Polycaprolactone Nanofibrous Scaffolds Research Articles

Soy Protein Nanofiber Scaffolds for Uniform Maturation of Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium.

Retinal pigment epithelium (RPE) differentiated from human induced pluripotent stem cells, called induced retinal pigment epithelium (iRPE), is being explored as a cell-based therapy for the treatment of retinal degenerative diseases, especially age-related macular degeneration. The success of RPE implantation is linked to the use of biomimetic scaffolds that simulate Bruch's membrane and promote RPE maturation and integration as a functional tissue. Due to difficulties associated with animal protein-derived scaffolds, including sterility and pro-inflammatory responses, current practices favor the use of synthetic polymers, such as polycaprolactone (PCL), for generating nanofibrous scaffolds. In this study, we tested the hypothesis that plant protein-derived fibrous scaffolds can provide favorable conditions permissive for the maturation of RPE tissue sheets in vitro. Our natural, soy protein-derived nanofibrous scaffolds exhibited a J-shaped stress-strain curve that more closely resembled the mechanical properties of native tissues than PCL with significantly higher hydrophilicity of the natural scaffolds, favoring in vivo implantation. We then demonstrate that iRPE sheets growing on these soy protein scaffolds are equivalent to iRPE monolayers cultured on synthetic PCL nanofibrous scaffolds. Immunohistochemistry demonstrated RPE-like morphology and functionality with appropriate localization of RPE markers RPE65, PMEL17, Ezrin, and ZO1 and with anticipated histotypic polarization of vascular endothelial growth factor and pigment epithelium-derived growth factor as indicated by enzyme-linked immunosorbent assay. Scanning electron microscopy revealed dense microvilli on the cell surface and homogeneous tight junctional contacts between the cells. Finally, comparative transcriptome analysis in conjunction with principal component analysis demonstrated that iRPE on nanofibrous scaffolds, either natural or synthetic, matured more consistently than on nonfibrous substrates. Taken together, our studies suggest that the maturation of cultured iRPE sheets for subsequent clinical applications might benefit from the use of nanofibrous scaffolds generated from natural proteins. Impact statement Induced retinal pigment epithelium (iRPE) from patient-derived induced pluripotent stem cells (iPSCs) may yield powerful treatments of retinal diseases, including age-related macular degeneration. Recent studies, including early human clinical trials, demonstrate the importance of selecting appropriate biomaterial scaffolds to support tissue-engineered iRPE sheets during implantation. Electrospun scaffolds show particular promise due to their similarity to the structure of the native Bruch's membrane. In this study, we describe the use of electroprocessed nanofibrous soy protein scaffolds to generate polarized sheets of human iPSC-derived iRPE sheets. Our evaluation, including RNA-seq transcriptomics, indicates that these scaffolds are viable alternatives to scaffolds electrospun from synthetic polymers.

Preparation of electrospun nanofibrous polycaprolactone scaffolds using nontoxic ethylene carbonate and glacial acetic acid solvent system

ABSTRACTIn recent years, solution electrospinning has attracted the interest of researchers due to the possibility to design nanofibrous scaffolds with large surface area to volume ratios. Polycaprolactone (PCL), because of its biocompatibility and easy processability, has been widely used to develop electrospun structures for tissue engineering. However, the use of organic solvents and the poor PCL solution stability still hinder the development of the solution electrospinning process. The relatively benign glacial acetic acid (GAC) as a solvent of PCL was used to fabricate microfibrous fibers or beaded fibers. Thus, ethylene carbonate (EC) as a nontoxic assistant solvent was added to the PCL/GAC solution to successfully fabricate electrospun nanofibrous PCL scaffolds. The stability of the PCL/GAC/EC solution system was demonstrated as the viscosity, which showed no significant change during 48 h. The ultrafine PCL fiber diameter decreased as EC concentration was increased from 0 to 9 vol% and started to slightly increase when EC concentration increased beyond 9 vol%. MTT assay evidenced that MC3T3‐E1 cells on the nanofibrous PCL scaffolds exhibited a better enhancement on cell proliferation. In summary, EC was added in PCL/GAC to establish a stable and low toxic solution electrospinning system, which provides promising strategy in tissue engineering field. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020, 137, 48387.

Plasma-Coated Polycaprolactone Nanofibers with Covalently Bonded Platelet-Rich Plasma Enhance Adhesion and Growth of Human Fibroblasts.

Biodegradable nanofibers are extensively employed in different areas of biology and medicine, particularly in tissue engineering. The electrospun polycaprolactone (PCL) nanofibers are attracting growing interest due to their good mechanical properties and a low-cost structure similar to the extracellular matrix. However, the unmodified PCL nanofibers exhibit an inert surface, hindering cell adhesion and negatively affecting their further fate. The employment of PCL nanofibrous scaffolds for wound healing requires a certain modification of the PCL surface. In this work, the morphology of PCL nanofibers is optimized by the careful tuning of electrospinning parameters. It is shown that the modification of the PCL nanofibers with the COOH plasma polymers and the subsequent binding of NH2 groups of protein molecules is a rather simple and technologically accessible procedure allowing the adhesion, early spreading, and growth of human fibroblasts to be boosted. The behavior of fibroblasts on the modified PCL surface was found to be very different when compared to the previously studied cultivation of mesenchymal stem cells on the PCL nanofibrous meshes. It is demonstrated by X-ray photoelectron spectroscopy (XPS) that the freeze–thawed platelet-rich plasma (PRP) immobilization can be performed via covalent and non-covalent bonding and that it does not affect biological activity. The covalently bound components of PRP considerably reduce the fibroblast apoptosis and increase the cell proliferation in comparison to the unmodified PCL nanofibers or the PCL nanofibers with non-covalent bonding of PRP. The reported research findings reveal the potential of PCL matrices for application in tissue engineering, while the plasma modification with COOH groups and their subsequent covalent binding with proteins expand this potential even further. The use of such matrices with covalently immobilized PRP for wound healing leads to prolonged biological activity of the immobilized molecules and protects these biomolecules from the aggressive media of the wound.

Adipose-derived stem cells-conditioned medium improved osteogenic differentiation of induced pluripotent stem cells when grown on polycaprolactone nanofibers.

Considering that the common osteogenic growth factors cannot be transplanted with stem cells to the patients, many studies are underway to find a replacement for these factors. Recently, it has been determined that mesenchymal stem cell (MSC)-derived conditioned medium (CM) contains effective factors in the bone formation process. In the current study, the synergistic effect of adipose-derived MSC's CM, and polycaprolactone (PCL) scaffold was investigated on the osteogenic differentiation potential of human induced pluripotent stem cells (iPSCs). After scaffold fabrication by electrospinning and characterization by scanning electron microscopy, iPSCs proliferation in the presence of CM, PCL, and both was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide. Then, iPSCs osteogenic differentiation was investigated while cultured on tissue culture plate and PCL under CM compared with the osteogenic medium using alizarin red staining, calcium content, alkaline phosphatase activity and gene and protein expression analysis. Proliferation rate of the iPSCs was increased while cultured under CM and its effect was synergistically enhanced by culture on PCL. Evaluation of the osteogenic markers was showed CM alone could induce osteogenic differentiation into the iPSCs and this potential was significantly increased while combined with PCL nanofibrous scaffold. According to the results, it was demonstrated that CM has an osteogenic induction property almost the same of the common osteogenic medium and it can also be used potentially with stem cells when transplant to the patients. CM can also help by prolonging cell survival at the site of the defect as well as accelerating healing process.

Organotypic 3D Culture in Nanoscaffold Microwells Supports Salivary Gland Stem-Cell-Based Organization.

The self-organizing properties of stem cells have been exploited to generate organoids, organ-specific, cell-containing, three-dimensional (3D) structures. The present study aimed to introduce a novel bioengineering technique for driving the effective organization of adult tissue stem cells via niche-independent 3D microwell culture. Microwells were fabricated by photopatterning poly(ethylene glycol) hydrogel in the presence of an electrospun polycaprolactone nanofibrous scaffold. Human single clonal salivary gland stem cells (SGSCs) were cultured in nanofibrous microwells through two simple steps, priming and differentiation. Before the induction of 3D organization, single clonal SGSCs were preconditioned to aggregate to form 3D spheroids in different matrices, such as Matrigel, floating dish, and microwells. Expression of salivary stem cell markers and pluripotency markers was greater in 3D spheroid cultures than in 2D plastic culture. Lobular structures were organized by changing media, and those in microwells exhibited higher salivary acinar, ductal, and tight junction marker levels and decreased stem-cell marker levels relative to other 3D cultures. Furthermore, higher α-amylase secretion and intracellular calcium levels were observed in the presence of adrenergic or cholinergic agonists, respectively, along with an increased intracellular pH, suggesting more functional salivary organoid formation. These microwell-driven organoids also engrafted successfully into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. Our results showed that microwell-cultured SGSCs organize into salivary structures and that this biomimetic 3D culture technique can promote effective generation of niche-independent single stem-cell-based 3D organoids.

Cellular interactions with bacterial cellulose: Polycaprolactone nanofibrous scaffolds produced by a portable electrohydrodynamic gun for point-of-need wound dressing.

Electrospun nanofibrous scaffolds are promising regenerative wound dressing options but have yet to be widely used in practice. The challenge is that nanofibre productions rely on bench-top apparatuses, and the delicate product integrity is hard to preserve before reaching the point of need. Timing is critically important to wound healing. The purpose of this investigation is to produce novel nanofibrous scaffolds using a portable, hand-held "gun", which enables production at the wound site in a time-dependent fashion, thereby preserving product integrity. We select bacterial cellulose, a natural hydrophilic biopolymer, and polycaprolactone, a synthetic hydrophobic polymer, to generate composite nanofibres that can tune the scaffold hydrophilicity, which strongly affects cell proliferation. Composite scaffolds made of 8 different ratios of bacterial cellulose and polycaprolactone were successfully electrospun. The morphological features and cell-scaffold interactions were analysed using scanning electron microscopy. The biocompatibility was studied using Saos-2 cell viability test. The scaffolds were found to show good biocompatibility and allow different proliferation rates that varied with the composition of the scaffolds. A nanofibrous dressing that can be accurately moulded and standardised via the portable technique is advantageous for wound healing in practicality and in its consistency through mass production.

In vitro degradation and in vivo toxicity of NanoMatrix3D® polycaprolactone and poly(lactic acid) nanofibrous scaffolds.

Nanofibrous materials present unique properties favorable in many biomedicine and industrial applications. In this research we evaluated biodegradation, tissue response and general toxicity of nanofibrous poly(lactic acid) (PLA) and polycaprolactone (PCL) scaffolds produced by conventional method of electrospinning and using NanoMatrix3D® (NM3D® ) technology. Mass density, scanning electron microscopy and in vitro degradation (static and dynamic) were used for material characterization, and subcutaneous, intramuscular and intraperitoneal implantation - for in vivo tests. Biochemical blood analysis and histology were used to assess toxicity and tissue response. Pore size and fiber diameter did not differ in conventional and NM3D® PLA and PCL materials, but mass density was significantly lower in NM3D® ones. Scaffolds made by conventional method showed toxic effect during the in-vivo tests due to residual concentration of chloroform that released with material degradation. NM3D® method allowed cleaning scaffolds from residual solutions that made them nontoxic and biocompatible. Subcutaneous, intramuscular and intraperitoneal implantation of PCL and PLA NM3D® electrospun nanofibrous scaffolds showed their appropriate cell conductive properties, tissue and vessels formation in all sites. Thus, NM3D® PCL and PLA nanofibrous electrospun scaffolds can be used in the field of tissue engineering, surgery, wound healing, drug delivery, and so forth, due to their unique properties, nontoxicity and biocompatibility. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2200-2212, 2018.

Fabrication, characterization, and biocompatibility assessment of a novel elastomeric nanofibrous scaffold: A potential scaffold for soft tissue engineering.

With regard to flexibility and strength properties requirements of soft biological tissue, elastomeric materials could be more beneficial in soft tissue engineering applications. The present work investigates the use of an elastic polymer, (polycaprolactone fumarate [PCLF]), for fabricating an electrospun scaffold. PCLF with number-average molecular weight of 13,284 g/mol was synthetized, electrospun PCLF:polycaprolactone (PCL) (70:30) nanofibrous scaffolds were fabricated and a novel strategy (in situ photo-crosslinking along with wet electrospinning) was applied for crosslinking of PCLF in the structure of PCLF:PCL nanofibers was presented. Sol fraction results, Fourier-transform infrared spectroscopy, and mechanical tests confirmed occurrence of crosslinking reaction. Strain at break and Young's modulus of crosslinked PCLF:PCL nanofibers fabricated was found to be 114.5 ± 3.9% and 0.6 ± 0.1 MPa, respectively, and dynamic mechanical analysis results revealed elasticity of nanofibers. MTS assay showed biocompatibility of PCLF:PCL (70:30) nanofibrous scaffolds. Our overall results showed that electrospun PCLF:PCL nanofibrous scaffold could be considered as a candidate for further in vitro and in vivo experiments and its application for engineering of soft tissues subjected to in vivo cyclic mechanical stresses. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2371-2383, 2018.

Evaluation of electrospinning parameters on the tensile strength and suture retention strength of polycaprolactone nanofibrous scaffolds through surface response methodology

Scaffolds should provide sufficient biomechanical support during tissue regeneration for tissue engineering (TE) applications. Electrospun scaffolds are commonly applied in TE applications due to their tunable physical, chemical, and mechanical properties as well as their similarity to extracellular matrix. Although the mechanical properties of electrospun scaffolds are highly dependent on processing parameters, a limited number of studies have systematically investigated this subject. The present study has investigated the effects of the main electrospinning parameters on tensile and suture retention strength of polycaprolactone (PCL) scaffolds using response surface methodology. Scaffolds morphology and cell-scaffold interaction were also investigated in this study. According to the fitted model, polymer concentration and feed rate have the most significant positive effect on both the tensile and suture retention strength. Whereas applied voltage negatively affected both the tensile and suture retention strength. The effect of distance on tensile strength was not significant while its effect on suture retention was different depending on its values. Changes in biomechanical properties were associated with gross alterations in morphology of the fibers and cell-scaffold interaction. Scaffolds with lowest tensile strength presented a beaded morphology while scaffolds with higher tensile strength presented beadless morphology with worm-like fibers. The increase in tensile strength was correlated with the increase in average diameter of the fibers and pore size. The results of cell culture study showed that fibroblasts stretched and proliferated more on scaffolds with lower tensile strength. The generated model might be helpful when PCL scaffold with desirable tensile and suture retention strength are required. Furthermore, the results suggest that changes in morphology and subsequent cell-scaffold interaction should be considered when these biomechanical properties are optimized.

Osteogenic differentiation of 3D cultured mesenchymal stem cells induced by bioactive peptides.

Bioactive peptides derived from receptor binding motifs of native proteins are a potent source of bioactive molecules that can induce signalling pathways. These peptides could substitute for osteogenesis promoting supplements. The work presented here compares three kinds of bioactive peptides derived from collagen III, bone morphogenetic protein 7 (BMP-7) and BMP-2 with their potential osteogenic activity on the model of porcine mesenchymal stem cells (pMSCs). pMSCs were cultured on electrospun polycaprolactone nanofibrous scaffolds with different concentrations of the bioactive peptides without addition of any osteogenic supplement. Analysis of pMSCs cultures included measurement of the metabolic activity and proliferation, immunofluorescence staining and also qPCR. Results showed no detrimental effect of the bioactive peptides to cultured pMSCs. Based on qPCR analysis, the bioactive peptides are specific for osteogenic differentiation with no detectable expression of collagen II. Our results further indicate that peptide derived from BMP-2 protein promoted the expression of mRNA for osteocalcin (OCN) and collagen I significantly compared to control groups and also supported deposition of OCN as observed by immunostaining method. The data suggest that bioactive peptide with an amino acid sequence of KIPKASSVPTELSAISTLYL derived from BMP-2 protein was the most potent for triggering osteogenic differentiation of pMSCs.

COMPARISON ON IN VITRO DEGRADATION OF POLYCAPROLACTONE AND POLYCAPROLACTONE/GELATIN NANOFIBROUS SCAFFOLD

Tissue engineering has emerged to provide a new medical therapy in helping tissue regrowth and regeneration by employing scaffold as an artificial supporting structure for cellular growth. Many polymers have been utilized in the fabrication of these artificial scaffolds, but there is still a need to fabricate hydrophilic nanofibrous scaffold with appropriate degradation rate. In this study, polycaprolactone (PCL) and polycaprolactone/gelatin (PCL/Ge) 70:30 nanofibrous scaffolds were fabricated using electrospinning technique and compared on in vitro degradation rate to determine a more suitable scaffold for skin tissue engineering application. In vitro degradation was evaluated by morphological changes, water uptake, and chemical bonding until 12 weeks. Result shows that both PCL and PCL/Ge (70:30) nanofibrous scaffolds were degraded after 8th week. However, the degradation rate of PCL nanofibrous scaffold is slower and does not has obvious morphological changes. PCL/Ge (70:30) nanofibrous scaffold with faster degradation rate have the potential for skin tissue engineering application.

Renal Differentiation of Mesenchymal Stem Cells Seeded on Nanofibrous Scaffolds Improved by Human Renal Tubular Cell Lines-Conditioned Medium.

Kidney injuries and renal dysfunctions are one of the most important clinical problems, and tissue engineering could be a valuable method for solving it. The objective of this study was to investigate the synergistic effect of renal cell line-conditioned medium and Polycaprolactone (PCL) nanofibers on renal differentiation of human mesenchymal stem cells (MSCs). In the current study, after stem cells isolation and characterization, PCL nanofibrous scaffold was fabricated using electrospinning methods and characterized morphologically, mechanically, and for biocompatibility. The renal differentiation of seeded MSCs on the surface of PCL nanofibers with and without human renal tubular cell lines-conditioned medium was investigated by evaluation of eight important renal-related genes expression by real-time reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry. Fabricated nanofibrous scaffolds were good in all characterized items. Almost highest expression of all genes was detected in stem cells seeded on PCL under conditioned media in comparison with the stem cells seeded on PCL, tissue culture polystyrene (TCPS) under renal induction medium, and TCPS under conditioned medium. According to the results, PCL nanofibers in contribution with conditioned medium can provide the optimal conditions for renal differentiation of MSCs and could be a promising candidate for renal tissue engineering application.

Development and characterization of heparin-immobilized polycaprolactone nanofibrous scaffolds for tissue engineering using gamma-irradiation

Polycaprolactone (PCL) has been considered a useful material for orthopedic devices and osseous implants because of its biocompatibility and bone-forming activity.

Effect of the dissolution time into an acid hydrolytic solvent to tailor electrospun nanofibrous polycaprolactone scaffolds

The hydrolysis of the polycaprolactone (PCL) as a function of the dissolution time in a formic/acetic acid mixture was considered as a method for tailoring the morphology of nanofibrous PCL scaffolds. Hence, the aim of this research was to establish a correlation between the dissolution time of the polymer in the acid solvent with the physico-chemical properties of the electrospun nanofibrous scaffolds and their further service life behaviour.The physico-chemical properties of the scaffolds were assessed in terms of fibre morphology, molar mass and thermal behaviour. A reduction of the molar mass and the lamellar thickness as well as an increase of the crystallinity degree were observed as a function of dissolution time. Bead-free fibres were found after 24 and 48h of dissolution time, with similar diameter distributions. The decrease of the fibre diameter distributions along with the apparition of beads was especially significant for scaffolds prepared after 72h and 120h of dissolution time in the acid mixture.The service life of the obtained devices was evaluated by means of in vitro validation under abiotic physiological conditions. All the scaffolds maintained the nanofibrous structure after 100days of immersion in water and PBS. The molar mass was barely affected and the crystallinity degree and the lamellar thickness increased along immersion, preventing scaffolds from degradation. Scaffolds prepared after 24h and 48h kept their fibre diameters, whereas those prepared after 72h and 120h showed a significant reduction.This PCL tailoring procedure to obtain scaffolds that maintain the nanoscaled structure after such long in vitro evaluation will bring new opportunities in the design of long-term biomedical patches.

Functional spheroid organization of human salivary gland cells cultured on hydrogel-micropatterned nanofibrous microwells

Three dimensional (3D) cultures of salivary glandular epithelial cells using nanofibrous bottom facilitate the formation of acinar-like organoids. In this study, we adapted a PEG hydrogel-micropatterned PCL nanofibrous microwell for the efficient bioengineering of human salivary gland organoids, in which we could easily produce uniform size of 3D organoids. This 3D culture system supports spherical organization, gene and protein expression of acinar markers, TJ proteins, adherence, and cytoskeletal proteins, as well as to promote epithelial structural integrity and acinar secretory functions, and results showed superior efficiency relative to Matrigel and nanofibrous scaffold culture. This 3D culture system has benefits in terms of inert, non-animal and serum-free culture conditions, as well as controllable spheroid size and scalable production of functional SG organoids and is applicable to bioengineering approaches for an artificial bio-gland, as well as to investigations of salivary gland physiology and regeneration.

Mesenchymal stem cells growth and proliferation enhancement using PLA vs PCL based nanofibrous scaffolds

Electrospinning of polymers is the most commonly used technique for nanofiber fabrication. polylactic acid (PLA) and polycaprolactone (PCL) have been shown to be ideal for nanofiber preparation in various biomedical applications, due to characteristics such as biodegradablity and their ability to promote the cell growth, similar to native tissues. The aim of this study was to develop biocompatible and biodegradable PLA and PCL-based nanofibrous scaffolds for enhancing stem cell growth and proliferation. The scaffolds were prepared by electrospinning, and their physicochemical properties were studied using Fourier Transform Infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and X-ray diffraction (XRD). The surface morphology of the developed scaffolds was determined using scanning electron microscopy (SEM). Mesenchymal stem cells (MSCs), derived from both adipose tissue and bone marrow, were seeded onto the prepared nanofibrous scaffolds. The effect of scaffold type, and structural characteristics on survival and proliferation of MSCs were evaluated. Our results show that after full physicochemical characterization of PCL and PLA nanofibrous scaffolds both were safe and non-toxic to the evaluated cells and both scaffolds supported cell attachment and proliferation of bone marrow and adipose tissue-derived MSCs.

Nanofibrous polycaprolactone scaffolds with adhered platelets stimulate proliferation of skin cells.

Faulty wound healing is a global healthcare problem. Chronic wounds are generally characterized by a reduction in availability of growth factors. New strategies are being developed to deliver growth factors more effectively. In this study, we introduced electrospun scaffolds composed of polycaprolactone (PCL) nanofibers functionalized with adhered platelets, as a source of numerous growth factors. Three concentrations of platelets were immobilized to nanofibrous scaffolds by simple adhesion, and their influence on adhesion, proliferation and metabolic activity of seeded cells (murine fibroblasts, keratinocytes and melanocytes) was investigated. The data obtained indicated that presence of platelets significantly promoted cell spreading, proliferation and metabolic activity in all the skin-associated cell types. There were no significant differences among tested concentrations of platelets, thus even the lowest concentration sufficiently promoted proliferation of the seeded cells. Such complex stimulation is needed for improved healing of chronic wounds. However, the nanofibrous system can be used not only as a skin cover, but also in broader applications in regenerative medicine.

Human unrestricted somatic stem cells loaded in nanofibrous PCL scaffold and their healing effect on skin defects

Unrestricted somatic stem cells (USSCs) loaded in nanofibrous polycaprolactone (PCL) scaffolds can be used for skin regeneration when grafted onto full-thickness skin defects of rats. Nanofibrous PCL scaffolds were designed by the electrospinning method and crosslinked with laminin protein. Afterwards, the scaffolds were evaluated by scanning electron microscopy, and physical and mechanical assays. In this study, nanofibrous PCL scaffolds loaded with USSCs were grafted onto the skin defects. The wounds were subsequently investigated 21 days after grafting. Results of mechanical and physical analyses showed good resilience and compliance to movement as a skin graft. In animal models; study samples exhibited the most pronounced effect on wound closure, with statistically significant improvement in wound healing being seen at 21 days post-operatively. Histological examinations of healed wounds from all samples showed a thin epidermis plus recovered skin appendages in the dermal layer for samples with cell. Thus, the graft of nanofibrous PCL scaffolds loaded with USSC showed better results during the healing process of skin defects in rat models.

Cellular Response of Limbal Stem Cells on Polycaprolactone Nanofibrous Scaffolds for Ocular Epithelial Regeneration

Purpose: The aim of this study was to develop nanofibrous polycaprolactone (PCL) substrate for limbal stem cell (LSC) expansion that can serve as a potential alternative substrate to replace human amniotic membrane (AM).Materials and methods: The human limbus stem cell was used to evaluate the biocompatibility of substrates (nanofibrous scaffold and, human AM) based on their phenotypic profile, viability, proliferation and attachment ability.Results: Biocompatibility results indicated that the all substrates were highly biocompatible, as LSCs could favorably attach and proliferate on the nanofibrous surface. Microscopic figures showed that the human LSCs were firmly anchored to the substrates and were able to retain a normal corneal stem cell phenotype. Microscopic analyses illustrated that cells infiltrated the nanofibers and successfully formed a three-dimensional corneal epithelium, which was viable for two weeks. Immunocytochemistry (ICC) and real time-PCR results revealed no change in the expression profile of LECs grown on nanofibrous substrate when compared to those grown on human AM.Conclusion: In addition, electrospun nanofibrous PCL substrate provides not only a milieu supporting LSCs expansion, but also serve as a useful alternative carrier for ocular surface tissue engineering and could be used as an alternative substrate to AM.

Vascular Tissue Engineering Using Polycaprolactone Nanofibrous Scaffolds Fabricated via Electrospinning

Electrospinning technique was used for the fabrication of Polycaprolactone (PCL) nanofibrous scaffolds as 2D biodegradable polymer for soft and hard tissue replacement. Nanofiber’s morphology, porosity, phase identification as well as thermal and biological properties were assessed using standard techniques. Bone marrow mesenchymal stem cells (BMSCs-hTERT) were used for investigating properties of PCL scaffolds in the field of tissue engineering. The ability of PCL nanofibrous scaffolds to attach, grow and differentiate the hBMSCs into matrix mineralizing osteoblastic cells was investigated. The SEM micrographs showed beads free and non-agglomerated highly aligned nanofibrous morphology with uniform diameter varied from 200 to 1000 nm depending on the solution concentration and voltage. The EDS results showed that the carbon is the principal element of PCL nanofibers indicating that there are no chemical impurities into PCL polymer. The results of DSC confirmed the semi-crystalline nature of PLC. The TGA results showed that the PCL nanofibers have single stage thermal degradation with no weight loss up to 220 � C, with a slight weight loss from 240‐440 � C due to the combustion of organic PCL matrix. The cell culture results showed that the cells do attach to the scaffold and they are alive and proliferate in the same rate as when cultured on plastic. Moreover the human MSCs differentiated to osteoblasts with calcium deposition when cultured on PCL nanofibrous scaffold, this was confirmed by ALP-, Alezarin staining and by scanning electron microscopy.