Fatty acid profiles in various lipid fractions in the female epidermis. Does the body site and age matter?

BACKGROUND: The epidermis forms the barrier between an organism and its external environment. Although one of the major functional elements of the epidermis is the lipid-enriched extracellular matrix, containing mainly ceramides, cholesterol (CHOL) and free fatty acids, the data are limited regarding the lipid profile in the epidermis. The aim of the study was to determine the whole profile of fatty acids (FAs) in the epidermis and to examine any dependence according to the age of the subject and the site on the epidermis. MATERIALS AND METHODS: Epidermis extracts obtained from 10 adults and 6 children were analyzed by gas chromatography-mass spectrometry. RESULTS: In total, 74 FAs in the human epidermis were identified. We observed the highest amounts of neutral lipids (including CHOL) compared to other lipid fractions in the epidermis, regardless of age. However, we detected an age-dependent content of the major lipid fractions, where the main difference was in the levels of polyunsaturated fatty acids. There were also differences in the lipid profile between various sites of the body, e.g. samples from the breast and abdomen were enriched with very long-chain fatty acids compared to the limb. CONCLUSION: Our research provides novel data concerning the lipid profile in the epidermis, gives further insight into skin biology and proves that the epidermis is a highly dynamic structure.

Efficiency assessment of irrigation as an alternative method for improving the regenerative potential of non-healing wounds.

The application of mesenchymal stem/stromal cells (MSC) in regenerative medicine offers hope for the effective treatment of incurable or difficult-to-heal diseases. However, it requires the development of unified protocols for both safe and efficient cell acquisition and clinical usage. The therapeutic effect of fat grafts (containing stem cells) in non-healing wounds has been discussed in previous studies, although the application requires local or general anaesthesia. The treatment of MSC derived from adipose tissue (ASC) could be a less invasive method, and efficient delivery could lead to more favourable outcomes, which should encourage clinicians to use such therapeutic approaches more frequently. Therefore, the aim of this study was to optimise the methods of ASC isolation, culture and administration while maintaining their high survival, proliferation and colonisation potential. The ASC were isolated by an enzymatic method and were characterised according to International Society for Cellular Therapy and International Federation for Adipose Therapeutics and Science guidelines. To assess the opportunity to obtain a sufficient number of cells for transplantation, long-term cell cultures in two oxygen concentrations (5% vs. 21%) were conducted. For these cultures, the population doubling time, the cumulative time for cell population doublings and the rate of cell senescence were estimated. In a developed and pre-defined protocol, ASC can be efficiently cultured at physiological oxygen concentrations (5%), which leads to faster proliferation and slower cell senescence. Subsequently, to select the optimal and minimally invasive methods of ASC transplantation, direct cell application with an irrigator or with skin dressings was analysed. Our results confirmed that both the presented methods of cell application allow for the safe delivery of isolated ASC into wounds without losing their vitality. Cells propagated in the described conditions and applied in non-invasive cell application (with an irrigation system and dressings) to treat chronic wounds can be a potential alternative or supplement to more invasive clinical approaches.

Secondary release of the peripheral nerve with autologous fat derivates benefits for functional and sensory recovery.

The reconstruction of nerve continuity after traumatic nerve injury is the gold standard in hand surgery. Immediate, tension-free, end-to-end nerve suture ensures the best prognosis. The recovery is mostly promising; however, in a few cases, insufficient outcomes in motor or sensory function are observed. Intra- and extra-fascicular scarring accompanies the nerve regeneration process and limits final outcomes. Secondary nerve release in those cases is recommended. Unfortunately, scarring recurrence cannot be eliminated after secondary revision and neurolysis. The supportive influences of mesenchymal stem cells in the process of nerve regeneration were observed in many preclinical studies. However, a limited number of studies in humans have analyzed the clinical usage of mesenchymal stem cells in peripheral nerve reconstruction and revisions. The objective of this study was to evaluate the effects of undifferentiated adipose-derived stromal/stem cell injection during a last-chance surgery (neurolysis, nerve release) on a previously reconstructed nerve. Three patients (one female, two males; mean age 59 ± 4.5 years at the time of injury), who experienced failure of reconstructions of median and ulnar nerves, were included in this study. During the revision surgery, nerve fascicles were released, and adipose-derived stromal/stem cells were administered through microinjections along the fascicles and around the adjacent tissues after external neurolysis. During 36 months of follow-up, patients noticed gradual signs of sensory and in consequence functional recovery. No adverse effects were observed. Simultaneous nerve release with adipose-derived stromal/stem cells support is a promising method in patients who need secondary nerve release after nerve reconstruction. This method can constitute an alternative procedure in patients experiencing recovery failure and allow improvement in cases of limited nerve regeneration. The study protocol was approved by the Institutional Review Board (IRB) at the Centre of Postgraduate Medical Education (No. 62/PB/2016) on September 14, 2016.

Shear Wave Elastography in the Evaluation of Temporomandibular Joint Disorders.

We aimed at verifying the usefulness of shear wave elastography in determining the temporomandibular disc stiffness in patients with a temporomandibular disorders (TMDs). The study included 37 patients with confirmed TMDs and 208 healthy volunteers. Patients presented with significantly greater stiffness of the intermediate zone of the disc (region of interest [ROI] 1) and significantly lower stiffness of its anteriorly displaced portion (ROI 3). A receiver operating characteristics analysis indicated that a decrease in the stiffness in ROI 3 less than 8.667 KPa provided 100% sensitivity, 97.3% specificity, 100% positive predictive value (PPV) and 99.5% negative predictive value (NPV) in distinguishing between patients with TMDs and without. Whereas an increase in ROI 1 stiffness to at least 54.33 KPa provided high specificity and NPV, both the sensitivity and the PPV of this predictor equaled zero. Findings suggest that a decrease in anteriorly dislocated disc stiffness less than 8.667 kPa can accurately identify patients with TMDs.

Intraspinal Transplantation of the Adipose Tissue-Derived Regenerative Cells in Amyotrophic Lateral Sclerosis in Accordance with the Current Experts' Recommendations: Choosing Optimal Monitoring Tools.

Stem cells (SCs) may constitute a perspective alternative to pharmacological treatment in neurodegenerative diseases. Although the safety of SC transplantation has been widely shown, their clinical efficiency in amyotrophic lateral sclerosis (ALS) is still to be proved. It is not only due to a limited number of studies, small treatment groups, and fast but nonlinear disease progression but also due to lack of objective methods able to show subtle clinical changes. Preliminary guidelines for cell therapy have recently been proposed by a group of ALS experts. They combine clinical, neurophysiological, and functional assessment together with monitoring of the cytokine level. Here, we describe a pilot study on transplantation of autologous adipose-derived regenerative cells (ADRC) into the spinal cord of the patients with ALS and monitoring of the results in accordance with the current recommendations. To show early and/or subtle changes within the muscles of interest, a wide range of clinical and functional tests were used and compared in order to choose the most sensitive and optimal set. Additionally, an analysis of transplanted ADRC was provided to develop standards ensuring the derivation and verification of adequate quality of transplanted cells and to correlate ADRC properties with clinical outcome.

Autologous Fat Transfer in Secondary Carpal Tunnel Release.

BACKGROUND: Carpal tunnel release is the gold standard for the treatment of median nerve compression disease. Recurrent or persistent symptoms do not occur in most patients, although a small number of them have indicated that such a postoperative condition indeed exists. Some patients undergo repeated treatments. In the majority of the cases, the disease is associated with scarring in the carpal tunnel or even reformation of the carpal ligament. The authors propose the usage of autologous fat grafting during secondary carpal tunnel release to inhibit the scarring process. METHODS: Ten patients with recurrent or persistent symptoms underwent autologous fat grafting at the time of their repeated carpal tunnel release. Fat was harvested from the lower abdomen and grafted into the scarred transverse carpal ligament and surrounding tissues. Each patient underwent pre- and postoperative examinations and completed the carpal tunnel questionnaire (Boston) to evaluate their sensory and motor functions. The patients underwent 1 year of follow-up. RESULTS: There were 2 main reasons for continued symptoms: a technical mistake resulting in incomplete release (IR) during the first operation and abundant scarring (ABS) in the operated area. The beneficial effects of the interventions were confirmed by a clinical study and by administering the carpal tunnel questionnaire to all patients (functional severity score decreased from 4.38 to 1.88 in IR and 3.62 to 1.48 in ABS group, sensory severity score from 3.26 to 1.7 in IR and 3.04 to 1.48 in ABS group; P < 0.05) after 12 months of follow-up. CONCLUSION: Our initial observations suggest the possible efficacy of adipose tissue in secondary carpal tunnel release.

Feasibility and limitations of endoscopy in Guyon's canal.

INTRODUCTION: This retrospective report summarizes observations from eight operations where the endoscopically assisted approach was used to explore Guyon's canal syndromes of idiopathic aetiology. AIM: To evaluate the feasibility and limitations of endoscopic Guyon's canal release performed from a distal forearm incision. MATERIAL AND METHODS: Eight charts and video records of eight ulnar tunnel syndrome patients presenting concomitant idiopathic Guyon's canal syndromes were retrospectively reviewed. In all cases endoscopically assisted explorations in Guyon's canals with simultaneous cubital tunnel releases were performed. RESULTS: In all of the patients the multiple tight bands of the superficial volar carpal ligament forming the canal roof were divided. Some of these bands crossing the nerve in its direct vicinity could have been responsible for the constriction. We were also able to divide the proximal segment of the canal floor. We have observed, however, that the proximal to distal endoscopic dissection jeopardizes the motor branch of the ulnar nerve; therefore, it should not be used to release the pisohamate ligament, or the hypothenar fascia. CONCLUSIONS: Although all of the patients showed improvement, we cannot recommend this method in its current form. We are of the opinion that safe endoscopic Guyon's canal operations may require a different approach.

Superficial plane endoscopy for carpal tunnel release.

In this publication we describe three techniques for endoscopic carpal tunnel release which use the palmar portal approach and the superficial plane to enable a view from above the transverse carpal ligament. These techniques prevent any iatrogenic compression of the median nerve inside the already narrow tunnel. Although the necessary instruments are inexpensive and recommended in instrumentation catalogues, their use has rarely been reported. These techniques are safe and easy to learn for surgeons familiar with arthroscopy. We believe that they constitute an important alternative to the already well-established methods of interior tunnel access, and believe that their use should become more widespread.

Integrin expression in the dermis during scar formation in humans.

To evaluate changes leading to human wound reorganization we examined by immunohistochemistry the expression of several extracellular matrix (ECM) receptors (alpha2 chain of VLA-2, alpha3 chain of VLA-3, alpha6 chain of VLA-6, alphav, and beta1/beta3 chains of integrins) in a series of biopsies of human skin wounds healing by primary intention. The first time point investigated in this study was day 6 after injury, i.e. when a fibrin clot has been almost completely replaced by the granulation tissue. Gradual changes in integrin expression in granulation tissue and in the dermal scar were observed from the first time point investigated and were characterized by an up-regulation of alpha2beta1 complex, alphav integrin subunit, and beta1 integrin subunit. At day 27, the expression of the alpha2 chain of VLA-2 in the scar decreased. The expression of alphav and beta1 integrin subunits decreased but was still detectable by day 35. Vitronectin expression from day 7 onwards was also increased and colocalized to the area of the wounded dermis, and decreased by day 27. Our data suggests that, during the remodelling of the provisional matrix of the wound, dermal fibroblasts express transiently mainly alpha2 and alphav subunits of integrins associated with up-regulation of the beta1 subunit. It seems that up-regulation of some chains of integrins may be involved in the control of deposition of ECM components associated with wound healing.