Search Results (17)

Anterior cruciate ligament (ACL) injuries pose significant challenges, driving the need for innovative repair strategies. Tissue engineering (TE) has emerged as a promising field for ACL injuries. Wet spinning is a filament production technique that enables precise control over filament alignment, diameter, and porosity, making it suitable for developing new scaffolds for ACL injuries. This study develops fibrous scaffolds using wet spinning of polycaprolactone (PCL) reinforced with cellulose nanocrystals (CNC) to enhance mechanical properties. Wet spinning was employed to fabricate fibrous scaffolds, utilizing PCL as the primary polymer due to its favorable biocompatibility and degradability. An automated collector was developed and optimized, which allowed the stretching of the PCL filaments to diameters as low as 30 µm. Several filaments were explored and characterized using SEM, TGA, and mechanical tests. The optimized PCL/CNC filaments were used to develop 3D braided structures that mimic ACL structure. The combination of wet-spun PCL/CNC filaments (with an optimized stirring method) and the braiding procedure allowed for fully biocompatible scaffolds that mimic both the structure and the mechanical properties of native ACL. Cytotoxicity and biocompatibility tests showed cell viability and proliferation values above 99% and 81%, respectively. These findings underscore the potential of CNC-reinforced PCL scaffolds as promising candidates for ACL repair, laying the groundwork for future biomedical applications. Full article

Using scaffolds is a promising alternative to current methods of treatment for ruptures of tendons and ligaments. However, scaffolds are subject to a wide range of challenges, including mechanical, degradation, process-related and biological requirements. Poly-ε-caprolactone (PCL) fibers have already shown potential for tendon tissue engineering (TTE) because of their degradation kinetics and excellent mechanical properties. The objective of this study was to enhance the PCL scaffold for TTE, specifically in regard to the filament morphology and collagen coating. PCL fibers were melt-spun as monofilaments with circular and snowflake-shaped cross-sections. Different scaffold densities were achieved by applying three different braiding angles in the braiding process. Morphological characterization was conducted including porosity and pore size distribution using µ-CT. The scaffolds were collagenized and cellularized with primary tenocytes for 7 days. Immunofluorescence staining showed a certain alignment of cell growing direction with fiber direction. In cell viability and cell proliferation assays, significant improvements in cell response were observed for the snowflake fiber and collagen coating groups, especially when combined. The data suggest that the utilization of non-circular fibers may facilitate enhanced cell guidance and surface area, while the application of a collagen coating could optimize the cellular environment for adhesion and proliferation. Full article

Injuries to tendons and ligaments are highly prevalent in the musculoskeletal system. Current treatments involve autologous transplants with limited availability and donor site morbidity. Tissue engineering offers a new approach through temporary load-bearing scaffolds. These scaffolds have to fulfill numerous requirements, the majority of which can be met using braiding combined with high-strength polycaprolactone (PCL) fibers. Considering regulatory requirements, several medical-grade PCL materials were assessed regarding their mechanical, degradational and cell biological properties. In the course of the investigation, an excellent fiber tensile strength of up to 850 MPa was achieved. The fibers were braided into multilayer scaffolds and scaled to match the human ACL. These were characterized regarding their morphology and their mechanical and degradational properties. Two strategies were followed to provide biological cues: (a) applying a chitosan-graft-PCL surface modification and (b) using non-circular fiber morphologies as topographical stimuli. Cell vitality assays showed generally positive cytocompatibility and no impairments due to the surface modification or material grade. The best cell vitality was achieved with a scaffold consisting of snowflake-shaped monofilaments combined with a 25° braiding angle. The surface modification equips the scaffold with a release platform for function molecules (as recently demonstrated) so that a holistic approach to addressing the numerous requirements is provided. Full article

The use of tissue engineering to address the shortcomings of current procedures for tendons and ligaments is promising, but it requires a suitable scaffold that meets various mechanical, degradation-related, scalability-related, and biological requirements. Macroporous textile scaffolds made from appropriate fiber material have the potential to fulfill the first three requirements. This study aimed to investigate the biocompatibility, sterilizability, and functionalizability of a multilayer braided scaffold. These macroporous scaffolds with dimensions similar to those of the human anterior cruciate ligament consist of fibers with appropriate tensile strength and degradation behavior melt-spun from Polycaprolactone (PCL). Two different cross-sectional geometries resulting in significantly different specific surface areas and morphologies were used at the fiber level, and a Chitosan-graft-PCL (CS-g-PCL) surface modification was applied to the melt-spun substrates for the first time. All scaffolds elicited a positive cell response, and the CS-g-PCL modification provided a platform for incorporating functionalization agents such as drug delivery systems for growth factors, which were successfully released in therapeutically effective quantities. The fiber geometry was found to be a variable that could be manipulated to control the amount released. Therefore, scaled, surface-modified textile scaffolds are a versatile technology that can successfully address the complex requirements of tissue engineering for ligaments and tendons, as well as other structures. Full article

The firm integration of anterior cruciate ligament (ACL) grafts into bones remains the most demanding challenge in ACL reconstruction, since graft loosening means graft failure. For a functional-tissue-engineered ACL substitute to be realized in future, robust bone attachment sites (entheses) have to be re-established. The latter comprise four tissue compartments (ligament, non-calcified and calcified fibrocartilage, separated by the tidemark, bone) forming a histological and biomechanical gradient at the attachment interface between the ACL and bone. The ACL enthesis is surrounded by the synovium and exposed to the intra-articular micromilieu. This review will picture and explain the peculiarities of these synovioentheseal complexes at the femoral and tibial attachment sites based on published data. Using this, emerging tissue engineering (TE) strategies addressing them will be discussed. Several material composites (e.g., polycaprolactone and silk fibroin) and manufacturing techniques (e.g., three-dimensional-/bio-printing, electrospinning, braiding and embroidering) have been applied to create zonal cell carriers (bi- or triphasic scaffolds) mimicking the ACL enthesis tissue gradients with appropriate topological parameters for zones. Functionalized or bioactive materials (e.g., collagen, tricalcium phosphate, hydroxyapatite and bioactive glass (BG)) or growth factors (e.g., bone morphogenetic proteins [BMP]-2) have been integrated to achieve the zone-dependent differentiation of precursor cells. However, the ACL entheses comprise individual (loading history) asymmetric and polar histoarchitectures. They result from the unique biomechanical microenvironment of overlapping tensile, compressive and shear forces involved in enthesis formation, maturation and maintenance. This review should provide a road map of key parameters to be considered in future in ACL interface TE approaches. Full article

The development of scaffolds for tissue-engineered growth of the anterior cruciate ligament (ACL) is a promising approach to overcome the limitations of current solutions. This work proposes novel biodegradable and biocompatible scaffolds matching the mechanical characteristics of the native human ligament. Poly(L-lactic acid) (PLA) scaffolds reinforced with graphite nano-platelets (PLA+EG) as received, chemically functionalized (PLA+f-EG), or functionalized and decorated with silver nanoparticles [PLA+((f-EG)+Ag)], were fabricated by conventional braiding and using 3D-printing technology. The dimensions of both braided and 3D-printed scaffolds were finely controlled. The results showed that the scaffolds exhibited high porosity (>60%), pore interconnectivity, and pore size suitable for ligament tissue ingrowth, with no relevant differences between PLA and composite scaffolds. The wet state dynamic mechanical analysis at 37 °C revealed an increase in the storage modulus of the composite constructs, compared to neat PLA scaffolds. Either braided or 3D-printed scaffolds presented storage modulus values similar to those found in soft tissues. The tailorable design of the braided structures, as well as the reproducibility, the high speed, and the simplicity of 3D-printing allowed to obtain two different scaffolds suitable for ligament tissue engineering. Full article

The anterior cruciate ligament (ACL) is the most commonly injured intra-articular ligament of the knee. Due to its limited intrinsical healing potential and vascularization, injuries of the ACL do not heal satisfactorily, and surgical intervention is usually required. The limitations of existing reconstructive grafts and autologous transplants have prompted interest in tissue-engineered solutions. A tissue engineering scaffold for ACL reconstruction must be able to mimic the mechanical properties of the native ligament, provide sufficient porosity to promote cell growth of the neoligament tissue, and be biodegradable. This study investigates long-term biodegradable poly-ε-caprolactone (PCL)-based scaffolds for ACL replacement using the 3D hexagonal braiding technique. The scaffolds were characterized mechanically as well as morphologically. All scaffolds, regardless of their braid geometry, achieved the maximum tensile load of the native ACL. The diameter of all scaffolds was lower than that of the native ligament, making the scaffolds implantable with established surgical methods. The 3D hexagonal braiding technique offers a high degree of geometrical freedom and, thus, the possibility to develop novel scaffold architectures. Based on the findings of this study, the 3D-braided PCL-based scaffolds studied were found to be a promising construct for tissue engineering of the anterior cruciate ligament. Full article

Anterior Cruciate Ligament (ACL) injuries are becoming more prevalent in athletes. Anterior Cruciatum Ligament Reconstruction (ACLR) surgery was used to treat ACL injuries and resulted in a recurrence rate of 94% due to the biomechanically repaired tissue being weaker than the original tissue. As a result, biodegradable artificial ligaments must be developed that can withstand mechanical stress during neoligament formation and stabilize the ACL. The purpose of this study is to determine the effect of composition variations in polylactic acid (PLA) and polycaprolactone (PCL) used as ACL nanofiber scaffolds on ultimate tensile strength (UTS) and modulus of elasticity, fiber diameter, cytotoxicity level, and degradation level, as well as the PLA-PCL concentration that provides the best value as an ACL scaffold. Electrospinning was used to fabricate the nanofiber scaffold with the following PLA-PCL compositions: A (100:0), B (85:15), C (80:20), D (70:30), and E (0:100) (wt%). The functional group test revealed no new peaks in any of the samples, and the ester group could be identified in the C-O bond at wave numbers 1300–1100 cm⁻¹ and in the C=O bond at wave numbers 1750–1730 cm⁻¹. The average fiber diameter, as determined by SEM morphology, is between 1000 and 2000 nm. The unbraided sample had a UTS range of 1.578–4.387 MPa and an elastic modulus range of 8.351–141.901 MPa, respectively, whereas the braided sample had a range of 0.879–1.863 MPa and 2.739–4.746 MPa. The higher the PCL composition, the lower the percentage of viable cells and the faster the sample degrades. All samples had a cell viability percentage greater than 60%, and samples C, D, and E had a complete degradation period greater than six months. The ideal scaffold, Sample C, was composed of PLA-PCL 80:20 (wt%), had an average fiber diameter of 827 ± 271 nm, a living cell percentage of 97.416 ± 5.079, and a degradation time of approximately 219 days. Full article

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. Full article

As the incidence of cardiovascular diseases has been growing in recent years, the need for small-diameter vascular grafts is increasing. Considering the limited success of synthetic grafts, vascular tissue engineering/repair/regeneration aim to find novel solutions. Silk fibroin (SF) has been widely investigated for the development of vascular grafts, due to its good biocompatibility, tailorable biodegradability, excellent mechanical properties, and minimal inflammatory reactions. In this study, a new generation of three-layered SF vascular scaffolds has been produced and optimized. Four designs of the SILKGraft vascular prosthesis have been developed with the aim of improving kink resistance and mechanical strength, without compromising the compliance with native vessels and the proven biocompatibility. A more compact arrangement of the textile layer allowed for the increase in the mechanical properties along the longitudinal and circumferential directions and the improvement of the compliance value, which approached that reported for the saphenous and umbilical veins. The higher braid density slightly affected the grafts’ morphology, increasing surface roughness, but the novel design mimicked the corrugation approach used for synthetic grafts, causing significant improvements in kink resistance. Full article

High-performance fibroin fibres are ideal candidates for the manufacture of scaffolds with applications in tissue engineering due to the excellent mechanical properties and optimal biocompatibility of this protein. In this work, the manufacture of high-strength fibres made from the silk glands of Samia cynthia ricini is explored. The glands were subjected to soaking in aqueous dissolutions of acetic acid and stretched to manufacture the fibres. The materials produced were widely characterized, in terms of morphology, mechanical properties, crystallinity and content of secondary structures, comparing them with those produced by the standard procedure published for Bombyx mori. In addition, mechanical properties and biocompatibility of a braided scaffold produced from these fibres was evaluated. The results obtained show that the fibres from B. mori present a higher degree of crystallinity than those from S. c. ricini, which is reflected in higher values of elastic modulus and lower values of strain at break. Moreover, a decrease in the elongation values of the fibres from S. c. ricini was observed as the concentration of acetic acid was increased during the manufacture. On the other hand, the study of the braided scaffolds showed higher values of tensile strength and strain at break in the case of S. c. ricini materials and similar values of elastic modulus, compared to those of B. mori, displaying both scaffolds optimal biocompatibility using a fibroblast cell line. Full article

Tissue Engineering is considered a promising route to address existing deficits of autografts and permanent synthetic prostheses for tendons and ligaments. However, the requirements placed on the scaffold material are manifold and include mechanical, biological and degradation-related aspects. In addition, scalable processes and FDA-approved materials should be applied to ensure the transfer into clinical practice. To accommodate these aspects, this work focuses on the high-scale fabrication of high-strength and highly oriented polycaprolactone (PCL) fibers with adjustable cross-sectional geometry and degradation kinetics applying melt spinning technology. Four different fiber cross-sections were investigated to account for potential functionalization and cell growth guidance. Mechanical properties and crystallinity were studied for a 24-week exposure to phosphate-buffered saline (PBS) at 37 °C. PCL fibers were further processed into scaffolds using multistage circular braiding with three different hierarchical structures. One structure was selected based on its morphology and scaled up in thickness to match the requirements for a human anterior cruciate ligament (ACL) replacement. Applying a broad range of draw ratios (up to DR9.25), high-strength PCL fibers with excellent tensile strength (up to 69 cN/tex) could be readily fabricated. The strength retention after 24 weeks in PBS at 37 °C was 83–93%. The following braiding procedure did not affect the scaffolds’ mechanical properties as long as the number of filaments and the braiding angle remained constant. Up-scaled PCL scaffolds resisted loads of up to 4353.88 ± 37.30 N, whilst matching the stiffness of the human ACL (111–396 N/mm). In conclusion, this work demonstrates the fabrication of highly oriented PCL fibers with excellent mechanical properties. The created fibers represent a promising building block that can be further processed into versatile textile implants for tissue engineering and regenerative medicine. Full article

(1) Background: A suitable scaffold with adapted mechanical and biological properties for ligament tissue engineering is still missing. (2) Methods: Different scaffold configurations were characterized in terms of morphology and a mechanical response, and their interactions with two types of stem cells (Wharton’s jelly mesenchymal stromal cells (WJ-MSCs) and bone marrow mesenchymal stromal cells (BM-MSCs)) were assessed. The scaffold configurations consisted of multilayer braids with various number of silk layers (n = 1, 2, 3), and a novel composite scaffold made of a layer of copoly(lactic acid-co-(e-caprolactone)) (PLCL) embedded between two layers of silk. (3) Results: The insertion of a PLCL layer resulted in a higher porosity and better mechanical behavior compared with pure silk scaffold. The metabolic activities of both WJ-MSCs and BM-MSCs increased from day 1 to day 7 except for the three-layer silk scaffold (S3), probably due to its lower porosity. Collagen I (Col I), collagen III (Col III) and tenascin-c (TNC) were expressed by both MSCs on all scaffolds, and expression of Col I was higher than Col III and TNC. (4) Conclusions: the silk/PLCL composite scaffolds constituted the most suitable tested configuration to support MSCs migration, proliferation and tissue synthesis towards ligament tissue engineering. Full article

We developed a (three-dimensional) 3D scaffold, we named HY-FIB, incorporating a force-transmission band of braided hyaluronate embedded in a cell localizing fibrin hydrogel and poly-lactic-co-glycolic acid (PLGA) nanocarriers as transient components for growth factor controlled delivery. The tenogenic supporting capacity of HY-FIB on human-Bone Marrow Mesenchymal Stem Cells (hBM-MSCs) was explored under static conditions and under bioreactor-induced cyclic strain conditions. HY-FIB elasticity enabled to deliver a mean shear stress of 0.09 Pa for 4 h/day. Tendon and cytokine marker expression by hBM-MSCs were studied. Results: hBM-MSCs embedded in HY-FIB and subjected to mechanical stimulation, resulted in a typical tenogenic phenotype, as indicated by type 1 Collagen fiber immunofluorescence. RT-qPCR showed an increase of type 1 Collagen, scleraxis, and decorin gene expression (3-fold, 1600-fold, and 3-fold, respectively, at day 11) in dynamic conditions. Cells also showed pro-inflammatory (IL-6, TNF, IL-12A, IL-1β) and anti-inflammatory (IL-10, TGF-β1) cytokine gene expressions, with a significant increase of anti-inflammatory cytokines in dynamic conditions (IL-10 and TGF-β1 300-fold and 4-fold, respectively, at day 11). Mechanical signaling, conveyed by HY-FIB to hBM-MSCs, promoted tenogenic gene markers expression and a pro-repair cytokine balance. The results provide strong evidence in support of the HY-FIB system and its interaction with cells and its potential for use as a predictive in vitro model. Full article

Hydroxyapatite (HA) coating is successfully prepared by electrodeposition on the surface of polyvinyl alcohol (PVA)/polylactic acid (PLA) braid which serves as a potential biodegradable bone scaffold. The surface morphology, element composition, crystallinity and chemical bonds of HA coatings at various deposition times (60, 75, 90, 105 and 120 min) are characterized by scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR), respectively. Average Surface roughness (Ra) of HA coating is observed by confocal microscopy. The results reveal that the typical characteristic peaks of the FTIR spectrum confirm that HA coating is successfully prepared on the rugged surface of the PVA/PLA braid. The XRD results indicate that the crystallinity of HA can be improved by increasing deposition time. In the 90 min-deposition, hydroxyapatite has a dense and uniform coating morphology, Ca/P ratio of 1.7, roughness of 0.725 μm, which shows the best electrodeposition performance. The formation mechanism of granular and plate-like hydroxyapatite crystals is explained by the structural characteristics of a hydroxyapatite unit cell. This study provides a foundation for a bone scaffold braided by biodegradable fibers. Full article