The Specifics of Neovascularization of Wound Defects in the Oral Mucosa during Its Regeneration under a Piezoelectric Polymer Membrane.

We studied restoration of microvessels in the oral mucosa wound defects under a polymer piezoelectric membrane (group 2) and without it (group 1). The control group included animals with intact mucosa. On day 3, the expression of the vascular endothelial growth factor (VEGF) increased in all experimental groups, while the expression of CD34 increased only in group 2, which attested to intensive neoangiogenesis. On day 7, we observed a decrease in VEGF expression and an increase in CD34 expression that was more pronounced in group 2, which reflected the beginning of blood vessels maturation. More rapid formation and maturation of blood vessels in group 2 was confirmed by electron microscopy: on day 7, endothelial cells with mature organelles and signs of active transcapillary exchange were seen. On day 12, the immature blood vessels still predominated in group 1, while in group 2, the expression of angiogenesis markers decreased though remained above the control, which created prerequisites for the complete restoration of wound area vascularization in group 2. In group 1, the expression of VEGF and CD34 was significantly below the control, which attested to the development of poorly vascularized scar tissue.

Hydrolysis of Bone-Replacing Materials Based on Polylactic Acid and Containing Hydroxyapatite in an In Vitro Experiment.

We studied the features of hydrolytic degradation of polylactic acid (PLLA) implants depending on their structural filling with hydroxyapatite (HA). The resistance to in vitro hydrolysis was tested for the following samples: PLLA without HA (control; group 1), PLLA/HA 25 wt% (group 2), and PLLA/HA 50 wt% (group 3). Samples were incubated at 37°C. In the hydrolysate, lactate, calcium ions, and inorganic phosphate were determined. Additionally, the time of appearance of visual deformation and sample disintegration was recorded. PLLA degradation was higher in samples saturated with HA. The highest resistance to deformation was noted for samples without HA. Samples with a PLLA/HA 50 wt% demonstrated the maximum degradation of PLLA in combination with lower resistance to deformation and the highest bioavailability of calcium and phosphate. Group 2 samples are most promising for clinical use.

Peculiarities of Cell Seeding on Polylactic Acid-Based Scaffolds Fabricated Using Electrospinning and Solution Blow Spinning Technologies.

We studied the possibility of seeding bone marrow-derived stromal cells onto polylactic acid-based scaffolds fabricated by electrospinning and solution blow spinning technologies. The cells were applied to the scaffolds by dynamic seeding and scaffolds were then cultured in Petri dishes in culture medium for 3 days. Cell migration to the Petri dish surface was noted only for scaffolds fabricated by electrospinning technology, but DAPI staining confirmed the presence of cells in both scaffolds. The mean number of cells in scaffolds fabricated by electrospinning and solution blow spinning was 56±9 and 81±6, respectively. The scaffold fabricated by solution blow spinning was more effectively (p<0.05) colonized by cells due to its more optimal spatial structure.

Biocompatibility and Osseointegration of Calcium Phosphate-Coated and Non-Coated Titanium Implants with Various Porosities.

The aim of the investigation was to study the influence of pore size and the presence of a biologically active calcium phosphate coating in porous 3D printed titanium implants on the process of integration with the bone tissue. Materials and Methods: Samples of cylindrical implants with three different pore diameters (100, 200, and 400 µm) were fabricated from titanium powder on the Arcam 3D printer (Sweden) using electron beam melting technology. A calcium phosphate coating with a thickness of 20±4 µm was applied to some of the products by microarc oxidation. Cytotoxicity of the implants was determined in vitro on human dermal fibroblast cultures. The samples were implanted in the femoral bones of 36 rabbits in vivo. The animals were divided into 6 groups according to the bone implant samples. The prepared samples and peri-implant tissues were studied on days 90 and 180 after implantation using scanning electron microscopy and histological methods. Results: All samples under study were found to be non-toxic and well biocompatible with the bone tissue. There were revealed no differences between coated and non-coated implants of 100 and 200 µm pore diameters in terms of their histological structure, intensity of vascularization in the early stages, and bone formation in the later stages. Samples with pore diameters of 100 and 200 µm were easily removed from the bone tissue, the depth of bone growth into the pores of the implant was lower than in the samples with pore diameter of 400 µm (p<0.001). There were differences between coated and non-coated samples of 400 µm pore diameter, which was expressed in a more intensive osseointegration of samples with calcium phosphate coating (p<0.05). Conclusion: The optimal surface characteristics of the material for repairing bone defects are a pore diameter of 400 µm and the presence of a calcium phosphate coating.

Solution blow spinning of PLLA/hydroxyapatite composite scaffolds for bone tissue engineering.

Composite poly-L-lactide acid-based scaffolds with hydroxyapatite (HAp) content up to 75 wt.% were fabricated via solution blow spinning. The influence of HAp concentration on structure, wettability, mechanical properties and chemical and phase composition of the produced materials was examined. It was found that with an increase of HAp content the average fiber diameter was increased, the uniaxial strength and relative elongation were reduced, while the phase composition and surface wettability did not change. The performance of the scaffolds during implantation in the parietal bone of a rat skull for a period from 15 to 90 days was studied. The materials have shown high ability to integrate with both soft and hard tissues. It was found that scaffolds with 25 wt.% HAp content significantly enhance osteogenesis during scarification (damage) of the periosteum. Overall, the fabricated scaffolds proved to be highly efficient for replacing bone defects in long tubular bones.

"Solvent/non-solvent" treatment as a method for non-covalent immobilization of gelatin on the surface of poly(l-lactic acid) electrospun scaffolds.

In the present study, we report a simple and efficient method of gelatin immobilization on the surface of PLA electrospun fibers using pre-treatment with a mixture of toluene and ethanol allowing to form swelled surface layer followed by gelatin adsorption from its solution in PBS. Our results demonstrate that gelatin immobilization leads to a decrease in the water contact angle from 120° to 0°, enhances scaffold strength up to 50%, and doubles the number of adhered cells and their average area. We observed that the maximum amount of gelatin (0.07 ± 0.01 mg per cm3 of the scaffold) was immobilized during the first five minutes of exposure to the gelatin solution. Modified scaffolds demonstrated increased strength.

Flexible intramedullary nails for limb lengthening: a comprehensive comparative study of three nails types.

This study aims to investigate the comparative study of calcium phosphate coatings for flexible intramedullary nails (FINs) used to lengthen long tubular bones. The presence of a calcium phosphate coating deposited by micro-arc oxidation (MAO) or a composite coating based on a co-polymer of vinylidene fluoride with tetrafluoroethylene (VDF-TeFE) and hydroxyapatite (HA) on the surface of the FIN significantly enhanced the regeneration of bone in the area of osteotomy during limb lengthening by combined osteosynthesis. The investigation of the physico-chemical properties of the FIN coated with calcium phosphate via MAO demonstrated that the improved bone tissue formation resulted from favourable conditions for adhesion, proliferation and differentiation of multipotent stem cells into osteoblasts on the coating surface. The composite coatings only stimulated the formation of bone tissue in vivo, primarily because of the piezoelectric properties of the VDF-TeFE co-polymer.

Surface modification of electrospun poly-(l-lactic) acid scaffolds by reactive magnetron sputtering.

In this study, we modified the surface of bioresorbable electrospun poly-(l-lactic) acid (PLLA) scaffolds by reactive magnetron sputtering of a titanium target under a nitrogen atmosphere. We examined the influence of the plasma treatment time on the structure and properties of electrospun PLLA scaffolds using SEM, XRF, FTIR, XRD, optical goniometry, and mechanical testing. It was observed that the coating formed did not change physicomechanical properties of electrospun PLLA scaffolds and simultaneously, increased their hydrophilicity. No adverse tissue reaction up to 3 months after subcutaneous implantation of the modified scaffolds was detected in in-vivo rat model. The rate of scaffold replacement by the recipient tissue in-vivo was observed to depend on the plasma treatment time.

Osteoinductive composite coatings for flexible intramedullary nails.

This work presents composite coatings based on a copolymer of vinylidene fluoride with tetrafluoroethylene (VDF-TeFE) and hydroxyapatite (HA) for flexible intramedullary nails (FIN). The effect of the proportion of VDF-TeFE (100-25% wt.) on physicochemical and biological properties of the composite coatings was investigated. It was shown that a decrease of VDF-TeFE in the coating hinders its crystallization in ß and γ forms which have piezoelectric properties. The decrease also reduces an adhesive strength to 9.9±2.4MPa and a relative elongation to 5.9±1.2%, but results in increased osteogenesis. It was demonstrated that the composite coatings with 35% VDF-TeFE has the required combination of physicochemical properties and osteogenic activity. Comparative studies of composite coatings (35% VDF-TeFE) and calcium phosphate coatings produced using micro-arc oxidation, demonstrated comparable results for strength of bonding of these FINs with trabecular bones (~530MPa). It was hypothesized that the high osteoinductive properties of the composite coatings are due to their piezoelectric properties.

[Effect of radio frequency discharge plasma on surface properties and biocompatibility of polycaprolactone matrices].

Surface modification of bioresorbable polymer material (polycaprolactone, PCL) with abnormal glow discharge, initiated during radio-frequency magnetron sputtering of a hydroxyapatite target was investigated. Plasma treatment resulted in an increase of surface roughness of PCL, crystallite size, the surface free energy and hydrophilicity. Increased treatment time (30, 60, 150 seconds) provoked the polymer surface saturation with the sputtering target ions (calcium, phosphorus). The assessment of plasma exposure of PCL surface on bone marrow multipotent mesenchymal stromal cells behavior (BM MSCs) has been performed. Modification of the polymer surface with the abnormal glow discharge stimulated adhesion and subsequent proliferation of BM MSCs; thus, maximum values were achieved with the surface treatment for 60 s. This type of plasma modification did not affect cell viability (apoptosis, necrosis). Thus, the surface modification with abnormal glow discharge, initiated during radio-frequency magnetron sputtering of a hydroxyapatite target, appear to be a promising method of surface modification of bioresorbable polymer material (PCL) for tissue engineering.

Ferroelectric polymer scaffolds based on a copolymer of tetrafluoroethylene with vinylidene fluoride: fabrication and properties.

A solution blow spinning technique is a method developed recently for making nonwoven webs of micro- and nanofibres. The principal advantage of this method compared to a more traditional electrospinning process is its significantly higher production rate. In this work, the solution blow spinning method was further developed to produce nonwoven polymeric scaffolds based on a copolymer of tetrafluoroethylene with vinylidene fluoride solution in acetone. A crucial feature of the proposed method is that high-voltage equipment is not required, which further improves the method's economics. Scanning electron microscopy analysis of the samples demonstrated that the surface morphology of the nonwoven materials is dependent on the polymer concentration in the spinning solution. It was concluded that an optimum morphology of the nonwoven scaffolds for medical applications is achieved by using a 5% solution of the copolymer. It was established that the scaffolds produced from the 5% solution have a fractal structure and anisotropic mechanical properties. X-ray diffraction, infrared spectroscopy, Raman spectroscopy and differential scanning calorimetry demonstrated that the fabricated nonwoven materials have crystal structures that exhibit ferroelectric properties. Gas chromatography has shown that the amount of acetone in the nonwoven material does not exceed the maximum allowable concentration of 0.5%. In vitro analysis, using the culture of motile cells, confirmed that the nonwoven material is non-toxic and does not alter the morpho-functional status of stem cells for short-term cultivation, and therefore can potentially be used in medical applications.