Different Cannabis sativa Extraction Methods Result in Different Biological Activities against a Colon Cancer Cell Line and Healthy Colon Cells.

Cannabis sativa is one of the oldest medicinal plants used by humans, containing hundreds of bioactive compounds. The biological effects and interplay of these compounds are far from fully understood, although the plant's therapeutic effects are beyond doubt. Extraction methods for these compounds are becoming an integral part of modern Cannabis-based medicine. Still, little is known about how different methods affect the final composition of Cannabis extracts and thus, their therapeutic effects. In this study, different extraction methods were tested, namely maceration, Soxhlet, ultrasound-assisted extraction (UAE), and supercritical CO2 extraction methods. The obtained extracts were evaluated for their cannabinoid content, antioxidant properties, and in vitro bioactivity on human colon cancer and healthy colon cells. Our data suggest that Cannabis extracts, when properly prepared, can significantly decrease cancer cell viability while protecting healthy cells from cytotoxic effects. However, post-processing of extracts poses a significant limitation in predicting therapeutic response based on the composition of the crude extract, as it affects not only the actual amounts of the respective cannabinoids but also their relative ratio to the primary extracts. These effects must be carefully considered in the future preparations of new therapeutic extracts.

Soft tissue grafts for dural reconstruction after meningioma surgery.

The meninges are involved in various pathologies and are often directly or indirectly severed during surgical procedures, especially the dura mater. This can pose a real challenge for the surgeon, as a proper reconstruction of the meninges is important to prevent complications such as cerebrospinal fluid leak (CSF). A variety of techniques for dural reconstruction have been described, employing natural and artificial materials. A novel technique for dural reconstruction involves soft tissue grafts in the form of fibrous or fibromuscular flaps, which are placed on the dural defects to seal the gaps. These soft tissue grafts represent an appropriate scaffold for cell ingrowth and fibrosis, thus preventing CSF. In this pilot study, we described the application of soft tissue grafts for dural reconstruction in 10 patients who underwent convexity meningioma surgery.

Systematic Evaluation of a Diclofenac-Loaded Carboxymethyl Cellulose-Based Wound Dressing and Its Release Performance with Changing pH and Temperature.

Development of drug-loaded wound dressings is often performed without systematic consideration of the changing wound environment that can influence such materials' performance. Among the crucial changes are the wound pH and temperature, which have an immense effect on the drug release. Detailed release studies based on the consideration of these changing properties provide an important aspect of the in vitro performance testing of novel wound dressing materials. A sodium carboxymethyl cellulose-based wound dressing, with the incorporated non-steroidal anti-inflammatory drug diclofenac, was developed and characterised in regard to its physico-chemical, structural and morphological properties. Further, the influence of pH and temperature were studied on the drug release. Finally, the biocompatibility of the wound dressing towards human skin cells was tested. Incorporation of diclofenac did not alter important properties (water retention value, air permeability) of the host material. Changes in the pH and temperature were shown to influence the release performance and have to be accounted for in the evaluation of such dressings. Furthermore, the knowledge about the potential changes of these parameters in the wound bed could be used potentially to predict, and potentially even to control the drug release from the developed wound dressing. The prepared wound dressing was also proven biocompatible towards human skin cells, making it interesting for potential future use in the clinics.

Novel ethanol-induced pectin-xanthan aerogel coatings for orthopedic applications.

In this study, we developed a novel high methoxyl pectin-xanthan aerogel coating on medical-grade stainless steel, prepared by ethanol-induced gelation and subsequent supercritical drying. Two non-steroidal anti-inflammatory drugs, i.e. diclofenac sodium and indomethacin, were incorporated into the aerogel coating. Electrochemical analyses were performed on the coated samples using electrochemical impedance spectroscopy and cyclic polarization techniques. The results showed that all passivated samples were highly resistant to general corrosion. The release of both non-steroidal anti-inflammatory drugs was complete after 24h, as confirmed by the plateau in the drug release profiles as well as by IR spectroscopy after the final release point. The potential of samples for use in orthopedic applications was evaluated on a human bone-derived osteoblast cell and all samples were shown to be biocompatible. The increased viability of some samples indicates the high potential of the developed approach for future evaluation of possible clinical use.

The influence of royal jelly and human interferon-alpha (HuIFN-αN3) on proliferation, glutathione level and lipid peroxidation in human colorectal adenocarcinoma cells in vitro.

Among royal jelly's (RJ) various biological activities, its possible antitumour activity deserves particular attention. The purpose of this study was to investigate the influence of RJ, its bioactive component 10-hydroxy-2-decenoic acid (10- HDA), and human interferon-alpha (HuIFN-αN3) on the proliferation of human colorectal adenocarcinoma cells (CaCo- 2), and ascertain their effect on intracellular glutathione (GSH) level and lipid peroxidation. We studied the antiproliferative (AP) activity of RJ [(0.1 g/10 mL phosphate buffer saline (PBS)], HuIFN-αN3 (1000 I.U. mL⁻¹), 10-HDA at 100.0 µmol L⁻¹, and their different combinations, in the ratio 1:1, 1:2, and 2:1 on CaCo-2 cells. The GSH level was measured by glutathione assay. The lipid peroxidation was measured by malondialdehyde (MDA) assay. Single RJ had a low AP activity: 2.0 (0.5 mg mL⁻¹). HuIFN-αN3 had an AP activity of 2.5 (208.33 I.U. mL⁻¹), while 10-HDA had an AP activity of 1.5 (37.5 µmol mL⁻¹). The highest AP activity of 3.8 was obtained when RJ and HuIFN-αN3 were applied at the ratio 2:1. In that combination the level of GSH was 24.9±2.4 nmol g⁻³ of proteins (vs. 70.2±3.2 nmol g⁻³ in the control) and the level of MDA was 72.3±3.1 nmol g⁻³ (vs. 23.6±9.1 nmol g⁻³ in the control). It is generally assumed that 10-HDA, an important constituent of RJ, together with HuIFN-αN3, is responsible for the inhibition of CaCo-2 cells proliferation in vitro. In our study, however, RJ and HuIFN-αN3 applied at 2:1 decreased the level of GSH the most and significantly increased lipid peroxidation via MDA in CaCo-2 cells. Future studies should show whether these GSH- and MDA-related activities of RJ, HuIFN-αN3, 10-HDA, and their combinations may decrease the tumorigenicity index and tumorigenic potential of various tumour cells in vitro.

Myogenic progenitors and imaging single-cell flow analysis: a model to study commitment of adult muscle stem cells.

Research on skeletal muscles suffers from a lack of appropriate human models to study muscle formation and regeneration on the regulatory level of single cells. This hampers both basic understanding and the development of new therapeutic approaches. The use of imaging multicolour flow cytometry and myogenic stem cells can help fill this void by allowing researchers to visualize and quantify the reaction of individual cultured cells to bioactives or other physiological impulses. As proof of concept, we subjected human CD56+ satellite cells to reference bioactives follistatin and Malva sylvestris extracts and then used imaging multicolor flow cytometry to visualize the stepwise activation of myogenic factors MyoD and myogenin in individual cells. This approach enabled us to evaluate the potency of these bioactives to stimulate muscle commitment. To validate this method, we used multi-photon confocal microscopy to confirm the potential of bioactives to stimulate muscle differentiation and expression of desmin. Imaging multicolor flow cytometry revealed statistically significant differences between treated and untreated groups of myogenic progenitors and we propose the utilization of this concept as an integral part of future muscle research strategies.