Tissue Engineering Applications Research Articles

Investigating the in-vitro bioactivity, biodegradability and drug release behavior of the newly developed PES/HA/WS biocompatible nanocomposites as bone graft substitute

The nucleation of carbonate-containing apatite on the biomaterials surface is regarded as a significant stage in bone healing process. In this regard, composites contained hydroxyapatite (Ca10(PO4)6(OH)2, HA), wollastonite (CaSiO3, WS) and polyethersulfone (PES) were synthesized via a simple solvent casting technique. The in-vitro bioactivity of the prepared composite films with different weight ratios of HA and WS was studied by placing the samples in the simulated body fluid (SBF) for 21 days. The results indicated that the the surface of composites containing 2 wt% HA and 4 wt% WS was completely covered by a thick bone-like apatite layer, which was characterized by Grazing incidence X-ray diffraction, attenuated total reflectance–Fourier transform infrared spectrometer, field emission electron microscopy and energy dispersive X-ray analyzer (EDX). The degradation study of the samples showed that the concentration of inorganic particles could not influence the degradability of the polymeric matrix, where all samples expressed similar dexamethasone (DEX) release behavior. Moreover, the in-vitro cytotoxicity results indicated the significant cyto-compatibility of all specimens. Therefore, these findings revealed that the prepared composite films composed of PES, HA, WS and DEX could be regarded as promising bioactive candidates with low degradation rate for bone tissue engineering applications.

Chromatin Condensation Delays Senescence in Human Mesenchymal Stem Cells by Safeguarding Nuclear Damages during In Vitro Expansion

Human mesenchymal stem cells (hMSCs) are multipotent cells that differentiate into adipocytes, chondrocytes, and osteoblasts. Owing to their differentiation potential, hMSCs are among the cells most frequently used for therapeutic applications in tissue engineering and regenerative medicine. However, the number of cells obtained through isolation alone is insufficient for hMSC-based therapies and basic research, which necessitates in vitro expansion. Conventionally, this is often performed on rigid surfaces such as tissue culture plates (TCPs). However, during in vitro expansion, hMSCs lose their proliferative ability and multilineage differentiation potential, rendering them unsuitable for clinical use. Although multiple approaches have been attempted to maintain hMSC stemness during prolonged expansion, finding a suitable culture system remains an unmet need. Recently, a few research groups have shown that hMSCs maintain their stemness over long passages when cultured on soft substrates. In addition, it has been shown that hMSCs cultured on soft substrates have more condensed chromatin and lower levels of histone acetylation compared to those cultured on stiff substrates. Furthermore, it has also been shown that condensing/decondensing chromatin by deacetylation/acetylation can delay replicative senescence in hMSCs during long-term expansion on TCPs. However, the mechanism by which chromatin condensation/decondensation influences nuclear morphology and DNA damage, which are strongly related to the onset of senescence, remains unknown. To answer this question, we cultured hMSCs for long duration in the presence of epigenetic modifiers, histone acetyltransferase inhibitor (HATi), which promotes chromatin condensation by preventing histone acetylation, and histone deacetylase inhibitor (HDACi), which promotes chromatin decondensation, and investigated their effects on various nuclear markers related to senescence. We found that consistent acetylation causes severe nuclear abnormalities, whereas chromatin condensation by deacetylation helps to safeguard the nucleus from damage caused by in vitro expansion.

A Mesenchymal stem cell Aging Framework, from Mechanisms to Strategies.

Mesenchymal stem cells (MSCs) are extensively researched for therapeutic applications in tissue engineering and show significant potential for clinical use. Intrinsic or extrinsic factors causing senescence may lead to reduced proliferation, aberrant differentiation, weakened immunoregulation, and increased inflammation, ultimately limiting the potential of MSCs. It is crucial to comprehend the molecular pathways and internal processes responsible for the decline in MSC function due to senescence in order to devise innovative approaches for rejuvenating senescent MSCs and enhancing MSC treatment. We investigate the main molecular processes involved in senescence, aiming to provide a thorough understanding of senescence-related issues in MSCs. Additionally, we analyze the most recent advancements in cutting-edge approaches to combat MSC senescence based on current research. We are curious whether the aging process of stem cells results in a permanent "memory" and if cellular reprogramming may potentially revert the aging epigenome to a more youthful state.

Shear Flow-Assembled Janus Membrane with Bifunctional Osteogenic and Antibacterial Effects for Guided Bone Regeneration.

Guided bone regeneration (GBR) membranes that reside at the interface between the bone and soft tissues for bone repair attract increasing attention, but currently developed GBR membranes suffer from relatively poor osteogenic and antibacterial effects as well as limited mechanical property and biodegradability. We present here the design and fabrication of a bifunctional Janus GBR membrane based on a shear flow-driven layer by a layer self-assembly approach. The Janus GBR membrane comprises a calcium phosphate-collagen/polyethylene glycol (CaP@COL/PEG) layer and a chitosan/poly(acrylic acid) (CHI/PAA) layer on different sides of a collagen membrane to form a sandwich structure. The membrane exhibits good mechanical stability and tailored biodegradability. It is found that the CaP@COL/PEG layer and CHI/PAA layer contribute to the osteogenic differentiation and antibacterial function, respectively. In comparison with the control group, the Janus GBR membrane displays a 2.52-time and 1.84-time enhancement in respective volume and density of newly generated bone. The greatly improved bone repair ability of the Janus GBR membrane is further confirmed through histological analysis, and it has great potential for practical applications in bone tissue engineering.

Reprogramming tendon healing: a guide to novel molecular tools

Tendons are a frequent site of injury, which greatly impairs the movement and locomotion of patients. Regrettably, injuries at the tendon frequently require surgical intervention, which leads to a long path to recovery. Moreover, the healing of tendons often involves the formation of scar tissue at the site of injury with poor mechanical properties and prone to re-injury. Tissue engineering carries the promise of better and more effective solutions to the improper healing of tendons. Lately, the field of regenerative medicine has seen a significant increase in the focus on the potential use of non-coding RNAs (e.g., siRNAs, miRNAs, and lncRNAs) as molecular tools for tendon tissue engineering. This class of molecules is being investigated due to their ability to act as epigenetic regulators of gene expression and protein production. Thus, providing a molecular instrument to fine-tune, reprogram, and modulate the processes of tendon differentiation, healing, and regeneration. This review focuses particularly on the latest advances involving the use of siRNAs, miRNAs, and lncRNAs in tendon tissue engineering applications.

A comparative study of 3D-printed scaffolds fabricated from different hydroxyapatite sources for bone tissue engineering applications

As a naturally occurring mineral component of bone, hydroxyapatite (HA) is a preferred bone-repair material in clinical medicine. This adaptable biomaterial is developed from synthetic or biological sources. The goal of this study was to investigate the physicochemical and biological properties of scaffolds made from diverse hydroxyapatite sources. We prepared hydroxyapatite powders either from naturally occurring sources, using decellularized bone extracellular matrix (DBECM) and deproteinized bovine bone (DBB), or in the laboratory using the co-precipitation method. Although XRD analysis confirmed the pure phase of HA in all three powders, XRF data found some other trace elements in bone-derived biomaterials, such as magnesium and strontium. Further, FTIR spectra showed DBECM-associated macromolecule peaks. In the second phase of this study, the particles were mixed with sodium alginate to produce bioink for 3D printing via extrusion. SEM photographs of the printed scaffolds showed a grid-like porous structure with large, interconnected pores. Although all of the scaffolds retained their structural integrity after 28 days submerged in the culture medium, DBECM-based scaffolds degraded the most. The test of Young’s modulus and compressive strength of scaffolds revealed that all scaffolds are strong enough to be employed in non-weight-bearing applications. The biological characteristics of the scaffolds were then evaluated via cytotoxicity analysis, SEM examination of cell attachment, MTT assay, ALP activity assessment, immunofluorescence staining, and Alizarin red staining. The results indicated that all three scaffolds provided a surface that promoted adhesion and proliferation of rat bone marrow mesenchymal stem cells (rMSCs), as well as stimulated the production of mineralized extracellular matrices. Additionally, our findings showed that scaffolds made from DBECM powders offer the best environment for cell activity. This is most likely due to the presence of trace elements and protein macromolecules in DBECM powders that are not found in DBB and synthetic HA. In conclusion, our comparative study revealed that while all three scaffolds are suitable for bone tissue engineering, DBECM-based scaffolds outperform DBB and synthetic HA.

Research progress on the application of magnesium phosphate bone cement in bone defect repair: A review.

Bone defects arising from diverse causes, such as traffic accidents, contemporary weapon usage, and bone-related disorders, present significant challenges in clinical treatment. Prolonged treatment cycles for bone defects can result in complications, impacting patients' overall quality of life. Efficient and timely repair of bone defects is thus a critical concern in clinical practice. This study aims to assess the scientific progress and achievements of magnesium phosphate bone cement (MPC) as an artificial bone substitute material. Additionally, the research seeks to explore the future development path and clinical potential of MPC bone cement in addressing challenges associated with bone defects. The study comprehensively reviews MPC's performance, encompassing e.g. mechanical properties, biocompatibility, porosity, adhesion and injectability. Various modifiers are also considered to broaden MPC's applications in bone tissue engineering, emphasizing drug-loading performance and antibacterial capabilities, which meet clinical diversification requirements. In comparison to alternatives such as autogenous bone transplantation, allograft, polymethyl methacrylate (PMMA), and calcium phosphate cement (CPC), MPC emerges as a promising solution for bone defects. It addresses limitations associated with these alternatives, such as immunological rejection and long-term harm to patients. MPC can control heat release during the curing process, exhibits superior mechanical strength, and has the capacity to stimulate new bone growth. MPC stands out as an artificial bone substitute with appropriate mechanical strength, rapid degradation, non-toxicity, and good biocompatibility, facilitating bone repair and regeneration. Modification agents can enhance its clinical versatility. Future research should delve into its mechanical properties and formulations, expanding clinical applications to create higher-performing and more medically valuable alternatives in bone defect repair.

The “Forgotten” Hydroxyapatite Crystals in Regenerative Bone Tissue Engineering: A Critical Review

Bone regeneration using Bone Morphogenetic Proteins (BMPs) alongside various engineered scaffolds has attracted considerable attention over the years. The field has seen extensive research in preclinical animal models, leading to the approval of two products and guiding the quest for new materials. Natural and synthetic polymers, ceramics, and composites have been used to fabricate the necessary porous 3D scaffolds and delivery systems for BMPs. Interestingly, all reported applications in the literature are triumphant. Evaluation of the results is typically based on histologic assessment after appropriate staining and radiological modalities, providing morphological identification of the newly formed bone and describing cells and the organic compound. Remarkably, while these evaluation methods illustrate mineralization, they are not capable of identifying hydroxyapatite crystals, the mineral component of the bone, which are crucial for its mechanical properties, structure, integrity, and long-term stability of regenerated bone tissue. This review aims to focus on the different scaffolds used in bone tissue engineering applications and underline the pressing need for techniques that could recognize the presence of hydroxyapatite crystals as well as their characteristics in bone tissue engineering, which will provide a more complete and comprehensive assessment of the successful results.

Biomimetic Approach for Enhanced Mechanical Properties and Stability of Self-Mineralized Calcium Phosphate Dibasic–Sodium Alginate–Gelatine Hydrogel as Bone Replacement and Structural Building Material

Mineralized materials are gaining increased interest recently in a number of fields, especially in bone tissue engineering as bone replacement materials as well as in the architecture-built environment as structural building materials. Until the moment, there has not been a unified sustainable approach that addresses this multi-scale application objective by developing a self-mineralized material with minimum consumption of materials and processes. Thus, in the current study, a hydrogel developed from sodium alginate, gelatine, and calcium phosphate dibasic (CPDB) was optimized in terms of rheological properties and mineralization capacity through the formation of hydroxyapatite crystals. The hydrogel composition process adopted a three-stage, thermally induced chemical cross-linking to achieve a stable and enhanced hydrogel. The 6% CPDB-modified SA–gelatine hydrogel achieved the best rheological properties in terms of elasticity and hardness. Different concentrations of epigallocatechin gallate were tested as well as a rheological enhancer to optimize the hydrogel and to boost its anti-microbial properties. However, the results from the addition of EPGCG were not considered significant; thus, the 6% CPDB-modified SA–gelatine hydrogel was further tested for mineralization by incubation in various media, without and with cells, for 7 and 14 days, respectively, using scanning electron microscopy. The results revealed significantly enhanced mineralization of the hydrogel by forming hydroxyapatite platelets of the air-incubated hydrogel (without cells) in non-sterile conditions, exhibiting antimicrobial properties as well. Similarly, the air-incubated bioink with osteosarcoma SaOs-2 cells exhibited dense mineralized topology with hydroxyapatite crystals in the form of faceted spheres. Finally, the FBS-incubated hydrogel and FBS-incubated bioink, incubated for 7 and 14 days, respectively, exhibited less densely mineralized topology and less distribution of the hydroxyapatite crystals. The degradation rate of the hydrogel and bioink incubated in FBS after 14 days was determined by the increase in dimensions of the 3D-printed samples, which was between 5 to 20%, with increase in the bioink samples dimensions in comparison to their dimensions post cross-linking. Meanwhile, after 14 days, the hydrogel and bioink samples incubated in air exhibited shrinkage: a 2% decrease in the dimensions of the 3D-printed samples in comparison to their dimensions post cross-linking. The results prove the capacity of the developed hydrogel in achieving mineralized material with anti-microbial properties and a slow-to-moderate degradation rate for application in bone tissue engineering as well as in the built environment as a structural material using a sustainable approach.

Microfluidic Platforms to Screen Granular Hydrogel Microenvironments for Tissue Regeneration

AbstractGranular hydrogels have emerged as a promising class of biomaterials in medical research, enabling independent control of matrix stiffness within a porous biomaterial. Such microgel packings comprise interconnected pores and high surface‐to‐volume ratios. These features facilitate cell viability and nutrient diffusion, which are critical in enabling tissue regeneration. Despite the current interest in granular hydrogels for tissue engineering applications, only a few in vitro platforms are used to investigate cell interactions, limiting their design, and translation. In this study, microfluidic platforms able to reproducibly confine and immobilize microgels without the need for secondary cross‐linking are developed. Protocols are established for the generation of human bone marrow‐derived mesenchymal stem/stromal cells (hBM‐MSC)‐infiltrated microporous substrates and early‐time responses of cells to their environment are studied. Further, a tissue invasion assay is established, where cells infiltrate granular materials at different rates depending on growth factor presence or material properties. This platform is compatible with a range of different granular materials, and it is envisioned to have significant utility as a pre‐clinical tool for the rational design of materials for tissue healing applications.

3D Printing of Succulent‐Inspired Microneedle Array for Enhanced Tissue Adhesion and Controllable Drug Release

AbstractControllable and long‐term release remains a great challenge in current drug delivery systems. Benefiting from their efficient drug loading and painless administration, microneedles (MNs) have emerged as a promising platform for transdermal drug delivery, while they often fail to achieve long‐term tissue adhesion and controllable extended drug release. Here, 3D printing of an innovative MN patch is presented with succulent‐inspired responsive microstructures and light‐controllable long‐term release capability. The MN exhibits a reversible shrink‐swell volume change behavior in response to surrounding humidity, which enables sufficient mechanical strength for skin penetration under the shrinkage conditions and efficient long‐term adhesion when swollen in skin tissues. Moreover, the MN patch introduces a controllable long‐term drug release system, achieved through the integration of thiolated heparin (Hep‐SH) for sustained growth factor release and graphene oxide (GO) nanosheets for controlled drug release via near infrared (NIR) laser irradiation. The MN patches with growth factor loading have good biocompatibility and can promote the proliferation, migration, and proangiogenesis of endothelial cells is further demonstrated. Thus, it is believed that such flexible MN patches can be promising candidates for controllable long‐term transdermal drug delivery as well as other related tissue engineering applications.

Biologically active sodium pentaborate pentahydrate and Hypericum perforatum oil loaded poly vinyl alcohol: chitosan membranes

In this study, sodium pentaborate pentahydrate (NaB) and Hypericum perforatum (HP) oil were incorporated into polyvinyl alcohol (PVA) and chitosan (CH) polymer blend to obtain membranes by solution casting method. In order to see the synergistic effects of NaB and HP oil on the biological and physical properties of the membranes NaB and HP oil were incorporated into membrane matrix in different ratios. Fourier-transform infrared spectroscopy (FTIR) results showed that no significant bond formation between the bioactive components and the PVA:CH matrix. According to mechanical test results, Young's Modulus and elongation at break decreased from 426 MPa to 346 MPa and 52.23 % to 15.11 % for neat PVA:CH membranes and NaB and HP oil incorporated PVA:CH (PVA:CH@35NaB:HP) membranes, respectively. Antimicrobial activity tests have shown the membranes were over 99 % effective against Escherichia coli, Staphylococcus aureus, and Candida albicans, underlining their potential for infection control. Cytocompatibility assay performed with Human Dermal Fibroblast (HDFa) cells highlight the biocompatibility of the membranes, revealing 74.84 % cell viability after 72 h. The properties of NaB and HP oil doped PVA:CH based membranes obtained from these experiments reveal the promise of a versatile membrane for applications in wound healing, tissue engineering and other biomedical fields.

Elderberry (Sambucus nigra) and hawthorn (Crataegus oxyacantha) extract additives in carboxymethyl chitosan scaffolds for osteochondral tissue engineering applications

Elderberry (<i>Sambucus nigra</i>) and hawthorn (<i>Crataegus oxyacantha</i>) extract additives in carboxymethyl chitosan scaffolds for osteochondral tissue engineering applications

Exploring self-crosslinked hydrogel: Carboxymethyl chitosan and oxidized alginate for wound healing applications

This study reports the development and evaluation of self-healing hydrogel composed of carboxymethyl chitosan (CmCh) and oxidized alginate, with a focus on wound healing applications. Oxidized alginate was synthesized via metaperiodate oxidation and characterised. The hydrogels were formulated using Schiff base reactions with two weight ratios of oxidized alginate and CmCh, and their cross-linking mechanism was proposed. The developed hydrogels were characterised for their gelation, pH, self-healing capability, conductivity, and injectability. The optimized hydrogel was further evaluated for antibacterial activity, cytotoxicity, haemocompatibility, and in vitro wound healing studies. Results revealed that hydrogels with the lowest ratio of oxidized alginate to CmCh exhibited superior gelation, tensile strength, and injectability without needle clogging. Additionally, rapid self-healing of hydrogels was confirmed via Scanning Electron Microscope (SEM) analysis. The optimized hydrogel displayed good antibacterial activity against Staphylococcus aureus and Escherichia coli, alongside haemocompatibility. Additionally, in vivo studies demonstrated comparable wound healing efficacy to marketed formulations and significant improvement over control groups. These findings indicates the potential of self-healing hydrogels made up of oxidized alginate and CmCh for tissue engineering applications.

Alendronate releasing silk fibroin 3D bioprinted scaffolds for application in bone tissue engineering: Effects of alginate concentration on printability, mechanical properties and stability

3D bioprinting uses biomaterials combined with cells to develop living constructs. This study explores the optimization of natural polymers, including silk fibroin, gelatin, and alginate, as bioink for 3D bioprinting in bone tissue engineering. The physicochemical properties of the bioink were thoroughly examined, revealing high water uptake and reduced degradation rate due to the addition of silk fibroin. The compressive modulus increased with higher alginate concentration. Rheological analysis confirmed shear-thinning properties and viscoelasticity of the ink. Through meticulous parameter optimization, the ink achieved the highest print accuracy with 4 % w/v alginate content. The printed scaffolds exhibited both macro and micro porosity, making them suitable for bone tissue regeneration. Furthermore, the scaffolds remained stable in culture medium for 36 days. The optimal composition for the hydrogel was determined to be a blend of 5 % fibroin, 7 % gelatin, and 4 % alginate in equal ratios. The bioink demonstrated excellent biocompatibility and, when supplemented with alendronate, enhanced alkaline phosphatase activity in MG-63 osteoblast-like cells. This finding indicates the commitment of cells toward the osteoblastic phenotype. Overall, this study successfully optimized the bioink and bioprinting process for bone tissue engineering applications, highlighting its promising potential for future advancements in the field.

Electrospun nanostructured heparin conjugated-poly-ε-caprolactone based scaffold promote differentiation of smooth muscle cells from adipose mesenchymal stem cells

BackgroundTissue engineering integrating cell-based approaches is promising for urinary tract injury treatments. PurposeExploring the Impact of Nanostructured Heparin Conjugated-Polycaprolactone (HPCL) on the differentiation of Adipose Mesenchymal Stem Cells (ADMSCs) into Smooth Muscle Cells (SMCs). MethodsPCL nanofibers were produced by electrospinning and conjugated with heparin. The surface properties of the scaffolds were analyzed using FE-SEM for imaging, FTIR for investigation of chemical structure, assessments of surface roughness, thermomechanical characteristics, and the contact angle to evaluate wettability. The study included six groups; Group I: ADMSCs cells, Group II: ADMSCs seeded on PCL, Group III: ADMSCs seeded on HPCL, Group IV: SMCs cells, Group V: SMCs seeded on PCL, and Group VI: MSCs seeded on HPCL. SMCs differentiation was evaluated using specific genes and proteins. ResultsImmobilization of heparin was confirmed by amide I (3427 cm−1) and it was significantly raised on HPCL scaffolds. SMCs + HPCL group revealed the highest cell viability and the most significant increase in differentiation-related genes and proteins at 7 days (p < 0.05). ConclusionHPCL nanofibers have an effect on SMCs differentiation, which are the functional units of the ureter. Therefore, they could represent a valuable tool for tissue engineering applications in ureter repair.

3D Bioprinting of Sugar Beet Pectin through Horseradish Peroxidase-Catalyzed Cross-Linking.

Horseradish peroxidase (HRP)-mediated hydrogelation, caused by the cross-linking of phenolic groups in polymers in the presence of hydrogen peroxide (H2O2), is an effective route for bioink solidification in 3D bioprinting. Sugar beet pectin (SBP) naturally has cross-linkable phenols through the enzymatic reaction. Therefore, chemical modifications are not required, unlike the various polymers that have been used in the enzymatic cross-linking system. In this study, we report the application of SBP in extrusion-based bioprinting including HRP-mediated bioink solidification. In this system, H2O2 necessary for the solidification of inks is supplied in the gas phase. Cell-laden liver lobule-like constructs could be fabricated using bioinks consisting of 10 U/mL HRP, 4.0 and 6.0 w/v% SBP, and 6.0 × 106 cells/mL human hepatoblastoma (HepG2) cells exposed to air containing 16 ppm of H2O2 concurrently during printing and 10 min postprinting. The HepG2 cells enclosed in the printed constructs maintained their viability, metabolic activity, and hepatic functions from day 1 to day 7 of the culture, which indicates the cytocompatibility of this system. Taken together, this result demonstrates the potential of SBP and HRP cross-linking systems for 3D bioprinting, which can be applied in tissue engineering applications.

Hydrogen Sulfide Delivery to Enhance Bone Tissue Engineering Cell Survival

Though crucial for natural bone healing, local calcium ion (Ca2+) and phosphate ion (Pi) concentrations can exceed the cytotoxic limit leading to mitochondrial overload, oxidative stress, and cell death. For bone tissue engineering applications, H2S can be employed as a cytoprotective molecule to enhance mesenchymal stem cell (MSC) tolerance to cytotoxic Ca2+/Pi concentrations. Varied concentrations of sodium hydrogen sulfide (NaSH), a fast-releasing H2S donor, were applied to assess the influence of H2S on MSC proliferation. The results suggested a toxicity limit of 4 mM for NaSH and that 1 mM of NaSH could improve cell proliferation and differentiation in the presence of cytotoxic levels of Ca2+ (32 mM) and/or Pi (16 mM). To controllably deliver H2S over time, a novel donor molecule (thioglutamic acid—GluSH) was synthesized and evaluated for its H2S release profile. Excitingly, GluSH successfully maintained cytoprotective level of H2S over 7 days. Furthermore, MSCs exposed to cytotoxic Ca2+/Pi concentrations in the presence of GluSH were able to thrive and differentiate into osteoblasts. These findings suggest that the incorporation of a sustained H2S donor such as GluSH into CaP-based bone graft substitutes can facilitate considerable cytoprotection, making it an attractive option for complex bone regenerative engineering applications.

Enhanced Electroactive Phases of Poly(vinylidene Fluoride) Fibers for Tissue Engineering Applications.

Nanofibrous materials generated through electrospinning have gained significant attention in tissue regeneration, particularly in the domain of bone reconstruction. There is high interest in designing a material resembling bone tissue, and many scientists are trying to create materials applicable to bone tissue engineering with piezoelectricity similar to bone. One of the prospective candidates is highly piezoelectric poly(vinylidene fluoride) (PVDF), which was used for fibrous scaffold formation by electrospinning. In this study, we focused on the effect of PVDF molecular weight (180,000 g/mol and 530,000 g/mol) and process parameters, such as the rotational speed of the collector, applied voltage, and solution flow rate on the properties of the final scaffold. Fourier Transform Infrared Spectroscopy allows for determining the effect of molecular weight and processing parameters on the content of the electroactive phases. It can be concluded that the higher molecular weight of the PVDF and higher collector rotational speed increase nanofibers' diameter, electroactive phase content, and piezoelectric coefficient. Various electrospinning parameters showed changes in electroactive phase content with the maximum at the applied voltage of 22 kV and flow rate of 0.8 mL/h. Moreover, the cytocompatibility of the scaffolds was confirmed in the culture of human adipose-derived stromal cells with known potential for osteogenic differentiation. Based on the results obtained, it can be concluded that PVDF scaffolds may be taken into account as a tool in bone tissue engineering and are worth further investigation.

Physicochemical and Biological Characterization of Propolis-Loaded Composite Polyamide-6/Soybean Protein Nanofibers for Wound Healing Applications

Dressings are pivotal in aiding wound recovery, yet they are burdened by significant constraints. In addition to possessing antibacterial properties, the effectiveness of a wound dressing must be rigorously assessed for its potential utility across diverse medical applications. Propolis (Pro) is widely acknowledged for its antioxidant, antibacterial, and anti-inflammatory properties, positioning it as a promising candidate for wound healing and tissue engineering applications. This study presents a novel composite nanofiber (NF) material comprising polyamide-6 (PA6) and soybean protein as matrix components, incorporating Pro as a payload. We aimed to mimic the extracellular matrix, thus promotion the proliferation and adhesion of mouse embryonic fibroblast cells (NIH3T3) on the scaffold's surface. The morphology and composition of the NFs, examined through scanning electron microscopy (SEM) imaging and Fourier-transform infrared (FTIR) spectroscopy. The Pro-loaded fibers exhibited diameters ranging from 102 to 270nm. Notably, the NFs demonstrated a dose-dependent increase in porosity, water vapor transmission rate, and fluid uptake capacity. However, the hydrophilic categorized nanofibers (NFs) did not exhibit a concentration-dependent effect on contact angle measurements. After a 15-day period, the Pro-loaded fibers experienced degradation within the range of 39-53%. Using UV-visible spectrophotometry, the amount of pro released was measured. The rates of release were found to be 29.93%, 41.02%, and 60.06% within 24hours for PA6/Soy/Pro5%, PA6/Soy/Pro10%, and PA6/Soy/Pro20% NFs, respectively. Importantly, our study showcased the significant inhibitory effects of Pro-loaded fibers on the growth of E. coli and S. aureus, with inhibition increasing proportionally with higher Pro concentrations. This observation underscores the potential of Pro-based composite NFs as effective antimicrobial agents for wound dressings and tissue scaffolds. Furthermore, the biocompatibility of the fabricated nanofibrous mats was confirmed through the MTT test, affirming the non-toxic nature of all formulations. SEM imaging further demonstrated the optimal adhesion of cells on the NFs, confirming the findings of the MTT test and highlighting the suitability of Pro-loaded NFs for cell attachment and proliferation.