RESUMO
The undesirable inflammation and the excessive M1 macrophage activity may lead to inflammatory diseases. Corticosteroids and stem cell therapy are used in clinical practice to promote anti-inflammatory responses. However, this protocol has limitations and is associated with numerous side effects. In this study, the synergistic anti-inflammatory effects of dexamethasone (Dex) and mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) were evaluated to enhance the polarization of M1 inflammatory macrophages into the anti-inflammatory (M2) phenotype. Hence, we designed different combinations of Dex and EVs using three methods, including EVs isolated from Dex-preconditioned MSCs (Pre-Dex-EVs), EVs loaded with Dex (L-Dex-EVs), and EVs and Dex co-administration (Dex + EVs). All designed EVs had a significant effect on reducing the expression of M1-related genes (iNOS, Stat1, and IRF5), cytokines (IL6 and TNF-a), and CD markers (CD86) in lipopolysaccharide-stimulated macrophages. On the other hand, these combinations promoted the expression of alternative-activated M2-related genes (Arg-1, Stat6, and IRF4), cytokine (IL10), and CD markers (CD206).The combination of Dex and MSC-EVs enhances the effectiveness of both and synergistically promotes the conversion of inflammatory macrophages into an anti-inflammatory state.
Assuntos
Vesículas Extracelulares , Células-Tronco Mesenquimais , Citocinas/metabolismo , Células-Tronco Mesenquimais/metabolismo , Vesículas Extracelulares/metabolismo , Macrófagos , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/metabolismo , Dexametasona/farmacologiaRESUMO
Among medicinal plants, Acridocarpus orientalis (AO) possesses a remarkable anti-cancer potential, possibly because of its anti-oxidant property. In this study, the leaf and stem extracts from AO were assessed to find the bioactive compound with selective anti-cancer properties. The MTT viability and live and dead assays revealed that around 80% and 98% of 4T1 cells survival were declined after 48 h incubation with leaf and stem extracts, respectively. The leaf extract increased stem cell proliferation by 20% whereas the stem extract inhibited around 22% of stem cells proliferation after 48 h treatment. The live and dead assay of MSCs confirmed that 40% of the MSCs died when treated with AO stem extract. On the other hand, there were no dead cells after two days of treatment with the leaf extract. Followed by the induction of cell cycle arrest in G0/G1-phase, the real-time PCR demonstrated apoptosis properties in 4T1 cells through overexpression of Bax and down-regulation of BCL2 genes. Interestingly, within the pure compounds isolated from AO leaf extract, Morin was responsible for the inhibition of 4T1 cells proliferation as well as MSCs expansion, predicting to play an essential role in the treatment of cancer. The promising in vitro anti-cancer and stem cell-inductive properties of morin isolated from AO extract may provide a great potential to produce selective herbal derived drugs.
Assuntos
Malpighiaceae/metabolismo , Extratos Vegetais/farmacologia , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Folhas de Planta/metabolismo , Caules de Planta/metabolismo , Plantas Medicinais/metabolismoRESUMO
Coculture systems are widely used in tissue engineering to mimic cell-cell interactions between different populations. This study aimed to find an improved and convenient system for the corneal epithelial differentiation of conjunctiva derived mesenchymal stem cells (CJMSCs). Thus, the cells were used to reconstruct corneal epithelial cells. Obtained by flow cytometry data, 51.9% of isolated CJMSCs were immune reactive for SSEA4+ antibody which are more potent to differentiate into corneal epithelial cells. A differential medium in a single culture plate was applied and compared to a Coculture with a SHEM medium and a Coculture with a commercial medium. It was found that CJMSCs can be induced to corneal epithelial cells through in vitro co-culturing in a SHEM medium; this was confirmed with immunostaining results. Moreover, relative gene expression results showed that a Coculture system with a SHEM medium can provide a more favorable microenvironment for cells to differentiate into epithelial cells than a single culture or a Coculture with a CnT-Prime commercial medium. Finally, as platforms for cell differentiation, CJMSCs can differentiate into epithelial lineages. This was proven using immunofluorescence staining.
Assuntos
Diferenciação Celular , Túnica Conjuntiva/metabolismo , Células Epiteliais/metabolismo , Epitélio Corneano/metabolismo , Células-Tronco Mesenquimais/metabolismo , Engenharia Tecidual , Técnicas de Cocultura , Túnica Conjuntiva/citologia , Células Epiteliais/citologia , Epitélio Corneano/citologia , Humanos , Células-Tronco Mesenquimais/citologia , Nicho de Células-TroncoRESUMO
Tissue engineering is a promising emerged method trying to reconstruct lost tissues that using synthetic and biomaterials and their combination with cells. The purpose of this study is increase osteoinductivity of polyethersulfone (PES) by using polyaniline (PANi). In this study, after fabrication of PES and composited PES-PANi scaffolds by electrospinning, scaffolds were characterized morphologically and mechanically. Then osteoinductivity of scaffolds was investigated by osteogenic differentiation of human mesenchymal stem cells (MSCs) cultured on the PES and PES-PANI in comparison to the tissue culture polystyrene as a control. The Osteogenic potential of MSCs was evaluated by Alizarin Red staining, ALP activity, calcium content assay and bone related gene expression assay. Scaffolds were smooth, bead free and in the scale of nanometers and PES mechanical stability was decreased significantly after composite with PANi. Highest growth, ALP activity and deposited calcium of cells were observed in the PES-PANi groups compared to the PES and TCPS. Significantly higher expression of collagen type one and osteonectin was also detected in the PES-PANi group. It can be concluded that PES-PANi construct has potential to be a good candidate as bone grafting substitute and using in tissue engineering applications.
Assuntos
Compostos de Anilina/química , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Proliferação de Células , Células-Tronco Mesenquimais/metabolismo , Nanofibras/química , Osteogênese , Polímeros/química , Sulfonas/química , Humanos , Células-Tronco Mesenquimais/citologiaRESUMO
The miR-17-92 cluster consisted of seven miRNAs (mir-17-5p, -17-3p, -18a, -19a, -20a, -19b-1, and -92a-1). Previous studies have shown this cluster has been over-expressed in several cancers. The aim of this study was to evaluate the over-expression impacts of miR-17-92 on stem cells. In the current work, the effect of miR-17-92 cluster which was cloned in Lentiviral vector has been investigated on unrestricted somatic stem cells (USSCs). Tumor suppressor genes (p53, p15, RBL1, SMAD2, SMAD4, and MAPK-1) expression, especially p53, was considerably reduced. These data show the potential of miR-17-92 for oncogenesis regulation in stem cells. In conclusion, the role of miR-17-92 in USSCs may provide a better understanding of its function in tumorigenesis and for the possible use in cell therapy of the anti-mir-17-92 cluster.
Assuntos
Regulação da Expressão Gênica , Genes Supressores de Tumor , Células-Tronco Hematopoéticas/metabolismo , MicroRNAs/genética , Família Multigênica , Ciclo Celular/genética , Células Cultivadas , Inibidor de Quinase Dependente de Ciclina p15/genética , Sangue Fetal/citologia , Humanos , Proteína Quinase 1 Ativada por Mitógeno/genética , RNA Longo não Codificante , Proteína p107 Retinoblastoma-Like/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína Smad2/genética , Proteína Smad4/genética , Proteína Supressora de Tumor p53/genéticaRESUMO
Postoperative adhesion can cause complications, such as pain and organ blockage, in the abdominal regions. To address this issue, surgical techniques and antiadhesive treatments are applied. Given the significant role of vascularization in adhesion band formation, Avastin (Ava) that targets vascular endothelial growth factor (VEGF) can be applied to prevent peritoneal adhesion bands. Moreover, Alginate (Alg), a natural polysaccharide, is a promising physical barrier to prevent adhesion bands. Incorporating Ava into Alg hydrogel in a form of 3D-printed scaffold (Alg/Ava) has potential to suppress inflammation and angiogenesis, leading to reduce peritoneal adhesion bands. Following physical, morphological, and biocompatibility evaluations, the efficacy of Alg and Ava alone and their combination in Alg/Ava on the formation of postsurgical adhesions is evaluated. Upon confirming physical stability and sustained release of Ava, the Alg/Ava scaffold effectively diminishes both the extent and strength of adhesion bands. Histopathological examination shows that the reduction in fibrosis and inflammation is responsible for preventing adhesion bands by the Alg/Ava scaffold. Additionally, the cytokine assessment reveals that this is due to the inhibition in the secretion of VEGF and Interleukin 6 suppressing vascularization and inflammatory pathways. This study suggests that a 3D-printed Alg/Ava scaffold has great potential to prevent the postsurgical adhesion bands.
Assuntos
Alginatos , Bevacizumab , Impressão Tridimensional , Alicerces Teciduais , Fator A de Crescimento do Endotélio Vascular , Alginatos/química , Alginatos/farmacologia , Aderências Teciduais/prevenção & controle , Animais , Alicerces Teciduais/química , Bevacizumab/farmacologia , Bevacizumab/química , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ratos , Complicações Pós-Operatórias/prevenção & controle , Humanos , Hidrogéis/química , Hidrogéis/farmacologiaRESUMO
ABSTRACT: Background: Sepsis is a life-threatening disorder that leads to the induction of inflammatory responses and organ failure. Phage therapy is a new approach to controlling infections resistant to common treatments, including sepsis. Several studies have shown the effect of lytic bacteriophages on infection control by reducing the bacterial load. The present study deals with lysogenic bacteriophage M13 on the inflammatory responses caused by cecal ligation and puncture (CLP)-induced sepsis in a mouse model. Methods Bacteriophage M13 harvested from ER2738, titrated, and confirmed by transmission electron microscopy analysis. In vitro toxicity and immunomodulatory effect of bacteriophage M13 were assessed on splenocytes by measurement of cell viability and the production level of cytokines, nitric oxide, and reactive oxygen species. For in vivo experiments, 8-weeks-old male C57BL/6 mice were randomly divided into the following three groups: CLP + NS (treated with normal saline), CLP + M13 (treated with an intraperitoneal injection of 10 9 PFU/mL of bacteriophage M13), and sham + NS (induced surgery but without ligation and puncture, treated with NS). The mice were killed at different time points after surgery (6, 24, 48, and 72, n = 10 for each time point of each group). The kidney, liver, and lungs were harvested for histopathological analysis, and blood was obtained for cytokine and liver enzyme assay. The spleen was used to assess the bacterial load using colony-forming unit assay. The rectal temperature and survival were evaluated during the study. Results According to the in vitro results, 10 9 PFU/mL of bacteriophage M13 was not toxic and did not affect the level of cytokine, nitric oxide, and reactive oxygen species production by splenocytes, but it reduced the inflammatory response of splenocytes in responses to LPS. In vivo studies indicated that the amount of proinflammatory cytokines, liver enzymes, bacterial load, and organ failure were decreased in the CLP + M13 group compared with CLP + NS, whereas the survival rate was increased. Conclusions These experiments demonstrated that bacteriophage M13 could lessen the consequences related to sepsis in CLP mice and can be considered a therapeutic approach in sepsis.
Assuntos
Bacteriófago M13 , Sepse , Camundongos , Masculino , Animais , Óxido Nítrico , Espécies Reativas de Oxigênio , Camundongos Endogâmicos C57BL , Citocinas , Sepse/tratamento farmacológico , Punções/efeitos adversos , Ceco/cirurgia , Modelos Animais de DoençasRESUMO
This study aimed to develop a local 3 D-printed bioactive graft using poly-caprolacton (PCL) as a drug carrier and 3-O-acetyl-ß-boswellic acid (ß-ABA) as an anticancer compound. ß-ABA-loaded 3 D-printed scaffold was fabricated and physically characterized. The results indicated more desirable mechanical and physical properties of the ß-ABA-loaded PCL mat in comparison with the PCL scaffold. Following sustained release of ß-ABA, the ß-ABA-loaded PCL scaffold revealed selective cytotoxic activity against melanoma cells, while the PCL + ABA with the bolus delivery of ß-ABA was toxic against fibroblast cells. Followed by the induction of apoptosis in melanoma cells at the gene level, the result of the western blot showed that the ß-ABA-loaded scaffold significantly up-regulated P53 and down-regulated BCL2, with an increment in the ratio of Bax/BCL2. The selective anti-cancer properties of ß-ABA-loaded 3 D printed scaffold against melanoma cells indicated that this scaffold could be potentially used as a bioactive graft to improve the melanoma treatment.
Assuntos
Melanoma , Humanos , Melanoma/tratamento farmacológico , Impressão Tridimensional , Proteínas Proto-Oncogênicas c-bcl-2RESUMO
A tridentate ligand LH3 ((2-hydroxy-3-methoxybenzylidene)-2-(hydroxyimino)propanehydrazide) comprising o-vanillin, hydrazone and oxime donor groups has been employed to prepare a series of tetranuclear Ln(III) complexes. The reaction of ligand LH3 with Ln(NO3)3 [Ln = Sm, Eu, Gd, Tb, Dy, Ho, Er] in MeOH yielded Ln4(LH)6(MeOH)2 (Ln = Sm(1), Eu(2), Gd(3), Tb(4), Ho (6) and Er (7))] whereas the corresponding reaction with Dy(NO3)3 afforded Dy4(LH)4(LH2)2(OH)2 (5). All complexes were characterized by various analytical techniques including single crystal X-ray diffraction, IR spectroscopy, UV-Vis spectroscopy, and elemental analysis. To investigate the potential of these lanthanide complexes for wound healing applications, their effects on fibroblast viability, migration, and M2 macrophage polarization were evaluated. The cytotoxicity assessment revealed that complexes 2(Eu), 4(Tb), 5(Dy), and 7(Er) significantly enhanced fibroblast viability compared to the negative control (NC). In vitro wound healing assay demonstrated that complexes 2(Eu) and 7(Eu) substantially promoted fibroblast migration compared to the NC. Moreover, complex 2(Eu) exhibited significant anti-inflammatory effects by reducing the phagocytic ability of lipopolysaccharide (LPS)-stimulated macrophage cells and attenuating nitric oxide (NO) production. In conclusion, among the series of complexes tested, complex 2(Eu) displayed the most potent anti-inflammatory effect on macrophage cells, while simultaneously promoting fibroblast viability and migration. This unique combination of properties renders complex 2 (Eu) highly promising for wound healing applications.
Assuntos
Elementos da Série dos Lantanídeos , Elementos da Série dos Lantanídeos/farmacologia , Elementos da Série dos Lantanídeos/química , Ligantes , Fibroblastos , Macrófagos , Anti-InflamatóriosRESUMO
This study aimed to compare the effect of Boswellic acid derivatives on the viability, apoptosis, and epigenomic profiling of breast cancer. According to the viability assays, 3-O-acetyl-11-keto-ß-Boswellic acid (AKBA) showed more toxicity against MDA-MB-231 cells when compared with the 3-O-acetyl-ß-Boswellic acid (ABA). In contrast, ABA revealed less toxicity against MCF-10A. Cell cycle and apoptosis assays determined the maximum apoptotic effect of AKBA on MCF-7, and MDA-MB-231 cells. Interestingly, ß-Boswellic acid (BA) and ABA did not promote the apoptosis in MCF-10A cells. Transwell migration assay indicated the greatest normalized inhibition (around 160%) in the migration of MDA-MB-231 cells induced by AKBA. The expression of P53, BAX, and BCL2 genes in cancerous cell lines has affirmed that both AKBA and ABA could induce the maximal apoptosis. Western-blot investigation demonstrated that the maximum over-expression of P53 protein (1.96 times) was caused by AKBA in MDA-MB-231 cells, followed by ABA in MCF-7 cells. The BCL2 protein expression was in agreement with the previously reported results. The global DNA methylation in both cancerous cells was reduced by ABA. These results suggest that ABA represented more epigenetic modulatory effect while AKBA shows more cytotoxic and apoptotic effect against breast cancer cell lines.
Assuntos
Epigenômica , Neoplasias , Proteína Supressora de Tumor p53 , Proteínas Proto-Oncogênicas c-bcl-2 , Epigênese GenéticaRESUMO
Background: and purpose: The formation of postoperative intra-abdominal adhesion band formation may lead to severe complications. This study aimed to evaluate the preventive effect of local administration of frankincense n-hexane extract (FHE) on the formation of postsurgical adhesion bands. Materials and methods: FHE was extracted from the resin of a Boswellia sacra tree and its components were identified by gas chromatography-mass spectrometry (GC-MS). In an animal model, the expression levels of TNF-α and TGF-ß1 cytokines after application of FHE were assessed to check the inflammatory and fibrotic cues, respectively. Results: Following FHE compound analysis, in vivo experiments demonstrated that intraoperative local administration of FHE resulted in the prevention of adhesion band formation. The adhesion grades in the FHE-treated group were significantly lower than those in the negative control (NC) and the positive control (Interceed). The infiltration of inflammatory cells observed by histopathology revealed a significant anti-inflammatory potential of FHE. Furthermore, the gene expression results proved that significant suppression of TNF-α and TGF-ß1 was responsible for its antiadhesion properties. Conclusions: The study reported the potential of FHE as an ointment for the prevention of adhesion bands.
RESUMO
BACKGROUND: This study was designed to evaluate the effect of different concentrations of conjugated equine estrogens (CEE) on the ovarian epithelium of female CD1 mice. METHODS: Twenty-four female mice at 7 months with irregular estrus cycles were randomly divided into four groups of 6 mice each. Group one was considered as a control group and received a daily dose of 0.5ml of propylene glycol, for three weeks, while those in the treatment groups received a daily dose of 14µg/kg, 28µg/kg and 56µg/kg conjugated equine estrogens, respectively. RESULTS: The results from this study showed a strong correlation between elevated concentrations of CEE and histological changes in ovarian surface epithelium (OSE). They also showed that administration of high-dose estrogen created the conditions for excessive proliferation of OSE which may progress into the development of cysts in the ovaries. CONCLUSION: This study concluded that high concentrations of CEE may increase the chances of developing epithelial ovarian cancer.
Assuntos
Estrogênios Conjugados (USP) , Ovário , Animais , Modelos Animais de Doenças , Epitélio , Estrogênios/farmacologia , Estrogênios Conjugados (USP)/farmacologia , Estrogênios Conjugados (USP)/uso terapêutico , Feminino , CamundongosRESUMO
The aim of this study was to evaluate a bioactive multilayer wound dressing, based on chitosan and alginate. To enhance healing potential, Dracaena Cinnabari and Aloe Vera were loaded as separate layers into the scaffold. The bare and bioactive multilayered scaffolds were fabricated by an iterative layering freeze-drying technique. Following of topographical, chemical, and physical assessment, the performance of the scaffolds was evaluated in vitro and in vivo. The results revealed adequate attachment, and proliferation of human foreskin fibroblasts, indicating excellent biocompatibility of the bioactive scaffold. In vivo, the performance of the multi-layered scaffold loaded with the bioactive materials was comparable with Comfeel plus®. The wounds treated with the bioactive scaffold exhibited superior hypergranulation, fibroblast maturation, epithelization, and collagen deposition, with minimal inflammation, and crust formation. It is concluded that the synergism of extracellular matrix-mimicking multi-layered scaffolding with Aloe Vera and Dracaena Cinnabari could be considered as a supportive wound dressing.
Assuntos
Alginatos/química , Bandagens , Quitosana/química , Extratos Vegetais/farmacologia , Alicerces Teciduais/química , Cicatrização/efeitos dos fármacos , Alginatos/farmacologia , Aloe/química , Animais , Proliferação de Células/efeitos dos fármacos , Quitosana/farmacologia , Colágeno/metabolismo , Dracaena/química , Matriz Extracelular/metabolismo , Fibroblastos/efeitos dos fármacos , Prepúcio do Pênis/patologia , Humanos , Masculino , Microscopia Eletrônica de Varredura/métodos , Extratos Vegetais/química , Ratos , Ratos WistarRESUMO
BACKGROUND: The development of anti-cancer drugs with the ability to inhibit brain metastasis through the blood-brain barrier (BBB) is substantially limited due to the lack of reliable in vitro models. MAIN METHODS: In this study, the Geltrex-based Transwell and microfluidic BBB models were applied to screen the effect of ß-boswellic acid (ß-BA) on the metastasis of MDA-MB-231 cells through the BBB in static and dynamic conditions, respectively. MAJOR RESULTS: The toxicity assay revealed that ß-BA deteriorates MDA-MB-231 cells, while ß-BA had no detectable toxic effects on human umbilical vein endothelial cells (HUVECs) and astrocytes. Trans-endothelial electrical resistance evaluation showed sustainable barrier integrity upon treatment with ß-BA. Vimentin expression in HUVECs, evaluated using western blot, confirmed superior barrier integrity in the presence of ß-BA. The obtained results were confirmed using an invasion study with a cell tracker and a scanning electron microscope. ß-BA significantly inhibited metastasis by 85%, while cisplatin (Cis), a positive control, inhibited cancer cell migration by 12% under static conditions. Upon applying a dynamic BBB model, it was revealed that ß-BA-mediated metastasis inhibition was significantly higher than that mediated by Cis. CONCLUSIONS AND IMPLICATIONS: In summary, the current study proved the anti-metastatic potential of ß-BA in both static and dynamic BBB models.
Assuntos
Barreira Hematoencefálica , Triterpenos , Células Endoteliais da Veia Umbilical Humana , Humanos , Microfluídica , Triterpenos/farmacologiaRESUMO
Stem cells can be obtained from a variety of sources. To compare the effect of cell source on the osteogenic differentiation potential, buccal fat pad-derived mesenchymal stem cells (BFP-MSCs), bone marrow-derived MSCs (BM-MSCs) and unrestricted somatic stem cells (USSCs) with different accessibility in time and region, were cultured on bioceramic (Bio-Oss®) coated electrospun polycaprolactone (PCL) scaffold (PCL-Bio). After scaffold characterization, stem cells proliferation and osteogenic differentiation were investigated by MTT and Alizarin red staining, alkaline phosphatase activity, calcium content and gene expression assays. Proliferation rate of the stem cells was not significantly different with each other, only USSCs showed significantly lower proliferation rate while cultured on PCL-Bio; although, PCL-Bio showed better proliferation support in comparison with tissue culture plate and PCL. Mineralization of the BM-MSCs was significantly higher than others, while BFP-MSCs were close to it. Highest ALP activity was detected in BFP-MSCs cultured on PCL-Bio. USSCs demonstrated higher gene expression level in three genes, although differences were not huge compared to others. According to the results and due to the availability, facilitated preparation procedure and less patients suffering, BFP-MSCs have a better choice than BM-MSCs and USSCs for use in bone tissue engineering.
Assuntos
Materiais Biocompatíveis/farmacologia , Diferenciação Celular/efeitos dos fármacos , Cerâmica/química , Osteogênese/efeitos dos fármacos , Poliésteres/química , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Materiais Biocompatíveis/química , Biomarcadores/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Nanofibras/química , Células-Tronco/metabolismoRESUMO
In current study we aimed to coat the PLLA scaffold with zinc (Zn) silicate mineral nanoparticles. Then, using equine adipose-derived stem cells (ASCs) we intended to compare the osteogenic induction potency of Zn silicate mineral-coated PLLA scaffold with uncoated PLLA scaffold and tissue culture plastic (TCPS). Adipose tissues were collected from 3 horses, and isolation of ASCs was achieved by enzymatic digestion. PLLA scaffold was successfully prepared using a phase separation method and coated with Zn silicate mineral nanoparticles. The coating efficiency was then characterized by scanning electron microscopy and further evaluated with the application of fourier transform infrared microscopic imaging. Viability and growth characteristics of ASCs on TCPS, uncoated and coated PLAA scaffolds were investigated by MTT assay. Alizarin Red staining was performed for determination of calcium deposition following the osteogenic induction. Furthermore, other common osteogenic markers such as alkaline phosphatase (ALP) activity, calcium content, as well as osteogenic (Runx2, ALP, osteonectin, and collagen I) marker genes were also evaluated. Our data showed that Zn silicate mineral nanoparticles was coated successfully on PLLA scaffold and such scaffold had no detrimental effect on cell growth rate as indicated by MTT assay. Moreover, ASCs that differentiated on Zn silicate mineral-coated PLLA scaffold indicated higher ALP activity, more calcium content, and higher expression of bone-related genes than that on uncoated PLLA scaffold and TCPS. Adequate proliferation rate and higher expression of osteogenic markers of stem cells, provides this scaffold as a suitable substrate to support proliferation and differentiation of ASCs in equine.
Assuntos
Cavalos/crescimento & desenvolvimento , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanopartículas Metálicas/administração & dosagem , Osteogênese/efeitos dos fármacos , Silicatos/administração & dosagem , Alicerces Teciduais/veterinária , Compostos de Zinco/administração & dosagem , Tecido Adiposo/citologia , Animais , Minerais/administração & dosagemRESUMO
Since ancient times, some herbal medicines have been extensively used for burn and wound treatments, showing preference to the common synthetic medications by virtue of having less side effects and faster healing rate. In this study, hybrid nanofibrous scaffolds of poly-l-lactic-acid (PLLA) and gelatin incorporated L. inermis were fabricated via electrospinning technique. Morphology and characteristics of the scaffolds were studied by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR), respectively. The release profile of the L. inermis from the nanofibers was also assessed in vitro. Moreover, the structural stability of the released L. inermis from the nanofibers was evaluated using high-performance liquid chromatography (HPLC). The nanofibers showed a gradual release of L. inermis up to two days while the intact structure was preserved. Furthermore, antibacterial assay demonstrated that L. inermis-loaded nanofibrous scaffolds could effectively kill E. coli and S. aureus within 2â¯h. Finally, biocompatibility of the nanofibers was proven on 3T3 fibroblasts. Therefore, the L. inermis loaded PLLA-Gelatin nanofibers showed a potential application as a wound dressing in order to control wound infections.
Assuntos
Anti-Infecciosos/farmacologia , Bandagens , Lawsonia (Planta) , Nanofibras , Infecção dos Ferimentos/prevenção & controle , Células 3T3 , Animais , Camundongos , Nanofibras/química , Poliésteres , Cicatrização/efeitos dos fármacosRESUMO
Migration of fibroblasts into wound area is a critical phenomenon in wound healing process. We used an appropriate system to fabricate an electrospun bioactive scaffold with controlled release of PDGF-BB in order to induce migration of primary skin fibroblast cells. First of all, protein-loaded chitosan nanoparticles based on ionic gelation interaction between chitosan and sodium tripolyphosphate were prepared. Then polycaprolactone electrospun fibers containing chitosan nanoparticles or PDGF-BB-loaded chitosan nanoparticles were prepared. Cellular attachment and morphology of cells seeded on scaffolds with or without PDGF-BB were evaluated by using a fluorescence microscope and scanning electron microscopy. Cells were well-oriented 72 h after seeding on the scaffolds containing PDGF-BB. The mean aspect ratio of populations on scaffold containing PDGF-BB-loaded chitosan nanoparticles was significantly greater than those on the scaffold containing chitosan nanoparticles but no PDGF-BB. Furthermore, the Arp2 gene, which is involved in cell protrusion formation, showed about three times more expression at mRNA level, in cells seeding on PDGF-BB-containing scaffold compared to cells seeding on scaffold containing only chitosan nanoparticles, using Real Time PCR test. Finally, under agarose migration assay results demonstrated that cells' chemotaxic behavior was more toward scaffold containing PDGF-BB compared to the PDGF-BB alone or FBS group. In addition, in terms of distance, the cell mass could grow faster, in response to scaffold containing PDGF-BB compared to FBS or PDGF-BB alone; however, the number of migrating cells might be the same or significantly higher in the latter groups.
Assuntos
Becaplermina/farmacologia , Movimento Celular/efeitos dos fármacos , Quitosana/química , Eletricidade , Fibroblastos/citologia , Nanofibras/química , Nanopartículas/química , Bandagens/microbiologia , Becaplermina/química , Proliferação de Células/efeitos dos fármacos , Preparações de Ação Retardada , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Cicatrização/efeitos dos fármacosRESUMO
Over the last few decades, great advancements have been achieved in the field of bone tissue engineering (BTE). Containing a great number of growth factors needed in the process of osteogenesis, platelet rich plasma (PRP) has gained a great deal of attention. However, due to the contradictory results achieved in different studies, its effectiveness remains a mystery. Therefore, in this study, we investigated in vitro performance of co-electrospun PRP/poly ether sulfone/poly(vinyl) alcohol (PRP/PES/PVA) composite scaffolds for the osteogenic differentiation of human adipose-derived mesenchymal stem cells. The activated PRP was mixed with PVA solution to be used alongside PES solution for the electrospinning process. Fourier transform infrared spectroscopy, scanning electron microscopy and tensile tests were performed to evaluate the scaffolds. After confirmation of sustained release of protein, osteogenic potential of the co-electrospun PRP/polymer scaffolds was evaluated by measuring relative gene expression, calcium content, and alkaline phosphatase (ALP) activity. Alizarin red and Hematoxylin and Eosin staining were performed as well. The results of ALP activity and calcium content demonstrated the effectiveness of PRP when combined with PRP-incorporated scaffold in comparison with the other tested groups. In addition, the results of tensile mechanical testing indicated that addition of PRP improves the mechanical properties. Taking these results into account, it appears PES/PVA/PRP scaffold treated with PRP 5% enhances osteogenic differentiation most. In conclusion, incorporation of PRP into electrospun PES/PVA scaffold in this study had a positive influence on osteogenic differentiation of AdMSCs, and thus it may have great potential for BTE applications.
Assuntos
Células-Tronco Mesenquimais/citologia , Nanofibras , Osteogênese , Engenharia Tecidual/métodos , Alicerces Teciduais , Diferenciação Celular , Humanos , Nanofibras/química , Plasma Rico em Plaquetas , Polímeros/química , Álcool de Polivinil/química , Sulfonas/química , Alicerces Teciduais/químicaRESUMO
Electrospray ionization is a wide spread technique for producing polymeric microcarriers (MCs) by applying electrostatic force and ionic cross-linker, simultaneously. In this study, fabrication process of gelatin-chitosan MCs and its optimization using the Response Surface Methodology (RSM) is reported. Gelatin/chitosan (G/C) blend ratio, applied voltage and feeding flow rate, their individual and interaction effects on the diameter and mechanical strength of the MCs were investigated. The obtained models for diameter and mechanical strength of MCs have a quadratic relationship with G/C blend ratio, applied voltage and feeding flow rate. Using the desirability curve, optimized G/C blend ratios that are introduced, include the desirable quantities for MCs diameter and mechanical strength. MCs of the same desirable diameter (350µm) and different G/C blend ratio (1, 2, and 3) were fabricated and their elasticity was investigated via Atomic Force Microscopy (AFM). The biocompatibility of the MCs was evaluated using MTT assay. The results showed that human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) could attach and proliferate on fabricated MCs during 7days of culturing especially on those prepared with G/C blend ratios of 1 and 2. Such gelatin-chitosan MCs may be considered as a promising candidate for injectable tissue engineering scaffolds, supporting attachment and proliferation of hUCMSCs.