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The mesenchymal stem cell (MSC) secretome has been considered an innovative therapeutic biological approach, able to modulate cellular crosstalk and functionality for enhanced tissue repair and regeneration. This study aims to evaluate the functionality of the secretome isolated from periosteum-derived MSCs, from either basal or osteogenic-induced conditions, in the healing of a critical size calvarial bone defect in the rabbit model. A bioceramic xenograft was used as the vehicle for secretome delivery, and the biological response to the established biocomposite system was assessed by clinical, histological, histomorphometric, and microtomographic analysis. A comparative analysis revealed that the osteogenic-induced secretome presented an increased diversity of proteins, with emphasis on those related to osteogenesis. Microtomographic and histological morphometric analysis revealed that bioceramic xenografts implanted with secretomes enhanced the new bone formation process, with the osteogenic-induced secretome inducing the highest bone tissue formation. The application of the MSC secretome, particularly from osteogenic-induced populations, may be regarded as an effective therapeutic approach to enhance bone tissue healing and regeneration.

UV photofunctionalization of Zirconia-based materials for abutment fabrication is a promising approach that might influence the formation of a sound peri-implant seal, thus promoting long-term soft and hard tissue implant integration. This study aimed to evaluate the effect of UV treatment of test specimens made by two different ZnO2-based ceramic materials on the hydrophilicity, cell cytotoxicity, and proliferation of human gingival fibroblasts (HGFs). Two Zirconia-based materials, high-translucent and ultra-translucent multi-layered Zirconia (Katana, Kuraray Noritake, Japan), were used to prepare a total of 40 specimens distributed in two equally sized groups based on the material (n = 20). The same surface finishing protocol was applied for all specimens, as suggested by the manufacturer. Half the specimens from each group were treated with UV-C light for 48 h. Water contact angle (WCA), fibroblast cytotoxicity, and proliferation were investigated. The WCA values for the high-translucent Zirconia ranged from 69.9° ± 6.4° to 73.7° ± 13.9° for the treated/non-treated specimens and from 79.5° ± 12.8° to 83.4° ± 11.4° for the ultra-translucent multi-layered Zirconia, respectively. However, the difference was insignificant (F(16) = 3.50, p = 0.292). No significant difference was observed for the fibroblast cytotoxicity test. The results for proliferation revealed a significant difference, which was material-dependent (F(8) = 9.58, p = 0.005). We found that UV surface photofunctionalization of ZrO2-based materials alters the human gingival fibroblast cell viability, which might produce favourable results for cell proliferation.

To facilitate artificial bone construct integration into a patient's body, scaffolds are enriched with different biologically active molecules. Among various scaffold decoration techniques, coating surfaces with cell-derived extracellular matrix (ECM) is a rapidly growing field of research. In this study, for the first time, this technology was applied using primary dental pulp stem cells (DPSCs) and tested for use in artificial bone tissue construction.Rat DPSCs were grown on three-dimensional-printed porous polylactic acid scaffolds for 7 days. After the predetermined time, samples were decellularized, and the remaining ECM detailed proteomic analysis was performed. Further, DPSC-secreated ECM impact to mesenchymal stromal cells (MSC) behaviour as well as its role in osteoregeneration induction were analysed.It was identified that DPSC-specific ECM protein network ornamenting surface-enhanced MSC attachment, migration and proliferation and even promoted spontaneous stem cell osteogenesis. This protein network also demonstrated angiogenic properties and did not stimulate MSCs to secrete molecules associated with scaffold rejection. With regard to bone defects, DPSC-derived ECM recruited endogenous stem cells, initiating the bone self-healing process. Thus, the DPSC-secreted ECM network was able to significantly enhance artificial bone construct integration and induce successful tissue regeneration.DPSC-derived ECM can be a perfect tool for decoration of various biomaterials in the context of bone tissue engineering.

Objectives Millions of people worldwide are affected by diseases or injuries which lead to bone/tooth loss and defects. While such clinical situations are daily practice in most of the hospitals, the widely used treatment methods still have disadvantages. Therefore, this field of medicine is actively searching new tissue regeneration techniques, one of which could be stem cell secretome. Thus, the purpose of this research study was to perform the detail proteomic analysis of periosteum-derived mesenchymal stem cells secretome in order to evaluate if it is capable to induce osteo-regenerative process. Material and Methods Periosteum-derived mesenchymal stem cells (PMSCs) were extracted from adult male New Zealand White rabbits. Cells were characterised by evaluating their differentiation potential. After characterisation PMSCs secretomes were collected and their proteomic analysis was performed. Results PMSCs were extracted from adult male New Zealand White rabbits. In order to characterise the extracted PMSCs, they were differentiated in the directions which mainly describes MSC multipotency - osteogenic, myogenic and adipogenic. A total of 146 proteins were detected. After characterisation PMSCs secretomes were collected and their proteomic analysis was performed. The resulting protein composition indicates the ability to promote bone regeneration to fully mature bone. Conclusions Bioactive molecules detected in periosteum-derived mesenchymal stem cells secretome initiates the processes required for the formation of a fully functional bone.

Background Curcumin, a natural polyphenolic substance, has been known for more than two millennia as having strong anti-inflammatory activity towards multiple ailments, including arthritis. The main drawback of curcumin is its poor solubility in water, which leads to low intestinal absorption and minimal bioavailability. In this study, we aimed to compare the anti-arthritic in vivo effect of different curcumin preparations – basic curcumin extract, micellar curcumin, curcumin mixture with piperine, and microencapsulated curcumin. Methods Arthritis was induced in Wistar rats by complete Freund’s adjuvant, and the severity of arthritis was evaluated daily using the arthritis score system. Curcumin preparations were given to animals per os daily for 20 consecutive days, starting at 6th day after arthritis induction. To determine the inflammatory background, pro-inflammatory cytokines were determined using the ELISA test. In addition, hematologic test, weight change, and limb swelling were tracked. Results Our results indicate that curcumin had a rather weak effect on arthritis progression in the Wistar rat model, microencapsulated curcumin effectively prevented the progression of arthritis – the disease stabilized after 10 days of supplementation. It also reduced the levels of immune cells (neutrophils and leukocytes), as well as pro-inflammatory cytokines – TNFα, IL-1, and IL-6, which levels were close to arthritis-free control. Other formulations of curcumin had lower or no effect on arthritis progression. Conclusion Our study shows that the same concentrations of curcumin had a distinctly expressed positive anti-inflammatory effect depending on the form of its delivery. Specifically, we found that microencapsulated curcumin had the most promising effect for treatment. Graphical abstract

This study aimed to create novel bioceramic coatings on a titanium alloy and evaluate their surface properties in comparison with conventional prosthetic materials. The highly polished titanium alloy Ti6Al4V (Ti) was used as a substrate for yttria-stabilized zirconium oxide (3YSZ) and lithium disilicate (LS2) coatings. They were generated using sol-gel strategies. In comparison, highly polished surfaces of Ti, yttria-stabilized zirconium oxide (ZrO2), polyether ether ketone (PEEK) composite, and poly(methyl methacrylate) (PMMA) were utilized. Novel coatings were characterized by an X-ray diffractometer (XRD) and scanning electron microscope (SEM). The roughness by atomic force microscope (AFM), water contact angle (WCA), and surface free energy (SFE) were determined. Additionally, biocompatibility and human gingival fibroblast (HGF) adhesion processes (using a confocal laser scanning microscope (CLSM)) were observed. The deposition of 3YSZ and LS2 coatings changed the physicochemical properties of the Ti. Both coatings were biocompatible, while Ti-3YSZ demonstrated the most significant cell area of 2630 &mu, m2 (p &le, 0.05) and the significantly highest, 66.75 &plusmn, 4.91, focal adhesions (FAs) per cell after 24 h (p &le, 0.05). By contrast, PEEK and PMMA demonstrated the highest roughness and WCA and the lowest results for cellular response. Thus, Ti-3YSZ and Ti-LS2 surfaces might be promising for biomedical applications.

Topography of the scaffold is one of the most important factors defining the quality of artificial bone. However, the production of precise micro- and nano-structured scaffolds, which is known to enhance osteogenic differentiation, is expensive and time-consuming. Meanwhile, little is known about macro-patterns (larger than cell diameter) effect on cell fate, while this kind of structures would significantly facilitate the manufacturing of artificial skeleton. Therefore, this research is focused on polylactic acid scaffold's macro-pattern impact on rat's dental pulp stem cells (DPSCs) morphology, proliferation, and osteogenic differentiation. For this study, two types of scaffolds were 3D printed: wavy and porous. Wavy scaffolds consisted of 188 μm wide joined threads, meaning that cells might have been curved on the filament as well as compressed in the groove. Porous scaffolds were designed to avoid groove formation and consisted of 500 μm threads, arranged in the woodpile manner, forming 300 μm diameter pores. We found that both macro-surfaces influenced DPSC morphology compared to control. As a consequence, enhanced DPSC proliferation and increased osteogenic differentiation potential was registered in cells grown on these scaffolds. Finally, our results showed that the construction of an artificial bone did not necessarily require the precise structuring of the scaffold, because both types of macro-topographic PLA scaffolds were sufficient enough to induce spontaneous DPSC osteogenic differentiation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 174-186, 2019.

3D printing of polylactic acid (PLA) and hydroxyapatite (HA) or bioglass (BG) bioceramics composites is the most promising technique for artificial bone construction. However, HA and BG have different chemical composition as well as different bone regeneration inducing mechanisms. Thus, it is important to compare differentiation processes induced by 3D printed PLA + HA and PLA + BG scaffolds in order to evaluate the strongest osteoconductive and osteoinductive properties possessing bioceramics. In this study, we analysed porous PLA + HA (10%) and PLA + BG (10%) composites' effect on rat's dental pulp stem cells fate in vitro. Obtained results indicated, that PLA + BG scaffolds lead to weaker cell adhesion and proliferation than PLA + HA. Nevertheless, osteoinductive and other biofriendly properties were more pronounced by PLA + BG composites. Overall, the results showed a strong advantage of bioceramic BG against HA, thus, 3D printed PLA + BG composite scaffolds could be a perspective component for patient-specific, cheaper and faster artificial bone tissue production.

Topography of the scaffold is one of the most important factors defining the quality of artificial bone. However, the production of precise micro- and nano-structured scaffolds, which is known to enhance osteogenic differentiation, is expensive and time-consuming. Meanwhile, little is known about macro-patterns (larger than cell diameter) effect on cell fate, while this kind of structures would significantly facilitate the manufacturing of artificial skeleton. Therefore, this research is focused on polylactic acid scaffold's macro-pattern impact on rat's dental pulp stem cells (DPSCs) morphology, proliferation, and osteogenic differentiation. For this study, two types of scaffolds were 3D printed: wavy and porous. Wavy scaffolds consisted of 188 μm wide joined threads, meaning that cells might have been curved on the filament as well as compressed in the groove. Porous scaffolds were designed to avoid groove formation and consisted of 500 μm threads, arranged in the woodpile manner, forming 300 μm diameter pores. We found that both macro-surfaces influenced DPSC morphology compared to control. As a consequence, enhanced DPSC proliferation and increased osteogenic differentiation potential was registered in cells grown on these scaffolds. Finally, our results showed that the construction of an artificial bone did not necessarily require the precise structuring of the scaffold, because both types of macro-topographic PLA scaffolds were sufficient enough to induce spontaneous DPSC osteogenic differentiation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 174-186, 2019.