Most cited article - PubMed ID 23499757
A cell-free nanofiber composite scaffold regenerated osteochondral defects in miniature pigs
The skin is the primary tissue affected by wounds and aging, significantly impacting its protective function. Natural products are widely used in cosmetics, representing a new approach to preventing age-related damage. Nanomedicine combines nanotechnology and traditional treatments to create innovative drugs. The main targets of nanotechnological approaches are wound healing, regeneration, and rejuvenation of skin tissue. The skin barrier is not easily permeable, and the creation of modern nanodevices is a way to improve the passive penetration of substances. In this study, Helichrysum italicum oil (HO) was combined with different types of electrospun nanofibers to study their protective activity on the skin and to evaluate their future application for topical treatments. In the present research, we used biodegradable polymers, including polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP), which were characterized by a scanning electron microscope (SEM). All results show a positive trend in cell proliferation and viability of human skin stem cells (SSCs) and BJ fibroblasts pre-treated with combined nanofibers and then exposed to UV stress. Gene expression analysis revealed the activation of a molecular rejuvenation program in SSCs treated with functionalized nanofibers before UV exposure. Understanding the mechanisms involved in skin changes during aging allows for the future application of nanomaterials combined with HO directly to the patients.
- Keywords
- Helichrysum italicum, bioactive molecules, drug delivery, molecular mechanisms, nanofibers, nanosystem, skin aging, stem cells,
- MeSH
- Biological Products * pharmacology MeSH
- Wound Healing MeSH
- Skin MeSH
- Humans MeSH
- Nanofibers * MeSH
- Polyvinyl Alcohol MeSH
- Skin Aging * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Biological Products * MeSH
- Polyvinyl Alcohol MeSH
Scaffolds made of degradable polymers, such as collagen, polyesters or polysaccharides, are promising matrices for fabrication of bioartificial vascular grafts or patches. In this study, collagen isolated from porcine skin was processed into a gel, reinforced with collagen particles and with incorporated adipose tissue-derived stem cells (ASCs). The cell-material constructs were then incubated in a DMEM medium with 2% of FS (DMEM_part), with added polyvinylalcohol nanofibers (PVA_part sample), and for ASCs differentiation towards smooth muscle cells (SMCs), the medium was supplemented either with human platelet lysate released from PVA nanofibers (PVA_PL_part) or with TGF-β1 + BMP-4 (TGF + BMP_part). The constructs were further endothelialised with human umbilical vein endothelial cells (ECs). The immunofluorescence staining of alpha-actin and calponin, and von Willebrand factor, was performed. The proteins involved in cell differentiation, the extracellular matrix (ECM) proteins, and ECM remodelling proteins were evaluated by mass spectrometry on day 12 of culture. Mechanical properties of the gels with ASCs were measured via an unconfined compression test on day 5. Gels evinced limited planar shrinkage, but it was higher in endothelialised TGF + BMP_part gel. Both PVA_PL_part samples and TGF + BMP_part samples supported ASC growth and differentiation towards SMCs, but only PVA_PL_part supported homogeneous endothelialisation. Young modulus of elasticity increased in all samples compared to day 0, and PVA_PL_part gel evinced a slightly higher ratio of elastic energy. The results suggest that PVA_PL_part collagen construct has the highest potential to remodel into a functional vascular wall.
- Keywords
- adipose tissue-derived stem cells, collagen particles, endothelial cells, extracellular matrix, gel reinforcement, remodelling, stem cells differentiation, tissue engineering, vascular patches,
- MeSH
- Cell Differentiation MeSH
- Human Umbilical Vein Endothelial Cells MeSH
- Extracellular Matrix Proteins metabolism MeSH
- Gels metabolism MeSH
- Stem Cells metabolism MeSH
- Collagen * metabolism MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Myocytes, Smooth Muscle metabolism MeSH
- Swine MeSH
- Tissue Engineering methods MeSH
- Adipose Tissue * MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Extracellular Matrix Proteins MeSH
- Gels MeSH
- Collagen * MeSH
Hydrogels are suitable for osteochondral defect regeneration as they mimic the viscoelastic environment of cartilage. However, their biomechanical properties are not sufficient to withstand high mechanical forces. Therefore, we have prepared electrospun poly-ε-caprolactone-chitosan (PCL-chit) and poly(ethylene oxide)-chitosan (PEO-chit) nanofibers, and FTIR analysis confirmed successful blending of chitosan with other polymers. The biocompatibility of PCL-chit and PEO-chit scaffolds was tested; fibrochondrocytes and chondrocytes seeded on PCL-chit showed superior metabolic activity. The PCL-chit nanofibers were cryogenically grinded into microparticles (mean size of about 500 µm) and further modified by polyethylene glycol-biotin in order to bind the anti-CD44 antibody, a glycoprotein interacting with hyaluronic acid (PCL-chit-PEGb-antiCD44). The PCL-chit or PCL-chit-PEGb-antiCD44 microparticles were mixed with a composite gel (collagen/fibrin/platelet rich plasma) to improve its biomechanical properties. The storage modulus was higher in the composite gel with microparticles compared to fibrin. The Eloss of the composite gel and fibrin was higher than that of the composite gel with microparticles. The composite gel either with or without microparticles was further tested in vivo in a model of osteochondral defects in rabbits. PCL-chit-PEGb-antiCD44 significantly enhanced osteogenic regeneration, mainly by desmogenous ossification, but decreased chondrogenic differentiation in the defects. PCL-chit-PEGb showed a more homogeneous distribution of hyaline cartilage and enhanced hyaline cartilage differentiation.
- Keywords
- CD44 antibody, cartilage, collagen, fibrin, microparticles, poly-ε-caprolactone,
- Publication type
- Journal Article MeSH
Bone regeneration is a long-term process requiring proper scaffolding and drug delivery systems. The current study delivers a three-dimensional (3D) scaffold prepared by blend centrifugal spinning loaded with the osteogenic supplements (OS) β-glycerol phosphate, ascorbate-2-phosphate and dexamethasone. The OS were successfully encapsulated into a fibrous scaffold and showed sustained release for 30 days. Furthermore, biological testing showed the osteoinductive properties of the scaffolds on a model of human mesenchymal stem cells and stimulatory effect on a model of osteoblasts. The osteoinductive properties were further proved in vivo in critical size defects of rabbits. The amount of bone trabecules was bigger compared to control fibers without OS. The results indicate that due to its long-term drug releasing properties, single step fabrication process and 3D structure, the system shows ideal properties for use as a cell-free bone implant in tissue-engineering.
- Publication type
- Journal Article MeSH
OBJECTIVES: 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). MATERIALS AND METHODS: 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: 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. CONCLUSION: 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.
- Keywords
- bioactive peptides, electrospun scaffold, mesenchymal stem cells, osteogenic differentiation,
- MeSH
- Cell Differentiation drug effects MeSH
- Cell Culture Techniques MeSH
- Collagen Type II genetics metabolism MeSH
- Collagen Type I genetics metabolism MeSH
- Microscopy, Confocal MeSH
- Cells, Cultured MeSH
- Mesenchymal Stem Cells cytology drug effects metabolism MeSH
- Microscopy, Electron, Scanning MeSH
- Swine, Miniature MeSH
- Osteogenesis drug effects MeSH
- Osteocalcin genetics metabolism MeSH
- Peptides chemistry pharmacology MeSH
- Swine MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Collagen Type II MeSH
- Collagen Type I MeSH
- Osteocalcin MeSH
- Peptides MeSH
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.
- Keywords
- growth factors, hernia regeneration, in vivo, nanofibers, surgical mesh,
- MeSH
- Biomechanical Phenomena MeSH
- Abdomen surgery MeSH
- 3T3 Cells MeSH
- Surgical Mesh MeSH
- Hernia prevention & control MeSH
- Histocytochemistry MeSH
- Wound Healing drug effects MeSH
- Rabbits MeSH
- Intercellular Signaling Peptides and Proteins chemistry pharmacology therapeutic use MeSH
- Mice MeSH
- Nanofibers chemistry therapeutic use MeSH
- Polyesters chemistry therapeutic use MeSH
- Polypropylenes chemistry therapeutic use MeSH
- Postoperative Complications prevention & control MeSH
- Guided Tissue Regeneration MeSH
- Abdominal Wound Closure Techniques instrumentation MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Intercellular Signaling Peptides and Proteins MeSH
- polycaprolactone MeSH Browser
- Polyesters MeSH
- Polypropylenes MeSH