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Human cell line HS-27A represents an immortalized subpopulation of human stromal cells derived from bone marrow. HS-27A cells meet the criteria set by the International Society of Cell Therapy (ISCT) for the classification as mesenchymal stem cells (MSCs) by expression of surface molecules CD73, CD90, CD105 and HLA-ABC with no expression of CD14, CD31, CD34, CD45 and HLA-DR. We hypothesized that these cells may undergo osteogenesis similar to human bone marrow-derived stromal cells (BMSCs) and serve as a model of osteogenic cell responses under normal and inflammatory conditions. HS-27A cells were treated with: standard osteogenic factors, i.e., ascorbic acid (Asc), dexamethasone (Dex) and β-glycerophosphate (BGP); recombinant human bone morphogenetic protein 2 (rhBMP-2), MEK1/2 kinase inhibitor (PD98059) and/or lipopolysaccharide (LPS). Cells were examined for alkaline phosphate (ALP) activity, mRNA expression of osteoblastic markers (qRT-PCR) and mineralization of extracellular matrix. Besides, we analyzed mRNA expression of proinflammatory cytokines IL-1β and TNF-α in LPS-treated cells and evaluated transfection efficiency of these cells with Lipofectamine 3000 for potential future genetic manipulations. We determined that HS-27A cell line increase alkaline phosphatase activity (P<0.05), while mineralization of extracellular matrix remains low. Cells treatment with rhBMP-2 and PD98059 resulted in increased mRNA levels for osteogenesis-related transcription factor MSX-2, bone sialoprotein and osteocalcin (all P<0.05). In osteogenic cultures with and without rhBMP-2, addition of LPS led to increased ALP activity, mRNA levels for collagen type I and osteocalcin as well for IL-1β and TNF-α (all P<0.05). We also show that HS-27A cells can be transfected with phMGFP plasmid using Lipofectamine 3000 with low efficiency that may be sufficient for some genetic manipulations. Thus, HS-27A cell line appear as a useful in vitro cell culture model to study short-term osteogenic and inflammatory-related responses of human bone marrow stromal cells.

In this work, we show the in vitro anticancer potential of surgical wires, obtained from zinc (ZnMg0.004) or magnesium (MgCa0.7) alloys by spatial technology comprising casting, extrusion, and final drawing processes. We also present the selective anticancer effects of applied soluble multilayer nanocoatings of zinc and magnesium onto titanium surfaces using the pulse laser deposition method. In the latter, the titanium samples were produced via 3D printing using the selective laser melting method and coated with various combinations of zinc and magnesium layers. For cytotoxicity studies, human dental pulp-derived stem cells (hDPSCs) and human osteosarcoma SaOS-2 cell line were used as representatives of healthy and cancer cells. Cells were examined against the 0.3-3.0 cm 2 /mL material extract ratios obtained from experimental and steel surgical wires, the latter being the current clinical industry standard. The MgCa0.7 alloy wires were approx. 1.5 times more toxic to cancer cells at all examined extract ratios vs. the extracts from steel surgical wires that exhibited comparable toxicity towards healthy and cancer cells. The ZnMg0.004 alloy wires displayed increased toxicity towards cancer cells with decreasing extract ratios. This was also reflected in the increased anticancer effectiveness, calculated based on the viability ratio of healthy cells to cancer cells, from 1.1 to 4.0 times. Healthy cell viability remained at 80-100%, whereas cancer cell survival fluctuated at 20-75%, depending on the extract ratio. Furthermore, the culture of normal or cancer cells on the surface of Zn/Mg-coated titanium allowed us to select combinations of specific coating layers that yielded a comparable anticancer effectiveness to that observed with the experimental wires that ranged between 2 and 3. Overall, this work not only demonstrates the substantial anticancer properties of the studied wires but also indicates that similar anticancer effects can be replicated with appropriate nanocoatings on titanium samples. We believe that this work lays the groundwork for the future potential development of the category of new implants endowed with anticancer properties.

The balance between Noggin and bone morphogenetic proteins (BMPs) is important during early development and skeletal regenerative therapies. Noggin binds BMPs in the extracellular space, thereby preventing BMP signaling. However, Noggin may affect cell response not necessarily through the modulation of BMP signaling, raising the possibility of direct Noggin signaling through yet unspecified receptors. Here we show that in osteogenic cultures of adipose-derived stem cells (ASCs), Noggin activates fibroblast growth factor receptors (FGFRs), Src/Akt and ERK kinases, and it stabilizes TAZ proteins in the presence of dexamethasone. Overall, this leads ASCs to increased expression of osteogenic markers and robust mineral deposition. Our results also indicate that Noggin can induce osteogenic genes expression in normal human bone marrow stem cells and alkaline phosphatase activity in normal human dental pulp stem cells. Besides, Noggin can specifically activate FGFR2 in osteosarcoma cells. We believe our findings open new research avenues to further explore the involvement of Noggin in cell fate modulation by FGFR2/Src/Akt/ERK signaling and potential applications of Noggin in bone regenerative therapies., (© 2024. The Author(s).)

In this work, the biological properties of three-dimensional scaffolds based on a blend of nanohydroxyapatite (nHA), silk fibroin (SF), and chitosan (CTS), were prepared using a lyophilization technique with various weight ratios: 10:45:45, 15:15:70, 15:70:15, 20:40:40, 40:30:30, and 70:15:15 nHA:SF:CTS, respectively. The basic 3D scaffolds were obtained from 5% ( w / w ) chitosan and 5% silk fibroin solutions and then nHA was added. The morphology and physicochemical properties of scaffolds were studied and compared. A biological test was performed to study the growth and osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs). It was found that the addition of chitosan increases the resistance properties and extends the degradation time of materials. In vitro studies with human mesenchymal stem cells found a high degree of biotolerance for the materials produced, especially for the 20:40:40 and 15:70:15 (nHa:SF:CTS) ratios. The presence of silk fibroin and the elongated shape of the pores positively influenced the differentiation of cells into osteogenic cells. By taking advantage of the differentiation/proliferation cues offered by individual components, the composites based on the nanohydroxyapatite, silk fibroin, and chitosan scaffold may be suitable for bone tissue engineering, and possibly offer an alternative to the widespread use of collagen materials.

In this work, a poly(L-lactide- co -glycolide) (PLGA)-based composite was enriched with one of the following sol-gel bioactive glasses (SBG) at 50 wt.%: A1-40 mol% SiO 2 , 60 mol% CaO, CaO/SiO 2 ratio of 1.50; S1-80 mol% SiO 2 , 20 mol% CaO, CaO/SiO 2 ratio of 0.25; A2-40 mol% SiO 2 , 54 mol% CaO, 6 mol% P 2 O 5 , CaO/SiO 2 ratio of 1.35; S2-80 mol% SiO 2 ,16 mol% CaO, 4 mol% P 2 O 5 , CaO/SiO 2 ratio of 0.20. The composites and PLGA control sheets were then soaked for 24 h in culture media, and the obtained condition media (CM) were used to treat human bone marrow stromal cells (hBMSCs) for 72 h. All CMs from the composites increased ERK 1/2 activity vs. the control PLGA CM. However, expressions of cell migration-related c-Fos, osteopontin, matrix metalloproteinase-2, C-X-C chemokine receptor type 4, vascular endothelial growth factor, and bone morphogenetic protein 2 were significantly increased only in cells treated with the CM from the A1/PLGA composite. This CM also significantly increased the rate of human BMSC migration but did not affect cell metabolic activity. These results indicate important biological markers that are upregulated by products released from the bioactive composites of a specific chemical composition, which may eventually prompt osteoprogenitor cells to colonize the bioactive material and accelerate the process of tissue regeneration.

Hydrogels based on collagen/β-glucan crosslinked with tannic acid were obtained by neutralization using dialysis. The presence of tannic acid allowed obtaining stable hydrogel materials with better mechanical properties. Tannic acid was released from matrices gradually and not rapidly. The antioxidant properties of the obtained hydrogels increased over the course of their incubation in culture media and were dependent on the concentration of tannic acid in the matrices. The obtained materials influenced dehydrogenase activity and the ATP level of pathogens. Additionally, the materials' extracts improved the HaCaT cells' viability. Therefore, the obtained hydrogels seem to be promising biocompatible materials which display antimicrobial properties.

In this study, hydrogels based on chitosan cross-linked by glyoxal have been investigated for potential medical applications. Hydrogels were loaded with tannic acid at different concentrations. The thermal stability and the polyphenol-releasing rate were determined. For a preliminary assessment of the clinical usefulness of the hydrogels, they were examined for blood compatibility and in the culture of human dental pulp cells (hDPC). The results showed that after immersion in a polyphenol solution, chitosan/glyoxal hydrogels remain nonhemolytic for erythrocytes, and we also did not observe the cytotoxic effect of hydrogels immersed in tannic acid (TA) solutions with different concentration. Tannic acid was successfully released from hydrogels, and its addition improved material thermal stability. Thus, the current findings open the possibility to consider such hydrogels in clinics.

Collagen possesses unique properties, e.g., biocompatibility, biodegradability, and non-toxicity. However, collagen material degrades too quickly and has low mechanical properties. One of the methods of polymers' modification is mixing them to obtain blends. In this study, the influence of β-glucan for collagen material was analyzed. The interaction between the functional groups of the polymer was analyzed by ATR-FTIR (attenuated total reflection-fourier transform infrared) spectroscopy. The influence of β-glucan on mechanical properties was evaluated. The surface properties of materials were assessed using contact angle measurements and the topography of materials was evaluated by AFM (atomic force microscope). The structure of materials was analyzed according to SEM (scanning electron microscopy) pictures. Moreover, the DPPH-free radicals' scavenging ability and biocompatibility against erythrocytes and HaCaT cells were evaluated. Collagen and β-glucan were bound together by a hydrogen bond. β-glucan addition increased the roughness of the surface of the film and resulted in a more rigid character of the materials. A small addition of β-glucan to collagen provided a more hydrophilic character. All the materials could swell in in vitro conditions and showed antioxidant activity. Materials do not cause erythrocyte hemolysis. Finely, our cytotoxicity studies indicated that β-glucan can be safely added at small (10% or less) quantity to collagen matrix, they sufficiently support cell growth, and the degradation products of such matrices may actually provide some beneficial effects to the surrounding cells/tissues.

The aim of this work was to study effect of the type of silica nanoparticles on the properties of nanocomposites for application in the guided bone regeneration (GBR). Two types of nanometric silica particles with different size, morphology and specific surface area (SSA) i.e., high specific surface silica (hss-SiO 2 ) and low specific surface silica (lss-SiO 2 ), were used as nano-fillers for a resorbable polymer matrix: poly(L-lactide-co-D,L-lactide), called PLDLA. It was shown that higher surface specific area and morphology (including pore size distribution) recorded for hss-SiO 2 influences chemical activity of the nanoparticle; in addition, hydroxyl groups appeared on the surface. The nanoparticle with 10 times lower specific surface area (lss-SiO 2 ) characterized lower chemical action. In addition, a lack of hydroxyl groups on the surface obstructed apatite nucleation (reduced zeta potential in comparison to hss-SiO 2 ), where an apatite layer appeared already after 48 h of incubation in the simulated body fluid (SBF), and no significant changes in crystallinity of PLDLA/lss-SiO 2 nanocomposite material in comparison to neat PLDLA foil were observed. The presence and type of inorganic particles in the PLDLA matrix influenced various physicochemical properties such as the wettability, and the roughness parameter note for PLDLA/lss-SiO 2 increased. The results of biological investigation show that the bioactive nanocomposites with hss-SiO 2 may stimulate osteoblast and fibroblast cells'proliferation and secretion of collagen type I. Additionally, both nanocomposites with the nanometric silica inducted differentiation of mesenchymal cells into osteoblasts at a proliferation stage in in vitro conditions. A higher concentration of alkaline phosphatase (ALP) was observed on the material modified with hss-SiO 2 silica.

In this work, we examined the in vitro cytotoxicity of new biodegradable surgical wires. The wires made of zinc with the addition of a small amount of magnesium (pure zinc, ZnMg 0.0026, ZnMg 0.0068, and ZnMg 0.08) have been investigated. The wires were produced using a technology based on extrusion and subsequent drawing. The resulting wires with a diameter of 0.8-1.0 mm are designed to be used in surgical operations related to bone joints. For cytotoxicity studies, we have selected human dental pulp stem cells (hDPSC) as the cell population representing normal osteoprogenitor cells. Considering that, after bone surgeries, the chance of osteosarcoma increases, we have compared the results obtained in hDPSC to those obtained with Saos-2 human osteosarcoma cell line. Cultured cells were exposed to the extracts obtained from the materials incubated in culture medium for 24 h with and without preincubation. Extracts of different ratios were examined. The results showed that the extracts obtained from wires made of ZnMg 0.0026 alloy exhibit high toxicity to Saos-2 osteosarcoma cells and low toxicity to hDPSC cells. This was in contrast to all reference materials, i.e., commercial surgical sutures made of steel and polymers, that did not display cytotoxicity toward osteosarcoma cells. Thus, the detected phenomenon for the ZnMg 0.0026 alloy can become the basis for creating biodegradable Zn-Mg surgical wires with antitumor activity.

Materials based on carbohydrate polymers may be used for biomedical application. However, materials based on natural polymers have weak physicochemical properties. Thereby, there is a challenge to improve their properties without initiation of toxicity. The alternative method compared to toxic chemical agents' addition is the use of metal complexation method. In this study, chitosan/tannic acid mixtures modified by Fe(III) complexation are proposed and tested for potential applications as wound dressings. Thereby, surface properties, blood compatibility as well as platelet adhesion was tested. In addition, the periodontal ligament stromal cells compatibility studies were carried out. The results showed that the iron(III) addition to chitosan/tannic acid mixture improves properties due to a decrease in the surface free energy and exhibited a reduction in the hemolysis rate (below 5%). Moreover, cells cultured on the surface of films with Fe(III) showed higher metabolic activity. The current findings allow for the medical application of the proposed materials as wound dressings.

Scaffolds based on chitosan (CTS), collagen (Coll) and glycosaminoglycans (GAG) mixtures cross-linked by tannic acid (TA) with bioglass 45S5 addition were obtained with the use of the freeze-drying method. The prepared scaffolds were characterised for morphology, mechanical strength and degradation rate. Moreover, cell viability on the obtained scaffolds was measured with and without the presence of ascorbic acid and dexamethasone. The main purpose of the research was to compare the effectiveness of bioglass 45S5 influence on the physicochemical and biological properties of scaffolds. The results demonstrated that the scaffolds based on the blends of biopolymers cross-linked by TA are stable in an aqueous environment. Scanning electron microscope images allowed the observation of a porous scaffold structure with interconnected pores. The addition of bioglass nanoparticles improved the mechanical properties and decreased the degradation rate of composite materials. The biological properties were improved for 20% tannic acid addition compared to 5%. However, the addition of bioglass 45S5 did not change to cells response significantly.

Sodium alginate and tannic acid are natural compounds that can be mixed with each other. In this study, we propose novel eco-friendly hydrogels for biomedical applications. Thus, we conducted the following assessments including (i) observation of the structure of hydrogels by scanning electron microscope; (ii) bioerosion and the concentration of released tannic acid from subjected material; (iii) dehydrogenase activity assay to determine antibacterial activity of prepared hydrogels; and (iv) blood and cell compatibility. The results showed that hydrogels based on sodium alginate/tannic acid exert a porous structure. The immersion in simulated body fluid (SBF) results in the biomineralization process occurring on their surface while the bioerosion studies revealed that the addition of tannic acid improves hydrogels' stability proportional to its concentration. Besides, tannic acid release concentration depends on the type of hydrogels and the highest amount was noticed for those based on sodium alginate with the content of 30% tannic acid. Antibacterial activity of hydrogels was proven for both Gram-negative and Gram-positive bacteria, the hemolysis rate was below 5% and the viability of the cells was elevated with an increasing amount of tannic acid in hydrogels. Collectively, we assume that obtained materials make the imperative to consider them for biomedical applications.

The interests in the biomedical impact of tannic acid (TA) targeting production of various types of biomaterials, such as digital microfluids, chemical sensors, wound dressings, or bioimplants constantly increase. Despite the significant disadvantage of materials obtained from natural-based compounds and their low stability and fragility, therefore, there is an imperative need to improve materials properties by addition of stabilizing formulas. In this study, we performed assessments of thin films over TA proposed as a cross-linker to be used in combination with polymeric matrix based on chitosan (CTS), i.e. CTS/TA at 80:20 or CTS/TA at 50:50 and poly(ethylene glycol) (PEG) at the concentration of 10% or 20%. We evaluated their mechanical parameters as well as the cytotoxicity assay for human bone marrow mesenchymal stem cells, human melanotic melanoma (MNT-1), and human osteosarcoma (Saos-2). The results revealed significant differences in dose-dependent of PEG regarding the maximum tensile strength (σ max ) or impact on the metabolic activity of tissue culture plastic. We observed that PEG improved mechanical parameters prominently, decreased the hemolysis rate, and did not affect cell viability negatively. Enclosed data, confirmed also by our previous reports, will undoubtedly pave the path for the future application of tannic acid-based biomaterials to treat wound healing.

Spacers, in terms of instruments used in revision surgery for the local treatment of postoperative infection, are usually made of metal rod covered by antibiotic-loaded bone cement. One of the main limitations of this temporary implant is the debonding effect of metal-bone cement interface, leading to aseptic loosening. Material selection, as well as surface treatment, should be evaluated in order to minimize the risk of fraction and improve the implant-cement fixation the appropriate manufacturing. In this study, Ti13Zr13Nb alloys that were prepared by Selective Laser Melting and surface treated were coated with bone cement loaded with either gentamicin or nanosilver, and the effects of such alloy modifications were investigated. The SLM-made specimens of Ti13Zr13Nb were surface treated by sandblasting, etching, or grounding. For each treatment, Scanning Electron Microscope (SEM), contact profilometer, optical tensiometer, and nano-test technique carried out microstructure characterization and surface analysis. The three types of bone cement i.e., pure, containing gentamicin and doped with nanosilver were applied to alloy surfaces and assessed for cement cohesion and its adhesion to the surface by nanoscratch test and pull-off. Next, the inhibition of bacterial growth and cytocompatibility of specimens were investigated by the Bauer-Kirby test and MTS assay respectively. The results of each test were compared to the two control groups, consisting of commercially available Ti13Zr13Nb and untreated SLM-made specimens. The highest adhesion bone cement to the titanium alloy was obtained for specimens with high nanohardness and roughness. However, no explicit relation of adhesion strength with wettability and surface energy of alloy was observed. Sandblasting or etching were the best alloys treatments in terms of the adhesion of either pure or modified bone cements. Antibacterial additives for bone cement affected its properties. Gentamicin and nanosilver allowed for adequate anti-bacterial protection while maintaining the overall biocompatibility of obtained spacers. However, they had different effects on the cement's adhesive capacity or its own cohesion. Furthermore, the addition of silver nanoparticles improved the nanomechanical properties of bone cements. Surface treatment and method of fabrication of titanium affected surface parameters that had a significant impact on cement-titanium fixation.

Bacterial infections due to bone replacement surgeries require modifications of bone cement with antibacterial components. This study aimed to investigate whether the incorporation of gentamicin or nanometals into bone cement may reduce and to what extent bacterial growth without the loss of overall cytocompatibility and adverse effects in vitro. The bone cement Cemex was used as the base material, modified either with gentamicin sulfate or nanometals: Silver or copper. The inhibition of bacterial adhesion and growth was examined against five different bacterial strains along with integrity of erythrocytes, viability of blood platelets, and dental pulp stem cells. Bone cement modified with nanoAg or nanoCu revealed greater bactericidal effects and prevented the biofilm formation better compared to antibiotic-loaded bone cement. The cement containing nanoAg displayed good cytocompatibility without noticeable hemolysis of erythrocytes or blood platelet disfunction and good viability of dental pulp stem cells (DPSC). On the contrary, the nanoCu cement enhanced hemolysis of erythrocytes, reduced the platelets aggregation, and decreased DPSC viability. Based on these studies, we suggest the modification of bone cement with nanoAg may be a good strategy to provide improved implant fixative for bone regeneration purposes.

Polymeric surfaces suitable for cell culture (DR/Pec) were constructed from diazoresin (DR) and pectin (Pec) in a form of ultrathin films using the layer-by-layer (LbL) technique. The surfaces were functionalized with insulin using diazonium chemistry. Such functionalized surfaces were used to culture human mesenchymal stem cells (hMSCs) to assess their suitability for bone tissue engineering and regeneration. The activity of insulin immobilized on the surfaces (DR/Pec/Ins) was compared to that of insulin dissolved in the culture medium. Human MSC grown on insulin-immobilized DR/Pec surfaces displayed increased proliferation and higher osteogenic activity. The latter was determined by means of alkaline phosphatase (ALP) activity, which increases at early stages of osteoblasts differentiation. Insulin dissolved in the culture medium did not stimulate cell proliferation and its osteogenic activity was significantly lower. Addition of recombinant human bone morphogenetic protein 2 (rhBMP-2) to the culture medium further increased ALP activity in hMSCs indicating additive osteogenic action of immobilized insulin and rhBMP-2.

Objectives: Properties of cell culture supports obtained from ultrathin multilayer films containing anionic natural polysaccharides (PSacs) and a synthetic polycation were studied., Materials and Methods: Supports were prepared via a layer-by-layer (LbL) self-assembly deposition method. Polymers used were: heparin (Hep), chondroitin sulphate (CS), hyaluronic acid (HA), and ι-carrageenan (Car) as polyanions, and diazoresin (DR) as a polycation. PSac layers were crosslinked with DR layers by irradiation with UV light absorbed by DR resin., Results: DR/PSac films are very efficient cell culture growth supports as found from experiments with human mesenchymal stem cells (hMSCs). Irradiation of the films resulted in changing zeta potential of outermost layers of both DR and PSac to more negative values, and in increased film hydrophobicity, as found from the contact angle measurements. Photocrosslinking of the supports led to their increased stability., Conclusions: The supports allow for obtaining intact cell monolayers faster than when typical polystyrene tissue culture plates are used. Moreover, these monolayers spontaneously detach permitting formation of new cell layers on these surfaces relatively early during culture, compared to cells cultured on commonly used tissue culture plastic., (© 2014 John Wiley & Sons Ltd.)

Bone marrow stromal cells (MSC) are the major source of osteoblasts for bone remodeling and repair in postnatal animals. Rodent MSC cultured with bone morphogenetic proteins (BMPs) differentiate into osteoblasts, but most human MSC show a poor osteogenic response to BMPs. In this study we demonstrate that BMP-induced osteogenesis in poorly responsive human MSC requires modulation of ERK and phosphatidylinositol 3-kinase (PI3-K) pathways. Either treating human MSC cultures with the MAPK/ERK kinase inhibitor PD98059 or transferring them to serum-free medium with insulin or IGF-I permits BMP-dependent increases in the expression of the early osteoblast-associated genes, alkaline phosphatase and osteopontin. Increased expression of these genes in BMP-treated, serum-free cultures correlates with increased nuclear levels of activated Smads, whereas serum-free cultures of human MSC expressing constitutively active MAPK/ERK kinase show decreased expression of early osteoblast genes and decreased nuclear translocation of BMP-activated Smads. Inhibiting ERK activity in human MSC also elevates the expression of Msx2, a transcription factor that is directly regulated by Smad-binding elements in its promoter. Therefore, growth factor stimulation leading to high levels of ERK activity in human MSC results in suppressed BMP-induced transcription of several early osteoblast genes, probably because levels of BMP-activated nuclear Smads are decreased. In contrast, inhibiting the insulin/IGF-I-activated PI3-K/AKT pathway decreases BMP-induced alkaline phosphatase and osteopontin expression in serum-free cultures of human MSC, but increases BMP activation of Smads; thus, PI3-K signaling is required for BMP-induced expression of early osteoblast genes in human MSC either downstream or independent of the BMP-activated Smad signaling pathway.