Polycaprolactone Surface Research Articles

The role of lignin as interfacial compatibilizer in designing lignocellulosic-polyester composite films

Advancing nanocomposites requires a deep understanding and careful design of nanoscale interfaces, as interfacial interactions and adhesion significantly influence the physical and mechanical properties of these materials. This study demonstrates the effectiveness of lignin nanoparticles (LNPs) as interfacial compatibilizer between hydrophilic cellulose nanofibrils (CNF) and a hydrophobic polyester, polycaprolactone (PCL). In this context, we conducted a detailed analysis of surface-to-bulk interactions in both wet and dry conditions using advanced techniques such as quartz crystal microbalance with dissipation (QCM-D), atomic force microscopy (AFM), water contact angle (WCA) measurements, broadband dielectric spectroscopy (BDS), and inverse gas chromatography (IGC).QCM-D was employed to quantify the adsorption behavior of LNPs on CNF and PCL surfaces, demonstrating LNPs’ capability to interact with both hydrophilic and hydrophobic phases, thereby enhancing composite material properties. LNPs showed extensive adsorption on a CNF model film (1186 ± 178 ng.cm−2) and a lower but still significant adsorption on a PCL model film (270 ± 64 ng.cm−2). In contrast, CNF adsorption on a PCL model film was the lowest, with a sensed mass of only 136 ± 35 ng.cm−2. These findings were further supported by comparing the morphology and wettability of the films before and after adsorption, using AFM and WCA analyses. Then, to gain insights into the molecular-level interactions and molecular mobility within the composite in dry state, BDS was employed. The BDS results showed that LNPs improved the dispersion of PCL within the CNF network. To further investigate the impact of LNPs on the composites’ interfacial properties, IGC was employed. This analysis showed that the composite films containing LNPs exhibited lower surface energy compared to those composed of only CNF and PCL. The presence of LNPs likely reduced the availability of surface hydroxyl groups, thus modifying the physicochemical properties of the interface. These changes were particularly evident in the heterogeneity of the surface energy profile, indicating that LNPs significantly altered the interfacial characteristics of the composite materials.Overall, these findings emphasize the necessity to control the interfaces between components for next-generation nanocomposite materials across diverse applications.

Polycaprolactone scaffold surface modification with soft X-ray/extreme ultraviolet (SXR/EUV) radiation and low-temperature oxygen and nitrogen plasma for biomedical applications

Modification of the surfaces of polymeric scaffolds is often required to make the material suitable for specific tissue engineering applications. Physico-chemical properties of scaffolds can be altered using various methods, such as plasma treatment, laser processing, chemical modifications, grafting with nanoparticles, or surface coating. In this paper physico-chemical modification of polycaprolactone (PCL) surface fibers was performed by exposing PCL samples to simultaneous soft X-ray/extreme ultraviolet (SXR/EUV) radiation and low-temperature, SXR/EUV-induced, nitrogen, and oxygen plasmas. The physical and chemical changes on modified PCL surfaces were examined using a scanning electron microscope and X-ray photoelectron spectroscopy, respectively. The effects of physico-chemical scaffold surface changes were verified with biological tests, i.e., MTT assay and immunofluorescence on murine osteoblast cell line (7F2). It was found that exposure of scaffolds to ionizing radiation and low-temperature plasmas induced strong chemical changes on their surface, i.e., appearance of various new chemical groups. Also, smoothing of the surface of PCL fibers, i.e., disappearance or significant reduction of the size of micropores on their fibers was also observed. Increased viability and adhesion of 7F2 osteoblasts on modified PCL samples after 24 h cell culture compared to non-treated PCL was also confirmed.Graphical abstract

Regulation of mitochondrial network architecture and function in mesenchymal stem cells by micropatterned surfaces

Abstract Mitochondrial network architecture, which is closely related to mitochondrial function, is mechanically sensitive and regulated by multiple stimuli. However, the effects of microtopographic cues on mitochondria remain poorly defined. Herein, polycaprolactone (PCL) surfaces were used as models to investigate how micropatterns regulate mitochondrial network architecture and function in rat adipose-derived stem cells (rASCs). It was found that large pit (LP) induced rASCs to form larger and more complex mitochondrial networks. Consistently, the expression of key genes related to mitochondrial dynamics revealed that mitochondrial fusion (MFN1 and MFN2) and midzone fission (DRP1 and MFF) were increased in rASCs on LP. In contrast, the middle pit (MP) enhanced mitochondrial biogenesis, as evidenced by the larger mitochondrial area and higher expression of PGC-1. Both LP and MP promoted ATP production in rASCs. It is likely that LP increased ATP levels through modulating mitochondrial network architecture while MP stimulated mitochondria biogenesis to do so. Our study clarified the regulation of micropatterned surfaces on mitochondria, highlighting the potential of LP and MP as a simple platform to stimulate mitochondria and the subsequent cellular function of MSCs.

Non-invasive Thermohydrodynamic Approach for Fast Cell Manipulation at the Microscale

Thermal gradients have emerged as a promising technique for manipulating and sorting biological material at the microscale, holding considerable potential in lab-on-a-chip technology. Herein, we propose a non-invasive thermohydrodynamic approach for fast cell manipulation using a microfluidic open-to-air device. Cell discrimination is achieved by simply changing the temperature gradient toward the control of the convective effect on their displacement. First, the size and morphology/roughness-based motion capabilities were modeled using polystyrene (PS) microparticles with different sizes (5 and 20 μm) and polycaprolactone (PCL) microspheres, respectively. Computational fluid dynamics simulations of the generated flow were also carried out to demonstrate the influence of both the thermohydrodynamic and Marangoni effects in the PS particle displacement, where the thermally induced convective effect was not enough to move the microparticles inside the channel, but the combination of thermally induced convection together with the Marangoni effect. Indeed, small particles (5 μm) followed a full convective path, whereas the bigger ones (20 μm) exhibited a rolling motion on the substrate from the cold side to the hot side. Also, the relationship between in-flow speed and PCL (≈ 20 μm) surface roughness confirmed the driving force of this convection-based approach. Then, the microfluidic device was successfully used to separate Henrietta Lacks cancer cells (HeLa) from red blood (RBCs) and fibroblast (HFF-1) cells. To this end, thermal gradients were tailored to achieve the desired thermohydrodynamic effect, showing a highly versatile performance. Both cell models (HeLa-RBCs and HeLa-HFF-1), due to rationale tweaking of the imposed temperature gradients (ΔT = 10 K, 303–293 K, and ΔT = 5 K, 303–298 K), were efficiently separated in less than 5 and 60 s, respectively; with excellent cell viabilities. The proposed microfluidic approach holds considerable promise for thermohydrodynamic sorting and manipulation of biological material by non-invasive methods using portable instrumentation. The potential parallelization of the thermal-convective approach opens new avenues for early disease diagnosis (liquid biopsies) or the study of biological systems, even at physiological temperatures with a potential impact in cell (organ)-on-a-chip technologies.

Characterization of bacterial biofilms developed on the biodegradable polylactide and polycaprolactone polymers containing birch tar in an aquatic environment

Birch tar was added to polylactide (PLA) and polycaprolactone (PCL) to create films with antimicrobial properties. After incubating the films for seven days in lake water, the diversity of bacterial communities developed on the surfaces of PCL and PLA with embedded birch tar (1 %, 5 %, and 10 %, w/w) was assessed with amplicon sequencing of the 16S rRNA gene on a MiSeq platform (Illumina). Notably, Aquabacterium and Caulobacter were more abundant at the surface of PCL compared to PLA (13.4 % vs 0.2 %, p < 0.001 and 9.5 % vs 0.2 %, p < 0.001, respectively) while Hydrogenophaga was significantly more abundant at the surface of PLA compared to PCL (6.1 % vs 1.8 %, p < 0.01). Overall, lower birch tar concentrations (1 % and 5 % on both polymers) stimulated bacterial diversity in biofilms compared to the control. The number of reeds assigned to Flavobacterium and Aquabacterium showed a rising trend with the increase of birch tar concentration on the surface of both polymers.

Modelling the development of biological structures displaying longitudinal geometries in vitro: culturing pluripotent stem cells on plasma-treated, growth factor-coupled polycaprolactone fibres

Many biological structures such as nerves, blood and lymphatic vessels, and muscle fibres exhibit longitudinal geometries with distinct cell types extending along both the length and width of internal linear axes. Modelling these three-dimensional structures in vitro is challenging: the best-defined stem-cell differentiation systems are monolayer cultures or organoids using pluripotent stem cells. Pluripotent stem cells can differentiate into functionally mature cells depending on the signals received, holding great promise for regenerative medicine. However, the integration of in vitro differentiated cell types into diseased tissue remains a challenge. Engineered scaffolds can bridge this gap if the appropriate signalling systems are incorporated into the scaffold. Here, we have taken a biomimicry approach to generate longitudinal structures in vitro. In this approach, mouse embryonic stem cells are directed to differentiate to specific cell types on the surface of polycaprolactone (PCL) fibres treated by plasma-immersion ion implantation and to which with lineage-specifying molecules have been covalently immobilised. We demonstrate the simplicity and utility of our method for efficiently generating high yields of the following cell types from these pluripotent stem cells: neurons, vascular endothelial cells, osteoclasts, adipocytes, and cells of the erythroid, myeloid, and lymphoid lineages. Strategically arranged plasma-treated scaffolds with differentiated cell types could ultimately serve as a means for the repair or treatment of diseased or damaged tissue.

Natural bone inspired core–shell triple-layered gel/PCL/gel 3D printed scaffolds for bone tissue engineering

Despite technological advancements in bone tissue engineering, it is still a challenge to fabricate a scaffold with high bioactivity as well as high mechanical strength that can promote osteogenesis as well as bear load. Here we developed a 3D printed gel-polymer multi-layered hybrid scaffold. The innermost layer is porous gel-based framework made of gelatin/carboxymethyl-chitin/nano-hydroxyapatite and is cryogenically 3D printed. Further, the second and middle layer of micro-engineered polycaprolactone (PCL) is infused in the gel with controlled penetration and tuneable coating thickness. The PCL surface is further coated with a third and final thin layer of gel matrix used for the first layer. This triple-layered structure demonstrates compression strength and modulus of 13.07 ± 1.15 MPa and 21.8 ± 0.82 MPa, respectively, post 8 weeks degradation which is >3000% and >700% than gel scaffold. It also shows degradation of 6.84 ± 0.70% (83% reduction than gel scaffold) after 12 weeks and swelling of 69.09 ± 6.83% (81% reduction) as compared to gel scaffolds. Further, nearly 300%, 250%, 50%, and 440% increase in cellular attachment, proliferation, protein generation, and mineralization, respectively are achieved as compared to only PCL scaffolds. Thus, these hybrid scaffolds offer high mechanical strength, slow degradation rate, high bioactivity, and high osteoconductivity. These multifunctional scaffolds have potential for reconstructing non-load-bearing bone defects like sinus lift, jaw cysts, and moderate load-bearing like reconstructing hard palate, orbital palate, and other craniomaxillofacial bone defects.

Characterization of the Protein Corona of Three Chairside Hemoderivatives on Melt Electrowritten Polycaprolactone Scaffolds.

In tissue engineering, the relationship between a biomaterial surface and the host's immune response during wound healing is crucial for tissue regeneration. Despite hemoderivative functionalization of biomaterials becoming a common tissue-engineering strategy for enhanced regeneration, the characteristics of the protein-biomaterial interface have not been fully elucidated. This study characterized the interface formed by the adsorbed proteins from various hemoderivatives with pristine and calcium phosphate (CaP)-coated polycaprolactone (PCL) melt electrowritten scaffolds. PCL scaffolds were fabricated by using melt electrospinning writing (MEW). Three hemoderivatives (pure platelet-rich plasma (P-PRP), leucocyte platelet-rich plasma (L-PRP) and injectable platelet-rich fibrin (i-PRF)) and total blood PLASMA (control) were prepared from ovine blood. Hemoderivatives were characterized via SEM/EDX, cross-linking assay, weight loss, pH and protein quantification. The interface between PCL/CaP and hemoderivative was examined via FTIR, XPS and electrophoresis. i-PRF/PCL-CaP (1653 cm-1), PLASMA/PCL-CaP (1652 cm-1) and i-PRF/PCL (1651 cm-1) demonstrated a strong signal at the Amide I region. PLASMA and i-PRF presented similar N1s spectra, with most of the nitrogen involved in N-C=O bonds (≈400 eV). i-PRF resulted in higher adsorption of low molecular weight (LMW) proteins at 60 min, while PLASMA exhibited the lowest adsorption. L-PRP and P-PRP had a similar pattern of protein adsorption. The characteristics of biomaterial interfaces can be customized, thus creating a specific hemoderivative-defined layer on the PCL surface. i-PRF demonstrated a predominant adsorption of LMW proteins. Further investigation of hemoderivative functionalized biomaterials is required to identify the differential protein corona composition, and the resultant immune response and regenerative capacity.

Fluid-assisted one-step fabrication of fused deposition molding 3D printing parts with conductive networks and gradient functionalities

The steps of preparing fused deposition molding (FDM) 3D printing conductive materials by melt-compounding are cumbersome, and the addition of fillers degrades the bulk properties of the materials, which limits the promotion and application of FDM 3D printing conductive materials. The FDM 3D printing conductive parts can be prepared by embedding conductive fillers on the surface of the printing filaments or parts, but available preparation methods are difficult to accomplish in one step. Herein, we introduce a facile method of one-step preparation of conductive parts by fluid-assisted FDM 3D printing. In this method, a thin tube transports conductive fluid, loads conductive carbon nanotubes (CNTs) onto the surface of molten polycaprolactone (PCL) filaments at the nozzle, and forms the CNT layers to prepare conductive parts in one step. The SEM and Raman results show that the CNTs are embedded on the surface of the PCL filaments. The sample can obtain a conductivity of 4.24 S m−1 loading only 4 mg CNTs. It also revealed that the preparation method has little effect on the mechanical properties of the FDM 3D printed PCL parts. The results exhibited that the as-proposed one-step preparation of conductive parts by fluid-assisted 3D printing could provide a new method for FDM 3D printing and fabricating high-performance parts, which has great potentials in various fields.

Osteo-Immunomodulatory Role of Interleukin-4-Immobilized Plasma Immersion Ion Implantation Membranes for Bone Regeneration.

Barrier membranes for guided tissue regeneration are essential for bone repair and regeneration. The implanted membranes may trigger early inflammatory responses as a foreign material, which can affect the recruitment and differentiation of bone cells during tissue regeneration. The purpose of this study was to determine whether immobilizing interleukin 4 (IL4) on plasma immersion ion implantation (PIII)-activated surfaces may alter the osteo-immunoregulatory characteristics of the membranes and produce pro-osteogenic effects. In order to immobilize IL4, polycaprolactone surfaces were modified using the PIII technology. No discernible alterations were found between the morphology before and after PIII treatment or IL4 immobilization. IL4-immobilized PIII surfaces polarized macrophages to an M2 phenotype and mitigated inflammatory cytokine production under lipopolysaccharide stimulation. Interestingly, the co-culture of macrophages (on IL4-immobilized PIII surfaces) and bone marrow-derived mesenchymal stromal cells enhanced the production of angiogenic and osteogenic factors and triggered autophagy activation. Exosomes produced by PIII + IL4-stimulated macrophages were also found to play a role in osteoblast differentiation. In conclusion, the osteo-immunoregulatory properties of bone materials can be modified by PIII-assisted IL4 immobilization, creating a favorable osteoimmune milieu for bone regeneration.

Modulating Myofibroblastic Differentiation of Fibroblasts through Actin-MRTF Signaling Axis by Micropatterned Surfaces for Suppressed Implant-Induced Fibrosis.

Myofibroblasts, the primary effector cells for implant-induced fibrosis, contribute to this process by secreting excessive collagen-rich matrix and contracting. Thus, approaches that suppress myofibroblasts may achieve desirable suppression effects in the fibrotic process. As one of the important physical properties of materials, material topographical structures have been proven to affect various aspects of cell behaviors, so is it possible to manipulate the formation of myofibroblasts by tailoring the topographical properties of medical devices? In this study, polycaprolactone (PCL) surfaces with typical micropatterns (micro column and micro pit) were fabricated. The regulatory effects of surface micropatterns on the myofibroblastic differentiation of fibroblasts were investigated. Compared to the flat surfaces and surfaces with micro pit, surfaces with micro columns triggered the F- to G-actin transition, inhibiting the nuclear transfer of myocardin-related transcription factor-A. Subsequently, the downstream gene α-smooth muscle actin, which is a marker of myofibroblasts, was suppressed. Further in vivo investigation showed that PCL implants with micro-column-patterned surfaces inhibited the formation of peri-implant fibrotic capsules. Our results demonstrate that surface topographical properties are a potent regulator of fibroblast differentiation into myofibroblasts and highlight the antifibrotic potential of modifying surfaces with micro-column patterns.

Biodegradable Mulching Films Based on Polycaprolactone and Its Porous Structure Construction.

Polycaprolactone (PCL) is one of the promising linear aliphatic polyesters which can be used as mulching film. Although it has suitable glass transition temperature and good biodegradability, further practical applications are restricted by the limited temperature-increasing and moisturizing properties. The rational design of the PCL structure is a good strategy to enhance the related properties. In this study, thermally-induced phase separation (TIPS) was introduced to fabricate a PCL nanoporous thin film. The introduction of a nanoporous structure on the PCL surface (np-PCL) exhibited enhanced temperature-increasing and moisturizing properties when used as mulch film. In detail, the average soil temperature of np-PCL was increased to 17.81 °C, when compared with common PCL of 17.42 °C and PBAT of 17.50 °C, and approaches to PE of 18.02 °C. In terms of water vapor transmission rate, the value for np-PCL is 637 gm-2day-1, which was much less than the common PCL of 786 and PBAT of 890 gm-2day-1. As a result, the weed biomass under the np-PCL was suppressed to be 0.35 kg m-2, almost half of the common PCL and PBAT. In addition, the np-PCL shows good thermal stability with an onset decomposition temperature of 295 °C. The degradation mechanism and rate of the np-PCL in different pH environments were also studied to explore the influence of nanoporous structure. This work highlights the importance of the nanoporous structure in PCL to enhance the temperature-increasing and moisturizing properties of PCL-based biodegradable mulching film.

EMSCs‐Seeded Micro‐Stripe Patterned Polycaprolactone Promoting Sciatic Nerve Regeneration

AbstractTransplantation of a synthetic polymer nerve conduit carrying stem cells is promising for treating peripheral nerve injury. In this study, a micropatterned inner surface of polycaprolactone (PCL) nerve conduit cellularized with nasal respiratory mucosa‐derived ectoderm mesenchymal stem cells (EMSCs) is adopted for peripheral nerve regeneration. The PCL microstructure grating is fabricated by soft nanoimprint lithography (NIL), whose templates can be easily transferred from the Ebeam photoresist pattern. The most suitable microstructure is carefully chosen to support EMSCs alignment and nerve regeneration. In vitro, the micropatterned PCL nerve conduit greatly enhanced the directional alignment of EMSCs and the MBP and S100 expression of myelin proteins. Additionally, the micropatterned conduit preseeded with EMSCs significantly stimulated the axon‐directed extension of neural stem cells. In vivo, micropatterned PCL conduits cotransplanted with EMSCs enhanced motor function and promoted sciatic nerve regeneration and extension in a rat model of sciatic nerve defect. Taken together, this “all‐in‐one” nerve guidance conduit makes sciatic nerve regeneration and morphological repairing accelerated, which has a potential clinical application in neural tissue engineering.

Effect of microarchitectural surface of polycaprolactone modified by reactive ion etching on osteogenic differentiation

The surface topology of a bioimplantable material is considered to be the most important influencing factor on the cytocompatibility and function. The aim of this study was to compare the cytocompatibility and osteogenic differentiation of polycaprolactone (PCL) surfaces with spiky and hill-and-valley (H-a-V) topologies fabricated by the Plasma reactive ion etching (RIE) technique. The RIE process was performed at a plasma power of 100 W and pressure of 100 mTorr using an oxygen flow rate of 100 sccm for 3 min (for Spiky) and 5 min (for H-a-V). The surface roughness and wettability of PCL were improved with the increase in the RIE treatment time. The focal cell adhesion, migration, and proliferation were increased in the H-a-V group compared to the Spiky group. The cell differentiation values (alkaline phosphatase (ALP) and Alizarin-red staining) were statistically significantly improved in H-a-V compared to those of Spiky. A western blot analysis demonstrated that H-a-V promoted cell differentiation compared to Spiky. These results suggest that the H-a-V topology is more favorable for cytocompatibility and osteogenic differentiation over the Spiky topology.

Effect of Functionalization of the Polycaprolactone Film Surface on the Mechanical and Biological Properties of the Film Itself.

The lack of suitable functional groups for cell adhesion on the surface of Polycaprolactone (PCL) is one of the main limitations in order to use PCL for biomedical applications. The aim of this research is to modify the PCL film surface using arginine, via an aminolysis reaction. In this regard, after PCL films formation by casting method, they were immersed in arginine solutions of various concentration at room temperature or then heated to 40 °C and in the presence of isopropanol or without it. To assess the structure of the modified surface, its wettability, and mechanical properties, methods of measuring the contact angle and the strip tensile test were used, and to compare the degree of attachment and the rate of cell proliferation, the method of fluorescent staining of cultured cells was used. The change in protein synthesis by cells on the modified surface was assessed using Western blotting. The results obtained show that the treatment of PCL films with an aqueous solution of arginine at room temperature for 1 day increases the hydrophilicity of the surface. Wherein surface modification led to a two-fold decrease of mechanical strength and flow stress, but elongation increase by about 30% for PCL films after modification in 0.5 M aqueous arginine solution at room temperature. Moreover, cell attachment and proliferation, as well as collagen synthesis, were significantly enhanced after arginine modification. The proposed simple and effective method for modifying PCL films with arginine significantly expands the possibilities for developing biocompatible scaffolds for tissue engineering.

Degradation of Polylactide and Polycaprolactone as a Result of Biofilm Formation Assessed under Experimental Conditions Simulating the Oral Cavity Environment.

Polylactide (PLA) and polycaprolactone (PCL) are biodegradable and bioabsorbable thermoplastic polymers considered as promising materials for oral applications. However, any abiotic surface used, especially in areas naturally colonized by microorganisms, provides a favorable interface for microbial growth and biofilm development. In this study, we investigated the biofilm formation of C. krusei and S. mutans on the surface of PLA and PCL immersed in the artificial saliva. Using microscopic (AFM, CLSM) observations and spectrometric measurements, we assessed the mass and topography of biofilm that developed on PLA and PCL surfaces. Incubated up to 56 days in specially prepared saliva and microorganisms medium, solid polymer samples were examined for surface properties (wettability, roughness, elastic modulus of the surface layer), structure (molecular weight, crystallinity), and mechanical properties (hardness, tensile strength). It has been shown that biofilm, especially S. mutans, promotes polymer degradation. Our findings indicate the need for additional antimicrobial strategies for the effective oral applications of PLA and PCL.

Fabrication and investigating in vivo wound healing property of coconut oil loaded nanofiber/hydrogel hybrid scaffold

Obtaining a sustainable drug delivery system is a challenging issue in biomedical science. This became even more important in the wound regeneration process due to its long treatment process. In this study, the calcium alginate (CaAlg) hydrogel is coated on the surface of polycaprolactone (PCL)/gelatin (Gel) nanofibers containing coconut oil (CO) using the impregnation method. The physical, chemical, and morphological properties of produced samples are investigated using different characterization techniques to verify the influence of hydrogel. Water contact angle, swelling ratio, and water vapor permeability measurements are used to evaluate the effect of hydrogel on the hydrophilicity of the proposed system. The cell viability test showed that the nanocomposite hydrogel is biocompatible and could improve wound healing. According to drug release studies, hydrogel addition to the nanofiber system plays an essential role in controlling CO release rate in the first 250 h. In vivo studies also indicated faster skin regeneration.

Characterization of AnCUT3, a plastic-degrading paucimannose cutinase from Aspergillus niger expressed in Pichia pastoris

Cutinases are hydrolytic enzymes secreted by phytopathogens to degrade cutin, the main polymeric component of plant cuticles. The multifaceted functionality of cutinases has allowed for their exploitation for catalytic reactions beyond their natural purpose. To diversify and expand the cutinase enzyme class, we identified five cutinase homologs from the saprotroph Aspergillus niger. One of these cutinases, AnCUT3, was over-expressed in Pichia pastoris and its biophysicochemical properties characterized. The purified recombinant AnCUT3 possessed an optimum temperature of 25 °C, an optimum pH of 5, and was stable at temperatures up to 50 °C (1 h incubation, melting point of 45.6 °C) and in a wide pH range. Kinetic studies of AnCUT3 using pNP ester substrates showed the highest catalytic efficiency, kcat/Km of 859 mM−1 s−1 toward p-nitrophenyl decanoate (C10). Although its calculated molecular mass is 27 kDa, AnCUT3 was expressed as two glycosylated proteins of molecular weights 24 and 50 kDa. Glycan profiling detected the presence of atypical paucimannose N-glycans (≤Man1-5GlcNAc) from recombinant AnCUT3, suggesting protein-dependent glycan processing of AnCUT3 in P. pastoris. AnCUT3 was also able to degrade and modify the surface of polycaprolactone and polyethylene terephthalate. Taken together, these features poise AnCUT3 as a potential biocatalyst for industrial applications.

Membrane supported poly(butylene adipate-co-terephthalate) nanofibrous matrices as cardiac patch: Effect of basement membrane for the fiber deposition and cellular behavior

Electrospun nanofibrous matrices are convenient biomaterials that mimic extracellular matrices for adhesion, migration, proliferation, and differentiation of cells. The aim of this study is to optimize the electrospinning parameters for poly(butylene adipate-co-terephthalate) (PBAT) nanofiber production by the response surface methodology (RSM) and to develop a suitable material design for the usability of PBAT in cardiac tissue engineering. Therefore, electrospun PBAT nanofibrous matrices collected on solvent-casted polycaprolactone (PCL) or PBAT basement membranes at optimized conditions. The attachment and proliferation behavior of the H9C2 rat cardiomyoblasts investigated on different PBAT and PCL surface features as a model cell line. For this purpose, neat PBAT and PCL films have been used comparatively with both random (R-PBAT) and aligned PBAT (A-PBAT) nanofibers coated films. The effect of polymer concentration, flow rate, applied voltage, and needle tip-connector distance on fiber diameter and alignment was examined in the electrospinning process and optimum processing parameters were determined by RSM. The PBAT nanofibers were spun on basement membranes with 10% (w/v) polymer concentration, 1 mL/h volumetric flow rate, 2000 rpm collector rotation velocity (for aligned ones), 15 kV applied voltage, and 20 cm needle tip-collector distance. The average diameter of random (R-PBAT) and aligned (A-PBAT) nanofibers was calculated as 555 ± 126 nm and 417 ± 137 nm. The mechanical test results showed that the alignment topography increased the elastic modulus of PBAT nanofibers compared to random matrices. The alignment of fibers found as 91% and 75% within the ± 10° range for A-PBAT/SC-PBAT and A-PBAT/SC-PCL, respectively. These findings showed that usage of PCL, instead of PBAT, as basement membrane decrease the alignment of deposited nanofibers. A 7-day cell culture study conducted with H9C2 cells seeded samples to investigate the influence of these differences on cell behavior. The results indicated that the alignment of fibers provides a suitable topography to proliferate and spread in myocyte morphology for H9C2 cells especially compared to neat films. Cellular behavior and nanofiber deposition have been affected by the usage of various basement membrane polymers. These findings demonstrated that the usage of basement membrane as support material provides the required thickness and mechanical properties to the aligned PBAT nanofiber matrices, and this double layer structure might be a promising candidate for cardiac tissue engineering with further studies.

The Impact of Graphene Oxide on Polycaprolactone PCL Surfaces: Antimicrobial Activity and Osteogenic Differentiation of Mesenchymal Stem Cell

In dentistry, bone regeneration requires osteoinductive biomaterial with antibacterial properties. Polycaprolactone (PCL) may be combined with different nanofillers including reduced graphene oxide (rGO). Here, the amount of rGO filler was defined to obtain a biocompatible and antibacterial PCL-based surface supporting the adhesion and differentiation of human mesenchymal stem cells (MSCs). Compounds carrying three different percentages of rGO were tested. Among all, the 5% rGO-PCL compound is the most bacteriostatic against Gram-positive bacteria. All scaffolds are biocompatible. MSCs adhere and proliferate on all scaffolds; however, 5% rGO-PCL surface supports the growth of cells and implements the expression of extracellular matrix components necessary to anchor the cells to the surface itself. Moreover, the 5% rGO-PCL surface has superior osteoinductive properties confirmed by the improved alkaline phosphatase activity, mineral matrix deposition, and osteogenic markers expression. These results suggest that 5% rGO-PCL has useful properties for bone tissue engineering purposes.