Mario Cherubino

Adipose-derived stem cells for wound healing applications.

Nonhealing wounds remain a significant challenge for plastic surgeons. More than 600,000 people suffer from venous ulcers and 1.5 to 3 million people are being treated for pressure sores every year in the United States. The use of tissue engineering techniques such as stem-cell therapy and gene therapy to improve wound healing is a promising strategy. Adipose tissue represents a source of cells that may be able to enhance wound healing. Adipose-derived stem cells (ASCs) are adult stem cells that are easily harvested and of great interest for plastic surgeons. Specifically, ASCs secrete angiogenic growth factors that can induce tissue regeneration. This review describes innovative research strategies using ASCs therapies for treatment of chronic, nonhealing wounds.

Adipose-derived stem cells for soft tissue reconstruction

In soft tissue repair, there are several surgical options such as nondegradable, inert, synthetic, biodegradable implants or autologous tissue transplantation. However, the potential of using autologous adult stem cells derived from fat tissue is quickly becoming a clinical reality. The possibility of using an abundant source of extraneous tissue as a soft tissue implant has significant implications for plastic and reconstructive surgeons. This strategy would be particularly useful after tumor removal or trauma. The ability of adult stem cells derived from adipose tissue (termed adipose-derived stem cell) to proliferate and differentiate in vivo or in vitro is actively being studied owing to the potential implementation in reconstructive surgery. This review describes innovative research strategies and discusses the first clinical studies involving adipose-derived stem cells as a motif for soft tissue reconstruction.

Cosmetic outcomes of various skin closure methods following cesarean delivery: a randomized trial

OBJECTIVEnThe objective of the study was to compare scar quality associated with different types of wound closure methods after cesarean section (CS).nnnSTUDY DESIGNnPatients were randomized to have skin closure following CS with either staples or 3 different types of subcuticular sutures. Scar quality was evaluated 2 and 6 months postoperatively. The Vancouver Scar Scale, the Patient and Observer Scar Assessment Scale (POSAS), and a visual analog scale were used as scar assessment tools.nnnRESULTSnOf the 180 patients who were recruited, 123 successfully completed the study. No difference in both subjective and objective scar rating was detected across groups at either 2 months or 6 months. In the overall study population, objective scores correlated with patient rating, and correlation was strongest between the observer and patient components of the POSAS (r = 0.48).nnnCONCLUSIONnIn women undergoing CS, stapled wounds and those closed with subcuticular sutures result in equivalent cosmetic appearance of the scar.

Masquelet technique for the treatment of a severe acute tibial bone loss

The induced membrane technique described by Masquelet has been used successfully for many years for posttraumatic bone defect reconstruction, non-unions and osteomyelitis. The main advantages are the two-step surgical procedure that in case of primary infection allows repeated debridement if necessary, in case of internal fixation early weight bearing with decreased malalignment risk and it has a short learning curve. A theoretical application of this procedure is the management of acute severe traumatic bone loss of the limbs despite the lack of this experience in literature. We report on a Gustilo IIIB meta-epiphyseal fracture (AO 43-C3) of the leg with a 6 cm in length bone loss that was treated with the Masquelet technique.

Acellular dermal matrices and radiotherapy in breast reconstruction: a systematic review and meta-analysis of the literature.

The increasing use of commercially available acellular dermis matrices for postmastectomy breast reconstruction seems to have simplified the surgical procedure and enhanced the outcome. These materials, generally considered to be highly safe or with only minor contraindications due to the necessary manipulation in preparatory phases, allow an easier one-phase surgical procedure, in comparison with autologous flaps, offering a high patient satisfaction. Unfortunately, the claim for a higher rate of complications associated with irradiation at the implant site, especially when the radiation therapy was given before the reconstructive surgery, suggested a careful behaviour when this technique is preferred. However, this hypothesis was never submitted to a crucial test, and data supporting it are often discordant or incomplete. To provide a comprehensive analysis of the field, we searched and systematically reviewed papers published after year 2005 and registered clinical trials. On the basis of a meta-analysis of data, we conclude that the negative effect of the radiotherapy on the breast reconstruction seems to be evident even in the case of acellular dermis matrices aided surgery. However, more trials are needed to make solid conclusions and clarify the poor comprehension of all the factors negatively influencing outcome.

Human adipose-derived stem cells promote vascularization of collagen-based scaffolds transplanted into nude mice

Aim: After in vivo implantation of cell-loaded devices, only the cells close to the capillaries can obtain nutrients to maintain their functions. It is known that factors secreted by stem cells, rather than stem cells themselves, are fundamental to guarantee new vascularization in the area of implant. Materials & methods: To investigate this possibility, we have grafted mice with Bilayer and Flowable Integra® scaffolds, loaded or not with human adipose-derived stem cells. Results: Our results support the therapeutic potential of human adipose-derived stem cells to induce new vascular networks of engineered organs and tissues. Conclusion: This finding suggests that our approach can help to form new vascular networks that allow sufficient vascularization of engineered organs and tissues in cases of difficult wound healing due to ischemic conditions.

Treatment of primary axillary hyperhidrosis with botulinum toxin type a: our experience in 50 patients from 2007 to 2010.

Background. Local injections of Botulinum toxin type A (BTX-A) are an effective and safe solution for primary bilateral axillary hyperhidrosis. Traditional treatments are often ineffective and difficult to tolerate. This study was performed to assess the efficacy and safety of Botulinum toxin type A in the treatment of these diseases and to evaluate the reliability of patients subjective rating in the timing of repeat injections. Methods. From 2007 to 2008, we included in the study and treated a total of 50 patients, and we used the Minors iodine test and the hyperhidrosis diseases severity scale as initial inclusion criteria and also for evaluating the followup, comparing to patients subjective rating. We used also a specific questionnaire to evaluate the level of pain, the onset of the effect, any eventual adverse effect of the treatment, the onset of compensatory hyperhidrosis, and the global grade of satisfaction. The data were analyzed using standard statistical methods. Results. 88% of patients were totally satisfied and all patients repeated the treatment during all the study. The symptom-free interval was in median 6 months with an average improving of HDSS of 1.5 points. In 86%, there was a complete accordance between the subjective patients demand of the repetition of the treatment and the positivity to Minor test and HDSS. No major side effects happened. Conclusion. Local injections of Botulinum toxin type A (BTX-A) result in an effective and safe solution for bilateral axillary primary hyperhidrosis for the absence of significant morbidity, side effects, and lack of efficacy or duration. The only defects are the need of repetition of the treatment and relative costs.

Osteochondritis dissecans of the entire femoral trochlea.

Juvenile osteochondritis dissecans (JOCD) is a term used to describe the separation of an articular cartilage and subchondral bone segment from the remaining articular surface in skeletally immature children. Maximum incidence is between the ages of 10 and 20 years. It is found more frequently in children who are athletes and is twice as common in boys as it is in girls. This condition most commonly affects the knee joint, with approximately 85% of lesions occurring in the medial femoral condyle and 15% of lesions occurring in the lateral femoral condyle. There are very few cases reported in the literature that involve the patellofemoral joint. The patella is more frequently affected, whereas osteochondritis dissecans of the trochlea femuri was first described by Axhausen in 1912. Review of the literature reveals few reported JOCD cases of either the lateral or the medial femoral trochlear groove. We report a rare case of JOCD involving the entire femoral trochlea and its management.

Identification of reference genes for qPCR analysis during hASC long culture maintenance

Up to now quantitative PCR based assay is the most common method for characterizing or confirming gene expression patterns and comparing mRNA levels in different sample populations. Since this technique is relative easy and low cost compared to other methods of characterization, e.g. flow cytometry, we used it to typify human adipose-derived stem cells (hASCs). hASCs possess several characteristics that make them attractive for scientific research and clinical applications. Accurate normalization of gene expression relies on good selection of reference genes and the best way to choose them appropriately is to follow the common rule of the “Best 3”, at least three reference genes, three different validation software and three sample replicates. Analysis was performed on hASCs cultivated until the eleventh cell confluence using twelve candidate reference genes, initially selected from literature, whose stability was evaluated by the algorithms NormFinder, BestKeeper, RefFinder and IdealRef, a home-made version of GeNorm. The best gene panel (RPL13A, RPS18, GAPDH, B2M, PPIA and ACTB), determined in one patient by IdealRef calculation, was then investigated in other four donors. Although patients demonstrated a certain gene expression variability, we can assert that ACTB is the most unreliable gene whereas ribosomal proteins (RPL13A and RPS18) show minor inconstancy in their mRNA expression. This work underlines the importance of validating reference genes before conducting each experiment and proposes a free software as alternative to those existing.

A simple and reliable method to perform biomechanical evaluation of postoperative nerve adhesions.

BACKGROUNDnPerineural fibrotic adhesions are among the major complications of peripheral nerve surgery. While different experimental models have been used for the pre-clinical testing of anti-adherential strategies, the methods used so far to induce scar tissue appear to be poorly standardized and reproducible.nnnNEW METHODnThirty adult mice were used. Two methods were tested: the first one is based on burning the perineural muscular bed with a diathermocoagulator, while the second is based on direct scratching of the nerve surface with a cotton swab. After 3 weeks, the fibrotic reaction was assessed by measuring the peak pull out force of the nerve from muscular bed by means of a new tool specifically devised for biomechanical assessment of scar tissue formation. Moreover, histological analysis with specific collagen stain was also carried out.nnnRESULTSnBoth methods produced fibrotic reaction. Statistical analysis of biomechanical data showed a significant difference between burning and scratching group compared to the control sham operated group. No significant differences were detected between burning and scratching group. Histological analysis showed the presence of perineural scar tissue in both groups, though with a different distribution pattern.nnnCOMPARISON WITH OTHER METHODSnThis protocol is easier to perform. The tool used for biomechanical evaluation is reliable and cheap.nnnCONCLUSIONSnBoth methods for perineural scar formation are effective and simple. They represent reproducible models for the study of the anti-adherential strategies. Yet, biomechanical testing with the device that we have developed proved to be a reliable and simple method for the quantitative assessment of the degree of perineural adhesion formation.