Anna Ścisłowska-Czarnecka

Low-level laser therapy (LLLT) is being increasingly applied in regenerative medicine due to its ability to modulate cellular metabolism and support repair processes. However, the effect of various exposure parameters on fibroblast activity still does not remain fully understood. The study aim was to evaluate the effect of selected LLLT parameters (808 nm, 100 and 200 mW, 2 and 10 J/cm², pulsed beam) on the biological activity of fibroblasts in vitro. Fibroblasts were exposed to the established exposure parameters, and then the number of adherent and metabolically active cells, their migration, adenylate kinase (AK) as well as nitric oxide (NO) levels, cytokine secretion, metalloproteinase (MMP-2, MMP-9) activity, and oxidative-antioxidant potential were assessed. The most favourable effects were achieved with 200 mW; 2 J/cm² and 100 m; 10 J/cm² exposure parameters. In these conditions, increased number of adherent and metabolically active cells, and elevated migration were observed. Moreover, in this condition decreased AK release and NO secretion were found. Furthermore, LLLT reduced in fibroblasts interleukin 6 (IL-6) and interleukin 8 (IL-8) secretion as well as affected the activity of metalloproteinases, what corresponded with subsequent wound healing process. Additionally, a beneficial effect on maintaining the oxidative-antioxidant balance in fibroblasts was noted. In summary, the effects of LLLT on fibroblasts depend on the applied irradiation parameters, and appropriately selected LLLT treatment (200 mW; 2 J/cm² and 100 mW; 10 J/cm²,) stabilise fibroblast metabolism, limits their proinflammatory response and promotes extracellular matrix remodelling (ECM). These results confirm the potential of LLLT as an effective method supporting regenerative processes in the treatment of tissue damage.

Modulation of macrophage function represents a promising strategy for promoting tissue remodelling and repair. In the present study the combined effect of low-level laser therapy (LLLT) and α-lipoic acid (LA) on resting RAW 264.7 macrophages cultured in vitro was evaluated. Cells were cultured on LA-modified poly(ε-caprolactone) (PCL) membranes, designed as a controlled release platform. Physicochemical analysis confirmed that LA incorporation decreased PCL crystallinity while increasing membrane surface porosity, providing a biphasic LA release profile. LLLT (808 nm, 200 mW, 5 J/cm², pulsed mode) did not affect macrophage viability but modulated matrix metalloproteinase (MMP-9 and 2) activity and macrophage oxidative-antioxidant potential. Cell culturing carried out on LA-modified membranes (PCL-LA) increased their viability, reduced AK and NO levels as well as TNF-α secretion. Simultaneous treatment with LLLT and LA (PCL-LA-IR) further increased macrophage viability, decreased level of MCP-1/CCL2 chemokine, and modified the presence of pro-MMP-9 as well as the activity of MMP-2, MMP-9 and MMP-9 dimers. Moreover, LA-treated cells showed an increase in antioxidant capacity without oxidative potential being altered. These results indicate that PCL is a stable and effective carrier of LA, and that the observed combined effect of LLLT and LA influences the activity of resting macrophages, which may promote the creation of a favourable environment for regenerative processes. These results also indicate the potential for combined therapies using immunomodulatory biomaterials that support tissue repair to be further developed.

Tissue adhesives offer a promising alternative to traditional sutures and staples, particularly in situations requiring rapid, minimally invasive wound closure. To address the limitations of commercially available cyanoacrylate-based adhesives, numerous hydrogel adhesives have been developed. This study presents the synthesis and characterisation of an alginate–aminophenylboronic acid (Alg-APBA) hydrogel adhesive, optimised for bioprinting as a method allowing us to control the thickness of the adhesive layer. The adhesive combines the biocompatibility of alginate with the pH-responsive bonding ability of boronic acid groups, eliminating the need for oxidative crosslinkers. Successful conjugation of APBA to alginate was confirmed via ¹H NMR, FTIR and UV-VIS spectroscopy, with a degree of substitution reaching approximately 46% or ~0.22 mol%. Rheological analysis demonstrated shear-thinning and self-healing properties suitable for bioprinting, achieving a high print fidelity (Pr ratio = 0.99 ± 0.08) and repeatability. Mechanical testing showed a shear strength of 19.0 ± 0.5 kPa and an interfacial toughness of 58.0 ± 2.11 J/m², exceeding those of commercial fibrin adhesives. Additionally, the adhesive joint remained stable after one week of incubation in an acidic environment. The material demonstrated biocompatibility during in vitro testing with keratinocytes and fibroblast cells. These results indicate that Alg-APBA is a strong, biocompatible and printable hydrogel adhesive with potential applications in soft tissue implant fixation.

Cel pracy: Celem pracy było opisanie zmian w stabilności posturalnej, jakości życia, funkcjach poznawczych i zachowaniu pacjentów chorych na schizofrenię paranoidalną, którzy przeszli terapię Snoezelen i program ćwiczeń sensomotorycznych.Metoda: Badanie przeprowadzono wśród 55 pacjentów w wieku od 55 do 75 lat. Grupa S była poddana terapii w sali Snoezelen przez dziewięć tygodni, grupa ES została poddana terapii Snoezelen i została również poddana programowi ćwiczeń sensomotorycznych, a grupa C nie otrzymała żadnej interwencji terapeutycznej. Do oceny efektów terapii wykorzystano przenośną platformę stabilometryczną Biosway, Berg Balance Scale, Tinetti Test oraz Timed Up & Go Test.Wyniki: W grupie ES zaobserwowano istotne statystycznie zmiany wszystkich badanych wskaźników; w grupie S zmiany takie zaobserwowano w teście Granic Stabilności, teście Tinnettiego i teście TUG; natomiast w grupie C nie stwierdzono zmian istotnych statystycznie.Wnioski: Terapia Snoezelen istotnie wpłynęła na zmiany w zakresie kontroli postawy i zmniejszenia ryzyka upadków. Autorski program ćwiczeń sensomotorycznych zwiększył siłę oddziaływania sali Snoezelen na stabilność posturalną mierzoną za pomocą platformy Biosway oraz ryzyko upadków oceniane za pomocą testu TUG.

Background/Objectives: Low-level laser therapy (LLLT) covers a wide range of parameters in terms of laser properties and dosage, which is important for its effects. It is important to select safe, optimal irradiation conditions to obtain the desired therapeutic effect of LLLT on cells. This article is focused on the selection of favourable (biostimulating) exposure conditions for LLLT, which are the beam application method (continuous [C] or pulsed [P] laser beam), radiation power and LLLT dose, on the viability and secretory activity regarding resting macrophages of the RAW 264.7 cell line. Methods: RAW 264.7 macrophages were seeded on 24-well tissue culture. ASTAR PhysioGo 400C apparatus with a spot applicator generating electromagnetic radiation in the infrared light range of 808 nm and power of 100 mW and 200 mW was used for laser irradiation of macrophages. Cells were treated with different doses of constant radiation 5 J/cm²/well or 10 J/cm²/well. Results: It was shown that the most beneficial radiation parameters for cells were obtained with a pulsed laser beam of 200 mW power and a dose of 5 J/cm², which caused an increase in macrophage adhesion and viability, as well as an increase in NO secretion by macrophages and their TOS, with a simultaneous decrease in the secretion of TNF-α, MCP-1 and MMP-9 by cells. Conclusions: The research results presented above indicate that the effect of LLLT on resting macrophages modulates their biological activity, and the intensity of photobiostimulation depends on the irradiation parameters, including wavelength, power, dose and method of laser beam application.

For several decades, there has been growing interest in the influence of low-frequency magnetic fields (LFMFs) and red LED light on the healing process. Keratinocytes are cells that play a significant role in the process of wound healing and tissue regeneration. A human keratinocyte cell line (HaCaT) was exposed to an LFMF with low induction (180–195 Hz; 60 µT, magnetostimulation), red LED light (630 nm; 300 mW, LED therapy), and their combined action (magneto-LED therapy) in in vitro culture conditions. On day 4 and 8 of the experiment, the following parameters were determined: adhesion/proliferation, adenylate kinase (AK), nitric oxide (NO), cytokines (IL-1β, IL-6, IL-8, IL-10, IL-12p70, TNF-α), metalloproteinases (MMP-2, MMP-9), and collagen IV. It was shown that magnetostimulation caused an increase in keratinocyte adhesion/proliferation and IL-8 secretion and a decrease in IL-12 secretion. The LED therapy resulted in a transient increase in the secretion of NO and cytokines IL-1, IL-12, and IL-6 in keratinocytes. The use of magneto-LED therapy resulted in an increase in keratinocyte adhesion/proliferation, the secretion of cytokines IL-6 and IL-8, and NO with a simultaneous decrease in MMP-9 secretion. The results of our studies showed that the action of an LFMF with low-induction and LED light on keratinocytes can modulate the biological activity of keratinocytes towards improving the skin healing process.

Emulsion electrospinning is a method of modifying a fibers’ surface and functional properties by encapsulation of the bioactive molecules. In our studies, bovine serum albumin (BSA) played the role of the modifier, and to protect the protein during the electrospinning process, the W/O (water-in-oil) emulsions were prepared, consisting of polymer and micelles formed from BSA and anionic (sodium dodecyl sulfate–S) or nonionic (Tween 80–T) surfactant. It was found that the micelle size distribution was strongly dependent on the nature and the amount of the surfactant, indicating that a higher concentration of the surfactant results in a higher tendency to form smaller micelles (4–9 µm for S and 8–13 µm for T). The appearance of anionic surfactant micelles reduced the diameter of the fiber (100–700 nm) and the wettability of the nonwoven surface (up to 77°) compared to un-modified PCL polymer fibers (100–900 nm and 130°). The use of a non-ionic surfactant resulted in better loading efficiency of micelles with albumin (about 90%), lower wettability of the nonwoven fabric (about 25°) and the formation of larger fibers (100–1100 nm). X-ray photoelectron spectroscopy (XPS) was used to detect the presence of the protein, and UV-Vis spectrophotometry was used to determine the loading efficiency and the nature of the release. The results showed that the location of the micelles influenced the release profiles of the protein, and the materials modified with micelles with the nonionic surfactant showed no burst release. The release kinetics was characteristic of the zero-order release model compared to anionic surfactants. The selected surfactant concentrations did not adversely affect the biological properties of fibrous substrates, such as high viability and low cytotoxicity of RAW macrophages 264.7.

Seniors are a constantly growing group of people in many societies. It is necessary to develop physiotherapeutic programs to improve their mobility. The aim of this study was to assess the impact of the physiotherapeutic program conducted unstable ground on selected indicators of motor functions of elderly women. Sixty women (60–80 years) participated in the research. Group A (N = 20) underwent a 12-week physiotherapeutic program on stable ground, group B (N = 20) followed an exercise program on unstable ground, and group C (N = 20) (control group) had no therapeutic intervention. The effects of the therapy were assessed by using a FreeMed platform (foot load analysis) and a Biosway balance system. The results were compared using ANOVA (the one-way analysis), the Kruskal–Wallis test and also the post hoc tests (Tukey’s test and the multiple comparison test). In group A, a statistically significant change was observed in the static test and balance assessment, in group B this was observed in the static and dynamic foot tests and balance assessment, in group C, no statistical significance was achieved. The authors’ physiotherapeutic program had a statistically significant effect on changes in the balance and selected indicators of the motor functions of the examined people. Comparing the results before and after the therapy more improvement changes were noted in women training on an unstable ground compared to women training on a stable ground.

This study assessed the effect of Pilates exercises on balance and fall risk in older women. Participants comprised 50 older women aged over 60 years, divided randomly into two groups: the experimental group (n = 30), which took part in Pilates sessions two times per week for three months, and the control group (n = 20). The control group did not participate in such sessions but also did not participate in any other rehabilitation programs or additional physical activity except everyday activities. Before and after the training cycle, all women underwent an assessment using Timed Up and Go (TUG), the One Leg Stance Test (OLST), a test performed on a Freestep baropodometric platform, and the tests performed on a Biosway platform. After the training, significantly decreased values of the surface of the ellipse (p = 0.0037) and mean values of velocity (p = 0.0262) for the right foot in the experimental group were observed. The Limits of Stability (LoS) test (p = 0.005) and the Modified Clinical Test of Sensory Interaction on Balance (m-CTSIB) performed on an unstable surface with eyes closed (p = 0.0409) indicated statistically significant changes in the experimental group. None of the above changes were statistically significant in relation to the control group. Pilates training affected the participants’ balance by improving LOS and reducing fall risk.

Skin regeneration requires a three-dimensional (3D) scaffold for cell adhesion, growth and proliferation. A type of the scaffold offering a 3D structure is a nonwoven material produced via a melt-blown technique. Process parameters of this technique can be adapted to improve the cellular response. Polylactic acid (PLA) was used to produce a nonwoven scaffold by a melt-blown technique. The key process parameters, i.e., the head and air temperature, were changed in the range from 180–270 °C to obtain eight different materials (MB1–MB8). The relationships between the process parameters, morphology, porosity, thermal properties and the cellular response were explored in this study. The mean fiber diameters ranged from 3 to 120 µm. The average material roughness values were between 47 and 160 µm, whereas the pore diameters ranged from 5 to 400 µm. The calorimetry thermograms revealed a correlation between the temperature parameters and crystallization. The response of keratinocytes and macrophages exhibited a higher cell viability on thicker fibers. The cell-scaffold interaction was observed via SEM after 7 days. This result proved that the features of melt-blown nonwoven scaffolds depended on the processing parameters, such as head temperature and air temperature. Thanks to examinations, the most suitable scaffolds for skin tissue regeneration were selected.