Ilaria Baldelli

In vitro polydeoxyribonucleotide effects on human pre-adipocytes

Abstract.  Objectives: Adipose tissue is the most abundant and accessible source of adult stem cells. Human processed lipoaspirate contains pre‐adipocytes that possess one of the a characteristic pathways of multipotent adult stem cells and are able to differentiate in vitro into mesenchymal and also neurogenic lineages. Because stem cells have great potential for use in tissue repair and regeneration, it would be significant to be able to obtain large amounts of these cells in vitro. As demonstrated previously, purine nucleosides and nucleotides mixtures can act as mitogens for several cell types. The aim of this study was to evaluate the effects of polydeoxyribonucleotides (PDRN), at appropriate concentrations, on human pre‐adipocytes grown in a controlled medium, also using different passages, so as to investigate the relationship between the effect of this compound and cellular senescence, which is the phenomenon when normal diploid cells lose the ability to divide further. Materials and methods: Human pre‐adipocytes were obtained by liposuction. Cells from different culture passages (P6 and P16) were treated with PDRN at different experimental times. Cell number was evaluated for each sample by direct counting after trypan blue treatment. DNA assay and the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide test were also carried out in all cases. Results and Conclusions: PDRN seemed to promote proliferation of human pre‐adipocytes at both passages, but cell population growth increased in pre‐adipocyte at P16, after 9 days as compared to control. Our data suggest that PDRN could act as a pre‐adipocyte growth stimulator.

Superficial Enhanced Fluid Fat Injection (SEFFI) to Correct Volume Defects and Skin Aging of the Face and Periocular Region.

BACKGROUND Although recent research on micro fat has shown the potential advantages of superficial implantation and high stem cell content, clinical applications thus far have been limited. OBJECTIVES The authors report their experience with superficial enhanced fluid fat injection (SEFFI) for the correction of volume loss and skin aging of the face in general and in the periocular region. METHODS The finer SEFFI preparation (0.5 mL) was injected into the orbicularis in the periorbital and perioral areas, and the 0.8-mL preparation was injected subdermally elsewhere in the face. RESULTS The records of 98 consecutive patients were reviewed. Average follow-up time was 6 months, and average volume of implanted fat was 20 mL and 51.4 mL for the 0.5-mL and 0.8-mL preparations, respectively. Good or excellent results were achieved for volume restoration and skin improvement in all patients. Complications were minor and included an oil cyst in 3 patients. The smaller SEFFI quantity (0.5 mL) was well suited to correct volume loss in the eyelids, especially the deep upper sulcus and tear trough, whereas the larger SEFFI content was effective for larger volume deficits in other areas of the face, including the brow, temporal fossa, zygomatic-malar region, nasolabial folds, marionette lines, chin, and lips. CONCLUSIONS The fat administered by SEFFI is easily harvested via small side-port cannulae, yielding micro fat that is rich in viable adipocytes and stem cells. Both volumes of fat (0.5 mL and 0.8 mL) were effective for treating age-related lipoatrophy, reducing facial rhytids, and improving skin quality. LEVEL OF EVIDENCE 4 Therapeutic.

Skin Rejuvenation and Volume Enhancement with the Micro Superficial Enhanced Fluid Fat Injection (M-SEFFI) for Skin Aging of the Periocular and Perioral Regions.

Background Adipose-derived stromal and stem cells (ADSC) in autologous fat promises regenerative advantages, and injected into the dermal and subdermal layers, enhances rejuvenation and volume. However, extremely superficial fat injection with current techniques is limited. Objectives Efficacy and viability evaluation of fat harvested with extremely small side port (0.3 mm) cannulae without further tissue manipulation for the correction of aging/thin skin in the periocular and perioral regions. Methods Micro-superficial enhanced fluid fat injection (M-SEFFI) harvests adipose tissue with a multi-perforated cannula (0.3 mm), and autologous platelet rich plasma (PRP) is added. The tissue is injected into the dermal region of the periocular and perioral zones. Efficacy and viability were evaluated by histological and cell culture analysis. Clinical assessment included retrospective evaluation according to 1 = no effect, 2 = fair effect, 3 = good effect, 4 = excellent effect. Results Between June 2014 and July 2015, 65 patients (7 men; mean age 49.7 years) were treated with M-SEFFI. No intraoperative complications or visible lumpiness were recorded. Analysis demonstrated mature, viable adipocytes with a strong stromal component. Following PRP addition, there was a greater proliferation noted in the M-SEFFI compared to the SEFFI (0.5 mm). Mean follow-up was 4.1 months. Clinical assessment by surgeons and patients at 1 month was 3.52 and 3.74, and 6 months 3.06 and 2.6 respectively. Conclusions M-SEFFI is effective and viable for lump free skin rejuvenation and volume enhancement, through the extraction of smoother ADSC rich, autologous fat tissue that does not require further tissue manipulation, to correct skin aging. Level of Evidence 4 Therapeutic

Interaction Between Breast Cancer Cells and Adipose Tissue Cells Derived from Fat Grafting

BACKGROUND Adipose tissue transplantation has the benefit of providing both regenerative and aesthetic outcomes in breast cancer treatment. However, the transplanted tissue can stimulate the growth of residual cancer cells. OBJECTIVES The aim of this study is to identify the interactions between adipose tissue cell subpopulations and human cancer cell lines. METHODS Intact adipose tissue from lipofilling procedures as well as fibroblasts derived from adipose tissue, were cocultured in the presence of MDA-MB-231, MCF-7 e ZR-75-1 breast cancer cell lines. The influence on cancer cell lines of fibroblasts, induced to differentiate into specific adipocytes, was also assayed. RESULTS All cancer cell lines displayed a significant increase in proliferation rate when cocultured in the presence of either intact adipose tissue or induced adipocytes. To a lesser extent, uninduced fibroblasts stimulate breast cancer cell proliferation. CONCLUSIONS Recent studies have shown that the microenvironment surrounding breast cancer cells may stimulate growth and promote progression of residual cancer cells when surgery is performed on the main tumor mass. Accordingly, the graft of adipose tissue could potentially promote or accelerate the development of a subclinical tumor or support its locoregional recurrence. Our data suggest that adipocytes have a remarkable influence on the proliferation of cancer cell lines. The oncological safety of the lipofilling procedure outcome is still debated; thus, further studies and consistent follow-up examination are needed.

Implant-based Breast Reconstruction Using a Polyester Mesh (surgimesh-pet): A Retrospective Single-center Study

Background: Although the introduction of acellular dermal matrices is considered one of the most important advancements in alloplastic breast reconstruction, costs and local policy limit their use in Italy. The purpose of this study was to assess short-term outcomes following Surgimesh-PET–based breast reconstruction. Methods: A single-center, retrospective, case-control study was performed from January 1, 2012, to December 31, 2013, by enrolling 206 breast reconstructions performed in 196 patients after oncologic (n = 200) or prophylactic (n = 6) mastectomy. Group A included 63 patients who underwent 70 immediate Surgimesh-PET–assisted breast reconstructions, and group B included 133 patients who underwent 136 standard breast reconstructions. Results: No significant differences between groups occurred for early postoperative complications (p = 0.610), major complications that required surgical revision (p = 0.887), volume (p = 0.498) or width of the prosthesis (p = 0.201), skin-sparing mastectomy (p = 0.315), or axillary surgery (p = 0.265). Multivariate logistic regression showed that prior radiotherapy was the only significant variable for early postoperative complications in both whole series (p = 0.011) and group B (p = 0.046), whereas body mass index greater than 25 was an independent predictor in group A (p = 0.041). Prior radiotherapy was the only variable that reached statistical significance in the multivariate model for major complications in the whole series (p = 0.005). Conclusions: Short-term outcomes of Surgimesh-PET–based alloplastic breast reconstruction are promising. Further studies are needed to evaluate the long-term results of this surgical approach. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, III.

Phenotypic and genotypic characterization of Staphylococci causing breast peri-implant infections in oncologic patients

BackgroundStaphylococcus epidermidis and S. aureus have been identified as the most common bacteria responsible for sub-clinical and overt breast implant infections and their ability to form biofilm on the implant as been reported as the essential factor in the development of this type of infections. Biofilm formation is a complex process with the participation of several distinct molecules, whose relative importance in different clinical settings has not yet been fully elucidated. To our knowledge this is the first study aimed at characterizing isolates causing breast peri-implant infections.ResultsThirteen S. aureus and seven S. epidermidis causing breast peri-implant infections were studied.Using the broth microdilution method and the E-test, the majority of the strains were susceptible to all antibiotics tested. Methicillin resistance was detected in two S. epidermidis. All strains had different RAPD profiles and were able to produce biofilms in microtitre plate assays but, while all S. aureus carried and were able to express icaA and icaD genes, this was only true for one S. epidermidis. Biofilm development was glucose- and NaCl-induced (5 S. aureus and 1 S. epidermidis) or glucose-induced (the remaining strains). Proteinase K and sodium metaperiodate treatment had different effects on biofilms dispersion revealing that the strains studied were able to produce chemically different types of extracellular matrix mediating biofilm formation.All S. aureus strains harboured and expressed the atlA, clfA, FnA, eno and cna genes and the majority also carried and expressed the sasG (10/13), ebpS (10/13) genes.All S. epidermidis strains harboured and expressed the atlE, aae, embp genes, and the majority (six strains) also carried and expressed the fbe, aap genes.Genes for S. aureus capsular types 5 and 8 were almost equally distributed. The only leukotoxin genes detected were lukE/lukD (6/13).ConclusionsS. aureus and S. epidermidis breast peri-implant infections are caused by heterogeneous strains with different biofilm development mechanisms.Since the collagen adhesin (cna) gene is not ubiquitously distributed among S. aureus, this protein could have an important role in the cause of breast peri-implant infections.

Conscientious Objection to Animal Experimentation in Italian Universities

Simple Summary This paper examines the trend of Italian academic faculties in complying with the obligation to inform university students of their right to exercise their conscientious objection to scientific or educational activities involving animals, hereafter written as “animal CO”, as established by Law 413/1993, “Norme sull’obiezione di coscienza alla sperimentazione animale” (“Rules on conscientious objection to animal experimentation”), thereafter “Law 413/1993”. Despite an increasing interest in the principles of animal ethics by the international community, this law is still largely disregarded more than 20 years after its enactment. The Ethics Committees, Animal Welfare Committees, as well as the Italian Ministry of Education, University and Research should preside over and monitor the Universities’ compliance with the duty to disclose animal CO. Abstract In Italy, Law 413/1993 states that public and private Italian Institutions, including academic faculties, are obliged to fully inform workers and students about their right to conscientious objection to scientific or educational activities involving animals, hereafter written as “animal CO”. However, little monitoring on the faculties’ compliance with this law has been performed either by the government or other institutional bodies. Based on this premise, the authors have critically reviewed the existing data and compared them with those emerging from their own investigation to discuss limitations and inconsistencies. The results of this investigation revealed that less than half of Italian academic faculties comply with their duty to inform on animal CO. Non-compliance may substantially affect the right of students to make ethical choices in the field of animal ethics and undermines the fundamental right to express their own freedom of thought. The Italian Ministry of Education, Universities and Research, ethics committees and animal welfare bodies should cooperate to make faculties respect this law. Further research is needed to better understand the reasons for the current trend, as well as to promote the enforcement of Law 413/1993 with particular regard to information on animal CO.

Aesthetic Evaluation in Oncoplastic and Conservative Breast Surgery: A Comparative Analysis

Background: In conservative breast surgery, the achievement of a satisfactory cosmetic result could be challenging; oncoplastic techniques may be helpful in many cases. A comparative analysis was performed among 3 groups of patients undergoing oncoplastic techniques plus external radiation therapy or intraoperative radiotherapy (IORT) and breast conservative surgery plus external radiation therapy; long-term oncologic results in terms of disease relapse and aesthetic outcomes were compared. Methods: Ninety-six patients were considered: 32 patients treated with oncoplastic surgery, 16 then subjected to radiotherapy (group 1) and another 16 treated with IORT (group 2); 64 patients treated by conservative surgery and radiotherapy formed the control group (group 3). Patients were asked to give a judgment on the cosmetic result considering the following parameters: breast symmetry, appearance of the residual scar, symmetry between the 2 nipple-areola complexes, global aesthetic judgment, and satisfaction about the result. Results: With respect to the oncological and aesthetic outcome, the statistical significance of the results obtained in the 3 groups was calculated using the chi-square test. The results, processed by the chi-square test, were not statistically significant; however, the overall judgments expressed by the patients of all 3 groups were more than satisfactory (scores greater than or equal to 6). Conclusions: In our experience, when the inclusion criteria are satisfied and the equipment is available, oncoplastic techniques associated with IORT should be considered the treatment of choice for breast cancer in early stage. The excellent cosmetic results and patient’s satisfaction encourage us to continue on this way.

A Survey on the Knowledge and Attitudes of Italian Medical Students toward Body Donation: Ethical and Scientific Considerations

Post mortem body donation (PMBD) for medical training and research plays a key role in medical-surgical education. The aim of this study is to evaluate Italian medical students’ awareness and attitudes regarding this practice. A questionnaire was sent to 1781 Italian medical students (MS). A total of 472 MS responded: 406 (92.91%) had a strongly positive attitude to PMBD, while 31 (7.09%) were not in favor. The majority of subjects were Catholic (56.36%), while 185 and 21 subjects, said that they did not hold any religious beliefs, or were of other religions, respectively. Multivariate analysis showed significant associations (p-values < 0.05) between PMBD and religion, as well as perceptions of PMBD as an act of altruism, a tool for learning surgical practices, body mutilation, and an act contrary to faith. Although Italian MS believed cadaver dissection to be an important part of their education, they did not know much about it and had not received training on this altruistic choice. As future doctors, MS can play an important role in raising public awareness of the importance of PMBD for medical education and research. Specific educational programs to improve knowledge of this topic among MS are needed.

Analysis of clinical management of infected breast implants and of factors associated to successful breast pocket salvage in infections occurring after breast reconstruction.

OBJECTIVES Considerable efforts have been devoted so far to improve salvage procedures of infected breast implants in absence of defined guidelines or validated clinical protocols. Within a cohort of prospectively recruited patients who underwent breast reconstruction, we performed a retrospective review of proven implant infections in order to describe factors contributing to management success. METHODS We collected data in 1293 consecutive patients who underwent two stage (expander+prosthesis) breast reconstruction with at least 12 months of follow-up. Demographic data, timing of infection, type of microorganism, intent of salvage, fate of the implant, type of antibiotic treatment and follow-up were recorded in a prospective data collection on clinical records. RESULTS Implant infections occurred in 103 of 1293 patients (8%). Among these, 73 (71%) were proven infections with confirmed microbiology. Implant pocket salvage was attempted in 43/73 (59%). patients A higher proportion of expander implant pockets were successfully saved compared to prosthetic pocket (p=0,04). Gram-positive microrganisms represented the majority of etiologic agents, with coagulase negative staphylococci prevailing over Staphylococcus aureus. No association was observed between success rate and type of infecting microorganism. A higher proportion of patients with previous or intraoperative radiotherapy or with perioperative chemotherapy underwent an attempt of implant salvage (p=0,081 and 0,0571 trend, respectively). No single antibiotic regimen was superior to the others in terms of success rate. Implant pocket salvage was higher in expanders compared to prostheses (74% vs 33% p=0,04). Higher success rates in implant pocket salvage were evident when implant replacement was preceded and followed by antibiotic treatment compared to inpatient antibiotic treatment alone (100% versus 57%, p=0,035). CONCLUSION Patient selection in clinical practice leads to differences in patients with breast implant infection who are considered for attempts at implant salvage vs. those who are treated with implant removal. Salvage of breast implant pockets can be obtained in the majority of patients with combined one-step implant replacement surgery and antibiotic treatment. Increased efforts and protocols to recruit patients into pocket salvage management are needed.