Long-Term Follow-up With Multispecialty Management of a Giant Lymphangioma of an Infant Tongue Contributed to Reduced Complications of the Disease: A Case Report of a 21-Year Follow-up.

Background. Lymphangiomas are benign tumors of abnormal lymphatic tissue. Approximately 6% of all lymphangiomas occur on the tongue. A lymphangioma of the tongue may present as a localized or a diffused growth, which may enlarge to cause macroglossia, impaired speech, and difficulty in mastication. This article reports a 21-year follow-up of a male infant who presented with a giant tongue lymphangioma. This long-term follow-up with multidisciplinary management including partial glossectomy, sclerotherapy, and orthodontic treatment to diminish complications of the disease in adulthood.

The anatomic classification of the anophthalmic eye socket (Types 0-V): A high-level taxonomy classification system.

Eyelid and orbital deformities caused by various defects, such as tumor resection and trauma, whether congenital or acquired, are extremely complex and atypical. Several classifications systems for these anophthalmic conditions have been developed, but none have proven suitable for determining the ideal reconstructive surgical strategy. We therefore designed a novel system known as the Anatomic Classification of the Anophthalmic Eye Socket (Types 0-V) for optimum reconstruction of anophthalmic socket deformities focusing on eyelid and orbital defects. Type 0 is an eyelid defect type, in which only the eyelid is defective; Type I is a phthisis bulbi type, in which the anatomical eyeball is preserved, but the visual function is lost; Type II is a substitution type involving filling with implant packing after evisceration or enucleation; Type III is an enucleation type involving enucleation of the ophthalmus; Type IV is an exenteration (incomplete) type involving exenteration while saving the eyelids; and Type V is an exenteration (complete) type involving exenteration including eyelid defects. Using this classification system, it will be possible to standardize reconstructive procedures of the anophthalmic socket.

Auricular Cartilage Composite Graft for Glans Reconstruction After Squamous Cell Carcinoma of the Penis.

Background. Surgical procedures for squamous cell carcinoma of the penis generally involve primary closure, partial glansectomy, skin graft, and penile amputation. Partial penile resection can result in not only unsightly deformation of the penis but also functional disorders of the urinary line as well as psychological effects due to subjective perceptions of a loss of power and masculinity. With the use of an organ-preserving procedure for functional reconstruction without compromising oncological control, this report describes a new procedure for performing functional penile reconstruction with an auricular cartilage composite graft.

Functional Reconstruction for Severe Heat Press Injury of the Proper Digits Using Immediate Preserved Subdermal Vascular Network Skin Graft: A Case Report.

The patient was a 57-year-old woman who suffered third-degree burns to the dorsal side of her left fingers (non-dominant hand) when it was caught in a press machine heated to 200°C while she was cleaning clothes. At 20 hours after the injury, the necrotic tissue was excised and covered with a preserved subdermal vascular network (PSVN) skin graft, which allowed the patient to regain a very good hand function. Even for severe heat press injuries of the hand, the combination of immediate surgical intervention, a thick skin graft with PSVN, and early rehabilitation from the first postoperative week leads to good functional and cosmetic reconstruction outcomes.

Giant sebaceous nevus occupying half of the head associated with sebaceous carcinoma, apocrine adenocarcinoma, and basal cell carcinoma: A case report.

We herein report a 57-year-old man afflicted with multiple malignant tumors arising from a giant sebaceous nevus on his left parieto-temporal scalp and neck. The upper eyelid was resected in all layers, and the auricle was resected as well, leaving part of the external auricular cartilage; the parotid gland area was also resected over the parotid fascia. The lost part of the left eyebrow was reconstructed using an anterior forehead skin flap, and the residual defect was covered with skin graft and expanded scalp flap. This is probably the first case report of giant sebaceous nevus associated with three malignancies: sebaceous carcinoma, apocrine adenocarcinoma, and basal cell carcinoma.

A novel technique for forehead and eyebrow reconstruction using a temporal hairline flap combined with a free forearm flap: a report of two cases.

We herein report a novel technique for managing large forehead skin defects that include the eyebrows and upper eyelid. Case 1 was of a 55-year-old woman with an eccrine carcinoma (T2N0M0) at the right forehead, resulting in a skin defect 15 cm in diameter. Case 2 was of an 81-year-old man with a malignant peripheral nerve sheath tumor (T1a N0M0) at the left forehead, resulting in a skin defect 18 cm in diameter. A pedicled temporal hairline flap and free forearm flap were combined and placed on the defect, and good eyebrows and forehead morphology was ultimately achieved.

Rare cases requiring emergency procedures while harvesting a free forearm flap: report of two cases.

The free forearm flap is considered safe to harvest and is still extremely useful for reconstruction after head and neck tumor resection. We experienced two uncommon cases, in which an emergency procedure was required during reconstructive surgery after resection of head and neck cancer. Case 1 suffered persistent hand blood flow insufficiency long after the flap was placed in the oral cavity. Case 2 had an anomalous cutaneous vein of the forearm that was a drainage vein of the flap. For risk management during free flap surgery, preparing multiple venous transplant options in advance is fundamental.

Cultured epithelial autografts for the treatment of large-to-giant congenital melanocytic nevus in 31 patients.

INTRODUCTION: Giant congenital melanocytic nevus (GCMN) is a large melanocytic nevus, and its full-thickness removal is usually difficult due to the lack of skin available for reconstruction. Curettage is an alternative approach in cases of GCMN to remove the superficial dermis above the cleavage plane with a curette in the neonatal period, and its major complications include repigmentation, retarded epithelization, and hypertrophic scar formation. In Japan, the JACE® cultured epidermal autograft (CEA) was approved and covered by public healthcare insurance for the treatment of congenital melanocytic nevus (CMN) that is difficult to treat with conventional methods in 2016. We have used CEA for wounds after curettage in the neonatal period or following ablation after the neonatal period in combination with laser therapies to reduce the above-mentioned complications. METHODS: In this study, we summarized all consecutive CMN patients treated using CEA from December 2016 to April 2019 and evaluated the duration required for epithelialization, incidence of hypertrophic scar, and color change in the target nevus by comparing the L∗ values one year later between the Curettage group, the non-Curettage group with initial treatment or the subsequent group. RESULTS: No significant differences were seen in the epithelization period or incidence of hypertrophic scars among the groups, but the color of the target nevus was improved significantly in the Curettage group (p < 0.01) and non-Curettage group with initial treatment (p < 0.01). CONCLUSIONS: In conclusion, CEA seems to accelerate epithelization after curettage or ablation of CMN, and this treatment could improve the color of CMN when applied initially.

Urethral functional reconstruction using a pedicled thigh flap for urethral cutaneous fistula after vulvar necrotizing fasciitis.

The penile and scrotal skin are often resected after vulvar necrotizing fasciitis, so functional and aesthetic vulvar reconstruction is extremely challenging. With widespread and deepened infections, urethral fistulas can develop, forcing penis and scrotal amputations. In cases complicated with a urethral fistula, the wound cannot be treated by simple skin graft surgery, and complicated flap surgery is required. We verified that urethral functional reconstruction using a pedicled thigh flap is a useful technique for managing urethral cutaneous fistulas that develop after necrotizing fasciitis.

A Novel Treatment for Giant Congenital Melanocytic Nevi Combining Inactivated Autologous Nevus Tissue by High Hydrostatic Pressure and a Cultured Epidermal Autograft: First-in-Human, Open, Prospective Clinical Trial.

BACKGROUND: Giant congenital melanocytic nevi are large skin lesions associated with a risk of malignant transformation. The authors developed a novel treatment to reconstruct full-thickness skin defects by combining an inactivated nevus as the autologous dermis and a cultured epidermal autograft. The first-in-human trial of this treatment was performed. METHODS: Patients with melanocytic nevi that were not expected to be closed by primary closure were recruited. The full-thickness nevus of the target was removed and inactivated by high hydrostatic pressurization at 200 MPa for 10 minutes. The inactivated nevus was sutured to the original site, and a cultured epidermal autograft was grafted onto it 4 weeks later. Patients were followed for up to 52 weeks. RESULTS: Ten patients underwent reimplantation of the pressurized nevus, and one patient dropped out. The recurrence of nevus at 52 weeks was not detected by pathological diagnosis in any patients. The L* value at 52 weeks was significantly higher than that of the target nevus. One patient received skin grafting due to contracture of the reconstructed skin. The epithelized area of the reconstructed skin, as the percentage of the original target nevus, was 55.5 ± 19.4 percent at 12 weeks and 85.0 ± 32.4 percent at 52 weeks. CONCLUSIONS: The inactivated nevus caused inflammation and contracture for several months. However, no recurrence was observed, and combination therapy using an inactivated nevus with a cultured epidermal autograft may therefore be a novel treatment of giant congenital melanocytic nevi. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.

Reconstruction of whistling deformity using V-Y advancement flap after primary cleft lip repair.

Whistling deformity often occurs after cleft lip primary repair, marked by deficiency of vermilion tubercle. The purpose of the surgery includes improving the appearance and function of the lip. Adequate amount of vermillion tubercle is necessary to fully cover the front teeth, produce lip seal while speaking consonant sound and balance the projection of face. Many techniques are known to repair whistling deformity from fat injection to Abbe flap. In our department, we use V-Y advancement flap. At certain cases, we repeat the procedure to maximize the result. This simple and less complication technique shows stable outcome during follow up. We recommend this technique because of the non-visible scar after surgery, applicable for unilateral and bilateral cleft lip case at various age, and possible to be done under general or local anesthesia.

GALA-Modified Lipid Nanoparticles for the Targeted Delivery of Plasmid DNA to the Lungs.

This study describes the development of lipid nanoparticles (LNPs) for the efficient and selective delivery of plasmid DNA (pDNA) to the lungs. The GALA peptide was used as a ligand to target the lung endothelium and as an endosomal escape device. Transfection activity in the lungs was significantly improved when pDNA was encapsulated in double-coated LNPs. The inner coat was composed of dioleoylphsophoethanolamine and a stearylated octaarginine (STR-R8) peptide, while the outer coat was largely a cationic lipid, di-octadecenyl-trimethylammonium propane, mixed with YSK05, a pH-sensitive lipid, and cholesterol. Optimized amounts of YSK05 and GALA were used to achieve an efficient and lung-selective system. The optimized system produced a high gene expression level in the lungs (>107 RLU/mg protein) with high lung/liver and lung/spleen ratios. GALA/R8 modification and the double-coating design were indispensable for efficient gene expression in the lungs. Despite the fact that NPs prepared with 1-step or 2-step coating have the same lipid amount and composition and the same pDNA dose, the transfection activity was dramatically higher in the lungs in the case of 2-step coating. Surprisingly, 1-step or 2-step coatings had no effect on the amount of nanoparticles that were delivered to the lungs, suggesting that the double-coating strategy substantially improved the efficiency of gene expression at the intracellular level.

Ex vivo Induction of Apoptotic Mesenchymal Stem Cell by High Hydrostatic Pressure.

Among promising solutions for tissue repair and wound healing, mesenchymal stem (or stromal) cells (MSCs) have been a focus of attention and have become the most clinically studied experimental cell therapy. Recent studies reported the importance of apoptosis in MSC-mediated immunomodulation, in which apoptotic MSCs (apoMSCs) were shown to be superior to living MSCs. Nowadays, high hydrostatic pressure (HHP), a physical technique that uses only fluid pressure, has been developed and applied in various bioscience fields, including biotechnology, biomaterials, and regenerative medicine, as its safe and simply operation. In the current study, we investigated the impact of HHP treatment on human bone marrow-MSC survival and proliferation. Based on the detection of executioner caspase activation, phosphatidylserine exposure, DNA fragmentation (TUNEL) and irrefutable ultrastructural morphological changes on transmission electron microscopy (TEM), our data revealed that HHP treatment induced complete apoptosis in MSCs. Notably, this technique might provide manipulated products for use in cell-based therapies as manufacturing capability expands. We hope that our findings will contribute to the improvement of MSCs or EVs in translational research development. Graphical Abstract.

Hydrostatic pressure can induce apoptosis of the skin.

We previously showed that high hydrostatic pressure (HHP) treatment at 200 MPa for 10 min induced complete cell death in skin and skin tumors via necrosis. We used this technique to treat a giant congenital melanocytic nevus and reused the inactivated nevus tissue as a dermis autograft. However, skin inactivated by HHP promoted inflammation in a preclinical study using a porcine model. Therefore, in the present study, we explored the pressurization conditions that induce apoptosis of the skin, as apoptotic cells are not believed to promote inflammation, so the engraftment of inactivated skin should be improved. Using a human dermal fibroblast cell line in suspension culture, we found that HHP at 50 MPa for ≥ 36 h completely induced fibroblast cell death via apoptosis based on the morphological changes in transmission electron microscopy, reactive oxygen species elevation, caspase activation and phosphatidylserine membrane translocation. Furthermore, immunohistochemistry with terminal deoxynucleotidyl transferase dUTP nick-end labeling and cleaved caspase-3 showed most cells in the skin inactivated by pressurization to be apoptotic. Consequently, in vivo grafting of apoptosis-induced inactivated skin resulted in successful engraftment and greater dermal cellular density and macrophage infiltration than our existing method. Our finding supports an alternative approach to hydrostatic pressure application.

Improved viability of murine skin flaps using a gelatin hydrogel sheet impregnated with bFGF.

Basic fibroblast growth factor (bFGF) promotes epithelial cell proliferation and angiogenesis but its clinical applications are limited by its short half-life and low retention. Recently developed gelatin hydrogel sheets able to release physiologically active substances in a controlled manner have the potential to overcome these issues. In this study, the effects of gelatin hydrogel sheets impregnated with bFGF on flap survival and angiogenesis were examined in a murine skin flap model. A flap of 1 × 3 cm was generated on the backs of 60 C57BL/6 mice. The mice were divided into five groups (n = 12/group): Group I, untreated; Group II, treated with a gelatin hydrogel sheet impregnated with saline; Group III, treated with bFGF (50 µg) without sheets; Groups IV and V, treated with gelatin hydrogel sheets impregnated with 50 and 100 µg of bFGF, respectively. On the seventh day after surgery, the flap survival area and vascular network were examined and hematoxylin and eosin and von Willebrand factor staining were used for histological examinations. The flap survival areas were significantly larger in Groups IV and V than in other groups. The area of new vessels was significantly larger in Group IV than in the other groups. In the murine skin flap model, gelatin hydrogel sheets impregnated with bFGF promoted angiogenesis and improved flap survival. These findings support the use of bFGF-impregnated gelatin hydrogel sheets for improving ischemic flap survival in clinical settings.

A comparison of the wound healing process after the application of three dermal substitutes with or without basic fibroblast growth factor impregnation in diabetic mice.

Pelnac GplusⓇ, IntegraⓇ, and TerudermisⓇ are approved artificial dermis products in Japan. Previously, we proved that Pelnac GplusⓇ was able to sustain basic fibroblast growth factor (bFGF) and accelerated wound healing by releasing impregnated bFGF. In this study, we impregnated Pelnac GplusⓇ, IntegraⓇ, and TerudermisⓇ with bFGF and compared the binding activity and wound-healing process. We applied bFGF to each material and compared the bFGF concentrations in the surrounding area after 24-h incubation. For the in vivo study, dermal substitutes were impregnated with bFGF and implanted into full-thickness wounds of BKS.Cg-+Leprdb/+Leprdb/Jcl mice. Wounds were evaluated at days 7, 14, and 21 after implantation. The in vitro study showed that bFGF is strongly bound to IntegraⓇ, followed by Pelnac GplusⓇ and TerudermisⓇ. The in vivo study showed that fibroblasts and capillaries had infiltrated into the Pelnac GplusⓇ but not the IntegraⓇ or TerudermisⓇ. Furthermore, long epithelium and wide granulation tissue were formed in the Pelnac GplusⓇ with bFGF group. The TerudermisⓇ with bFGF group had more capillaries than other groups, but only at the base of the wound. The combination of Pelnac GplusⓇ with bFGF may be a novel approach for treating full-thickness skin defects or chronic skin ulcers.

Preparing Activated Platelet-Rich Plasma for Culturing Human Adipose-Derived Stem Cells.

Activated platelet-rich plasma (PRP) prepared from whole blood via centrifugation demonstrated a proliferation-stimulating effect in several kinds of cultured cells, implying a possible use in regenerative medicine. Here, a double-spin method was used to prepare PRP from whole blood. PRP was further activated by autologous thrombin. The platelet count was measured in the activated PRP and the proliferation-stimulating effect in human adipose-derived stem cells (hASCs) was examined. The resulting platelet count was 11.5-times higher in PRP than in whole blood plasma. The proliferation of hASCs was markedly enhanced by incubation with 1% PRP. The described method can be used to reproducibly prepare PRP with a high concentration of platelets. PRP prepared by this method markedly promotes proliferation of hASCs.

High Hydrostatic Pressure Therapy Annihilates Squamous Cell Carcinoma in a Murine Model.

Cutaneous squamous cell carcinoma (cSCC) is one of the most common skin cancers. In the treatment of cSCC, it is necessary to remove it completely, and reconstructive surgery, such as a skin graft or a local or free flap, will be required, depending on the size, with donor-site morbidity posing a burden to the patient. The high hydrostatic pressure (HHP) technique has been developed as a physical method of decellularizing various tissues. We previously reported that HHP at 200 MPa for 10 min could inactivate all cells in the giant congenital melanocytic nevus, and we have already started a clinical trial using this technique. In the present study, we explored the critical pressurization condition for annihilating cSCC cells in vitro and confirmed that this condition could also annihilate cSCC in vivo. We prepared 5 pressurization conditions in this study (150, 160, 170, 180, and 190 MPa for 10 min) and confirmed that cSCC cells were killed by pressurization at ≥160 MPa for 10 min. In the in vivo study, the cSCC cells inactivated by HHP at 200 MPa for 10 min were unable to proliferate after injection into the intradermal space of mice, and transplanted cSCC tissues that had been inactivated by HHP showed a decreased weight at 5 weeks after implantation. These results suggested that HHP at 200 MPa for 10 min was able to annihilate SCC, so HHP technology may be a novel treatment of skin cancer.

Exploration of the Pressurization Condition for Killing Human Skin Cells and Skin Tumor Cells by High Hydrostatic Pressure.

High hydrostatic pressure (HHP) is a physical method for inactivating cells or tissues without using chemicals such as detergents. We previously reported that HHP at 200 MPa for 10 min was able to inactivate all cells in skin and giant congenital melanocytic nevus (GCMN) without damaging the extracellular matrix. We also reported that HHP at 150 MPa for 10 min was not sufficient to inactivate them completely, while HHP at 200 MPa for 10 min was able to inactivate them completely. We intend to apply HHP to treat malignant skin tumor as the next step; however, the conditions necessary to kill each kind of cell have not been explored. In this work, we have performed a detailed experimental study on the critical pressure and pressurization time using five kinds of human skin cells and skin tumor cells, including keratinocytes (HEKas), dermal fibroblasts (HDFas), adipose tissue-derived stem cells (ASCs), epidermal melanocytes (HEMa-LPs), and malignant melanoma cells (MMs), using pressures between 150 and 200 MPa. We pressurized cells at 150, 160, 170, 180, or 190 MPa for 1 s, 2 min, and 10 min and evaluated the cellular activity using live/dead staining and proliferation assays. The proliferation assay revealed that HEKas were inactivated at a pressure higher than 150 MPa and a time period longer than 2 min, HDFas and MMs were inactivated at a pressure higher than 160 MPa and for 10 min, and ASCs and HEMa-LPs were inactivated at a pressure higher than 150 MPa and for 10 min. However, some HEMa-LPs were observed alive after HHP at 170 MPa for 10 min, so we concluded that HHP at a pressure higher than 180 MPa for 10 min was able to inactivate five kinds of cells completely.

Effects of Fibroblast Growth Factor-2 Combined With a Collagen/Gelatin Sponge for Adipogenesis in the Mouse Subcutis.

INTRODUCTION: A collagen/gelatin sponge (CGS) is a new scaffold that promotes wound healing by slowly releasing fibroblast growth factor (FGF)-2. FGF-2 induces mitogenesis, angiogenesis, and adipogenesis. In this study, the adipogenesis-inducing effects of CGS combined with FGF-2 in the subcutis of mice were evaluated. METHODS: Collagens/gelatin sponges (10 × 5 mm) were impregnated with 50 µL of FGF-2 solution (10 or 100 µg/mL). A CGS (Gunze Corp, Osaka, Japan) combined with FGF-2 was implanted subcutaneously into the thoracic region of mice. At 1, 2, 3, and 4 weeks, samples were collected for hematoxylin and eosin staining, von Willebrand factor immunostaining, and perilipin immunostaining to examine adipose tissue localization and angiogenesis. A CGS with only saline solution was prepared as a control. RESULTS: Adipocytes in the collagen fibers appeared at 3 weeks, and a zonal fat layer was noted under the panniculus carnosus at 4 weeks in the FGF-2-combined CGS groups. The fat layer was significantly thicker in the FGF-2 (100 µg/mL) group than in the FGF-2 (10 µg/mL) group. In the control group, no fat pad was newly formed. The number of newly formed vessels in the FGF (10 µg/mL) and (100 µg/mL) groups was significantly greater in the FGF-2 group than in the control group. CONCLUSION: This study presents a promising method to enhance adipogenic effects in the murine subcutis using CGS combined with FGF-2, representing a potential technique for soft tissue reconstruction.