Contact Guidance Effect Research Articles

Construction of a Curcumin‐Loaded PLLA/PCL Micro‐Nano Conjugated Fibrous Membrane to Synergistically Prevent Postoperative Adhesion From Multiple Perspectives

AbstractPostoperative adhesion (POA) has emerged as a prevalent clinical challenge in soft tissue repair, emphasizing the critical need for preventive measures. However, the complex POA development process makes POA prevention from a single aspect insufficient. Hence, a curcumin‐loaded poly‐L‐lactic acid‐poly (caprolactone) micro‐nano conjugated fibrous membrane (PAPC MCFM (cur)) is engineered to synergistically prevent POA from multiple perspectives, in which poly (caprolactone) (PCL) nanofibers (118 ± 12 nm) with low orientation traverse the oriented poly‐L‐lactic acid (PLLA) microfibers (2.0 ± 0.3 µm). The PAPC MCFM not only significantly improves the mechanical properties of the anisotropic fibrous membrane (AIFM) that the modulus of elasticity and the tensile strength in the direction vertical to microfiber orientation increase by 4.5 and 13.0 times, respectively, but also can further enhance the “contact guidance effect” of AIFM, i.e., hindering fibroblast adhesion, proliferation, and differentiation to myofibroblast through inhibiting integrin β1 activation, vinculin expression and focal adhesion (FA) formation, and the nuclear localization activation of yes‐associated protein (YAP). Except for these effects, PAPC MCFM loading with 2.5 mg mL−1 curcumin can further prevent POA by delivering anti‐inflammatory, antioxidant, and antibacterial properties, and by suppressing fibrosis through decreased transforming growth factor‐β1(TGF‐β1) expression, showing effective POA prevention in rat abdominal cavity and rabbit dura mater models.

The promotion of vascular reconstruction by hierarchical structures in biodegradable small-diameter vascular scaffolds

Tissue engineering of small-diameter vessels remains challenging due to the inadequate ability to promote endothelialization and infiltration of smooth muscle cells (SMCs). Ideal vascular graft is expected to provide the ability to support endothelial monolayer formation and SMCs infiltration. To achieve this, vascular scaffolds with both orientation and dimension hierarchies were prepared, including hierarchically random vascular scaffold (RVS) and aligned vascular scaffold (AVS), by utilizing degradable poly(ε-caprolactone)-co-poly(ethylene glycol) (PCE) and the blend of PCE/gelatin (PCEG) as raw materials. In addition to the orientation hierarchy, dimension hierarchy with small pores in the inner layer and large pores in the outer layer was also constructed in both RVS and AVS to further investigate the promotion of vascular reconstruction by hierarchical structures in vascular scaffolds. The results show that the AVS with an orientation hierarchy that consists with the natural vascular structure had better mechanical properties and promotion effect on the proliferation of vascular cells than RVS, and also exhibited excellent contact guidance effects on cells. While the dimension hierarchy in both RVS and AVS was favorable to the rapid infiltration of SMCs in a short culture time in vitro. Besides, the results of subcutaneous implantation further demonstrate that AVS achieved a fully infiltrated outer layer with wavy elastic fibers-mimic strips formation by day 14, ascribing to hierarchies of aligned orientation and porous dimension. The results further indicate that the scaffolds with both orientation and dimension hierarchical structures have great potential in the application of promoting the vascular reconstruction.

GelMA micropattern enhances cardiomyocyte organization, maturation, and contraction via contact guidance.

Cardiac tissue engineering has emerged as a promising approach for restoring the functionality of damaged cardiac tissues following myocardial infarction. To effectively replicate the native anisotropic structure of cardiac tissues in vitro, this study focused on the fabrication of micropatterned gelatin methacryloyl hydrogels with varying geometric parameters. These substrates were evaluated for their ability to guide induced pluripotent stem cell-derived cardiomyocytes (CMs). The findings demonstrate that the mechanical properties of this hydrogel closely resemble those of native cardiac tissues, and it exhibits high fidelity in micropattern fabrication. Micropatterned hydrogel substrates lead to enhanced organization, maturation, and contraction of CMs. A microgroove with 20-μm-width and 20-μm-spacing was identified as the optimal configuration for maximizing the contact guidance effect, supported by analyses of nuclear orientation and F-actin organization. Furthermore, this specific micropattern design was found to promote CMs' maturation, as evidenced by increased expression of connexin 43 and vinculin, along with extended sarcomere length. It also enhanced CMs' contraction, resulting in larger contractile amplitudes and greater contractile motion anisotropy. In conclusion, these results underscore the significant benefits of optimizing micropatterned gelatin methacryloyl for improving CMs' organization, maturation, and contraction. This valuable insight paves the way for the development of highly organized and functionally mature cardiac tissues in vitro.

Laser surface texturing of Ti-cp and Ti6Al4V alloy for the improvement of fibroblast adhesion and alignment and the reduction of bacterial adhesion

Surface texturing is widely investigated for the modulation of the surface properties and biological response of implantable materials. Laser is a green technology for the obtainment of specific patterns without the employment of toxic chemicals. In the present research, a picosecond laser treatment was applied to commercially pure titanium and Ti6Al4V alloy for the obtainment of aligned grooves characterized by micrometric width and spacing but submicrometric depth to promote fibroblast alignment and discourage bacterial adhesion. Surface topography and roughness of laser-textured surfaces were characterized using confocal microscopy, contact profiler, and Scanning Electron Microscopy. Zeta potential titration curves and contact angle measurements were used for the investigation of surface charge and wettability, while X-Ray Diffraction was employed for crystallographic characterization. Finally, the biological response (fibroblast adhesion and alignment and bacterial adhesion) was assessed. Similar results were obtained on commercially pure Ti and Ti6Al4V. Grooves with the designed features were obtained (about 15 μm spaced and 0.2μ deep) and the treated surfaces had a complex micro- and nano-scale morphology which reduced wettability. No evident chemical modification occurred. The treated surfaces were biocompatible and grooves had a contact guidance effect on human fibroblasts. The obtained topography had anti-adhesive action on bacteria (Staphylococcus aureus).

Accelerated epithelial layer healing induced by tactile anisotropy in surface topography

Mammalian cells respond to tactile cues from topographic elements presented by the substrate. Among these, anisotropic features distributed in an ordered manner give directionality. In the extracellular matrix, this ordering is embedded in a noisy environment altering the contact guidance effect. To date, it is unclear how cells respond to topographical signals in a noisy environment. Here, using rationally designed substrates, we report morphotaxis, a guidance mechanism enabling fibroblasts and epithelial cells to move along gradients of topographic order distortion. Isolated cells and cell ensembles perform morphotaxis in response to gradients of different strength and directionality, with mature epithelia integrating variations of topographic order over hundreds of micrometers. The level of topographic order controls cell cycle progression, locally delaying or promoting cell proliferation. In mature epithelia, the combination of morphotaxis and noise-dependent distributed proliferation provides a strategy to enhance wound healing as confirmed by a mathematical model capturing key elements of the process.

Photofunctionalized TiO2-TiN micropattern coating with anticoagulant properties and ECs contact-guidance effect

Titanium nitride (TiN) and titanium dioxide (TiO2) are two titanium-based coatings commonly used in cardiovascular stent surface engineering. Generally, TiN has good mechanical properties and endothelial cell (ECs) compatibility but poor anticoagulant properties and cannot modulate cell growth orientation and morphology. TiO2 has excellent corrosion resistance and biosafety. Besides, TiO2 has the photocatalytic anticoagulant property, which can migrate to other materials tens of microns away. Based on the above properties, a striped TiO2-TiN micropattern coating was designed and fabricated in this study, and the coating was photofunctionalized by UV irradiation. The obtained photo-functionalized TiO2-TiN micropattern coating showed anticoagulant properties by the migrating effect of the photocatalytic anticoagulant property of TiO2. Besides, the TiO2-TiN micropattern coatings showed ECs compatibility. Furthermore, the growth orientation and cell shape of ECs on TiO2-TiN samples were effectively regulated by the stripe pattern's contact guidance effect, which was particularly evident on the photo-functionalized TiO2-TiN samples. We envision that this photofunctionalized TiO2-TiN striped micropattern coating has significant potential for the surface engineering of vascular stents.

Contact Guidance Effect and Prevention of Microfouling on a Beta Titanium Alloy Surface Structured by Electron-Beam Technology.

Nano- and micro-structuring of implantable materials constitute a promising approach to introduce mechanical contact guidance effect, drive cells colonization, as well as to prevent bacteria adhesion and biofilm aggregation, through antifouling topography. Accordingly, this paper aims to extend the application of e-beam surface texturing and nano-structuring to the beta titanium alloys, which are of great interest for biomedical implants because of the low Young modulus and the reduction of the stress shielding effect. The paper shows that surface texturing on the micro-scale (micro-grooves) is functional to a contact guidance effect on gingival fibroblasts. Moreover, nano-structuring, derived from the e-beam surface treatment, is effective to prevent microfouling. In fact, human fibroblasts were cultivated directly onto grooved specimens showing to sense the surface micro-structure thus spreading following the grooves’ orientation. Moreover, Staphylococcus aureus colonies adhesion was prevented by the nano-topographies in comparison to the mirror-polished control, thus demonstrating promising antifouling properties. Furthermore, the research goes into detail to understand the mechanism of microfouling prevention due to nano-topography and microstructure.

Cell-friendly photo-functionalized TiO2 nano-micro-honeycombs for selectively preventing bacteria and platelet adhesion

Titanium dioxide (TiO2) is a widely used biomaterial. It is a great challenge to confer antibacterial and antithrombotic properties to TiO2 while maintaining its cell affinity. Here, we developed a new strategy to achieve the above goal by comprehensively controlling the chemical cues and geometrical cues of the surface of TiO2. Using colloidal etching technology and UV irradiation treatment, we obtained the photofunctionalized nano-micro-honeycomb structured TiO2. The honeycomb structured increased the photocatalytic activity of TiO2, which endowed TiO2 with photo-induced superhydrophilicity to inhibit bacterial adhesion. The high photocatalytic activity also induced the strong photocatalytic oxidation of TiO2 surface organic adsorbates to suppress fibrinogen and platelet attachment. In addition, owing to the micropore trapping-isolation effect on the bacteria and the nano-frames' contact guidance effect on the growth and spreading of platelet pseudopods, the honeycomb structure also shows a considerable inhibiting effect on bacterial and platelet adhesion. Therefore, due to the controlled chemical and geometrical cues' synergistic effect, the photo-functionalized TiO2 honeycomb structure shows excellent bacterial-adhesion resistance and antithrombotic properties. More importantly, the photo-functionalized TiO2 honeycomb did not inhibit the adhesion and growth of endothelial cells (ECs) after culturing for 3 d, indicating a good cell affinity that the traditional antifouling surfaces do not possess.

Stiffness of the aligned fibers affects structural and functional integrity of the oriented endothelial cells

Promoting healthy endothelialization of the tissue-engineered vascular grafts is of great importance in preventing the occurrence of undesired post-implantation complications including neointimal hyperplasia, late thrombosis, and neoatherosclerosis. Previous researches have demonstrated the crucial role of scaffold topography or stiffness in modulating the behavior of the monolayer endothelial cells (ECs). However, effects of the stiffness of scaffolds with anisotropic topography on ECs within vivo like oriented morphology has received little attention. In this study, aligned fibrous substrates (AFSs) with tunable stiffness (14.68–2141.72 MPa), similar to the range of stiffness of the healthy and diseased subendothelial matrix, were used to investigate the effects of fiber stiffness on ECs’ attachment, orientation, proliferation, function, remodeling and dysfunction. The results demonstrate that stiffness of the AFSs, capable of providing topographical cues, is a crucial endothelium-protective microenvironmental factor by maintaining stable and quiescent endothelium with in vivo like orientation and strong cell–cell junctions. Stiffer AFSs exacerbated the disruption of endothelium integrity, the occurrence of endothelial-to-mesenchymal transition (EndMT), and the inflammation-induced activation in the endothelial monolayer. This study provides new insights into the understanding on how the stiffness of biomimicking anisotropic substrate regulates the structural and functional integrity of the in vivo like endothelial monolayer, and offers essential designing parameters in engineering biomimicking small-diameter vascular grafts for the regeneration of viable blood vessels. Statement of significanceIn vascular tissue engineering, promoting endothelialization on scaffold surface has been considered as a paramount strategy to reduce post-implantation complications. Electrospun aligned fibers have been known to provide contact guidance effect in directing endothelial cells' oriented growth, however, whether the formed EC monolayer in ‘correct’ orientation shape is of ‘correct’ function hasn't been explored yet. Given the recognized important role of substrate stiffness in endothelial function, AFSs across physiologically relevant range of moduli (14.68–2141.72 MPa) while maintaining consistent surface chemistry and topographical features were employed to investigate the fiber stiffness effects on ECs function in anisotropic morphology. This study will provide more insightful perspectives in the physiologically remodeling progression of vascular endothelium and design of vascular scaffolds.

Effects of aligned electrospun fibers with different diameters on hemocompatibility, cell behaviors and inflammation in vitro

An endothelial cell (EC) monolayer aligned along the direction of blood flow in vivo shows excellent capacity for anti-inflammation and anti-thrombosis. Therefore, aligned electrospun fibers have been much studied in the field of vascular implants since they are considered to facilitate the formation of an aligned EC monolayer, yet few research studies have been comprehensively reported concerning the effects of diameter scales of aligned fibers. In the present work, a series of aligned polycaprolactone (PCL) electrospun fibers with varying diameters ranging from dozens of nanometers to several micrometers were developed, and the effects of the fiber scales on EC behaviors, hemocompatibility as well as inflammatory cell behaviors were investigated, to evaluate their potential performance in the field of vascular implants. Our results showed that platelets exhibited small attachment forces on all fibers, and the anticoagulation property improved with the decrease of the fiber diameters. The impact of fiber diameters on human umbilical vein endothelial cell (HUVEC) adhesion and NO release was limited, while significant on HUVEC proliferation. With the increase of the fiber diameters, the elongation of HUVECs on our samples increased first then decreased, and exhibited maximum elongation degrees on 2738 nm and 2036 nm due to the strong contact guidance effect on these graphical cues; too thick or too fine fibers would weaken the contact guidance effect. Furthermore, we hypothesized that HUVECs cultured on 2036 nm had the smallest spreading area because of their elongation, but 2738 nm restricted HUVECs spreading limitedly. Similarly, NO production of HUVECs showed a similar change trend as their elongation degrees on different fibers. Except for 2036 nm, it exhibited the second highest NO production. For RAW 264.7 cells, poorer cell adhesion and lower TNF-α concentration of 1456 nm indicated its superior anti-inflammation property, while 73 nm showed a contrasting performance. Overall, these findings partly revealed the relationship between different topographies and cell behaviors, providing basic insight into vascular implant design.

Engineering High-Resolution Micropatterns Directly onto Titanium with Optimized Contact Guidance to Promote Osteogenic Differentiation and Bone Regeneration.

Topographical cues play an important role in directing cell behavior, and thus, extensive research efforts have been devoted to fabrication of surface patterns and exploring the contact guidance effect. However, engineering high-resolution micropatterns directly onto metallic implants remains a grand challenge. Moreover, there still lacks evidence that allows translation of in vitro screening to in vivo tissue response. Herein, we demonstrate a fast, cost-effective, and feasible approach to the precise fabrication of shape- and size-controlled micropatterns on titanium substrates using a combination of photolithography and inductively coupled plasma-based dry etching. A titanium TopoChip containing 34 microgrooved patterns with varying geometry parameters and a flat surface as the control was designed for a high-throughput in vitro study of the contact guidance of osteoblasts. The correlation between the surface pattern dimensions, cell morphological characteristics, proliferation, and osteogenic marker expression was systematically investigated in vitro. Furthermore, the surface with the highest osteogenic potential in vitro along with representative controls was evaluated in rat cranial defect models. The results show that microgrooved pattern parameters have almost no effect on osteoblast proliferation but significantly regulate the cell morphology, orientation, focal adhesion (FA) formation, and osteogenic differentiation in vitro. In particular, a specific groove pattern with a ridge width of 3 μm, groove width of 7 μm, and depth of 2 μm can most effectively align the cells through regulating the distribution of FAs, resulting in an anisotropic actin cytoskeleton, and thereby promoting osteogenic differentiation. In vivo, microcomputed tomography and histological analyses show that the optimized pattern can apparently stimulate new bone formation. This study not only offers a microfabrication method that can be extended to fabricate various shape- and size-controlled micropatterns on titanium alloys but also provides insight into the surface structure design of orthopedic and dental implants for enhanced bone regeneration.

Modeling cell migration regulated by cell extracellular-matrix micromechanical coupling.

Cell migration in fibrous extracellular matrix (ECM) is crucial to many physiological and pathological processes such as tissue regeneration, immune response, and cancer progression. During migration, individual cells can generate active pulling forces via actomyosin contraction, which are transmitted to the ECM fibers through focal adhesion complexes, remodel the ECM, and eventually propagate to and can be sensed by other cells in the system. The microstructure and physical properties of the ECM can also significantly influence cell migration, e.g., via durotaxis and contact guidance. Here, we develop a computational model for two-dimensional cell migration regulated by cell-ECM micromechanical coupling. Our model explicitly takes into account a variety of cellular-level processes, including focal adhesion formation and disassembly, active traction force generation and cell locomotion due to actin filament contraction, transmission and propagation of tensile forces in the ECM, as well as the resulting ECM remodeling. We validate our model by accurately reproducing single-cell dynamics of MCF-10A breast cancer cells migrating on collagen gels and show that the durotaxis and contact guidance effects naturally arise as a consequence of the cell-ECM micromechanical interactions considered in the model. Moreover, our model predicts strongly correlated multicellular migration dynamics, which are resulted from the ECM-mediated mechanical coupling among the migrating cell and are subsequently verified in in vitro experiments using MCF-10A cells. Our computational model provides a robust tool to investigate emergent collective dynamics of multicellular systems in complex in vivo microenvironment and can be utilized to design in vitro microenvironments to guide collective behaviors and self-organization of cells.

Surface structuring by Electron Beam for improved soft tissues adhesion and reduced bacterial contamination on Ti-grade 2

Even if soft tissue adhesion on titanium surfaces is an issue of great interest for transmucosal dental implants and percutaneous devices, few specific surface modification strategies have been approached so far to face it. Topographical surface modification is an interesting strategy because quality of soft tissues can be effectively improved by contact guidance effect on fibroblasts. Conversely, biofilm formation is encouraged in presence of rough or porous surfaces, as well as of surface patterns, if the roughness overcome the threshold of 0.2 μm. This issue becomes particularly critical in medical devices which cross different biological tissues and, through soft tissues, establish a direct contact with the external environment. In the present research, three different grooved patterns (5, 10 and 30 μm wide) were produced on commercially pure titanium by means of Electron Beam (EB) structuring maintaining the average surface roughness lower than 0.2 μm. The EB surface structured samples were characterized by means of optical and atomic force microscopy, contact and non-contact profilometry, X-Ray Diffraction, residual stress measurements as well as bacteria and cell adhesion tests. The 10 μm wide grooves resulted able to support fibroblast alignment onto the metallic substrate. Unexpectedly, all the EB surface structured surfaces not only did not enhance, but even reduced bacterial adhesion up to 48 h culture.

Mesoscale substrate curvature overrules nanoscale contact guidance to direct bone marrow stromal cell migration.

The intrinsic architecture of biological tissues and of implanted biomaterials provides cells with large-scale geometrical cues. To understand how cells are able to sense and respond to complex structural environments, a deeper insight into the cellular response to multi-scale and conflicting geometrical cues is needed. In this study, we subjected human bone marrow stromal cells (hBMSCs) to mesoscale cylindrical surfaces (diameter 250-5000 µm) and nanoscale collagen fibrils (diameter 100-200 nm) that were aligned perpendicular to the cylinder axis. On flat surfaces and at low substrate curvatures (cylinder diameter d > 1000 µm), cell alignment and migration were governed by the nanoscale collagen fibrils, consistent with the contact guidance effect. With increasing surface curvature (decreasing cylinder diameter, d < 1000 µm), cells increasingly aligned and migrated along the cylinder axis, i.e. the direction of zero curvature. An increase in phosphorylated myosin light chain levels was observed with increasing substrate curvature, suggesting a link between substrate-induced cell bending and the F-actin-myosin machinery. Taken together, this work demonstrates that geometrical cues of up to 10× cell size can play a dominant role in directing hBMSC alignment and migration and that the effect of nanoscale contact guidance can even be overruled by mesoscale curvature guidance.

Effects of aligned and random fibers with different diameter on cell behaviors.

Tissue-engineered vascular grafts show promise as a treatment for patients with diseased or damaged arteries. The formation of a confluent endothelium lumen on scaffold is one of the key issues in vascular tissue engineering. The endothelial layer is composed of endothelial cells (ECs) aligned along the long axis of the vessel. The contact guidance effect is believed to fundamentally impact cell morphology, alignment, and behavior. By tuning the scaffold topography where the EC cells reside, the cell adhesion, shape, and other behaviors might be regulated. To this end, poly(ε-caprolactone) was electrospun to obtain different topographies by varying fiber diameter and orientation. The morphology and alignment of EC cells were investigated by analyzing fluorescence staining images. The results show that nano-sized fibers significantly enhance cell adhesion and proliferation due to the comparable geometric feature size of the fiber diameter. ECs align along the fiber oriented direction for both microfibers and nanofibers. It shows a better alignment of ECs on micro-aligned fibers than on nano-aligned fibers, which is due to the active size range of the contact guidance effect. These results indicate that scaffolds with ordered topographies might favor the formation of a confluent endothelium for vascular tissue engineering.

Cell-instructive high-resolution micropatterned polylactic acid surfaces

Micro and nanoscale topographical structuring of biomaterial surfaces has been a valuable tool for influencing cell behavior, including cell attachment, proliferation and differentiation. However, most fabrication techniques for surface patterning of implantable biomaterials suffer from a limited resolution, not allowing controlled generation of sub-cellular three-dimensional features. Here, a direct laser lithography technique based on two-photon absorption was used to construct several patterns varying in size between 500 nm and 15 μm. Through replication via an intermediate mold, the patterns were transferred into polylactic acid (PLA), a widely used biomedical polymer, while retaining the original geometry. An osteoblast-like cell line, MG-63 was used for characterizing the morphological response to the topographical patterns. The results indicated that semi-continuous (dashed) lines, with a height of 1 μm were able to induce cell elongation in the direction of the lines. However, when dashes with a height of 0.5 μm were combined with perpendicularly crossing continuous lines (rails) with a height of 8 μm, the contact guidance effect of the dashes was lost and elongation of the cells was observed in the direction of the larger features. A second pattern, consisting of different arrays of pillars showed that, depending on the pillar height, the cells were either able to spread over the pattern or were confined between the pattern features. These differences in the ability of cells to spread further resulted in the formation of tension forces through stress fibers and displacement of vimentin. The method for high-resolution micropatterning of PLA as presented here can also be applied to other biomedical polymers, making it useful both for fundamental studies and for designing new biomaterials with improved functionality.

Easy fabrication of aligned PLLA nanofibers-based 2D scaffolds suitable for cell contact guidance studies

An easy, low-cost and fast wet processing-based method named ASB-SANS (Auxiliary Solvent-Based Sublimation-Aided NanoStructuring) has been used to fabricate poly(l-lactic acid) (PLLA) highly ordered and hierarchically organized 2D fibrillar patterns, with fiber widths between 40 and 500nm and lengths exceeding tens of microns. A clear contact guidance effect of these nanofibrillar scaffolds with respect to HeLa and NIH-3T3 cells growth has been observed, on top of an overall good viability. For NIH-3T3 pronounced elongation of the cells was observed, as well as a remarkable ability of the patterns to guide the extension of pseudopodia. Moreover, SEM imaging revealed filopodia stemming from both sides of the pseudopodia and aligned with the secondary PLLA nanofibrous structures created by the ASB-SANS procedure. These results validate ASB-SANS as a technique capable to provide biocompatible 2D nanofibrillar patterns suitable for studying phenomena of contact guidance (and, more in general, the behavior of cells onto nanofibrous scaffolds), at very low costs and in an extremely easy way, accessible to virtually any laboratory.

Precise patterning of biopolymers and cells by direct write technique

It is demonstrated how micro-periodic patterns can be precisely created using biopolymer solutions of optimised rheological properties. Direct-write assembly enabled the construction of micro-periodic structures without the need for expensive tooling, dies, or lithographic masks. It is reported that rationalised designer patterns of defined filamentous topography could provide contact guidance effect for alignment of corneal stromal cells and dermal fibroblasts, and control cellular morphology to simulate organisation of cells and extracellular matrix in native tissues.

Hybrid Microfabrication of Nanofiber-Based Sheets and Rods for Tissue Engineering Applications

Electrospun nanofibers have been developed into a variety of forms for tissue engineering scaffolds to regulate the cellular functions guided by nanotopographical cues. Here, we have successfully fabricated nanofiber-based scaffold complexes of rod and sheet type by combining the three microfabrication techniques of electrospinning, spin coating, and polymer melt deposition. It was demonstrated that this hybrid fabrication could produce uniaxially aligned nanofiber scaffolds supported by a thin film, allowing for a mechanically enforced substrate for cell culture as well as facile scaffold manipulation. The results of cell analysis indicated that nanofibers on spin-coated films could provide contact guidance effects on cells and retain them even after manipulation. As an application of the cell-laden nanofiber film, we built a rod-type structure by rolling up the film around a mechanically supporting core microfiber, which was incorporated by polymer melt deposition. A biocompatible and biodegradable polymer, polycaprolactone, was used throughout the processes and thus could be used as a directly implantable substitute in tissue regeneration.

To enhance protein production from osteoblasts by using micro-patterned surfaces

In this study, microgrooved surfaces were used to culture osteoblast-like cells to promote the production of osteogenic proteins. First, modified photolithographic techniques were used on silicon wafers to create micro-grooved patterns. Groove width was continuously varied from 1 to 20μm. Then, the cells were cultured for various periods to induce the contact guidance effect, and the cell behaviors, including the production of osteogenic proteins, were analyzed. The results indicated that the contact guidance effect was significant with suitable groove width. The guidance from topography promoted cell attachment and proliferation. Moreover, the secretion of osteogenic proteins, including alkine phosphate (ALP) and osteopontin (OPN), was greatly enhanced after cells were guided by grooved patterns. The grooved structure induced osteocalcin gene expression, revealed by the promotion of phosphorylation of Smad1. It was found that groove width less than 10μm induced the alignment of osteoblasts, causing the increase in the production of osteogenic proteins.