Neural differentiation of human adipose-derived mesenchymal stem cells induced by glial cell conditioned media.

Adipose-derived mesenchymal stem cells (ASCs) may transdifferentiate into cells belonging to mesodermal, endodermal, and ectodermal lineages. The aim of this study was to verify whether a neural differentiation of ASCs could be induced by a conditioned medium (CM) obtained from cultures of olfactory ensheathing cells (OECs) or Schwann cells (SCs). ASCs were isolated from the stromal vascular fraction of adipose tissue and expanded for 2-3 passages. They were then cultured in OEC-CM or SC-CM for 24 hr or 7 days. At each stage, the cells were tested by immunocytochemistry and flow cytometer analysis to evaluate the expression of typical neural markers such as Nestin, PGP 9.5, MAP2, Synapsin I, and GFAP. Results show that both conditioned media induced similar positive effects, as all tested markers were overexpressed, especially at day 7. Overall, an evident trend toward neuronal or glial differentiation was not clearly detectable in many cases. Nevertheless, a higher tendency toward a neuronal phenotype was recognized for OEC-CM (considering MAP2 increases). On the other hand, SC-CM would be responsible for a more marked glial induction (considering GFAP increases). These findings confirm that environmental features can induce ASCs toward a neural differentiation, either as neuronal or glial elements. Rather than supplementing the culture medium by adding chemical agents, a "more physiological" condition was obtained here by means of soluble factors (cytokines/growth factors) likely released by glial cells. This culture strategy may provide valuable information in the development of cell-based therapeutic approaches for pathologies affecting the central/peripheral nervous system.

Adipose-derived stem cells: Comparison between two methods of isolation for clinical applications.

BACKGROUND: Adipose-derived stem cells are recognized as being an effective mesenchymal stem cell population with enormous potential in different fields of regenerative medicine and stem cell therapy. Although there is unanimous agreement on the harvesting procedure for adipose tissue, there are various protocols for adipose-derived stem cell isolation. The aim of this study was compare two methods of adipose-derived stem cells (ASCs) isolation, one based on a mechanical + enzymatic (ME) procedure and the other one exclusively mechanical (MC), in order to determine which one was superior to the other in accordance with current European and US legislation. METHODS: We reported step by step the two different methods ASCs isolation by comparing them. The ME procedure included the use of a centrifuge, an incubator and collagenase digestion solution (Collagenase NB 6 GMP Grade 17458; Serva GmbH, Heidelberg, Germany). The MC procedure was performed by vibrating shaker and centrifuge, both placed in a laminar airflow bench. RESULTS: With the ME procedure, a mean of 9.06 × 105 ASCs (range, 8.4 to 9.72 × 105; SD ± 6.6 × 105) was collected, corresponding to 25.9% of the total number of harvested cells. With the MC procedure, a mean of 5 × 105 ASCs (range: 4.0 to 6.0 × 105; SD, ±1 × 105) was collected, corresponding to 5% of the total number of harvested cells. CONCLUSION: Based on data collected, from the same amount of lipoaspirate the ME procedure allowed to isolate a greater number of ASCs (25,9%) compared to the MC one (5%).

Necrotizing Fasciitis in Aesthetic Surgery: A Review of the Literature.

INTRODUCTION: Necrotizing fasciitis (NF) is a rare, potentially fatal, infective complication that can occur after surgery. Diagnosis is still difficult and mainly based on clinical data. Only a prompt pharmacological and surgical therapy can avoid dramatic consequences. There are few reports regarding NF as a complication after aesthetic surgical procedures, and a systematic review still lacks. MATERIALS AND METHODS: We have performed a systematic review of English literature on PubMed, covering a period of 30 years. Keywords used were "necrotising fasciitis" matched with "aesthetic surgery complications", "breast surgery", "mammoplasty", "blepharoplasty", "liposuction", "facelift", "rhinoplasty fasciitis", "arm lift", "thigh lift", "otoplasty" and "abdominoplasty fasciitis". No additional search and temporal limitation were set. RESULTS: Among 3782 papers concerning NF, only 18 were related to NF after an aesthetic surgical procedure. Liposuction was the most affected procedure, with buttocks and lower extremity the most involved anatomical regions. The majority of the infections were monomicrobial, promoted by Streptococcus pyogenes. In most cases, NF occurred within the third post-operative day with non-specific signs and symptoms. In 14 cases, a single or multiple surgical interventions were performed and survival was achieved in 11 patients. CONCLUSIONS: In case of infection after aesthetic surgery, we should always bear in mind NF. Clinical hallmarks still guide NF management. Because early signs and symptoms are usually non-specific, a strict clinical control is highly suggested. Once clinical suspicion is raised, prompt antibacterial therapy should be administered, followed by surgical debridement in case of ineffective response. LEVEL OF EVIDENCE III: 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 .

Lubricin in synovial fluid of mild and severe temporomandibular joint internal derangements

BACKGROUND: To understand the molecular basis of temporomandibular joint (TMJ) pathologies, we aimed to investigate the lubricin levels in the TMJ synovial fluid (SF) of patients with mild to severe internal derangements (IDs). MATERIAL AND METHODS: A total, 34 joints were the study group. Only patients, with a Wilkes stage of III, IV and V were included, in this sample. Control group consisted of SF from eight joints, from patients undergoing to orthognatic surgery. Concentrations of lubricin in the SF from both samples were measured using ELISA system. RESULTS: The mean lubricin concentration was 7.029 ± 0.21 mig/mL in stage III patients; 5.64 ± 0.10 mig/mL in stage IV patients, and 4.78 ± 0.11 mig/mL in stage V patients. The lubricin levels from stage IV and stage V patients differed significantly (P ≤ 0.001) from those of control subjects. Lubricin levels were inversely correlated with age and to VAS score. CONCLUSIONS: The results of this cross-sectional study highlight the relationship between disease severity and the levels of lubricin in TMJ SF. Our findings suggest that novel biotherapeutic approaches, including the administration of recombinant lubricin in the joint cavity, for the treatment of TMJ diseases can be developed

Vulvar Melanoma Associated with Lichen Sclerosus in a Child: Case Report and Literature Review.

The authors describe the sixth pediatric case to date of primary vulvar melanoma associated with lichen sclerosus and propose a practical management for such a rare cancer.

Reconstruction of a Nose Bitten and Eaten by a Dog: A Ten-Year Follow-up of a Bitten-Off Nose Replantation Performed on an Eleven-Year-Old Boy.

The nose is one of the most important aesthetic units of the face. After a traumatic amputation, replantation is undoubtedly the procedure of choice, although, technically, very challenging. We report the 10-year follow-up of a partially digested nose replantation after a dog-bite in an 11-year-old boy using supermicrosurgery technique. Our report confirms that the microsurgical replantation of the nose can lead to an acceptable aesthetic result, with the sufficient growth of the replanted part in the following years. Besides the patient did not report sensibility disorders. The nose replantation in our opinion is the best reconstructive option to achieve an optimal aesthetic and functional outcome.

Coverage of the neurovascular unit of the fingertip using a reverse homodigital dorsal flap.

The exposure of bone, tendons, vessels, and nerves in a digital defect is one of the most frequent and severe problems to solve in hand surgery and current approaches are still disappointing. We show the use of an homodigital adipofascial flap taken from the same finger for covering the pulpar defect in a one-step surgical technique able to preserve the digital artery.

Decrease of apoptosis markers during adipogenic differentiation of mesenchymal stem cells from human adipose tissue.

Although the proliferation and differentiation of mesenchymal stem cells (MSCs) from adipose tissue (AT) have been widely studied, relatively little information is available on the underlying mechanism of apoptosis during the adipogenic differentiation. Thus, the aim of this study was to analyze how the expression of some apoptotic markers is affected by in vitro expansion during adipogenic differentiation of AT-MSCs. The cultures incubated or not with adipogenic medium were investigated by Western blot at 7, 14, 21, and 28 days for the production of p53, AKT, pAKT, Bax, PDCD4 and PTEN. MSCs were recognized for their immunoreactivity to MSC-specific cell types markers by immunocytochemical procedure. The effectiveness of adipogenic differentiation was assessed by staining with Sudan III and examination of adipogenic markers expression, such as PPAR-γ and FABP, at different time points by Western blot. The adipogenic differentiation medium led to the appearance, after 7 days, of larger rounded cells presenting numerous vacuoles containing lipids in which it was evident a red-orange staining, that increased in size in a time-dependent manner, parallel to an increase of the levels of expression of PPAR-γ and FABP. More than 50 % of human MSCs were fully differentiated into adipocytes within the four-week induction period. The results showed that during adipogenic differentiation of AT-MSCs the PI3K/AKT signaling pathway is activated and that p53, PTEN, PDCD4, and Bax proteins are down-regulated in time-dependent manner. Our data provide new information on the behavior of some apoptotic markers during adipogenic differentiation of AT-MSCs to apply for tissues repair and regeneration.