Giulia Lo Russo

Autologous lipofilling: coenzyme Q10 can rescue adipocytes from stress-induced apoptotic death.

Background: Autologous fat transplantation (or lipofilling) is an excellent technique for correction of cosmetic defects. The success of the procedure relies strongly on the techniques of harvesting and transferring viable adipocytes. The purpose of this study was to evaluate effects of two harvesting methods and coenzyme Q10 on the viability and apoptotic death of adipocytes collected for autologous lipofilling. Methods: Human adipose tissue from six patients was collected by Luer-Lok syringe according to Coleman’s technique or by means of an aspirator with a 680-mmHg vacuum. Half of each sample collected using Coleman’s technique was treated with 10 &mgr;M Coenzyme Q10, and the other half served as untreated control. Viability and apoptosis were assessed by immunoenzymatic, biochemical, and morphological methods. Results: The harvesting of adipose tissue by aspirator reduced the viability and increased apoptotic death significantly more than harvesting tissue using Coleman’s technique. Biochemical and morphological analyses confirmed that treatment of adipose tissue with coenzyme Q10 reduced and even inhibited apoptotic death of harvested adipocytes. Conclusion: Coenzyme Q10 can rescue adipocytes from stress-induced apoptotic death.

Cutaneous ciliated cyst: A case report with immunohistochemical evidence for dynein in ciliated cells

Cutaneous ciliated cysts (CCCs) are rare benign lesions predominantly occurring in the lower limbs of young women. We observed such a lesion in the perineal region of a 12-year-old girl. The histogenetic interpretation of CCCs is controversial. The similarity of the epithelial lining of the cyst to the salpingeal epithelium supports the hypothesis of a Müllerian heterotopia. Strong dynein positivity observed immunohistochemically in the apical portion of CCC lining cells suggests the integrity of the ciliary apparatus. There was a marked similarity between a CCC and normal salpingeal epithelium in the mode of staining for dynein, whereas the dynein reactivity of bronchial epithelial cells showed larger cytoplasmic aggregates of positive material in proximity to the nucleus. No immunohistochemical staining for estrogen or progesterone receptors was identified in our observation, although this finding could have supported the Müllerian histogenetic hypothesis. To the best of our knowledge, this is the first time that antidynein immunohistochemistry has been applied to paraffin-embedded samples from human surgical pathology.

Oncocytoma of the nasal cavity: report of a case and review of the literature

The authors present a case of oncocytoma of the nasal cavity in a 60-year-old woman. The tumour shows the same histological and ultrastructural characteristics of oncocytomas arising in minor salivary glands of other sites and major salivary glands. The authors stress that the biological behaviour of oncocytomas of the nasal cavity seems related to their local growth rather than to the cytological characteristics of the tumour. The extreme rarity of this tumour in the nasal cavity is particularly emphasized. The few cases reported previously in literature are briefly reviewed.

MSCs seeded on bioengineered scaffolds improve skin wound healing in rats.

Growing evidence has shown the promise of mesenchymal stromal cells (MSCs) for the treatment of cutaneous wound healing. We have previously demonstrated that MSCs seeded on an artificial dermal matrix, Integra (Integra Lifesciences Corp., Plainsboro, NJ) enriched with platelet‐rich plasma (Ematrix) have enhanced proliferative potential in vitro as compared with those cultured on the scaffold alone. In this study, we extended the experimentation by evaluating the efficacy of the MSCs seeded scaffolds in the healing of skin wounds in an animal model in vivo. It was found that the presence of MSCs within the scaffolds greatly ameliorated the quality of regenerated skin, reduced collagen deposition, enhanced reepithelization, increased neo‐angiogenesis, and promoted a greater return of hair follicles and sebaceous glands. The mechanisms involved in these beneficial effects were likely related to the ability of MSCs to release paracrine factors modulating the wound healing response. MSC‐seeded scaffolds, in fact, up‐regulated matrix metalloproteinase 9 expression in the extracellular matrix and enhanced the recruitment of endogenous progenitors during tissue repair. In conclusion, the results of this study provide evidence that the treatment with MSC‐seeded scaffolds of cutaneous wounds contributes to the recreation of a suitable microenvironment for promoting tissue repair/regeneration at the implantation sites.

Adipofascial Anterolateral Thigh Flap Safety: Applications and Complications

Background A thinned anterolateral thigh (ALT) flap is often harvested to achieve optimal skin resurfacing. Several techniques have been described to thin an ALT flap including an adipocutaneous flap, an adipofascial flap and delayed debulking. Methods By systematically reviewing all of the available literature in English and French, the present manuscript attempts to identify the common surgical indications, complications and donor site morbidity of the adipofascial variant of the ALT flap. The studies were identified by performing a systematic search on Medline, Ovid, EMBASE, the Cochrane Database of Systematic Reviews, Current Contents, PubMed, Google, and Google Scholar. Results The study selection process was adapted from the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement, and 15 articles were identified using the study inclusion criteria. These articles were then reviewed for author name(s), year of publication, flap dimensions and thickness following defatting, perforator type, type of transfer, complications, thinning technique, number of cases with a particular area of application and donor site morbidity. Conclusions The adipofascial variant of the ALT flap provides tissue to fill large defects and improve pliability. Its strong and safe blood supply permits adequate immediate or delayed debulking without vascular complications. The presence of the deep fascia makes it possible to prevent sagging by suspending and fixing the flap for functional reconstructive purposes (e.g., the intraoral cavity). Donor site morbidity is minimal, and thigh deformities can be reduced through immediate direct closure or liposuction and direct closure. A safe blood supply was confirmed by the rate of secondary flap debulking.

Outcome of Patients with Melanoma and Histologically Negative Sentinel Lymph Nodes: One Institution’s Experience

Background: Since its introduction by Morton in 1992, sentinel lymph node (SLN) biopsy has become a standard procedure in the staging and treatment of primary melanoma and clinically negative regional lymph nodes. The primary aims of this procedure are to ascertain the individual lymphatic drainage patterns of primary tumors towards 1 or more different lymph node basins and to identify patients with micrometastatic lymphatic disease for selective lymphadenectomy. The aim of our study was to evaluate over time a cohort of patients who, having undergone SLN treatment, were found negative for metastases using routine histopathological and immunohistochemical analyses. Methods: We studied 102 consecutive patients who underwent intraoperative lymphatic mapping at the Department of Plastic Surgery, University of Florence, Italy, for cutaneous melanoma and were found negative for metastatic melanoma in their SLNs using routine histopathological and immunohistochemical techniques. Results: Of 102 patients with 103 cutaneous melanomas that underwent SLN resection and proved histologically negative to metastasis in that site, 15 patients (14.7%) developed melanoma recurrence during follow-up. Conclusions: The diagnostic and prognostic value of the absence of melanoma metastases in SLNs may be limited and not particularly significant, since satellite and in-transit metastases or direct distant metastases will not be detected and hematogenous spread may already have begun at the time of intervention.

Histological and Ultrastructural Effects of Ultrasound-induced Cavitation on Human Skin Adipose Tissue

Background: In aesthetic medicine, the most promising techniques for noninvasive body sculpturing purposes are based on ultrasound-induced fat cavitation. Liporeductive ultrasound devices afford clinically relevant subcutaneous fat pad reduction without significant adverse reactions. This study aims at evaluating the histological and ultrastructural changes induced by ultrasound cavitation on the different cell components of human skin. Methods: Control and ultrasound-treated ex vivo abdominal full-thickness skin samples and skin biopsies from patients pretreated with or without ultrasound cavitation were studied histologically, morphometrically, and ultrastructurally to evaluate possible changes in adipocyte size and morphology. Adipocyte apoptosis and triglyceride release were also assayed. Clinical evaluation of the effects of 4 weekly ultrasound vs sham treatments was performed by plicometry. Results: Compared with the sham-treated control samples, ultrasound cavitation induced a statistically significant reduction in the size of the adipocytes (P < 0.001), the appearance of micropores and triglyceride leakage and release in the conditioned medium (P < 0.05 at 15 min), or adipose tissue interstitium, without appreciable changes in microvascular, stromal, and epidermal components and in the number of apoptotic adipocytes. Clinically, the ultrasound treatment caused a significant reduction of abdominal fat. Conclusions: This study further strengthens the current notion that noninvasive transcutaneous ultrasound cavitation is a promising and safe technology for localized reduction of fat and provides experimental evidence for its specific mechanism of action on the adipocytes.

Sentinel Lymph Nodes in Melanoma Patients: Evaluating the Evidence

Identification of patients with occult melanoma metastases is considered important for accurate staging, treatment planning, and prognosis. One potential bias in the SLNB theory is that all positive SLNs inevitably progress to overt nodal disease if lymphadenectomy is not performed. There is now strong evidence that this is not the case and that some micrometastases in the SLN (often small aggregates of neoplastic cells in the peripheral sinuses) are destined either for destruction by host-immune processes or for dormancy [5–12] . Moreover, there is no final proof that SLNB, with selective complete lymph node dissection for a tumor-involved SLN, improves survival of melanoma patients [3, 4] . Moreover, it has been reported that approximately 13% of SLN-negative patients will experience a disease progression within 3 years from surgery [6, 7] . Of greater concern is the risk of false positivity leading to unnecessary lymphadenectomy, an issue which is rarely discussed [6] . So far, the SLN procedure has been mainly used to stage melanoma patients and to select patients for adjuvant and/or experimental therapies at an early stage. Such data are extremely significant and cannot be dismissed as normal figures of false negatives peculiar to one procedure. Furthermore, false-negative results can Although evaluation of the sentinel lymph node (SLN) status is included in the new American Joint Committee on Cancer staging system [1, 2] , there is no final evidence that the SLN procedure in melanoma patients influences clinical outcome [3, 4] . Since its introduction by Morton [5] in 1992, SLN biopsy (SLNB) has become a standard procedure in the staging and treatment of primary melanomas with a tumor thickness 6 1 mm (or ! 1 mm if ulcerated or Clark’s level 6 IV) and clinically negative regional lymph nodes. Therefore, lymphatic mapping with sentinel lymphadenectomy has been widely adopted by the surgical oncology community as a standard of care in managing patients with primary cutaneous melanoma, without rigorous prospective clinical trials. The SLN concept is based on the theory of an orderly progression of tumor cells in the initial phase of the metastatic process within the lymphatic system. It assumes that early lymphatic metastases, if present, are initially always found within the first tumor-draining lymph node, the SLN. The primary aims of this procedure are to ascertain the individual lymphatic drainage patterns of primary tumors towards one or more different lymph node basins and to identify patients with micrometastatic lymphatic disease for selective lymphadenectomy. Received: March 29, 2007 Accepted: May 26, 2007 Published online: September 14, 2007

A novel combined surgical approach to head and neck dermatofibrosarcoma protuberans

INTRODUCTION The surgical management of dermatofibrosarcoma protuberans has historically been a challenge, particularly in the head and neck and other aesthetic areas. The current priority is to achieve local oncologic control and a good reconstructive outcome. Here, we present our experience using a novel combined approach with a dermal regenerative template, sub-atmospheric pressure and skin graft. MATERIALS AND METHODS Five patients presenting at the Department of Plastic and Reconstructive Surgery of the University of Florence between January 2010 and October 2011 were included in the study following Institutional Review Board approval. All patients underwent a wide local excision of dermatofibrosarcoma protuberans affecting the head and neck. RESULTS The combined approach using the negative-pressure device, a dermal regenerative template and skin grafting proved effective in the management of this type of sarcoma with a good aesthetic and functional outcome, particularly on the neck or the supraclavicular region. CONCLUSION Although the present multi-step technique requires patient compliance, it results in good local oncologic control of the resection margins. It is possible to perform a wider excision in the event of positive margins without interfering with the last reconstructive outcome.

Bio-Engineered Mesenchymal Stromal Cell (MSCs) Grafts for Skin Repair/ Regeneration - Preclinical Aspects

There is growing evidence showing the promise of mesenchymal stromal cells (MSCs) for the treatment of cutaneous wound healing. In a previous study, we have shown that MSCs seeded on an artificial dermal matrix, Integra ® enriched with platelet rich plasma (Ematrix TM ) have enhanced proliferative potential as compared to those cultured on the scaffold alone. In this study, we wanted to extend the experimentation by evaluating the efficacy of the MSCs bioengineered scaffolds in the healing of skin wounds. To this purpose, full-thickness skin defects were created on the dorsum of rats and covered with (a) Integra®, (b) Ematrix TM , (c) Integra® plus MSCs, (d) and Ematrix TM plus MSCs, or (e) left to heal spontaneously (control). It was found that the presence of MSCs within the scaffolds significantly accelerated wound healing and greatly ameliorated the quality of regenerated skin; it reduced collagen deposition, enhanced re-epithelization, increased neo-angiogenesis and promoted a greater return of hair follicles and sebaceous glands. No evidence of MSC transdifferentiation was seen, suggesting that the therapeutic benefits of the seeded cells were primarily due to their ability to release paracrine factors modulating the wound healing response. Of interest, the presence of MSCs within the scaffolds enhanced the recruitment and differentiation of the endogenous tissue progenitors, suggesting the recreation at the implantation sites of a suitable microenvironment for promoting regeneration. In conclusion, the results of this study provide strong evidence that the treatment with MSC-seeded scaffolds represents an attractive approach for augmenting the regenerative potential and enhancing cutaneous wound healing.