The Triple Adipose-Derived Stem Cell Exosome Technology as a Potential Tool for Treating Triple-Negative Breast Cancer.

BACKGROUND: Extracellular vesicles are pivotal mediators in intercellular communication, facilitating the exchange of biological information among healthy, pathological and tumor cells. Between the diverse subtypes of extracellular vesicles, exosomes have unique properties and clinical and therapeutical applications. Breast cancer ranks as one of the most prevalent malignancies across the globe. Both the tumor core and its surrounding microenvironment engage in a complex, orchestrated interaction that facilitates cancer's growth and spread. METHODS: The most significant PubMed literature about extracellular vesicles and Adipose-Derived Stem Cell Exosomes and breast cancer was selected in order to report their biological properties and potential applications, in particular in treating triple-negative breast cancer. RESULTS: Adipose-Derived Stem Cell Exosomes represent a potential tool in targeting triple-negative breast cancer cells at three main levels: the tumor core, the tumor microenvironment and surrounding tissues, including metastases. CONCLUSIONS: The possibility of impacting triple-negative breast cancer cells with engineered Adipose-Derived Stem Cell Exosomes is real. The opportunity to translate our current in vitro analyses into a future in vivo scenario is even more challenging.

Exosomes from Adipose-Tissue-Derived Stem Cells Induce Proapoptotic Gene Expression in Breast Tumor Cell Line.

Lipofilling is an option for breast reconstruction after tumor resection to avoid the complications of an implant-based reconstruction. Although some concerns exist regarding the oncological safety of tissue rich in mesenchymal stem cells with their proangiogenic and proliferation-supportive properties, there are also reports that adipose-tissue-derived stem cells can exhibit antitumoral properties. We isolated primary adipose-tissue-derived stem cells. Both conditioned medium and exosomes were harvested from the cell culture and used to treat the breast cancer cell line MCF-7. Cell viability, cytotoxicity, and gene expression of MCF-7 cells in response to the indirect co-culture were evaluated. MCF-7 cells incubated with exosomes from adipose-tissue-derived stem cells show reduced cell viability in comparison to MCF-7 cells incubated with adipose-tissue-derived stem-cell-conditioned medium. Expression of proapoptotic genes was upregulated, and expression of antiapoptotic genes was downregulated. The debate about the oncological safety of autologous fat grafting after tumor resection continues. Here, we show that exosomes from adipose-tissue-derived stem cells exhibit some antitumoral properties on breast cancer cell line MCF-7.

Evidence-Based Approaches to Anticoagulation in Reconstructive Microsurgery-A Systematic Literature Review.

This systematic review addresses the crucial role of anticoagulation in microsurgical procedures, focusing on free flap reconstruction and replantation surgeries. The objective was to balance the prevention of thrombotic complications commonly leading to flap failure, with the risk of increased bleeding complications associated with anticoagulant use. A meticulous PubMed literature search following Evidence-Based-Practice principles yielded 79 relevant articles, including both clinical and animal studies. The full-texts were carefully reviewed and evaluated by the modified Coleman methodology score. Clinical studies revealed diverse perioperative regimens, primarily based on aspirin, heparin, and dextran. Meta-analyses demonstrated similar flap loss rates with heparin or aspirin. High doses of dalteparin or heparin, however, correlated with higher flap loss rates than low dose administration. Use of dextran is not recommended due to severe systemic complications. In animal studies, systemic heparin administration showed predominantly favorable results, while topical application and intraluminal irrigation consistently exhibited significant benefits in flap survival. The insights from this conducted systematic review serve as a foundational pillar towards the establishment of evidence-based guidelines for anticoagulation in microsurgery. An average Coleman score of 55 (maximum 103), indicating low overall study quality, however, emphasizes the need for large multi-institutional, randomized-clinical trials as the next vital step.

The masseteric nerve for facial reanimation: Macroscopic and histomorphometric characteristics in 106 human cadavers and comparison of axonal ratio with recipient nerves.

Peripheral facial palsy causes severe impairments. Sufficient axonal load is critical for adequate functional outcomes in reanimation procedures. The aim of our study was to attain a better understanding of the anatomy of the masseteric nerve as a donor, in order to optimize neurotization procedures. Biopsies were obtained from 106 hemifaces of fresh frozen human cadavers. Histological cross-sections were fixed, stained with PPD, and digitized. Histomorphometry and a validated software-based axon quantification were conducted. Of the 154 evaluated branches, 74 specimens were of the main trunk (MT), 40 of the anterior branch (AB), and 38 of the descending branch (DB), while two halves of one cadaver featured an additional branch. The MT showed a diameter of 1.4 ± 0.41 mm (n = 74) with 2213 ± 957 axons (n = 55). The AB diameter was 0.9 ± 0.33 mm (n = 40) with 725 ± 714 axons (n = 30). The DB diameter was 1.15 ± 0.34 mm (n = 380) with 1562 ± 926 axons (n = 30). The DB demonstrated a high axonal capacity - valuable for nerve transfers or muscle transplants. Our findings should facilitate a balanced selection of axonal load, and are potentially helpful in achieving more predictable results while preserving masseter muscle function.

Histomorphometry of the Sural Nerve for Use as a CFNG in Facial Reanimation Procedures.

Facial palsy (FP) is a debilitating nerve pathology. Cross Face Nerve Grafting (CFNG) describes a surgical technique that uses nerve grafts to reanimate the paralyzed face. The sural nerve has been shown to be a reliable nerve graft with little donor side morbidity. Therefore, we aimed to investigate the microanatomy of the sural nerve. Biopsies were obtained from 15 FP patients who underwent CFNG using sural nerve grafts. Histological cross-sections were fixated, stained with PPD, and digitized. Histomorphometry and a validated software-based axon quantification were conducted. The median age of the operated patients was 37 years (5-62 years). There was a significant difference in axonal capacity decrease towards the periphery when comparing proximal vs. distal biopsies (p = 0.047), while the side of nerve harvest showed no significant differences in nerve caliber (proximal p = 0.253, distal p = 0.506) and axonal capacity for proximal and distal biopsies (proximal p = 0.414, distal p = 0.922). Age did not correlate with axonal capacity (proximal: R = -0.201, p = 0.603; distal: R = 0.317, p = 0.292). These novel insights into the microanatomy of the sural nerve may help refine CFNG techniques and individualize FP patient treatment plans, ultimately improving overall patient outcomes.

Precise Monitoring of Returning Sensation in Digital-Nerve Lesions by Three-Dimensional Imaging: A Proof-of-Concept Study.

SUMMARY: Digital-nerve lesions result in a loss of tactile sensation reflected by an anesthetic area (AA) at the radial or ulnar aspect of the respective digit. Available tools to monitor the recovery of tactile sense have been criticized for their lack of validity. Precise quantification of AA dynamics by three-dimensional (3D) imaging could serve as an accurate surrogate to monitor recovery after digital-nerve repair. For validation, AAs were marked on digits of healthy volunteers to simulate the AA of an impaired cutaneous innervation. The 3D models were composed from raw images that had been acquired with a 3D camera to precisely quantify relative AA for each digit (3D models, n = 80). Operator properties varied with regard to individual experience in 3D imaging and image processing. In addition, the concept was applied in a clinical case study. Results showed that images taken by experienced photographers were rated as better quality ( P < 0.001) and needed less processing time ( P = 0.020). Quantification of the relative AA was not altered significantly, regardless of experience level of the photographer ( P = 0.425) or image assembler ( P = 0.749). The proposed concept allows precise and reliable surface quantification of digits and can be performed consistently without relevant distortion by lack of examiner experience. Routine 3D imaging of the AA has the great potential to provide visual evidence of various returning states of sensation and to convert sensory nerve recovery into a metric variable with high responsiveness to temporal progress.

Standard doses of Triamcinolone do not affect fibroblast cell migration of abdominoplasty patients in-vitro1.

BACKGROUND: Recent studies have demonstrated that local application of corticosteroids reduces wound exudation following abdominoplasty and other reconstructive surgical procedures. On the other hand, corticosteroids might provoke wound healing disturbances due to their immunosuppressive effects. OBJECTIVE: The main objective of this study was to gain further information about the impact of the corticosteroid triamcinolone on cell migration in abdominoplasty patients. METHODS: An in-vitro scratch assay wound healing model was applied to observe cell migration of fibroblasts cultured with nutrient medium containing human seroma aspirate±triamcinolone. RESULTS: There were no significant differences regarding cell migration when fibroblasts were incubated with triamcinolone + seroma containing culture medium compared to seroma containing culture medium without triamcinolone. CONCLUSIONS: The performed in-vitro study suggests that triamcinolone does not decelerate fibroblast cell migration which is considered as a surrogate of wound healing.

Peripheral Nerve Regeneration-Adipose-Tissue-Derived Stem Cells Differentiated by a Three-Step Protocol Promote Neurite Elongation via NGF Secretion.

The lack of supportive Schwann cells in segmental nerve lesions seems to be one cornerstone for the problem of insufficient nerve regeneration. Lately, adipose-tissue-derived stem cells (ASCs) differentiated towards SC (Schwann cell)-like cells seem to fulfill some of the needs for ameliorated nerve recovery. In this study, three differentiation protocols were investigated for their ability to differentiate ASCs from rats into specialized SC phenotypes. The differentiated ASCs (dASCs) were compared for their expressions of neurotrophins (NGF, GDNF, BDNF), myelin markers (MBP, P0), as well as glial-marker proteins (S100, GFAP) by RT-PCR, ELISA, and Western blot. Additionally, the influence of the medium conditioned by dASCs on a neuron-like cell line was evaluated. The dASCs were highly diverse in their expression profiles. One protocol yielded relatively high expression rates of neurotrophins, whereas another protocol induced myelin-marker expression. These results were reproducible when the ASCs were differentiated on surfaces potentially used for nerve guidance conduits. The NGF secretion affected the neurite outgrowth significantly. It remains uncertain what features of these SC-like cells contribute the most to adequate functional recovery during the different phases of nerve recovery. Nevertheless, therapeutic applications should consider these diverse phenotypes as a potential approach for stem-cell-based nerve-injury treatment.

Technical Aspects of High-Resolution Color-Coded Duplex Sonography for the Design of Perforator Flaps.

BACKGROUND: Technical aspects are of utmost significance for an efficient execution in designing perforator flaps with high-resolution color-coded Duplex sonography (CCDS). The following study evaluates decisive factors for a successful microvessel examination conducted by the microsurgeon. METHODS: Technical knowledge presented in this study was based on a series of more than 200 perforator flaps planned with CCDS. Flap reconstructions were performed at the University Hospital Regensburg, Germany, from July 2013 to January 2021. Standard high-resolution ultrasound (US) devices with linear multifrequency transducers of 4 to 18 MHz were used. Modes and device settings were evaluated regarding applicability by microsurgeons. Key steps for safe perforator identification and further optional steps for additional assessment should be discriminated. RESULTS: Different US modes including brightness mode (B-mode), color flow (CF), power Doppler (PD), pulse wave (PW), and blood flow (B-Flow) were used. Transducers from 15 MHz and up were favorable to detect microvessels. Knobology of a standard US device regarding buttons, switches, and specific onscreen options with relevance for perforator mapping was subcategorized in four different groups. For qualitative and quantitative evaluation of microvessels, different US modes were tested with respect to their usefulness.Vital elements of the CCDS exam are disaggregated into three key steps for safe perforator identification and three optional steps for further perforator characterization. A standardized protocol for the CCDS exams was applied. Downregulation of pulse-repetition frequency/scale to adapt device sensitivity to slow-flow velocities represented the most important criterion to visualize microvessels.Qualitative microvessel evaluation was performed in B-mode, CCDS, PD mode, and B-Flow mode. Quantitative assessment was executed using PW-mode and CCDS measuring the microvessels' diameter (mm) and flow characteristics. Quantitative information may be obtained using PW-mode and the distance-measuring tool in CF-mode. CONCLUSION: Technical aspects with respect to proper device trimming and application decisively impact CCDS-guided perforator vessel identification and evaluation.

Monitoring free flaps and replanted digits via perfusion index - A proof of concept study.

BACKGROUND: Early detection and treatment of vascular complications in replanted digits is essential for the survival. The perfusion index (PI) represents a marker of peripheral perfusion as it shows the ratio of pulsatile to non-pulsatile blood flow. OBJECTIVE: To evaluate the feasibility and applicability of the PI as a monitoring tool for free flaps and replanted digits by measuring the inter- and intraindividual changes in PI. METHODS: Five patients were postoperatively monitored according to intern standards by hourly clinical evaluation. Additionally, a pulse oximeter with SET-technology® (Masimo Radial 7, Masimo Corporation, Irvine, USA) was added with a LNCS® Red TFA-1 SpO2 sensor (Masimo Corporation, Irvine, USA) and respectively a LNCS® Neo-3 neonatal finger clip to evaluate the perfusion via PI and SpO2. RESULTS: All patients showed sufficient perfusion in clinical controls. There was no detectable vascular complication during follow-up. Mean perfusion index was 0.93 with a median of 0.44. The patients showed a mean SpO2 of 90.59%with a median of 89.21%. CONCLUSION: Our results show a great intra- and interindividual range of PI and SpO2. SpO2 provided an even greater range than PI. Trends in intraindividual PI changes may be a promising monitoring tool for free flaps and replanted digits.

"A-PePSI LIGhT" Assessment Score to Predict Pressure Sore Impaired Healing Late Recurrence, Immobility, Greater Surface, Inhibited Thrombocytes.

BACKGROUND: Complication rates of up to 46 percent are reported following pressure sore surgery. Pressure sore patients often exhibit ineffective postoperative wound healing despite tension-free flap coverage, necessitating surgical revision and prolonged hospitalization. Rather than pressure sore recurrence, such impaired healing reflects a failed progress through the physiologic stages of the normal wound-healing cascade. The principal objective of the study reported here was to elucidate potentially modifiable inherent variables that predict predisposition to impaired healing and to provide a tool for identifying cases at risk for complicated early postoperative recovery following pressure sore reconstruction. METHODS: A retrospective chart review of late-stage (stage 3 or higher) sacral and ischial pressure sore patients who underwent flap reconstruction from 2014 to 2019 was performed. A multivariable logistic regression model was used to identify key patient and operative factors predictive of impaired healing. Furthermore, the Assessment Score to Predict Pressure Sore Impaired Healing (A-PePSI) was established based on the identified risk factors. RESULTS: In a cohort of 121 patients, 36 percent exhibited impaired healing. Of these, 34 patients suffered from dehiscences, necessitating surgical revision. Statistically significant risk factors comprising late recurrence (OR, 3.8), immobility (OR, 12.4), greater surface (>5 cm diameter; OR, 7.3), and inhibited thrombocytes (aspirin monotherapy; OR, 5.7) were combined to formulate a prognostic scoring system (A-PePSI LIGhT). CONCLUSIONS: The A-PePSI LIGhT score serves as a prognostic instrument for assessing individual risk for impaired healing in pressure sore patients. Preoperative risk stratification supports rational decision-making regarding operative candidacy, allows evidence-based patient counseling, and supports the implementation of individualized treatment protocols. . CLINICAL QUESTION/LEVEL OF EVIDENCE: Risk, III.

Microanatomy of the Frontal Branch of the Facial Nerve: The Role of Nerve Caliber and Axonal Capacity.

BACKGROUND: A commonly seen issue in facial palsy patients is brow ptosis caused by paralysis of the frontalis muscle powered by the frontal branch of the facial nerve. Predominantly, static methods are used for correction. Functional restoration concepts include the transfer of the deep temporal branch of the trigeminal nerve and cross-facial nerve grafts. Both techniques can neurotize the original mimic muscles in early cases or power muscle transplants in late cases. Because axonal capacity is particularly important in cross-facial nerve graft procedures, the authors investigated the microanatomical features of the frontal branch to provide the basis for its potential use and to ease intraoperative donor nerve selection. METHODS: Nerve biopsy specimens from 106 fresh-frozen cadaver facial halves were obtained. Histologic processing and digitalization were followed by nerve morphometric analysis and semiautomated axon quantification. RESULTS: The frontal branch showed a median of three fascicles (n = 100; range, one to nine fascicles). A mean axonal capacity of 1191 ± 668 axons (range, 186 to 3539 axons; n = 88) and an average cross-sectional diameter of 1.01 ± 0.26 mm (range, 0.43 to 1.74 mm; n = 67) were noted. In the linear regression model, diameter and axonal capacity demonstrated a positive relation (n = 57; r2 = 0.32; p < 0.001). Based on that equation, a nerve measuring 1 mm is expected to carry 1339 axons. CONCLUSION: The authors' analysis on the microanatomy of the frontal branch could promote clinical use of cross-facial nerve graft procedures in frontalis muscle neurotization and free muscle transplantations.

Do-It-Yourself Preoperative High-Resolution Ultrasound-Guided Flap Design of the Superficial Circumflex Iliac Artery Perforator Flap (SCIP).

The superficial circumflex iliac artery perforator (SCIP) flap is a well-documented, thin, free tissue flap with a minimal donor site morbidity, and has the potential to become the new method for resurfacing moderate-size skin defects. The aim of this study is to describe an easy, reliable, systematic, and standardized approach for preoperative SCIP flap design and perforator characterization, using color-coded duplex sonography (CCDS). A list of customized settings and a straightforward algorithm are presented, which are easily applied by an operator with minimal experience. Specific settings for SCIP flap perforator evaluation were investigated and tested on 12 patients. Deep and superficial superficial circumflex iliac artery (SCIA) branches, along with their corresponding perforators and cutaneous veins, were marked individually with a permanent marker and the anatomy was verified intraoperatively. From this, a simplified procedure for preoperative flap design of the SCIP flap was developed. Branches could be localized and evaluated in all patients. A preoperative structured procedure for ultrasonically guided flap design of the SCIP flap is described. A 100% correlation between the number and emergence points of the branches detected by preoperative CCDS mapping and the intraoperative anatomy was found.

Local Triamcinolone Treatment Affects Inflammatory Response in Seroma Exudate of Abdominoplasty Patients: A Randomized Controlled Trial.

BACKGROUND: As the leading complication of abdominoplasty, seroma formation might represent an inflammatory process in response to surgical trauma. This prospective randomized trial investigated whether local administration of the antiinflammatory agent triamcinolone could prevent seroma accumulation. METHODS: Weekly and cumulative seroma volumes were compared between the study groups A, B, and C over a 4-week follow-up (group A, with drain, without triamcinolone; group B, without drain, without triamcinolone; group C, without drain, with triamcinolone). Aspirated seroma samples were analyzed by enzyme-linked immunosorbent assay for selective inflammatory mediators. RESULTS: Triamcinolone significantly reduced cumulative seroma volume (n = 60; mA 845 ± SDA 578 ml, mC 236 ± SDC 381 ml, p = 0.001). The most accentuated suppressive effect of triamcinolone was observed shortly after the treatment (week 1) (mA1 616 ± SDA1 457 ml, mB1 153 ± SDB1 161 ml, mC1 22 ± SDC1 44 ml, pA1/C1 < 0.001, pB1/C1 = 0.014). Local triamcinolone administration resulted in a differential concentration of interleukin-6 (IL-6) and matrix metalloproteinase-9 (MMP-9 (week 1) in seroma exudate as measured by enzyme-linked immunosorbent assay (mIL-6A1 1239 ± SDA1 59 pg/ml, mIL-6C1 848 ± SDC1 80 pg/ml, p < 0.001; mMMP-9A1 2343 ± SDA1 484 pg/ml, mMMP-9C1 376 ± SDC1 120 pg/ml, p = 0.001). CONCLUSIONS: Local administration of 80 mg of triamcinolone reduced postabdominoplasty seroma accumulation significantly. Under triamcinolone treatment, suppressed levels of IL-6 and MMP-9 in seroma fluid were observed. Notably, inflammatory marker suppression correlated clinically with a decrease in seroma accumulation. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, II.

High-Resolution Ultrasound-Guided Perforator Mapping and Characterization by the Microsurgeon in Lower Limb Reconstruction.

BACKGROUND: Preoperative ultrasound (US)-guided perforator mapping has immensely simplified perforator flap planning. It may be executed by the microsurgeon. Device settings and selection of ultrasound modes are of utmost significance for detection of low-flow microvessels. The following study evaluates different US modes. METHODS: A prospective complete data acquisition was performed from July 2018 to June 2019 in a subset of patients who underwent US-guided flap planning. Multifrequency linear transducers were used applying five US modes. Brightness (B)-mode, color flow (CF), power Doppler (PD), pulse wave (PW), and B-flow modes were evaluated regarding applicability by microsurgeons. Peak systolic velocity (PSV), end diastolic velocity (EDV), and resistance index (RI) were chosen to evaluate flow characteristics. US results were correlated to intraoperative findings. RESULTS: A total number of eight patients (six males and two females) undergoing anterolateral thigh (ALT) or superficial circumflex iliac artery perforator (SCIP) flap surgery received an extensive standardized US-guided perforator characterization. Qualitative evaluation was performed in B-mode, color-coded duplex sonography (CCDS), PD, and B-flow mode. Quantitative assessment was executed using PW-mode and CCDS measuring the microvessels' diameter (mm) and flow characteristics (PSV, EDV, and RI). CCDS provided a mean diameter of 1.93 mm (range: 1.2-2.8 ± 0.51), a mean systolic peak of 16.9 cm/s (range: 9.9-33.4 ± 7.79), and mean RI of 0.71 (range: 0.55-0.87 ± 0.09) for lower limb perforators. All perforators located with US were verified by intraoperative findings. An optimized, time-effective US mapping algorithm was derived. Qualitative parameters may be evaluated with B-mode, CF, or B-flow. Smallest microvessels may be assessed in PD-mode. Lowering pulse-repetition frequency (PRF)/scale is mandatory to image low-flow microvessels as perforators. Quantitative information may be obtained using PW-mode and the distance-measuring tool in CF-mode. Image and video materials are provided. CONCLUSION: CCDS proved to be a powerful tool for preoperative perforator characterization when using a structured approach and mapping algorithm. Different techniques may be applied for specific visualizations and performed by the microsurgeon.

[Abdominoplasty without closed-suction drains: a randomised controlled trial]. / Verzicht auf Drainagen bei der Abdominoplastik ­ eine randomisiert-kontrollierte Studie.

INTRODUCTION: There is insufficient scientific evidence from randomised controlled trials to support the routine use of closed-suction drains in body contouring procedures. The aim of this study was to evaluate cumulative seroma volume, length of hospital stay and complication rates in abdominoplasty patients without drains in direct comparison with a cohort receiving drains. MATERIAL AND METHODS: Abdominoplasty patients were prospectively randomised in two study groups with (MD) and without (OD) placement of closed-suction drains. Patients with a BMI ≤ 30 kg/m2 undergoing horizontal or combined horizontal/vertical incision abdominoplasty were included. Scarpa's fascia was preserved during dissection. Exclusion criteria comprised simultaneous liposuction, coagulation disorders and ASA score ≥ 3. Cumulative seroma volume over a four-week follow-up period was assessed as the primary outcome measure. Secondary outcome measures were complications requiring surgical revision and length of hospital stay. RESULTS: This trial did not identify a statistically significant difference in cumulative seroma volume between the MD (30/53) and OD (23/53) cohorts in 53 patients (MMD 493 ± SD 407 ml; MOD 459 ± SD 624 ml; p = 0.812). However, a significantly shorter average length of hospital stay was observed in the OD population (MMD 5.1 ± SD 1.4 d; MOD 4.2 ± SD 1.5 d; p = 0.023). Complication rates were equal in both study groups (nMD = 1; nOD = 1). CONCLUSION: The results of this trial do not justify routine placement of closed-suction drains in abdominoplasty procedures (horizontal or combined horizontal/vertical incision) in the pre-obese patient cohort (BMI ≤ 30 kg/m2). Drain placement should be evaluated on an individual patient-specific basis.

Shear Force Processing of Lipoaspirates for Stem Cell Enrichment Does Not Affect Secretome of Human Cells Detected by Mass Spectrometry In Vitro.

BACKGROUND: Lipofilling is one of the most often performed surgical procedures in plastic and reconstructive surgery. Lipoaspirates provide a ready source of stem cells and secreted factors that contribute to neoangiogenesis and fat graft survival. However, the regulations about the enrichment of these beneficial cells and factors are ambiguous. In this study, the authors tested whether a combination of centrifugation and homogenization allowed the enrichment of viable stem cells in lipoaspirates through the selective removal of tumescent solution, blood, and released lipids without significantly affecting the cell secretome. METHODS: Human lipoaspirate was harvested from six different patients using water jet-assisted liposuction. Lipoaspirate was homogenized by first centrifugation (3584 rpm for 2 minutes), shear strain (10 times intersyringe processing), and second centrifugation (3584 rpm for 2 minutes). Stem cell enrichment was shown by cell counting after stem cell isolation. Lipoaspirate from different processing steps (unprocessed, after first centrifugation, after homogenization, after second centrifugation) was incubated in serum-free cell culture medium for mass spectrometric analysis of secreted proteins. RESULTS: Lipoaspirate homogenization leads to a significant 2.6 ± 1.75-fold enrichment attributable to volume reduction without reducing the viability of the stem cells. Protein composition of the secretome did not change significantly after tissue homogenization. Considering the enrichment effects, there were no significant differences in the protein concentration of the 83 proteins found in all processing steps. CONCLUSIONS: Stem cells can be enriched mechanically without significantly affecting the composition of secreted proteins. Shear-assisted enrichment of lipoaspirate constitutes no substantial manipulation of the cells' secretome.

Clinical Impact of DIEP Flap Perforator Characteristics - A Prospective Indocyanine Green Fluorescence Imaging Study.

BACKGROUND: The question to what extent perfusion in deep inferior epigastric perforator (DIEP) flaps depends on specific perforator characteristics has been raised. Anatomical studies and previous clinical trials focussing on DIEP flap perfusion resulted in discrepancies. This prospective study investigates how perforator row, number and diameter affect DIEP flap microperfusion via Indocyanine Green (ICG) fluorescence angiography. METHODS: The fractional weight of insufficiently perfused flap tissue in Zone 4 related to the total DIEP flap weight was measured based on ICG fluorescence angiography and defined as Zone 4 %. As a surrogate for overall DIEP flap perfusion, Zone 4 % was assessed according to the row, number and diameter of perforators included in the flap. RESULTS: In 42 unilateral DIEP flap breast reconstructions, neither medial (33.6 ± 14.2 %)/lateral perforator row (29.9 ± 7.5 %, p = 0.683) nor the parameter perforator number (single perforator 31.5 ± 14.4 %, two perforators 30.2 ± 10.2 %, p = 0.727) had a statistically significant effect on flap tissue availability as measured via Zone 4 %. A negative correlative trend between perforator diameter and Zone 4 % (r = -0.096, p = 0.588) was observed. CONCLUSION: Zone 4 % provides a novel method for an objective assessment of DIEP flap perfusion. Medial/lateral row selection and other perforator properties (number, diameter) within the standard ranges, did not affect Zone 4 % as indicated by ICG fluorescence angiography.

Step-by-step guide to ultrasound-based design of alt flaps by the microsurgeon - Basic and advanced applications and device settings.

BACKGROUND: The anterolateral thigh (ALT) perforator flap is a popular reconstructive tissue transfer. Consistent with the "hot/cold zone" concept for rapid dissection and thin flap harvest, reliable preoperative perforator mapping is mandatory. Color-coded duplex sonography (CCDS) has been shown to have the highest pooled sensitivity and positive predictive value to identify ALT perforating vessels. By reviewing this guide, the reader should learn: 1. Probe selection and basic/advanced device settings 2. Interpreting tissue morphology 3. Structured mapping approach 4. Pedicle position planning 5. Safe flap design 6. Assess subcutaneous course and flap's thickness for subfascial/epifascial/suprafascial harvest 7. Implement perforators identified into a tailor-made flap design including chimeric flaps. METHODS: Experiences with ultrasound-guided flap design gained from 125 ALT perforator flap free tissue transfers performed in two reconstructive centers was the basis of our guide. Our structured method comprises standardized markings, patient positioning, and simple ergonomics. Basic and advanced CCDS settings, selection, and conventional probe guidance are outlined for the microsurgeon. RESULTS: Linear multifrequency probes (6-15 MHz) were used. Best preset programs were breast, thyroid, and vascular. Favorable device properties were depth focused to 2-5 cm, pulse repetition frequency (PRF/Scale) set low to 0.5-1.5 kHz/3-10 cm/s, color gain high, and wall filter (WF) low/off (< 50 Hz). Additional parameters were discussed. A 100% concordance rate was seen comparing preoperative perforator visualization with CCDS and intraoperative findings. Detailed picture and video material were demonstrated. CONCLUSION: CCDS is a powerful tool for preoperative perforator mapping in perforator flaps such as the ALT.