Porous Polycaprolactone Scaffolds Research Articles

Growth and endothelial differentiation of adipose stem cells on polycaprolactone

Adipose tissue is a readily available source of multipotent adult stem cells for use in tissue engineering/regenerative medicine. Various growth factors have been used to stimulate acquisition of endothelial characteristics by adipose-derived stem cells (ADSC). Herein, we study the growth and endothelial differentiation potential of ADSC seeded onto a porous polycaprolactone (PCL) scaffold. The objective of this study is to demonstrate that PCL is a good material to be used as a scaffold to support reconstruction of new endothelial tissue using adipose stem cells. We found that undifferentiated ADSC adhere and grow on PCL. We show that, after culture in endothelial differentiation medium, ADSC were positive to LDL uptake and expressed molecular markers characteristic of endothelial cells (CD31; eNOS and vWF). In addition, our study defines the time required for the differentiation of ADSC directly onto PCL. This study suggests that PCL can be used as a scaffold to generate endothelial tissue in vitro. PLC has excellent mechanical properties and a slow degradation rate. Moreover, based on our results, we propose that PCL could be used to graft scaffolds coated with endothelial cells derived from ADSC stem cells. Endothelial cells-coated PCL could find several applications to replace damaged area of the body; for example, a possible use could be the generation of vascular grafts.

Nanoshell assisted laser soldering of vascular tissue

Laser tissue soldering (LTS) is a promising technique for tissue fusion but is limited by the lack of reproducibility particularly when the amount of indocyanine green (ICG) applied as energy absorber cannot be controlled during the soldering procedure. Nanotechnology enables the control over the quantitative binding of the ICG. The aim of this study was to establish a highly reproducible and strong tissue fusion using ICG packed nanoshells. By including the chromophore in the soldering scaffold, dilution of the energy absorber during the soldering procedure is prevented. The feasibility of this novel nanoshell soldering technique was studied by assessing the local heating of the area and tensile strength of the resulting fused tissue. Nanoshells with a diameter of 250-270 nm were loaded with ICG and included in a porous polycaprolactone (PCL) scaffold doped with albumin solder. The nanoshell scaffold was used in a flexible, semi-dry formulation suitable for surgical use. Heat development, tensile strength as well as tissue damage were assessed. Rabbit aortic arteries were successfully soldered using an ICG packed nanoshell scaffold. Tensile strengths of these nanoshell soldered anastomoses were found to be 734 ± 327 mN (median = 640 mN). Thermal damage was restricted to the adventitia at the irradiated area. In addition, absorber dilution was prevented during the soldering procedure resulting in significantly lower variance in maximum temperature (P = 0.03) compared to the classical liquid ICG soldering technique. Using nanoshells, controlled amounts of chromophore could successfully be bound into the polymer scaffold. Diode laser soldering of vascular tissue using ICG-nanoshell scaffolds leads to strong and reproducible tissue fusion. With optimally chosen settings of irradiation time, nanoshells coating and scaffold properties, our improved LTS procedure demonstrates the potential for a clinically applicable anastomosis technique.

Visualization of the Penetration Depth of Plasma in Three-Dimensional Porous PCL Scaffolds

A dielectric barrier discharge (DBD) discharge is used to modify the surface properties of 3-D porous polycaprolactone (PCL) scaffolds. After plasma treatment, the penetration of blue ink into the samples was used to determine the effectiveness of the plasma treatment inside the structures. It was found that the ink could penetrate deeper into the scaffolds after plasma treatment.

A novel nano-structured porous polycaprolactone scaffold improves hyaline cartilage repair in a rabbit model compared to a collagen type I/III scaffold: in vitro and in vivo studies

To develop a nano-structured porous polycaprolactone (NSP-PCL) scaffold and compare the articular cartilage repair potential with that of a commercially available collagen type I/III (Chondro-Gide) scaffold. By combining rapid prototyping and thermally induced phase separation, the NSP-PCL scaffold was produced for matrix-assisted autologous chondrocyte implantation. Lyophilizing a water-dioxane-PCL solution created micro and nano-pores. In vitro: The scaffolds were seeded with rabbit chondrocytes and cultured in hypoxia for 6 days. qRT-PCR was performed using primers for sox9, aggrecan, collagen type 1 and 2. In vivo: 15 New Zealand White Rabbits received bilateral osteochondral defects in the femoral intercondylar grooves. Autologous chondrocytes were harvested 4 weeks prior to surgery. There were 3 treatment groups: (1) NSP-PCL scaffold without cells. (2) The Chondro-Gide scaffold with autologous chondrocytes and (3) NSP-PCL scaffold with autologous chondrocytes. Observation period was 13 weeks. Histological evaluation was made using the O'Driscoll score. In vitro: The expressions of sox9 and aggrecan were higher in the NSP-PCL scaffold, while expression of collagen 1 was lower compared to the Chondro-Gide scaffold. In vivo: Both NSP-PCL scaffolds with and without cells scored significantly higher than the Chondro-Gide scaffold when looking at the structural integrity and the surface regularity of the repair tissue. No differences were found between the NSP-PCL scaffold with and without cells. The NSP-PCL scaffold demonstrated higher in vitro expression of chondrogenic markers and had higher in vivo histological scores compared to the Chondro-Gide scaffold. The improved chondrocytic differentiation can potentially produce more hyaline cartilage during clinical cartilage repair. It appears to be a suitable cell-free implant for hyaline cartilage repair and could provide a less costly and more effective treatment option than the Chondro-Gide scaffold with cells.

Accelerating Vascularization in Polycaprolactone Scaffolds by Endothelial Progenitor Cells

Vascularization is a major challenge in tissue engineering. The purpose of this study is to expedite the formation of blood vessels in porous polycaprolactone (PCL) scaffolds by the delivery of endothelial progenitor cells (EPCs). To establish a pro-angiogenic and pro-vasculogenic microenvironment, we employed EPCs seeded in PCL scaffold with surface-immobilized heparin and vascular endothelial growth factor (VEGF). EPCs seeded on scaffolds with VEGF exhibited phosphorylation of the receptor. After 7 days of subcutaneous implantation in immunodeficient mice, heparin-immobilized PCL scaffolds with VEGF induced significantly high density of blood vessel formation. The anastomosis of EPC-derived vessels with the host circulatory system was evident by the presence of murine erythrocytes in the lumen of human-CD31 positive vessels. A more uniform distribution of blood vessels was achieved within 2-mm thick scaffolds by seeding an optimal density of EPCs. The seeding of a higher density of EPC resulted in an increase in apoptosis and a concomitant decline in blood vessel formation at the scaffold's inner core. When co-seeded with other cells, the EPCs maintained the ability to accelerate vessel formation. The excessive expansion of EPCs in vitro was associated with a decline in their in vivo vasculogenic potential. EPCs accelerated the vascularization of heparin-immobilized PCL scaffolds in the presence of VEGF.

Fabrication of tissue engineered PCL scaffold by selective laser-sintered machine for osteogeneisis of adipose-derived stem cells

Polycaprolactone (PCL) is a bioresorbable polymer with potential applications for bone and cartilage repair. In this work, three-dimensional (3D) and porous PCL scaffolds were designed and fabricated via selective laser sintering (SLS). The aim of this study was to evaluate the osteogenic potential of porcine adipose-derived stem cells (pASCs) in a laser-sintered PCL (lsPCL) scaffold. The character of the lsPCL scaffold was evaluated. The pore size and the microstructure were observed by SEM. The pASCs were harvested and isolated from pig inguinal area. Then, the lsPCL scaffold was seeded with ASCs and cultured in osteogenic medium for 0 and 14 days. Cell proliferation was measured by MTS. Alkaline phosphatase activity (ALP) was detected using biochemical methods. SEM was used to observe the interaction between scaffold and cell. An energy dispersive spectrum (EDS) was used to analyze the mineralization in each group. Porosity was around 83%; pore size was around 300–400 µm. Both MTS and ALP showed significant increase after subcultivation in osteogenic medium for 14 days. SEM detailed that the pASCs cell can attach well to the lsPCL scaffold. The energy dispersive spectrum (EDS) also demonstrated calcium deposits around pASCs after osteo-induction for 14 days. In contrast, no mineralization was found around ASCs after osteo-induction of 0 days. In conclusion, the laser-sintered PCL is a suitable scaffold for the proliferation of ASCs. The ASCs were also well differentiated into osteoblasts in the 3D, porous, laser-sintered PCL scaffold.

The enhancement of VEGF-mediated angiogenesis by polycaprolactone scaffolds with surface cross-linked heparin

This study investigates the effect of surface cross-linked heparin on vascular endothelial growth factor (VEGF)-mediated angiogenesis in porous polycaprolactone (PCL) scaffolds in vivo. We tested the hypothesis that VEGF delivered by scaffolds coated with a sub-micron thick layer of immobilized heparin would accelerate angiogenesis. The bioactivity of retained VEGF was confirmed by its phosphorylation of VEGF receptor-2. After 7 and 14 days of subcutaneous implantation in mice, the heparin-PCL scaffolds loaded with VEGF displayed significantly higher infiltration of blood vessels which traversed the entire scaffold thickness (2 mm). The stability and function of the newly formed vessels were confirmed by smooth muscle cell coverage and vessel perfusability, respectively. The contribution of individual components was assessed by varying the VEGF dose and heparin thickness. Prolonging the cross-linking reaction on PCL scaffolds resulted in higher heparin content, thicker heparin layer, and higher VEGF retention. While a dose dependent angiogenic response was observed with VEGF, higher amount of cross-linked heparin did not translate into additional improvement in angiogenesis for a given dose of VEGF. The synergism of immobilized heparin and VEGF in stimulating angiogenesis was observed in vivo.

Mechanical and microstructural properties of polycaprolactone scaffolds with one-dimensional, two-dimensional, and three-dimensional orthogonally oriented porous architectures produced by selective laser sintering

This article reports on the experimental determination and finite element modeling of tensile and compressive mechanical properties of solid polycaprolactone (PCL) and of porous PCL scaffolds with one-dimensional, two-dimensional and three-dimensional orthogonal, periodic porous architectures produced by selective laser sintering (SLS). PCL scaffolds were built using optimum processing parameters, ensuring scaffolds with nearly full density (>95%) in the designed solid regions and with excellent geometric and dimensional control (within 3–8% of design). The tensile strength of bulk PCL ranged from 10.5 to 16.1 MPa, its modulus ranged from 343.9 to 364.3 MPa, and the tensile yield strength ranged from 8.2 to 10.1 MPa. These values are consistent with reported literature values for PCL processed through various manufacturing methods. Across porosity ranged from 56.87% to 83.3%, the tensile strength ranged from 4.5 to 1.1 MPa, the tensile modulus ranged from 140.5 to 35.5 MPa, and the yield strength ranged from 3.2 to 0.76 MPa. The compressive strength of bulk PCL was 38.7 MPa, the compressive modulus ranged from 297.8 to 317.1 MPa, and the compressive yield strength ranged from 10.3 to 12.5 MPa. Across porosity ranged from 51.1% to 80.9%, the compressive strength ranged from 10.0 to 0.6 MPa, the compressive modulus ranged from 14.9 to 12.1 MPa, and the compressive yield strength ranged from 4.25 to 0.42 MPa. These values, while being in the lower range of reported values for trabecular bone, are the highest reported for PCL scaffolds produced by SLS and are among the highest reported for similar PCL scaffolds produced through other layered manufacturing techniques. Finite element analysis showed good agreement between experimental and computed effective tensile and compressive moduli. Thus, the construction of bone tissue engineering scaffolds endowed with oriented porous architectures and with predictable mechanical properties through SLS is demonstrated.

Porous polycaprolactone scaffold for cardiac tissue engineering fabricated by selective laser sintering

An advanced manufacturing technique, selective laser sintering (SLS), was utilized to fabricate a porous polycaprolactone (PCL) scaffold designed with an automated algorithm in a parametric library system named the “computer-aided system for tissue scaffolds” (CASTS). Tensile stiffness of the sintered PCL strut was in the range of 0.43 ± 0.15 MPa when a laser power of 3 W and scanning speed of 150 in s −1 was used. A series of compressive mechanical characterizations was performed on the parametric scaffold design and an empirical formula was presented to predict the compressive stiffness of the scaffold as a function of total porosity. In this work, the porosity of the scaffold was selected to be 85%, with micropores (40–100 μm) throughout the scaffold. The compressive stiffness of the scaffold was 345 kPa. The feasibility of using the scaffold for cardiac tissue engineering was investigated by culturing C2C12 myoblast cells in vitro for 21 days. Fluorescence images showed cells were located throughout the scaffold. High density of cells at 1.2 × 10 6 cells ml −1 was recorded after 4 days of culture. Fusion and differentiation of C2C12 were observed as early as 6 days in vitro and was confirmed with myosin heavy chain immunostaining after 11 days of cell culture. A steady population of cells was then maintained throughout 21 days of culturing. This work demonstrated the feasibility of tailoring the mechanical property of the scaffold for soft tissue engineering using CASTS and SLS. The macroarchitecture of the scaffold can be modified efficiently to fabricate scaffolds with different macropore sizes or changing the elemental cell design in CASTS. Further process and design optimization could be carried out in the future to fabricate scaffolds that match the tensile strength of native myocardium, which is of the order of tens of kPa.

The interaction between bone marrow stromal cells and RGD-modified three-dimensional porous polycaprolactone scaffolds

We previously established a simple method to immobilize the Arg-Gly-Asp (RGD) peptide on polycaprolactone (PCL) two-dimensional film surfaces that significantly improved bone marrow stromal cell (BMSC) adhesion to these films. The current work extends this modification strategy to three-dimensional (3D) PCL scaffolds to investigate BMSC attachment, cellular distribution and cellularity, signal transduction and survival on the modified PCL scaffold compared to those on the untreated ones. The results demonstrated that treatment of 3D PCL scaffold surfaces with 1,6-hexanediamine introduced the amino functional groups onto the porous PCL scaffold homogenously as detected by a ninhydrin staining method. Followed by the cross-linking reaction, RGDC peptide was successfully immobilized on the surface of PCL scaffold. Although the static seeding method used in this study caused heterogeneous cell distribution, the RGD-modified PCL scaffold still demonstrated the improved BMSC attachment and cellular distribution in the scaffold. More importantly, the integrin-mediated signal transduction FAK–PI3K–Akt pathway was significantly up-regulated by RGD modification and a subsequent increase in cell survival and growth was found in the modified scaffold. The present study introduces an easy method to immobilize RGD peptide on the 3D porous PCL scaffold and provides further evidence that modification of 3D PCL scaffolds with RGD peptides elicits specific cellular responses and improves the final cell–biomaterial interaction.

Local Thickness and Anisotropy Approaches to Characterize Pore Size Distribution of Three-Dimensional Porous Networks

Two image-analysis approaches for pore size distribution (PSD) of porous media are proposed. The methods are based on the skeleton representation of a porous object. One approach gives the local thickness of the pore object to represent the pore size corresponding to a lower limit of PSD. The other gives the pore size taking into account the anisotropy of pore object and corresponds to an upper limit of PSD. These two approaches can be incorporated into a computer program without computationally intensive and complex mathematical operations. In this study, these two approaches are applied to a two-dimensional (2D) synthetic image and 3D natural images of tissue scaffolds with various porosities and tortuosities. The scaffolds were prepared by removing the water-soluble poly(ethylene oxide) (PEO) component of the polycaprolactone (PCL)/PEO blend, leaving a porous PCL scaffold. Extracting quantitative PSD information for materials with an interconnected porous network rather than discrete voids (such as tissue scaffolds) is inevitably subjective without a universally accepted definition of "pore size." Therefore, the proposed lower and upper limits of PSD can come into play when considering mass transfer and scaffold surface area for cell-matrix interaction.

Solder doped polycaprolactone scaffold enables reproducible laser tissue soldering

In this in vitro feasibility study we analyzed tissue fusion using bovine serum albumin (BSA) and Indocyanine green (ICG) doped polycaprolactone (PCL) scaffolds in combination with a diode laser as energy source while focusing on the influence of irradiation power and albumin concentration on the resulting tensile strength and induced tissue damage. A porous PCL scaffold doped with either 25% or 40% (w/w) of BSA in combination with 0.1% (w/w) ICG was used to fuse rabbit aortas. Soldering energy was delivered through the vessel from the endoluminal side using a continuous wave diode laser at 808 nm via a 400 microm core fiber. Scaffold surface temperatures were analyzed with an infrared camera. Optimum parameters such as irradiation time, radiation power and temperature were determined in view of maximum tensile strength but simultaneously minimum thermally induced tissue damage. Differential scanning calorimetry (DSC) was performed to measure the influence of PCL on the denaturation temperature of BSA. Optimum parameter settings were found to be 60 seconds irradiation time and 1.5 W irradiation power resulting in tensile strengths of around 2,000 mN. Corresponding scaffold surface temperature was 117.4+/- 12 degrees C. Comparison of the two BSA concentration revealed that 40% BSA scaffold resulted in significant higher tensile strength compared to the 25%. At optimum parameter settings, thermal damage was restricted to the adventitia and its interface with the outermost layer of the tunica media. The DSC showed two endothermic peaks in BSA containing samples, both strongly depending on the water content and the presence of PCL and/or ICG. Diode laser soldering of vascular tissue using BSA-ICG-PCL-scaffolds leads to strong and reproducible tissue bonds, with vessel damage limited to the adventitia. Higher BSA content results in higher tensile strengths. The DSC-measurements showed that BSA denaturation temperature is lowered by addition of water and/or ICG-PCL.

In vitro mineralization of surface-modified porous polycaprolactone scaffolds in simulated body fluid

Porous polycaprolactone (PCL) scaffolds were fabricated by combination of porogen-leaching and freeze-drying processes. Ice particulates were used as porogen materials. The porous PCL scaffolds were modified by potassium hydroxide solution with concentration of 1 mol/L at room temperature for 8 h, subsequently biomineralized in simulated body fluid for 2 h and 8 h, respectively. The microstructure and characteristics of the PCL scaffolds were investigated by scanning electron microscope (SEM) and EDS. The results showed (1) PCL scaffolds had high degree of connectivity and different pore sizes. (2) Plate-like apatite was observed on the surface of the scaffolds after being immersed into SBF for 8 h.

Preparation of Porous Polycaprolactone Scaffolds by Using Freeze-Drying Combined Porogen-Leaching Methods

The microstructure of scaffold was one of key factors for tissue engineering. Porous polycaprolactone (PCL) scaffolds were fabricated by combination of porogen-leaching and freeze-drying process. Ice particulates were used as porogen material, and PCL solutions in chloroform were mixed with ice particulates for 5minuture at zero temperature. Then the mixture was freezed in liquid nitrogen, and porous scaffold was prepared by freeze - drying finally. The microstructure and properties of the scaffolds were investigated. Porous structure of the scaffolds showed that good 3D microstructure and no porogen remained in the scaffold; pore size and porosity were determined by the size and mass fraction of ice particulates. The results demonstrated that the Scaffolds possessed open and interconnected pores with sizes ranging from several μm to more than 300μm and porosities of 50~80%.

A porous PCL scaffold promotes the human chondrocytes redifferentiation and hyaline‐specific extracellular matrix protein synthesis

The redifferentiation, proliferation, and hyaline-specific extracellular matrix (ECM) protein synthesis of chondrocytes cultured in a polycaprolactone (PCL) scaffold were analyzed. Gene expression of the type II collagen and aggrecan was assessed by real-time PCR in cells from PCL scaffolds, monolayer, and pellet cultures. The proliferative activity was assessed using Ki-67 immunodetection, and the chondrocytic differentiation was evaluated using S-100 immunodetection. The synthesis and deposition into scaffold pores of type II collagen and glycosaminoglycan were analyzed by immunohistochemistry and Alcian blue staining, respectively. All parameters were assessed throughout 28 days of cultures maintained in either fetal bovine serum-containing medium (FCM) or Insulin-Transferrin-Selenium-containing medium (ICM). Expression of the type II collagen gene was lower in FCM cultures than in ICM cultures for all culture systems (p < 0.05). Moreover, PCL scaffolds cultured in ICM were able to induce collagen gene expression more efficiently than pellet and monolayer cultures. Aggrecan gene expression did not vary significantly between mediums and three-dimensional system cultures, but in ICM cultures, the monolayer cultures had significantly higher levels of aggrecan gene expression than did either the PCL or pellet cultures. Chondrocytes cultured in PCL scaffolds or pellets with FCM did not proliferate to a great extent but did maintain their differentiated phenotype for 28 days. Levels of cartilage ECM protein synthesis and deposition into the scaffold pores were similar among PCL and pellet cultures grown in FCM and in ICM. In conclusion, chondrocytes seeded into PCL scaffolds, cultured in ICM, efficiently maintained their differentiated phenotype and were able to synthesize cartilage-specific ECM proteins.

Porous Polymer Prosthesis for Meniscal Regeneration

Meniscus is the most commonly injured structure in the knee joint. Resection of the meniscus as well as the torn menisci is known to induce the degeneration of the articular cartilage. Replacement of the resected meniscus by allograft is limited by its availability and potential disease transmission. Artificial prostheses are being tried in an attempt to regenerate the meniscal tissue and we developed the biodegradable porous polycaprolactone(PCL) scaffold, which acts as a temporary scaffold to enable the regeneration of a new tissue in time. We report the results of rabbit implantation model. Biodegradable PCL scaffold coated with type I collagen with pores sized 100~150 +m and with compression modulus 400 kpa were fabricated by melt-molding particulate- leaching method. The molds were made using the native meniscus of the rabbit. Medial meniscus of right knee was partially removed through arthrotomy, leaving anterior 1/5 of the meniscus, after sectioning medial collateral ligament. The implant was attached to the peripheral capsule and remnant anterior meniscus with sutures. The medial meniscus of the left knee was removed and served as a control without replacement. The regenerated meniscus was harvested at 4 &amp; 12wks after implantation. In addition to the routine histology of the tissue regenerated and remnant scaffold, junction between the normal meniscus and the regenerated tissue, and cartilage surface degeneration was observed. After 4 and 12 weeks the scaffolds, although considerable amount of the materials remained, were largely filled and covered with fibrous tissue which was assumed to be derived from synovial tissue of peripheral capsule. The tissue grossly resembling the native meniscus was maintained and spindle shaped cells with extracellular matrices were observed histologically. Neither cells with chondrocytic phenotype nor distinct cartilage matrices were observed until 12 weeks. The bonding between the regenerated tissue and the peripheral synovial capsule was firm and solid in all cases. The tissue bridges between the native meniscus and the regenerated tissue were found in 9/10 operated knees. Articular surface degeneration was not different between experimental and control groups except one case. More or less, the extrusion of the meniscus was found in almost all knees. This study revealed that meniscal replacement with PCL polymer prosthesis was feasible and led to adequate tissue formation. Long term studies on adaptive remodeling will be required.

Biomineralization of calcium disilicide in porous polycaprolactone scaffolds

The incorporation of CaSi(2) grains within a polycaprolactone (PCL) framework results in bioactive and biodegradable scaffolds which may be used in bone tissue regeneration. Porous PCL scaffolds were prepared via a combination of salt-leaching and microemulsion methods. To provide markedly different structural environments for the inorganic phase, calcium disilicide powder was either added to a mixed-composition porogen during a given scaffold preparation, or alternatively added to pre-formed scaffolds. Selective fluorescent labeling, scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) analysis were employed to assess scaffold calcification in vitro. The process of CaSi(2)/PCL scaffold calcification under zero bias, during which calcium phosphate growth is significantly dependent on the structural degradation of CaSi(2) grains, has a similar mechanism as the calcium phosphate growth on bioactive glasses/ceramics. The biomineralization of these scaffolds is initiated solely by the silicide phase and can be accelerated by the degradation of the polymer matrix.

Repair of Large Articular Osteochondral Defects Using Hybrid Scaffolds and Bone Marrow-Derived Mesenchymal Stem Cells in a Rabbit Model

In order to evaluate the repair potential in large osteochondral defects on high load-bearing sites, a hybrid scaffold system was made of three-dimensional porous Polycaprolactone (PCL) scaffold for the cartilage and tricalcium phosphate-reinforced PCL for the bone portion. Osteochondral defects of 4-mm diameter x 5.5-mm depth were created in the medial femoral condyle of adult New Zealand White rabbits. The defects were treated with hybrid scaffolds without cells (control group) or seeded with allogenic bone marrow-derived mesenchymal stem cells (BMSC) in each part (experimental group) by press-fit implantation. Implanted cells were tracked using Adeno-LacZ labeling. Repair tissues were evaluated at 3 and 6 months after implantation. Overall, the experimental group showed superior repair results as compared to the control group using gross examination, qualitative and quantitative histology, and biomechanical assessment. With BMSC implantation, the hybrid scaffolds provided sufficient support to new osteochondral tissues formation. The bone regeneration was consistently good from 3 to 6 months with firm integration to the host tissue. Cartilage layer resurfacing was more complicated. All of the samples showed cartilage tissues mixed with PCL scaffold filaments at 3 months. Histology at 6 months revealed some degradation phenomenon in several samples whereas others had a good appearance; however, the Young's moduli from the experimental group (0.72 MPa) approached that of normal cartilage (0.81 MPa). In vivo viability of implanted cells was demonstrated by the retention for 6 weeks in the scaffolds. This investigation showed that PCL-based hybrid scaffolds with BMSC may be an alternative treatment for large osteochondral defects in high-loading sites.

A comparison between the classical approach and a Volterra-Wiener constitutive model in evaluating the viscoelastic properties of mouse medial collateral ligaments

The effect of melt electrospun writing fiber arrangement on cellular behavior has not yet been thoroughly investigated. Cellular orientation is particularly important in the context of ligament tissue engineering for orthopedic applications whereby a high degree of cell alignment is present in the native tissue. The aim of this study was to investigate the response of human mesenchymal stem cells (hMSC) to three different patterned porous polycaprolactone scaffolds (aligned, crimped and random) fabricated by melt electrospinning writing, resulting in 20 μm diameter electrospun fibers. Cell orientation was investigated over 4 weeks in vitro and it was demonstrated that the aligned pattern was capable of orientating the hMSCs towards the main direction of the fibers and this feature was maintained over the entire culture period whereas the orientation was rapidly lost in the crimped pattern. In order to fabricate a functional scaffold for ligament tissue engineering, the scaffolds were rolled in three bundles, subsequently braided and combined with a bone compartment (consisting of a melt electrospun scaffold seeded with osteogenically induced hMSCs) for the development of a Bone-Ligament-Bone (BLB) construct. The mechanical properties of non-cellularized and cellularized BLB constructs were assessed under both quasi-static and cyclic conditions. This revealed that the in vitro maturation significantly softened the BLB constructs and that the mechanical properties were several fold lower than those of native tissue. The cyclic testing demonstrated that the presence of cell sheets resulted in increased resilience and elasticity, even though the global mechanical properties were decreased for the in vitro matured constructs (regardless of the pattern). In conclusion, we demonstrated that melt electrospinning writing fiber organization can induce spontaneous cell alignment and that large cellularized BLB constructs with complex geometry can achieve mechanical resilience under cyclic stretching.

Fabrication of polycaprolactone scaffolds using a sacrificial compression‐molding process

A method of compression-molding fine-powder blends of polycaprolactone (PCL) and poly(ethylene oxide) (PEO) and subsequently dissolving the PEO phase was investigated to prepare porous PCL scaffolds. Different mixing ratios of the two polymers from 20 to 70% PCL were used to study the effect of the mixing ratio on the morphology formation of the scaffold. The mixing ratio was found to play an important role in affecting the porosity of the scaffold and the size of pores. Murine embryonic stem cell derived osteogenic cells were utilized to test the suitability of these scaffolds in tissue engineering applications. The seeded cells were able to colonize and grow in these scaffolds. Based on the overall consideration of morphology, mechanical performance, and ability for cell attachment and proliferation, the scaffolds with approximately 30-40% PCL appear to be an appropriate choice for tissue engineering. These findings suggest that sacrificial compression-molding of PCL-PEO powder blends can be used in the generation of biocompatible scaffolds with controllable porosity and pore size and may be used for in vitro tissue engineering applications.