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Acute hand edema often results in loss of fine hand motor activities, especially without appropriate care. There is still no reliable and easy to use method to measure hand edema. In this study, we tested a handheld three-dimensional (3D) scanner on plastic male and female hand models using a whole hand measuring method (WM) and a modified method (MM) which excluded fingers. We evaluated the intra-rater reliability and inter-rater reliability and compared the measured volumes to computed tomography (CT) findings. Statistical analysis showed that the 3D scan method was valid and reliable for both WM and MM methods. In WM, intra-rater and inter-rater reliability were 0.97 and 0.84, with 95% confidence interval (CI) of 0.87-1.00 and 0.61-0.94, respectively. In MM, intra-rater and inter-rater reliability were 0.96 and 0.83, with 95% CI of 0.84-1.00 and 0.61-0.94, respectively. In comparison to the CT, the differences between 3D scan and CT in the male model volumes were 30.35 ± 2.70 cm

Stem cell transplantation is expected to be an effective treatment for intractable skin ulcers by promoting angiogenesis; however, it is challenging to quickly realize a sufficient bloodstream for the ulcers. For this treatment, sheet-like materials with monolayer cells such as cell sheets have been investigated. However, they have a limitation of cell number that can be transplanted at one time due to the two-dimensional, monolayer cell structure, and sufficient secretion of growth factors cannot be expected. In this regard, cellular aggregates, such as spheroids, can reproduce three-dimensional cell-cell interactions that cause biological functions of living tissues more representative than monolayer cells, which is important to achieving efficient secretion of growth factors. In this study, we focused on free-standing porous polymer ultrathin films ("porous nanosheets") comprising poly(d,l-lactic acid) (PDLLA) and succeeded in developing a spheroid-covered nanosheet, on which more than 1000 spheroids from adipose-tissue derived stem cells (ASCs) were loaded. The porous structure with an average pore diameter of 4 μm allowed for facile filtration and carrying spheroids on the nanosheet, as well as sufficient oxygen and nutrients inflow to the cells. The spheroid-covered nanosheet achieved homogeneous transference of spheroids to a whole skin defect in diabetic model mice. Given the continuous release of vascular endothelial growth factor (VEGF) from the spheroids, the transplanted spheroids promoted healing with more accelerated angiogenesis than a nanosheet with a monolayer of cells. The spheroid-covered nanosheet may be a new regenerative material for promoting intractable skin ulcer healing.

The short survival time of transplanted adipose-derived mesenchymal stem cells (ASCs) is a problem for skin wound healing. Transplantation after the formation of cellular spheroids has been investigated as a promising method for prolonging cellular survival. However, there have been technical restrictions for transplantation of spheroids in clinical practice. Here, we show an effective method for transplantation of ASC spheroids onto skin wounds in order to efficiently cure refractory ulcers. To assist anchoring of spheroids onto skin wounds, we used a 120-nm-thick free-standing film (nanosheet) that has a highly adhesive property. Bioluminescence imaging showed that ASC spheroids carried by the nanosheet survived for 14 days, which is about two-times longer than that previously reported. Wounds treated with a nanosheet carrying ASC spheroids were 4-times smaller than untreated wounds on day 14. This method for transplantation of spheroids could be applied to cell therapy for various refractory skin wounds.

Objectives To verify the utility of triangular extension of a hinge flap in buccal mucosal staged urethroplasty to resolve stomal stenosis after the first stage and ultimately prevent restenosis. Methods A total of 23 patients (triangular extension group) were studied in 2013-2019. In the first stage, buccal mucosa was transplanted, and an extended triangle portion of the mucosa was placed beside the proximal and/or distal stoma that was created when the stricture segment of the urethra was resected. In the second stage, during tubularization of the urethral plate, an incision was made at the stoma to increase the caliber to which the triangular extension was inserted. The procedure was considered successful when a 17-Fr flexible cystoscope passed through the reconstructed urethra at 6 months after the second-stage urethroplasty and no additional surgery or bougie dilation required. The clinical course of the triangular extension group was compared with 24 patients who underwent conventional staged urethroplasty (control group). Results In total, 20 patients from each group underwent second-stage surgery. No patients in the triangular extension group required additional revision surgery because of stomal stenosis after first-stage surgery, whereas five (20%) control patients did. Urethroplasty was successful in 19 patients (95%) in the triangular extension group and in 19 patients (95%) in the control group. Uroflowmetry after the second-stage surgery indicated that the mean maximum urinary flow rate was 21.5 and 15.8 mL/s after triangular extension and the control procedure, respectively (P = 0.027). Conclusions The triangular extension technique reduces the need for revision surgery and prevents postoperative restenosis.

The local flap method is a minimally invasive, quick, and common technique of closing skin defects. Several methods using multiple local flaps are employed for the purpose of closing relatively large defects, and minimizing the resultant scar and deformity. We present a combined local flap method using a banner flap with an adjacent rotation flap (B-R flap) with the potential for closing circular defects. A retrospective analysis was performed in 15 patients treated with the B-R flap for circular skin defects over a 2-year period. The sizes of the flap and the healthy skin excision area were evaluated using image analysis software, and were compared to a model of a conventional rotation flap that would be necessary to close the same defect. All flaps were successfully engrafted. The B-R flap required a 39.1% smaller rotation flap and an 85.9% smaller area of healthy skin compared to the conventional rotation flap technique. There was one adverse event, partial epidermal necrosis that was conservatively treated and healed. The B-R flap is comparatively less invasive; it allows a smaller area of healthy skin to be excised. It can be a useful option for closing a circular defect.

Since the majority of a tendon’s dry weight is collagen fibers, tendon healing consists mainly of collagen repair and observing three-dimensional networks of collagen fibers with scanning electron microscopy (SEM) is optimal for investigating this process. In this report, a cell-maceration/SEM method was used to investigate extrasynovial tendon (unwrapped tendon in synovial tissue such as the tendon sheath) healing of an injured Achilles tendon in a rat model. In addition, since mechanical stimulation is important for tendon healing, a novel, tensionless, rat lower leg tendon injury model was established and verified by visualizing the structural change of collagen fibers under tensionless conditions by SEM. This new model was created by transplanting the leg of a rat with a tendon laceration to the back, removing mechanical stimulation. We then compared the process of tendon healing with and without tension using SEM. Under tension, collagen at the tendon stump shows axial alignment and repair that subsequently demarcates the paratenon (connective tissue on the surface of an extrasynovial tendon) border. In contrast, under tensionless conditions, the collagen remains randomly arranged. Our findings demonstrate that mechanical stimulation contributes to axial arrangement and reinforces the importance of tendon tension in wound healing.

We report a 29-year-old man with blepharochalasis and multiple chemical sensitivity (MCS). Although standard blepharoplasty with aponeurotic fixation was performed, palpebral ptosis recurred after 3 months. Eyelid function and appearance improved after additional operations. A stepwise surgical approach is appropriate in patients with blepharochalasis and MCS.

A 72-year-old man presented with an erythematous, painful, swollen, and blistering left hand associated with a systemic fever. The patient was diagnosed with microscopic polyangiitis and was receiving steroid therapy from a year before the incident. Based on a clinical diagnosis of necrotizing fasciitis, emergency surgery was performed within 2 days after the onset. β-Hemolytic Streptococcus group A was isolated from a culture of the blood and wound. Radical debridement and high-dose penicillin and clindamycin therapy successfully saved the patient's life and affected limb except for the second finger on his left hand, which was completely necrotic. However, the function of the left hand was seriously decreased and did not recover. The important point to note in this case was the preexisting vasculitis neuropathy due to microscopic polyangiitis. The severe postoperative dysfunction of the hand was considered to be due to ischemic neuropathy that was aggravated by compartmental syndrome and microvascular thrombosis. In conclusion, necrotizing fasciitis of an extremity with underlying vasculitis neuropathy can lead to a poor functional prognosis of the limb.

Background Chest wall necrosis can manifest as a late effect of radiation therapy for breast cancer. Only two cases of fistulas communicating with the respiratory tract as a result of radiation-induced necrosis of the lungs or bronchi have been reported. To the best of our knowledge, we report the first case of a pulmonary cutaneous fistula arising as a late effect of radiation therapy for breast cancer, which was successfully repaired using a free omental graft. Case presentation A 64-year-old woman underwent Halsted surgery and postoperative radiation therapy for breast cancer 25 years earlier. One year before visiting our hospital, she developed a fistula and bleeding in her left clavicular region, which was expanding. On initial examination, a 6-cm-wide skin defect was observed in the left clavicular region and the clavicle appeared sequestrated. Computed tomography revealed part of the first to third left ribs, part of the left clavicle, the subclavian artery, and the brachial plexus to be missing. Several rounds of debridement revealed approximately 10 bronchial stumps on the surface of the collapsed lung, from which exhaled air and sputum were effusing. Surgery was performed to implant a free omental flap with vascular anastomosis and a skin graft in the neck region, and the pulmonary cutaneous fistula was closed. Two years after surgery, emphysema remained inside the omentum, which spontaneously resolved by the 3rd postoperative year. Conclusions Various treatment options are conceivable for the repair of pulmonary cutaneous and bronchocutaneous fistulas induced by radiation damage (e.g., free tissue grafts and endoscopic bronchial occlusion); however, these are rarely reported, and the most reliable method thus remains unclear. Positive outcomes in our case indicate that implanting a free omental graft may be effective. Furthermore, spontaneous healing can be expected for the residual emphysema inside the omentum.

Highlights • Nodular fasciitis is difficult to distinguish from sarcoma. • Clinical and pathological findings may not correspond, as in our case. • If sarcoma cannot be excluded, treatment should consider the risk of malignancy., Introduction Nodular fasciitis is a reactive proliferative lesion. It is often difficult to distinguish from sarcoma, such as malignant peripheral nerve sheath tumor (MPNST). Therefore, both pathological findings and clinical features must be considered. Presentation of case A 75-year-old man presented with a 1 × 1 cm mass on his left forearm that had enlarged rapidly without specific symptoms. Magnetic resonance imaging revealed a mass adjacent to brachioradialis. The lesion showed iso-intensity to muscle on T1-weighted images and hyperintensity on T2-weighted images. We made a clinical diagnosis of nodular fasciitis. Under observation, the mass gradually decreased in size and disappeared after two months. Four years later, another lesion appeared at the same site. We performed total excisional biopsy and histopathological examination indicated low grade MPNST or recurrent nodular fasciitis. Since malignancy could not be excluded, wide resection was performed. At 4 years postoperatively, there has been no recurrence. Discussion In this patient, nodular fasciitis of the right forearm was diagnosed clinically and showed spontaneous regression. However, recurrence was noted after four years. While the clinical features suggested recurrent nodular fasciitis, pathological findings indicated the possibility of low grade MPNST. Conclusion When it is difficult to determine whether a lesion such as nodular fasciitis is benign or malignant, the patient should be managed by considering the possibility of malignancy.

Many approaches have been reported to repair soft-tissue defects of the hand using dorsal metacarpal artery flaps. Use of a perforator-based propeller flap from the first intermetacarpal space to the dorsum of the hand has been described. The aim of this study was to confirm the functional anatomy of a first dorsal metacarpal artery (FDMA) perforator flap. Twenty-nine fixed cadaveric hands were dissected to determine the origin, course, and branches of the FDMA. Clinically, five cases of soft tissue defects of the hand underwent reconstructive surgery using an FDMA perforator-based propeller flap. The FDMA was found in 27 specimens (93%). The ulnar branch of the FDMA always supplied the cutaneous perforator (mean ± SD, 4.3 ± 1.6), and the most distal cutaneous perforating branch was found along the metacarpal long axis within 25 mm of the tip of the metacarpal head with high frequency (28/29, 97%). In the two hands that had aplasia of the FDMA, well-developed perforators arose directly from the radial artery and advanced to the metacarpal head. Seven hands (24%) had perforators arising from the palmar arterial system, penetrating through or passing close by the second metacarpal bone. In clinical application, all the flaps survived completely without major complications. The FDMA perforator-based propeller flap is minimally invasive and technically simple. It is expected to be a new option for hand reconstruction.

This is the first report of multiple parosteal lipomas as a late complication of aesthetic procedures. A 70-year-old woman presented with multiple frontal nodules that had grown slowly over 5 years. She underwent forehead-lifting surgery 30 years ago and botulinum toxin injection 3 years ago. Computed tomography showed six low-density nodules measuring 10 to 20 mm just above the frontal bone. The tumors were resected via a parietal coronal incision. Histopathological findings revealed parosteal lipoma with foreign bodies. At 1 year postoperatively, there has been no recurrence or complications. The multiple lesions corresponded to the sites of the aesthetic procedures. The mesenchymal cells in the periosteum seemed to be stimulated by cytokines released from a postoperative hematoma or damaged periosteum, resulting in tumorigenesis. When performing aesthetic procedures in the face, care should be taken to avoid unnecessary injury to the periosteum, given the possibility of tumor development.Level of evidence: Level V, case report.

Background Weakly acidic hypochlorous acid (HOCl), a major inorganic bactericidal compound of innate immunity, is effective against a broad range of microorganisms. However, HOCl has never been described as a pharmaceutical drug for treatment of infected wounds. Here, we describe a study using HOCl solution, a reagent that has potential use in a clinical situation such as prevention and treatment of infection in chronic wounds. Methods HOCl solution (200 ppm, pH 6.5) was prepared by the in vitro addition of sodium hypochlorite to sterile water or saline at pH 6.5. The resulting solutions were used to evaluate in vitro cytotoxic and bacteriostatic properties against Pseudomonas aeruginosa. Furthermore, the bacteriostatic effect of the HOCl solutions was evaluated in vivo using P. aeruginosa-infected wounds on db/db diabetic mice. Results The HOCl solution exhibited concentration-dependent in vitro bacteriostatic activity against P. aeruginosa; HOCl in phosphate-buffered saline (PBS) also displayed in vitro cytotoxicity to human fibroblasts. Similarly, the cleansing of P. aeruginosa-infected wounds on db/db diabetic mice with HOCl solution yielded significantly decreased in vivo bacterial cell counts and there was no difference in granulation formation compared to cleansing with pure water. Conclusion Twelve days of once-daily cleansing of P. aeruginosa-infected wounds on db/db diabetic mice with HOCl solution resulted in significantly decreased P. aeruginosa bioburden, albeit with a minor delay in wound repair.

Stem cell transplantation is expected to be an effective early-phase treatment for deep burn injuries and intractable ulcers. Localizing and proliferating stem cells on the lesion utilizing engineered scaffolds is important for this treatment. In this study, we demonstrated in situ transplantation of adipose-tissue derived stem cells (ASCs) organized on free-standing porous polymer ultrathin films (referred to as "porous nanosheets") to a skin defect model in diabetic mice. Porous nanosheets were prepared by a combination of micro-gravure coating with macrophase separation of poly(d,l-lactic acid) and polystyrene under a roll-to-roll process and solvent etching process with cyclohexane. The permeable structure of porous nanosheets (thickness of 150 nm, average pore diameter of 4 μm) allowed for proliferation of ASCs and also provided sufficient nutrient inflow into multilayered ASC constructs. Then, transplantation of a trilayered ASC-laden porous nanosheet achieved homogeneous transference of ASCs onto the skin lesion. Transplanted ASCs contributed to wound healing in a dorsal skin defect model in diabetic mice. Thus, cell transplantation using porous nanosheets will be a new method for promoting wound healing in diabetic and other kinds of refractory ulcers. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1363-1371, 2019.

Background Static reconstruction surgery that tightens the tension of the inferior tarsus, thereby raising the lax lower eyelid, is a common treatment for paralytic ectropion of the lower eyelid. We present one such operative procedure, in which an orbicularis oculi muscle flap was used. Methods The surgical technique involves partial resection of the tarsus and the skin, as well as a superior-based orbicularis oculi muscle flap that is sutured to the firm tissue present on the Whitnall tubercle. The muscle flap is approximately 7 mm in width and 15 mm in length, with a superior pedicle that is attached to the tarsus at the medial point of the resected tarsus. The procedure results in contact between the ocular surface and the lower eyelid. Results The procedure was performed in 11 patients with lower eyelid ectropion due to facial paralysis. Ten cases showed a favorable outcome following surgery, with stable results seen over an average follow-up period of 4.5 years. In one case, recurrence of ectropion was observed 2 months after surgery due to an insufficient correction, and the patient required repeat surgery. Conclusions The orbicularis oculi muscle flap was an effective means of suspension and was able to maintain long-term traction tension. This procedure can therefore be considered a favorable treatment option for lower eyelid ectropion due to facial paralysis.

Developmental lesions, including benign or malignant tumors, rarely arise in the masseteric region. This retrospective study was performed to evaluate the adequacy of different surgical approaches in a series of patients with lesions in the masseteric region. The surgical methods, postoperative complications, pathological diagnosis, and aesthetic outcome were compared in 4 patients who underwent excision of intramasseteric mass lesions. A flap was elevated in 3 patients and direct incisional resection was performed in 1 patient. Two patients underwent incisional biopsy to exclude malignancy before the total excision was performed. The final diagnosis was hemangioma in 3 patients and schwannoma in 1 patient. All patients were satisfied with the aesthetic results and there were no major complications. In conclusion, surgery with skin flap elevation was concluded to be the best approach based on overall assessment of technical considerations, complications, and the cosmetic outcome. However, the surgeon should not hesitate to perform direct incisional biopsy when malignancy needs to be excluded.

Although human recombinant basic fibroblast growth factor (bFGF) is widely used for wound healing, daily treatment with bFGF is required because of its short half-life. An effective controlled-release system of bFGF is, therefore, desired in clinical settings. To investigate the efficacy of a bFGF-loaded nanosheet for wound healing, focusing on the controlled-release of bFGF, bFGF-loaded poly(lactic-co-glycolic acid) (PGLA) nanosheets were developed, and their in vitro release profile of bFGF and their in vivo efficacy for wound healing were examined. A polyion complex of positively charged human recombinant bFGF and negatively charged alginate was sandwiched between PLGA nanosheets (70 nm thick for each layer). The resulting bFGF-loaded nanosheet robustly adhered to silicon skin by observation using a microscratch test. bFGF was gradually and continuously released over three days in an in vitro incubation study. Treatment with the bFGF-loaded nanosheets (every 3 day for 15 days) as well as with a conventional bFGF spray effectively promoted wound healing of mouse dorsal skin defects with accelerated tissue granulation and angiogenesis, although the dose of bFGF used in the treatment with the bFGF nanosheets was approximately 1/20 of the sprayed bFGF. In conclusion, we developed a bFGF-loaded nanosheet that sustained a continuous release of bFGF over three days and effectively promoted wound healing in mice.

Background Microvascular anastomosis using cyanoacrylate adhesive has a reputation among researchers as an alternative to conventional sutures. However, a degree of ingenuity is required to avoid the collapse of the vascular lumen for the duration of the anastomosis. The aim of this study was to determine the feasibility of intravascular stenting (IVaS) as a temporary stent during sutureless microvascular anastomosis with cyanoacrylate adhesive. Methods Sixty male Fisher 344 rats were evenly divided into two groups. The right superficial femoral arteries (RSFAs) were transected in each group. Microvascular anastomoses were then performed with the sutureless (SL) method in one group and conventional sutures (CS) in the other group. The diameter of the RSFA, duration of microvascular anastomosis, and the patency of the RSFA were evaluated immediately after anastomosis and 7 days after the surgery. Tissue samples were obtained for pathological consideration. Results There was no significant difference in the diameter of the RSFAs between the SL and the CS groups. There was no significant difference in the patency rates of the groups. The anastomosis time of the SL group was significantly shorter than that of the CS group, regardless of the experience of the surgeons. A histological analysis showed a comparable level of foreign body reactions in each group. Conclusion IVaS plays a supportive role in sutureless microvascular anastomosis with cyanoacrylate adhesive. The short-term safety of this technique has now been confirmed at the experimental stage.

Hypochlorous acid (HOCl) solution (200 ppm, pH 6) was prepared and evaluated for their stabilities and microbicidal activities. We demonstrated that HOCl is unstable against ultraviolet (UV) light, sunshine, contact with air, and elevated temperature (≧25℃). Furthermore, in the HOCl solution, the presence of excess NH2- or CHO-containing organic compounds such as proteins and carbohydrates, or of inorganic ions such as NO2-, SO3-, PO3-, Fe2+, Cu2+, and CuS, resulted in the rapid consumption of HOCl by oxidation reactions, and significantly decreased the microbicidal activity of the HOCl solution against coliform bacteria and total viable cell count. Thus, production of stable HOCl solution requires formulation in pure water harboring concentrations as low as possible of various compounds and ions, as well as storage in dark and cool conditions (

Background Frey syndrome is one of the important complications of parotid surgery. Placing a barrier between the parotid gland and the skin flap is considered to be effective for preventing this syndrome by blocking misdirected regeneration of nerve fibers. Neoveil is nonwoven polyglycolic acid (PGA) felt that is used as absorbable reinforcing material. The present study evaluated the effectiveness of this PGA felt as a barrier for preventing Frey syndrome. Methods A total of 19 patients who underwent superficial parotidectomy from 2012 to 2017 were retrospectively reviewed. Application of PGA felt (n = 9) or sternocleidomastoid flap (SCMF) transfer (n = 10) was performed to prevent Frey syndrome. The total operating time, intraoperative blood loss, operating time after parotidectomy, and postoperative drainage volume were compared between the 2 groups. Results The operating time after parotidectomy was significantly shorter in the PGA felt group than in the SCMF group (43.7 ± 10.7 vs 57.8 ± 15.7 minutes). The total operative time was shorter and the drainage volume was smaller in the PGA felt group than those in the SCMF group without statistical significance (210.1 ± 43.4 vs 234.3 ± 52.7 minutes and 80.6 ± 36.8 vs 118.8 ± 83.9 mL). Neither Frey syndrome nor other major complications occurred in either group. Conclusions Use of PGA felt successfully prevented Frey syndrome, while being technically simpler and more rapid compared with SCMF transfer.

When frostbite thaws, reperfusion injury has a crucial impact on tissue injury, and production of free radicals induces further tissue damage. This study examined whether extract of Ginkgo biloba 761 could ameliorate frostbite injury as a free radical scavenger.Seventy-five Fisher 344 rats were divided into five groups of 15, and frostbite injury was created in each animal by sandwiching the left hind foot between a frozen magnet (-78.5°C) and a room-temperature magnet. Group I received saline; groups II, III, and IV received extract of Ginkgo biloba 761 (200, 100, and 50 mg/kg, respectively); and group V received superoxide dismutase (12 mg/kg). All drugs were injected intraperitoneally three times at 24-hour intervals. The wound surface area was measured throughout the wound healing period. Wounds were also harvested at various times to count cells stained by monoclonal antibodies for 4-hydroxy-2-nonenal and 8-hydroxy-2'-deoxyguanosine.Compared to group I, the wound surface area was significantly smaller in groups II and III on days 1 and 3 after wound creation. Histologic examination revealed significantly more 4-hydroxy-2-nonenal-stained cells and 8-hydroxy-2'-deoxyguanosine-stained cells in group I compared to other groups on day 1. However, there was no difference in the total healing period among the groups. A higher dose test of extract of Ginkgo biloba 761 (300 mg/kg daily) induced animal death, probably because of toxicity.Extract of Ginkgo biloba 761 demonstrated a protective effect against frostbite in the present model and probably alleviated reperfusion injury by reducing tissue peroxidation.

Weakly acidic hypochlorous acid (HClO; 200 ppm, pH 6.5) is effective against a broad range of microorganisms. We have previously reported a study of developing antimicrobial biomaterials made up of chitin-nanofiber sheet (CNFS) -immobilized silver nanoparticles (CNFS/Ag NPs) and showed that either cleansing with HClO or covering with CNFS/Ag NPs daily for more than 7 days resulted in delayed wound healing. This study aimed to evaluate disinfection and wound healing by a combination of cleansing with HClO and covering with CNFS/Ag NPs daily for 3 days. Applying HClO + CNFS/Ag NPs daily for 3 days and then cleansing with just pure water and covering with CNFS alone daily for 9 days were performed for Pseudomonas aeruginosa-infected wounds in db/db diabetic mice. We found a significant enhancement of wound healing and a reduction of bacteria counts compared to the controls. Histological examination showed significantly advanced granulation tissue and capillary formations in the wounds on Day 12. These results suggest that limited disinfection to 3 days with HClO + CNFS/Ag NPs may be sufficient to avoid negative effects on wound repair.

UVB exposure penetrates deeply into the dermis to alter skin barrier function, which is a primary factor in skin photoaging. We previously reported that dalteparin and protamine nanoparticles (D/P NPs) are effective carriers of FGF-2. This study aimed to examine the ability of FGF-2-containing D/P NPs (FGF-2D/P NPs) to ameliorate UVB-induced skin photoaging in hairless mice. Dorsal skin of HR-1 hairless mice were exposed to UVB irradiation 5 days/week for 8 weeks (UV (+): final total, 2700 mJ/cm2). Mice were divided into four groups: Non-UVB (UV (-)) + saline, UV (+) + saline, UV (+) + FGF-2D/P NPs, UV (+) + FGF-2, and UV (+) + D/P NPs, and following UVB irradiation, FGF-2D/P NPs, FGF-2, and D/P NPs were applied to the groups of mice just after each UVB irradiation. Each group was subjected to evaluation of skin changes (elasticity), and histological examination using hematoxylineosin and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. UVB irradiation of mice significantly induced a decline in elasticity and acanthosis, which was alleviated by application of FGF-2D/P NPs. Furthermore, TUNEL-staining showed the proportions of apoptotic dermal fibroblast cells (DFCs) and epidermal keratinocyte cells (EKCs) in the UV (+) + FGF-2D/P NPs group were significantly lower than those in the UV (+) + saline, UV (+) + FGF-2, and UV (+) + D/P NPs groups. Thus, FGF-2D/P NPs may be effective in preventing skin photoaging accelerated by UVB irradiation such as declining elasticity, acanthosis, and apoptosis of DFCs and EKCs.

Application of a high-velocity steam-air micromist jet spray (HVS-AMJS; micromist average diameter: 2.4 μm) for cleansing the skin is proposed. Low-pressure steam is mixed with compressed air (pH 6.5) in a nozzle, and then sprayed at a pressure of ≦0.25 MPa and a velocity of ≧0.34 m/s on the skin or surface of material located approximately 5-10 cm from the nozzle. The temperature on the sprayed surface and water flow rate could be controlled between 42 °C and 46 °C and at approximately 50 mL/min, respectively. Compared with ultrasonic cleansing with tap water and rubbing with only tap water, the HVS-AMJS successfully removed fluorescent lotion covering pieces of wood and significantly reduced both the number of coliforms and the total viable counts on pieces of wood and gauze. Furthermore, the HVS-AMJS effectively removed oily ink from the skin of hairless rats, and temporarily elevated the skin temperature and blood flow, indicating massage effects. The striking characteristics of this cleansing technique using HVS-AMJS are not only its ability to remove microbes and residue without using any chemicals or detergents but also its massage effects.

Although human recombinant basic fibroblast growth factor (bFGF) is widely used for wound healing, daily treatment with bFGF is required because of its short half-life. An effective controlled-release system of bFGF is, therefore, desired in clinical settings. To investigate the efficacy of a bFGF-loaded nanosheet for wound healing, focusing on the controlled-release of bFGF, bFGF-loaded poly(lactic-co-glycolic acid) (PGLA) nanosheets were developed, and their in vitro release profile of bFGF and their in vivo efficacy for wound healing were examined. A polyion complex of positively charged human recombinant bFGF and negatively charged alginate was sandwiched between PLGA nanosheets (70 nm thick for each layer). The resulting bFGF-loaded nanosheet robustly adhered to silicon skin by observation using a microscratch test. bFGF was gradually and continuously released over three days in an in vitro incubation study. Treatment with the bFGF-loaded nanosheets (every 3 day for 15 days) as well as with a conventional bFGF spray effectively promoted wound healing of mouse dorsal skin defects with accelerated tissue granulation and angiogenesis, although the dose of bFGF used in the treatment with the bFGF nanosheets was approximately 1/20 of the sprayed bFGF. In conclusion, we developed a bFGF-loaded nanosheet that sustained a continuous release of bFGF over three days and effectively promoted wound healing in mice.

Background Activated platelet-rich plasma secrets many growth factors (GFs), and low-molecular weight heparin-protamine micro/nanoparticles (LMWH-P M/NPs) significantly interact with, enhance, and stabilize the secreted GFs. Objective The purpose of this study was to evaluate the effects of LMWH-P M/NPs and GFs (from platelet-rich plasma) on full-thickness skin graft (FTSG). Methods A total of 96 inbred male rats were anesthetized and 4-cm full-thickness skin wound were created on dorsal skin of rats. LMWH-P M/NPs and GFs, LMWH-P M/NPs, GFs and saline (control) were then injected evenly into cutaneous muscles at the wound. The next day, the rats underwent FTSG. On the indicated days after FTSG, blood flow of FTSG site (wound bed and FTSG) was examined by 2-dimensional laser Doppler blood flowmeter. On 10 days, pictures of FTSG site were taken and FTSG survival rate was evaluated. Histologic analyses of skin samples were performed on 4, 7, and 10 days. Results Treatment of full-thickness skin wound with LMWH-P M/NPs and GFs effectively promoted survival rate of FTSG and blood flow of FTSG site compared with those treated with GFs, LMWH-P M/NPs, and control. LMWH-P M/NPs and GFs also promoted new vessel formation at FTSG site. Conclusions The prior injection of LMWH-P M/NPs and GFs into wound bed increases FTSG survival rate, and promotes blood flow and angiogenesis at FTSG site.

Fragmin/protamine microparticles (F/P MPs) have been used as carriers for the preservation and activation of cytokines in human plasma (HP)–Dulbecco’s modified Eagle’s medium (DMEM) gels. This study investigated a three-dimensional (3D) culture system using an HP–DMEM gel with 0.1 mg/mL F/P MPs and 5 ng/mL FGF-2 for the proliferation of human dermal fibroblast cells (DFCs), human microvascular endothelial cells (MVECs) and human coronary smooth muscle cells (SMCs), or 5 ng/mL interleukin (IL)-3/granulocyte-macrophage colony-stimulating factor (GM-CSF) for a human hematopoietic cell line (TF-1 cells). DFCs, MVECs, SMCs and TF-1 cells grew rapidly under 3D culture conditions using a low-concentration HP (2 %)–DMEM gel with F/P MPs and FGF-2 (for DFCs, MVECs and SMCs) or IL-3/GM-CSF (for TF-1 cells) at doubling times of 22, 23, 25 and 18 h, respectively, without the use of animal serum, compared to under 2D culture conditions using low-concentration human serum (2 %)–DMEM with 5 ng/mL FGF-2 or IL-3/GM-CSF on F/P MP-coated plates at doubling times of approximately 26, 25, 40 and 20 h, respectively.

Conventional surgery of toe syndactyly is accomplished using a zig-zag incision and full thickness or split thickness skin grafting. However, this method has an aesthetic problem with a prominent scar on the dorsal side of the foot. The aim of the article was to investigate the efficacy of a cosmetically conscious operation for syndactylies performed using a single technique.A retrospective analysis was performed involving 66 patients with 88 syndactylic toes who presented to the department of plastic and reconstructive surgery at 1 of 3 major institutions during a 16-year period (1998-2014). Syndactylies were classified according to the affected interdigital spaces (IDSs). All cases were treated using the same method: linear skin incisions to divide the digits, triangular flaps on the dorsal and plantar sides of the foot, and preserved subcutaneous vascular network skin grafts from the medial inframalleolar region.The most frequent type of toe syndactyly was IDS-2 (union of the second/third toes), which accounted for 59.1% (39/66) of the cases. The rates of familial anomalies were significantly different between bilateral IDS-2 (4/13) and unilateral IDS-2 (0/24) (P0.05, Fisher exact test). There were no severe postoperative complications, including scar contracture or keloid formation. All patients obtained sufficiently deep IDSs, and none required repeat surgery.A straight incision minimizes the scar observed from the visible side, and preserved subcutaneous vascular network skin grafts prevent postoperative contracture of linear scar.

Yuki Takabayashi,1Masaki Nambu,1Masayuki Ishihara,2Masahiro Kuwabara,1Koichi Fukuda,2Shingo Nakamura,2Hidemi Hattori,2Tomoharu Kiyosawa1 1Department of Plastic and Reconstructive Surgery, 2Division of Biomedical Engineering, Research Institute, National Defense Medical College, Tokorozawa, Saitama, Japan Purpose: Although treatments for alopecia are in high demand, not all treatments are safe and reliable. Dalteparin/protamine nanoparticles (D/P NPs) can effectively carry growth factors (GFs) such as fibroblast GF (FGF)-2. The purpose of this study was to identify the effects of FGF-2-containing D/P NPs (FGF-2&D/P NPs) on hair growth.Patients and methods: In this study, the participants were 12 volunteers with thin hair. One milliliter of FGF-2 (100 ng/mL) and D/P NPs (56 μg/mL) was applied and massaged on the skin of the scalp by the participants twice a day. They were evaluated for 6months. Participants were photographed using a digital camera for general observation and a hair diagnosis system for measuring hair diameter.Results: The mean diameter of the hairs was significantly higher following the application of FGF-2&D/P NPs for 6months. Objective improvements in thin hair were observed in two cases. Nine participants experienced greater bounce and hair resilience.Conclusion: The transdermal application of FGF-2&D/P NPs to the scalp can be used as a new treatment for alopecia. Keywords: hair growth, dalteparin/protamine nanoparticles, fibroblast growth factor, transdermal application

Occlusive dressing is widely accepted and used to manage skin ulcers. However, with respect to its application to incisional wounds, most studies have been conducted about the clinical effects on incisional healing of surgical sites. Studies of the histological effects of occlusive dressing for incisional wounds have been few. The aim of this study was to clarify the histological effects of occlusive dressings on healing of incisional skin wounds. Rat dorsal skin was incised down to the panniculus and sutured immediately. Dressing types included 2-octyl cyanoacrylate and hydrocolloid materials as occlusive dressings and no-dressing as the open therapy. Histological examination and dermoscopic observation were performed 1, 2, 4 and 7 days after surgery. The findings from each dressing type were compared. In the open therapy group, the upper portion of the edge of incision was necrosed minimally and finally healed with wide scar formation. However, in the occlusive dressing groups, micronecrosis of the incision edge seen in the no-dressing group was not observed, healing was more rapid and the remaining scar was finer. Occlusive dressing can prevent micronecrosis of the incision edge, resulting in rapid and excellent healing. This study shows that the efficacy of and supports the use of occlusive dressing in incisional wound management.

The dry weight of tendon tissue is accounted for mainly by collagen fibers. Accordingly, the tendon-healing process primarily involves repair of collagen fibers. During the remodeling phase of tendon healing, newly proliferating collagen fibers are transformed into a mature repaired tendon. Despite the importance of this phenomenon, the details of fibrous rebuilding have not been reported previously. The aim of this study was to visualize the ultrastructural changes and to obtain a clear understanding of the reorganization of the collagen fibers in the tendon repair site, using rat Achilles tendons. We used scanning electron microscopy (SEM) with cell maceration as the main method of analysis. Pretreatment with cell maceration removed the cellular components successfully. This allowed precise visualization of each collagen fiber and the three-dimensional network of the fibers. This study was the first to apply the cell-maceration/SEM method to observe tendon tissue. Seven days after surgery, new collagen fibers grew extensively in the repair site in a random arrangement. Fourteen days after surgery, the collagen fibers began to form an axial arrangement. Near the tendon stump, this change progressed from the outer layer to the core region. On the other hand, in the middle of the repair site, it progressed from the core to the outer layer. Change in the axial arrangement of collagen fibers contributes to the connection between the repair site and the tendon stump and to the separation of the repair site from the paratenon.

We examined the effects of prostaglandin E₁ (PGE₁) on radiation-induced proliferation inhibition and apoptosis in keratinocytes and healing of radiation-induced skin injury in a rat model. PGE₁ had a protective effect on radiation-induced growth inhibition in keratinocytes in vitro, but not in fibroblasts. Varying concentrations of PGE₁ were subcutaneously administered into the posterior neck region. X-irradiation at a dose of 20 Gy was administrated to the lower part of the back using a lead sheet with two holes 30 min to 1 h before or after the administration of PGE₁. Although X-irradiation induced epilation, minor erosions, or skin ulcers in almost all rats, PGE₁ administration prior to irradiation reduced these irradiation injuries. Staining with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling showed that proportions of apoptotic keratinocytes in the X-irradiated skin of PGE₁-administered rats were significantly lower than for those in the skin of rats which did not receive PGE₁. Cutaneous full-thickness defective wounds were then formed in X-irradiated areas to examine the time course of wound healing. Wound healing was significantly delayed because of X-irradiation, but PGE₁ administration prior to irradiation led to a significantly shorter delay in wound healing compared with controls. Decreasing delay in wound healing was correlated with concentration of PGE₁ administrated. Thus, PGE₁-administration may potentially alleviate the radiation-induced skin injury.

Fragmin/protamine nanoparticles (F/P NPs) have been used as carriers for the preservation and controlled release of fibroblast growth factor (FGF)-2 and various cytokines in human plasma (HP). This study tested an HP–Dulbecco's modified Eagle's medium (DMEM) gel as a three-dimensional (3D) culture for the expansion of adipose tissue-derived multilineage stromal cells (ASCs) and bone marrow-derived mesenchymal stem cells (BMSCs). The growth of these cells improved in 3D culture using low-concentration HP (2%)–DMEM gel with 0.1 mg/mL F/P NPs and 5 ng/mL FGF-2 without animal serum in comparison to two-dimensional (2D) culture using a low-concentration human serum (2%)–DMEM containing 5 ng/mL FGF-2 on F/P NPs-coated plates. ASCs and BMSCs, which were expanded in the low-concentration HP–DMEM gel with F/P NPs and FGF-2, maintained their multilineage potential for differentiation into adipocytes or osteoblasts similar to the 2D cultured cells. Furthermore, flow cytometric analyses showed that the phenotypic markers which were positive for CD44, CD90, and CD105 (>80%) and negative for CD34 and CD45 (

Background Skin flap necrosis is a problem encountered postoperatively. The purpose of this study was to evaluate the effects of platelet-rich plasma containing fragmin/protamine microparticles (PRP&F/P MPs) on viability in a rat dorsal paired pedicle skin (DPPS) flap. Materials and Methods Two symmetrical adjoining rectangular flaps (8 × 2 cm each) were drawn on the rat dorsum. Two days after PRP&F/P MPs-, PRP-, F/P MPs-, and saline (control)-injections ( n = 8 each), flaps were elevated as a random pattern flap without the lateral thoracic, posterior intercostal, and deep circumflex iliac vessels. The flaps were immediately sutured back and the flap survival area was measured 7 d after flap elevation. Results The flap survival rate in PRP&F/P MPs-injected groups (73.1% ± 4.2%) was significantly higher than those in PRP (64.9% ± 4.0%), F/P MPs (59.4 ± 4.5%), and control (61.2% ± 4.2%) groups. Histologic observation of the flaps showed survived thick granulation tissue and neovascularization in PRP&F/P MPs-injected groups. Conclusions When PRP&F/P MPs are administered 2 d before the flap elevation, the improved flap survivals are observed. The pre-injection of PRP&F/P MPs may thus represent a promising treatment to prevent skin flap necrosis in reconstructive surgery.

The purpose of this study was to evaluate effects of human platelet-rich plasma (PRP)-containing fragmin/protamine microparticles (F/P MPs) as a protein carrier on neovascularization and granulation tissue formation. Frozen and thawed PRP contains high concentrations of various growth factors (GFs) and F/P MPs effectively adsorb those GFs. Human microvascular endothelial cells (MVECs) and dermal fibroblast cells (DFCs) were optimally grown in medium containing 4% PRP and the addition of F/P MPs significantly maintained and protected the proliferative activity of PRP incubated at 37°C for more than 10 days. When PRP-containing F/P MPs were subcutaneously injected into the back of mice, significant neovascularization was induced near the injected site with enhanced filtration of inflammatory cells from day 3 to day 30, compared with controls (injections of PRP, F/P MPs, and saline). Both PRP-containing F/P MPs and PRP alone induced significant formation of granulation tissue at the injected site. However, thickness of induced granulation tissues was well maintained for 30 days only in PRP-containing F/P MP-injected group. Those bound GFs may be gradually diffused and released from F/P MPs in vitro and in vivo. Thereby, PRP-containing F/P MPs offer significantly higher inductions of vascularization and fibrous tissue formation in vivo than PRP alone.

We evaluated the effects of fragmin/protamine micro-particles (F/P MPs) containing FGF-2 (F/P MP-F) as carriers for the controlled release of FGF-2 for adipocyte-survival and capillary formation in inbred rats with subdivided free fat grafts. F/P MPs could immobilize FGF-2, thereafter gradually releasing the bound FGF-2. Inbred Fisher 344 rats weighing around 150 g were anesthetized and implanted with paste comprising harvested fat combined with F/P MP-F. The effect of F/P MP-F on the survival, granulation, and capillary formation in fat grafts was histologically compared with control grafts containing either FGF-2, F/P MPs or PBS. The control fat grafts became attached to tissues adjacent to the implantation site and were significantly resorbed after 30 days. In contrast, pink, soft, supple grafts were compressible and were little resorbed in the group given F/P FP MP-F at 30-120 days. Normal adipocytes were obviously decreased in the control groups with increased granulation tissues, whereas normal adipocytes with capillary formations were maintained in the F/P MP-F group. Thus, adding F/P MP-F to subdivided fat grafts helps to improve graft volume retention and survival in soft-tissue reconstruction through accelerating adipocyte-survival rates and angiogenesis.

To create a moist environment for rapid wound healing, a hydrosheet composed of alginate, chitin/chitosan, and fucoidan (ACF-HS) has been developed as a functional wound dressing. The aim of this study was to evaluate the accelerating effect of ACF-HS on wound healing for rat mitomycin C-treated healing-impaired wounds. Full-thickness skin defects were made on the back of rats and mitomycin C was applied onto the wound for 10 minutes to prepare a healing-impaired wound. After thoroughly washing out the mitomycin C, ACF-HS was applied to the healing-impaired wounds. The rats were later euthanized and histological sections of the wounds were prepared. The histological examinations showed significantly advanced granulation tissue and capillary formations in the healing-impaired wounds treated with ACF-HS on days 7 and 14, in comparison with that in alginate fiber (Kaltostat), hydrogel wound dressing (DuoACTIVE), and nontreatment (negative control). Furthermore, in cell culture studies, ACF-HS-absorbed serum and fibroblast growth factor-2 was found to be proliferative for fibroblasts and endothelial cells, respectively, and ACF-HS-absorbed serum was found to be chemoattractive for fibroblasts. However, our results may not be strictly comparable with general healing-impaired wound models in humans because of the cell damage by mitomycin C. In addition, more biocompatibility studies of fucoidan are essential due to the possibility of renal toxicity.