we investigated the influence on maxillary growth of two different types of hard-palate closure in two-stage palatoplasty. In 12 patients with complete unilateral cleft lip, alveolus, and palate, the lip and soft-palate were closed between 3 and 7 months of age. These 12 patients were then assigned to two groups of 6. In one group the hard palate was closed at 1 year 5 to 11 months of age by a vomer flap with a skin graft (VF group, Osadas two-stage palatoplasty) and in the other group it was closed by the mucoperiosteal pushback procedure (PB group). Sella-nasion-point A (SNA) in the VF group at 3 to 4 and after 10 years of age were within normal range and significantly larger than in the PB group. Two patients in the PB group required orthognatic surgery to obtain normal occlusion and a well-balanced profile. We concluded that in two-stage palatoplasty better maxillary growth can be obtained using the vomer flap method than using the pushback procedure.

A microangiographic technique using synchrotron radiation to visualize dermal circulation in vivo

&NA; Conventional angiography cannot resolve dermal small vessels with a diameter of 200 &mgr;m or less. In vitro microangiography is currently characterized by better spatial resolution than conventional angiography but does not allow visualization of the blood stream in dermal vessels in vivo. In this study, we introduce a novel synchrotron radiation microangiographic system for visualizing the structure of and blood flow in dermal microvessels in vivo repeatedly. We used monochromatic synchrotron radiation with an energy just above the k‐edge of iodine (33.3 keV) as an x‐ray and a high‐definition television camera system with a high‐sensitivity image pick‐up tube for detection. The 33.3‐keV monochromatic synchrotron radiation allows detection of a small amount of iodine, and the high‐definition television camera system can resolve small vessels with high‐spatial resolution and no loss of sensitivity. We performed synchrotron radiation angiography of superficial inferior epigastric arteries and their branches in nine rats, and of the caudal artery in 14 rats, and compared angiographic images taken by the current system with those taken by a conventional angiographic system in seven rats. With this new microangiographic technique, we could visualize small dermal vessels with a diameter as low as 50 &mgr;m. In addition, repeated angiograms at baseline and under increasing body temperature could be obtained. This new microangiographic approach is expected to be very useful for the assessment of dermal circulation in patients. (Plast. Reconstr. Surg. 102: 1128, 1998.)

Attempt to obtain greater dermal depth of vascular injury using dye-enhanced laser technique: A new approach

Although pulsed dye laser has been successfully used in the treatment of portwine stains, a number of patients had incomplete clearance because the depth of penetration by the pulsed dye laser was inadequate. This study was performed to establish the greater penetration depth of vascular injury using a dye‐enhanced laser technique.

Surgical treatment of facial steatocystoma multiplex

Facial steatocystoma multiplex is a rare variant of steatocystoma multiplex, which is characterized by multiple disfiguring cysts on the face and is inherited in an autosomal dominant fashion. Noninvasive treatments are only temporarily effective, but surgery of multiple lesions on the face has a risk of major complications. A 31-year-old man presented with papules distributed over his face. We used a flap method and mini-incisional capsulectomy under general anesthesia. We performed needle aspiration and CO2 laser puncture and squeezing on the man in the outpatient clinic when symptoms recurred. There have been several reports suggesting various options for the management of steatocystoma. However, long-term management remains difficult and there is no definite technique to treat facial steatocystoma multiplex. We think that combined use of these surgical methods should be applied with individual evaluation. Size, location, hardness, mobility, depth, and previous scars should be noted, and the relevant methods should be selected for each cyst.

Experience of the treatment of melanin deposit benign cutaneous pigmented lesions with PLDL-1

Benign cutaneous pigmented lesions have been treated by many conventinal methods,such as dry ice therapy,dermabrasion or laser therapy.But such conventinal methods sometimes resulted in hyperpigmentation or scarformation. A 510nm dye laser with pulse duration of 375nsec (PLDL-1) was used to treat 3lpatients with benign cutaneous pigmented lesions. Skin lesions were nevi spilus,senile lentigines,nevocellular nevi,and traumatic tattoo. 74%cases were effective by this laser treatment,and no hyperpigmentation or scarformation was seen in all cases. This laser was very easy to treat and was thought to be useful for superficial cutaneous pigmented lesion treatment.

Possibility of Treating Hyperpigmented Skin Lesions Using the Nd: YAG Laser

The authors have developed a ruby laser system having a device to provide uniform output, the “Kaleidoscope” handpiece. This equipment can be used for the treatment of a hyperpigmented skin lesions, such as nevus spilus and nevus cell nevus. As a result, acceptable clinical results have been obtained*. However, the ruby laser was not effective for the treatment of deeply located intra-dermal hyper-melanoses.