Electrospun Polycaprolactone Scaffolds Research Articles

Polyvinyl alcohol-silk fibroin composite stents: A comprehensive investigation into biocompatibility and mechanical performance

Bioresorbable stents (BRS) are manufactured using biodegradable materials. As an alternative to those commonly used in commercial stents, this study explored the development of BRS using polyvinyl alcohol (PVA) and silk fibroin (SF). PVA is a promising material for the fabrication of BRS due to its biocompatibility and mechanical attributes, closely resembling those of aortic vessels. However, its application presents challenges in terms of cell adhesion and proliferation. SF has been extensively studied for its potential to enhance cell adhesion and proliferation, making it a promising biomaterial in the field of medical devices. SF was introduced by dissolving it in a PVA solution or by coating the hydrogel surface with a layer of SF. Initial tests revealed that overnight incubation of fetal bovine serum significantly increased cell viability in hydrogels. Viability assays confirmed that SF substantially improved cell viability compared to PVA alone. The method was extended to fabricate SF-coated stents, which demonstrated robust cell proliferation and improved performance compared to electrospun polycaprolactone scaffolds. In addition, the SF-coated stents displayed an increase in compressive strength, demonstrating improved biocompatibility and mechanical performance. Dynamic mechanical analysis evaluated the positive impact of SF on stent properties at physiological temperatures. The study revealed that PVA-SF stents offer a compromise between biocompatibility, mechanical strength, and elastic recovery, positioning them as a valuable alternative for cardiovascular stent applications. The dual benefits of enhanced biocompatibility and improved mechanical performance make SF-coated stents promising candidates for bioresorbable stent design.  

Sputtered zirconium based metallic glassy thin films onto electrospun PCL nanofibrous scaffolds for enriching bioactivity

Surface modification of electrospun Polycaprolactone (PCL) nanoscaffold is to enhance surface properties, particularly bioactivity. In this paper, electrospinning was employed to produce PCL nanoscaffolds and enhancing the bioactivity of the electrospun PCL scaffolds by sputtering of Zirconium based thin film metallic glasses (Zr -TFMGs). The Energy Dispersive Spectroscopy (EDS) analysis exhibited the presence of Zr, Ti and Si and surface topography by Atomic Force Microscopy (AFM) revealed the smooth surface of metallic glasses with nanofibrous structures. The sputtered samples were subjected to immersion test in Simulated Body Fluid (SBF) to analyze the apatite forming ability of Zr–Ti–Si metallic glasses. After 21days of incubation, SEM clearly showed the spherical structures of apatite formed on the surface of modified electrospun PCL. Cell viability studies using L929 fibroblast cells indicates the higher proliferation rate for the Zr based TFMG sputtered PCL scaffold specimen than the uncoated neat PCL scaffold and indicating the bioactive nature of Zr based TFMG. Contact angle of TFMG sputtered sample exhibited the hydrophobic surface which is helpful in preventing the biomedical devices from blood coagulation.

Comparative analysis of scaffold fibre diameter-modulated responses on immortalized and primary thyroid cells: Implications for thyroid regeneration

Primary hypothyroidism results from thyroid gland damage, commonly due to radiotherapy for head and neck cancer. Primary hypothyroidism severely impacts patient quality of life and is expected to increase in incidence due to the aging population and rising cancer survivor rates. Tissue engineering approaches are starting to emerge as a potential treatment.Electrospinning techniques have yet to be investigated in conjunction with thyroid tissue. Here, electrospun polycaprolactone scaffolds with varying fibre diameters were fabricated and their effects on immortalized human thyrocytes (Nthy-ori 3–1) and primary murine thyroid cells were investigated. Four groups of scaffolds were produced with fibre diameters ranging from approximately 0.5 µm to 4 µm, to understand the impact of these microenvironments on their potential for thyroid regeneration.Results showed that fibre size influenced mechanical properties, porosity and cell response. Nthy-ori 3–1 cells performed best on the scaffold with medium-sized fibres while primary cells preferred the scaffold with larger fibres. Nonetheless, the scaffold with the smallest fibre diameter encouraged the secretion of collage-like fibrils, potentially indicating favourable conditions for regeneration.This study highlights the potential of electrospun PCL scaffolds as promising candidates for tissue engineering applications in the treatment of primary hypothyroidism. The findings demonstrate the importance of tailoring scaffold properties and considering cell-type specific responses when designing biomaterials for tissue-engineered solutions for thyroid regeneration. Further research is necessary to elucidate the underlying mechanisms and optimize scaffold properties for successful thyroid tissue regeneration.

Simple Electrospinning Method for Biocompatible Polycaprolactone β-Carotene Scaffolds: Advantages and Limitations.

In this study, electrospun scaffolds were fabricated using polycaprolactone (PCL) loaded with varying concentrations of β-carotene (1.2%, 2.4%, and 3.6%) via the electrospinning technique. The electrospinning process involved the melting of PCL in acetic acid, followed by the incorporation of β-carotene powder under constant stirring. Raman spectroscopy revealed a homogeneous distribution of β-carotene within the PCL matrix. However, the β-carotene appeared in particulate form, rather than being dissolved and blended with the PCL matrix, a result also confirmed by thermogravimetric analysis. Additionally, X-ray diffraction analysis indicated a decrease in crystallinity with increasing β-carotene concentration. Mechanical testing of the scaffolds demonstrated an increase in ultimate strain, accompanied by a reduction in ultimate stress, indicating a potential plasticizing effect. Moreover, antimicrobial assays revealed a marginal antibacterial effect against Escherichia coli for scaffolds with higher β-carotene concentrations. Conversely, preliminary biological assessment using KUSA-A1 mesenchymal cells indicated enhanced cellular proliferation in response to the scaffolds, suggesting the potential biocompatibility and cell-stimulating properties of β-carotene-loaded PCL scaffolds. Overall, this study provides insights into the fabrication and characterization of electrospun PCL scaffolds containing β-carotene, laying the groundwork for further exploration in tissue engineering and regenerative medicine applications.

Evaluation of osteoconductive effect of polycaprolactone (PCL) scaffold treated with fibronectin on adipose-derived mesenchymal stem cells (AD-MSCs).

Replacing damaged organs or tissues and repairing damage by tissue engineering are attracting great interest today. A potentially effective method for bone remodeling involves combining nanofiber scaffolds with extracellular matrix (ECM), and growth factors. Today, electrospun PCL-based scaffolds are widely used for tissue engineering applications. In this study, we used an electrospun polycaprolactone (PCL) scaffold coated with fibronectin (Fn), a ubiquitous ECM glycoprotein, to investigate the induction potential of this scaffold in osteogenesis with adipose-derived mesenchymal stem cells (AD-MSCs). Scanning electron microscopy (SEM) analysis showed that fibronectin, by binding to the membrane receptors of mesenchymal stem cells (MSCs), leads to their attachment and proliferation on the PCL scaffold and provides a suitable environment for osteogenesis. In addition, biochemical tests showed that fibronectin leads to increased calcium deposition. The results also showed that alkaline phosphatase activity was significantly higher in the PCL scaffold coated with fibronectin than in the control groups (PCL scaffold group and tissue culture polystyrene (TCPS) group) (P<0.05). Also, the analysis of quantitative reverse transcription PCR (qRT-PCR) data showed that the relative expression of bone marker genes such as osteonectin (ON), osteocalcin (OC), RUNX family transcription factor 2 (RUNX2), and collagen type I alpha 1 (COL1) was much higher in the cells seeded on the PCL/Fn scaffold than in the other groups (P<0.05). The results show that fibronectin has an increasing effect in accelerating bone formation and promising potential for use in bone tissue engineering.

Bioreactor Design for Culturing Vascularized Engineered Tissue in Flow Conditions.

Background: Current treatments for congenital heart defects often require surgery and implantation of a synthetic patch or baffle that becomes a fibrous scar and leads to a high number of reoperations. Previous studies in rats have shown that a prevascularized scaffold can integrate into the heart and result in regions of vascularized and muscularized tissue. However, increasing the thickness of this scaffold for use in human hearts requires a method to populate the thick scaffold and mature it under physiologic flow and electrical conditions. Experiment: We developed a bioreactor system that can perfuse up to six 7 mm porous scaffolds with tunable gravity-mediated flow and chronic electrical stimulation. Three polymers, which have been reported to be biocompatible, were evaluated for effects on the viability of induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM). Bioreactor flow and electrical stimulation functions were tested, and the bioreactor was operated for up to 7 days to ensure reliability and lack of leaks in a 37°C, humidified incubator. Height and flow relationships were measured for perfusion through an electrospun polycaprolactone and gelatin scaffold, previously reported by our laboratory. Culture with cells was evaluated by plating human umbilical vein endothelial cells and human dermal fibroblasts on top of the scaffolds in both static and flow conditions for 2, 5, and 7 days. As a proof-of concept, scaffolds were cryosectioned and cell infiltration was quantified using immunofluorescence staining. Results: Neither MED610 (Stratasys), Vero (Stratasys), nor FORMLAB materials affected the viability of iPSC-CM, and MED610 was chosen for manufacture due to familiarity of 3D printing from this material. The generation of electrical field stimulation from 0 to 5 V and physiological ranges of pump capacities were verified. The relationship between height and flow was calculated for scaffolds with and without cells. Finally, we demonstrated evaluation of cell depth and structure in scaffolds cultured for 2, 5, and 7 days. Conclusion: The gravity-mediated flow bioreactor system we developed can be used as a platform for 3D cell culture particularly designed for perfusing vascularized tissue constructs with electrical stimulation for cardiac maturation.

Enhanced Antibacterial Ability of Electrospun PCL Scaffolds Incorporating ZnO Nanowires.

The infection of implanted biomaterial scaffolds presents a major challenge. Existing therapeutic solutions, such as antibiotic treatment and silver nanoparticle-containing scaffolds are becoming increasingly impractical because of the growth of antibiotic resistance and the toxicity of silver nanoparticles. We present here a novel concept to overcome these limitations, an electrospun polycaprolactone (PCL) scaffold functionalised with zinc oxide nanowires (ZnO NWs). This study assessed the antibacterial capabilities and biocompatibility of PCL/ZnO scaffolds. The fabricated scaffolds were characterised by SEM and EDX, which showed that the ZnO NWs were successfully incorporated and distributed in the electrospun PCL scaffolds. The antibacterial properties were investigated by co-culturing PCL/ZnO scaffolds with Staphylococcus aureus. Bacterial colonisation was reduced to 51.3% compared to a PCL-only scaffold. The biocompatibility of the PCL/ZnO scaffolds was assessed by culturing them with HaCaT cells. The PCL scaffolds exhibited no changes in cell metabolic activity with the addition of the ZnO nanowires. The antibacterial and biocompatibility properties make PCL/ZnO a good choice for implanted scaffolds, and this work lays a foundation for ZnO NWs-infused PCL scaffolds in the potential clinical application of tissue engineering.

Electrospun polycaprolactone incorporated with fluorapatite nanoparticles composite scaffolds enhance healing of experimental calvarial defect on rats.

Composite scaffolds that maximize the advantages of different polymers are widely utilized in guided tissue regeneration (GTR). Some studies found that novel composite scaffolds composed of electrospun polycaprolactone/fluorapatite (ePCL/FA) actively promoted the osteogenic mineralization of various cell types in vitro. However, only a few studies have addressed the application of this composite scaffold membrane material in vivo. In this study, the ability of ePCL/FA composite scaffolds in vivo and their possible mechanisms were preliminarily explored. In this study, ePCL/FA composite scaffolds were characterized and their effects on bone tissue engineering and repair of calvarial defects in rats were examined. Sixteen male Sprague-Dawley (SD) rats were randomly categorized into four groups: normal group (integral cranial structure without defect), control group (cranial defect), ePCL group (cranial defect repaired by electrospun polycaprolactone scaffolds), and ePCL/FA group (cranial defect repaired by fluorapatite-modified electrospun polycaprolactone scaffolds). At 1 week, 2 months, and 4 months, micro-computed tomography (micro-CT) analysis was performed to compare the bone mineral density (BMD), bone volume (BV), tissue volume (TV), and bone volume percentage (BV/TV). The effects of bone tissue engineering and repair were observed by histological examination (hematoxylin and eosin, Van Gieson, and Masson respectively) at 4 months. In water contact angle measurement, the average contact angle for the ePCL/FA group was significantly lower than that for the ePCL group, indicating that the FA crystal improved the hydrophilicity of the copolymer. Micro-CT analysis revealed that the cranial defect had no significant change at 1 week; however, the BMD, BV, and BV/TV of the ePCL/FA group were significantly higher than those of the control group at 2 and 4 months. Histological examination showed that the cranial defects were almost completely repaired by the ePCL/FA composite scaffolds at 4 months compared to the control and ePCL groups. The introduction of a biocompatible FA crystal improved the physical and biological properties of the ePCL/FA composite scaffolds; thus, these scaffolds demonstrate outstanding osteogenic potential for bone and orthopedic regenerative applications.

Reduced Fibroblast Activation on Electrospun Polycaprolactone Scaffolds.

In vivo, quiescent fibroblasts reside in three-dimensional connective tissues and are activated in response to tissue injury before proliferating rapidly and becoming migratory and contractile myofibroblasts. When deregulated, chronic activation drives fibrotic disease. Fibroblasts cultured on stiff 2D surfaces display a partially activated phenotype, whilst many 3D environments limit fibroblast activation. Cell mechanotransduction, spreading, polarity, and integrin expression are controlled by material mechanical properties and micro-architecture. Between 3D culture systems, these features are highly variable, and the challenge of controlling individual properties without altering others has led to an inconsistent picture of fibroblast behaviour. Electrospinning offers greater control of mechanical properties and microarchitecture making it a valuable model to study fibroblast activation behaviour in vitro. Here, we present a comprehensive characterisation of the activation traits of human oral fibroblasts grown on a microfibrous scaffold composed of electrospun polycaprolactone. After over 7 days in the culture, we observed a reduction in proliferation rates compared to cells cultured in 2D, with low KI67 expression and no evidence of cellular senescence. A-SMA mRNA levels fell, and the expression of ECM protein-coding genes also decreased. Electrospun fibrous scaffolds, therefore, represent a tuneable platform to investigate the mechanisms of fibroblast activation and their roles in fibrotic disease.

Erratum to "Controlled Vacuum Seeding as a Means of Generating Uniform Cellular Distribution in Electrospun Polycaprolactone (PCL) Scaffolds," ASME J. Biomech. Eng., 2009, 131(7), p. 074521, DOI: 10.1115/1.3173283.

Erratum of figures and tables

Activated carbon nanofiber nanoparticles incorporated electrospun polycaprolactone scaffolds to promote fibroblast behaviors for application to skin tissue engineering

The most widely used one-dimensional (1D) carbonaceous nanomaterials in tissue engineering are carbon nanotubes, either single or multiwalled. Other forms of 1D nanomaterials, such as carbon nanowires and carbon nanofibers, have been less explored for biomedical applications. Herein, we synthesized 1D-activated carbon nanofiber nanoparticles (ACNF NPs) from the polyacrylonitrile electrospun nanofibers by continuous processes like stabilization, alkali treatment, calcination, and grinding. Two different sets of ACNF NPs-containing electrospun polycaprolactone (PCL) nanofiber mats, viz. surface-modified NP-deposited mats (ACNF@PCL) and NP-incorporated mats (ACNF-PCL), were prepared to examine their potential as skin tissue engineering scaffolds. Raman spectra demonstrated that ACNF NPs exhibited graphitization degree with an ID/IG ratio of 1.05. Scanning electron microscopy (SEM) observations showed that ACNF NPs are sized 280 ± 100 nm by diameter and 565–3322 nm by length. The NPs concentrated above 30 μg/mL were found to exhibit toxicity with < 70% viability of NIH3T3 fibroblasts on 48 h. The ACNF-PCL nanofiber mats displayed better cell proliferation profile showing significant changes compared to PCL and ACNF@PCL mats on days 1, 3, and 5. Hence, we concluded that ACNF-PCL mats with less concentration of ACNF NPs have more potential to support cellular growth, ensuring its possible impact on skin tissue regeneration.Graphical Abstract

Hybrid 3D Printed and Electrospun Multi-Scale Hierarchical Polycaprolactone Scaffolds to Induce Bone Differentiation.

Complex scaffolds composed of micro- and nano-structures are a key target in tissue engineering and the combination of sequential 3D printing and electrospinning enables the fabrication of these multi-scale structures. In this work, dual 3D printed and electrospun polycaprolactone (PCL) scaffolds with multiple mesh layers were successfully prepared. The scaffold macro- and micro-porosity were assessed by optical and scanning electron microscopy, showing that electrospun fibers formed aligned meshes within the pores of the scaffold. Consequently, the hydrophilicity of the scaffold increased with time, enhancing cell adhesion and growth. Additionally, compression tests in back and forth cycles demonstrated a good shape recovery behavior of the scaffolds. Biological results indicated that hybrid PCL scaffolds are biocompatible and enable a correct cell culture over time. Moreover, MC3T3-E1 preosteoblast culture on the scaffolds promoted the mineralization, increased the alkaline phosphatase (ALP) activity and upregulated the expression of early and late osteogenic markers, namely ALP and osteopontin (OPN), respectively. These results demonstrate that the sequential combination of 3D printing and electrospinning provides a facile method of incorporating fibers within a 3D printed scaffold, becoming a promising approach towards multi-scale hierarchical scaffolds capable of guiding the osteogenic differentiation.

A novel approach to bio-mineralization of electrospun PCL scaffolds by protein and hydroxyapatite nanoparticles; molecular dynamics simulation and in-vitro evaluation

The nano-fibrous polycaprolactone (PCL) scaffolds are one of the promising materials for bone tissue engineering. However, the role of protein in the bio-mineralization of PCL fibers is not clear. This study aims to find how a protein (e.g. bovine serum albumin (BSA)) is attached to the hydroxyapatite (HA) nanoparticles (NPs) loaded on the PCL fiber, and it affected the bio-mineralization of a hybrid scaffold. The molecular dynamic (MD) simulation was utilized to explore the protein and HA attachment. The results of MD simulation indicated that the a(b)-planes of HA crystal and glutamic acid (Glu) from BSA molecule have critical roles in attachment together. In the following, the electrospinning method was applied to synthesize the pristine and hybrid PCL scaffolds to evaluate the MD results. The scanning electron microscopy (SEM) images showed that BSA could be successfully attached to the HA NPs, and coiled around the PCL-HA junction confirming the MD simulation. To evaluate the bio-mineralization, the in-vitro immersion test in the simulation body fluid (SBF) solution and the cytocompatibility of scaffolds by the MG63 osteosarcoma cells were studied. The results indicated better osteoconduction and bone regeneration for the hybrid PCL- BSA-HA scaffold because of the synergic effects of HA NPs and BSA protein.

Fabrication and Characterization of Decellularized Periodontal Ligament Cell Sheet Constructs.

Decellularized tissue engineered constructs have the potential to promote regeneration by providing a biomimetic extracellular matrix that directs tissue specific regeneration when implanted in situ. Recently, the use of cell sheets has shown promising results in promoting periodontal regeneration. Here, we describe the fabrication of decellularized periodontal cell sheets with intact extracellular matrix structural and biological properties. Melt electro-spun polycaprolactone (PCL) scaffolds are used as a carrier for the inherently fragile cell sheets, in order to provide support during the processes of decellularization. An optimized decellularization method is outlined using perfusion with a combination of NH4OH and Triton X-100 together with a DNase treatment step for DNA removal. The maintenance of extracellular matrix structural and biological integrity is important, and here, we describe the assessment of these properties using immunostaining for extracellular matrix proteins and ELISA for growth factor quantification.

The solvent chosen for the manufacturing of electrospun polycaprolactone scaffolds influences cell behavior of lung cancer cells

The development of a trustworthy in vitro lung cancer model is essential to better understand the illness, find novel biomarkers, and establish new treatments. Polycaprolactone (PCL) electrospun nanofibers are a cost-effective and ECM-like approach for 3D cell culture. However, the solvent used to prepare the polymer solution has a significant impact on the fiber morphology and, consequently, on the cell behavior. Hence, the present study evaluated the effect of the solvent employed in the manufacturing on the physical properties of 15%-PCL electrospun scaffolds and consequently, on cell behavior of NCI-H1975 lung adenocarcinoma cells. Five solvents mixtures (acetic acid, acetic acid-formic acid (3:1, v/v), acetone, chloroform-ethanol (7:3, v/v), and chloroform-dichloromethane (7:3, v/v)) were tested. The highest cell viability (overline{x } = 33.4%) was found for cells cultured on chloroform-ethanol (7:3) PCL scaffolds. Chloroform-dichloromethane (7:3) PCL scaffolds exhibited a roughness that enhanced the quality of electrospun filament, in terms of cell viability. Our findings highlighted the influence of the solvent on fiber morphology and protein adsorption capacity of nanofilaments. Consequently, these features directly affected cell attachment, morphology, and viability.

Three-dimensional wet electrospun scaffold system for the differentiation of adipose-derived mesenchymal stem cells to islet-like clusters.

Stem cell-derived islet-like clusters (ILCs) are an alternative source of pancreatic beta cells for the treatment of diabetic mellitus. An ideal 3D culture platform for the generation of ILCs of desired cluster size is a challenge due to the clustering of islet cells in the 2D culture systems. The islet cells cultured in 2D conditions produce clusters of large size, which are less efficient in terms of insulin secretion and viability. In this study, we report that ILCs formed on a PCL-based wet electrospun fibrous scaffold with larger pore size produced clusters of the desired size, compared to that cultured on a conventional electrospun sheet. The collagen functionalization on this wet electrospun polycaprolactone (PCL) scaffold showed enhanced insulin secretion and cell viability compared to the non-functionalized or conventionally electrospun PCL scaffold. The collagen-coated wet electrospun 3D scaffold produced ILCs of cluster diameter 70±20μm and the conventionally electrospun PCL sheet produced larger ILC clusters of diameter 300±10μm. Hence the results indicate the collagen-functionalized wet electrospun scaffold system could be a potential scaffold for islet tissue engineering.

Electrospun polycaprolactone scaffolds loaded with a 1,4-naphthoquinone derivative for anticancer therapy

• Novel materials for tissue repair after surgical resection of tumors are needed. • Naphthoquinones have been reported to be effective for anticancer therapy. • Electrospun PCL-based scaffolds were loaded with a 1,4-naphthoquinone derivative. • Anticancer activity was retained, yet mechanical properties did not change for doped scaffolds. The development of bioresorbable scaffolds with anticancer properties for use in tissue repair after surgical resection of tumors is important. Naphthoquinones with high anticancer activity are promising compounds for creating anticancer scaffolds. Thus, electrospun polycaprolactone (PCL) scaffolds embedded with different concentrations of 2-chloro-3-((4-hydroxyphenyl)amino)-1,4-naphthoquinonone (NQCP4) were fabricated without changing the technological parameters of scaffold formation. Loading NQCP4 into the spinning solution at concentrations of 10 to 50 μg/mL did not cause changes in the structure, strength, or wettability of the scaffold. However, increasing NQCP4 concentrations in the spinning solution enhanced anticancer properties of the scaffolds in vitro . Thus, the use of NQCP4 makes it possible to obtain promising PCL scaffolds for tissue repair, which are in demand for surgical resection of malignant neoplasms, while still maintaining the technological advantages of “classical” electrospinning technology.

Effect of addition of Layered Double Hydroxides (LDH) on mechanical and biological properties of electrospun polycaprolactone scaffold

Effect of addition of Layered Double Hydroxides (LDH) on mechanical and biological properties of electrospun polycaprolactone scaffold

Rat liver ECM incorporated into electrospun polycaprolactone scaffolds as a platform for hepatocyte culture.

Liver disease is expanding across the globe; however, health-care systems still lack approved pharmaceutical treatment strategies to mitigate potential liver failures. Organ transplantation is the only treatment for liver failure and with increasing cases of liver disease, transplant programs increasingly cannot provide timely transplant availability for all patients. The development of pharmaceutical mitigation strategies is clearly necessary and methods to improve drug development processes are considered vital for this purpose. Herein, we present a methodology for incorporating whole organ decellularised rat liver ECM (rLECM) into polycaprolactone (PCL) electrospun scaffolds with the aim of producing biologically relevant liver tissue models. rLECM PCL scaffolds have been produced with 5w/w% and 10w/w% rLECM:PCL and were analyzed by SEM imaging, tensile mechanical analyses and FTIR spectroscopy. The hepatocellular carcinoma cell line, HepG2, was cultured upon the scaffolds for 14 days and were analyzed through cell viability assay, DNA quantification, albumin quantification, immunohistochemistry, and RT-qPCR gene expression analysis. Results showed significant increases in proliferative activity of HepG2 on rLECM containing scaffolds alongside maintained key gene expression. This study confirms that rLECM can be utilized to modulate the bioactivity of electrospun PCL scaffolds and has the potential to produce electrospun scaffolds suitable for enhanced hepatocyte cultures and in-vitro liver tissue models.

Functionalized Electrospun Scaffold-Human-Muscle-Derived Stem Cell Construct Promotes In Vivo Neocartilage Formation.

Polycaprolactone (PCL) is a non-cytotoxic, completely biodegradable biomaterial, ideal for cartilage tissue engineering. Despite drawbacks such as low hydrophilicity and lack of functional groups necessary for incorporating growth factors, it provides a proper environment for different cells, including stem cells. In our study, we aimed to improve properties of scaffolds for better cell adherence and cartilage regeneration. Thus, electrospun PCL–scaffolds were functionalized with ozone and loaded with TGF-β3. Together, human-muscle-derived stem cells (hMDSCs) were isolated and assessed for their phenotype and potential to differentiate into specific lineages. Then, hMDSCs were seeded on ozonated (O) and non-ozonated (“naïve” (NO)) scaffolds with or without protein and submitted for in vitro and in vivo experiments. In vitro studies showed that hMDSC and control cells (human chondrocyte) could be tracked for at least 14 days. We observed better proliferation of hMDSCs in O scaffolds compared to NO scaffolds from day 7 to day 28. Protein analysis revealed slightly higher expression of type II collagen (Coll2) on O scaffolds compared to NO on days 21 and 28. We detected more pronounced formation of glycosaminoglycans in the O scaffolds containing TGF-β3 and hMDSC compared to NO and scaffolds without TGF-β3 in in vivo animal experiments. Coll2-positive extracellular matrix was observed within O and NO scaffolds containing TGF-β3 and hMDSC for up to 8 weeks after implantation. These findings suggest that ozone-treated, TGF-β3-loaded scaffold with hMDSC is a promising tool in neocartilage formation.