Biocompatible 3D-Printed Devices With Adipose Stem Cells in the Regenerative Process of Sciatic Nerve Lesions in Rodent Models: An Experimental Study.

INTRODUCTION: Peripheral nerve injuries are a significant clinical challenge. The rat sciatic nerve serves as an ideal model for studying nerve regeneration. Extensive research has been conducted to unravel the intricate mechanisms involved in peripheral nerve regeneration, aiming to develop effective therapeutic strategies for nerve injury patients. Research including different types of materials that can be used as nerve guides like synthetic polymers have been investigated for their biocompatibility and molding properties. Among multiple stem cell types, adipose-derived stem cells (ASCs), bone marrow-derived mesenchymal stem cells (BM-MSCs), and induced pluripotent stem cells (iPSCs) have shown neuroprotective and regenerative important properties. METHODS: The purposes of our study were to develop a protocol for rat sciatic nerve injury treated with 3D-printed guide and adipose stem cells to investigate nerve regeneration through histologic examination and biomechanical characteristics of muscular tissue. We use 20 (100%) male Wistar rats, measuring between 350 g ± 35 g, who underwent complete transection of the right sciatic nerve, resulting in a 1 cm defect. The group was separated into three subgroups: the first subgroup (n = 8) was treated with a 3D-printed guide with adipose stem cells, the second subgroup (n = 8) was treated with a 3D-printed guide without adipose stem cells, and the third subgroup (n = 4) was the control group. At four, eight, and 12 weeks, we measured with ultrasonography the grade of muscular atrophy. At 12 weeks, we harvested the sciatic nerve and performed a histological examination and mechanical investigation of the tibialis anterior muscle. RESULTS: On the examined specimen of the first subgroup, cross-sectioned nerve structures were present, surrounded by a mature fibro-adipose connective tissue, with blood vessels. In the second subgroup, no nerve structure was observed on the examined sections, but in the polymorphic inflammatory infiltrate and control group, no signs of regeneration were found. CONCLUSIONS: The present study shows a promising potential when utilizing adipose stem cell-based therapies for promoting peripheral nerve regeneration following large (>1 cm) nerve defects knowing that at this size, regeneration is impossible with known treatments.

Application of Adipose Stem Cells in 3D Nerve Guidance Conduit Prevents Muscle Atrophy and Improves Distal Muscle Compliance in a Peripheral Nerve Regeneration Model.

BACKGROUND: Peripheral nerve injuries (PNIs) represent a significant clinical problem, and standard approaches to nerve repair have limitations. Recent breakthroughs in 3D printing and stem cell technologies offer a promising solution for nerve regeneration. The main purpose of this study was to examine the biomechanical characteristics in muscle tissue distal to a nerve defect in a murine model of peripheral nerve regeneration from physiological stress to failure. METHODS: In this experimental study, we enrolled 18 Wistar rats in which we created a 10 mm sciatic nerve defect. Furthermore, we divided them into three groups as follows: in Group 1, we used 3D nerve guidance conduits (NGCs) and adipose stem cells (ASCs) in seven rats; in Group 2, we used only 3D NGCs for seven rats; and in Group 3, we created only the defect in four rats. We monitored the degree of atrophy at 4, 8, and 12 weeks by measuring the diameter of the tibialis anterior (TA) muscle. At the end of 12 weeks, we took the TA muscle and analyzed it uniaxially at 10% stretch until failure. RESULTS: In the group of animals with 3D NGCs and ASCs, we recorded the lowest degree of atrophy at 4 weeks, 8 weeks, and 12 weeks after nerve reconstruction. At 10% stretch, the control group had the highest Cauchy stress values compared to the 3D NGC group (0.164 MPa vs. 0.141 MPa, p = 0.007) and the 3D NGC + ASC group (0.164 MPa vs. 0.123 MPa, p = 0.007). In addition, we found that the control group (1.763 MPa) had the highest TA muscle stiffness, followed by the 3D NGC group (1.412 MPa), with the best muscle elasticity showing in the group in which we used 3D NGC + ASC (1.147 MPa). At failure, TA muscle samples from the 3D NGC + ASC group demonstrated better compliance and a higher degree of elasticity compared to the other two groups (p = 0.002 and p = 0.008). CONCLUSIONS: Our study demonstrates that the combination of 3D NGC and ASC increases the process of nerve regeneration and significantly improves the compliance and mechanical characteristics of muscle tissue distal to the injury site in a PNI murine model.

[Laparoscopic treatment of appendiceal mucocele]. / Appendix mucokele laparoscopes mûtéti megoldása.

Mucocele is a mucin-filled cavity, which can be multi-loculated as well. Mucocele is relatively rarely found in the appendix. The disease does not cause any symptoms in most cases, and it is usually an accidental finding. A thorough investigation should be carried out to exclude malignancy. Depending on the operative findings and the full histological report, the spectrum of surgery extends from appendectomy to right hemicolectomy. In this paper, we discuss the presentation, diagnostical options, and the possible surgical treatment--based on our experience with five cases. In three cases a non-tender mass was palpable in the right lower quadrant of the abdomen, while another patient presented with right lower quadrant abdominal pain and one with abdominal pain and diarrhoea. Abdominal and pelvic ultrasound and CT scans raised the possibility of mucocele, however colonoscopy was negative. After laparoscopic exploration, laparoscopy assisted partial caecum resection was carried out in two cases, in further two cases laparoscopic appendectomy, and in one case laparoscopic partial caecum resection was done. The hystological examinations showed appendiceal mucocele with no malignancy demonstrated. All patients recovered without complications, they were discharged from hospital on postoperative day five. The patients have been disease free after a 6-30 month follow-up period. We concluded that laparoscopy is a recommended method for the surgery of appendiceal mucocele.

High-performance liquid chromatographic-tandem mass spectrometric method for the determination of clemastine in human plasma.

A highly sensitive high-performance liquid chromatographic-tandem mass spectrometric method (HPLC-MS-MS) has been developed to quantitate clemastine in human plasma for the purpose of pharmacokinetic studies. Sample preparation was carried out by liquid-liquid extraction using deuterated clemastine as an internal standard. Chromatographic separation used a C18 reversed phase polymer column giving an extremely fast total run time of 2 min. The method was validated and used for the bioequivalence study of clemastine tablets in healthy male volunteers (n=28). The lower limit of detection proved to be 0.01 ng/ml for clemastine.

Characterization of the selectivity of a phenytoin imprinted polymer.

The selectivity of analytical methods based on molecularly imprinted polymers (MIPs) is due to the preferential adsorption of the analyte(s) as compared to other substances (interferences). This paper shows the theoretical and practical difficulties, which have to be considered and solved when real samples need to be analysed in a wide range of analyte and interferant concentrations. It is shown that the estimation of interference effects requires either many measurements or a realistic model of the adsorption equilibrium in mixed solutions of the analyte and the interferences. Examples are shown for positive (cooperative) interference effects, for better experimental design and interpretation of binary isotherm measurements and for establishing the chemical model of interference from selectivity measurements. The usual MIP model consisting of a cavity, which closely fits the shape of the template from all sides, appears unsuitable for this MIP, and it is replaced with a more realistic, more open model. The applicability of the results to using non-imprinted polymers as selective sorbents and to screening drug candidates is also shown.

Effect of solvents on the selectivity of terbutylazine imprinted polymer sorbents used in solid-phase extraction.

A solid-phase extraction sample preparation method using a molecularly imprinted polymer (MIP) selective for the triazine type pesticide terbutylazine has been developed. The method involves preconcentration from large volumes of water samples on a C18 disk coupled to selective clean-up on the MIP. The method has been optimised by studying the recovery and retention of terbutylazine and some other structurally related triazine derivates as a function of the selective washing solvent used. The effect of the water content of the selective washing solvent was also investigated on the recovery of the MIP. River water samples were analysed with the coupled technique, and efficient clean-up of the samples was observed.

Binding assays with molecularly imprinted polymers--why do they work?

The design of homologous displacement ligand binding assays based on molecularly imprinted polymers (MIP) is discussed in terms of the MIP adsorption isotherm. It is shown that only MIPs having a binding isotherm with varying slope are suitable for the assay, but there is no need to interpret the isotherm in terms of site affinity and population. One can calculate the calibration plot of the binding assay from the isotherm and vice versa.