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1.
Tissue Cell ; 77: 101849, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35728334

RESUMO

Copper (Cu) ions have been found to exert antibacterial and angiogenic effects. However, some studies have indicated that it inhibits osteogenesis at high concentrations. On the other hand, L-arginine (Arg) is a semi-essential amino acid required for various biological processes, including osteogenic and angiogenic activities. As a result, we hypothesized that combining Arg with Cu ions would reduce its inhibitory effects on osteogenesis while increasing its angiogenic and antibacterial capabilities. To assess osteogenic and angiogenic activities, we employed rat bone marrow mesenchymal stem cells (MSCs) and human umbilical vein endothelial cells (HUVECs), respectively. The gram-positive bacteria Staphylococcus epidermidis (S. epidermidis), Staphylococcus aureus (S. aureus), and the gram-negative bacterium Escherichia coli (E. coli) were used to investigate bacterial behaviors. According to ALP activity and calcium deposition outcomes, copper ions inhibited osteogenic development of MSCs at 100 µM; however, Arg supplementation somewhat mitigated the inhibitory effects. Furthermore, Copper and Arg synergistically stimulated migration and tube formation of HUVECs. According to our findings, copper ions and Arg in the range of 1-100 µM had no antibacterial effect on any examined bacteria. However, at a dose of 20 mM, copper demonstrated antibacterial activity, which was boosted by Arg. Overall, these findings suggest that a combination of copper and Arg may be more beneficial for bone regeneration than either copper or Arg alone.

2.
Tissue Eng Regen Med ; 19(4): 839-852, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35199306

RESUMO

BACKGROUND: In the aftermath of bone injuries, such as cranium and sternum, bone wax (BW) is used to control bleeding from the bone surfaces during surgery. Made up of artificial substances, however, it is associated with many complications such as inflammation, increased risk for infection, and bone repair delay. We, therefore, in this study set out to design and evaluate a novel BW without the above-mentioned side-effects reported for other therapies. METHODS: The pastes (new BW(s)) were prepared in the laboratory and examined by MTT, MIC, MBC, and degradability tests. Then, 60 adult male Wistar rats, divided into six equal groups including chitosan (CT), CT-octacalcium phosphate (OCP), CT-periostin (Post), CT-OCP-Post, Control (Ctrl), and BW, underwent sternotomy surgery. Once the surgeries were completed, the bone repair was assessed radiologically and thereafter clinically in vivo and in vitro using CT-scan, H&E, ELISA, and qRT-PCR. RESULTS: All pastes displayed antibacterial properties and the CT-Post group had the highest cell viability compared to the control group. In contrast to the BW, CT-Post group demonstrated weight changes in the degradability test. In the CT-Post group, more number of osteocyte cells, high trabeculae percentage, and the least fibrous connective tissue were observed compared to other groups. Additionally, in comparison to the CT and Ctrl groups, higher alkaline phosphatase activity, as well as decreased level of serum tumor necrosis factor-α, interleukin-6, and OCN in the CT-Post group was evident. Finally, Runx2, OPG, and RANKL genes' expression was significantly higher in the CT-Post group than in other groups. CONCLUSION: Our results provide insights into the desirability of pastes in terms of cellular viability, degradability, antibacterial properties, and surgical site restoration compared to the BW group. Besides, Periostin could enhance the osteogenic properties of bone tissue defect site.


Assuntos
Materiais Biocompatíveis , Moléculas de Adesão Celular , Quitosana , Esterno , Animais , Antibacterianos , Moléculas de Adesão Celular/administração & dosagem , Quitosana/farmacologia , Interleucina-6/sangue , Masculino , Ratos , Ratos Wistar , Esternotomia , Esterno/cirurgia , Tecidos Suporte , Fator de Necrose Tumoral alfa/sangue
3.
Asian Pac J Cancer Prev ; 23(2): 731-741, 2022 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-35225487

RESUMO

BACKGROUND: Breast Cancer (BC) is a malignancy with high mortality among women. Recently, scaffold-based three-dimensional (3D) models have been developed for anti-cancer drug research. The present study aimed to investigate the anti-proliferative effects of Astragalus hamosus (A. hamosus) in 3D fibrin gel against MCF-7 cell line. We have also evaluated anti-proliferative effect of A. hamosus differences between 3D and 2D cultures. METHODS: The fibrin gel formulation was first optimized by testing the structural and mechanical properties. Then the cytotoxic effect of A. hamosus extract was assessed on MCF-7 cells by MTT assay. Cell apoptosis was evaluated using TUNEL method and flow cytometry. Cell cycle and proliferation were analyzed by flow cytometry. Apoptosis-related gene expression such as Bcl-2, caspase-3, -8 and -9 were quantified by real time-PCR. RESULTS: TUNEL staining showed a significant damage accompanied with cell apoptosis. Flow cytometry analysis revealed that apoptosis increased after treatment with A. hamosus extract in 3D culture model compared to 2D culture. The A. hamosus extract arrested cell cycle in the S and G2/M phases in 3D model while in the 2D culture G0/G1 phase was affected. Treatment with A. hamosus extract led to upregulation of the caspase-3, -8 and -9 genes and downregulation of the Ki-67 in the 3D-culture compared with the 2D culture. CONCLUSION: These results indicated that A. hamosus extract could be used as a therapeutic candidate for BC due to its anti-proliferative effects. Furthermore, 3D fibrin gel could be better than 2D-cultured cells in simulating important tumor characteristics in vivo, namely, anti-proliferative and anti-apoptotic features.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Astrágalo (Planta)/química , Neoplasias da Mama/tratamento farmacológico , Técnicas de Cultura de Células em Três Dimensões/métodos , Extratos Vegetais/farmacologia , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Géis , Humanos , Células MCF-7
4.
Biomed Res Int ; 2022: 7638245, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35118158

RESUMO

Distinctive characteristics of articular cartilage such as avascularity and low chondrocyte conversion rate present numerous challenges for orthopedists. Tissue engineering is a novel approach that ameliorates the regeneration process by exploiting the potential of cells, biodegradable materials, and growth factors. However, problems exist with the use of tissue-engineered construct, the most important of which is scaffold-cartilage integration. Recently, many attempts have been made to address this challenge via manipulation of cellular, material, and biomolecular composition of engineered tissue. Hence, in this review, we highlight strategies that facilitate cartilage-scaffold integration. Recent advances in where efficient integration between a scaffold and native cartilage could be achieved are emphasized, in addition to the positive aspects and remaining problems that will drive future research.


Assuntos
Cartilagem Articular , Engenharia Tecidual , Condrócitos , Regeneração , Tecidos Suporte
5.
J Biomater Sci Polym Ed ; 33(5): 605-626, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-34844507

RESUMO

Polymeric and tetracalcium phosphate (TTCP)-containing polymeric scaffolds were fabricated using a freeze-drying technique, with a homogenous solution of hydroxyethyl cellulose (HEC)/hyaluronic acid (HA)/gelatin (G) or suspension of 15 or 20% TTCP) particles in HEC/HA/G solution. The morphology, phase composition, chemical bands, and swelling behavior of the scaffold were determined. In vitro fibroblast cell viability and migration potential of the scaffolds were determined by MTT, live/dead staining, and scratch assay for wound healing. The in vivo chick embryo angiogenesis test was also carried out. Finally, the initial antibacterial activity of the scaffolds was determined using Staphylococcus aureus. The scaffolds exhibited an enormous porous structure in which the size of pores increased by the presence of TTCP particles. While the polymeric scaffold was amorphous, the formation of low crystalline hydroxyapatite phase and the initial TTCP particles was determined in the composition of TTCP-added scaffolds. TTCP increased swelling behavior of the polymeric scaffold in PBS. The results demonstrated that the amount of TTCP was a crucial factor in cell life. A high concentration of TTCP could restrict cell viability, although all the scaffolds were nontoxic. The scratch assessments determined better cell migration and wound closure in treating with TTCP-containing scaffolds so that after 24 h, a wound closure of 100% was observed. Furthermore, TTCP-incorporated scaffolds significantly improved the angiogenesis, in the chick embryo test. The presence of TTCP had a significant effect on reducing the bacterial activity and 20% TTCP-containing scaffold exhibited better antibacterial activity than the others.


Assuntos
Gelatina , Ácido Hialurônico , Animais , Antibacterianos/farmacologia , Fosfatos de Cálcio , Celulose , Embrião de Galinha , Gelatina/química , Ácido Hialurônico/química , Tecidos Suporte/química
6.
Biomed Pharmacother ; 146: 112584, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34968921

RESUMO

Heart muscle injury and an elevated troponin level signify myocardial infarction (MI), which may result in defective and uncoordinated segments, reduced cardiac output, and ultimately, death. Physicians apply thrombolytic therapy, coronary artery bypass graft (CABG) surgery, or percutaneous coronary intervention (PCI) to recanalize and restore blood flow to the coronary arteries, albeit they were not convincingly able to solve the heart problems. Thus, researchers aim to introduce novel substitutional therapies for regenerating and functionalizing damaged cardiac tissue based on engineering concepts. Cell-based engineering approaches, utilizing biomaterials, gene, drug, growth factor delivery systems, and tissue engineering are the most leading studies in the field of heart regeneration. Also, understanding the primary cause of MI and thus selecting the most efficient treatment method can be enhanced by preparing microdevices so-called heart-on-a-chip. In this regard, microfluidic approaches can be used as diagnostic platforms or drug screening in cardiac disease treatment. Additionally, bioprinting technique with whole organ 3D printing of human heart with major vessels, cardiomyocytes and endothelial cells can be an ideal goal for cardiac tissue engineering and remarkable achievement in near future. Consequently, this review discusses the different aspects, advancements, and challenges of the mentioned methods with presenting the advantages and disadvantages, chronological indications, and application prospects of various novel therapeutic approaches.


Assuntos
Infarto do Miocárdio/fisiopatologia , Regeneração/fisiologia , Materiais Biocompatíveis/metabolismo , Engenharia Celular/métodos , Sistemas de Liberação de Medicamentos/métodos , Vesículas Extracelulares/metabolismo , Fibroblastos/metabolismo , Terapia Genética/métodos , Insuficiência Cardíaca/fisiopatologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Células-Tronco Mesenquimais/metabolismo , Microfluídica/métodos , Mioblastos Esqueléticos/metabolismo , Isquemia Miocárdica/fisiopatologia , Células-Tronco/metabolismo , Engenharia Tecidual/métodos
7.
Biomed Pharmacother ; 146: 112529, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34906773

RESUMO

Spinal cord injury (SCI) is a central nervous system (CNS) devastate event that is commonly caused by traumatic or non-traumatic events. The reinnervation of spinal cord axons is hampered through a myriad of devices counting on the damaged myelin, inflammation, glial scar, and defective inhibitory molecules. Unfortunately, an effective treatment to completely repair SCI and improve functional recovery has not been found. In this regard, strategies such as using cells, biomaterials, biomolecules, and drugs have been reported to be effective for SCI recovery. Furthermore, recent advances in combinatorial treatments, which address various aspects of SCI pathophysiology, provide optimistic outcomes for spinal cord regeneration. According to the global importance of SCI, the goal of this article review is to provide an overview of the pathophysiology of SCI, with an emphasis on the latest modes of intervention and current advanced approaches for the treatment of SCI, in conjunction with an assessment of combinatorial approaches in preclinical and clinical trials. So, this article can give scientists and clinicians' clues to help them better understand how to construct preclinical and clinical studies that could lead to a breakthrough in spinal cord regeneration.


Assuntos
Traumatismos da Medula Espinal , Engenharia Tecidual , Materiais Biocompatíveis/uso terapêutico , Terapia Baseada em Transplante de Células e Tecidos , Humanos , Medula Espinal , Traumatismos da Medula Espinal/tratamento farmacológico
8.
Regen Med ; 16(8): 719-731, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34287065

RESUMO

Aim: To fabricate mature cartilage for implantation, developmental biological processes and proteins should be understood and employed. Methods: A systems biology study of all protein-coding genes participating in cartilage regeneration resulted in a network graph with 11 nodes and 28 edges. Gene ontology and centrality analysis were performed based on the degree index. Results: The four most crucial biological processes along with the seven most interactive proteins involved in cartilage regeneration were identified. Some proteins, which are under serious discussion in cartilage developmental and disease processes, are included in regeneration. Conclusions: Findings positively correlate with the literature, supporting the use of the four most impressive proteins as growth factors applicable to cartilage tissue engineering, including COL2A1, SOX9, CTGF and TGFß1.


Assuntos
Cartilagem Articular , Condrócitos , Cartilagem , Peptídeos e Proteínas de Sinalização Intercelular , Regeneração , Engenharia Tecidual
9.
Tissue Cell ; 72: 101553, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33975231

RESUMO

The principal purpose of tissue engineering is to stimulate the injured or unhealthy tissues to revive their primary function through the simultaneous use of chemical agents, cells, and biocompatible materials. Still, choosing the appropriate protein as a growth factor (GF) for tissue engineering is vital to fabricate artificial tissues and accelerate the regeneration procedure. In this study, the angiogenesis and osteogenesis-related proteins' interactions are studied using their related network. Three major biological processes, including osteogenesis, angiogenesis, and angiogenesis regulation, were investigated by creating a protein-protein interaction (PPI) network (45 nodes and 237 edges) of bone regeneration efficient proteins. Furthermore, a gene ontology and a centrality analysis were performed to identify essential proteins within a network. The higher degree in this network leads to higher interactions between proteins and causes a considerable effect. The most highly connected proteins in the PPI network are the most remarkable for their employment. The results of this study showed that three significant proteins including prostaglandin endoperoxide synthase 2 (PTGS2), TEK receptor tyrosine kinase (TEK), and fibroblast growth factor 18 (FGF18) were involved simultaneously in osteogenesis, angiogenesis, and their positive regulatory. Regarding the available literature, the results of this study confirmed that PTGS2 and FGF18 could be used as a GF in bone tissue engineering (BTE) applications to promote angiogenesis and osteogenesis. Nevertheless, TEK was not used in BTE applications until now and should be considered in future works to be examined in-vitro and in-vivo.


Assuntos
Osso e Ossos/fisiologia , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Neovascularização Fisiológica , Osteogênese , Biologia de Sistemas , Engenharia Tecidual , Regeneração Óssea/genética , Osso e Ossos/efeitos dos fármacos , Ontologia Genética , Fases de Leitura Aberta/genética , Mapas de Interação de Proteínas
10.
J Ophthalmic Vis Res ; 16(1): 56-67, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33520128

RESUMO

PURPOSE: Considering the significance of retinal disorders and the growing need to employ tissue engineering in this field, in-silico studies can be used to establish a cost-effective method. This in-silico study was performed to find the most effective growth factors contributing to retinal tissue engineering. METHODS: In this study, a regeneration gene database was used. All 21 protein-coding genes participating in retinal regeneration were considered as a protein-protein interaction (PPI) network via the "STRING App" in "Cytoscape 3.7.2" software. The resultant graph possessed 21 nodes as well as 37 edges. Gene ontology (GO) analysis, as well as the centrality analysis, revealed the most effective proteins in retinal regeneration. RESULTS: According to the biological processes and the role of each protein in different pathways, selecting the correct one is possible through the information that the network provides. Eye development, detection of the visible light, visual perception, photoreceptor cell differentiation, camera-type eye development, eye morphogenesis, and angiogenesis are the major biological processes in retinal regeneration. Based on the GO analysis, SHH, STAT3, FGFR1, OPN4, ITGAV, RAX, and RPE65 are effective in retinal regeneration via the biological processes. In addition, based on the centrality analysis, four proteins have the greatest influence on retinal regeneration: SHH, IGF1, STAT3, and ASCL1. CONCLUSION: With the intention of applying the most impressive growth factors in retinal engineering, it seems logical to pay attention to SHH, STAT3, and RPE65. Utilizing these proteins can lead to fabricate high efficiency engineered retina via all aforementioned biological processes.

11.
J Biomed Mater Res A ; 109(9): 1588-1599, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-33634587

RESUMO

The main aim of this study was to evaluate the efficacy of cerium oxide nanoparticles (CNPs) encapsulated in fabricated hybrid silk-fibroin (SF)/polycaprolactone (PCL) nanofibers as an artificial neural guidance conduit (NGC) applicable for peripheral nerve regeneration. The NGC was prepared by PCL and SF filled with CNPs. The mechanical properties, contact angle, and cell biocompatibility experiments showed that the optimized concentration of CNPs inside SF and SF/PCL wall of conduits was 1% (wt/wt). The SEM image analysis showed the nanoscale texture of the scaffold in different topologies depend on composition with fiber diameters at about 351 ± 54 nm and 420 ± 73 nm respectively for CNPs + SF and CNPs + SF/PCL fibrous mats. Furthermore, contact angle measurement confirmed the hydrophilic behavior of the membranes, ascribable to the SF content and surface modification through modified methanol treatment. The balance of morphological and biochemical properties of hybrid CNPs 1% (wt/wt) + SF/PCL construct improves cell adhesion and proliferation in comparison with lower concentrations of CNPs in nanofibrous scaffolds. The release of CNPs 1% (wt/wt) from both CNPs + SF and CNPs+ SF/PCL fibrous mats was highly controlled and very slow during the extended time of incubation until 60 days. Fabricated double-layered NGC using CNPs + SF and CNPs + SF/PCL fibers was consistent for application in nervous tissue engineering and regenerative medicine from a structural and biocompatible perspective.


Assuntos
Cério/farmacologia , Fibroínas/farmacologia , Nanopartículas/química , Tecido Nervoso/transplante , Poliésteres/farmacologia , Engenharia Tecidual , Animais , Bombyx , Proliferação de Células/efeitos dos fármacos , Preparações de Ação Retardada/farmacologia , Masculino , Nanofibras/química , Nanofibras/ultraestrutura , Nanopartículas/ultraestrutura , Ratos Wistar , Espectroscopia de Infravermelho com Transformada de Fourier , Resistência à Tração , Tecidos Suporte/química , Água
12.
Platelets ; 32(2): 183-188, 2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33577378

RESUMO

Bone tissue engineering (BTE) is a strategy for reconstructing bone lesions, which is rapidly developing in response to higher demands for bone repairing. Recently, this method, along with the emergence of functionally graded, biocompatible and biodegradable materials, has been expanded. Moreover, scaffolds with chemical, physical and external patterns have induced bone regeneration. However, the maintenance of healthy bone and its regeneration in the human body needs a series of complex and accurate processes. Hence, many studies have been accompanied for reconstructing bone by using blood-derived biomaterials, especially platelet-rich fabricates. The most important reason for using platelet-rich formulations in bone regeneration is based on releasing growth factors from alpha granules in platelets, which can induce osteogenesis. Moreover, the presence of fibrin nano-fiber structures as a constituent can provide a good substrate for cell attachments. This study attempts to review the history, structure, and biology of platelet-rich fibrin (PRF) as well as in vitro, pre-clinical, and clinical studies on the use of PRF for bone regeneration.


Assuntos
Regeneração Óssea/fisiologia , Fibrina Rica em Plaquetas/metabolismo , Engenharia Tecidual/métodos , Humanos
13.
J Biomed Mater Res B Appl Biomater ; 109(9): 1259-1270, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-33354913

RESUMO

The aim of this study was to prepare a porous scaffold out of 58S bioactive glass as the bare and coated with Zein to improve mechanical properties and acting as a carrier for Kaempferol controlled delivery. Porosity and morphology, mechanical properties, drug release behavior, bioactivity, cell attachment, and biodegradation of the scaffolds were evaluated accordingly. Obtained results indicated that the scaffolds coated by (7wt/v %) Zein solution, showed the highest mechanical strength (3.06 ± 0.4 MPa) and desirable porous morphology. These scaffolds could support bioactivity, cell attachment, and provide sustained drug release in the safe range of Kaempferol concentration confirmed via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis. Overall, this study showed that the Zein-coated scaffold possesses superior properties rather than bare scaffold, and the scaffolds coated with 7wt/v % Zein solution could be considered as appropriate scaffolds for bone regeneration.


Assuntos
Osso e Ossos/química , Cerâmica/química , Vidro/química , Quempferóis/química , Tecidos Suporte/química , Zeína/química , Adesão Celular , Linhagem Celular , Proliferação de Células , Materiais Revestidos Biocompatíveis/química , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Quempferóis/farmacologia , Fenômenos Mecânicos , Porosidade , Propriedades de Superfície
14.
Sci Rep ; 10(1): 14853, 2020 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-32908157

RESUMO

In this study, we aimed to fabricate osteoconductive electrospun carbon nanofibers (CNFs) decorated with hydroxyapatite (HA) crystal to be used as the bone tissue engineering scaffold in the animal model. CNFs were derived from electrospun polyacrylonitrile (PAN) nanofibers via heat treatment and the carbonized nanofibers were mineralized by a biomimetic approach. The growth of HA crystals was confirmed using XRD, FTIR, and EDAX analysis techniques. The mineralization process turned the hydrophobic CNFs (WCA: 133.5° ± 0.6°) to hydrophilic CNFs/HA nanocomposite (WCA 15.3° ± 1°). The in vitro assessments revealed that the fabricated 24M-CNFs nanocomposite was biocompatible. The osteoconductive characteristics of CNFs/HA nanocomposite promoted in vivo bone formation in the rat's femur defect site, significantly, observed by computed tomography (CT) scan images and histological evaluation. Moreover, the histomorphometric analysis showed the highest new bone formation (61.3 ± 4.2%) in the M-CNFs treated group, which was significantly higher than the negative control group (the defect without treatment) (< 0.05). To sum up, the results implied that the fabricated CNFs/HA nanocomposite could be considered as the promising bone healing material.


Assuntos
Carbono , Durapatita , Nanocompostos , Nanofibras , Engenharia Tecidual , Tecidos Suporte , Animais , Materiais Biomiméticos , Regeneração Óssea , Carbono/química , Carbono/farmacologia , Linhagem Celular , Durapatita/química , Durapatita/farmacologia , Humanos , Masculino , Nanocompostos/química , Nanocompostos/uso terapêutico , Nanofibras/química , Nanofibras/uso terapêutico , Osteogênese , Ratos , Ratos Wistar
15.
J Tissue Viability ; 29(4): 359-366, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32839065

RESUMO

The main goal of this study was to explore the beneficial effect of nerve growth factor (NGF)-overexpressing of human adipose-derived mesenchymal stem cells (hADSCs) encapsulated in injectable chitosan/ß-glycerophosphate/hydroxyethylcellulose (CS/ß-GP/HEC) hydrogel for spinal cord regeneration. The CS/ß-GP/HEC hydrogel and genetically transduced hADSCs using pseudo-lentiviruses-NGF were prepared. The mechanical properties, morphology and cytotoxicity of the hydrogel were investigated by rheometry, scanning electron microscope (SEM), and MTT assay, respectively. Rats animals were undergone spinal cord injury (SCI), then one-week post-injury, CS/ß-GP/HEC hydrogel, transduced hADSCs and transduced hADSCs/CS/ß-GP/HEC hydrogel injected into the site of the lesion. Animals with SCI and animals with laminectomy without SCI were considered as negative control and sham groups, respectively. Positive control group received no surgical intervention. At eight weeks post-injection, histological studies indicated a significant increase in cell proliferation, a smaller cavity in size at the SCI site as well as better locomotor functions for transduced hADSCs/CS/ß-GP/HEC hydrogel group (P ≤ 0.05) compared to other experimental groups. Our results showed that CS/ß-GP/HEC hydrogel in combination with transduced-hADSCs is able to successfully regenerate SCI. These results may be applicable in the selection of the best therapeutic strategy based on gene therapy and tissue engineering for SCI treatment.


Assuntos
Hidrogéis/administração & dosagem , Fator de Crescimento Neural/farmacologia , Regeneração da Medula Espinal/efeitos dos fármacos , Animais , Quitosana/administração & dosagem , Quitosana/farmacologia , Quitosana/uso terapêutico , Modelos Animais de Doenças , Hidrogéis/farmacologia , Hidrogéis/uso terapêutico , Injeções/métodos , Fator de Crescimento Neural/uso terapêutico , Ratos , Espectrofotometria Infravermelho/métodos
16.
ACS Biomater Sci Eng ; 6(11): 6285-6298, 2020 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-33449643

RESUMO

The human amniotic membrane (HAM) has been viewed as a potential regenerative material for a wide variety of injured tissues because of its collagen-rich content. High degradability of HAM limits its wide practical application in bone tissue engineering. In this study, the natural matrix of the decellularized amniotic membrane was developed by the double diffusion method. The results confirmed a reduction of the amniotic membrane's degradability because of the deposition of calcium and phosphate ions during the double diffusion process. Real-time PCR results showed a high expression of osteogenesis-related genes from adipose-derived mesenchymal stem cells (ADMSCs) cultured on the surface of the developed mineralized amniotic membrane (MAM). Further in vivo experiments were conducted using an MAM preseeded with ADMSCs and a critical-size rat calvarial defect model. Histopathological results confirmed that the MAM + cell sample has excellent potential in bone regeneration.


Assuntos
Âmnio , Engenharia Tecidual , Animais , Biomimética , Regeneração Óssea , Diferenciação Celular , Humanos , Ratos
17.
Biofabrication ; 12(1): 015021, 2019 12 19.
Artigo em Inglês | MEDLINE | ID: mdl-31658444

RESUMO

Mesenchymal stem cells (MSCs) are considered primary candidates for treating complex bone defects in cell-based therapy and tissue engineering. Compared with monolayer cultures, spheroid cultures of MSCs (mesenspheres) are favorable due to their increased potential for differentiation, extracellular matrix (ECM) synthesis, paracrine activity, and in vivo engraftment. Here, we present a strategy for the incorporation of microparticles for the fabrication of osteogenic micro-tissues from mesenspheres in a cost-effective and scalable manner. A facile method was developed to synthesize mineral microparticles with cell-sized spherical shape, biphasic calcium phosphate composition (hydroxyapatite and ß-tricalcium phosphate), and a microporous structure. Calcium phosphate microparticles (CMPs) were incorporated within the mesenspheres through mixing with the single cells during cell aggregation. Interestingly, the osteogenic genes were upregulated significantly (collagen type 1 (Col 1) 30-fold, osteopontin (OPN) 10-fold, and osteocalcin (OCN) 3-fold) after 14 days of culture with the incorporated CMPs, while no significant upregulation was observed with the incorporation of gelatin microparticles. The porous structure of the CMPs was exploited for loading and sustained release of an angiogenic small molecule. Dimethyloxaloylglycine (DMOG) was loaded efficiently onto the CMPs (loading efficiency: 65.32 ± 6%) and showed a sustained release profile over 12 days. Upon incorporation of the DMOG-loaded CMPs (DCMPs) within the mesenspheres, a similar osteogenic differentiation and an upregulation in angiogenic genes (VEGF 5-fold and kinase insert domain (KDR) 2-fold) were observed after 14 days of culture. These trends were also observed in immunostaining analysis. To evaluate scalable production of the osteogenic micro-tissues, the incorporation of microparticles was performed during cell aggregation in a spinner flask. The DCMPs were efficiently incorporated and directed the mesenspheres toward osteogenesis and angiogenesis. Finally, the DCMP mesenspheres were loaded within a three-dimensional printed cell trapper and transplanted into a critical-sized defect in a rat model. Computed tomography and histological analysis showed significant bone formation with blood vessel reconstruction after 8 weeks in this group. Taken together, we provide a scalable and cost-effective approach for fabrication of osteogenic micro-tissues, as building blocks of macro-tissues, that can address the large amounts of cells required for cell-based therapies.


Assuntos
Células-Tronco Mesenquimais/citologia , Engenharia Tecidual/métodos , Animais , Bioimpressão/economia , Proliferação de Células , Matriz Extracelular/química , Matriz Extracelular/metabolismo , Humanos , Células-Tronco Mesenquimais/química , Células-Tronco Mesenquimais/metabolismo , Osteocalcina/metabolismo , Osteogênese , Ratos , Ratos Wistar , Engenharia Tecidual/economia , Engenharia Tecidual/instrumentação , Tecidos Suporte/química , Tecidos Suporte/economia
18.
Mater Sci Eng C Mater Biol Appl ; 105: 110071, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31546377

RESUMO

In the current study, in vitro analysis of the osteogenic potential of different scaffolds based on strontium-substituted ß-TCP (Sr-TCP) and bioactive glass (BG) ceramics was conducted using rabbit bone marrow-derived mesenchymal stem cells (rBMSCs) and the osteogenic ability of the prepared Sr-TCP and BG scaffold was evaluated through alkaline phosphatase activity, mineral deposition by Alizarin red staining, and osteoblastic gene expression experiments. The obtained in vitro results revealed that among experimental Sr-TCP/BG nanocomposite scaffold samples with the composition of Sr-TCP/BG: 100/0, 50/50, 75/25, and 25/75, the 50Sr-TCP/50BG sample presented better osteoinductive properties. Therefore, the optimized 50Sr-TCP/50BG nanocomposite scaffold was chosen for further in vivo experiments. In vivo implantation of 50Sr-TCP/50BG scaffold and hydroxyapatite (HA)/TCP granules in a rabbit calvarial defect model showed slow degradation of 50Sr-TCP/50BG scaffold and high resorption rate of HA/TCP granules at 5 months' post-surgery. However, the 50Sr-TCP/50BG scaffolds loaded by mesenchymal stem cells (MSCs) were mainly replaced with new bone even at 2 months post-implantation. Based on the obtained engineering and biological results, 50Sr-TCP/50BG nanocomposite scaffold containing MSCs could be considered as a promising alternative substitute even for load-bearing bone tissue engineering applications.


Assuntos
Células da Medula Óssea/metabolismo , Fosfatos de Cálcio/química , Células Imobilizadas , Cerâmica/química , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Nanocompostos/química , Estrôncio/química , Tecidos Suporte/química , Animais , Células da Medula Óssea/patologia , Células Imobilizadas/metabolismo , Células Imobilizadas/patologia , Células Imobilizadas/transplante , Células-Tronco Mesenquimais/patologia , Porosidade , Coelhos
19.
Prog Biomater ; 8(1): 31-42, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30706299

RESUMO

A hybrid hydrogel was obtained from decellularized extract from Wharton's jelly (DEWJ) and silk fibroin (SF) and characterized for cartilage tissue engineering. Wharton's jelly was used due to its similarity with articular cartilage in extracellular matrix composition. Also, silk fibroin has good mechanical properties which make this construct appropriate for cartilage repair. Decellularization of Wharton's jelly was verified by DAPI staining, DNA quantification, and PCR analysis. Then, the biochemical composition of DEWJ was determined by ELISA kits for total proteins, collagens, sulfated glycosaminoglycans (sGAG), and transforming growth factor ß1 (TGF-ß1). After fabricating pure SF and SF/DEWJ hybrid hydrogels, their physical and mechanical properties were characterized by FESEM, Fourier-transform infrared spectroscopy (FTIR) and rheological assays (amplitude and frequency sweeps). Furthermore, cell viability and proliferation were assessed by MTT assay. The results have shown that DEWJ in hybrid hydrogels enhances mechanical properties of the construct relative to pure SF hydrogels. Also, this extract at its 40% concentration in culture media and 20% or 40% concentrations in SF/DEWJ hybrid hydrogels significantly increases population of the cells compared to control and pure SF hydrogel after 7 days. In conclusion, this study proposes the potential of SF/DEWJ hybrid hydrogels for cartilage tissue engineering applications.

20.
Cell Biol Int ; 2019 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-30811084

RESUMO

The mechanical property of bone tissue scaffolds is one of the most important aspects in bone tissue engineering that has remained problematic. In our previous study, we fabricated a three-dimensional scaffold from nano-hydroxyapatite/gelatin (nHA/Gel) and investigated its efficiency in promoting bone regeneration both in vitro and in vivo. In the present study, the effect of adding silicon carbide (SiC) on the mechanical and biological behaviors of the nHA/Gel/SiC and bone regeneration in vivo were determined. nHA and SiC were synthesized and characterized by the X-ray diffraction pattern and transmission electron microscope image. Layer solvent casting, freeze drying, and lamination techniques were applied to prepare these scaffolds. Then, the biocompatibility and cell adhesion behavior of the synthesized nHA/Gel/SiC scaffolds were investigated. For in vivo studies, rats were categorized into three groups: blank defect, blank scaffold, and rat bone marrow mesenchymal stem cells (rBM-MSCs)/scaffold. After 1, 4, and 12 weeks post-injury, the rats were sacrificed and the calvaria were harvested. Sections with a thickness of 5 µm thickness were prepared and stained with hematoxylin-eosin and Masson's Trichrome, and immunohistochemistry was performed. Our results showed that SiC effectively increased the mechanical properties of the nHA/Gel/SiC scaffold. No significant differences were observed in biocompatibility, cell adhesion, and cytotoxicity of the nHA/Gel/SiC in comparison with the nHA/Gel nanocomposite. Based on histological and immunohistochemical studies, both osteogenesis and collagenization were significantly higher in the rBM-MSCs/scaffold group, quantitatively and qualitatively. The present study strongly suggests the potential of SiC as an alternative strategy to improve the mechanical and biological properties of bone tissue engineering scaffolds, and shows that the pre-seeded nHA/Gel/SiC scaffold with rBM-MSCs improves osteogenesis in the engineered bone implant.

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