Plasma Cytokine and Growth Factor Profiling during Free Flap Transplantation

Ischemia and reperfusion (I/R) is an unavoidable condition during free flap transplantation. Restoration of blood flow is usually associated with a profound inflammatory response. Cytokines and growth factors are the functional proteins which exert their specific influence on injury or repair during the healing period. Plasma concentrations of 18 cytokines and growth factor proteins (IL6, IL8, IP10, TNFα, MCP1, Fractalkine, GRO, bFGF, GMCSF, IFNg, MIP1a, VEGF, sCD40L, IL10, TGFα, IL1β, IL12P40, and TNFβ) have been analyzed with respect to I/R status during microsurgery tissue transplantation in both, artery and vein, from patients by multiplexed immunoassay. Both technical feasibility and biostatistics data analysis approaches were thoroughly assessed. It has been found that, from all investigated proteins, the venous plasma levels of IL6 significantly increased during the ischemia period and mostly sustained their high levels during reperfusion, while venous plasma levels of IL8 showed in general a significant increase in the ischemia period followed by a rapid decrease in the reperfusion period. In conclusion, these findings direct toward an active involvement of tissue-resting leukocytes which may become therapeutic targets for concomitant medication in flap surgery to improve wound healing

Whole-scalp EEG mapping of somatosensory evoked potentials in macaque monkeys

High-density scalp EEG recordings are widely used to study whole-brain neuronal networks in humans non-invasively. Here, we validate EEG mapping of somatosensory evoked potentials (SSEPs) in macaque monkeys (Macaca fascicularis) for the long-term investigation of large-scale neuronal networks and their reorganisation after lesions requiring a craniotomy. SSEPs were acquired from 33 scalp electrodes in five adult anaesthetized animals after electrical median or tibial nerve stimulation. SSEP scalp potential maps were identified by cluster analysis and identified in individual recordings. A distributed, linear inverse solution was used to estimate the intracortical sources of the scalp potentials. SSEPs were characterised by a sequence of components with unique scalp topographies. Source analysis confirmed that median nerve SSEP component maps were in accordance with the somatotopic organisation of the sensorimotor cortex. Most importantly, SSEP recordings were stable both intra- and interindividually. We aim to apply this method to the study of recovery and reorganisation of large-scale neuronal networks following a focal cortical lesion requiring a craniotomy. As a prerequisite, the present study demonstrated that a 300-mm2 unilateral craniotomy over the sensorimotor cortex necessary to induce a cortical lesion, followed by bone flap repositioning, suture and gap plugging with calcium phosphate cement, did not induce major distortions of the SSEPs. In conclusion, SSEPs can be successfully and reproducibly recorded from high-density EEG caps in macaque monkeys before and after a craniotomy, opening new possibilities for the long-term follow-up of the cortical reorganisation of large-scale networks in macaque monkeys after a cortical lesion

Long-term follow-up of donor site morbidity after defect coverage with Integra following radial forearm flap elevation

BACKGROUND The radial forearm flap (RFF) is known for its excellent and reliable results regarding defect coverage though donor site morbidity represents a persisting problem. Integra is widely used in reconstructive surgery. This study aims to assess long-term outcomes regarding functionality and cosmesis after donor site coverage with respect to esthetic appearance, scar quality, and wrist function as well as ability to work after donor site coverage with Integra and split-thickness skin. METHODS The prospective follow-up exam, after a mean time of 23.8 months, enrolled 13 patients. Identification of patients' comorbidities and evaluation of the subjective esthetic outcome, sensibility, cold intolerance, and scar instability was assessed by a questionnaire. In clinics, designed Pinch test assessed scar tissue mobility over the flexor tendons. The Vancouver Scar Scale (VSS) was calculated; wrist function and grip strength were determined. RESULTS The satisfaction score for the esthetic appearance of the donor site was in average 3.42 ± 0.44. Two patients experienced an unstable scar and cold intolerance. The VSS resulted in a value of 4.2 representing a good result. The pinch test revealed an average scar mobility of 8 mm without any tendon adhesions. Active range of motion of the wrist was equal to the non-operated site as was grip strength. The pinch test showed a significant negative correlation with the VSS making it a reliable tool to measure scar quality. CONCLUSION Long-term results show an esthetic and functional successful defect coverage of the RFF donor site by the use of Integra and split-thickness skin. Level of Evidence: Level IV, therapeutic study

Mass spectrometric characterization of limited proteolysis activity in human plasma samples under mild acidic conditions

We developed a limited proteolysis assay for estimating dynamics in plasma-borne protease activities using MALDI ToF MS analysis as readout. A highly specific limited proteolysis activity was elicited in human plasma by shifting the pH to 6. Mass spectrometry showed that two singly charged ion signals at m/z 2753.44 and m/z 2937.56 significantly increased in abundance under mild acidic conditions as a function of incubation time. For proving that a provoked proteolytic activity in mild acidic solution caused the appearance of the observed peptides, control measurements were performed (i) with pepstatin as protease inhibitor, (ii) with heat-denatured samples, (iii) at pH 1.7, and (iv) at pH 7.5. Mass spectrometric fragmentation analysis showed that the observed peptides encompass the amino acid sequences 1-24 and 1-26 from the N-terminus of human serum albumin. Investigations on peptidase specificities suggest that the two best candidates for the observed serum albumin cleavages are cathepsin D and E. Reproducibility, robustness, and sensitivity prove the potential of the developed limited proteolysis assay to become of clinical importance for estimating dynamics of plasma-borne proteases with respect to associated pathophysiological tissue conditions