Nejvíce citovaný článek - PubMed ID 34067330
Healing and Angiogenic Properties of Collagen/Chitosan Scaffolds Enriched with Hyperstable FGF2-STAB® Protein: In Vitro, Ex Ovo and In Vivo Comprehensive Evaluation
Treatment of complete loss of skin thickness requires expensive cellular materials and limited skin grafts used as temporary coverage. This paper presents an acellular bilayer scaffold modified with polydopamine (PDA), which is designed to mimic a missing dermis and a basement membrane (BM). The alternate dermis is made from freeze-dried collagen and chitosan (Coll/Chit) or collagen and a calcium salt of oxidized cellulose (Coll/CaOC). Alternate BM is made from electrospun gelatin (Gel), polycaprolactone (PCL), and CaOC. Morphological and mechanical analyzes have shown that PDA significantly improved the elasticity and strength of collagen microfibrils, which favorably affected swelling capacity and porosity. PDA significantly supported and maintained metabolic activity, proliferation, and viability of the murine fibroblast cell lines. The in vivo experiment carried out in a domestic Large white pig model resulted in the expression of pro-inflammatory cytokines in the first 1-2 weeks, giving the idea that PDA and/or CaOC trigger the early stages of inflammation. Otherwise, in later stages, PDA caused a reduction in inflammation with the expression of the anti-inflammatory molecule IL10 and the transforming growth factor β (TGFβ1), which could support the formation of fibroblasts. Similarities in treatment with native porcine skin suggested that the bilayer can be used as an implant for full-thickness skin wounds and thus eliminate the use of skin grafts.
- Klíčová slova
- Bilayer, Chitosan, Collagen, Oxidized cellulose, Polydopamine, Wound healing,
- MeSH
- myši MeSH
- nanovlákna * MeSH
- prasata MeSH
- sloučeniny osmia MeSH
- zánět MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chloropentaammineosmium(III) chloride MeSH Prohlížeč
- polydopamine MeSH Prohlížeč
- sloučeniny osmia MeSH
Biomimicking native tissues and organs require the development of advanced hydrogels. The patterning of hydrogel surfaces may enhance the cellular functionality and therapeutic efficacy of implants. For example, nanopatterning of the intraocular lens (IOL) surface can suppress the upregulation of cytoskeleton proteins (actin and actinin) within the cells in contact with the IOL surface and, hence, prevent secondary cataracts causing blurry or opaque vision. Here we introduce a fast and efficient method for fabricating arrays consisting of millions of individual nanostructures on the hydrogel surface. In particular, we have prepared the randomly distributed nanopillars on poly(2-hydroxyethyl methacrylate) hydrogel using replica molding and show that the number, shape, and arrangement of nanostructures are fully adjustable. Characterization by atomic force microscopy revealed that all nanopillars were of similar shape, narrow size distribution, and without significant defects. In imprint lithography, choosing the appropriate hydrogel composition is critical. As hydrogels with imprinted nanostructures mimic the natural cell environment, they can find applications in fundamental cell biology research, e.g., they can tune cell attachment and inhibit or promote cell clustering by a specific arrangement of protrusive nanostructures on the hydrogel surface.
- MeSH
- hydrogely chemie MeSH
- mikroskopie atomárních sil MeSH
- nanostruktury * chemie MeSH
- PEG-DMA hydrogel chemie MeSH
- polyhydroxyethylmethakrylát * chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- hydrogely MeSH
- PEG-DMA hydrogel MeSH
- polyhydroxyethylmethakrylát * MeSH
In this work, the biological properties of three-dimensional scaffolds based on a blend of nanohydroxyapatite (nHA), silk fibroin (SF), and chitosan (CTS), were prepared using a lyophilization technique with various weight ratios: 10:45:45, 15:15:70, 15:70:15, 20:40:40, 40:30:30, and 70:15:15 nHA:SF:CTS, respectively. The basic 3D scaffolds were obtained from 5% (w/w) chitosan and 5% silk fibroin solutions and then nHA was added. The morphology and physicochemical properties of scaffolds were studied and compared. A biological test was performed to study the growth and osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs). It was found that the addition of chitosan increases the resistance properties and extends the degradation time of materials. In vitro studies with human mesenchymal stem cells found a high degree of biotolerance for the materials produced, especially for the 20:40:40 and 15:70:15 (nHa:SF:CTS) ratios. The presence of silk fibroin and the elongated shape of the pores positively influenced the differentiation of cells into osteogenic cells. By taking advantage of the differentiation/proliferation cues offered by individual components, the composites based on the nanohydroxyapatite, silk fibroin, and chitosan scaffold may be suitable for bone tissue engineering, and possibly offer an alternative to the widespread use of collagen materials.
- Klíčová slova
- biological study, bone regeneration, chitosan, hydroxyapatite, mesenchymal stem cells, osteogenesis, silk fibroin,
- Publikační typ
- časopisecké články MeSH
The utilization of poly(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) with entrapped fish oil (FO) loaded in collagen-based scaffolds for cutaneous wound healing using a porcine model is unique for the present study. Full-depth cutaneous excisions (5 × 5 cm) on the pig dorsa were treated with pure collagen scaffold (control, C), empty PLGA NPs (NP), FO, mupirocin (MUP), PLGA NPs with entrapped FO (NP/FO) and PLGA NPs with entrapped MUP (NP/MUP). The following markers were evaluated on days 0, 3, 7, 14 and 21 post-excision: collagen, hydroxyproline (HP), angiogenesis and expressions of the COX2, EGF, COL1A1, COL1A3, TGFB1, VEGFA, CCL5 and CCR5 genes. The hypothesis that NP/FO treatment is superior to FO alone and that it is comparable to NP/MUP was tested. NP/FO treatment increased HP in comparison with both FO alone and NP/MUP (day 14) but decreased (p < 0.05) angiogenesis in comparison with FO alone (day 3). NP/FO increased (p < 0.05) the expression of the CCR5 gene (day 3) and tended (p > 0.05) to increase the expressions of the EGF (day 7, day 14), TGFB1 (day 21) and CCL5 (day 7, day 21) genes as compared with NP/MUP. NP/FO can be suggested as a suitable alternative to NP/MUP in cutaneous wound treatment.
- Klíčová slova
- collagen, cutaneous wounds, cyclooxygenase-2, hydroxyproline, mupirocin, nanoparticles, poly(lactic-co-glycolic) acid, polyunsaturated fatty acids n-3, transforming growth factor,
- MeSH
- epidermální růstový faktor genetika farmakologie MeSH
- hojení ran MeSH
- kolagen metabolismus MeSH
- kopolymer kyseliny glykolové a mléčné farmakologie MeSH
- mupirocin * farmakologie MeSH
- nanočástice * MeSH
- prasata MeSH
- rybí oleje farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- epidermální růstový faktor MeSH
- kolagen MeSH
- kopolymer kyseliny glykolové a mléčné MeSH
- mupirocin * MeSH
- rybí oleje MeSH
Many growth factors have been studied as additives accelerating lumbar fusion rates in different animal models. However, their low hydrolytic and thermal stability both in vitro and in vivo limits their workability and use. In the proposed work, a stabilized vasculogenic and prohealing fibroblast growth factor-2 (FGF2-STAB®) exhibiting a functional half-life in vitro at 37 °C more than 20 days was applied for lumbar fusion in combination with a bioresorbable scaffold on porcine models. An experimental animal study was designed to investigate the intervertebral fusion efficiency and safety of a bioresorbable ceramic/biopolymer hybrid implant enriched with FGF2-STAB® in comparison with a tricortical bone autograft used as a gold standard. Twenty-four experimental pigs underwent L2/3 discectomy with implantation of either the tricortical iliac crest bone autograft or the bioresorbable hybrid implant (BHI) followed by lateral intervertebral fixation. The quality of spinal fusion was assessed by micro-computed tomography (micro-CT), biomechanical testing, and histological examination at both 8 and 16 weeks after the surgery. While 8 weeks after implantation, micro-CT analysis demonstrated similar fusion quality in both groups, in contrast, spines with BHI involving inorganic hydroxyapatite and tricalcium phosphate along with organic collagen, oxidized cellulose, and FGF2- STAB® showed a significant increase in a fusion quality in comparison to the autograft group 16 weeks post-surgery (p = 0.023). Biomechanical testing revealed significantly higher stiffness of spines treated with the bioresorbable hybrid implant group compared to the autograft group (p < 0.05). Whilst histomorphological evaluation showed significant progression of new bone formation in the BHI group besides non-union and fibrocartilage tissue formed in the autograft group. Significant osteoinductive effects of BHI based on bioceramics, collagen, oxidized cellulose, and FGF2-STAB® could improve outcomes in spinal fusion surgery and bone tissue regeneration.
- Klíčová slova
- FGF2, animal model, autograft, biomechanics, ceramic, collagen, histology, lumbar spinal fusion, micro-CT, tissue engineering,
- Publikační typ
- časopisecké články MeSH