Promotion of Survival and Engraftment of Transplanted Adipose Tissue-Derived Stromal and Vascular Cells by Overexpression of Manganese Superoxide Dismutase.

Short-term persistence of transplanted cells during early post-implant period limits clinical efficacy of cell therapy. Poor cell survival is mainly due to the harsh hypoxic microenvironment transplanted cells face at the site of implantation and to anoikis, driven by cell adhesion loss. We evaluated the hypothesis that viral-mediated expression of a gene conferring hypoxia resistance to cells before transplant could enhance survival of grafted cells in early stages after implant. We used adipose tissue as cell source because it consistently provides high yields of adipose-tissue-derived stromal and vascular cells (ASCs), suitable for regenerative purposes. Luciferase positive cells were transduced with lentiviral vectors expressing either green fluorescent protein as control or human manganese superoxide dismutase (SOD2). Cells were then exposed in vitro to hypoxic conditions, mimicking cell transplantation into an ischemic site. Cells overexpressing SOD2 displayed survival rates significantly greater compared to mock transduced cells. Similar results were also obtained in vivo after implantation into syngeneic mice and assessment of cell engraftment by in vivo bioluminescent imaging. Taken together, these findings suggest that ex vivo gene transfer of SOD2 into ASCs before implantation confers a cytoprotective effect leading to improved survival and engraftment rates, therefore enhancing cell therapy regenerative potential.

Interdomal-columellar myolipoma: discovered at rhinoplasty.

UNLABELLED: Tumors of the adipose tissue located in the nose are extremely rare. This report draws attention to the first case of nasal tip-columellar myolipoma described in the literature. During an open rhinoplasty, the authors elevated the columellar flap and approached the tip, where they found a misdiagnosed 2 × 1 cm fatty mass wedged into the interdomal space continuing between the middle and medial cruses of the alar cartilages, which were laterally dislocated. The mass was excised by sharp and blunt dissection, and histopathologic examination resulted in a diagnosis of myolipoma. This report presents a novel location of myolipoma discovered incidentally. Its unexpected presence affected the surgeons in their operational decisions and forced them to make a rapid change in their surgical plan. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Harvest of superficial layers of fat with a microcannula and isolation of adipose tissue-derived stromal and vascular cells.

BACKGROUND: Adipose tissue is a source of stromal and vascular cells suitable for regenerative medical applications. Cell recovery depends on several factors, including the characteristics of the cannula used to harvest tissue. OBJECTIVES: The authors assess whether aspiration of superficial layers of adipose tissue performed with a microcannula, rather than a standard cannula, allows for improved isolation of stromal and vascular cells, and they evaluate the angiogenic potential of the isolated cells in vitro and in vivo. METHODS: Adipose-derived stromal and stem cells (ADSC) were collected from the lipoaspirate of the abdomen and hip regions of 6 healthy female donors. For adipose tissue harvest, several options were compared: (1) a rounded-tip cannula with a length of 170 mm, a diameter of 3 mm, and a single elliptic suction port on the side near its distal end (port diameter: 3 × 9 mm) or (2) a rounded-tip infiltration cannula with a length of 170 mm, a diameter of 2 mm, and 5 round ports placed spirally along the sides of the distal cannula shaft (each port diameter: 1 mm) (Shipper Medical Technologies Corporation, Centennial, Colorado). Isolated cells were characterized for (1) expression of the endothelial specific marker CD31 by immunohistochemical and cytofluorimetric analyses and (2) tubular-like structure formation using a 3-dimensional angiogenesis assay on Matrigel. Human ADSC were transduced to express firefly luciferase as a marker suitable for bioluminescent tracking and transplantation studies into immunosuppressed mice were performed. RESULTS: ADSC yield was determined to be significantly higher in samples collected with the microcannula (P = .04). Moreover, isolated cells acquired typical endothelial-like morphology in vitro, formed capillary-like structures, and expressed the distinctive endothelial cell marker CD31. Cells implanted into immunosuppressed mice persisted for several weeks in areas undergoing neovascularization. CONCLUSIONS: These results suggest that aspiration of adipose tissue with a microcannula can be a minimally invasive method to obtain clinically relevant numbers of stromal and vascular cells useful for autologous transplant procedures and for promoting tissue regeneration and neovascularization.

AIRMESS - Academy of International Regenerative Medicine & Surgery Societies: recommendations in the use of platelet-rich plasma (PRP), autologous stem cell-based therapy (ASC-BT) in androgenetic alopecia and wound healing.

INTRODUCTION: Many investigations showed that platelet-rich plasma (PRP), human follicles stem cells (HFSCs), and adipose-derived stem cells (ASCs), considered autologous stem cell-based therapy (ASC-BT), are effective for hair regrowth (HR) in patients affected by androgenetic alopecia and for wound healing (WH). The aim of this article is to analyze the in vitro and in vivo impact of different PRP, HFSCs, and ASCs preparation methods on HR and in WH. AREAS COVERED: The analyzed data intended to clarify the molecular mechanism in which PRP, HFSCs, and ASCs are involved, the clinical use and related indications, fully respecting the European rules. Comparative studies between different systems of PRP, HFSCs, and ASCs preparation revealed differences in terms of HR and WH. EXPERT OPINION: Despite a lack of standardized protocols, there is convincing evidence with objective measurement modalities that display positive outcomes of ASC-BT in HR and WH.

Regenerative medicine approaches for the management of respiratory tract fistulas.

Respiratory tract fistulas (or fistulae) are abnormal communications between the respiratory system and the digestive tract or the adjacent organs. The origin can be congenital or, more frequently, iatrogenic and the clinical presentation is heterogeneous. Respiratory tract fistulas can lead to severely reduced health-related quality of life and short survival. Therapy mainly relies on endoscopic surgical interventions but patients often require prolonged hospitalization and may develop complications. Therefore, more conservative regenerative medicine approaches, mainly based on lipotransfer, have also been investigated. Adipose tissue can be delivered either as unprocessed tissue, or after enzymatic treatment to derive the cellular stromal vascular fraction. In the current narrative review, we provide an overview of the main tissue/cell-based clinical studies for the management of various types of respiratory tract fistulas or injuries. Clinical experience is limited, as most of the studies were performed on a small number of patients. Albeit a conclusive proof of efficacy cannot be drawn, the reviewed studies suggest that grafting of adipose tissue-derived material may represent a minimally invasive and conservative treatment option, alternative to more aggressive surgical procedures. Knowledge on safety and tolerability acquired in prior studies can lead to the design of future, larger trials that may exploit innovative procedures for tissue processing to further improve the clinical outcome.

Regenerative application of stromal vascular fraction cells enhanced fat graft maintenance: clinical assessment in face rejuvenation.

OBJECTIVES: The aim of this study was to evaluate the safety and efficacy of the use of FG-SVFs in face rejuvenation for esthetic improvement. METHODS: 33 female patients affected by face's soft-tissue defects with loss of volume, study group (SG), were treated with FG-SVFs, comparing results with a control group (CG) (n = 30) treated with fat graft not enhanced (FG). Clinical evaluation, a photographic assessment, magnetic resonance imaging (MRI), and ultrasound (US) were performed. Post-operative follow-up was performed at 1, 3, 7, 12, 24, 48, weeks, and then annually. RESULTS: SG patients showed 61% maintenance of the contour restoring and of volume after 3 years compared with the CG treated with FG, who showed 31% maintenance. 60.7% (n = 20) of SG patients, presented an increase of 6.6 mm in the soft tissue volume after 36 months, which was reported in only 33,3% (n = 10) of the CG. Volumetric persistence in the SG was higher than that in the CG (p <. 0001 vs. CG). MRI and US moreover confirmed the absence of important side effects, as fat necrosis, and cytosteatonecrotic areas. CONCLUSIONS: The use of FG-SVFs was safe and effective in this series of a case treated.

Treatment of Nasal Aging and Possible Long-term Nasal Enhancement with Fat Using Injectable Tissue Replacement.

This article addresses nasal aging using a minimally invasive procedure with an algorithm that includes the diagnosis of a patient's nose within the context of the individual's facial anatomy. Pyriform augmentation for the elevation of tip projection and columellar strutting with injectable fat are, to our knowledge, applied for the first time as important steps in minimally invasive rhinoplasties. This procedure offers patients with more subtle nasal changes with aging or other causes to be treated with potentially long-lasting improvements, which can be easily retreated. New techniques using injectable liquid cartilage grafts are being explored.

Intraoperative Strategies for Minimal Manipulation of Autologous Adipose Tissue for Cell- and Tissue-Based Therapies: Concise Review.

The stromal vascular fraction (SVF) is a heterogeneous population of stem/stromal cells isolated from perivascular and extracellular matrix (ECM) of adipose tissue complex (ATC). Administration of SVF holds a strong therapeutic potential for regenerative and wound healing medicine applications aimed at functional restoration of tissues damaged by injuries or chronic diseases. SVF is commonly divided into cellular stromal vascular fraction (cSVF) and tissue stromal vascular fraction (tSVF). Cellular SVF is obtained from ATC by collagenase digestion, incubation/isolation, and pelletized by centrifugation. Enzymatic disaggregation may alter the relevant biological characteristics of adipose tissue, while providing release of complex, multiattachment of cell-to-cell and cell-to-matrix, effectively eliminating the bioactive ECM and periadventitial attachments. In many countries, the isolation of cellular elements is considered as a "more than minimal" manipulation, and is most often limited to controlled clinical trials and subject to regulatory review. Several alternative, nonenzymatic methods of adipose tissue processing have been developed to obtain via minimal mechanical manipulation an autologous tSVF product intended for delivery, reducing the procedure duration, lowering production costs, decreasing regulatory burden, and shortening the translation into the clinical setting. Ideally, these procedures might allow for the integration of harvesting and processing of adipose tissue for ease of injection, in a single procedure utilizing a nonexpanded cellular product at the point of care, while permitting intraoperative autologous cellular and tissue-based therapies. Here, we review and discuss the options, advantages, and limitations of the major strategies alternative to enzymatic processing currently developed for minimal manipulation of adipose tissue. Stem Cells Translational Medicine 2019;8:1265&1271.

Platelet-Rich Plasma and Micrografts Enriched with Autologous Human Follicle Mesenchymal Stem Cells Improve Hair Re-Growth in Androgenetic Alopecia. Biomolecular Pathway Analysis and Clinical Evaluation.

Platelet rich plasma (PRP) and Micrografts containing human follicle mesenchymal stem cells (HF-MSCs) were tried as a potential treatment for androgenetic alopecia (AGA). However, little to no work has yet to be seen wherein the bio-molecular pathway of HF-MSCs or PRP treatments were analyzed. The aims of this work are to report the clinical effectiveness of HF-MSCs and platelet-rich plasma evaluating and reviewing the most updated information related to the bio-molecular pathway. Twenty-one patients were treated with HF-MSCs injections and 57 patients were treated with A-PRP. The Wnt pathway and Platelet derived-growth factors effects were analyzed. 23 weeks after the last treatment with mean hair thickness increments (29 ± 5.0%) over baseline values for the targeted area. 12 weeks after the last injection with A-PRP mean hair count and hair density (31 ± 2%) increases significantly over baseline values. The increment of Wnt signaling in Dermal Papilla Cells evidently is one of the principal factors that enhances hair growth. Signaling from mesenchymal stem cells and platelet derived growth factors positively influences hair growth through cellular proliferation to prolong the anagen phase (FGF-7), inducing cell growth (ERK activation), stimulating hair follicle development (ß-catenin), and suppressing apoptotic cues (Bcl-2 release and Akt activation).

Fluid Cartilage as New Autologous Biomaterial in the Treatment of Minor Nose Defects: Clinical and Microscopic Difference Amongst Diced, Crushed, and Fluid Cartilage.

Developing cartilage constructs with injectability, appropriate matrix composition, and persistent cartilaginous phenotype remains an enduring challenge in cartilage repair. Fourteen patients with minor contour deformity were treated with fluid cartilage filler gently injected as autologous fluid graft in deep planes of defect of the nose that were close to the bone or the cartilage. A computerized tomographic scan control was performed after 12 months. Pearson's Chi-square test was used to investigate differences in cartilage density between native and newly formed cartilages. The endpoints were the possibility of using fluid cartilage as filler with aesthetic and functional improvement and versatility. Patients were followed up for two years. The constructs of fluid cartilage graft that were injected in the deep plane resulted in a persistent cartilage tissue with appropriate morphology, adequate central nutritional perfusion without central necrosis or ossification, and further augmented nasal dorsum without obvious contraction and deformation. This report demonstrated that fluid cartilage grafts are useful for cartilage regeneration in patients with outcomes of rhinoplasty, internal nasal valve collapse, and minor congenital nose aesthetics deformity.

Facial skin rejuvenation by autologous dermal microfat transfer in photoaged patients: Clinical evaluation and skin surface digital profilometry analysis.

Cumulative long-term exposure to solar ultraviolet radiation promotes premature skin aging characterized by wrinkle formation and reduced skin elasticity. In this study, we assessed whether microfat transfer could improve dermal and subcutaneous tissue thickness loss associated with photoaging. Twenty-one patients affected by facial photoaging (photodamage grade II-IV; age range 35-62 years; 19 females, 2 males; all of Caucasian origin) were treated using minimally invasive autologous dermal white fat transfer harvested with a recently designed microcannula. The results were determined by clinical assessment and patient self-evaluation and quantified by the Antera 3D® dermal digital device for noninvasive, objective, reliable, and accurate assessment of facial skin texture, color, and wrinkle characteristics. Compared with the pretreatment condition, the increment in soft tissue volume and improvement in skin quality and texture were assessed by a dermatologist after treatment. In addition, instrumental evaluation by digital skin profilometry of the treated areas revealed a 41% reduction in average wrinkle depth (7.29 ± 1.04 × 10-2 mm pretreatment vs. 4.31 ± 1.16 × 10-2 mm at 90 days posttreatment; p < 0.001), improved skin texture, more homogeneous and uniform skin color, and declined facial hemoglobin and melanin concentrations. The majority of patients (above 90%) reported improvements in self-perception. No significant complications were reported throughout the study. In conclusion, by using digital profilometry analysis as an objective and innovative tool to determine the outcome of treatment, we demonstrated that autologous microfat transfer is a safe and well-tolerated procedure with measurable beneficial effects on facial skin aging.

Blue honeysuckle fruit (Lonicera caerulea L.) from eastern Russia: phenolic composition, nutritional value and biological activities of its polar extracts.

In the present work we conducted a comprehensive chemical analysis of blue honeysuckle (Lonicera caerulea) spontaneously growing in eastern Russia. HPLC-DAD-ESI/MS analysis showed cyanidin-3-glucoside as the major constituent among phenolics, while nutritional analysis revealed fibre, protein, calcium and magnesium as the most important macro- and micronutrients, respectively. Fatty acid composition was dominated by polyunsaturated fatty acids, linoleic acid being the most abundant. Furthermore, we evaluated several in vitro biological activities such as antioxidant, antimicrobial, antiproliferative, wound healing and immunomodulatory effects of blue honeysuckle aqueous and ethanolic extracts that are often incorporated in food and nutraceutical preparations. While the fruit extracts were revealed to be potent radical scavengers with significant inhibition of ABTS radical, thus confirming the literature data, their inhibitory effects against microbial pathogens and tumor cell lines were negligible. The fruit aqueous extract did not show toxicity to human fibroblasts, but 24 h treatment with 150-200 µg per mL of extract slightly enhanced the cell migration when tested by scratched wound assay. Worth mentioning was the inhibitory effect displayed by the blue honeysuckle fruit aqueous extract on human lymphocytes.

New perspectives in the treatment of hidradenitis suppurativa: surgery and brewer's yeast-exclusion diet.

BACKGROUND: Hidradenitis suppurativa (HS), or acne inversa, is a chronic inflammatory skin disease characterized by abscesses, fistulating sinus tracts, and scarring. The exact pathogenesis of this disease remains unclear. We present our experience with HS and a possible cause of the disease and offer a surgical and clinical treatment that allows for disease resolution. METHODS: We studied 12 patients who presented with axillary and perineal fistulas and underwent surgical excision (repaired by skin grafts or healing by secondary intention) or localized treatments followed by diet modification. All patients were found to have a specific immunology IgG reaction to brewer's yeast and wheat and were put on a controlled brewer's yeast-free diet for 12 months and followed monthly. RESULTS: The diet demonstrated immediate stabilization of their clinical symptoms, and the skin lesions regressed over the 12-month treatment period. Similarly, all the patients demonstrated an immediate recurrence of skin lesions following accidental or voluntary consumption of beer or other foods containing brewer's yeast or wheat. The patients also demonstrated a return of their quality of life and activities, including the reestablishment of sexual activity in the patients with inguinal and perigenital lesions. CONCLUSION: The results of the investigation suggested that the patients had severe reactions to brewer's yeast. Surgery followed by the elimination of the foods containing or made with the yeast resulted in a rapid stabilization of the dermatologic manifestation and a slow, but complete, regression of the skin lesions within a year. Despite the small number of patients in our study, our study could provide the basis for extensive and multicentric studies in order to better investigate this disabling and rare disease.