RESUMEN
The periosteum plays a vital role in the regeneration of critical-size bone defects and highly comminuted fractures, promoting the differentiation of osteoblasts, accelerating the reconstruction of the vascular network, and guiding bone tissue regeneration. However, the materials loaded with exogenous growth factors are limited by the release and activity of the elements. Therefore, the material structure must be carefully designed for the periosteal function. Here, a self-adaptive biomimetic periosteum strategy is proposed, which is a novel interpenetrating double network hydrogel consisting of diselenide-containing gelatin and calcium alginate (modified natural collagen and polysaccharide) to enhance the stability, anti-swelling, and delayed degradation of the hydrogel. The diselenide bond continuously releases nitric oxide (NO) by metabolizing endogenous nitrosated thiols (RSNO), activates the nitric oxide-cycle guanosine monophosphate (NO-cGMP) signal pathway, coordinates the coupling effect of angiogenesis and osteogenesis, and accelerates the repair of bone defects. This self-adaptive biomimetic periosteum with the interpenetrating double network structure formed by the diselenide-containing gelatin and calcium alginate has been proven to be safe and effective in repairing critical-size bone defects and is expected to provide a promising strategy for solving clinical problems.
Asunto(s)
Óxido Nítrico , Periostio , Periostio/química , Óxido Nítrico/análisis , Gelatina/farmacología , Gelatina/química , Biomimética , Angiogénesis , Regeneración Ósea , Osteogénesis , Alginatos , Hidrogeles/química , Andamios del Tejido/química , Ingeniería de TejidosRESUMEN
An artificial periosteum has emerged as an encouraging candidate for bone defect repair. Currently, it remains a great challenge to develop a multifaceted biomimetic periosteum integrating multifunctional features of bioactivities and unique mechanical properties. Here, we successfully fabricated an artificial periosteum (AP) composed of hierarchically assembled Mg-doped mineralized collagen microfibrils with a biomimetically rotated lamellar structure via a "multiscale cascade regulation" strategy combining multiple techniques such as molecular self-assembly, electrospinning, and pressure-driven fusion from molecular to macroscopic levels. The AP has excellent mechanical properties with an ultimate strength and a tensile modulus of 15.9 MPa and 1.1 GPa, respectively. The involvement of Mg-doped nano-hydroxyapatite endowed the AP with good osteogenic and angiogenic activities to promote osteogenic differentiation of bone marrow mesenchymal stem cells and human umbilical vein endothelial cell differentiation into capillary-like structures in vitro. In addition, the results of in vivo evaluations in a rat cranial bone defect model including micro-CT morphology, histological staining, and immunohistochemical analysis showed that Mg-doped mineralized collagen-based AP (MgMC@AP) significantly facilitated cranial bone regeneration and fast vascularization. Our findings suggest that the AP mimicked the composition, lamellar structure, mechanical properties, and biological activities of natural periosteum/lamellae, showing great promise for bone tissue regeneration.
Asunto(s)
Osteogénesis , Periostio , Ratas , Animales , Humanos , Periostio/química , Andamios del Tejido/química , Biomimética , Regeneración Ósea , Colágeno , Cráneo , Ingeniería de Tejidos/métodosRESUMEN
BACKGROUND AND AIM: Membranes for guided bone regeneration should have a mechanical structure and a chemical composition suitable for mimicking biological structures. In this work, we pursue the development of periosteum-inspired bilayered membranes obtained by crosslinking alginate with different amounts of nanohydroxyapatite. EXPERIMENTS: Alginate-nanohydroxyapatite interaction was studied by rheology and infrared spectroscopy measurements. The membranes were characterized regarding their tensile strength, degradation and surface morphology. Finally, cell cultures were performed on each side of the membranes. FINDINGS: The ionic bonding between alginate polysaccharide networks and nanohydroxyapatite was proven, and had a clear effect in the strength and microstructure of the hydrogels. Distinct surface characteristics were achieved on each side of the membranes, resulting in a highly porous fibrous side and a mineral-rich side with higher roughness and lower porosity. Moreover, the effect of amount of nanohydroxyapatite was reflected in a decrease of the membranes' plasticity and an increment of degradation rate. Finally, it was proved that osteoblast-like cells proliferated and differentiated on the mineral-rich side, specially when a higher amount of nanohydroxyapatite was used, whereas fibroblasts-like cells were able to proliferate on the fibrous side. These periosteum-inspired membranes are promising biomaterials for guided tissue regeneration applications.
Asunto(s)
Alginatos/química , Materiales Biomiméticos/química , Durapatita/química , Membrana Dobles de Lípidos/química , Nanopartículas/química , Periostio/química , Diferenciación Celular , Células Cultivadas , Humanos , Osteoblastos/química , Osteoblastos/citología , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
The process of bone formation onto the bone surface using a hydroxyapatite/collagen bone-like nanocomposite (HAp/Col) was investigated. Immersion tests were performed to evaluate the impact of pH on the degradation of the specimens in an aqueous environment. The specimens were soaked in aqueous solutions of pH 4.0, 5.0, and 7.0. Using standardized images, the top-view areas of the specimens were measured. Animal experiments were performed to investigate the bone formation process onto the bone surface. The specimens were placed under the rat calvarial periosteum, and µCT image analysis and histological observation were performed on samples harvested on postoperative Days 3, 5, and 7. In all experiments, ß-tricalciumphosphate (ß-TCP) was adopted as the control. HAp/Col turned to gel in acidic environments below pH 5.0. In contrast to the ß-TCP, the HAp/Col specimens placed under the periosteum expanded and attained a hollow structure with a gel-filled center, accompanied by larger volume of new bone and appearance of TRAP-positive multinucleated cells on postoperative Day 5. Therefore, HAp/Col can enhance bone formation onto the bone surface via induction of TRAP-positive multinucleated cells, and may have clinical applications.
Asunto(s)
Sustitutos de Huesos/química , Colágeno/química , Durapatita/química , Nanocompuestos/química , Periostio/química , Andamios del Tejido/química , Animales , Refuerzo Biomédico , Regeneración Ósea , Colágeno/metabolismo , Durapatita/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Osteogénesis , Periostio/metabolismo , Porosidad , Ratas , Ingeniería de Tejidos , Microtomografía por Rayos XRESUMEN
Despite high biocompatibility of titanium and its alloys, this metal causes various side effects in the human body. It is believed that titanium biomaterials may induce an innate/adaptive immune response. However, still little is known about changes caused by titanium mandible implants, particularly with regard to bone healing. The latest studies showed disturbances in the antioxidant barrier, increased oxidative/nitrosative stress, as well as mitochondrial abnormalities in the periosteum covering titanium mandible fixations; nevertheless, the impact of titanium implants on free radical production, inflammation, and mandible apoptosis are still unknown. Because severe inflammation and apoptosis are among the main factors responsible for disturbances in osteointegration as well as implant rejection, this study is the first to evaluate pro-oxidant enzymes, cytokines as well as pro- and anti-apoptotic proteins in the periosteum of patients with a broken jaw, treated with titanium miniplates and miniscrews. The study group consisted of 29 patients with double-sided fracture of the mandible body requiring surgical treatment. We found significantly higher activity of NADPH oxidase and xanthine oxidase as well as enhanced rate of free radical production in the periosteum of patients in the study group compared to the control group. The markers of inflammation [interleukin 1 (IL-1), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), transforming growth factor ß (TGF-ß) and ß-glucuronidase (GLU)] as well as apoptosis [Bax, Bax/Bcl-2 ratio, caspase-3 (CAS-3) and nitric oxide (NO)] were significantly elevated in periosteum covering titanium fixations compared to the control group. In the study group, we also demonstrated an increased content of titanium on the periosteum surface, which positively correlated with CAS-3 activity. The study led us to the conclusion that titanium mandible implants increase the production of pro-inflammatory cytokines, and enhance free radical generation in the periosteum covering titanium miniplates and miniscrews. Additionally, exposure to Ti6Al4V titanium alloy induces apoptosis in the mandible periosteum. However, no clinical signs of the said phenomena have been observed.
Asunto(s)
Fracturas Óseas/terapia , Traumatismos Mandibulares/terapia , Prótesis Mandibular/efectos adversos , Titanio/efectos adversos , Adulto , Apoptosis , Caspasa 3/metabolismo , Citocinas/metabolismo , Femenino , Fracturas Óseas/metabolismo , Fracturas Óseas/patología , Radicales Libres/metabolismo , Humanos , Inflamación/metabolismo , Masculino , Traumatismos Mandibulares/metabolismo , Traumatismos Mandibulares/patología , NADPH Oxidasas/metabolismo , Periostio/química , Periostio/metabolismo , Periostio/patología , Titanio/análisis , Xantina Oxidasa/metabolismo , Adulto JovenRESUMEN
The traditional Chinese medicine icariin (ICA) and broad-spectrum antibacterial drug moxifloxacin hydrochloride (MOX) were introduced into a polycaprolactone core and gelatin shell, respectively, to develop osteogenic and antibacterial biomimetic periosteum by coaxial electrospinning. The physical properties, drug release, degradation, antibacterial property, in vitro and in vivo osteogenesis performances were investigated. Results demonstrated that stepwise and controlled drug release profiles were achieved based on the core-shell configuration and disparate degradation rate of PCL and gelatin. Only 20% ICA was released from this dual drug-loaded membrane after 1 month while the release of MOX was almost completed. Moreover, clear in vitro antibacterial effect and enhancement in osteogenic marker expressions including osteocalcin, type-I collagen expression, and calcium deposition were observed. Notably, the dual drug-loaded membrane displayed fascinating properties contributing to in vivo bone formation in terms of quality and quantity in a rabbit radius defect model.
Asunto(s)
Antibacterianos/administración & dosificación , Medicamentos Herbarios Chinos/administración & dosificación , Flavonoides/administración & dosificación , Moxifloxacino/administración & dosificación , Nanofibras/química , Poliésteres/química , Animales , Antibacterianos/farmacología , Materiales Biocompatibles/química , Materiales Biomiméticos/química , Regeneración Ósea/efectos de los fármacos , Línea Celular , Medicamentos Herbarios Chinos/farmacología , Flavonoides/farmacología , Gelatina/química , Membranas Artificiales , Ratones , Moxifloxacino/farmacología , Nanofibras/ultraestructura , Osteogénesis/efectos de los fármacos , Periostio/química , Conejos , Andamios del Tejido/químicaRESUMEN
Deer antlers, the secondary organs of deer, are a unique model to study regeneration of organ/tissue in mammals. Pedicle periosteum (PP) is the key tissue type for antler regeneration. Based on our previous study, the DNA methylation was found to be the basic molecular mechanism underlying the antler regeneration. In this study, we compare the genome-wide DNA methylation level in regenerative tissues (the potentiated PP of antler, muscle, heart and liver) and non-regenerative tissue (the dormant PP) of deer by the fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) method. Our results showed that DNA methylation level was significantly lower in the regenerative tissues compared to the non-regenerative tissue (Pâ¯<â¯0.05). Furthermore, 26 T-DMRs which displayed different methylated status in regenerative and non-regenerative tissues were identified by the MSAP method, and were further confirmed by Southern blot analysis. Taken together, our data suggest that DNA methylation, an important epigenetic regulation mechanism, may play an important role in the mammalian tissue/organ regeneration.
Asunto(s)
Metilación de ADN , Ciervos/genética , Regeneración , Análisis de Secuencia de ADN/métodos , Animales , Cuernos de Venado/química , Cuernos de Venado/fisiología , Epigénesis Genética , Hígado/química , Hígado/fisiología , Músculo Esquelético/química , Músculo Esquelético/fisiología , Miocardio/química , Especificidad de Órganos , Periostio/química , Periostio/fisiologíaRESUMEN
Mandibular distraction is a surgical process that progressively lengthens bone. To improve the distraction procedure and devices, the load of distraction and the mechanical strain of soft tissues during the process must be determined. We tested the assumption that it could be the periosteum primarily opposing distraction. Therefore we assessed the mechanical properties of the human mandibular periosteum and compared the stress-strain data with the torque measured on the activator during a cadaveric mandibular distraction. A 20 mm horizontal mandibular distraction was performed in 7 cadavers using standard distractors. Torque was measured with a torquemeter placed on the activation rods of the devices, providing a load (Lt) for each millimeter of distraction. In parallel, 18 periosteum samples were harvested from 9 cadaver mandibles. Uniaxial tensile tests were performed on the specimens and an estimated load (Lc) was calculated using periosteal stress-strain data and mandibular dimensions. During the distraction process, we observed an increase of the load Lt from 11.6 to 50.6 N. The periosteum exhibited a nonlinear viscoelastic stress-strain relationship, typical of biological tissues composed of collagen and elastin. The median Lc and Lt were not significantly different for the first millimeter of distraction. We demonstrated the periosteum is primarily responsible for opposing the distraction load.
Asunto(s)
Mandíbula/química , Mandíbula/cirugía , Periostio/química , Periostio/cirugía , Estrés Mecánico , Femenino , Humanos , MasculinoRESUMEN
Due to the significant role of the periosteum in bone regeneration, we hypothesised that using a specially engineered artificial periosteum could lead to an enhancement in osteogenesis in bone grafts. Herein, we describe our work aimed at fabricating an electrospun fibrous membrane as an artificial periosteum that exhibits flexibility, permeability and osteoinduction. This membrane was designed to cover the complex surface of bone grafts to facilitate and accelerate bone regeneration. The traditional Chinese medicine icariin (ICA) was subsequently introduced into poly (ε-caprolactone) (PCL) /gelatin nanofibers to fabricate an artificial periosteum for the first time. The effects of ICA content on morphology, physical properties, drug release profile, in vitro degradability, biocompatibility and osteogenic differentiation properties were investigated. The ICA-loaded electrospun membranes showed significant improvement in hydrophilicity, high mechanical strength, appropriate degradation rates and excellent biocompatibility. Furthermore, clear enhancement in alkaline phosphatase (ALP) activity, as well as an increase in osteocalcin (OCN) and type collagen I (COL I) expression in MC3T3-E1 cells were detected. Furthermore, we observed clear calcium deposition content in MC3T3-E1 cells cultured on ICA-loaded fibrous matrix. The membrane loaded with 0.05â¯wt.% ICA displayed properties contributing to cell attachment, proliferation and differentiation. These results indicate the huge potential of this ICA-loaded PCL/gelatin electrospun membrane as a biomimetic artificial periosteum to accelerate bone regeneration.
Asunto(s)
Flavonoides/química , Gelatina/química , Nanofibras/química , Periostio/química , Poliésteres/química , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
Cell-mediated mineralization is essential for bone formation and regeneration. In this study, it is proven that extracellular matrix (ECM) of decellularized periosteum can play an initiative and independent role in bone-like apatite formation. Using decellularized periosteum scaffold, it is revealed that ECM scaffold itself can promote critical bone defect regeneration and nude mouse ectopic ossification. The natural collagen matrix of decellularized periosteum can serve as a 3D structural template for Ca-P nuclei initiation, controlling the size and orientation of bone-like mineral crystals. The naturally cross-linked and highly ordered 3D fibrillar network of decellularized periosteum not only provides a model for mimicking mineralization in vitro and in vivo to elucidate the important functions of ECM in bone formation and regeneration, but also can be a promising biomaterial for bone tissue engineering and clinical application.
Asunto(s)
Regeneración Ósea/fisiología , Sustitutos de Huesos/química , Matriz Extracelular/química , Periostio/química , Animales , Enfermedades Óseas/terapia , Regeneración Ósea/efectos de los fármacos , Sustitutos de Huesos/farmacología , Sustitutos de Huesos/uso terapéutico , Calcificación Fisiológica/efectos de los fármacos , Colágeno Tipo I/química , Ratones , Ratones Desnudos , Periostio/citología , Periostio/metabolismo , Ingeniería de Tejidos , Andamios del Tejido/químicaRESUMEN
A novel immunohistochemistry (IHC) approach has been developed to label and localize osterix, a bone-specific transcription factor, within formalin-fixed, paraffin-embedded, tissue-engineered constructs uniquely containing synthetic polymers and human periosteal tissue. Generally, such specimens consisting in part of polymeric materials and mineral are particularly difficult for IHC identification of proteins. Samples here were fabricated from human periosteum, electrospun poly-l-lactic acid (PLLA) nanofibers, and polycaprolactone/poly-l-lactic acid (PCL/PLLA, 75/25) scaffolds and harvested following 10 weeks of implantation in athymic mice. Heat-induced and protease-induced epitope retrieval methods from selected existing protocols were examined to identify osterix. All such protease-induced techniques were unsuccessful. Heat-induced retrieval gave positive results for osterix immunohistochemical staining in sodium citrate/EDTA/Tween 20 with high heat (120C) and pressure (~30 psi) for 10 min, but the heat and pressure levels resulted in tissue damage and section delamination from slides that limited protocol effectiveness. Heat-induced epitope retrieval led to other osterix-positive staining results achieved with minimal impact on structural integrity of the tissue and polymers in sodium citrate/EDTA/Tween 20 buffer at 60C and normal pressure (14.5 psi) for 72 hr. The latter approach identified osterix-positive cells by IHC within periosteal tissue, layers of electrospun PLLA nanofibers, and underlying PCL/PLLA scaffolds of the tissue-engineered constructs.
Asunto(s)
Sustitutos de Huesos/química , Inmunohistoquímica/métodos , Periostio/química , Poliésteres/química , Ingeniería de Tejidos/métodos , Factores de Transcripción/análisis , Animales , Calor , Humanos , Ratones Desnudos , Nanofibras/química , Factor de Transcripción Sp7 , Andamios del Tejido/químicaRESUMEN
Heterotopic ossification (HO) is the abnormal formation of bone in soft tissues and is a frequent complication of hip replacement surgery. Heterotopic ossifications are described to develop via endochondral ossification and standard treatment is administration of indomethacin. It is currently unknown how indomethacin influences heterotopic ossification on a molecular level; therefore, we aimed to determine whether indomethacin might influence heterotopic ossification via impairing the chondrogenic phase of endochondral ossification. Progenitor cell models differentiating in the chondrogenic lineage (ATDC5, primary human bone marrow stem cells and ex vivo periosteal agarose cultures) were treated with increasing concentrations of indomethacin and a decrease in gene- and protein expression of chondrogenic and hypertrophic markers (measured by RT-qPCR and immunoblotting) as well as decreased glycosamino-glycan content (by alcian blue histochemistry) was observed. Even when hypertrophic differentiation was provoked, the addition of indomethacin resulted in decreased hypertrophic marker expression. Interestingly, when mature chondrocytes were treated with indomethacin, a clear increase in collagen type 2 expression was observed. Similarly, when ATDC5 cells and bone marrow stem cells were pre-differentiated to obtain a chondrocyte phenotype and indomethacin was added from this time point onward, low concentrations of indomethacin also resulted in increased chondrogenic differentiation. Indomethacin induces differential effects on in vitro endochondral ossification, depending on the chondrocyte's differentiation stage, with complete inhibition of chondrogenic differentiation as the most pronounced action. This observation may provide a rational behind the elusive mode of action of indomethacin in the treatment of heterotopic ossifications. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:847-857, 2017.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Condrocitos/citología , Indometacina/farmacología , Fosfatasa Alcalina/química , Artroplastia de Reemplazo de Cadera , Células de la Médula Ósea/citología , Diferenciación Celular , Linaje de la Célula , Proliferación Celular , Células Cultivadas , Condrogénesis/efectos de los fármacos , Proteínas de Unión al ADN/metabolismo , Dinoprostona/metabolismo , Glicosaminoglicanos/química , Humanos , Osteogénesis , Periostio/química , Reacción en Cadena en Tiempo Real de la Polimerasa , Células Madre/citología , Células Madre/efectos de los fármacos , Factores de Transcripción/metabolismoRESUMEN
Critical-sized long bone defects suffer from complications including impaired healing and non-union due to substandard healing and integration of devitalized bone allograft. Removal of the periosteum contributes to the limited healing of bone allografts. Restoring a periosteum on bone allografts may provide improved allograft healing and integration. This article reports a polysaccharide-based tissue engineered periosteum that delivers basic fibroblast growth factor (FGF-2), transforming growth factor-ß1 (TGF-ß1), and adipose-derived mesenchymal stem cells (ASCs) to a critical-sized mouse femur defect. The tissue engineered periosteum was evaluated for improving bone allograft healing and incorporation by locally delivering FGF-2, TGF-ß1, and supporting ASCs transplantation. ASCs were successfully delivered and longitudinally tracked at the defect site for at least 7 days post operation with delivered FGF-2 and TGF-ß1 showing a mitogenic effect on the ASCs. At 6 weeks post implantation, data showed a non-significant increase in normalized bone callus volume. However, union ratio analysis showed a significant inhibition in allograft incorporation, confirmed by histological analysis, due to loosening of the nanofiber coating from the allograft surface. Ultimately, this investigation shows our tissue engineered periosteum can deliver FGF-2, TGF-ß1, and ASCs to a mouse critical-sized femur defect and further optimization may yield improved bone allograft healing. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 900-911, 2017.
Asunto(s)
Tejido Adiposo/metabolismo , Sistemas de Liberación de Medicamentos/métodos , Fémur , Factor 2 de Crecimiento de Fibroblastos , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/metabolismo , Periostio/química , Factor de Crecimiento Transformador beta1 , Aloinjertos , Animales , Femenino , Fémur/lesiones , Fémur/metabolismo , Factor 2 de Crecimiento de Fibroblastos/química , Factor 2 de Crecimiento de Fibroblastos/farmacología , Ratones , Ratones Endogámicos BALB C , Ingeniería de Tejidos/métodos , Factor de Crecimiento Transformador beta1/química , Factor de Crecimiento Transformador beta1/farmacologíaRESUMEN
Bone grafting or bone substitute is typically used to bridge a bone defect that has been caused by trauma, tumor resection, pathological degeneration, or congenital deformations. However, bone graft healing and remodeling is always a major concern of orthopedic surgeons. Because the periosteum has a remarkable regenerative capacity and is widely recognized to be essential for the initiation of bone graft healing and remodeling, the present study aimed to produce a rabbit decellularized periosteum (D-periosteum) to be used as a biologic scaffold for future bone tissue engineering. We obtained the D-periosteum by employing a combination of commonly used decellularization processes, which include physical methods as well as chemical and enzymatic solutions. The cellular components were effectively removed, and this removal was demonstrated using current decellularization criteria (H&E staining, DAPI staining, DNA quantification and agarose gel electrophoresis); however, there were no significant alterations of the native extracellular matrix (ECM) properties (collagen, glycosaminoglycan (GAG), microarchitecture and mechanical properties). Periosteum-derived cells (PDCs) could adhere, proliferate and infiltrate into the D-periosteum in vitro. The allogenic D-periosteum was implanted subcutaneously into the backs of rabbits over 28 days to study the biocompatibility in vivo. The D-periosteum did not elicit a severe immunogenic response. In summary, a biologic scaffold composed of ECM from periosteum has been successfully developed. The D-periosteum maintains biocompatibility in vitro and in vivo and, therefore, can provide a naturally compatible scaffold for use in future bone tissue engineering.
Asunto(s)
Sustitutos de Huesos/síntesis química , Trasplante Óseo/instrumentación , Periostio/química , Ingeniería de Tejidos/instrumentación , Andamios del Tejido , Animales , Sistema Libre de Células , Diseño de Equipo , Análisis de Falla de Equipo , Femenino , Ensayo de Materiales , Conejos , Resistencia a la TracciónRESUMEN
BACKGROUND AND OBJECTIVE: The jaw bone, unlike most other bones, is derived from neural crest stem cells, so we hypothesized that it may have different characteristics to bones from other parts of the body, especially in the nature of its periosteum. The periosteum exhibits osteogenic potential and has received considerable attention as a grafting material for the repair of bone and joint defects. MATERIAL AND METHODS: Gene expression profiles of jaw bone and periosteum were evaluated by DNA microarray and real-time polymerase chain reaction. Furthermore, we perforated an area 2 mm in diameter on mouse frontal and parietal bones. Bone regeneration of these calvarial defects was evaluated using microcomputed tomography and histological analysis. RESULTS: The DNA microarray data revealed close homology between the gene expression profiles within the ilium and femur. The gene expression of Wnt-1, SOX10, nestin, and musashi-1 were significantly higher in the jaw bone than in other locations. Microcomputed tomography and histological analysis revealed that the jaw bone had superior bone regenerative abilities than other bones. CONCLUSION: Jaw bone periosteum exhibits a unique gene expression profile that is associated with neural crest cells and has a positive influence on bone regeneration when used as a graft material to repair bone defects. A full investigation of the biological and mechanical properties of jaw bone as an alternative graft material for jaw reconstructive surgery is recommended.
Asunto(s)
Mandíbula/crecimiento & desarrollo , Maxilar/crecimiento & desarrollo , Periostio/crecimiento & desarrollo , Animales , Desarrollo Óseo/genética , Enfermedades Óseas/cirugía , Regeneración Ósea/genética , Trasplante Óseo/métodos , Fémur/química , Hueso Frontal/patología , Hueso Frontal/cirugía , Perfilación de la Expresión Génica , Ilion/química , Masculino , Mandíbula/química , Maxilar/química , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Proteínas del Tejido Nervioso/análisis , Nestina/análisis , Análisis de Secuencia por Matrices de Oligonucleótidos , Osteogénesis/genética , Hueso Parietal/patología , Hueso Parietal/cirugía , Periostio/química , Periostio/trasplante , Proteínas de Unión al ARN/análisis , Distribución Aleatoria , Reacción en Cadena en Tiempo Real de la Polimerasa , Factores de Transcripción SOXE/análisis , Proteína Wnt1/análisis , Microtomografía por Rayos X/métodosRESUMEN
Treatment of large bone defects using synthetic scaffolds remain a challenge mainly due to insufficient vascularization. This study is to engineer a vascularized bone graft by integrating a vascularized biomimetic cell-sheet-engineered periosteum (CSEP) and a biodegradable macroporous beta-tricalcium phosphate (ß-TCP) scaffold. We first cultured human mesenchymal stem cells (hMSCs) to form cell sheet and human umbilical vascular endothelial cells (HUVECs) were then seeded on the undifferentiated hMSCs sheet to form vascularized cell sheet for mimicking the fibrous layer of native periosteum. A mineralized hMSCs sheet was cultured to mimic the cambium layer of native periosteum. This mineralized hMSCs sheet was first wrapped onto a cylindrical ß-TCP scaffold followed by wrapping the vascularized HUVEC/hMSC sheet, thus generating a biomimetic CSEP on the ß-TCP scaffold. A nonperiosteum structural cell sheets-covered ß-TCP and plain ß-TCP were used as controls. In vitro studies indicate that the undifferentiated hMSCs sheet facilitated HUVECs to form rich capillary-like networks. In vivo studies indicate that the biomimetic CSEP enhanced angiogenesis and functional anastomosis between the in vitro preformed human capillary networks and the mouse host vasculature. MicroCT analysis and osteocalcin staining show that the biomimetic CSEP/ß-TCP graft formed more bone matrix compared to the other groups. These results suggest that the CSEP that mimics the cellular components and spatial configuration of periosteum plays a critical role in vascularization and osteogenesis. Our studies suggest that a biomimetic periosteum-covered ß-TCP graft is a promising approach for bone regeneration.
Asunto(s)
Biomimética , Regeneración Ósea , Osteogénesis , Periostio/química , Ingeniería de Tejidos , Animales , Trasplante Óseo , Fosfatos de Calcio/química , Fosfatos de Calcio/uso terapéutico , Células Endoteliales de la Vena Umbilical Humana , Humanos , Células Madre Mesenquimatosas/química , Ratones , Osteocalcina/química , Andamios del Tejido/químicaRESUMEN
Inspection of patient-derived cells used in transplantation is non-invasive. Therefore, proteomics analysis using supernatants of cells cultured before transplantation is informative. In order to investigate the cell niche of bovine periosteal cells, supernatants of these cultured cells were subjected to 2-D electrophoresis followed by mass spectrometry, which identified type 1 collagen and the C-terminus of type 3 collagen. Only the C-terminal peptide from type 3 collagen was found in supernatants. It is known that type 3 collagen may be expressed intra- or extra-cellularly. Paraffin sections of the cultured cells were next examined by immunohistochemistry, which revealed that type 3 collagen regions besides the C-terminal peptide were present around the bovine periosteal cells but were not found in supernatants. Full-length type 3 collagen was closely associated with the cells, and only the C-terminal peptide was detectable in culture supernatants. Mass spectrometry analysis of partial peptide data combined with immunohistochemistry also indicated that uveal autoantigen with coiled coil domains and ankyrin repeats (UACA), exosome complex component RRP45 (EXOSC9), and thioredoxin-related transmembrane protein 2 (TMX2) were expressed in bovine periosteal cells. Results of this study indicate that analysis of culture supernatants before cell transplantation can provide useful biomarkers indicating the niche of cells used for transplantation.
Asunto(s)
Autoantígenos/metabolismo , Complejo Multienzimático de Ribonucleasas del Exosoma/metabolismo , Periostio/química , Periostio/metabolismo , Tiorredoxinas/metabolismo , Animales , Autoantígenos/química , Bovinos , Células Cultivadas , Electroforesis en Gel Bidimensional , Complejo Multienzimático de Ribonucleasas del Exosoma/química , Inmunohistoquímica , Espectrometría de Masas , Periostio/citología , Tiorredoxinas/químicaRESUMEN
A "sticker-like" PLGA nanosheet with microgrooved patterns is developed through a facile combination of spin coating and micropatterning techniques. The resulting microgrooved PLGA nanosheets can be physically adhered on flat or porous surfaces with excellent stability in aqueous environments and can harness the spatial arrangements of cells, which make it a promising candidate for generating biomimic periosteum for bone regenerative applications.
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Materiales Biomiméticos/química , Ácido Láctico/química , Nanoestructuras/química , Periostio/química , Ácido Poliglicólico/química , Animales , Huesos/química , Adhesión Celular , Células Cultivadas , Pollos , Humanos , Células Madre Mesenquimatosas/fisiología , Microscopía Electrónica de Rastreo , Osteogénesis , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Porosidad , Andamios del Tejido , Agua/químicaRESUMEN
Abnormal elongation of the styloid process, or Eagle syndrome, can be painful, and is associated with differential diagnoses including cranio-facial malformations and vasculo-neurological disturbances. The precise molecular mechanism leading to styloid process elongation is unknown. In this study, elongated styloid processes with periosteal fibrous ligament tissue were obtained from three patients with Eagle syndrome and examined by immunohistochemical methods using different antisera. In all cases, marked bony deposition was found at the apex of the styloid process. The osteogenetic proteins, such as osteonectin, osteocalcin, BMP-2, BMP-4, and RANKL were strongly positive by immunohistochemistry in both the ligament fibers and the periosteal membrane attached to the styloid process apex. Staining for protective proteins, HO-1, HSP-70, and HSP-90 was also positive. These results suggest that styloid process elongation is related to increased expression of osteogenetic and protective proteins. Therefore, we propose that Eagle syndrome results from a protective response to increased tensile stress in the ligament attached to the styloid process, which could also signal osteogenetic protein expression in the periosteal fibrous tissue.
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Osificación Heterotópica/fisiopatología , Osteogénesis/fisiología , Hueso Temporal/anomalías , Adulto , Fenómenos Biomecánicos , Proteína Morfogenética Ósea 2/análisis , Proteína Morfogenética Ósea 4/análisis , Proteínas HSP70 de Choque Térmico/análisis , Proteínas HSP90 de Choque Térmico/análisis , Hemo-Oxigenasa 1/análisis , Humanos , Inmunohistoquímica , Ligamentos/química , Ligamentos/fisiopatología , Masculino , Osificación Heterotópica/metabolismo , Osteocalcina/análisis , Osteonectina/análisis , Periostio/química , Periostio/fisiopatología , Ligando RANK/análisis , Estrés Mecánico , Hueso Temporal/química , Hueso Temporal/metabolismo , Hueso Temporal/fisiopatología , Adulto JovenRESUMEN
Cortical bone allografts suffer from high rates of failure due to poor integration with host tissue, leading to non-union, fracture, and infection following secondary procedures. Here, we report a method for modifying the surfaces of cortical bone with coatings that have biological functions that may help overcome these challenges. These chitosan-heparin coatings promote mesenchymal stem cell attachment and have significant antibacterial activity against both S. aureus and E. coli. Furthermore, their chemistry is similar to coatings we have reported on previously, which effectively stabilize and deliver heparin-binding growth factors. These coatings have potential as synthetic periosteum for improving bone allograft outcomes.