Proapoptotic effect of control-released basic fibroblast growth factor on skin wound healing in a diabetic mouse model.

The ability of basic fibroblast growth factor (bFGF) to improve wound healing is attenuated by its short half-life in free form. This study aimed to enhance skin wound healing in a diabetes mouse model while concomitantly decreasing scar formation using control-released bFGF together with acidic gelatin hydrogel microspheres (AGHMs). Bilateral full-thickness wounds (10 mm in diameter) were made on the backs of db/db mice. Forty-five mice were divided into three groups, and the base of the wound under the panniculus carnosus and the wound periphery were injected with phosphate-buffered saline (300 µL) containing (1) control-released bFGF (50 µg), (2) control-released bFGF (20 µg), or (3) AGHMs alone. The size of the wound area was recorded on each postoperative day (POD). Mice were sacrificed on postoperative day 4, 7, 10, 14, and 28, and skin wound specimens were obtained to assess the endothelium/angiogenesis index via cluster of differentiation 31 immunohistochemistry, the proliferation index via Ki-67 immunohistochemistry, and the myofibroblast and fibroblast apoptosis indices by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and alpha-smooth muscle actin or vimentin staining, respectively. Epithelialization rates and indices of proliferation and myofibroblast/fibroblast apoptosis were higher in the bFGF groups than in the AGHM group, mainly within 2 weeks of injury. No dose-effect relationship was found for control-released bFGF, although the actions of 50 µg bFGF seemed to last longer than those of 20 µg bFGF. Therefore, control-released bFGF may accelerate diabetic skin wound healing and induce myofibroblast/fibroblast apoptosis, thereby reducing scar formation.

The Clinical Application of Preexpanded and Prefabricated Super-Thin Skin Perforator Flap for Reconstruction of Post-Burn Neck Contracture.

OBJECTIVE: Based on our previous animal study, we applied the "bridging effect" to the neighboring axial flap through preexpansion and prefabrication of a skin perforator flap as a new method to reconstruct a large skin defect after release of severe neck burn scar contracture. METHODS: Twelve patients suffering from severe post-burn cervical contractures underwent reconstruction of large skin defects after surgical release of severe scar contractures with preexpanded and prefabricated super-thin skin perforator flaps supplied primarily by a number of perforators via the "bridging effect" from the branches of the adjacent arteries as 2-stage procedures. During the first-stage operation, 2 tissue expanders were placed accordingly, and this was followed by a subsequent second-stage procedure where an expanded super-thin skin perforator flap was transposed to reconstruct a large neck skin defect. Follow-up was between 6 months and 3 years in this series. RESULTS: All super-thin skin perforator flaps survived in this series with primary healing except one with a distal flap necrosis that was treated with a subsequent skin graft. All patients have had a good contour with improved range of motion in the neck. CONCLUSIONS: The preexpansion and prefabrication of a super-thin skin perforator flap can possibly improve the anastomoses between neighboring subdermal vascular plexuses and extend the supplying area of these vessels to the flap. This method may provide a favorable super-thin skin flap that can be used for reconstruction of large neck defects after release of post-burn cervical scar contracture as demonstrated in this case series.

Are keloid and hypertrophic scar different forms of the same disorder? A fibroproliferative skin disorder hypothesis based on keloid findings.

Hypertrophic scars (HSs) and keloids are commonly seen as two different diseases by both clinicians and pathologists. However, as supported by histological evidence showing they share increased numbers of fibroblasts and accumulate collagen products, HS and keloid might be different forms of the same pathological entity, rather than separate conditions. To test this hypothesis, keloids from patients who underwent scar excisions (n = 20) in Nippon Medical School from 2005 to 2010 were examined histologically. The proportion and distribution of cellular and matrix collagen components were evaluated at the centre and periphery of each sample. In keloid samples, coexistence of hyalinised collagen, which is the most important pathognomonic characteristic of a keloid and dermal nodules that are considered to be characteristic of HS, was found. Moreover, hyalinised fibres appeared to initiate from the corner of the dermal nodules. Key features of inflammation such as microvessels, fibroblasts and inflammatory cells all decreased gradually from the periphery to the centre of keloids, indicative of reduced inflammation in the centre. Thus, we hypothesise that HS and keloid can be considered as successive stages of the same fibroproliferative skin disorder, with differing degrees of inflammation that might be affected by genetic predisposition.

Periodontal tissue regeneration by combined implantation of adipose tissue-derived stem cells and platelet-rich plasma in a canine model.

BACKGROUND AIMS: One goal of periodontal therapy is to regenerate periodontal tissues. Stem cells, growth factors and scaffolds and biomaterials are vital for the restoration of the architecture and function of complex tissues. Adipose tissue-derived stem cells (ASCs) are an ideal population of stem cells for practical regenerative medicine. In addition, platelet-rich plasma (PRP) can be useful for its ability to stimulate tissue regeneration. PRP contains various growth factors and may be useful as a cell carrier in stem cell therapies. The purpose of this study was to determine whether a mixture of ASCs and PRP promoted periodontal tissue regeneration in a canine model. METHODS: Autologous ASCs and PRP were implanted into areas with periodontal tissue defects. Periodontal tissue defects that received PRP alone or non-implantation were also examined. Histologic, immunohistologic and x-ray studies were performed 1 or 2 months after implantation. The amount of newly formed bone and the scale of newly formed cementum in the region of the periodontal tissue defect were analyzed on tissue sections. RESULTS: The areas of newly formed bone and cementum were greater 2 months after implantation of ASCs and PRP than at 1 month after implantation, and the radiopacity in the region of the periodontal tissue defect increased markedly by 2 months after implantation. The ASCs and PRP group exhibited periodontal tissue with the correct architecture, including alveolar bone, cementum-like structures and periodontal ligament-like structures, by 2 months after implantation. CONCLUSIONS: These findings suggest that a combination of autologous ASCs and PRP promotes periodontal tissue regeneration that develops the appropriate architecture for this complex tissue.

Identification and characterization of Wnt signaling pathway in keloid pathogenesis.

Keloid is characterized by fibroblastic cell proliferation and abundant collagen synthesis. Numerous studies have shown that the Wingless type (Wnt) signaling pathways play key roles in various cellular functions including proliferation, differentiation, survival, apoptosis and migration. The aim of this study was to clarify the role of Wnt signaling pathway in keloid pathogenesis. Primary fibroblast cultures and tissue samples from keloid and normal appearing dermis were used. The expression of Wnt family members, frizzled (FZD)4 receptor, receptor tyrosine kinase-like orphan receptor (ROR)2 and the Wnt signaling downstream targets, glycogen synthase kinase (GSK)3-ß and ß-catenin were assessed using semi-quantitative RT-PCR, Western blot, or immunohistochemical methods. Of the Wnt family members, Wnt5a mRNA and protein levels were elevated in keloid fibroblasts (KF) as compared to normal fibroblasts (NF). A higher expression of ß-catenin protein was also found in KF. No detectable levels of FZD4 receptor and ROR2 proteins were observed in both NF and KF. Functional analysis showed that treatment of NF and KF with recombinant Wnt5a peptide resulted in an increase in protein levels of total ß-catenin and phosphorylated ß-catenin at Ser33/37/Thr 41 but no significant change in phosphorylated ß-catenin at Ser45/Thr 41 positions. In addition, the expression of total GSK3-ß protein was not affected but its phosphorylated/inactivated form was increased in NF and KF. Our findings highlight a potential role for a Wnt/ß-catenin canonical signaling pathway triggered by Wnt5a in keloid pathogenesis thereby providing a new molecular target for therapeutic modulations.

Preclinical efficacy of slow-release bFGF in ischemia-reperfusion injury in a Dorsal Island skin flap model.

The effect of slow-release basic fibroblast growth factor (bFGF) on ischemia-reperfusion injury was examined using an island skin flap model in rats. Paired rectangular island skin flaps were elevated on the dorsum of 30 Fischer rats. The flaps were subjected to 6 hours of ischemia. Before reperfusion the flaps were injected with acidic gelatin hydrogel microspheres + phosphate-buffered saline (PBS) (group I), 20 µg slow-release bFGF + PBS (group II), 50 µg slow-release bFGF + PBS (group III), and 150 µg slow-release bFGF + PBS (group IV). The mean percent flap survival area and the average number of vessels detected by microangiography were significantly higher in group IV (p < 0.05) than in groups I, II, and III. The immunohistochemical staining for vasculogenic growth factors was quantitatively higher in group IV (p < 0.01). In conclusion, slow-release bFGF prevents ischemia-reperfusion injury by upregulating the secretion of vasculogenic growth factors.

The relationship between skin stretching/contraction and pathologic scarring: the important role of mechanical forces in keloid generation.

Keloids tend to occur on highly mobile sites with high tension. This study was designed to determine whether body surface areas exposed to large strain during normal activities correlate with areas that show high rates of keloid generation after wounding. Eight adult Japanese volunteers were enrolled to study the skin stretching/contraction rates of nine different body sites. Skin stretching/contraction was measured by marking eight points on each region and measuring the change in location of the marked points after typical movements. The distribution of 1,500 keloids on 483 Japanese patients was mapped. The parietal region and anterior lower leg were associated with the least stretching/contraction, while the suprapubic region had the highest stretching/contraction rate. With regard to keloid distribution, there were 733 on the anterior chest region (48.9%) and 403 on the scapular regions (26.9%). No keloids were reported on the scalp or anterior lower leg. Because these sites are rarely subjected to skin stretching/contraction, it appears that mechanical force is an important trigger that drives keloid generation even in patients who are genetically predisposed to keloids. Thus, mechanotransduction studies are useful for developing clinical approaches that reduce the skin tension around wounds or scars for the prevention and treatment of not only keloids but also hypertrophic scars.

Small-wave incision method for linear hypertrophic scar reconstruction: a parallel-group randomized controlled study.

BACKGROUND: The usual hypertrophic scar reconstruction methods such as Z- and W-plasties can leave noticeable scars and involve excessive normal skin excision, particularly in long linear hypertrophic scars longer than 10 cm. Thus, we invented and applied the small-wave incision method for patients with linear hypertrophic scars. METHODS: A total of 40 patients with linear hypertrophic scars were included in this study. The patients were randomly assigned to the linear incision group (n = 20) or the small-wave incision group (n = 20). All scars were mildly hypertrophic, longer than 10 cm, and located in the lower abdominal/suprapubic region. They occurred after Cesarean section or gynecological or abdominal surgery. Patient age and sex, the cause of the scar, and the preoperative and postoperative sizes of the scar were recorded. Postoperative scar size and recurrence were evaluated for 18 months. Mathematical comparisons were also made to multiple linear incision, Z-plasty, planimetric Z-plasty, and W-plasty. RESULTS: Postoperative recurrence was 40 and 15% in linear and small-wave groups, respectively (P = 0.77). The main risk factor for recurrence was postoperative size (P = 0.043). Mathematical comparisons revealed that the small-wave method can achieve the same release of tension with the least normal skin excision while making the scar irregular via an accordion effect. CONCLUSION: The small-wave method can meet both the functional and the cosmetic requirements of long linear hypertrophic scar reconstruction while reducing complication risks.

Chest wall reconstruction of severe mediastinitis with intercostal artery-based pedicled vertical rectus abdominis muscle flap with oblique-designed skin pedicle.

Recurrent poststernotomy mediastinitis has significant morbidity and mortality. Reconstructive treatment begins with pectoral muscle or omental flaps. When these options are unavailable or inadequate, surgeons resort to internal mammary artery-based vertical rectus abdominis muscle flap. If the internal mammary artery is harvested for coronary artery bypass grafting, surgeons are reluctant to use the muscle for pedicled flap because of the elevated risks. However, recent reports suggest that if enough time passes for intercostal artery collaterals to develop, they would support the viability of the flap. Moreover, recent improvements in defining flap microvasculature and proposed surgical techniques have enabled us to further refine the procedure. Although it appears that the intercostal artery-based pedicled vertical rectus abdominis muscle flap with oblique-designed skin pedicle is safe and effective for chest wall reconstruction, potential for failure remains elevated until sample size accumulates.

Clinical applications of perforator-based propeller flaps in upper limb soft tissue reconstruction.

PURPOSE: A propeller flap is an island flap that moves from one orientation to another by rotating around its vascular axis. The vascular axis is stationary, and flap movement is achieved by revolving on this axis. Early propeller flaps relied on a thick, subcutaneous pedicle to maintain vascularity, and this limited the flap rotation to 90°. With increasing awareness of the location and the vascular territory perfused by cutaneous perforators, it is now possible to design propeller flaps based on a single perforator, so-called "perforator-based propeller flaps." These flaps permit flap rotation up to 180°. We present the results of upper limb soft tissue reconstruction using perforator-based propeller flaps. We constructed a treatment strategy based on the location of the soft tissue defect and the perforator anatomy for expedient wound coverage in 1 stage. METHODS: All perforator-based propeller flaps derived from 3 institutions that were used for upper limb soft tissue reconstruction were retrospectively analyzed. The parameters studied included the size and location of the defect, the perforator that was used, the size and shape of the flap, the direction (ie, clockwise or counter-clockwise) of flap rotation, the degree of twisting of the perforator, the management of the donor site (ie, linear closure or skin grafting), and flap survival (recorded as the percentage of the flap area that survived). RESULTS: Twelve perforator-based propeller flaps were used to reconstruct upper limb soft tissue defects in 12 patients. Six different perforators were used as vascular pedicles. The donor defects of 11 flaps could be closed primarily. One flap was partially lost in a patient with electrical burns. CONCLUSIONS: Perforator-based propeller flaps provide a reliable option for covering small- to medium-size upper limb soft tissue defects.

Acellular dermal matrix seeded with adipose-derived stem cells as a subcutaneous implant.

BACKGROUND: Soft tissue augmentation using autologous materials usually is associated with high resorption rates. To obtain more durable soft tissue filler, acellular dermal matrix (ADM) was seeded with adipose-derived stem cells (ASCs) in this study. METHODS: For ADM preparation, split-thickness skin was obtained from the dorsum of two Fischer rats. Harvested skin was acellularized to obtain ADM and subsequently seeded in vitro with ASCs isolated from the same rats. Subcutaneous soft tissue augmentation was carried out in the dorsal area of 20 Fischer rats. The implant materials were ADM (group 1), ADM with ASCs (group 2), collagen type 1 gel (group 3), and collagen type 1 gel with ASCs (group 4). Each specimen was harvested after 8 weeks for quantitative evaluation of thickness, vascular density, and collagen content. RESULTS: Histologic analysis showed that ASCs were successfully seeded onto ADM. The thickness of the implanted material and the vascular density were highest 8 weeks postoperatively in group 2. The subpanniculus layer became significantly thicker in group 3 than in group 4. The collagen content was significantly higher in group 2 than in the other groups. CONCLUSION: These findings suggest that ADM seeded with ASCs forms the best implant material in terms of volume maintenance, vascular density, and collagen content.

Visual and pathologic analyses of keloid growth patterns.

Keloids grow and spread not only vertically but also horizontally, although hypertrophic scars do not grow beyond the boundaries of the original injury. Clinically, we have encountered keloids with regular and irregular (untypical) shapes. As the characteristics of the irregular growth patterns of keloids have not been studied yet, we analyzed the irregular growth of keloids both visually and pathologically.A total of 220 keloid specimens, each from a different patient, were surgically removed and used in this study. Through visual analysis, the preoperative shapes of these 220 keloids were classified into those with a regular shape (R group) and those with an irregular shape (IR group). Moreover, we distinguished between cases that had received keloidectomy previously and those that had not. We also determined whether the keloids were recurrent keloids or not. In both the R and IR groups, keloid specimens were studied histologically to examine for infection.In the R group, there were 156 cases (70.9%; 55 males and 101 females with a mean age of 33.68 years). Three patients (1.9%) had infection and 2 patients (1.3%) had undergone keloidectomy previously. In the IR group, there were 64 cases (29.1%; 24 males and 40 females with a mean age of 45.27 years). Thirty patients (46.9%) had infection and 24 patients (37.5%) had undergone keloidectomy previously. Statistically, the rates of infection and keloidectomy were significantly different between the R group and the IR group.Severe infection or operative history may be the cause of irregularly shaped keloids. Thus, in the absence of significant infection or a surgical history, the shape of keloids may be determined uniquely by skin tension.

Prefabrication of tissue engineered bone grafts: an experimental study.

The purpose of this study was to determine whether angiogenesis could successfully be induced into bone tissue that was engineered by cultured adipose-derived stem cells with porous beta-tricalcium phosphate and whether its biologic properties could be maintained by flap prefabrication technique.Adipose-derived stem cells with porous beta-tricalcium phosphate were implanted into the superficial inferior epigastric artery flap of the Fisher rats. After prefabrication for 8 weeks, the prefabricated flaps were elevated and the pedicles were clamped for 4 hours. The samples were harvested after 2 weeks for analyses.Angiogenesis was significantly increased in the prefabricated groups (P < 0.05). There was no significant difference between the prefabricated and nonprefabricated groups in terms of the osteogenic capacity (P > 0.5).The promising results obtained with prefabrication in tissue engineered bone grafts encourage the clinical application of this technology. Thus, prefabrication may be a useful technique in any engineered bone tissue transfer.

Effect of mesenchymal stem cells on skin graft to flap prefabrication: an experimental study.

Angiogenetic potential has been reported for bone marrow-derived stem cells (BSCs) and adipose-derived stem cells (ASCs). The superficial femoral artery, vein, and fascia were used as a vascular crane for prefabrication model of skin graft to flap. BSCs or ASCs were injected before the adaptation of the graft to the vascular crane depending on the group. The prefabricated grafts were then transferred to inguinal region in every 7 days to observe the viability. In experiment part I (n = 18), the critical time for the prefabrication was found to be 1 week. In experiment part II (n = 12), the control and experiment assays were performed on the same animal to support the data of the experiment part I. The viability of flaps was evaluated. The vascular density was higher in BSC, and ASC groups. The Vascular Endothelial Growth Factor immunohistochemical staining was quantified. Furthermore, mesenchymal stem cells could be helpful in any prefabrication procedure in which neovascularization is indispensable.

A case of von Willebrand disease discovered during treatment of a sacral pressure ulcer.

A sacral pressure ulcer developed in a patient hospitalized for cerebral infarction. Each time necrotic tissue was debrided from the ulcer, pressure hemostasis was necessary to stop the bleeding. As treatment continued, the pressure required to stop the bleeding caused the ulcer to worsen, leading to a downward spiral in the patient's condition. While trying to determine the cause of this problem, we discovered that the patient had von Willebrand disease. Medication controlled the bleeding, and the pressure ulcer began to heal at the same time. It was clear to us that conservative treatment would lead to a complete cure but that the healing process would take a long time and require continued administration of an expensive drug. We decided, therefore, to close the wound with a fasciocutaneous flap so that the patient could be quickly transferred to a rehabilitation hospital. About 1 month after surgery, epithelialization was complete, we were able to discontinue medication, and the patient was discharged. This experience demonstrates the importance of determining the cause of any deviation from the normal course of healing in pressure ulcers. It also indicates that the use of fasciocutaneous flaps, which involve little intraoperative bleeding in short surgeries, is appropriate in cases like this one.

Merkel cell carcinoma on the upper lip of a 100-year-old woman.

Merkel cell carcinoma (MCC) is a rare skin tumor that usually occurs on the head, neck, or extremities of elderly patients; it has a high incidence of local recurrence, regional lymph node metastasis, and subsequent distant metastasis. We report a MCC that developed rapidly on the left corner of the upper lip of a 100-year-old woman. An incisional skin biopsy was performed to confirm MCC. Computed tomography showed no metastasis. The tumor was widely excised with a margin of 1 cm. Immediate reconstruction with a reverse Estlander flap from the lower lip was performed under general anesthesia. Additional surgery was also performed under general anesthesia 2 weeks later to widen the patient's lips. The surgical results were satisfactory. The patient died of senile deterioration a year after hospitalization for long-term medical treatment, without any recurrence or metastasis of MCC. Despite the patient's age, we considered it necessary to resect the tumor widely because of its rapid growth. The tumor margin was 1 cm. No radiotherapy was performed, but we believe that surgery alone was effective in allowing this patient to live an additional year without recurrence or metastasis. To the best of our knowledge, this patient is the oldest person with MCC yet described.

Preoperative administration of Planovar® in two cases of abdominal wall endometriosis after cesarean section.

We describe 2 cases of abdominal wall endometriosis, a condition that usually occurs in surgical scars from previous cesarean sections and presents as a mass with cyclic pain and swelling. Wide local excision with clear margins seems to be the only effective treatment. However, the mass in our 2 cases changed in size with menstruation, and palpating the masses was sometimes difficult. Therefore, we administered an oral contraceptive containing a combination of estrogen and progesterone (Planovar®) preoperatively to each patient so that the endometrial mass could be reliably palpated on the day of operation. The mass was excised cleanly in each case.

Microscope-assisted reduction clitoroplasty used to treat two patients with clitoromegaly.

We report 2 cases of clitoromegaly, 1 in a patient with true hermaphroditism, and the other in a patient with adrenogenital syndrome. Both were treated surgically with reduction clitoroplasty. There are 3 different clitoroplasty procedures: clitorectomy, clitoral recession, and reduction clitoroplasty. Reduction clitoroplasty with preservation of the neurovascular bundle is considered superior in terms of formation of the external genitals and sensation. However, the disadvantages are that detachment of the neurovascular bundle from the clitoral shaft is difficult and that there is a high possibility of sensory and blood flow disorders in the clitoris. In an attempt to achieve safe and reliable surgical manipulation, we used a surgical microscope (OPMI 6-SDFC, Carl Zeiss Surgical GmbH, magnification x8) to detach the neurovascular bundle from the clitoral shaft in our 2 patients. Our impression is that our efforts were extremely effective. Furthermore, our experience leads us to believe that the procedure for neurovascular bundle detachment required in reduction clitoroplasty is not particularly difficult if performed with a surgical microscope by a plastic surgeon who regularly performs microsurgery. Because the procedure can be performed simply and safely, we believe that reduction clitoroplasty with preservation of the neurovascular bundle is the best overall of the 3 clitoroplasty procedures.

Fat grafting to the breast and adipose-derived stem cells: recent scientific consensus and controversy.

Recent technical advances in fat grafting and the development of surgical devices such as liposuction cannulae have made fat grafting a relatively safe and effective procedure. However, new guidelines issued by the American Society of Plastic Surgeons in 2009 announced that fat grafting to the breast is not a strongly recommended procedure, as there are limited scientific data on the safety and efficacy of this particular type of fat transfer. Recent progress by several groups has revealed that multipotent adult stem cells are present in human adipose tissue. This cell population, termed adipose-derived stem cells (ADSC), represents a promising approach to future cell-based therapies, such as tissue engineering and regeneration. In fact, several reports have shown that ADSC play a pivotal role in graft survival through both adipogenesis and angiogenesis. Although tissue augmentation by fat grafting does have several advantages in that it is a noninvasive procedure and results in minimal scarring, it is essential that such a procedure be supported by evidence-based medicine and that further basic scientific and clinical research is conducted to ensure that fat grafting is a safe and effective procedure.

Complications after self-injection of hyaluronic acid and phosphatidylcholine for aesthetic purposes.

Various types of dermal fillers have been developed for soft tissue augmentation, including hyaluronic acid products, which have been approved by regulatory agencies in Europe, the United States, and elsewhere. Phosphatidylcholine (PPC) injection has attracted attention for its application in the treatment of cellulite, weight loss, and skin rejuvenation. Because the public can now buy PPC and similar products from various online pharmacy websites without the involvement of a clinician, there is potential for misuse. The authors discuss two cases of complications experienced by patients after self-injection of hyaluronic acid and PPC for aesthetic purposes.