J. Funct. Biomater., Volume 12, Issue 1 (March 2021) – 20 articles

In addition to biocompatibility, an ideal scaffold for the regeneration of valvular tissue should also replicate the natural heart valve extracellular matrix (ECM) in terms of biomechanical properties and structural stability. In our previous paper, we demonstrated the development of collagen type I and hyaluronic acid (HA)-based scaffolds with interlaced microstructure. Such hybrid scaffolds were found to be compatible with cardiosphere-derived cells (CDCs) to potentially regenerate the diseased aortic heart valve. This paper focused on the quantification of the effect of crosslinking density on the mechanical properties under dry and wet conditions as well as degradation resistance. Elastic moduli increased with increasing crosslinking densities, in the dry and wet state, for parent networks, whereas those of interlaced scaffolds were higher than either network alone. Compressive and storage moduli ranged from 35 ± 5 to 95 ± 5 kPa and 16 ± 2 kPa to 113 ± 6 kPa, respectively, in the dry state. Storage moduli, in the dry state, matched and exceeded those of human aortic valve leaflets (HAVL). Similarly, degradation resistance increased with increasing the crosslinking densities for collagen-only and HA-only scaffolds. Interlaced scaffolds showed partial degradation in the presence of either collagenase or hyaluronidase as compared to when exposed to both enzymes together. These results agree with our previous findings that interlaced scaffolds were composed of independent collagen and HA networks without crosslinking between them. Thus, collagen/HA interlaced scaffolds have the potential to fill in the niche for designing an ideal tissue engineered heart valve (TEHV). Full article

Unique biosynthetic abilities revealed by plants determine in vitro cultures of hairy roots as a suitable source of pharmaceutically relevant bioactive compounds. The basic aim of the study was to examine the applicability of aerogel composed of methyltrimethoxysilane (MTMS) for immobilization of Rindera graeca hairy roots by identifying quantitative effects of biomass proliferation and naphthoquinones extracellular secretion in the aerogel-supported culture system. R. graeca hairy roots were simultaneously cultured for 28-days, as (i) nonimmobilized biomass (reference system), (ii) biomass immobilized on macroporous polyurethane foam (PUF), (iii) biomass with disintegrated MTMS aerogel, (iv) biomass immobilized on polypropylene (PP) fibers (as control), and (v) biomass immobilized on monolithic PP-reinforced MTMS aerogel. MTMS aerogel exhibited high level of biocompatibility toward R. graeca hairy roots which grew into the structure of monolithic aerogel-based constructs. Monolithic MTMS-based constructs significantly promoted the proliferation of hairy roots, resulting in 55% higher fresh mass than the reference system. The highest level of naphthoquinones productivity, i.e., 653 µg g_DW⁻¹, was noted for PUF-supported culture system. Full article

The application of various materials in biomedical procedures has recently experienced rapid growth. One of the areas is the treatment of many of different types of bone-related diseases and disorders by using biodegradable polymer-ceramic composites. We have developed a material based on cryogel polyvinyl alcohol, mineralized with calcium phosphate. Composites were obtained by cyclic freezing-thawing, the synthesis of calcium phosphates was carried out in situ under the influence of microwave radiation with heating and stirring. The components of the composites were determined using the methods of IR-spectroscopy and scanning electron microscopy and electron probe microanalyzer, as well as their morphology and surface properties. The biological compatibility of the material was investigated in vivo for a Wistar rat. The assessment of the quality of bone formation between the cryogel-based implant and the damaged bone was carried out by computed tomography. An improvement in the consolidation of the bone defect is observed in the bone with the composite in comparison with the control bone. Full article

Due to the extremely high incidence of lesions and diseases in aging population, it is critical to put all efforts into developing a successful implant for osteochondral tissue regeneration. Many of the patients undergoing surgery present osteochondral fissure extending until the subchondral bone (corresponding to a IV grade according to the conventional radiographic classification by Berndt and Harty). Therefore, strategies for functional tissue regeneration should also aim at healing the subchondral bone and joint interface, besides hyaline cartilage. With the ambition of contributing to solving this problem, several research groups have been working intensively on the development of tailored implants that could promote that complex osteochondral regeneration. These implants may be manufactured through a wide variety of processes and use a wide variety of (bio)materials. This review aimed to examine the state of the art regarding the challenges, advantages, and drawbacks of the current strategies for osteochondral regeneration. One of the most promising approaches relies on the principles of additive manufacturing, where technologies are used that allow for the production of complex 3D structures with a high level of control, intended and predefined geometry, size, and interconnected pores, in a reproducible way. However, not all materials are suitable for these processes, and their features should be examined, targeting a successful regeneration. Full article

Skin regeneration requires a three-dimensional (3D) scaffold for cell adhesion, growth and proliferation. A type of the scaffold offering a 3D structure is a nonwoven material produced via a melt-blown technique. Process parameters of this technique can be adapted to improve the cellular response. Polylactic acid (PLA) was used to produce a nonwoven scaffold by a melt-blown technique. The key process parameters, i.e., the head and air temperature, were changed in the range from 180–270 °C to obtain eight different materials (MB1–MB8). The relationships between the process parameters, morphology, porosity, thermal properties and the cellular response were explored in this study. The mean fiber diameters ranged from 3 to 120 µm. The average material roughness values were between 47 and 160 µm, whereas the pore diameters ranged from 5 to 400 µm. The calorimetry thermograms revealed a correlation between the temperature parameters and crystallization. The response of keratinocytes and macrophages exhibited a higher cell viability on thicker fibers. The cell-scaffold interaction was observed via SEM after 7 days. This result proved that the features of melt-blown nonwoven scaffolds depended on the processing parameters, such as head temperature and air temperature. Thanks to examinations, the most suitable scaffolds for skin tissue regeneration were selected. Full article

Introduction: Many studies in stem cell biology have demonstrated that dental pulp stem cells (DPSC) may be highly proliferative and capable of pluripotent differentiation into many different tissue types. Recent advances in stem cell research have outlined methods for directing in vitro or in vivo growth, viability, and proliferation, as well as differentiation of DPSC—although much remains to be discovered. Based upon this information, the primary objective of this study was to understand the functional biomaterials needed to more effectively direct DPSC viability, growth, and proliferation. Methods: Using an approved protocol, previously collected and isolated samples of DPSC from an existing repository were used. Previously established stem cell biomarkers (Sox-2, Oct-4, NANOG) from each isolate were correlated with their proliferation rates or doubling times to categorize them into rapid, intermediate, or slow-dividing multipotent DPSC. Growth factors and other functional dental biomaterials were subsequently tested to evaluate DPSC responses in proliferation, viability, and morphology. Results: Differential responses were observed among DPSC isolates to growth factors, including vascular endothelial growth factor (VEGF) and bone morphogenic protein (BMP-2), and functional biomaterials such as mineralized trioxide aggregates (MTA). The responsiveness of DPSC isolates did not correlate with any single factor but rather with a combination of proliferation rate and biomarker expression. Conclusions: These data strongly suggest that some, but not all, DPSC isolates are capable of a robust and significant in vitro response to differentiation stimuli, although this response is not universal. Although some biomarkers and phenotypes that distinguish and characterize these DPSC isolates may facilitate the ability to predict growth, viability, and differentiation potential, more research is needed to determine the other intrinsic and extrinsic factors that may contribute to and modulate these DPSC responses to these functional biomaterials for biotechnology and bioengineering applications. Full article

Biopolymers are widely accepted natural materials in regenerative medicine, and further development of their bioactivities and discoveries on their composition/function relationships could greatly advance the field. However, a concise insight on commonly investigated biopolymers, their current applications and outlook of their modifications for multibioactivity are scarce. This review bridges this gap for professionals and especially freshmen in the field who are also interested in modification methods not yet in commercial use. A series of polymeric materials in research and development uses are presented as well as challenges that limit their efficacy in tissue regeneration are discussed. Finally, their roles in the regeneration of select tissues including the skin, bone, cartilage, and tendon are highlighted along with modifiable biopolymer moieties for different bioactivities. Full article

The bone remodeling field has shifted focus towards the delineation of products with two main critical attributes: internal architectures capable to promote fast cell colonization and good mechanical performance. In this paper, Luffa-fibers and graphene nanoplatelets were proposed as porogen template and mechanical reinforcing agent, respectively, in view of framing 3D products by a one-stage polymer-free process. The ceramic matrix was prepared through a reproducible technology, developed for the conversion of marble resources into calcium phosphates (CaP) powders. After the graphene incorporation (by mechanical and ultrasonication mixing) into the CaP matrix, and Luffa-fibers addition, the samples were evaluated in both as-admixed and thermally-treated form (compact/porous products) by complementary structural, morphological, and compositional techniques. The results confirmed the benefits of the two agents’ addition upon the compact products’ micro-porosity and the global mechanical features, inferred by compressive strength and elastic modulus determinations. For the porous products, overall optimal results were obtained at a graphene amount of <1 wt.%. Further, no influence of graphene on fibers’ ability to generate at high temperatures internal interconnected-channels-arrays was depicted. Moreover, its incorporation led to a general preservation of structural composition and stability for both the as-admixed and thermally-treated products. The developed CaP-reinforced structures sustain the premises for prospective non- and load-bearing biomedical applications. Full article

Calcium phosphate coatings are able to improve the osseointegration process due to their chemical composition, which is similar to that of bone tissues. In this work, to increase the long-term corrosion resistance and to improve the osseointegration process of commercially pure titanium Grade 4 (CpTi G4), biomimetic amorphous calcium phosphate (ACP) coatings were electrodeposited for the first time from an acetate bath with a pH level of 7.0 and a Ca:P ratio of 1.67. ACP coatings were obtained on CpTi G4 substrate subjected to sandblasting and autoclaving using electrochemically assisted deposition at a potential of −3 V relative to the open circuit potential for 30 min at room temperature. SEM, EDS, 2D roughness profiles, amplitude-sensitive eddy current method, and Kelvin scanning probe were used for the surface characterization of the biomaterial under study. In vitro corrosion resistance tests were conducted for 21 days in artificial saliva using open circuit potential, polarization curves, and electrochemical impedance spectroscopy measurements. The passive-transpassive behavior was revealed for the obtained ACP coatings. The long-term corrosion resistance test showed a deterioration of the protective properties for CpTi G4 uncoated and coated with ACP with immersion time. The mechanism and kinetics of the pitting corrosion on the CpTi G4|TiO₂|ACP coating system are discussed in detail. Full article

Electrospun ultrathin fibrous scaffold filed with synthetic nanohydroxyapatite (nHAp) and graphene nanoribbons (GNR) has bioactive and osteoconductive properties and is a plausible strategy to improve bone regeneration. Poly(butylene-adipate-co-terephthalate) (PBAT) has been studied as fibrous scaffolds due to its low crystallinity, faster biodegradability, and good mechanical properties; however, its potential for in vivo applications remains underexplored. We proposed the application of electrospun PBAT with high contents of incorporated nHAp and nHAp/GNR nanoparticles as bone grafts. Ultrathin PBAT, PBAT/nHAp, and PBAT/nHAp/GNR fibers were produced using an electrospinning apparatus. The produced fibers were characterized morphologically and structurally using scanning electron (SEM) and high-resolution transmission electron (TEM) microscopies, respectively. Mechanical properties were analyzed using a texturometer. All scaffolds were implanted into critical tibia defects in rats and analyzed after two weeks using radiography, microcomputed tomography, histological, histomorphometric, and biomechanical analyses. The results showed through SEM and high-resolution TEM characterized the average diameters of the fibers (ranged from 0.208 µm ± 0.035 to 0.388 µm ± 0.087) and nHAp (crystallite around 0.28, 0.34, and 0.69 nm) and nHAp/GNR (200–300 nm) nanoparticles distribution into PBAT matrices. Ultrathin fibers were obtained, and the incorporated nHAp and nHAp/GNR nanoparticles were well distributed into PBAT matrices. The addition of nHAp and nHAp/GNR nanoparticles improved the elastic modulus of the ultrathin fibers compared to neat PBAT. High loads of nHAp/GNR (PBATnH5G group) improved the in vivo lamellar bone formation promoting greater radiographic density, trabecular number and stiffness in the defect area 2 weeks after implantation than control and PBAT groups. Full article

The aim of this retrospective study was to clinically evaluate the five-year outcomes of implants placed following a combined approach to the sinus, consisting of sequential drills and osteotomes. Medical records of patients with implants placed in combination with crestal sinus lift using sequential drills and osteotomes, with a residual alveolar bone crest between 4 to 8 mm, and a follow-up of at least five years after final loading, were evaluated. Outcomes were implant and prosthetic survival and success rates, any complication, and marginal bone loss. Data from 96 patients (53 women and 43 men; mean age 54.7 years; range 23–79 years) were collected. A total of 105 single implants were analyzed. After five years of function, two implants were lost and two prostheses failed. No major biological or prosthetic complications occurred. At the five-year examination, the marginal bone loss was 1.24 ± 0.28 mm. Within the limitations of this retrospective study it can be concluded that implants placed following a combined approach to the sinus consisting of sequential drills and osteotomes seem to be a viable option for the treatment of posterior atrophic edentulous maxilla. Full article

Hospital patients and personnel are at risk of nosocomial viral infections, as clearly manifested during the COVID-19 pandemic. Transmission of respiratory viral pathogens can occur through contaminated surfaces, including from medical textiles. Copper has potent biocidal properties, and cuprous oxide impregnated medical textiles (CMT) reduce hospital-acquired bacterial infections. In the current study we confirm the antimicrobial properties of CMT and determine their capacity to reduce infectious titres of human coronavirus (HCoV-229E) in an independent laboratory. The antibacterial and antiviral activities of the CMT were determined according to AATCC TM100-2019 and ISO 18184:2019 standards, respectively. The CMT reduced by 4 logs the viable titers of MRSA, Klebsiella pneumoniae, Enterococcus faecalis, and Candida auris after 2 h of incubation. Viable titers of Clostridium difficile were reduced by 2.3, 3, and 4 logs after 2, 6, and 18 h, respectively. Infectious titers of HCoV-229E exposed to CMT for 2 h were reduced by 2.8 and 4 logs (99.85% and 99.99% reductions) as compared to Time-0 control and initial inoculum, respectively. The CMT retain their antibacterial efficacy even after 100 industrial washings. Use of cuprous oxide impregnated textiles in clinical settings may reduce not only hospital acquired infections caused by bacterial and fungal pathogens, but also, and equally important, those caused by coronavirus and other viruses. Full article

While periprosthetic joint infections (PJIs) result in a small percentage of patients following arthroplasties, they are challenging to treat if they spread into bone and soft tissue. Treatment involves delivering antibiotics using poly(methyl methacrylate) (PMMA) bone cement. However, antibiotic release is insufficient for prolonged infections. Previous work demonstrated efficacy of incorporating insoluble cyclodextrin (CD) microparticles into PMMA to improve antibiotic release and allow for post-implantation drug refilling to occur in a tissue-mimicking model. To simulate how antibiotic refilling may be possible in more physiologically relevant models, this work investigated development of bone and muscle refilling models. The bone refilling model involved embedding PMMA-CD into rabbit femur and administering antibiotic via intraosseous infusion. Muscle tissue refilling model involved implanting PMMA-CD beads in bovine muscle tissue and administering antibiotic via tissue injection. Duration of antimicrobial activity of refilled PMMA-CD was evaluated. PMMA-CD composite in bone and muscle tissue models was capable of being refilled with antibiotics and resulted in prolonged antimicrobial activity. PMMA-CD provided sustained and on-demand antimicrobial activity without removal of implant if infection develops. Intraosseous infusion appeared to be a viable technique to enable refilling of PMMA-CD after implantation in bone, reporting for the first time the ability to refill PMMA in bone. Full article

The aim of this study was to quantitatively evaluate alloplastic Temporomandibular Joint (TMJ) Prostheses against other treatment modalities regarding the jaw kinematics. Six patients with Temporomandibular Joint Prostheses, four with mandibular ramus Patient-Specific Implant (PSI) with condylar head preservation, and four after mandibular condylectomy were evaluated by the means of axiography (Cadiax Compact 2), which is the noninvasive three-dimensional study of condylar movements. The patients were also evaluated clinically for the mandibular movements. The study revealed that the significant movement limitations occurred bilaterally in patients fitted with TMJ prosthesis. For the protrusion movement, the vector length of the movement (L) for the TMJ prosthesis was 0.31 vs. 3.01 mm for the PSI (Kruskal–Wallis chi-squared = 9.1667, df = 2, p-value = 0.01022, post hoc Dunn p-value = 0.015) and for the laterotrusion to the operated side, the length of the vector (L) was 0.66 vs. 3.35 mm, respectively. Statistically significant differences between groups were most frequent for the laterotrusion to the unoperated side. The study shows that a further development on TMJ Prostheses geometry and materials is needed. Full article

Acrylic bone cements modified with linoleic acid are a promising low-modulus alternative to traditional high-modulus bone cements. However, several key properties remain unexplored, including the effect of autoclave sterilization and the potential use of low-modulus cements in other applications than vertebral augmentation. In this work, we evaluate the effect of sterilization on the structure and stability of linoleic acid, as well as in the handling properties, glass transition temperature, mechanical properties, and screw augmentation potential of low-modulus cement containing the fatty acid. Neither ¹H NMR nor SFC-MS/MS analysis showed any detectable differences in autoclaved linoleic acid compared to fresh one. The peak polymerization temperature of the low-modulus cement was much lower (28–30 °C) than that of the high-modulus cement (67 °C), whereas the setting time remained comparable (20–25 min). The T_g of the low-modulus cement was lower (75–78 °C) than that of the high-stiffness cement (103 °C). It was shown that sterilization of linoleic acid by autoclaving did not significantly affect the functional properties of low-modulus PMMA bone cement, making the component suitable for sterile production. Ultimately, the low-modulus cement exhibited handling and mechanical properties that more closely match those of osteoporotic vertebral bone with a screw holding capacity of under 2000 N, making it a promising alternative for use in combination with orthopedic hardware in applications where high-stiffness augmentation materials can result in undesired effects. Full article

In the field of rehabilitation and neuroscience, shape memory alloys play a crucial role as lightweight actuators. Devices are exploiting the shape memory effect by transforming heat into mechanical work. In rehabilitation applications, dynamic loading of the respective device occurs, which in turn influences the mechanical consequences of the phase transforming alloy. Hence in this work, dynamic thermomechanical material behavior of temperature-triggered phase transforming NiTi shape memory alloy (SMA) wires with different chemical compositions and geometries was experimentally investigated. Storage modulus and mechanical loss factor of NiTi alloys at different temperatures and loading frequencies were analyzed under force-controlled conditions. Counterintuitive storage modulus- and loss factor-dependent trends regarding the loading frequency dependency of the mechanical properties on the materials’ composition and geometry were, hence, obtained. It was revealed that loss factors showed a pronounced loading frequency dependency, whereas the storage modulus was not affected. It was shown that force-controlled conditions led to a lower storage modulus than expected. Furthermore, it turned out that a simple empirical relation could capture the characteristic temperature dependency of the storage modulus, which is an important input relation for modeling the rehabilitation device behavior under different dynamic and temperature loading conditions, taking directly into account the material behavior of the shape memory alloy. Full article

The surface topography of dental implants and micro-nano surface characterization have gained particular interest for the improvement of the osseointegration phases. The aim of this study was to evaluate the surface micro-nanomorphology and bioactivity (apatite forming ability) of Ossean^® surface, a resorbable blast medium (RBM) blasted surface further processed through the incorporation of a low amount of calcium phosphate. The implants were analyzed using environmental scanning electronic microscopy (ESEM), connected to Energy dispersive X-ray spectroscopy (EDX), field emission gun SEM-EDX (SEM-FEG) micro-Raman spectroscopy and X-ray photoelectron spectroscopy (XPS) before and after immersion in weekly refreshed Hank’s balanced salt solution (HBSS) for 28 days. The analysis of the samples before immersion showed a moderately rough surface, with micropits and microgrooves distributed on all of the surface; EDX microanalysis revealed the constitutional elements of the implant surface, namely titanium (Ti), aluminum (Al) and vanadium (V). Limited traces of calcium (Ca) and phosphorous (P) were detected, attributable to the incorporated calcium phosphate. No traces of calcium phosphate phases were detected by micro-Raman spectroscopy. ESEM analysis of the implant aged in HBSS for 28 days revealed a significantly different surface, compared to the implant before immersion. At original magnifications <2000×, a homogeneous mineral layer was present on all the surface, covering all the pits and microgrooves. At original magnifications ≥10,000×, the mineral layer revealed the presence of small microspherulites. The structure of these spherulites (approx. 2 µm diameter) was observed in nanoimmersion mode revealing a regular shape with a hairy-like contour. Micro-Raman analysis showed the presence of B-type carbonated apatite on the implant surface, which was further confirmed by XPS analysis. This implant showed a micro-nano-textured surface supporting the formation of a biocompatible apatite when immersed in HBSS. These properties may likely favor bone anchorage and healing by stimulation of mineralizing cells. Full article

Titanium and titanium alloys are promising implant metallic materials because of their high strengths, low elastic moduli, high corrosion resistances, and excellent biocompatibilities. A large difference in elastic modulus between the implant material and bone leads to a stress shielding effect, which increases the probability of implant separation or decrease in the bone density around it. Thus, a lower elastic modulus is required for a better implant metallic material. β titanium has a lower elastic modulus and high strength and can reduce the probability of the stress shielding effect. In this study, the applicability of the Ti–39Nb–6Zr+0.45Al alloy, obtained by adding a small amount of aluminum to the Ti–39Nb–6Zr alloy, as a biomedical implant material was evaluated. The mechanical properties and biocompatibility of the alloy were evaluated. The biocompatibility of Ti–39Nb–6Zr+0.45Al was similar to that of Ti–39Nb–6Zr according to in vitro and in vivo experiments. In addition, the biological corrosion resistances were evaluated through a corrosion test using a 0.9% NaCl solution, which is equivalent to physiological saline. The corrosion resistance was improved by the addition of Al. The yield strength of the Ti–39Nb–6Zr+0.45Al alloy was improved by approximately 20%. The excellent biocompatibility confirmed its feasibility for use as a biomedical implant material. Full article

Surface properties of composites such as roughness and color impact periodontal health and aesthetic outcomes. Novel bulk-fill composites with improved functionality are being introduced and, in light of the existing variety of finishing/polishing procedures, research of their surface properties is warranted. Sixty discs were prepared from bulk-fill composites (Filtek™ Bulk Fill Posterior Restorative and Fill-Up™) and incremental-fill Filtek™ Z250. They were further divided according to different polishing procedures (n = 5): three multi-step polishing procedures or finishing with a bur (control). Surface roughness (Ra) was measured using an atomic force microscope (The AFM Workshop TT-AFM). A spectrophotometer (Spectroshade Micro Optic) was used to determine color stability, after exposure to a coffee solution. Data were analyzed using two-way MANOVA (significance level of 5%). Resin composite type, polishing procedure, and their interaction had a statistically significant effect on surface roughness (p < 0.001) and color change (p < 0.001). Fill-Up™ exhibited the highest surface roughness and greatest color change. Differences in color change were statistically significant (p < 0.001). Filtek™ Bulk Fill registered the lowest surface roughness and color change, after the three-step polishing procedure. Both parameters were significantly correlated (ρ = 0.754, p < 0.001) and found to be material dependent and polishing-procedure dependent. Higher surface roughness relates to greater color changes. Full article