Nanotopographical Features Research Articles

Biomechanics of Macrophages on Disordered Surface Nanotopography.

Macrophages are involved in every stage of the innate/inflammatory immune responses in the body tissues, including the resolution of the reaction, and they do so in close collaboration with the extracellular matrix (ECM). Simplified substrates with nanotopographical features attempt to mimic the structural properties of the ECM to clarify the functional features of the interaction of the ECM with macrophages. We still have a limited understanding of the macrophage behavior upon interaction with disordered nanotopography, especially with features smaller than 10 nm. Here, we combine atomic force microscopy (AFM), finite element modeling (FEM), and quantitative biochemical approaches in order to understand the mechanotransduction from the nanostructured surface into cellular responses. AFM experiments show a decrease of macrophage stiffness, measured with the Young's modulus, as a biomechanical response to a nanostructured (ns-) ZrOx surface. FEM experiments suggest that ZrOx surfaces with increasing roughness represent weaker mechanical boundary conditions. The mechanical cues from the substrate are transduced into the cell through the formation of integrin-regulated focal adhesions and cytoskeletal reorganization, which, in turn, modulate cell biomechanics by downregulating cell stiffness. Surface nanotopography and consequent biomechanical response impact the overall behavior of macrophages by increasing movement and phagocytic ability without significantly influencing their inflammatory behavior. Our study suggests a strong potential of surface nanotopography for the regulation of macrophage functions, which implies a prospective application relative to coating technology for biomedical devices.

Smart Biointerfaces via Click Chemistry-Enabled Nanopatterning of Multiple Bioligands and DNA Force Sensors.

Nanoscale biomolecular placement is crucial for advancing cellular signaling, sensor technology, and molecular interaction studies. Despite this, current methods fall short in enabling large-area nanopatterning of multiple biomolecules while minimizing nonspecific interactions. Using bioorthogonal tags at a submicron scale, we introduce a novel hole-mask colloidal lithography method for arranging up to three distinct proteins, DNA, or peptides on large, fully passivated surfaces. The surfaces are compatible with single-molecule fluorescence microscopy and microplate formats, facilitating versatile applications in cellular and single-molecule assays. We utilize fully passivated and transparent substrates devoid of metals and nanotopographical features to ensure accurate patterning and minimize nonspecific interactions. Surface patterning is achieved using bioorthogonal TCO-tetrazine (inverse electron-demand Diels-Alder, IEDDA) ligation, DBCO-azide (strain-promoted azide-alkyne cycloaddition, SPAAC) click chemistry, and biotin-avidin interactions. These are arranged on surfaces passivated with dense poly(ethylene glycol) PEG brushes crafted through the selective and stepwise removal of sacrificial metallic and polymeric layers, enabling the directed attachment of biospecific tags with nanometric precision. In a proof-of-concept experiment, DNA tension gauge tether (TGT) force sensors, conjugated to cRGD (arginylglycylaspartic acid) in nanoclusters, measured fibroblast integrin tension. This novel application enables the quantification of forces in the piconewton range, which is restricted within the nanopatterned clusters. A second demonstration of the platform to study integrin and epidermal growth factor (EGF) proximal signaling reveals clear mechanotransduction and changes in the cellular morphology. The findings illustrate the platform's potential as a powerful tool for probing complex biochemical pathways involving several molecules arranged with nanometer precision and cellular interactions at the nanoscale.

Biological Performance of Titanium Surfaces with Different Hydrophilic and Nanotopographical Features

Biological Performance of Titanium Surfaces with Different Hydrophilic and Nanotopographical Features

Oriented fibrous poly (butylene adipate-co-terephthalate) matrices with nanotopographic features: Production and characterization

This is the first study to systematically investigate the effectiveness of electrospinning parameters and solvent properties on the random and aligned poly (butylene adipate-co-terephthalate) (PBAT) fiber surface nanotopography. Therefore, the combinations of three groups of solvents, good solvents, and a non-solvent, were used. While THF (tetrahydrofuran), HFIP (1,1,1,3,3,3-hexafluoro-2-propanol) and DCM (dichloromethane) were selected as good solvents, DMSO (dimethylsulfoxide) was chosen as a non-solvent. Four different phase separation mechanisms, non-solvent-induced phase separation (NIPS), vapor-induced phase separation (VIPS), breath figure (BF), and thermally induced phase separation (TIPS) mechanisms, were applied to create roughness on the fiber surfaces. Smooth, porous and groove surfaces with average fiber diameters in the range of about 800–2000 nm were obtained at different electrospinning conditions by these phase separation mechanisms. Porous random fibers with a few small pores were obtained by NIPS in the presence of THF/DMSO, while corrugated random fibers were produced by VIPS with DCM/HFIP. The groove roughness on the aligned THF/DMSO and aligned DCM/HFIP fibers was achieved by the TIPS mechanism thanks to the rapid evaporation rate of THF and HFIP. The most porous fibers were obtained in the presence of moisture with a good solvent mixture of DCM/HFIP. The results of this investigation show that the use of good and non-solvents together with moisture control affected not only the topography of fibrous matrices but also the bulk characteristics of PBAT nanofibrous matrices.

Catalyst Droplet‐Based Puncturable Nanostructures with Mechano‐Bactericidal Properties Against Bioaerosols

AbstractBioaerosol contamination problems have led to the need for new technologies that effectively collect and inactivate airborne microorganisms. Typical nanomaterial‐based filter membranes are usually sterilized using photocatalysts, electrical stimulation, and thermal treatment, which are expensive and require additional devices and cumbersome manufacturing. In this study, a membrane with nanotopographical features is manufactured via a catalyst droplet‐based procedure to mechanically damage airborne bacteria. The catalyst droplets are used as templates for in situ novel puncturable nanopillar growth on the membrane surface. Numerical simulations and microscopic observations show that puncturable nanopillars with a thin and rough nano‐edge are advantageous for rupturing the bacterial cell compared to flat nanopillars without a thin edge. A puncturable nanostructured air filter (PNAF) is compared to a bare air filter and exhibits higher bioaerosol collection efficiencies (>98% and 89.3–95.7%, respectively). PNAF is tested under breathing conditions as part of a face mask, where it effectively captures and deactivates E. coli aerosols through a mechano‐bactericidal effect, resulting in the inhibition of bacterial proliferation and finally death. Thus, PNAF can be applied as an air purifier or face mask filter for bioaerosol collection presenting antibacterial effects without external stimulation.

Nanotopography and Microconfinement Impact on Primary Hippocampal Astrocyte Morphology, Cytoskeleton and Spontaneous Calcium Wave Signalling.

Astrocytes' organisation affects the functioning and the fine morphology of the brain, both in physiological and pathological contexts. Although many aspects of their role have been characterised, their complex functions remain, to a certain extent, unclear with respect to their contribution to brain cell communication. Here, we studied the effects of nanotopography and microconfinement on primary hippocampal rat astrocytes. For this purpose, we fabricated nanostructured zirconia surfaces as homogenous substrates and as micrometric patterns, the latter produced by a combination of an additive nanofabrication and micropatterning technique. These engineered substrates reproduce both nanotopographical features and microscale geometries that astrocytes encounter in their natural environment, such as basement membrane topography, as well as blood vessels and axonal fibre topology. The impact of restrictive adhesion manifests in the modulation of several cellular properties of single cells (morphological and actin cytoskeletal changes) and the network organisation and functioning. Calcium wave signalling was observed only in astrocytes grown in confined geometries, with an activity enhancement in cells forming elongated agglomerates with dimensions typical of blood vessels or axon fibres. Our results suggest that calcium oscillation and wave propagation are closely related to astrocytic morphology and actin cytoskeleton organisation.

Molecular mechanisms of interrod spacing-mediated osseointegration via modulating inflammatory response and osteogenic differentiation

Osseointegration is initiated by immune cell-derived inflammatory responses to the surfaces of implanted materials. However, the bio-molecular mechanisms by which nanotopographic features of implant surfaces immunomodulate osseointegration remain to be clarified. To this end, herein, three kinds of TiO2 nanorod-like arrays on Ti with the interrod spacing of ∼0, 45 and 85 nm, termed as S0, S45 and S85, respectively, were hydrothermally grown, and their actions on the polarization of macrophages (MΦs) and MΦs-mediated contact osteogenesis of bone marrow-derived mesenchymal stromal cells (BMSCs) were explored, especially the involved signaling pathways. Compared to pure Ti (namely P-Ti), the nanorods-arrayed Ti activated FG-integrin αMβ2-IKK-NF-κB signaling pathway in the adhered MΦs at initial 6 h while FN-integrin α5β1-PI3K-Akt1 pathway since 24 h, accelerating the transition of MΦs from pro-inflammatory (M1) to anti-inflammatory (M2) phenotype. With the cytokines secreted by M2 cells, the coatings activated the BMP2-TGFβ1-SMAD signaling pathway to enhance osteogenic differentiation of the recruited BMSCs in vitro. Also, the nanorods-coated Ti induced a proper immune microenvironment and accelerated bone apposition in rabbit tibia marrow cavities in vivo. Furthermore, owing to the favorable capacity of osteoimmunomodulation, the ∼85 nm spaced nanorods-arrayed coating exhibited satisfactorily in orchestrating osteogenic differentiation in vitro and in vivo. This work provides insights into nanotopographic features of implant surfaces immunomodulating contact osteogenesis.

The glycocalyx affects the mechanotransductive perception of the topographical microenvironment

The cell/microenvironment interface is the starting point of integrin-mediated mechanotransduction, but many details of mechanotransductive signal integration remain elusive due to the complexity of the involved (extra)cellular structures, such as the glycocalyx. We used nano-bio-interfaces reproducing the complex nanotopographical features of the extracellular matrix to analyse the glycocalyx impact on PC12 cell mechanosensing at the nanoscale (e.g., by force spectroscopy with functionalised probes). Our data demonstrates that the glycocalyx configuration affects spatio-temporal nanotopography-sensitive mechanotransductive events at the cell/microenvironment interface. Opposing effects of major glycocalyx removal were observed, when comparing flat and specific nanotopographical conditions. The excessive retrograde actin flow speed and force loading are strongly reduced on certain nanotopographies upon strong reduction of the native glycocalyx, while on the flat substrate we observe the opposite trend. Our results highlight the importance of the glycocalyx configuration in a molecular clutch force loading-dependent cellular mechanism for mechanosensing of microenvironmental nanotopographical features.Graphical

TiO2 Nanotopography-Driven Osteoblast Adhesion through Coulomb's Force Evolution.

Nanotopography is an effective method to regulate cells’ behaviors to improve Ti orthopaedic implants’ in vivo performance. However, the mechanism underlying cellular matrix–nanotopography interactions that allows the modulation of cell adhesion has remained elusive. In this study, we have developed novel nanotopographic features on Ti substrates and studied human osteoblast (HOb) adhesion on nanotopographies to reveal the interactive mechanism regulating cell adhesion and spreading. Through nanoflat, nanoconvex, and nanoconcave TiO2 nanotopographies, the evolution of Coulomb’s force between the extracellular matrix and nanotopographies has been estimated and comparatively analyzed, along with the assessment of cellular responses of HOb. We show that HObs exhibited greater adhesion and spreading on nanoconvex surfaces where they formed super matured focal adhesions and an ordered actin cytoskeleton. It also demonstrated that Coulomb’s force on nanoconvex features exhibits a more intense and concentrated evolution than that of nanoconcave features, which may result in a high dense distribution of fibronectin. Thus, this work is meaningful for novel Ti-based orthopaedic implants’ surface designs for enhancing their in vivo performance.

Nano-Structured Ridged Micro-Filaments (≥100 µm Diameter) Produced Using a Single Step Strategy for Improved Bone Cell Adhesion and Proliferation in Textile Scaffolds.

Textile scaffolds that are either 2D or 3D with tunable shapes and pore sizes can be made through textile processing (weaving, knitting, braiding, nonwovens) using microfilaments. However, these filaments lack nano-topographical features to improve bone cell adhesion and proliferation. Moreover, the diameter of such filaments should be higher than that used for classical textiles (10–30 µm) to enable adhesion and the efficient spreading of the osteoblast cell (>30 µm diameter). We report, for the first time, the fabrication of biodegradable nanostructured cylindrical PLLA (poly-L-Lactic acid) microfilaments of diameters 100 µm and 230 µm, using a single step melt-spinning process for straightforward integration of nano-scale ridge-like structures oriented in the fiber length direction. Appropriate drawing speed and temperature used during the filament spinning allowed for the creation of instabilities giving rise to nanofibrillar ridges, as observed by AFM (Atomic Force Microscopy). These micro-filaments were hydrophobic, and had reduced crystallinity and mechanical strength, but could still be processed into 2D/3D textile scaffolds of various shapes. Biological tests carried out on the woven scaffolds made from these nano-structured micro filaments showed excellent human bone cell MG 63 adhesion and proliferation, better than on smooth 30 µm- diameter fibers. Elongated filopodia of the osteoblast, intimately anchored to the nano-structured filaments, was observed. The filaments also induced in vitro osteogenic expression, as shown by the expression of osteocalcin and bone sialoprotein after 21 days of culture. This work deals with the fabrication of a new generation of nano-structured micro-filament for use as scaffolds of different shapes suited for bone cell engineering.

Influence of the Available Surface Area and Cell Elasticity on Bacterial Adhesion Forces on Highly Ordered Silicon Nanopillars.

Initial bacterial adhesion to solid surfaces is influenced by a multitude of different factors, e.g., roughness and stiffness, topography on the micro- and nanolevel, as well as chemical composition and wettability. Understanding the specific influences and possible interactive effects of all of these factors individually could lead to guidance on bacterial adhesion and prevention of unfavorable consequences like medically relevant biofilm formation. On this way, the aim of the present study was to identify the specific influence of the available surface area on the adhesion of clinically relevant bacterial strains with different membrane properties: Gram-positive Staphylococcus aureus and Gram-negative Aggregatibacter actinomycetemcomitans. As model surfaces, silicon nanopillar specimens with different spacings were fabricated using electron beam lithography and cryo-based reactive ion etching techniques. Characterization by scanning electron microscopy and contact angle measurement revealed almost defect-free highly ordered nanotopographies only varying in the available surface area. Bacterial adhesion forces to these specimens were quantified by means of single-cell force spectroscopy exploiting an atomic force microscope connected to a microfluidic setup (FluidFM). The nanotopographical features reduced bacterial adhesion strength by reducing the available surface area. In addition, the strain-specific interaction in detail depended on the bacterial cell’s elasticity and deformability as well. Analyzed by confocal laser scanning microscopy, the obtained results on bacterial adhesion forces could be linked to the subsequent biofilm formation on the different topographies. By combining two cutting-edge technologies, it could be demonstrated that the overall bacterial adhesion strength is influenced by both the simple physical interaction with the underlying nanotopography and its available surface area as well as the deformability of the cell.

Construction of a BMP-2 gene delivery system for polyetheretherketone bone implant material and its effect on bone formation in vitro.

Polyetheretherketone (PEEK) has been widely investigated for improving its biological inert to enable it to achieve stronger osteogenic capability and to be a promising material in implant fields. The most important mechanism that makes a successful implantation is osteointegration. Surface modification is an appropriate method to maintain the excellent mechanical properties of PEEK and simultaneously endow PEEK certain biological characters. In this work, we attempted to shape the nano-topography of PEEK surface by nitrogen low-temperature plasma and polydopamine coating on the surface as a secondary reaction platform to bond the aminated poly (lactic-co-glycolic acid) (PLGA) microspheres encapsulating the BMP-2 gene for enhancing the biological activity. Scanning electron microscope, atomic force microscopy, X-ray photoelectron spectroscopy and water contact angle (CA) measurements were applied to characterize the surface of modified or untreated PEEK. Surface characterization showed that the modification was successfully performed on PEEK including a rougher and more hydrophilic surface with nanotopographic features. The influence on cell adhesion, proliferation and differentiation was evaluated by culturing of rat bone marrow mesenchymal stem cells on different modified PEEK substrates in vitro. The biological results indicated that the low-temperature plasma treatment and PDA-coating on PEEK significantly promoted cell adhesion and proliferation. And the osteogenic differentiation was effectively improved by BMP-2 gene releasing from PLGA-NH2 microspheres. The results showed that this novel biological surface modification endowed PEEK with outstanding bioactivity and osteogenic ability, providing a theoretical basis for application in the field of implantation.

An update of nanotopographical surfaces in modulating stem cell fate: a narrative review.

ABSTRACTStem cells have been one of the ideal sources for tissue regeneration owing to their capability of self-renewal and differentiation. In vivo, the extracellular microenvironment plays a vital role in modulating stem cell fate. When developing biomaterials for regenerative medicine, incorporating biochemical and biophysical cues to mimic extracellular matrix can enhance stem cell lineage differentiation. More specifically, modulating the stem cell fate can be achieved by controlling the nanotopographic features on synthetic surfaces. Optimization of nanotopographical features leads to desirable stem cell functions, which can maximize the effectiveness of regenerative treatment. In this review, nanotopographical surfaces, including static patterned surface, dynamic patterned surface, and roughness are summarized, and their fabrication, as well as the impact on stem cell behaviour, are discussed. Later, the recent progress of applying nanotopographical featured biomaterials for altering different types of stem cells is presented, which directs the design and fabrication of functional biomaterial. Last, the perspective in fundamental research and for clinical application in this field is discussed.

Nanotopographic features induced by plasma flame modulate the osteoinductivity of titanium alloy

Nanotopographical surface modification of titanium alloy implants is effective in improving the osseointegration. Herein, a new nanotopography modulation approach was proposed based on the plasma flame (PF) generated by plasma spraying equipment. It was proved that the PF treatment can not only produce TiO2 on the Ti alloy surface, but also modulate its nanostructural characteristics. Besides a peak at 200–250 nm, the PF treatment induces extra light adsorption peaks located at 350–400 nm and 800–900 nm, depending on the nanostructure. In vitro cell culture experiments prove that the introduction of surface TiO2 nanotopography by the PF treatment modulates cell proliferation and osteogenic differentiation. This study highlights the potential of the PF treatment as a nanotopography modulator for enhancing the biological performance of the biomedical Ti alloy.

Cellular responses to nanoscale substrate topography of TiO2 nanotube arrays: cell morphology and adhesion.

Nanotopographical features can be beneficial in augmenting cell functions and increasing osteogenic potential. However, the relationships between surface topographies and biological responses are difficult to establish due to the difficulty in controlling the surface topographical features at a low-nanometre scale. Herein, we report the fabrication of well-defined controllable titanium dioxide (TiO2) nanotube arrays with a wide range of pore sizes, 30-175 nm in diameter, and use of the electrochemical anodization method to assess the effect of surface nanotopographies on cell morphology and adhesion. The results show that TiO2 nanotube arrays with pore sizes of 30 and 80 nm allowed for cell spreading of bone marrow-derived mesenchymal stem cells with increased cell area coverage. Additionally, cell adhesion was significantly enhanced by controlled nanotopographies of TiO2 nanotube arrays with 80 nm pore size. Our results demonstrate that surface modification at the nano-scale level with size tunability under controlled chemical/physical properties and culture conditions can greatly impact cell responses. These findings point to a new direction of material design for bone-tissue engineering in orthopaedic applications.

Hollow pollen grains as scaffolding building blocks in bone tissue engineering.

Introduction: The current study, for the first time, suggests nature-made pollen grains (PGs) of Pistacia vera L. as a potential candidate for using as scaffolding building blocks with encapsulation capability of bioactive compounds, such as bone morphogenetic protein 4 (BMP4). Methods: A modified method using KOH (5%, 25ºC) was developed to produce nonallergic hollow pollen grains (HPGs), confirmed by energy dispersive X-ray (EDX) analysis, field emission scanning electron microscopy (FESEM), and DNA and protein staining techniques. The in-vitro study was conducted on human adipose-derived mesenchymal stem cells (hAD-MSCs) to investigate the applicability of HPGs as bone scaffolding building blocks. Cytocompability was evaluated by FESEM, MTT assay, and gene expression analysis of apoptotic markers (BAX and BCL2). The osteoconductive potential of HPGs was assessed by alkaline phosphatase (ALP) activity measurement and gene expression analysis of osteogenic markers (RUNX2 and osteocalcin). Results: Findings demonstrated that HPGs can be considered as biocompatible compounds increasing the metabolic activities of the cells. Further, the bioactive nature of HPGs resulted in suitable cellular adhesion properties, required for a potent scaffold. The investigation of apoptotic gene expression indicated a reduced BAX/BCL2 ratio reflecting the protective effect of HPGs on hAD-MSCs. The increased ALP activity and expression of osteogenic genes displayed the osteoconductive property of HPGs. Moreover, the incorporation of BMP4 in HPGs initiated a synergistic effect on osteoblast maturation. Conclusion: Owing to the unique compositional and surface nanotopographical features of the Pistacia vera L. HPG, this microscale architecture provides a favorable microenvironment for the bottom-up remodeling of bone.

Heterogeneous Sorption of Radionuclides Predicted by Crystal Surface Nanoroughness.

Reactive transport modeling (RTM) is an essential tool for the prediction of contaminants' behavior in the bio- and geosphere. However, RTM of sorption reactions is constrained by the reactive surface site assessment. The reactive site density variability of the crystal surface nanotopography provides an "energetic landscape", responsible for heterogeneous sorption efficiency, not covered in current RTM approaches. Here, we study the spatially heterogeneous sorption behavior of Eu(III), as an analogue to trivalent actinides, on a polycrystalline nanotopographic calcite surface and quantify the sorption efficiency as a function of surface nanoroughness. Based on experimental data from micro-focus time-resolved laser-induced luminescence spectroscopy (μTRLFS), vertical scanning interferometry, and electron back-scattering diffraction (EBSD), we parameterize a surface complexation model (SCM) using surface nanotopography data. The validation of the quantitatively predicted spatial sorption heterogeneity suggests that retention reactions can be considerably influenced by nanotopographic surface features. Our study presents a way to implement heterogeneous surface reactivity into a SCM for enhanced prediction of radionuclide retention.

Thin films of functionalized carbon nanotubes support long-term maintenance and cardio-neuronal differentiation of canine induced pluripotent stem cells

Induced pluripotent stem cells (iPSCs) are a promising cell source for regenerative medicine. However, their feeder-free maintenance in undifferentiated states remains challenging. In recent past extensive studies have been directed using pristine or functionalized carbon nanotube in tissue engineering. Here we proposed thin films of functionalized carbon nanotubes (OH-single-walled CNTs [SWCNTs] and OH-multiwalled CNTs [MWCNTs]), as alternatives for the feeder-free in vitro culture of canine iPSCs (ciPSCs), considered as the cellular model. The ciPSC colonies could maintain their dome-shaped compactness and other characteristics when propagated on CNT films. Concomitantly, high cell viability and upregulation of pluripotency-associated genes and cell adhesion molecules were observed, further supported by molecular docking. Moreover, CNTs did not have profound toxic effects compared to feeder cultures as evident by cytocompatibility studies. Further, cardiac and neuronal differentiation of ciPSCs was induced on these films to determine their influence on the differentiation process. The cells retained differentiation potential and the nanotopographical features of the substrates provided positive cues to enhance differentiation to both lineages as evident by immunocytochemical staining and marker gene expression. Overall, OH-SWCNT provided better cues, maintained pluripotency, and induced the differentiation of ciPSCs. These results indicate that OH-functionalized CNT films could be used as alternatives for the feeder-free maintenance of ciPSCs towards prospective utilization in regenerative medicine.

Nanoconfinement of microvilli alters gene expression and boosts T cell activation.

T cells sense and respond to their local environment at the nanoscale by forming small actin-rich protrusions, called microvilli, which play critical roles in signaling and antigen recognition, particularly at the interface with the antigen presenting cells. However, the mechanism by which microvilli contribute to cell signaling and activation is largely unknown. Here, we present a tunable engineered system that promotes microvilli formation and T cell signaling via physical stimuli. We discovered that nanoporous surfaces favored microvilli formation and markedly altered gene expression in T cells and promoted their activation. Mechanistically, confinement of microvilli inside of nanopores leads to size-dependent sorting of membrane-anchored proteins, specifically segregating CD45 phosphatases and T cell receptors (TCR) from the tip of the protrusions when microvilli are confined in 200-nm pores but not in 400-nm pores. Consequently, formation of TCR nanoclustered hotspots within 200-nm pores allows sustained and augmented signaling that prompts T cell activation even in the absence of TCR agonists. The synergistic combination of mechanical and biochemical signals on porous surfaces presents a straightforward strategy to investigate the role of microvilli in T cell signaling as well as to boost T cell activation and expansion for application in the growing field of adoptive immunotherapy.

Hybrid interpenetrating hydrogel network favoring the bidirectional migration of tenocytes for rotator cuff tendon regeneration.

Replenishment of tenocytes to the injury site is an ideal strategy to improve healing response and accelerate the tendon ECM regeneration. The present study focused on the synthesis and characterization of a hybrid hydrogel scaffold system poly(propylene-fumarate)-alginate-polyvinyl alcohol-acrylic acid (PAPA) using poly(propylene-fumarate) (PPF), alginate, polyvinyl alcohol (PVA) and acrylic acid and the in vitro investigation of bidirectional mobility of swine shoulder tenocytes (SST) for its potential application in rotator-cuff tendon regeneration. IR analysis revealed the presence of alginate, PPF and PVA segments on the surface, SEM and AFM analyses revealed the porous and nano-topographical features of PAPA, respectively, swelling was 712.6 ± 84.21% with the EWC (%) of 87.59 ± 1.26 having the diffusional exponent and swelling constant 0.551 and 1.8, respectively. PAPA was biodegradable, cytocompatible and supported long-term survival of SSTs. SEM imaging revealed the adhesion, colonization, and sheet formation of SSTs within the PAPA hydrogel network. The SSTs seeded on the PAPA scaffolds were peculiar for their bidirectional migration as the anterograde movement was completed in 9 days whereas the retrograde infiltration occurred up to the depth of 198 μm. These findings suggest the promising translational potential of PAPA scaffold system in the management of rotator cuff tendon injury.