The risk of bacterial infections is a significant challenge faced frequently in the use of implants or scaffolds for bone treatment. Therefore, this study focusses on the development and characterisation of novel β tricalcium phosphate (βTCP) scaffolds co-doped with silver and silicon, along with composites coated with antibiotic-loaded poly(3-hydroxybutyrate) (P(3HB)) layers. The successful incorporation of silver and silicon dopants while maintaining the formation of βTCP phase was confirmed using X-ray fluorescence (XRF), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), Raman spectroscopy, and scanning electron microscopy/energy-dispersive X-ray spectroscopy (SEM/EDS) analysis. The developed materials demonstrated comparable total and open porosity (∼ 64-70 vol%), suggesting high interconnectivity between pores conducive to nutrient transport and tissue repair. The increase in compressive strength was achieved for both doped (4.73 ± 0.79 MPa) and P(3HB) coated (5.79 ± 0.92 MPa) scaffolds due to the fine bioceramic microstructure and polymeric coating. Silver and silicon-modified βTCP demonstrated enhanced growth inhibition of Gram-negative (Pseudomonas aeruginosa, Escherichia coli) and Gram-positive (Staphylococcus aureus) bacterial strains in vitro compared to the pure βTCP. P(3HB) coatings, enriched with gentamicin or ceftazidime, exhibited burst and sustained release of the antibiotics from the scaffolds up to 120 h further intensifying the bacteria-killing capability, with evident inhibition zones observed in vitro. Moreover, the composites exhibited apatite-forming ability, suggesting their bioactive potential. In vivo evaluation using Caenorhabditis elegans demonstrated the lack of toxicity of the tested materials. The simultaneous incorporation of the dopants and antibiotic-loaded P(3HB) coatings not only offer a dual antibacterial approach but may also facilitate bone regeneration. However, further in vitro and in vivo investigations are needed to assess their potential in clinical application.

Polyhydroxyalkanoates (PHAs) are biocompatible and biodegradable bacterial-origin polyesters, which have recently emerged as a potential prosperous coating for bioceramic scaffolds. However, biopolymers often have properties inferior to commercially available polymers, and thus blending seems to be an efficient method to tailor their characteristics. In our study poly(3-hydroxybutyrate) (P(3HB)) and medium chain length PHA (mcl-PHA) blends were used as coatings on beta tricalcium phosphate (beta TCP) scaffolds. The influence of the coating type on the physicochemical properties of composites was investigated using various techniques such as X-ray diffraction (XRD), attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), thermogravimetric analysis with differential scanning calorimetry (TG/DSC), scanning electron microscopy (SEM), atomic force microscopy (AFM), wettability measurements, ultra high-performance liquid chromatography with and mass spectrometry (UHPLC-MS) and in vitro studies. It has been demonstrated that blends with various amounts of brittle P(3HB) and viscous mcl-PHA can serve as coatings on beta TCP scaffolds, modulating their physicochemical properties. Macroporous scaffolds covered with blends possessed high open porosity (similar to 65 vol%) and improved compressive strength (up to 4.9 +/- 0.9 MPa). It has been shown that their wettability can be tailored by modifying composition of blends, making the surface more hydrophobic with increasing amount of mcl-PHA. The highest amount of mcl-PHA was shown to significantly influence the degradation of the composites which may be a valuable feature in the case of customized scaffolds for the controlled release of bioactive substances. Moreover, UHPLC-MS analysis revealed that PHAs degrade to hydroxy acids and their oligomers, which may serve as potentially nourishing compounds for surrounding tissues. According to in vitro biocompatibility tests on mouse preosteoblasts, all evaluated scaffolds were nontoxic. The greater the surface hydrophilicity, the better cell adhesion and proliferation were observed.

The search for new materials for bone regenerative purposes is still ongoing. Therefore, we present a series of newly constructed composites based on β tricalcium phosphate (βTCP) and poly(3-hydroxybutyrate) bacteria-derived biopolymer (P(3HB)) in the form of 3D scaffolds with different pore sizes. To improve the polymer attachment to the βTCP surface, the etching of ceramic sinters, using citric acid, was applied. As expected, pre-treatment led to the increase in surface roughness and the creation of micropores facilitating polymer adhesion. In this way, the durability and compressive strength of the ceramic–polymer scaffolds were enhanced. It was confirmed that P(3HB) degrades to 3-hydroxybutyric acid, which broadens applications of developed materials in bone tissue engineering as this compound can potentially nourish surrounding tissues and reduce osteoporosis. Moreover, to the best of our knowledge, it is one of the first studies where the impact of βTCP/P(3HB) scaffolds on mesenchymal stem cells (MSCs), cultured in lowered (5%) oxygen concentration, was assessed. It was decided to use a 5% oxygen concentration in the culture to mimic the conditions that would be found in damaged bone in a living organism during regeneration. Scaffolds enabled cell migration and sufficient flow of the culture medium, ensuring high cell viability. Furthermore, in composites with etched βTCP, the MSCs adhesion was facilitated by hydrophilic ceramic protrusions which reduced hydrophobicity. The developed materials are potential candidates for bone tissue regeneration. Nevertheless, to confirm this hypothesis, in vivo studies should be performed.

Implantations in orthopedics are associated with a high risk of bacterial infections in the surgery area. Therefore, biomaterials containing antibacterial agents, such as antibiotics, bactericidal ions or nanoparticles have been intensively investigated. In this work, silver decorated β tricalcium phosphate (βTCP)-based porous scaffolds were obtained and coated with a biopolymer—poly(3-hydroxybutyrate)-P(3HB). To the best of our knowledge, studies using silver-doped βTCP and P(3HB), as a component in ceramic-polymer scaffolds for bone tissue regeneration, have not yet been reported. Obtained materials were investigated by high-temperature X-ray diffraction, X-ray fluorescence, scanning electron microscopy with energy dispersive spectroscopy, hydrostatic weighing, compression tests and ultrahigh-pressure liquid chromatography with mass spectrometry (UHPLC-MS) measurements. The influence of sintering temperature (1150, 1200 °C) on the scaffolds’ physicochemical properties (phase and chemical composition, microstructure, porosity, compressive strength) was evaluated. Materials covered with P(3HB) possessed higher compressive strength (3.8 ± 0.6 MPa) and surgical maneuverability, sufficient to withstand the implantation procedures. Furthermore, during the hydrolytic degradation of the composite material not only pure (R)-3-hydroxybutyric acid but also its oligomers were released which may nourish surrounding tissues. Thus, obtained scaffolds were found to be promising bone substitutes for use in non-load bearing applications

Novel bone substitutes such as highly porous ceramic scaffolds can serve as platforms for delivering active molecules. A common problem is to control the release of the drug, therefore, it is beneficial to use a drug-functionalized polymer coating. In this study, beta-tricalcium phosphate-based porous scaffolds were obtained and coated with diclofenac-functionalized biopolymer - poly(3-hydroxyoctanoate) - P(3HO). To the best of our knowledge, studies using P(3HO) as a component in ceramic-polymer based drug delivery system for bone tissue regeneration have not yet been reported. Presented materials were comprehensively investigated by various techniques such as powder X-ray diffraction, scanning electron microscopy with energy dispersive spectroscopy, hydrostatic weighing and compression tests, pH and ionic conductivity measurements, high-performance liquid chromatography and in vitro cytotoxicity studies. The obtained diclofenac-loaded composite was not only characterised by controlled and sustained drug release, but also possessed improved mechanical properties. Moreover, the precipitation of apatite-like forms on its surface was observed after incubation in simulated body fluid, which indicates its bioactive potential. After 24 hours no cytotoxic effect on MC3T3-E1 mouse preosteoblastic cells was confirmed using indirect cytotoxicity studies. Thus, this promising multifunctional composite scaffold can be a promising candidate as an anti-inflammatory drug-delivery system in bone tissue engineering.

Nowadays, regenerative medicine faces a major challenge in providing new, functional materials that will meet the characteristics desired to replenish and grow new tissue. Therefore, this study presents new ceramic-polymer composites in which the matrix consists of tricalcium phosphates covered with blends containing a chemically bounded diclofenac with the biocompatible polymer—poly(3-hydroxyoctanoate), P(3HO). Modification of P(3HO) oligomers was confirmed by NMR, IR and XPS. Moreover, obtained oligomers and their blends were subjected to an in-depth characterisation using GPC, TGA, DSC and AFM. Furthermore, we demonstrate that the hydrophobicity and surface free energy values of blends decreased with the amount of diclofenac modified oligomers. Subsequently, the designed composites were used as a substrate for growth of the pre-osteoblast cell line (MC3T3-E1). An in vitro biocompatibility study showed that the composite with the lowest concentration of the proposed drug is within the range assumed to be non-toxic (viability above 70%). Cell proliferation was visualised using the SEM method, whereas the observation of cell penetration into the scaffold was carried out by confocal microscopy. Thus, it can be an ideal new functional bone tissue substitute, allowing not only the regeneration and restoration of the defect but also inhibiting the development of chronic inflammation.

Development of new composite materials for bone tissue engineering is a constantly growing field of medicine. Therefore there is a continuous need in creating novel materials that can not only regenerate the defected tissue but also nourish it while the healing process progresses. Here we present a novel type of composite material that fulfils these requirements. The study describes creation of a composite with macroporous bioceramic core that is infiltrated with a thin biopolymer layer. The ceramic component, namely tricalcium phosphate (TCP), due to its mechanistic and bioactive properties may promote new bone creation as shown through the in vitro studies. To the best of our knowledge the coating layer was created for the first time from a representative of bacterially derived medium chain length polyhydroxyalkanoate polymers (mcl-PHA), namely polyhydroxyoctanoate (PHO). This polymer layer not only profoundly changed the stress-strain characteristics of the bioceramic foam but also released (R)-3-hydroxyacids and their dimers/trimers to the investigated environment. In the manuscript we have in depth characterised these materials employing a set of basic procedures, through 3D structure reconstruction and finishing with prolonged in vitro experiments.