3D co-cultures of osteoblasts and endothelial cells in DegraPol foam: Histological and high field MRI analyses of pre-engineered capillary networks in bone grafts

Tissue engineering of bone grafts was addressed in a critical size model on the chick chorioallantoic membrane model (CAM assay), using DegraPol(R) (DP) foam as scaffold material. The scaffolds were seeded with cultures of human osteoblasts (OB) and human en notdo notthelial cells (EC), respectively, or with a co-culture of the two cell types (control: no cells). In vitro samples (7 days cultivation) and ex vivo CAM samples at incubation day 15 (ID 15) were analyzed by high field magnetic resonance imaging (MRI) and histology. The co-culture system performed best with respect to perfusion, as assessed by contrast-enhanced MRI using Gd-DTPA. The scaffold seeded by the co-culture supported an increased vascular ingrowth, which was confirmed by histological analysis. DP foam is a suitable scaffold for bone tissue engineering and the MRI technique allows for non-destructive and quantitative assessment of perfusion capability during early stages of bone forming constructs

Thermal conditioning improves quality and speed of keratinocyte sheet production for burn wound treatment

BACKGROUND AIMS Cultured patient-specific keratinocyte sheets have been used clinically since the 1970s for the treatment of large severe burns. However, despite significant developments in recent years, successful and sustainable treatment is still a challenge. Reliable, high-quality grafts with faster availability and a flexible time window for transplantation are required to improve clinical outcomes. METHODS Keratinocytes are usually grown in vitro at 37°C. Given the large temperature differences in native skin tissue, the aim of the authors' study was to investigate thermal conditioning of keratinocyte sheet production. Therefore, the influence of 31°C, 33°C and 37°C on cell expansion and differentiation in terms of proliferation and sheet formation efficacy was investigated. In addition, the thermal effect on the biological status and thus the quality of the graft was assessed on the basis of the release of wound healing-related biofactors in various stages of graft development. RESULTS The authors demonstrated that temperature is a decisive factor in the production of human keratinocyte sheets. By using specific temperature ranges, the authors have succeeded in optimizing the individual manufacturing steps. During the cell expansion phase, cultivation at 37°C was most effective. After 6 days of culture at 37°C, three times and six times higher numbers of viable cells were obtained compared with 33°C and 31°C. During the cell differentiation and sheet formation phase, however, the cells benefited from a mildly hypothermic temperature of 33°C. Keratinocytes showed increased differentiation potential and formed better epidermal structures, which led to faster biomechanical sheet stability at day 18. In addition, a cultivation temperature of 33°C resulted in a longer lasting and higher secretion of the investigated immunomodulatory, anti-inflammatory, angiogenic and pro-inflammatory biofactors. CONCLUSIONS These results show that by using specific temperature ranges, it is possible to accelerate the large-scale production of cultivated keratinocyte sheets while at the same time improving quality. Cultivated keratinocyte sheets are available as early as 18 days post-biopsy and at any time for 7 days thereafter, which increases the flexibility of the process for surgeons and patients alike. These findings will help to provide better clinical outcomes, with an increased take rate in severe burn patients

Delayed enzymatic debridement in severe burns: Proof of concept

Introduction Enzymatic debridement (ED) is a novel powerful therapy for debridement of severe burns. Standard ED is usually performed within 72 h after injury following a presoaking phase. Little evidence exists on the effectiveness of ED later than 72 h after trauma. In this retrospective study, we compared outcomes of burn patients treated within versus later than 72 h after injury. Patients and Methods 110 patients with severe burns treated with ED between 2016 and 2020 were evaluated. Patients treated later than 72 h after trauma were identified and matched to a control group treated within 72 h. Matching criteria included age, area treated with ED, and localization of ED. Exclusion criteria were abbreviated burn severity index (ABSI) greater than 12 and death within the first 10 days after burn injury. Primary outcomes were time to full epithelialization and number of secondary surgical interventions. Results 16 patients (11 female, 5 male) matched the inclusion criteria and were assigned to the late treatment group. Mean age was 54.0 ± 19.0 years, the = and mean ABSI score 6.3 ± 3.2. 16 matched patients were assigned to the early ED group. Secondary surgical procedures were performed in 62.5% of cases in both groups with a mean of 1.7 (late treatment) vs. 2.2 (control; p = 0.29) secondary procedures in each group, respectively. No significant difference between groups regarding time to complete epithelialization (28.2 days vs. 27.3 days, p = 0.45) was observed. Infection rate was higher (18.8% vs. 6.3%, p = 0.28) in the delayed group. Conclusion Delayed ED is a feasible procedure as part of personalized care in burn surgery. In our retrospective study, we could not identify r safety issues except a slightly higher infection rate. This may however be attributed to delayed initiation of burn treatment itself

Sensitization and desensitization of burn patients as potential candidates for vascularized composite allotransplantation

Sensitization describes the acquired ability of the immune system to react to foreign human leukocyte antigens (HLA) by producing antibodies and developing memory cells. In the field of transplantation, recipient preformed HLA antibodies due to previous sensitization have been identified - beneath ABO incompatibility - as a major factor for acute graft rejection. Several reasons for sensitization have largely been studied, such as previous blood transfusions, pregnancies or former transplants. Recent studies indicate that the use of assist devices (e.g. ECMO) or cadaveric skin allotransplantation providing temporary coverage in burn patients may lead to additional sensitization. As vascularized composite allotransplantation (VCA) has become a rapidly advancing therapeutic option for reconstruction of complex tissue defects in burns, it seems even more important to become familiar with immunological principles and to be cautiously aware of both sources of sensitization and therapeutic concepts in burns avoiding sensitization. This may also include emergency VCAs in burn patients as potential strategy for early definitive reconstruction avoiding procedures triggering HLA antibody formation. We hereby provide an overview on current evidence in the field of pre- and peritrans-plant sensitization, followed by posttransplant strategies of desensitization and their potential impact on future treatments of burn patients. (C) 2015 Elsevier Ltd and ISBI. All rights reserved.Peer reviewe

Increased levels of VEGF-C and macrophage infiltration in lipedema patients without changes in lymphatic vascular morphology

Lipedema is a chronic adipose tissue disorder characterized by the disproportional subcutaneous deposition of fat and is commonly misdiagnosed as lymphedema or obesity. The molecular determinants of the lipedema remain largely unknown and only speculations exist regarding the lymphatic system involvement. The aim of the present study is to characterize the lymphatic vascular involvement in established lipedema. The histological and molecular characterization was conducted on anatomically-matched skin and fat biopsies as well as serum samples from eleven lipedema and ten BMI-matched healthy patients. Increased systemic levels of vascular endothelial growth factor (VEGF)-C (P=0.02) were identified in the serum of lipedema patients. Surprisingly, despite the increased VEGF-C levels no morphological changes of the lymphatic vessels were observed. Importantly, expression analysis of lymphatic and blood vessel-related genes revealed a marked downregulation of Tie2 (P<0.0001) and FLT4 (VEGFR-3) (P=0.02) consistent with an increased macrophage infiltration (P=0.009), without changes in the expression of other lymphatic markers. Interestingly, a distinct local cytokine milieu, with decreased VEGF-A (P=0.04) and VEGF-D (P=0.02) expression was identified. No apparent lymphatic anomaly underlies lipedema, providing evidence for the different disease nature in comparison to lymphedema. The changes in the lymphatic-related cytokine milieu might be related to a modified vascular permeability developed secondarily to lipedema progression

A Distinct Cytokine Profile and Stromal Vascular Fraction Metabolic Status without Significant Changes in the Lipid Composition Characterizes Lipedema

Lipedema is an adipose tissue disorder characterized by the disproportionate increase of subcutaneous fat tissue in the lower and/or upper extremities. The underlying pathomechanism remains unclear and no molecular biomarkers to distinguish the disease exist, leading to a large number of undiagnosed and misdiagnosed patients. To unravel the distinct molecular characteristic of lipedema we performed lipidomic analysis of the adipose tissue and serum of lipedema versus anatomically- and body mass index (BMI)-matched control patients. Both tissue groups showed no significant changes regarding lipid composition. As hyperplastic adipose tissue represents low-grade inflammation, the potential systemic effects on circulating cytokines were evaluated in lipedema and control patients using the Multiplex immunoassay system. Interestingly, increased systemic levels of interleukin 11 (p = 0.03), interleukin 28A (p = 0.04) and interleukin 29 (p = 0.04) were observed. As cytokines can influence metabolic activity, the metabolic phenotype of the stromal vascular fraction was examined, revealing significantly increased mitochondrial respiration in lipedema. In conclusion, despite sharing a comparable lipid profile with healthy adipose tissue, lipedema is characterized by a distinct systemic cytokine profile and metabolic activity of the stromal vascular fraction

Jejunum free flap in hypopharynx reconstruction: Case series

BACKGROUND: Surgical treatment of hypopharyngeal cancers with extension to the retrocricoid region generally requires a circumferential pharyngolaryngectomy followed by a reconstruction of the removed segment of the upper digestive tract. Historically, many techniques have been used in order to achieve a safe and functional reconstruction. Jejunum interposition is generally considered the best reconstructive technique. METHODS: This study examines the details of the surgical technique, the complications, the oncological and the functional results in a series of 29 consecutive patients submitted to circumferential pharyngoesophageal resection for advanced hypopharyngeal cancer followed by reconstruction with a free flap of jejunum. RESULTS: Three of the transplants failed because of venous thrombosis. The overall success rate was 90%. There were no general complications. A good swallowing has been preserved in all our patients. All our patients where a phonatory prosthesis was positioned (20/29) were able to achieve speech following speech therapy and all were satisfied with their own capacity to communicate. CONCLUSIONS: The prognosis of hypopharyngeal tumours (18–40% at 5 years) remains poor, but jejunum autografts are being shown to be an excellent choice for the reconstruction of the cervical hypopharyngo-oesophagus offering the patient fast rehabilitation and a reasonable quality of survival. Our experience confirm that this kind of reconstruction is safe with a good results in improving oncologic controls and restoring a good quality of life