Bone and cartilage are tissues of a three-dimensional (3D) nature. Therefore, scaffolds for their regeneration should support cell infiltration and growth in all 3 dimensions. To fulfill such a requirement, the materials should possess large, open pores. Centrifugal spinning is a simple method for producing 3D fibrous scaffolds with large and interconnected pores. However, the process of bone regeneration is rather complex and requires additional stimulation by active molecules. In the current study, we introduced a simple composite scaffold based on platelet adhesion to poly-ε-caprolactone 3D fibers. Platelets were used as a natural source of growth factors and cytokines active in the tissue repair process. By immobilization in the fibrous scaffolds, their bioavailability was prolonged. The biological evaluation of the proposed system in the MG-63 model showed improved metabolic activity, proliferation and alkaline phosphatase activity in comparison to nonfunctionalized fibrous scaffold. In addition, the response of cells was dose dependent with improved biocompatibility with increasing platelet concentration. The results demonstrated the suitability of the system for bone tissue.
- MeSH
- adhezivita trombocytů účinky léků MeSH
- alkalická fosfatasa metabolismus MeSH
- kinetika MeSH
- lékové transportní systémy metody MeSH
- lidé MeSH
- mezibuněčné signální peptidy a proteiny aplikace a dávkování farmakologie MeSH
- nádorové buněčné linie MeSH
- osteoblasty cytologie účinky léků ultrastruktura MeSH
- osteogeneze účinky léků MeSH
- polyestery chemie farmakologie MeSH
- proliferace buněk účinky léků MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury chemie MeSH
- trombocyty účinky léků metabolismus ultrastruktura MeSH
- tvar buňky účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Cell infiltration is a critical parameter for the successful development of 3D matrices for tissue engineering. Application of electrospun nanofibers in tissue engineering has recently attracted much attention. Notwithstanding several of their advantages, small pore size and small thickness of the electrospun layer limit their application for development of 3D scaffolds. Several methods for the pore size and/or electrospun layer thickness increase have been recently developed. Nevertheless, tissue engineering still needs emerging of either novel nanofiber-enriched composites or new techniques for 3D nanofiber fabrication. Forcespinning(®) seems to be a promising alternative. The potential of the Forcespinning(®) method is illustrated in preliminary experiment with mesenchymal stem cells.
OBJECTIVES: We prepared 3D poly (ε-caprolactone) (PCL) nanofibre scaffolds and tested their use for seeding, proliferation, differentiation and migration of mesenchymal stem cell (MSCs). MATERIALS AND METHODS: 3D nanofibres were prepared using a special collector for common electrospinning; simultaneously, a 2D PCL nanofibre layer was prepared using a classic plain collector. Both scaffolds were seeded with MSCs and biologically tested. MSC adhesion, migration, proliferation and osteogenic differentiation were investigated. RESULTS: The 3D PCL scaffold was characterized by having better biomechanical properties, namely greater elasticity and resistance against stress and strain, thus this scaffold will be able to find broad applications in tissue engineering. Clearly, while nanofibre layers of the 2D scaffold prevented MSCs from migrating through the conformation, cells infiltrated freely through the 3D scaffold. MSC adhesion to the 3D nanofibre PCL layer was also statistically more common than to the 2D scaffold (P < 0.05), and proliferation and viability of MSCs 2 or 3 weeks post-seeding, were also greater on the 3D scaffold. In addition, the 3D PCL scaffold was also characterized by displaying enhanced MSC osteogenic differentiation. CONCLUSIONS: We draw the conclusion that all positive effects observed using the 3D PCL nanofibre scaffold are related to the larger fibre surface area available to the cells. Thus, the proposed 3D structure of the nanofibre layer will find a wide array of applications in tissue engineering and regenerative medicine.
- MeSH
- buněčná diferenciace * MeSH
- buněčné kultury přístrojové vybavení metody MeSH
- kultivované buňky MeSH
- lidé MeSH
- mezenchymální kmenové buňky cytologie metabolismus MeSH
- nanovlákna chemie ultrastruktura MeSH
- osteogeneze MeSH
- osteokalcin metabolismus MeSH
- pohyb buněk MeSH
- polyestery chemie MeSH
- povrchové vlastnosti MeSH
- proliferace buněk MeSH
- pružnost MeSH
- regenerativní lékařství MeSH
- sialoprotein vázající integrin metabolismus MeSH
- tkáňové inženýrství MeSH
- tkáňové podpůrné struktury * MeSH
- viabilita buněk MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
PURPOSE OF THE STUDY Articular cartilage defects arise due to injury or osteochondral disease such as osteonecrosis or osteochondritis dissecans. In adult patients cartilage has minimal ability to repair itself and the lesions develop into degenerative arthritis. Overcoming the low regenerative capacity of the cartilage cells and the Hayflick limit poses a challenge for the therapy of osteochondral defects. Composite scaffolds with appropriate biomechanical properties combined with a suitable blend of proliferation and differentiation factors could be a solution. The aim of this in vitro study was to develop a novel functionalised hydrogel with an integrated drug delivery system stimulating articular cartilage regeneration. MATERIAL AND METHODS Injectable collagen/ hyaluronic acid/fibrin composite hydrogel was mixed with nanofibre-based microparticles. These were loaded with ascorbic acid and dexamethasone. In addition, the effect of thrombocyte-rich solution (TRS) was studied. The gels seeded with mesenchymal stem cells (MSCs) were cultivated for 14 days. The viability, proliferation and morphology of the cells were evaluated using molecular and microscopic methods. Scaffold degradation was also assessed. RESULTS The cultivation study showed that MSCs remained viable in all experimental groups, which indicated good biocompatibility of the gel. However, the number of cells in the groups enriched with microparticles was lower than in the other groups. On the other hand, confocal microscopy showed higher cell viability and rounded morphology of the cells, which can be associated with chodrogenic differentiation. The scaffolds containing microparticles showed significantly higher stability during the 14-day experiment. DISCUSSION Our results suggest that the addition of microparticles to the scaffold improved cell differentiation into the chondrogenic lineage, resulting in a lower proliferation rate. Cell viability was better in the groups enriched with microparticles that served as an efficient drug delivery system. In addition, the presence of microparticles slowed down gel degradation which can help achieve sufficient stability of the system for the time frame required for cartilage regeneration. CONCLUSIONS The novel approach described here produced an efficient system where microparticles served as a drug delivery system and stabilised the gel for prolonged periods of time. These characteristics play an important role in the development of scaffolds for cartilage regeneration. In the future the results of these in vitro experiments will be verified in an in vivo study.
- MeSH
- injekce intraartikulární MeSH
- kloubní chrupavka * patologie účinky léků MeSH
- lékové transportní systémy * metody MeSH
- multipotentní kmenové buňky * fyziologie metabolismus MeSH
- nanovlákna terapeutické užití MeSH
- nosiče léků * MeSH
- osteoartróza terapie MeSH
- PEG-DMA hydrogel terapeutické užití MeSH
- příprava léků metody MeSH
- proliferace buněk MeSH
- řízená tkáňová regenerace * metody MeSH
- techniky in vitro MeSH
- trombocyty fyziologie metabolismus MeSH
- Publikační typ
- práce podpořená grantem MeSH
The structural properties of microfiber meshes made from poly(2-hydroxyethyl methacrylate) (PHEMA) were found to significantly depend on the chemical composition and subsequent cross-linking and nebulization processes. PHEMA microfibres showed promise as scaffolds for chondrocyte seeding and proliferation. Moreover, the peak liposome adhesion to PHEMA microfiber scaffolds observed in our study resulted in the development of a simple drug anchoring system. Attached foetal bovine serum-loaded liposomes significantly improved both chondrocyte adhesion and proliferation. In conclusion, fibrous scaffolds from PHEMA are promising materials for tissue engineering and, in combination with liposomes, can serve as a simple drug delivery tool.
- MeSH
- biokompatibilní materiály chemie MeSH
- buněčná adheze MeSH
- chondrocyty cytologie MeSH
- fluorescenční mikroskopie metody MeSH
- konfokální mikroskopie metody MeSH
- lékové transportní systémy MeSH
- liposomy chemie MeSH
- nosiče léků chemie MeSH
- polyhydroxyethylmethakrylát chemie MeSH
- polymery chemie MeSH
- proliferace buněk MeSH
- racionální návrh léčiv MeSH
- reagencia zkříženě vázaná chemie MeSH
- skot MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury chemie MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Hydrogels prepared from a mixture of fibrin and high-molecular weight (MW) hyaluronic acid (HA) were found to be suitable scaffolds for chondrocyte seeding and pig knee cartilage regeneration. Collagen in the hydrogels is not necessary for the formation of biomechanically stable tissue. Regenerated cartilage showed very good biomechanical and histological properties only 6 months after implantation. Notably, the quality of the healing process was dependent on the initial chondrocyte concentration of the scaffolds. These experiments were performed according to good laboratory practice (GLP).
- MeSH
- biokompatibilní materiály chemie MeSH
- biomechanika MeSH
- chondrocyty cytologie fyziologie MeSH
- chondrogeneze MeSH
- chrupavka fyziologie chirurgie MeSH
- fibrin chemie MeSH
- hydrogely MeSH
- kyselina hyaluronová chemie MeSH
- miniaturní prasata MeSH
- prasata MeSH
- protézy a implantáty MeSH
- regenerace MeSH
- testování materiálů MeSH
- tkáňové inženýrství MeSH
- tkáňové podpůrné struktury chemie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Non-woven textile mesh from polyglycolic acid (PGA) was found as a proper material for chondrocyte adhesion but worse for their proliferation. Neither hyaluronic acid nor chitosan nor polyvinyl alcohol (PVA) increased chondrocyte adhesion. However, chondrocyte proliferation suffered from acidic byproducts of PGA degradation. However, the addition of PVA and/or chitosan into a wet-laid non-woven textile mesh from PGA improved chondrocyte proliferation seeded in vitro on the PGA-based composite scaffold namely due to a diminished acidification of their microenvironment. This PVA/PGA composite mesh used in combination with a proper hydrogel minimized the negative effect of PGA degradation without dropping positive parameters of the PGA wet-laid non-woven textile mesh. In fact, presence of PVA and/or chitosan in the PGA-based wet-laid non-woven textile mesh even advanced the PGA-based wet-laid non-woven textile mesh for chondrocyte seeding and artificial cartilage production due to a positive effect of PVA in such a scaffold on chondrocyte proliferation.
- MeSH
- buněčná adheze MeSH
- buněčné dělení MeSH
- chondrocyty cytologie MeSH
- chrupavka cytologie MeSH
- financování organizované MeSH
- konfokální mikroskopie MeSH
- králíci MeSH
- kyselina hyaluronová MeSH
- kyselina polyglykolová MeSH
- PEG-DMA hydrogel MeSH
- polyvinylalkohol MeSH
- techniky tkáňových kultur metody MeSH
- textilie MeSH
- tkáňové podpůrné struktury MeSH
- voda MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
