Blood vessels sense dermal stiffness via a novel mechanotransducer, APJ.

Microvascular dysfunction accompanied by a dramatic alteration of stable capillary structure is a major hallmark of numerous age-related diseases. In skin, although the role of angiogenesis during dermal reconstitution is well documented, the functional relevance of the extracellular matrix (ECM) stiffness to vascular remodeling and its molecular mechanisms was poorly understood. Here, we developed an ex vivo 3-dimensional angiogenic model using human fat, revealing that "appropriate" stiffness induces vascular maturation associated with upregulated APJ expression, whereas the overexpression of APJ promotes the formation of large vessels even in the absence of the "appropriate" stiffness. Taken together, APJ could be a novel mechanotransducer that accelerates the maturation of cutaneous blood vessels, leading to the prevention of human skin aging.

Cell-Assisted Lipotransfer for Cosmetic Breast Augmentation: Supportive Use of Adipose-Derived Stem/Stromal Cells.

BACKGROUND: Lipoinjection is a promising treatment but has some problems, such as unpredictability and a low rate of graft survival due to partial necrosis. METHODS: To overcome the problems with lipoinjection, the authors developed a novel strategy known as cellassisted lipotransfer (CAL). In CAL, autologous adiposederived stem (stromal) cells (ASCs) are used in combination with lipoinjection. A stromal vascular fraction (SVF) containing ASCs is freshly isolated from half of the aspirated fat and recombined with the other half. This process converts relatively ASC-poor aspirated fat to ASC-rich fat. This report presents the findings for 40 patients who underwent CAL for cosmetic breast augmentation. RESULTS: Final breast volume showed augmentation by 100 to 200 ml after a mean fat amount of 270 ml was injected. Postoperative atrophy of injected fat was minimal and did not change substantially after 2 months. Cyst formation or microcalcification was detected in four patients. Almost all the patients were satisfied with the soft and natural-appearing augmentation. CONCLUSIONS: The preliminary results suggest that CAL is effective and safe for soft tissue augmentation and superior to conventional lipoinjection. Additional study is necessary to evaluate the efficacy of this technique further.

Progenitor-enriched adipose tissue transplantation as rescue for breast implant complications.

Breast enhancement with artificial implants is one of the most frequently performed cosmetic surgeries but is associated with various complications, such as capsular contracture, that lead to implant removal or replacement at a relatively high rate. For replacement, we used transplantation of progenitor-supplemented adipose tissue (cell-assisted lipotransfer; CAL) in 15 patients. The stromal vascular fraction containing adipose tissue progenitor cells obtained from liposuction aspirates was used to enrich for progenitor cells in the graft. Overall, clinical results were very satisfactory, and no major abnormalities were seen on magnetic resonance imaging or mammogram after 12 months. Postoperative atrophy of injected fat was minimal and did not change substantially after 2 months. Surviving fat volume at 12 months was 155 +/- 50 mL (Right; mean +/- SD) and 143 +/- 80 mL (Left) following lipoinjection from an initial mean of 264 mL. These preliminary results suggest that CAL is a suitable methodology for the replacement of breast implants.

Cell-assisted lipotransfer for facial lipoatrophy: efficacy of clinical use of adipose-derived stem cells.

BACKGROUND: Lipoinjection is a promising treatment, but its efficacy in recontouring facial lipoatrophy remains to be established. OBJECTIVE: The objective was to evaluate the efficacy and adverse effects of lipoinjection and supplementation of adipose-derived stem/stromal cells (ASCs) to adipose grafts. METHODS: To overcome drawbacks of autologous lipoinjection, we have developed a novel strategy called cell-assisted lipotransfer (CAL). In CAL, stromal vascular fraction containing ASCs was freshly isolated from half of an aspirated fat sample and attached to the other half of aspirated fat sample with the fat acting as a scaffold. This process converts relatively ASC-poor aspirated fat into ASC-rich fat. We performed conventional lipoinjection (non-CAL; n=3) or CAL (n=3) on six patients with facial lipoatrophy due to lupus profundus or Parry-Romberg syndrome. RESULTS: All patients obtained improvement in facial contour, but the CAL group had a better clinical improvement score than did the non-CAL patients, although the difference did not reach statistical significance (p=.11). Adipose necrosis was found in one non-CAL case who took perioperative oral corticosteroids. CONCLUSION: Our results suggest that CAL is both effective and safe and potentially superior to conventional lipoinjection for facial recontouring. The authors have indicated no significant interest with commercial supporters.

Influences of centrifugation on cells and tissues in liposuction aspirates: optimized centrifugation for lipotransfer and cell isolation.

BACKGROUND: Although injective autologous fat transplantation is one of the most attractive options for soft-tissue augmentation, problems such as unpredictability and fibrosis resulting from fat necrosis limit its universal acceptance. Centrifugation is one of most common methods for overcoming these difficulties. This study was performed to investigate quantitatively the effects of centrifugation on liposuction aspirates to optimize centrifugal conditions for fat transplantation and isolation of adipose-derived stem cells. METHODS: Liposuction aspirates, obtained from eight healthy female donors, were either not centrifuged or centrifuged at 400, 700, 1200, 3000, or 4200 g for 3 minutes. The volumes of the oil, adipose, and fluid portions and numbers of blood cells and adipose-derived cells in each portion were examined. The processed adipose tissues (1 ml) were injected into athymic mice, and grafts were harvested and weighed at 4 weeks. Morphologic alterations were observed using light and scanning electron microscopy. RESULTS: Centrifugation concentrated adipose tissues and adipose-derived stem cells in the adipose portion and partly removed red blood cells from the adipose portion. Centrifugation at more than 3000 g significantly damaged adipose-derived stem cells. Centrifugation enhanced graft take per 1 ml centrifuged adipose but reduced calculated graft take per 1 ml adipose before centrifugation. CONCLUSIONS: Excessive centrifugation can destroy adipocytes and adipose-derived stem cells, but appropriate centrifugation concentrates them, resulting in enhanced graft take. The authors tentatively recommend 1200 g as an optimized centrifugal force for obtaining good short- and long-term results in adipose transplantation.

Preserved proliferative capacity and multipotency of human adipose-derived stem cells after long-term cryopreservation.

BACKGROUND: Human adipose-derived stem (stromal) cells are promising as a regenerative therapy tool for defective tissues of mesenchymal lineage, including fat, bone, and cartilage, and blood vessels. In potential future clinical applications, adipose-derived stem cell cryopreservation could be an indispensable fundamental technology, as has occurred in other fields involving cell-based therapies using hematopoietic stem cells and umbilical cord blood cells. METHODS: The authors examined the proliferative capacity and multipotency of human adipose-derived stem cells isolated from lipoaspirates of 18 patients in total before and after a 6-month cryopreservation following their defined protocol. Proliferative capacity was quantified by measuring doubling time in cell culture, and multipotency was examined with differentiation assays for chondrogenic, osteogenic, and adipogenic lineages. In addition, expression profiles of cell surface markers were determined by flow cytometry and compared between fresh and cryopreserved adipose-derived stem cells. RESULTS: Cryopreserved adipose-derived stem cells fully retained the potential for differentiation into adipocytes, osteoblasts, and chondrocytes and for proliferative capacity. Flow cytometric analyses revealed that surface marker expression profiles remained constant before and after storage. CONCLUSIONS: Adipose-derived stem cells can be cryopreserved at least for up to 6 months under the present protocol without any loss of proliferative or differentiation potential. These results ensure the availability of autologous banked adipose-derived stem cells for clinical applications in the future.

Cell-assisted lipotransfer for cosmetic breast augmentation: supportive use of adipose-derived stem/stromal cells.

BACKGROUND: Lipoinjection is a promising treatment but has some problems, such as unpredictability and a low rate of graft survival due to partial necrosis. METHODS: To overcome the problems with lipoinjection, the authors developed a novel strategy known as cell-assisted lipotransfer (CAL). In CAL, autologous adipose-derived stem (stromal) cells (ASCs) are used in combination with lipoinjection. A stromal vascular fraction (SVF) containing ASCs is freshly isolated from half of the aspirated fat and recombined with the other half. This process converts relatively ASC-poor aspirated fat to ASC-rich fat. This report presents the findings for 40 patients who underwent CAL for cosmetic breast augmentation. RESULTS: Final breast volume showed augmentation by 100 to 200 ml after a mean fat amount of 270 ml was injected. Postoperative atrophy of injected fat was minimal and did not change substantially after 2 months. Cyst formation or microcalcification was detected in four patients. Almost all the patients were satisfied with the soft and natural-appearing augmentation. CONCLUSIONS: The preliminary results suggest that CAL is effective and safe for soft tissue augmentation and superior to conventional lipoinjection. Additional study is necessary to evaluate the efficacy of this technique further.

Influences of preservation at various temperatures on liposuction aspirates.

BACKGROUND: Aspirated fat is not only a filler material but also an abundant source of adipose-derived stem cells. The aim of this study was to assess degeneration of aspirated fat during preservation and optimize the preservation method for lipoaspirates. METHODS: Aspirated fat was preserved at room temperature for 1, 2, 4, and 24 hours (n = 10 each); at 4 degrees C for 1, 2, and 3 days (n = 14 each); or at -80 degrees C for 1 month (n = 3). Morphologic changes were assessed with scanning electron microscopy. Adipose-derived stem cell yield was measured after 1 week of culture. For aspirated fat preserved at room temperature, damaged adipocytes were assessed by measuring the oil volume ratio after centrifugation (n = 6) and glycerol-3-phosphate-dehydrogenase activity in washing solution (n = 4). Cell surface marker expression was examined by flow cytometry (n = 3). RESULTS: Although the scanning electron microscopic assay indicated no remarkable anatomical changes based on preservation methods, oil volume significantly increased in fat preserved at room temperature for 4 hours. Adipose-derived stem cell yield was significantly reduced by preservation at room temperature for 24 hours and by preservation at 4 degrees C for 2 or 3 days. Flow cytometric analysis suggested that the biological properties of adipose-derived stem cells did not significantly change at 4 degrees C up to 3 days. The cells were isolated from cryopreserved fat, but the yield was much less than that from fresh aspirated fat. CONCLUSIONS: Aspirated fat should be transplanted as quickly as possible if it is preserved at room temperature. For adipose-derived stem cell isolation, aspirated fat can be stored or transported overnight if it is preserved at 4 degrees C without adipose-derived stem cell yield loss or changes in biological properties.

A clinical trial of topical bleaching treatment with nanoscale tretinoin particles and hydroquinone for hyperpigmented skin lesions.

BACKGROUND: Although combined use of tretinoin (all-trans-retinoic acid; atRA) and hydroquinone improves various hyperpigmented lesions, the pharmacologic instability of atRA and atRA-induced irritant dermatitis are difficult unsolved problems. OBJECTIVE: The objective was to evaluate the efficacy and adverse effects of a newly formulated gel containing inorganic-coated atRA nanoscale particles (nano-atRA gel). METHODS: Nano-atRA gel was used in our two-phased bleaching protocol: 5% hydroquinone and 7% lactic acid ointment were used along with nano-atRA gel in the bleaching phase (2-8 weeks), and 5% hydroquinone and 7% ascorbic acid ointment were used alone during the healing phase (4-8 weeks). Eighty-four patients with facial hyperpigmented lesions were enrolled in this study, and 77 of them (88 lesions) followed up for more than 10 weeks were analyzed. RESULTS: Hyperpigmentation was improved in 84 of 88 lesions (95.5%) after a mean treatment period of 14.3 weeks and was almost eliminated in 52 lesions (59.1%). Nano-atRA gel caused exfoliation and scaling similar to that seen with conventional atRA gel, whereas the erythema seen in the bleaching phase appeared to be weaker. CONCLUSION: Nano-atRA gel can improve hyperpigmentation to a similar extent as conventional atRA gel. It also induces irritant dermatitis, but with less erythema.

Anti-androgenic therapy using oral spironolactone for acne vulgaris in Asians.

BACKGROUND: Few studies have addressed anti-androgenic therapy using oral spironolactone for acne in Asians. Obtaining this race-specific information is important because Westerners and Asians respond differently to hormone therapy. This study aimed to examine the efficacy and safety of oral spironolactone used to treat acne in Asians. METHODS: Spironolactone (initial dose, 200 mg/day) was administered orally to 139 Japanese patients (116 females and 23 males) with acne. Serum laboratory data, including various hormones and electrolytes, were examined for 25 of the subjects. RESULTS: Most of the female patients who completed the 20-week regimen exhibited excellent improvement (evaluated by a photographic grading scale), although some discontinued treatment because of menstrual disturbances or other reasons. The treatment was less efficacious for the males than for the females, and because gynecomastia developed in three male patients, spironolactone treatment for males was stopped. Examination of the serum of 25 patients did not identify any toxicity associated with the treatment. Drug eruptions and edema in the lower extremities were each seen in three patients. CONCLUSION: Oral spironolactone is effective and safe for the treatment of acne in Asian females, and can be a good option for severe, recurring, and widespread types of the condition.

Repeated treatment protocols for melasma and acquired dermal melanocytosis.

BACKGROUND AND OBJECTIVE: Melasma and acquired dermal melanocytosis (ADM; acquired bilateral nevus of Ota-like macules) are both seen most commonly symmetrically on the face of women with darker skin and are also known as difficult conditions to treat. METHODS: Our topical bleaching protocol with 0.1 to 0.4% tretinoin gel and 5% hydroquinone was performed repeatedly (1-3 times) for melasma (n=163), and a combination treatment with topical bleaching and Q-switched ruby (QSR) laser was performed repeatedly (1-3 times) for ADM (n=62). RESULTS: There is a significant correlation between clinical results (clearance of pigmentation) and the number of sessions in both melasma (p=.019) and ADM (p<.0001). CONCLUSION: The repeated treatment protocol for melasma and ADM showed successful clinical results compared with conventional ones, and they may be applied to other pigment conditions. It may be better that epidermal and dermal pigmentations are treated separately, especially in dark-skinned people who are more likely to suffer postinflammatory hyperpigmentation after inflammation-inducing therapies.

Characterization of freshly isolated and cultured cells derived from the fatty and fluid portions of liposuction aspirates.

Liposuction aspirates (primarily saline solution, blood, and adipose tissue fragments) separate into fatty and fluid portions. Cells isolated from the fatty portion are termed processed lipoaspirate (PLA) cells and contain adipose-derived adherent stromal cells (ASCs). Here we define cells isolated from the fluid portion of liposuction aspirates as liposuction aspirate fluid (LAF) cells. Stromal vascular fractions (SVF) were isolated separately from both portions and characterized under cultured and non-cultured conditions. A comparable number of LAF and PLA cells were freshly isolated, but fewer LAF cells were adherent. CD34+ CD45- cells from fresh LAF isolates were expanded by adherent culture, suggesting that LAF cells contain ASCs. Although freshly isolated PLA and LAF cells have distinct cell surface marker profiles, adherent PLA and LAF cells have quite similar characteristics with regard to growth kinetics, morphology, capacity for differentiation, and surface marker profiles. After plating, both PLA and LAF cells showed significant increased expression of CD29, CD44, CD49d, CD73, CD90, CD105, and CD151 and decreased expression of CD31 and CD45. Multicolor FACS analysis revealed that SVF are composed of heterogeneous cell populations including blood-derived cells (CD45+), ASCs (CD31- CD34+ CD45- CD90+ CD105- CD146-), endothelial (progenitor) cells (CD31+ CD34+ CD45- CD90+ CD105low CD146+), pericytes (CD31- CD34- CD45- CD90+ CD105- CD146+), and other cells. After plating, ASCs showed a dramatic increase in CD105 expression. Although some adherent ASCs lost CD34 expression with increasing culture time, our culture method maintained CD34 expression in ASCs for at least 10-20 weeks. These results suggest that liposuction-derived cells may be useful and valuable for cell-based therapies.

Cell-assisted lipotransfer: supportive use of human adipose-derived cells for soft tissue augmentation with lipoinjection.

Injective transfer of autologous aspirated fat is a popular option for soft tissue augmentation, but several issues require attention, including unpredictability and a low survival rate due to partial necrosis. In this study, histologic features and yield of adipose-derived stromal (stem) cells (ASCs) were compared between human aspirated fat and excised whole fat. Aspirated fat contained fewer large vascular structures, and ASC yield was lower in aspirated fat. Aspirated fat was transplanted subcutaneously into severe combined immunodeficiency mice with (cell-assisted lipotransfer; CAL) or without (non-CAL) vascular stromal fractions containing ASCs isolated from adipose tissue. The CAL fat survived better (35% larger on average) than non-CAL fat, and microvasculature was detected more prominently in CAL fat, especially in the outer layers. DiI-labeled vascular stromal fraction cells were found between adipocytes and in the connective tissue in CAL fat, and some of these cells were immunopositive for von Willebrand factor, suggesting differentiation into vascular endothelial cells. Another experiment that used vascular stromal fractions taken from green fluorescent protein rats also suggested that ASCs differentiated into vascular endothelial cells and contributed to neoangiogenesis in the acute phase of transplantation. These findings may partly explain why transplanted aspirated fat does not survive well and suggest clinical potential of the CAL method for soft tissue augmentation.

Simultaneous reconstruction of breast and well-projected nipple after expansion of mammary skin.

The problem of postoperative reduction of projecting reconstructed nipples remains to be resolved. To this end we did a clinical study of reconstructing the nipple at the same time as the breast. A tissue-expander was placed under the skin of the breast at the first operation, and then the breast and nipple were reconstructed at the second. A nipple was reconstructed using a dermal-fat flap harvested from the myocutaneous flaps used for reconstruction of the breast. A small hole was made in the corresponding site of the skin of the breast, and the reconstructed nipple was projected through the hole. This method was used in 8 cases. This method is useful in reconstructing a breast without a pad of skin and a projected nipple simultaneously. Its disadvantages are the relatively weak blood supply of the flaps, and difficulty in calculating the position of the nipple. The procedure may be beneficial for selected cases.

Angle-splitting ostectomy followed by face lift for elderly patients with prominent mandibular angles.

A prominent mandibular angle is a relatively common aesthetic problem among Asians, and the reduction angle-splitting ostectomy is now becoming a very popular procedure in Asian countries. Although this operation is usually performed on young patients, the same aesthetic demands are also seen in the elderly. In this report, the authors describe their experience with angle-splitting ostectomies followed by face lifts in three patients older than 50 years. The angle-splitting procedure was the same as that performed in young patients, and clinical results were assessed with photographs and three-dimensional computed tomographic scans. The facial contours after angle-splitting ostectomy were satisfactory, but the patients showed postoperative redundancy of the skin, especially along the jaw line, because of the loss of bony protrusion laterally. Therefore, the patients underwent subsequent superficial musculoaponeurotic system cheek lifts. The final aesthetic results were satisfactory in all cases. When surgeons want to perform the angle-splitting ostectomy safely and effectively on the elderly, they should be aware of the risks and indications specific for elderly patients, and a multidisciplinary support system should be available. Subsequent face lifts can improve skin redundancy and lead to better cosmetic results.

Combined therapy using Q-switched ruby laser and bleaching treatment with tretinoin and hydroquinone for acquired dermal melanocytosis.

BACKGROUND AND OBJECTIVE: Acquired dermal melanocytosis (ADM; acquired bilateral nevus of Ota-like macules) is known for its recalcitrance compared with Nevus of Ota, and we assume that one of the reasons is a higher rate and degree of postinflammatory hyperpigmentation (PIH) seen after laser treatments. METHODS: Topical bleaching treatment with 0.1% tretinoin aqueous gel and 5% hydroquinone ointment containing 7% lactic acid was initially performed (4 to 6 weeks) to discharge epidermal melanin. Subsequently, Q-switched ruby (QSR) laser was irradiated to eliminate dermal pigmentation. Both steps were repeated two to three times until patient satisfaction was obtained (usually at a 2-month interval for laser sessions). This treatment was performed in 19 patients with ADM. Skin biopsy was performed in six cases at baseline, after the bleaching pretreatment, and at the end of treatment. RESULTS: All patients showed good to excellent clearing after two to three sessions of QSR laser treatments. The total treatment period ranged from 3 to 13 (mean of 8.3) months. PIH was observed in 10.5% of the cases. Histologically, epidermal hyperpigmentation was observed in all specimens and was dramatically improved by the topical bleaching pretreatment. CONCLUSION: QSR laser combined with the topical bleaching pretreatment appeared to treat ADM consistently with a low occurrence rate of PIH and lessen the number of laser sessions and total treatment period and may also be applied to any other lesions with both epidermal and dermal pigmentation.