Aim: This study evaluates the effect of electrospun dressings in critical sized full-thickness skin defects in rabbits. Materials & methods: Electrospun poly-ε-caprolactone (PCL) and polyvinyl alcohol (PVA) nanofibers were tested in vitro and in vivo. Results: The PCL scaffold supported the proliferation of mesenchymal stem cells, fibroblasts and keratinocytes. The PVA scaffold showed significant swelling, high elongation capacity, limited protein adsorption and stimulation of cells. Nanofibrous dressings improved wound healing compared with the control group in vivo. A change of the PCL dressing every 7 days resulted in a decreased epithelial thickness and type I collagen level in the adhesive group, indicating peeling off of the newly formed tissue. In the PVA dressings, the exchange did not affect healing. Conclusion: The results demonstrate the importance of proper dressing exchange.
- MeSH
- buňky 3T3 MeSH
- hojení ran účinky léků MeSH
- králíci MeSH
- kůže * zranění metabolismus patologie MeSH
- myši MeSH
- nanovlákna chemie MeSH
- obvazy * MeSH
- polyestery * chemie farmakologie MeSH
- polyvinylalkohol chemie farmakologie MeSH
- prasata MeSH
- tkáňová adheziva * chemie farmakologie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Crosslinked 3D porous collagen-polysaccharide scaffolds, prepared by freeze-drying, were modified with bovine platelet lysate (BPL) and evaluated in terms of chemical, physical and biological properties. Natural antibacterial polysaccharides like chitosan, chitin/chitosan-glucan complex and calcium salt of oxidized cellulose (CaOC) incorporated in collagen scaffolds affected not only chemo-physical properties of the composite scaffolds but also improved their biological properties, especially when BPL was presented. Lipophilic BPL formed microspheres in porous scaffolds while reduced by half their swelling ratio. The resistance of collagen sponges to hydrolytic degradation in water depended strongly on chemical crosslinking varying from 60 min to more than one year. According to in-vitro tests, chemically crosslinked scaffolds exhibited a good cellular response, cell-matrix interactions, and biocompatibility of the material. The combination of collagen with natural polysaccharides confirmed a significant positive synergistic effect on cultivation of cells as determined by MTS assay and PicoGreen method, as well as on angiogenesis evaluated by ex ovo Chick Chorioallantoic Membrane (CAM) assay. Contrary, modification only by BLP of pure collagen scaffolds exhibited decreased biocompatibility in comparison to unmodified pure collagen scaffold. We propose that the newly developed crosslinked collagen sponges involving bioactive additives could be used as scaffold for growing cells in systems with low mechanical loading in tissue engineering, especially in dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration.
- MeSH
- buněčná adheze účinky léků MeSH
- buňky 3T3 MeSH
- fibroblasty cytologie účinky léků MeSH
- fyziologická neovaskularizace účinky léků MeSH
- hydrolýza MeSH
- kolagen farmakologie MeSH
- kur domácí MeSH
- myši MeSH
- polysacharidy farmakologie MeSH
- proliferace buněk účinky léků MeSH
- reagencia zkříženě vázaná chemie MeSH
- skot MeSH
- teplota MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury chemie MeSH
- trombocyty metabolismus MeSH
- voda chemie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Cílem příspěvku je ukázat přínosy i omezení histologického hodnocení ovlivnění hojení při implantaci podpůrných biomateriálů. Podáváme přehled a fotodokumentaci histologických přehledných a imunohistochemických metod využitelných pro posouzení žádoucích i nežádoucích účinků biomateriálů při hojení tvrdých i měkkých tkání. Diskutujeme jednak mikroskopické hodnocení hojení s využitím semikvantitativních skórovacích systémů, jednak kvantifikaci složení tkáně pomocí jednotlivých spojitých proměnných, které popisují např. počet buněk v jednotkovém objemu, délky cév či vláknitých struktur matrix, plochy povrchů, plošné a objemové podíly složek tkáně, shlukování a kolokalizaci mikroskopických objektů. Strategie systematického nestranného náhodného výběru vede v histologii k významné redukci variability kvantitativních dat.
Our aim was to show the benefits and limitations of histological assessment of healing supported by implantable biomaterials. We reviewed and showed photographs of the histological and immunohistochemical methods applicable for the assessment of desirable and undesirable effects of biomaterials on the healing of hard and soft tissues. Currently used methods for evaluating the microscopic effects of bioengineered materials on the recipient tissue are reviewed. For histopathological analysis, semiquantitative scoring systems can be used. Alternatively, the main tissue constituents may be quantified using continuous variables giving the numerical densities of cells, lengths of microvessels or connective tissue fibres, area surfaces, area and volumes fractions, or clustering and colocalization of microscopic objects. Using systematic uniform random sampling strategies at the level of tissue blocks, sections, and image fields leads to a reasonable low variability of the quantitative results.
- MeSH
- barvicí látky MeSH
- biokompatibilní materiály * MeSH
- histologie MeSH
- imunohistochemie MeSH
- kosti a kostní tkáň anatomie a histologie MeSH
- kůže anatomie a histologie MeSH
- lidé MeSH
- mikrofotografie MeSH
- molekulární biologie MeSH
- počítačové zpracování obrazu MeSH
- tkáně anatomie a histologie MeSH
- výzkum MeSH
- zobrazování trojrozměrné MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
A three-dimensional scaffold of type I collagen and hydroxyapatite enriched with polycaprolactone nanofibers (Coll/HA/PCL), autologous mesenchymal stem cells (MSCs) in osteogenic media, and thrombocyte-rich solution (TRS) was an optimal implant for bone regeneration in vivo in white rabbits. Nanofibers optimized the viscoelastic properties of the Coll/HA scaffold for bone regeneration. MSCs and TRS in the composite scaffold improved bone regeneration. Three types of Coll/HA/PCL scaffold were prepared: an MSC-enriched scaffold, a TRS-enriched scaffold, and a scaffold enriched with both MSCs and TRS. These scaffolds were implanted into femoral condyle defects 6 mm in diameter and 10-mm deep. Untreated defects were used as a control. Macroscopic and histological analyses of the regenerated tissue from all groups were performed 12 weeks after implantation. The highest volume and most uniform distribution of newly formed bone occurred in defects treated with scaffolds enriched with both MSCs and TRS compared with that in defects treated with scaffolds enriched by either component alone. The modulus of elasticity in compressive testing was significantly higher in the Coll/HA/PCL scaffold than those without nanofibers. The composite Coll scaffold functionalized with PCL nanofibers and enriched with MSCs and TRS appears to be a novel treatment for bone defects.
- MeSH
- hydroxyapatit chemie MeSH
- kolagen chemie MeSH
- králíci MeSH
- kultivované buňky MeSH
- mezenchymální kmenové buňky cytologie metabolismus MeSH
- nanovlákna chemie MeSH
- polyestery chemie MeSH
- regenerace kostí * MeSH
- tkáňové podpůrné struktury chemie MeSH
- trombocyty chemie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Incisional hernia is the most common postoperative complication, affecting up to 20% of patients after abdominal surgery. Insertion of a synthetic surgical mesh has become the standard of care in ventral hernia repair. However, the implementation of a mesh does not reduce the risk of recurrence and the onset of hernia recurrence is only delayed by 2-3 years. Nowadays, more than 100 surgical meshes are available on the market, with polypropylene the most widely used for ventral hernia repair. Nonetheless, the ideal mesh does not exist yet; it still needs to be developed. Polycaprolactone nanofibers appear to be a suitable material for different kinds of cells, including fibroblasts, chondrocytes, and mesenchymal stem cells. The aim of the study reported here was to develop a functionalized scaffold for ventral hernia regeneration. We prepared a novel composite scaffold based on a polypropylene surgical mesh functionalized with poly-ε-caprolactone (PCL) nanofibers and adhered thrombocytes as a natural source of growth factors. In extensive in vitro tests, we proved the biocompatibility of PCL nanofibers with adhered thrombocytes deposited on a polypropylene mesh. Compared with polypropylene mesh alone, this composite scaffold provided better adhesion, growth, metabolic activity, proliferation, and viability of mouse fibroblasts in all tests and was even better than a polypropylene mesh functionalized with PCL nanofibers. The gradual release of growth factors from biocompatible nanofiber-modified scaffolds seems to be a promising approach in tissue engineering and regenerative medicine.
- MeSH
- biokompatibilní materiály * chemie toxicita MeSH
- chirurgické síťky * MeSH
- incizní kýla chirurgie MeSH
- myši MeSH
- nanovlákna * chemie toxicita MeSH
- polyestery * chemie toxicita MeSH
- polypropyleny * chemie toxicita MeSH
- proliferace buněk účinky léků MeSH
- trombocyty cytologie MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem 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.
Incisional hernia affects up to 20% of patients after abdominal surgery. Unlike other types of hernia, its prognosis is poor, and patients suffer from recurrence within 10 years of the operation. Currently used hernia-repair meshes do not guarantee success, but only extend the recurrence-free period by about 5 years. Most of them are nonresorbable, and these implants can lead to many complications that are in some cases life-threatening. Electrospun nanofibers of various polymers have been used as tissue scaffolds and have been explored extensively in the last decade, due to their low cost and good biocompatibility. Their architecture mimics the natural extracellular matrix. We tested a biodegradable polyester poly-ε-caprolactone in the form of nanofibers as a scaffold for fascia healing in an abdominal closure-reinforcement model for prevention of incisional hernia formation. Both in vitro tests and an experiment on a rabbit model showed promising results.
- MeSH
- biomechanika MeSH
- břicho chirurgie MeSH
- buňky 3T3 MeSH
- chirurgické síťky MeSH
- hernie prevence a kontrola MeSH
- histocytochemie MeSH
- hojení ran účinky léků MeSH
- králíci MeSH
- mezibuněčné signální peptidy a proteiny chemie farmakologie terapeutické užití MeSH
- myši MeSH
- nanovlákna chemie terapeutické užití MeSH
- polyestery chemie terapeutické užití MeSH
- polypropyleny chemie terapeutické užití MeSH
- pooperační komplikace prevence a kontrola MeSH
- řízená tkáňová regenerace MeSH
- techniky uzavření břišních poranění přístrojové vybavení MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A wide range of drug-delivery systems are currently attracting the attention of researchers. Nanofibers are very interesting carriers for drug delivery. This is because nanofibers are versatile, flexible, nanobiomimetic and similar to extracellular matrix components, possible to be functionalized both on their surface as well as in their core, and also because they can be produced easily and cost effectively. There have been increasing attempts to use nanofibers in the construction of a range of tissues, including cartilage and bone. Nanofibers have also been favorably engaged as a drug-delivery system in cell-free scaffolds. This short overview is devoted to current applications and to further perspectives of nanofibers as drug-delivery devices in the field of cartilage and bone regeneration, and also in osteochondral reconstruction.
- MeSH
- chrupavka cytologie MeSH
- lidé MeSH
- nanovlákna chemie MeSH
- regenerace kostí fyziologie MeSH
- systémy cílené aplikace léků metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
The aim of the study was to evaluate the effect of a cell-free hyaluronate/type I collagen/fibrin composite scaffold containing polyvinyl alcohol (PVA) nanofibers enriched with liposomes, basic fibroblast growth factor (bFGF) and insulin on the regeneration of osteochondral defects. A novel drug delivery system was developed on the basis of the intake effect of liposomes encapsulated in PVA nanofibers. Time-controlled release of insulin and bFGF improved MSC viability in vitro. Nanofibers functionalized with liposomes also improved the mechanical characteristics of the composite gel scaffold. In addition, time-controlled release of insulin and bFGF stimulated MSC recruitment from bone marrow in vivo. Cell-free composite scaffolds containing PVA nanofibers enriched with liposomes, bFGF, and insulin were implanted into seven osteochondral defects of miniature pigs. Control defects were left untreated. After 12 weeks, the composite scaffold had enhanced osteochondral regeneration towards hyaline cartilage and/or fibrocartilage compared with untreated defects that were filled predominantly with fibrous tissue. The cell-free composite scaffold containing PVA nanofibers, liposomes and growth factors enhanced migration of the cells into the defect, and their differentiation into chondrocytes; the scaffold was able to enhance the regeneration of osteochondral defects in minipigs.
- MeSH
- buněčná diferenciace MeSH
- buňky kostní dřeně cytologie MeSH
- chondrocyty cytologie MeSH
- fibrin chemie MeSH
- fibroblastový růstový faktor 2 aplikace a dávkování MeSH
- inzulin aplikace a dávkování MeSH
- kolagen typu I chemie MeSH
- kyselina hyaluronová chemie MeSH
- liposomy MeSH
- mezenchymální kmenové buňky cytologie MeSH
- miniaturní prasata MeSH
- modul pružnosti MeSH
- nanovlákna aplikace a dávkování chemie MeSH
- polyvinylalkohol chemie MeSH
- prasata MeSH
- regenerace kostí * MeSH
- tkáňové podpůrné struktury MeSH
- viabilita buněk MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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
