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1.
Int J Nanomedicine ; 16: 3509-3540, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34045855

RESUMO

The increasing incidence of bone-related disorders is causing a burden on the clinical scenario. Even though bone is one of the tissues that possess tremendous regenerative potential, certain bone anomalies need therapeutic intervention through appropriate delivery of a drug. Among several nanosystems and biologics that offer the potential to contribute towards bone healing, the exosomes from the class of extracellular vesicles are outstanding. Exosomes are extracellular nanovesicles that, apart from the various advantages, are standing out of the crowd for their ability to conduct cellular communication. The internal cargo of the exosomes is leading to its potential use in therapeutics. Exosomes are being unraveled in terms of the mechanism as well as application in targeting various diseases and tissues. Through this review, we have tried to understand and review all that is already established and the gap areas that still exist in utilizing them as drug delivery vehicles targeting the bone. The review highlights the potential of the exosomes towards their contribution to the drug delivery scenario in the bone microenvironment. A comparison of the pros and cons of exosomes with other prevalent drug delivery systems is also done. A section on the patents that have been generated so far from this field is included.


Assuntos
Osso e Ossos/citologia , Microambiente Celular/efeitos dos fármacos , Sistemas de Liberação de Medicamentos/métodos , Exossomos/metabolismo , Animais , Exossomos/efeitos dos fármacos , Humanos
2.
Int J Nanomedicine ; 16: 3161-3172, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33994785

RESUMO

Purpose: There is no definitive treatment for bisphosphonate-related osteonecrosis of the jaw (BRONJ). Small extracellular vesicles derived from adipose tissue (sEV-AT) have been proved efficient at promoting tissue regeneration. The aim of this study was to evaluate the effects of sEV-AT administration on BRONJ-like lesions in rats. Methods: Zoledronate (Zol) and dexamethasone (Dex) were subcutaneously administered to create a BRONJ rat model. Rats were randomly divided into three groups: 1) Control; 2) Zol+Dex; 3) sEV-AT. The maxillary left first molars were extracted two weeks after the first administration. In the sEV-AT group, sEV-AT were given intravenously every three days right after tooth extraction. We preformed occlusal view images, microcomputed tomography (µCT) and histological analysis to measure the regeneration of osseous and soft tissue in extraction sockets. Human umbilical vein endothelial cells (HUVECs) were isolated and cultured with endothelial cell medium (ECM). HUVECs were then divided into three groups: 1) Control: ECM; 2) Zol: ECM+Zol; 3) sEV-AT: ECM+Zol+sEV-AT. We evaluated the proliferation, tube formation and migration of HUVECs in each group. Results: Rats treated with Zol+Dex showed BRONJ-like lesions including open wounds, necrotic bones, empty osteocyte lacunae and reduced osteoclasts. sEV-AT administration reduced BRONJ-like lesions by promoting soft tissue healing. µCT results showed that bone volume in extraction sockets in the sEV-AT group was larger than the Zol+Dex group. Histological analysis showed less necrotic bones and empty osteocyte lacunae in the sEV-AT group compared to the Zol+Dex group. Histological analysis also showed more osteoclasts, collagen fibers and blood vessels in the sEV-AT group compared to the Zol+Dex group. Furthermore, sEV-AT enhanced the proliferation, migration and tube formation of HUVECs which were inhibited by Zol. Conclusion: Our findings indicate that sEV-AT prevent BRONJ in rats. Angiogenesis promotion contributes to the prevention of BRONJ.


Assuntos
Tecido Adiposo/citologia , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/prevenção & controle , Osso e Ossos/citologia , Vesículas Extracelulares/transplante , Neovascularização Fisiológica , Osteoclastos/citologia , Cicatrização , Animais , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/patologia , Proliferação de Células , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Dente Molar/patologia , Ratos , Ratos Sprague-Dawley , Extração Dentária
3.
Langmuir ; 37(14): 4137-4146, 2021 04 13.
Artigo em Inglês | MEDLINE | ID: mdl-33813823

RESUMO

Hydroxyapatite (HA) is the main inorganic component of human bones and teeth. It has good biocompatibility and bioactivity, which promotes its good application prospects in the field of bone drug carriers. In this study, tetraethylenepentamine-graphene (rGO-TEPA)/CaCO3:HA composite microspheres were prepared via microwave hydrothermal synthesis using rGO-TEPA/CaCO3 solid microspheres as intermediates. Furthermore, the incompletely transformed CaCO3 was removed by soaking in a citric acid buffer to obtain rGO-TEPA/HA hollow composite microspheres. The two types of as-prepared composite microspheres exhibited sea urchin-like structures, large BET surface areas, and good dispersibility. Mouse preosteoblast cells (MC3T3-E1) were used for in vitro cytotoxicity experiments. The in vitro cell viability test showed that the two composite drug carriers exhibited noncytotoxicity. Moreover, the doxorubicin (DOX) loading and releasing investigations revealed that the two types of prepared carriers had mild storage-release behaviors and good pH responsiveness. Hence, these rGO-TEPA/HA hollow microspheres have promising applications as bone drug carriers.


Assuntos
Materiais Biomiméticos , Osso e Ossos/metabolismo , Portadores de Fármacos/química , Portadores de Fármacos/metabolismo , Durapatita , Grafite , Microesferas , Ouriços-do-Mar , Animais , Osso e Ossos/citologia , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/farmacologia , Etilenodiaminas , Concentração de Íons de Hidrogênio , Camundongos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos
4.
Int J Mol Sci ; 22(8)2021 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-33924333

RESUMO

Bone formation starts near the end of the embryonic stage of development and continues throughout life during bone modeling and growth, remodeling, and when needed, regeneration. Bone-forming cells, traditionally termed osteoblasts, produce, assemble, and control the mineralization of the type I collagen-enriched bone matrix while participating in the regulation of other cell processes, such as osteoclastogenesis, and metabolic activities, such as phosphate homeostasis. Osteoblasts are generated by different cohorts of skeletal stem cells that arise from different embryonic specifications, which operate in the pre-natal and/or adult skeleton under the control of multiple regulators. In this review, we briefly define the cellular identity and function of osteoblasts and discuss the main populations of osteoprogenitor cells identified to date. We also provide examples of long-known and recently recognized regulatory pathways and mechanisms involved in the specification of the osteogenic lineage, as assessed by studies on mice models and human genetic skeletal diseases.


Assuntos
Osteoblastos/citologia , Osteogênese , Células-Tronco/citologia , Animais , Osso e Ossos/citologia , Osso e Ossos/embriologia , Epigênese Genética , Humanos , Osteogênese/genética , Transdução de Sinais
5.
Med Sci Monit ; 27: e929510, 2021 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-33828067

RESUMO

The directional differentiation of bone mesenchymal stem cells (BMSCs) is regulated by a variety of transcription factors and intracellular signaling pathways. In the past, it was thought that the directional differentiation of BMSCs was related to transforming growth factors, such as bone morphogenetic protein (BMP) and MAPK pathway. However, in recent years, some scholars have pointed out that the Wnt signaling pathway, which is a necessary complex network of protein interactions for biological growth and development, takes a significant role in this process and plays a major part in regulating the development of osteoblasts by exerting signal transduction into cells. Also, they have proved the Wnt protein therapeutic truly have positive effects on the viability and osteogenic capacity of bone graft. Recent studies have shown that microRNAs (miRNAs) play an important regulatory role in this process. MiRNAs such as miRNA-218, miRNA-335, miRNA-29, microRNA-30 and other miRNAs exert negative or positive effects on some crucial molecules in the Wnt/ß-catenin pathway, which in turn affect bone metabolism and osteopathy. Thus, miRNAs have been suggested as therapeutic targets for some metabolic bone diseases. This article aims to provide an update on the current status of microRNAs that target the Wnt signaling pathway in the regulation of osteogenesis and bone metabolism and includes a discussion of future areas of research, which can be a theoretical basis for bone metabolism-related diseases.


Assuntos
Doenças Ósseas/metabolismo , Osso e Ossos/metabolismo , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , Osteogênese/genética , Animais , Osso e Ossos/citologia , Humanos , Células-Tronco Mesenquimais/citologia , Via de Sinalização Wnt
6.
Phytomedicine ; 85: 153485, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33743412

RESUMO

BACKGROUND: Icariin (ICA) is a bioactive compound isolated from epimedium-derived flavonoids that modulates bone mesenchymal stem cell osteogenesis and adipogenesis. However, its precise mechanism in this process is unknown. PURPOSE: The purpose of this study was to elucidate the role of ICA on human bone mesenchymal stem cell (hBMSC) osteogenesis and adipogenesis by focusing on miR-23a mediated activation of the Wnt/ß-catenin signaling pathway. METHODS: After ICA treatment, hBMSC osteogenesis and adipogenesis were evaluated using alkaline phosphatase staining, an alkaline phosphatase activity assay, Oil Red O staining, and cellular triglyceride levels. Moreover, the mRNA and protein expression levels of osteogenic and adipogenic markers as well as key factors of the Wnt/ß-catenin signaling pathway were measured using quantitative reverse transcription polymerase chain reaction and western blotting. Lithium chloride, an activator of the Wnt/ß-catenin signaling pathway, was used as a positive control. Finally, to investigate the role of miR-23a in ICA-induced activation of the Wnt/ß-catenin signaling pathway, hBMSCs were transfected with miR-23a mimics or a miR-23a inhibitor. RESULTS: ICA significantly promoted hBMSC osteogenic differentiation by upregulating alkaline phosphatase activity and the expression of bone sialoprotein II (BSPII) and runt-related transcription factor-2 (Runx-2). In contrast, ICA inhibited hBMSC adipogenic differentiation by reducing lipid droplet formation and cellular triglyceride levels as well as by downregulating the expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) and CCAAT enhancer-binding protein-α (C/EBP-α). ICA mediated its effects on hBMSCs by activating the Wnt/ß-catenin signaling pathway. It did so by upregulating ß-catenin, low density lipoprotein receptor-related protein 5 (LRP5), and T cell factor 1 (TCF1). Notably, the up-regulation of these proteins was blocked by Dickkopf-related protein 1 (DKK1). Critically, the effects of ICA on hBMSCs were similar to that of the positive control, lithium chloride. Notably, ICA-induced activation of the Wnt/ß-catenin signaling pathway was significantly attenuated following miR-23a up-regulation. Conversely, miR-23a downregulation affected hBMSCs in the same manner as ICA; i.e., it activated the Wnt/ß-catenin signaling pathway. CONCLUSION: ICA promotes and inhibits, respectively, hBMSC osteogenesis and adipogenesis via miR-23a-mediated activation of the Wnt/ß-catenin signaling pathway.


Assuntos
Adipogenia/efeitos dos fármacos , Flavonoides/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , MicroRNAs/genética , Osteogênese/efeitos dos fármacos , Via de Sinalização Wnt , Osso e Ossos/citologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Epimedium/química , Humanos , Sialoproteína de Ligação à Integrina/metabolismo , Células-Tronco Mesenquimais/citologia , beta Catenina/metabolismo
7.
J Mater Chem B ; 9(8): 2033-2041, 2021 03 04.
Artigo em Inglês | MEDLINE | ID: mdl-33587079

RESUMO

Xenogeneic bones are potential templates for bone regeneration. In this study, decellularized porcine bone powder with attenuated immunogenicity was incorporated into a photocurable hydrogel, gelatin methacryloyl (GelMA), to obtain scaffolds with good mechanical properties for bone tissue engineering. The decellularized bone powder (DCB)-GelMA hybrid scaffolds had higher compressive strength and stiffness values when the DCB content was increased. In vitro evaluations revealed the biocompatibility of these scaffolds. The scaffolds could induce human bone marrow mesenchymal stem cells (hMSCs) to undergo osteogenic differentiation even in the absence of an induction medium. The efficiency of the scaffolds for bone regeneration applications was further evaluated using an in vivo cranial bone defect model in rats. Micro-CT images showed that the hybrid scaffolds with 20% DCB content had the best effect in promoting new bone regeneration. Thus, it was concluded that the DCB-GelMA hybrid scaffolds have high potential in bone tissue engineering applications.


Assuntos
Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/fisiologia , Hidrogéis/química , Engenharia Tecidual , Tecidos Suporte/química , Animais , Osso e Ossos/citologia , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Força Compressiva , Gelatina/química , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Metacrilatos/química , Osteogênese/efeitos dos fármacos , Suínos
8.
FASEB J ; 35(3): e21424, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33609323

RESUMO

Gorham-Stout disease (GSD) is a very rare disease characterized by increased bone erosion with angiomatous proliferation. The mechanisms underlying this disorder have not been deeply investigated. Due to its rarity, no guidelines are currently available for treatment and management of GSD. We recently evaluated the cellular alterations of the bone remodeling in patients showing that osteoclast precursors displayed increased ability to differentiate into osteoclasts and that affected osteoclasts resorb bone more actively than control cells. Moreover, osteoblasts isolated from a patient showed a defective ability to form mineralized nodules. In this paper, we investigated the molecular pathways involved in the cellular defects of GSD bone cells. For this study, we recruited nine patients and performed miRNome analysis of bone cells. Between the 178 miRNAs robustly expressed in GSD osteoclasts, significant modulation of three miRNAs (miR-1246, miR-1-3p, and miR-137-3p) involved in the regulation of osteoclast formation and activity or in the angiomatous proliferation was found in patients' cells. Interestingly, miR-1246 was also up-regulated in serum exosomes from patients. Analysis of miRNAs from patient osteoblasts suggested alteration of miR-204a-5p, miR-615-3p and miR-378a-3p regulating osteoblast function and differentiation. The resulting miRNA pattern may help to understand better the mechanisms involved in GSD and to identify new potential therapeutic targets for this rare disease.


Assuntos
Osso e Ossos/citologia , MicroRNAs/genética , Osteoclastos/metabolismo , Osteócitos/metabolismo , Osteólise Essencial/sangue , Adolescente , Osso e Ossos/metabolismo , Diferenciação Celular/genética , Criança , Exossomos/metabolismo , Feminino , Humanos , Masculino , MicroRNAs/sangue , Osteólise Essencial/fisiopatologia
9.
J Mater Chem B ; 9(10): 2394-2406, 2021 03 17.
Artigo em Inglês | MEDLINE | ID: mdl-33625433

RESUMO

Although clay-based nanocomposite hydrogels have been widely explored, their instability in hot water and saline solution inhibits their applications in biomedical engineering, and the exploration of clay-based nanocomposite hydrogels in bone defect repair is even less. In this work, we developed a stable clay-based nanocomposite hydrogel using 4-acryloylmorpholine as the monomer. After UV light illumination, the obtained poly(4-acryloylmorpholine) clay-based nanocomposite hydrogel (poly(4-acry)-clay nanocomposite hydrogel) exhibits excellent mechanical properties due to the hydrogen bond interactions between the poly(4-acryloylmorpholine) chains and the physical crosslinking effect of the nanoclay. Besides good biocompatibility, the sustainable release of intrinsic Mg2+ and Si4+ from the poly(4-acry)-clay nanocomposite hydrogel endows the system with excellent ability to promote the osteogenic differentiation of primary rat osteoblasts (ROBs) and can promote new bone formation effectively after implantation. We anticipate that these kinds of clay-based nanocomposite hydrogels with sustained release of bioactive ions will open a new avenue for the development of novel biomaterials for bone regeneration.


Assuntos
Osso e Ossos/efeitos dos fármacos , Argila/química , Hidrogéis/química , Hidrogéis/farmacologia , Fenômenos Mecânicos , Nanocompostos/química , Acrilamidas/química , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/citologia , Osso e Ossos/fisiologia , Diferenciação Celular/efeitos dos fármacos , Morfolinas/química , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Ratos
10.
Molecules ; 26(4)2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33572384

RESUMO

Advances in vibrational spectroscopy have propelled new insights into the molecular composition and structure of biological tissues. In this review, we discuss common modalities and techniques of vibrational spectroscopy, and present key examples to illustrate how they have been applied to enrich the assessment of connective tissues. In particular, we focus on applications of Fourier transform infrared (FTIR), near infrared (NIR) and Raman spectroscopy to assess cartilage and bone properties. We present strengths and limitations of each approach and discuss how the combination of spectrometers with microscopes (hyperspectral imaging) and fiber optic probes have greatly advanced their biomedical applications. We show how these modalities may be used to evaluate virtually any type of sample (ex vivo, in situ or in vivo) and how "spectral fingerprints" can be interpreted to quantify outcomes related to tissue composition and quality. We highlight the unparalleled advantage of vibrational spectroscopy as a label-free and often nondestructive approach to assess properties of the extracellular matrix (ECM) associated with normal, developing, aging, pathological and treated tissues. We believe this review will assist readers not only in better understanding applications of FTIR, NIR and Raman spectroscopy, but also in implementing these approaches for their own research projects.


Assuntos
Osso e Ossos/citologia , Cartilagem/citologia , Tecido Conjuntivo/fisiologia , Análise Espectral Raman/métodos , Animais , Osso e Ossos/química , Cartilagem/química , Tecnologia de Fibra Óptica , Humanos , Espectroscopia de Infravermelho com Transformada de Fourier , Espectroscopia de Luz Próxima ao Infravermelho
11.
Int J Biol Macromol ; 175: 379-395, 2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33556401

RESUMO

Biomaterials function as an essential aspect of tissue engineering and have a profound impact on cell growth and subsequent tissue regeneration. The development of new biomaterials requires a potential platform to understand the host-biomaterial interaction, which is crucial for successful biomaterial implantation. Biomaterials analyzed in rodent models for in vivo research are cost-effective but tedious, and the practice has many technical difficulties. As an alternative, zebrafish provide an excellent biomaterial testing platform over the current rodent models. During growth and recovery, zebrafish bone morphogenesis shows a variety of inductive signals involved in the cycle that are close to those influencing differentiation of bone and cartilage in mammals, including humans. This platform is cheap, optically transparent, quick to change genes, and provides reliable reproducibility on short life cycles. Chitosan is a well-known biomaterial in the field of tissue engineering. In view of its documented use in bone regeneration, the biological characterization of chitosan-based bioactive materials in the zebrafish model has been featured in an outstanding note. We, therefore, outlined this review of the zebrafish as a potential in vivo research model for the rapid characterization of the biological properties of new biomaterials for bone tissue engineering applications.


Assuntos
Osso e Ossos/metabolismo , Quitosana/química , Engenharia Tecidual/métodos , Animais , Materiais Biocompatíveis , Regeneração Óssea/fisiologia , Osso e Ossos/citologia , Cartilagem/citologia , Quitosana/metabolismo , Teste de Materiais , Modelos Animais , Reprodutibilidade dos Testes , Tecidos Suporte , Peixe-Zebra/metabolismo
12.
Nat Commun ; 12(1): 1031, 2021 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-33589620

RESUMO

The application of physical stimuli to cell cultures has shown potential to modulate multiple cellular functions including migration, differentiation and survival. However, the relevance of these in vitro models to future potential extrapolation in vivo depends on whether stimuli can be applied "externally", without invasive procedures. Here, we report on the fabrication and exploitation of dynamic additive-manufactured Janus scaffolds that are activated on-command via external application of ultrasounds, resulting in a mechanical nanovibration that is transmitted to the surrounding cells. Janus scaffolds were spontaneously formed via phase-segregation of biodegradable polycaprolactone (PCL) and polylactide (PLA) blends during the manufacturing process and behave as ultrasound transducers (acoustic to mechanical) where the PLA and PCL phases represent the active and backing materials, respectively. Remote stimulation of Janus scaffolds led to enhanced cell proliferation, matrix deposition and osteogenic differentiation of seeded human bone marrow derived stromal cells (hBMSCs) via formation and activation of voltage-gated calcium ion channels.


Assuntos
Plásticos Biodegradáveis/farmacologia , Mecanotransdução Celular , Células-Tronco Mesenquimais/efeitos dos fármacos , Poliésteres/farmacologia , Tecidos Suporte , Plásticos Biodegradáveis/química , Regeneração Óssea/genética , Osso e Ossos/citologia , Osso e Ossos/metabolismo , Canais de Cálcio Ativados pela Liberação de Cálcio/fisiologia , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Poliésteres/química , Impressão Tridimensional , Engenharia Tecidual/métodos , Ondas Ultrassônicas
13.
Molecules ; 26(4)2021 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-33562111

RESUMO

Microbial polyhydroxyalkanoates (PHA) are proteinaceous storage granules ranging from 100 nm to 500 nm. Bacillus sp. serve as unique bioplastic sources of short-chain length and medium-chain length PHA showcasing properties such as biodegradability, thermostability, and appreciable mechanical strength. The PHA can be enhanced by adding functional groups to make it a more industrially useful biomaterial. PHA blends with hydroxyapatite to form nanocomposites with desirable features of compressibility. The reinforced matrices result in nanocomposites that possess significantly improved mechanical and thermal properties both in solid and melt states along with enhanced gas barrier properties compared to conventional filler composites. These superior qualities extend the polymeric composites' applications to aggressive environments where the neat polymers are likely to fail. This nanocomposite can be used in different industries as nanofillers, drug carriers for packaging essential hormones and microcapsules, etc. For fabricating a bone scaffold, electrospun nanofibrils made from biocomposite of hydroxyapatite and polyhydroxy butyrate, a form of PHA, can be incorporated with the targeted tissue. The other methods for making a polymer scaffold, includes gas foaming, lyophilization, sol-gel, and solvent casting method. In this review, PHA as a sustainable eco-friendly NextGen biomaterial from bacterial sources especially Bacillus cereus, and its application for fabricating bone scaffold using different strategies for bone regeneration have been discussed.


Assuntos
Bacillus/metabolismo , Biopolímeros/metabolismo , Osso e Ossos/efeitos dos fármacos , Poli-Hidroxialcanoatos/biossíntese , Poli-Hidroxialcanoatos/farmacologia , Tecidos Suporte/química , Animais , Osso e Ossos/citologia , Osso e Ossos/fisiologia , Humanos , Poli-Hidroxialcanoatos/química
14.
Int J Mol Sci ; 22(3)2021 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-33572704

RESUMO

Notch1-4 receptors and their signaling pathways are expressed in almost all organ systems and play a pivotal role in cell fate decision by coordinating cell proliferation, differentiation and apoptosis. Differential expression and activation of Notch signaling pathways has been observed in a variety of organs and tissues under physiological and pathological conditions. Bone tissue represents a dynamic system, which is constantly remodeled throughout life. In bone, Notch receptors have been shown to control remodeling and regeneration. Numerous functions have been assigned to Notch receptors and ligands, including osteoblast differentiation and matrix mineralization, osteoclast recruitment and cell fusion and osteoblast/osteoclast progenitor cell proliferation. The expression and function of Notch1-4 in the skeleton are distinct and closely depend on the temporal expression at different differentiation stages. This review addresses the current knowledge on Notch signaling in adult bone with emphasis on metabolism, bone regeneration and degenerative skeletal disorders, as well as congenital disorders associated with mutant Notch genes. Moreover, the crosstalk between Notch signaling and other important pathways involved in bone turnover, including Wnt/ß-catenin, BMP and RANKL/OPG, are outlined.


Assuntos
Regeneração Óssea , Osso e Ossos/metabolismo , Receptores Notch/metabolismo , Transdução de Sinais , Animais , Osso e Ossos/citologia , Humanos , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteoclastos/citologia , Osteoclastos/metabolismo , Osteócitos/citologia , Osteócitos/metabolismo
15.
Methods Mol Biol ; 2286: 95-105, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33534112

RESUMO

Bone marrow stromal cells (BMSCs, also known as bone marrow mesenchymal stem cells) are a plastic-adherent heterogeneous cell population that contain inherent skeletal progenitors and a subset of multipotential skeletal stem cells (SSCs). Application of BMSCs in therapeutic protocols implies its isolation and expansion under good manufacturing practices (GMP). Here we describe the procedures we have found to successfully generate practical BMSCs numbers, with preserved biological potency.


Assuntos
Tecnologia Biomédica/normas , Células da Medula Óssea/citologia , Osso e Ossos/citologia , Cultura Primária de Células/métodos , Antígenos CD34/genética , Antígenos CD34/metabolismo , Tecnologia Biomédica/métodos , Células Cultivadas , Técnicas de Cocultura/economia , Técnicas de Cocultura/métodos , Técnicas de Cocultura/normas , Custos e Análise de Custo , Meios de Cultura Livres de Soro/química , Humanos , Guias de Prática Clínica como Assunto , Cultura Primária de Células/economia , Cultura Primária de Células/normas , Células Estromais/citologia , Células Estromais/metabolismo
16.
Int J Mol Sci ; 22(2)2021 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-33466904

RESUMO

Reconstruction of segmental bone defects by autologous bone grafting is still the standard of care but presents challenges including anatomical availability and potential donor site morbidity. The process of 3D bioprinting, the application of 3D printing for direct fabrication of living tissue, opens new possibilities for highly personalized tissue implants, making it an appealing alternative to autologous bone grafts. One of the most crucial hurdles for the clinical application of 3D bioprinting is the choice of a suitable cell source, which should be minimally invasive, with high osteogenic potential, with fast, easy expansion. In this study, mesenchymal progenitor cells were isolated from clinically relevant human bone biopsy sites (explant cultures from alveolar bone, iliac crest and fibula; bone marrow aspirates; and periosteal bone shaving from the mastoid) and 3D bioprinted using projection-based stereolithography. Printed constructs were cultivated for 28 days and analyzed regarding their osteogenic potential by assessing viability, mineralization, and gene expression. While viability levels of all cell sources were comparable over the course of the cultivation, cells obtained by periosteal bone shaving showed higher mineralization of the print matrix, with gene expression data suggesting advanced osteogenic differentiation. These results indicate that periosteum-derived cells represent a highly promising cell source for translational bioprinting of bone tissue given their superior osteogenic potential as well as their minimally invasive obtainability.


Assuntos
Células da Medula Óssea/metabolismo , Transplante Ósseo/métodos , Osso e Ossos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Biossíntese de Proteínas , Engenharia Tecidual/métodos , Adulto , Bioimpressão/métodos , Células da Medula Óssea/citologia , Osso e Ossos/citologia , Diferenciação Celular/genética , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/citologia , Osteogênese/genética , Impressão Tridimensional , Tecidos Suporte , Transplante Autólogo
17.
Int J Mol Sci ; 22(2)2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33467447

RESUMO

Mesenchymal stem cells (MSCs) are the main cell players in tissue repair and thanks to their self-renewal and multi-lineage differentiation capabilities, they gained significant attention as cell source for tissue engineering (TE) approaches aimed at restoring bone and cartilage defects. Despite significant progress, their therapeutic application remains debated: the TE construct often fails to completely restore the biomechanical properties of the native tissue, leading to poor clinical outcomes in the long term. Pulsed electromagnetic fields (PEMFs) are currently used as a safe and non-invasive treatment to enhance bone healing and to provide joint protection. PEMFs enhance both osteogenic and chondrogenic differentiation of MSCs. Here, we provide extensive review of the signaling pathways modulated by PEMFs during MSCs osteogenic and chondrogenic differentiation. Particular attention has been given to the PEMF-mediated activation of the adenosine signaling and their regulation of the inflammatory response as key player in TE approaches. Overall, the application of PEMFs in tissue repair is foreseen: (1) in vitro: to improve the functional and mechanical properties of the engineered construct; (2) in vivo: (i) to favor graft integration, (ii) to control the local inflammatory response, and (iii) to foster tissue repair from both implanted and resident MSCs cells.


Assuntos
Diferenciação Celular/fisiologia , Condrogênese/fisiologia , Campos Eletromagnéticos , Células-Tronco Mesenquimais/citologia , Osteogênese/fisiologia , Transdução de Sinais/fisiologia , Osso e Ossos/citologia , Células Cultivadas , Humanos , Engenharia Tecidual/métodos
18.
Nat Metab ; 3(1): 11-20, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33398192

RESUMO

The skeleton is diverse in its functions, which include mechanical support, movement, blood cell production, mineral storage and endocrine regulation. This multifaceted role is achieved through an interplay of osteoblasts, chondrocytes, bone marrow adipocytes and stromal cells, all generated from skeletal stem cells. Emerging evidence shows the importance of cellular metabolism in the molecular control of the skeletal system. The different skeletal cell types not only have distinct metabolic demands relating to their particular functions but also are affected by microenvironmental constraints. Specific metabolites control skeletal stem cell maintenance, direct lineage allocation and mediate cellular communication. Here, we discuss recent findings on the roles of cellular metabolism in determining skeletal stem cell fate, coordinating osteoblast and chondrocyte function, and organizing stromal support of haematopoiesis. We also consider metabolic dysregulation in skeletal ageing and degenerative diseases, and provide an outlook on how the field may evolve in the coming years.


Assuntos
Doenças Ósseas/fisiopatologia , Osso e Ossos/citologia , Animais , Células da Medula Óssea , Osso e Ossos/fisiologia , Osso e Ossos/fisiopatologia , Comunicação Celular , Linhagem da Célula , Senescência Celular , Humanos , Osteoblastos/metabolismo , Células-Tronco/metabolismo
19.
Nat Mater ; 20(1): 108-118, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32958876

RESUMO

The maintenance of human skeletal stem cells (hSSCs) and their progeny in bone defects is a major challenge. Here, we report on a transplantable bandage containing a three-dimensional Wnt-induced osteogenic tissue model (WIOTM). This bandage facilitates the long-term viability of hSSCs (8 weeks) and their progeny, and enables bone repair in an in vivo mouse model of critical-sized calvarial defects. The newly forming bone is structurally comparable to mature cortical bone and consists of human and murine cells. Furthermore, we show that the mechanism of WIOTM formation is governed by Wnt-mediated asymmetric cell division of hSSCs. Covalently immobilizing Wnts onto synthetic materials can polarize single dividing hSSCs, orient the spindle and simultaneously generate a Wnt-proximal hSSC and a differentiation-prone Wnt-distal cell. Our results provide insight into the regulation of human osteogenesis and represent a promising approach to deliver human osteogenic constructs that can survive in vivo and contribute to bone repair.


Assuntos
Osso e Ossos/citologia , Divisão Celular , Osteogênese , Crânio/citologia , Células-Tronco/citologia , Engenharia Tecidual/métodos , Proteínas Wnt/metabolismo , Animais , Humanos , Camundongos , Crânio/fisiologia
20.
Life Sci ; 264: 118502, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33031825

RESUMO

Bone tissue engineering compasses the use of mesenchymal stem cells (MSCs) along with engineered biomaterial construct to augment bone regeneration. Till now, MSCs were isolated from various sources and used in cellular constructs. For the first time, in this study, MSCs were isolated from human Ovarian Follicular Fluid (OFF) and characterized by CD 44+ and CD 105+ markers via confocal microscopy and flow cytometry. Additionally, MSCs stemness, proliferation and colony-forming unit ability, multi-lineage differentiation potential were also studied. To test its suitability for bone tissue engineering applications, we grew the MSCs with the conditioned medium obtained from biocomposite scaffold by fusing a natural polymer, Chitosan (CS) and a synthetic polymer, Polycaprolactone (PCL) and the scaffold were coated with Zinc divalent ions to impart osteogenic properties. The physico-chemical characterization of scaffold, such as FTIR, XRD, and SEM studies was carried out. The biological characterization showed that the scaffolds were compatible with MSCs and promoted osteoblast differentiation which was confirmed at both cellular and molecular levels. The cellular construct increased calcium deposition, analyzed by alizarin red staining and ALP activity at cellular level. At the molecular level, the osteoblast markers expression such as Runx2 and type 1 collagen mRNAs, and osteonectin (ON) and osteocalcin (OC) secretory proteins were increased in the presence of scaffold. Overall, the current study recommends that MSCs can be easily obtained from human waste OFF, and grown in standard in vitro conditions. Successful growth of such MSCs with CS/PCL/Zn scaffold opens new avenues in utilizing the cell source for bone tissue engineering.


Assuntos
Materiais Biocompatíveis , Regeneração Óssea/fisiologia , Líquido Folicular/fisiologia , Folículo Ovariano/fisiologia , Engenharia Tecidual/métodos , Tecidos Suporte , Adulto , Materiais Biocompatíveis/administração & dosagem , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/citologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/fisiologia , Células Cultivadas , Quitosana/administração & dosagem , Feminino , Líquido Folicular/citologia , Líquido Folicular/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais , Recuperação de Oócitos/métodos , Osteogênese/efeitos dos fármacos , Osteogênese/fisiologia , Folículo Ovariano/efeitos dos fármacos , Poliésteres/administração & dosagem , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Difração de Raios X/métodos , Zinco/administração & dosagem
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