Background/Objectives: Efficient wound treatment embraces the management of four overlapping phases, starting with hemostasis, an immediate physiological response aimed at stopping bleeding from damaged blood vessels caused by skin injury. This paper proposes an innovative, nature-based hemostatic biomaterial designed to assist natural self-healing regenerative mechanisms. Methods: Light, transparent, and skin-adhesive films based on κ-carrageenan, meadow polyfloral honey, and Calendula officinalis flower extract were fabricated via solution casting. Comprehensive characterization revealed the physicochemical, structural, swelling, and barrier properties and the influence of each bioactive compound utilized for film preparation. Results: The samples subcutaneously implanted in Wistar rats induced vascularization, deposition of collagen, and orientation of collagen fibers while being fully phagocytosed and gradually biodegraded. The rat tail-cut model demonstrated that the films significantly reduced blood loss (0.1875 ± 0.0732 g) compared to the control (0.7837 ± 0.3319 g), and hemostasis was achieved notably faster (355.75 ± 71.42 s) than in the control group (704.25 ± 85.29 s). The rat liver punch biopsy model confirmed reduced blood loss (2.8025 ± 1.5174 g) and shorter time to hemostasis (303.25 ± 77.90 s) compared to the control (3.1475 ± 1.5413 g, 383.00 ± 36.53 s). Conclusions: The results indicate the great potential of the fabricated films as hemostatic wound dressings.

Scar formation and delayed wound healing pose significant challenges in treating skin injuries, especially in severe cases like burns and diabetic wounds. This study investigates the effectiveness of novel Poly(vinyl alcohol) (PVA)/Gentamicin (Gent) and PVA/Chitosan (CHI)/Gent hydrogels in promoting healing of second-degree burn wounds in a rat model. Following in vitro testing, these hydrogels were deemed non-toxic and suitable for in vivo analysis. Clinical evaluations were conducted on the 3rd, 7th, 14th, and 21st post-injury days, assessing parameters such as blistering, edema, redness, crust, bleeding, secretion, scar tissue formation, and wound contraction percentage. Histological analyses focused on re-epithelization and dermal evaluation at specific time points. Results showed that both hydrogels significantly reduced inflammation, particularly redness, by the 14th day and improved re-epithelization, with the PVA/CHI/Gent group outperforming on the 14th day and the PVA/Gent group excelling on the 21st day. Histological findings indicated increased fibroblast proliferation and collagen deposition in treated groups, suggesting enhanced dermal healing. The PVA/CHI/Gent hydrogel demonstrated notable antibacterial properties, likely due to the synergistic effects of CHI and Gent, leading to reduced inflammation and edema. Overall, both hydrogels show promise as effective wound dressings, facilitating faster healing and improved tissue recovery in burn injuries. This study supports the use of biomimetic scaffolds for enhanced wound management in clinical practices.

In this paper, we have shown for the fi rst time the soft tissue response of novel silver/ poly(vinyl alcohol)/graphene (Ag/PVA/Gr) and silver/poly(vinyl alcohol)/chitosan/ graphene (Ag/PVA/CHI/Gr) nanocomposite hydrogels aimed for medical applications. These novel hydrogels were produced by in situ electrochemical synthesis of silver nanoparticles in the polymer matrices as described in our previously published works. Both Ag/PVA/Gr and Ag/PVA/CHI/Gr, as well as controls Ag/PVA, Ag/PVA/CHI and commercial Suprasorb ^© hydrogel discs, were implanted in the subcutaneous tissue of rats. Implants with the surrounding tissue were dissected after post-implantation on days 7, 15, 30 and 60, and then processed for histological examination. The tissue irritation index (TIrI) score, according to ISO 10993-6, 2007, as well as the number of leukocytes in the peri-implant zone and connective tissue capsule thickness were examined. The results show that each TIrI score, the leukocyte number around the implanted materials and capsule thickness gradually decreased during the observation period. At the endpoint of follow-up, the Ag/PVA/CHI/Gr implant was surrounded with a thinner capsule, while both the TIrI score and the number of leukocytes of the peri-implant zone were greater compared to the Ag/PVA/Gr implant. Despite the observed differences, we can conclude that our in vivo experiment suggested that both novel hydrogels were biocompatible and suitable for medical use.

Background/Aim. The last decade has been profoundly marked by persistent attempts to use ex vivo expanded and manipulated mesenchymal stem cells (MSCs), as a tool in different types of regenerative therapy. In the present study we described immunophenotype and the proliferative and differentiation potential of cells isolated from pulp remnants of exfoliated deciduous teeth in the final phase of root resorption. Methods. The initial adherent cell population from five donors was obtained by the outgrowth method. Colony forming unit-fibroblast (CFU-F) assay was performed in passage one. Cell expansion was performed until passage three and all tests were done until passage eight. Cells were labeled for early mesenchymal stem cells markers and analysis have been done using flow cytometry. The proliferative potential was assessed by cell counting in defined time points and population doubling time was calculated. Commercial media were used to induce osteoblastic, chondrogenic and adipogenic differentiation. Cytology and histology methods were used for analysis of differentiated cell morphology and extracellular matrix characteristics. Results. According to immunophenotype analyses all undifferentiated cells were positive for the mesenchymal stem cell markers: CD29 and CD73. Some cells expressed CD146 and CD106. The hematopoietic cell marker, CD34, was not detected. In passage one, incidence of CFU-F was 4.7 ? 0.5/100. Population doubling time did not change significantly during cell subcultivation and was in average 25 h. After induction of differentiation, the multicolony derived cell population had a tri-lineage differentiation potential, since mineralized matrix, cartilage-like tissue and adipocytes were successfully formed after three weeks of incubation. Conclusion. Altogether, these data suggest that remnants of deciduous teeth dental pulp contained cell populations with mesenchymal stem cell-like features, with a high proliferation and trilineage differentiation potential and that these cultures are suitable for further in vitro evaluation of cell based therapies.

The ethical dilemmas of using animals as in vivo models in preclinical and clinical examinations have been increasingly present in recent decades. Small laboratory animals (rats, rabbits) will continue to be used because they are cost-effective and permit the formation of statistically testable cohort groups; a task that, for financial, maintenance and care reasons, is almost prohibitive for larger animals. Technological advances in the production of new biomaterials for clinical use are enormous, but screening tests and methods used to assess biocompatibility lag behind these advances. The assessment of biological responses is slow and based on millennial recovery mechanisms in eukaryotic organisms. Therefore, the goal of researchers in this field is to re-evaluate old methods of biocompatibility assessment and introduce new methods of evaluation, especially for in vivo testing. In that sense, a revision of the ISO standards was planned and conducted in 2017, which insisted on cytotoxicity testing in cell lines and produced concrete proposals on how biocompatibility should be quantified. In vivo biocompatibility evaluation of biomaterials used for soft tissue recovery commonly utilises rats. Rabbits are recommended for implants used for hard tissues, because of the rabbit?s size, the possibility of implanting the biomaterials on a larger bone surface, and because of the peculiarities of rabbit bone tissue that favours rapid recovery after bone defects and enables easy reading of the results