Loss of phospholipase PLAAT3 causes a mixed lipodystrophic and neurological syndrome due to impaired PPARγ signaling.

Phospholipase A/acyltransferase 3 (PLAAT3) is a phospholipid-modifying enzyme predominantly expressed in neural and white adipose tissue (WAT). It is a potential drug target for metabolic syndrome, as Plaat3 deficiency in mice protects against diet-induced obesity. We identified seven patients from four unrelated consanguineous families, with homozygous loss-of-function variants in PLAAT3, who presented with a lipodystrophy syndrome with loss of fat varying from partial to generalized and associated with metabolic complications, as well as variable neurological features including demyelinating neuropathy and intellectual disability. Multi-omics analysis of mouse Plaat3-/- and patient-derived WAT showed enrichment of arachidonic acid-containing membrane phospholipids and a strong decrease in the signaling of peroxisome proliferator-activated receptor gamma (PPARγ), the master regulator of adipocyte differentiation. Accordingly, CRISPR-Cas9-mediated PLAAT3 inactivation in human adipose stem cells induced insulin resistance, altered adipocyte differentiation with decreased lipid droplet formation and reduced the expression of adipogenic and mature adipocyte markers, including PPARγ. These findings establish PLAAT3 deficiency as a hereditary lipodystrophy syndrome with neurological manifestations, caused by a PPARγ-dependent defect in WAT differentiation and function.

Vascularized adipose tissue engineering: moving towards soft tissue reconstruction.

Soft tissue defects are a common clinical challenge mostly caused by trauma, congenital anomalies and oncological surgery. Current soft tissue reconstruction options include synthetic materials (fillers and implants) and autologous adipose tissue transplantation through flap surgery and/or lipotransfer. Both reconstructive options hold important disadvantages to which vascularized adipose tissue engineering (VATE) strategies could offer solutions. In this review, we first summarized pivotal characteristics of functional adipose tissue such as the structure, function, cell types, development and extracellular matrix (ECM). Next, we discussed relevant cell sources and how they are applied in different state-of-the-art VATE techniques. Herein, biomaterial scaffolds and hydrogels, ECMs, spheroids, organoids, cell sheets, three dimensional printing and microfluidics are overviewed. Also, we included extracellular vesicles and emphasized their potential role in VATE. Lastly, current challenges and future perspectives in VATE are pointed out to help to pave the road towards clinical applications.

The process to obtain reimbursement for enzymatic debridement in clinically deep burns.

INTRODUCTION: Burn eschar removal by enzymatic debridement with NexoBrid® (EDNX) results in a maximum preservation of all viable tissue, which is the main advantage over traditional tangential excision. The authors participated in a marketing authorization holder process to obtain reimbursement from the national health authorities in Belgium. MATERIAL AND METHODS: The reimbursement process consisted of three phases, as specified by the reimbursement regulations required by the Belgian National Institute for Health and Disability Insurance (NIHDI). RESULTS: Forty-one patients with clinically deep 2nd and 3rd degree burns, treated with EDNX in two Belgian burn centers, were included in the registry for the first phase of the reimbursement process. The total success rate of the EDNX treatment was 95.1% (39/41). Over half of the burn wounds treated with NexoBrid® (55.2%) did not require any additional surgical debridement or skin grafting. To obtain definitive reimbursement, an extra 16 patients were included. In this population, 51.4% did not require any surgical intervention. The total success rate of the EDNX debridement in this group was 100%. Based on an estimated market share of 12% and around 75 patients in the third year after final reimbursement, a market access consultant calculated that NexoBrid® will realize yearly savings for the Belgian Healthcare budget of at least €30.000. CONCLUSION: Based on the results of this Belgian registry study in combination with the yearly healthcare budget savings, the NIHDI granted a final reimbursement for EDNX treatment in adults, endorsed by the Minister of Health on November 5th, 2019.

Helpful hints in deciding what and when to operate after enzymatic debridement.

INTRODUCTION: In recent years, it has become clear that the burn eschar in deep burns can be selectively removed using the enzymatically debriding agent NexoBrid® (EDNX). In deep partial-thickness burns, such selective debridement preserves all non-injured dermis, which is sometimes sufficient for spontaneous re-epithelization. Nevertheless, it can be extremely challenging to determine exactly what and when to operate after an EDNX procedure. In this manuscript, we sought to investigate the clinical aspect of the enzymatically debrided wound bed of laser Doppler imaging (LDI)-confirmed deep dermal and full-thickness burns after NexoBrid® application. This to evaluate the residual wound healing capacity and implement specific indications for surgical therapy after enzymatic debridement. MATERIAL AND METHODS: Mainly LDI-blue areas, determined between 48 h and 5d after burn and afterwards treated with EDNX were selected. Six practical and three expert EDNX users evaluated the high-quality digital images of the wound beds immediately post NexoBrid® removal and after a 2 h wet-to-dry (WTD) dressing period. RESULTS: One hundred and two mainly LDI-blue areas in 32 patients were analyzed. Regarding the early decision-making, there were no significant differences in the wound bed evaluations, wound healing assessment and treatment decision of all 9 EDNX users post EDNX removal versus post WTD. Moreover, there was a good to excellent consensus between the practical and expert EDNX users in the individual wound bed evaluations. Even in the evaluation of a newly developed wound bed color code, with 7 different colors/patterns to choose from, the consensus was 80%. There was also an 84% consensus on the decision whether or not to operate. All mainly LDI-blue areas with incomplete enzymatic debridement, determined during clinical investigation by expert EDNX users, required surgery. Additionally, the expert investigators demonstrated that the following wound bed characteristics were independent predictors of the need for surgical treatment: visible fat lobules (p = 0.028), translucent fat lobules (p < 0.001), dermal step-off in the wound bed (p < 0.001), visible blood vessels (p < 0.001) and coagulated blood vessels (p = 0.023). Also, higher color code ranges on our own developed wound bed classification were significantly related to a surgical intervention (p = 0.006). When including the LDI flux values, the perfusion units were significantly different (p < 0.001) between the exclusively LDI-blue areas treated conservatively (mean 145.7) and the areas ultimately treated with autografts (mean 119.5). CONCLUSION: To the best of our knowledge, this study is the first to address the clinical wound bed evaluation of LDI-confirmed deep burns after NexoBrid® application. Based on our results, it is recommended to evaluate the wound bed twice: immediately after removing NexoBrid® to assess the viability of the wound bed and after the WTD period to reach a more complete decision. During these evaluations, wound bed characteristics such as incomplete debridement, visible and/or translucent fat lobules, visible and/or coagulated blood vessels and a dermal step-off in the wound bed combined with a higher range (4-5) in the newly developed wound bed color code should lead to an early and reliable decision for skin grafting. For burn centers using LDI, mean flux values below 119.5PU - in addition to the above-mentioned wound bed evaluation - are a clear indicator for surgical therapy.

Fat Embolism After Autologous Facial Fat Grafting.

BACKGROUND: Autologous facial fat grafting has gained popularity in recent years and is considered to be safe. This paper presents the case of a patient who died due to massive cerebral microfat embolism after facial fat grafting. OBJECTIVES: The aim of this study was to raise awareness and provide more evidence on the prevention and treatment of this potentially lethal complication of facial fat grafting. METHODS: A detailed report was made of the case. Two online databases were searched for similar cases of facial fat embolism resulting in neurologic and/or visual symptoms. Thereafter a literature search was conducted to verify the etiology, current treatment options, and preventive measures. RESULTS: Forty-nine cases with similar events were found in the literature. The most common injected area was the glabella (36.1%), and an average of 16.7 mL fat was injected. The main complications were visual impairment, with 88.5% of cases resulting in permanent monocular blindness, and neurologic symptoms, some of which never fully recovered. Including the present patient, 7 cases were fatal. Fat embolism can occur in the veins and arteries of the face. Two possible pathways for fat embolism exist: the macroscopic, mechanical pathway with immediate signs, and the microscopic, biochemical pathway with delayed symptoms. Mechanical embolectomy and corticosteroids are suggested treatment options but evidence for their efficacy is lacking. Several different preventive measures are described. CONCLUSIONS: Although facial fat grafting is considered a safe procedure, one should be aware of the risk of fat embolism. Underreporting of this adverse event is likely. With no effective treatment and often detrimental outcomes, preventive measures are of utmost importance to improve patient safety.

A Novel Technique for Preventive Lymphovenous Anastomosis: Anastomosing a Ligated Lymphatic Vessel.

BACKGROUND: Lymphedema is defined as a chronic condition, caused by lymphostasis. A major part in the Western world consists of iatrogenic lymphedema caused by surgery to the lymph nodes of the axilla or groin. Prophylactic lymphovenous anastomosis (LVA) could be beneficial in the prevention of lymphedema of the extremities. These procedures require experienced supramicrosurgeons and can be time consuming, which might be the reasons why prophylactic LVA has not yet been widely implemented in the treatment of cancer. Due to the small diameter of lymphatic vessels, it remains challenging to identify the lumen, and therefore, anastomoses are prone to back wall stitching. Different inventive procedures have been described making use of stents or monofilament sutures. METHODS: In this article, we describe a newly developed and straightforward technique for LVA in 4 patients who underwent an axilla dissection and 1 patient who underwent a dissection of the groin lymph nodes. This latter approach makes use of clipping of the lymphatic vessel during lymph node dissection, and remains ligated during anastomosis. The candidate vein was the V. thoracodorsalis for the axilla and the V. circumflexa superficialis for the groin. We describe the feasibility, average duration, and complications. RESULTS: Performing an end-to-side anastomosis on a clipped lymphatic vessel minimizes the problem of back wall stitching as well as the trouble of finding the lumen due to collapsing of the vessel. The turgor of the lymphatic vessel is maintained and makes anastomosing easy. Average time for LVA was 33.4 minutes and 1 minor complication was seen. CONCLUSION: We believe that this approach might be of value in popularizing LVA in the treatment or prevention of different conditions such as breast cancer-related lymphedema.

Nipple reconstruction in autologous breast reconstruction after areola-sparing mastectomy.

INTRODUCTION: The number of patients requesting prophylactic mastectomy with immediate reconstruction is rising. The oncological safety of techniques preserving the nipple and/or areola complex is still controversial. Nevertheless, nipple-sparing mastectomy (NSM) and areola-sparing mastectomy (ASM) are becoming increasingly popular. After ASM, traditional nipple reconstruction techniques can be a disappointment and can lead to a deep groove around the new nipple. We describe a technique to overcome these issues and analyzed how three types of mastectomy (skin-sparing mastectomy or SSM, ASM, and NSM) compare to one another by looking into the number of wound infections, extra procedures for the loss of projection, nipple necrosis, and BREAST-Q scores. METHODS: We retrospectively analyzed 467 breast reconstructions performed in 351 patients between 2011 and 2017 at the University Hospital of Gent. Patients were asked to fill out the BREAST-Q questionnaire and patient-reported outcomes were analyzed and correlated to demographic information. RESULTS: Patients undergoing a nipple reconstruction after ASM are experiencing similar rates of wound problems, extra surgical procedures for the loss of projection and necrosis, compared to women with a history of SSM. When considering the "satisfaction with breast" and "satisfaction with outcome" modules of the BREAST-Q, we noted that nipple-sparing mastectomy (NSM) patients report lower scores than SSM and ASM patients and ASM patients seem to report a higher "satisfaction with nipple," than the other two treatment groups. CONCLUSION: An ASM is a valuable alternative to a nipple-sparing mastectomy and leads to a good esthetic result and patient satisfaction.

Lumbar Flap versus the Gold Standard: Comparison to the DIEP Flap.

BACKGROUND: The lumbar artery perforator flap is an excellent free flap for breast reconstruction whenever the deep inferior epigastric perforator (DIEP) flap is not an option. The main indication is a lack of abdominal bulk, often seen in young BRCA-positive women seeking prophylactic amputation and immediate reconstruction. METHODS: Between October of 2010 and July of 2016, a total of 661 free flap breast reconstructions were performed. The authors retrospectively analyzed patient demographics, perioperative parameters, and secondary corrections. RESULTS: Seventy-six lumbar artery perforator flaps were retained and compared with a cohort of 560 DIEP flaps. The average body mass index for lumbar patients was 23.8 kg/m, with a mean age at operation of 46.3 years. Average body mass index for DIEP patients was 25.2 kg/m, with a mean age at operation of 48.8 years old. Lumbar artery perforator flap weight was 504 g (range, 77 to 1216 g) on average versus 530 g (range, 108 to 1968 g) for the DIEP flaps. The amount of corrective procedures performed was very similar in both cohorts: 13 percent of the lumbar artery perforator and 12 percent of the DIEP patients underwent no procedures, 62 percent in both groups underwent one procedure, and 25 percent versus 27 percent underwent two or more procedures. Lipofilling was performed in 48 percent of lumbar artery perforator flaps compared with 57 percent of the DIEP flaps (p = 0.14). Mean volume injected was 98.0 cc and 125.1 cc for lumbar artery perforator and DIEP flaps, respectively (p = 0.071). CONCLUSIONS: The lumbar flap is a good alternative whenever a DIEP flap is not possible. Bilateral autologous reconstruction is possible even in very thin patients, and secondary corrections are comparable to those for the DIEP.

Author Correction: 20 Years of DIEAP Flap Breast Reconstruction: A Big Data Analysis.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

20 Years of DIEAP Flap Breast Reconstruction: A Big Data Analysis.

With every hospital admission, a vast amount of data is collected from every patient. Big data can help in data mining and processing of this volume of data. The goal of this study is to investigate the potential of big data analyses by analyzing clinically relevant data from the immediate postoperative phase using big data mining techniques. A second aim is to understand the importance of different postoperative parameters. We analyzed all data generated during the admission of 739 women undergoing a free DIEAP flap breast reconstruction. The patients' complete midcare nursing report, laboratory data, operative reports and drug schedule were examined (7,405,359 data points). The duration of anesthesia does not predict the need for revision. Low Red Blood cell Counts (3.53 × 106/µL versus 3.79 × 106/µL, p < 0.001) and a low MAP (MAP = 73.37 versus 76.62; p < 0.001) postoperatively are correlated with significantly more revisions. Different drugs (asthma/COPD medication, Butyrophenones) can also play a significant role in the success of the free flap. In a world that is becoming more data driven, there is a clear need for electronic medical records which are easy to use for the practitioner, nursing staff, and the researcher. Very large datasets can be used, and big data analysis allows a relatively easy and fast interpretation all this information.

Breast levonorgestrel concentrations in women using a levonorgestrel-releasing intrauterine system.

OBJECTIVE: To measure breast tissue and serum LNG concentrations in women using a LNG-IUS. STUDY DESIGN: This pilot study was performed in 25 healthy women undergoing breast surgery at the Ghent University hospital. LNG concentrations were measured in serum and microdissected breast tissue samples using a validated ultra-performance liquid chromatography/tandem mass spectrometry assay. RESULT(S): The mean LNG concentration in the 18 LNG-IUS users was 0.18±0.16 ng/mL in serum and 0.26±0.28 ng/g in breast tissue. For four women without any form of hormonal contraceptive (the negative controls), the mean concentrations were below the limit of quantification, i.e., 0.15 ng/mL and 0.20 ng/g, for serum and breast tissue, respectively. For the three positive controls the concentrations in the serum (20.5 and 3.4 ng/ml) and the breast (3.74 and 1.24 ng/g) were respectively for the 20 µg EE/100 µg users and 315 pg/ml in the serum and 1.17 ng/g in the breast for the minipill user. The intracellular free fraction of LNG may be as low as 0.008 ng/g. CONCLUSION(S): The concentration of LNG in breast epithelium cells in women using the LNG-IUS is very low. IMPLICATIONS: The relationship between the serum and breast tissue levels of LNG was studied in women using a LNG-IUS or oral LNG-containing contraception. Compared to oral contraception, the tissue levels of LNG in LNG-IUS users are much lower in the breast. It is not known what level of LNG exposure in the breast would stimulate RANKL and WNT4 expression; such information is needed.

Autologous plasma and its supporting role in fat graft survival: A relevant vector to counteract resorption in lipofilling.

Fat grafting has become a widespread technique for different reconstructive and esthetic purposes. However, the disadvantage of fat grafting is the unpredictable resorption rate that often necessitates repetitive procedures, which in turn may have an impact on the morbidity. During the immediate, post-graft, ischemic period, cells survive due to the process of plasmatic imbibition. This biological phenomenon precedes the ingrowth of neo-capillaries that eventually nourish the graft and help establish a long-term homeostatic equilibrium. Both partners, the graft and the recipient bed, contribute to the revascularization process. Hypothetically, enrichment of the recipient site with autologous plasma could have a beneficial role to enhance fat graft survival. We investigated whether plasma supported the viability of the lipoaspirate (LA) material. Plasma was isolated from blood samples collected from eight patients during the elective lipofilling procedures. An in vitro study assessed the viability of LA cells using plasma as a culture medium compared to the traditional culture media. In vitro analysis confirmed sustained viability of LA cells compared to the standard media and control media during 7 consecutive days. The behavior of the fat grafts in plasma showed similarities with those incubated in the traditional culture media. In future, these findings could be translated to a clinical setting. Plasma is the only autologous substrate available in large quantities in the human body. The addition of the supporting agents, such as plasma, could contribute to a better graft survival with more stable clinical outcomes in the long term. The rationale behind the technique is based on the phenomenon of plasmatic imbibition and the reasoning that the extracellular matrix plays a pivotal role in cellular survival.

The dog-ear flap as an alternative for breast reconstruction in patients who have already undergone a DIEAP flap.

Breast reconstruction in patients who have previously undergone deep inferior epigastric artery perforator flap (DIEAP) reconstruction or abdominoplasty is often challenging. Depending on patients' body habitus, several second-choice flaps have been described such as the transverse upper gracilis (TUG) flap, profundus femoris artery perforator (PFAP) flap, superior gluteal artery perforator (SGAP) flap, and lumbar artery perforator (LAP) flap. Patients who have undergone a DIEAP flap reconstruction or abdominoplasty occasionally present with dog ears on both sides of the abdominal scar. The adipose tissue and skin of these dog ears are supplied by perforators of the deep circumflex iliac artery (DCIA). The DCIA flap was first described in 1979 by Taylor. We introduce this abdominal "dog-ear" flap for autologous breast reconstruction.

Intrinsic dynamics of the fat graft: in vitro interactions between the main cell actors.

BACKGROUND: Successful soft-tissue reconstruction requires autologous tissue transfer in respect to the increasingly important "replace like-with-like" principle. Autologous lipoaspirate material for fat grafting can easily be obtained in large amounts without substantial donor-site morbidity. The exact nature and fate of the different cells in the transplanted fat graft and their contribution to tissue reconstruction, however, remain largely unknown. METHODS: Adipose tissue was harvested from healthy female patients. CD34+ adipose-derived stem cells were isolated through magnetic-activated cell sorting and brought into co-culture with mature adipocytes in various culture medium conditions. Proliferation and differentiation of the adipose-derived stem cells were examined through histology, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, and polymerase chain reaction assays. RESULTS: This study demonstrates that adipose-derived stem cells from fresh adipose tissue can be isolated within a few hours via magnetic-activated cell sorting with selection for CD34+ cells. All unpassaged adipose-derived stem cells in fresh adipose tissue are CD34+. Subsets include CD34+ CD31+ and CD34+ CD271+. No CD34+ CD45+ cells were present. Histological staining, polymerase chain reaction, and MTT assays confirm that purified mature adipose cells incite adipose-derived stem cells proliferation and adipose differentiation in vitro. CONCLUSIONS: This in vitro study demonstrates important interactions between the main actors in the adipose graft, the adipose-derived stem cells and the mature adipocytes. Although the eventual fate of these cells in a clinically implemented fat graft is still largely unknown, the results of this study support the theory that lipofilling can be conceived as an in vivo tissue engineering approach in which the mature adipocytes within fat grafts support proliferation and differentiation in the co-grafted stromal cell population.