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1.
Br J Cancer ; 130(2): 184-200, 2024 02.
Artigo em Inglês | MEDLINE | ID: mdl-38012382

RESUMO

BACKGROUND: The eukaryotic elongation factor, EEF1A2, has been identified as an oncogene in various solid tumors. Here, we have identified a novel function of EEF1A2 in angiogenesis. METHODS: Chick chorioallantoic membrane, tubulogenesis, aortic ring, Matrigel plug, and skin wound healing assays established EEF1A2's role in angiogenesis. RESULT: Higher EEF1A2 levels in breast cancer cells enhanced cell growth, movement, blood vessel function, and tubule formation in HUVECs, as confirmed by ex-ovo and in-vivo tests. The overexpression of EEF1A2 could be counteracted by Plitidepsin. Under normoxic conditions, EEF1A2 triggered HIF1A expression via ERK-Myc and mTOR signaling in TNBC and ER/PR positive cells. Hypoxia induced the expression of EEF1A2, leading to a positive feedback loop between EEF1A2 and HIF1A. Luciferase assay and EMSA confirmed HIF1A binding on the EEF1A2 promoter, which induced its transcription. RT-PCR and polysome profiling validated that EEF1A2 affected VEGF transcription and translation positively. This led to increased VEGF release from breast cancer cells, activating ERK and PI3K-AKT signaling in endothelial cells. Breast cancer tissues with elevated EEF1A2 showed higher microvessel density. CONCLUSION: EEF1A2 exhibits angiogenic potential in both normoxic and hypoxic conditions, underscoring its dual role in promoting EMT and angiogenesis, rendering it a promising target for cancer therapy.


Assuntos
Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Retroalimentação , Fosfatidilinositol 3-Quinases/metabolismo , Células Endoteliais/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Angiogênese , Hipóxia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Fator 1 de Elongação de Peptídeos/genética , Fator 1 de Elongação de Peptídeos/metabolismo
2.
J Cell Biochem ; 123(11): 1793-1807, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-35926149

RESUMO

The dysregulated energy metabolism in white adipose tissues results in derangement of biological signaling resulting in obesity. Lack of vascularization in these white adipose tissues is one of the major reasons for dysregulated energy metabolism. Not much work has been done in this direction to understand the role of angiogenesis in white adipose tissue metabolism. In the present study, we evaluated the effect of angiogenic modulator in the metabolism of white adipocyte (WAC). Bioactive Apigenin was selected and its angiogenic ability was studied. Apigenin was shown to be highly proangiogenic hence the effect of Apigenin on de novo and trans-differentiation of WAT was studied. Apigenin showed enhanced de novo differentiation and trans-differentiation of mouse WAC into brown-like phenotype. To understand the effect of Apigenin on adipose tissue vasculature, coculture studies were conducted. Cross talk between endothelial cell and adipocytes were observed in coculture studies. Gene expression studies of cocultured cells revealed that browning of WAC occurred by triggering the expression of Vascular endothelial growth factor A. The study provides a new insight for inducing metabolic shift in WACs by modulation of angiogenesis in WAC microenvironment by the upregulation of PRDM16 cascade to trigger browning for the treatment of obesity.


Assuntos
Adipócitos Marrons , Fator A de Crescimento do Endotélio Vascular , Camundongos , Animais , Adipócitos Marrons/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Apigenina/farmacologia , Tecido Adiposo Marrom/metabolismo , Tecido Adiposo Branco/metabolismo , Adipócitos Brancos/metabolismo , Fatores de Transcrição/genética , Obesidade/metabolismo , Proteínas de Ligação a DNA/genética
3.
Biotechnol Lett ; 43(10): 2067-2083, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34499291

RESUMO

OBJECTIVE: The study is aimed to verify the possibility of using humanized alternatives to fetal bovine serum (FBS) such as umbilical cord blood plasma (CBP) and AB+ plasma to support the long-term growth of mesenchymal stromal cells (MSCs) derived from the umbilical cord. We hypothesized that umbilical CBP would be a potential substitute to FBS, especially for small scale autologous clinical transplantations. METHODS: The MSCs were cultured for six consecutive passages to evaluate xeno-free media's ability to support long-term growth. Cell proliferation rates, colony-forming-unit (CFU) efficiency and population doublings of expanded MSCs, were investigated. Ex vivo expanded MSCs were further characterized using flow cytometry and quantitative PCR. The impact of cryopreservation and composition of cryomedium on phenotype, viability of MSC was also assessed. RESULTS: Our results on cell proliferation, colony-forming unit efficiency suggested that the expansion of the cells was successfully carried out in media supplemented with humanized alternatives. MSCs showed lower CFU counts in FBS (~ 25) than humanized alternatives (~ 35). The gene expression analysis revealed that transcripts showed significant differential expression by two to three folds in the FBS group compared with MSCs grown in medium with humanized alternatives (p < 0.05). In addition, MSCs grown in a medium with FBS had more osteogenic activity, a signature of unwanted differentiation. The majority of ex vivo expanded MSCs at early and late passages expressed CD44+, CD73+, CD105+, CD90+, and CD166+ in all the experimental groups tested (~ 90%). In contrast to the other MSC surface markers, expression levels of STRO-1+ (~ 21-10%) and TNAP+ (~ 29-11%) decreased with the increase in passage number for MSCs cultured in a FBS-supplemented medium (p < 0.05). CONCLUSION: Our results established that CBP supported culture of umbilical cord tissue-derived MSCs and is a safer Xeno free replacement to FBS. The use of CBP also enables the storage of umbilical cord tissue derived MSCs in patient-specific conditions to minimize adverse events if cells are delivered directly to the patient.


Assuntos
Técnicas de Cultura de Células/métodos , Meios de Cultura/farmacologia , Sangue Fetal/química , Células-Tronco Mesenquimais , Cordão Umbilical/citologia , Animais , Bovinos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Meios de Cultura/química , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Soroalbumina Bovina/farmacologia
4.
Cell Tissue Res ; 375(2): 437-449, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30284087

RESUMO

A xeno-free method for ex vivo generation of red blood cells (RBCs) is attempted in order to replicate for large-scale production and clinical applications. An efficient milieu was formulated using injectable drugs substituting the animal-derived components in the culture medium. Unfractionated mononuclear cells isolated from human umbilical cord blood were used hypothesizing that the heterogeneous cell population could effectively contribute to erythroid cell generation. The strategy adopted includes a combination of erythropoietin and other injectable drugs under low oxygen levels, which resulted in an increase in the number of mature RBCs produced in vitro. The novelty in this study is the addition of supplements to the medium in a stage-specific manner for the differentiation of unfractionated umbilical cord blood mononuclear cells (MNCs) into erythropoietic lineage. The erythropoietic lineage was well established by day 21, wherein the mean cell count of RBCs was found to be 21.36 ± 0.9 × 108 and further confirmed by an upregulated expression of CD235a+ specific to RBCs. The rationale was to have a simple method to produce erythroid cells from umbilical cord blood isolates in vitro by mitigating the effects of multiple erythroid-activating agents and batch to batch variability.


Assuntos
Técnicas de Cultura de Células/métodos , Eritrócitos/citologia , Sangue Fetal/citologia , Leucócitos Mononucleares/citologia , Células Cultivadas , Contagem de Eritrócitos , Feminino , Humanos , Imunofenotipagem , Oxigênio/metabolismo , Preservação Biológica
5.
J Mater Sci Mater Med ; 30(12): 129, 2019 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-31776679

RESUMO

In this study, gelatin-polyethylenimine blend nanofibers (GEL/PEI) were fabricated via electrospinning with different ratios (9:1, 6:1, 3:1) to integrate the properties of both the polymers for evaluating its biomedical application. From scanning electron microscopy, the average diameter of blend nanofibers (265 ± 0.074 nm to 340 ± 0.088 nm) was observed to be less than GEL nanofibers (403 ± 0.08 nm). The incorporation of PEI with gelatin resulted in improved thermal stability of nanofibers whereas the Young's modulus was observed to be higher at 9:1 ratio when compared with other ratios. The in vitro studies showed that the GEL/PEI nanofibers with 9:1 ratio promoted better cell adhesion and viability. GEL/PEI nanofibers with 9:1 and 6:1 showed hemolysis within the permissible limits. From the results, it could be interpreted that GEL/PEI nanofibers with 9:1 ratio proved to be a better scaffold thereby making them a potential candidate for tissue engineering applications.


Assuntos
Gelatina/química , Nanofibras/química , Polietilenoimina/química , Alicerces Teciduais , Materiais Biocompatíveis , Adesão Celular , Linhagem Celular , Proliferação de Células , Humanos , Queratinócitos , Teste de Materiais , Engenharia Tecidual
6.
AAPS PharmSciTech ; 20(3): 110, 2019 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-30756201

RESUMO

Angiogenesis is a vital component of the orchestrated wound healing cascade and tissue regeneration process, which has a therapeutic prominence in treatment of ischemic vascular diseases and certain cardiac conditions. Based on its eminence, several strategies using growth factors have been studied to initiate angiogenesis. However, growth factors are expensive and have short half-life. In this work, sustained release of triiodothyronine, which plays a crucial role in stimulating growth factors and other signaling pathways that are instrumental in initiating angiogenesis, has been attempted through electrospun polycaprolactone nanofibers. This delivery system enabled the slow and sustained delivery of triiodothyronine into the micro-environment, reducing seepage of excess into systemic circulation and eliminating the necessity of repeated dosage forms. It was observed that triiodothyronine-incorporated nanofibers exhibited favorable interaction with cells (phalloidin staining of actin filaments) and also enhanced the rate of endothelial proliferation, migration, and adhesion. The angiogenic potential of these nanofibers was further corroborated through chorioallantoic membrane and rat aortic ring assay (demonstrating cell sprouting area of 3.3 ± 0.71 mm2 compared to 1.2 ± 0.01 mm2 in control). The nanofiber matrix thus fabricated demonstrated a vibrant therapeutic potential to induce angiogenesis. Triiodothyronine also plays a significant role in wound healing independent of initiating angiogenesis. This further substantiates the positive impact of this delivery system as a dressing material for chronic wound therapeutics, ischemic vascular diseases, and certain cardiac conditions.


Assuntos
Portadores de Fármacos , Nanofibras , Neovascularização Fisiológica/efeitos dos fármacos , Tri-Iodotironina/administração & dosagem , Animais , Bandagens , Materiais Biocompatíveis/administração & dosagem , Células Endoteliais da Veia Umbilical Humana , Humanos , Poliésteres/administração & dosagem , Ratos , Tri-Iodotironina/farmacologia , Cicatrização/efeitos dos fármacos
7.
J Mater Sci Mater Med ; 26(2): 101, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25655500

RESUMO

In this study, zein nanofibers based siRNA delivery system has been attempted for the first time. Here, the amphiphilic property of zein and the size advantage of nanofibers have been brought together in developing an ideal delivery system for siRNA. The morphological analysis of the GAPDH-siRNA loaded zein nanofibers revealed the proper encapsulation of the siRNA in the polymeric matrix. The loading efficiency of this delivery system was found to be 58.57±2.4% (w/w). The agarose gel analysis revealed that the zein nanofibers preserved the integrity of siRNA for a longer period even at the room temperature. The in vitro release studies not only depicted the sustaining potential of the zein nanofibers but also ensured the release of sufficient quantity of siRNA required to induce the gene silencing effect. The amphiphilic property of zein supported the cell attachment and thereby facilitated the transfection of siRNA into the cells. qRT-PCR analysis confirmed the potential of the developed system in inducing the desired gene silencing effect. Thus, electrospun zein nanofibers have been successfully employed for the delivery of siRNA which has a great therapeutic potential.


Assuntos
Preparações de Ação Retardada/química , Nanofibras/química , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/genética , Transfecção/métodos , Zeína/química , Células Cultivadas , Preparações de Ação Retardada/administração & dosagem , Galvanoplastia/métodos , Fibroblastos/citologia , Fibroblastos/fisiologia , Inativação Gênica , Humanos , Masculino , Teste de Materiais , Nanocápsulas/química , Nanocápsulas/ultraestrutura , Nanofibras/ultraestrutura , Tamanho da Partícula , Rotação , Resultado do Tratamento
8.
Cell Tissue Res ; 358(3): 833-41, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25260909

RESUMO

Chronic cutaneous wound (CCW) is a major health care burden wherein the healing process is slow or rather static resulting in anatomical and functional restriction of the damaged tissue. Dysregulated expression and degradation of matrix proteins, growth factors and cytokines contribute to the disrupted and uncoordinated healing process of CCW. Therefore, therapeutic approaches for effective management of CCW should be focused towards identifying and manipulating the molecular defects, such as reduced bioavailability of the pro-healing molecules and elevated activity of proteases. This study essentially deals with assessing the expression and integrity of an extracellular matrix protein, Dermatopontin (DPT), in CCW using real-time quantitative reverse transcriptase PCR and immunological techniques. The results indicate that, despite DPT's high mRNA expression, the protein levels are markedly reduced in both CCW tissue and its exudate. To elucidate the cause for this contradiction in mRNA and protein levels, the stability of DPT is analyzed in the presence of wound exudates and various proteases that are naturally elevated in CCW. DPT was observed to be degraded at higher rates when incubated with certain recombinant proteases or chronic wound exudate. In conclusion, the susceptibility of DPT protein to specific proteases present at high levels in the wound milieu resulted in the degradation of DPT, thus leading to impaired healing response in CCW.


Assuntos
Proteoglicanas de Sulfatos de Condroitina/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Pele/metabolismo , Pele/patologia , Cicatrização , Ferimentos e Lesões/metabolismo , Ferimentos e Lesões/patologia , Sequência de Aminoácidos , Proteoglicanas de Sulfatos de Condroitina/química , Proteoglicanas de Sulfatos de Condroitina/genética , Doença Crônica , Eletroforese em Gel de Poliacrilamida , Proteínas da Matriz Extracelular/química , Proteínas da Matriz Extracelular/genética , Exsudatos e Transudatos/metabolismo , Feminino , Gelatina/metabolismo , Humanos , Masculino , Metaloproteinases da Matriz/metabolismo , Pessoa de Meia-Idade , Dados de Sequência Molecular , Proteólise , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Tripsina/metabolismo , Regulação para Cima/genética , Cicatrização/genética , Ferimentos e Lesões/genética
9.
Analyst ; 139(24): 6352-6, 2014 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-25340936

RESUMO

A naphthalimide based fluorescent probe '1' that operates based on photoinduced electron transfer phenomenon is synthesized and its chemosensory application is explored. Among various metal ions, 1 selectively detects Fe(3+) with a detection limit of 3.0 × 10(-8) M. 1 is stable at physiological pH, nontoxic under experimental conditions and suitable for the detection of Fe(3+) ions present in aqueous samples and live cells.


Assuntos
Compostos Férricos/análise , Corantes Fluorescentes/química , Naftalimidas/química , Cátions/análise , Linhagem Celular , Transporte de Elétrons , Humanos , Ferro/análise , Limite de Detecção , Pulmão/citologia , Microscopia de Fluorescência , Modelos Moleculares
10.
AAPS PharmSciTech ; 15(5): 1172-80, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24875151

RESUMO

In this study, we have evaluated the interactions of zein microspheres with different class of drugs (hydrophobic, hydrophilic, and amphiphilic) using in vitro and in silico analysis. Zein microspheres loaded with aceclofenac, metformin, and promethazine has been developed by solvent evaporation technique and analyzed for its compatibility. The physical characterization depicted the proper encapsulation of hydrophobic drug in the microspheres. The in vitro release study revealed the sustaining ability of the microspheres in the following order: hydrophobic > hydrophilic > amphiphilic. In silico analysis also confirmed the better binding affinity and greater interactions of hydrophobic drug with zein. The above results revealed that zein is more suitable for hydrophobic drugs in the development of sustained drug delivery systems using solvent evaporation technique. The study therefore envisages a scope for identifying the most suitable polymer for a sustained drug delivery system in accordance with the nature of the drug.


Assuntos
Microesferas , Preparações Farmacêuticas/química , Zeína/química , Sequência de Aminoácidos , Domínio Catalítico , Química Farmacêutica , Simulação por Computador , Diclofenaco/análogos & derivados , Diclofenaco/química , Composição de Medicamentos , Ligantes , Metformina/química , Modelos Moleculares , Tamanho da Partícula , Prometazina/química , Receptores de Droga/química
11.
Analyst ; 138(4): 1130-6, 2013 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-23254200

RESUMO

Two new rhodamine­indole conjugates 1 and 2 were synthesized for studying their ability to probe specific metal ions at physiological pH in the presence of other competing metal ions. In non-aqueous medium, probe 1 predominantly binds Cu(2+) ions with considerable interference from Fe(3+) and Ni(2+) ions. However, in aqueous medium, 1 exhibits a higher degree of selectivity for Cu(2+) ions with little interference from Fe(3+) ions, and no interference from Ni(2+) ions. The probe 2, obtained by tethering a triazole moiety with probe 1, selectively binds Cu(2+) ions in aqueous medium even in the presence of other metal ions including Fe(3+) ions. The sensitivity of probe 2 to pH variation and interaction with various amino acids were also investigated. The excellent stability of 2 in physiological pH conditions, non-toxicity, non-interference of amino acids in the detection process, and the remarkable selectivity for Cu(2+) ions in aqueous medium allowed the use of 2 in the imaging of live fibroblast cells treated with Cu(2+).


Assuntos
Sobrevivência Celular , Cobre/análise , Solventes/química , Água/análise , Animais , Cobre/química , Camundongos , Células NIH 3T3 , Rodaminas/química , Água/química
12.
AAPS PharmSciTech ; 13(1): 143-9, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22167417

RESUMO

Nonsteroidal anti-inflammatory drugs (NSAIDs) induce gastric injury on long-term usage. This study aims at reducing the side effect of NSAIDs by encapsulating in zein, an acid-resistant biopolymer. Aceclofenac-loaded zein microspheres were prepared by emulsification and solvent evaporation method. The stability of zein microspheres at gastric pH retarded the release of the entrapped drug and hence reduces the possibility of gastric injury. However, the in vitro release of aceclofenac was sustained up to 72 h at intestinal pH. Thus, zein microspheres pave the way for the development of safe and sustained delivery system for NSAIDs thereby achieving the desired therapeutic potential with reduced side effects for chronic inflammatory disorders.


Assuntos
Anti-Inflamatórios não Esteroides/administração & dosagem , Diclofenaco/análogos & derivados , Portadores de Fármacos/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Microesferas , Zeína/administração & dosagem , Animais , Anti-Inflamatórios não Esteroides/síntese química , Anti-Inflamatórios não Esteroides/metabolismo , Células Cultivadas , Preparações de Ação Retardada , Diclofenaco/administração & dosagem , Diclofenaco/síntese química , Diclofenaco/metabolismo , Portadores de Fármacos/síntese química , Portadores de Fármacos/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Zeína/síntese química , Zeína/metabolismo
13.
Colloids Surf B Biointerfaces ; 216: 112589, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35660195

RESUMO

Rare earth lanthanum oxide nanoparticle reinforced collagen biomatrix that elicited the endothelial cell activation to promote angiogenesis for biomaterial integration was developed and evaluated in the present study. The structural integrity of collagen was not compromised on crosslinking of lanthanum oxide nanoparticle to collagen biomolecule. As-synthesised collagen biomatrix was shown to have improved mechanical strength, a lesser susceptibility to proteolytic degradation and good swelling properties. Superior cytocompatibility, hemocompatibility and minimal ROS generation was observed with Lanthanum oxide nanoparticle reinforced collagen bio matrix. The Lanthanum oxide nanoparticle reinforced collagen bio matrix elicited endothelial cell activation eliciting pro-angiogensis as observed in tube formation and aortic arch assays. The bio-matrix promoted the infiltration and proliferation of endothelial cells which is an unexplored domain in the area of tissue engineering that is very essential for biomaterial integration into host tissue. The wound healing effect of Lanthanum oxide nanoparticle stabilized collagen showed enhanced cell migration in vitro in cells maintained in Lanthanum oxide nanoparticle reinforced collagen bio matrix. The study paves the way for developing rare earth-based dressing materials which promoted biomatrix integration by enhancing vascularisation for tissue regenerative applications in comparison with traditional biomaterials.


Assuntos
Materiais Biocompatíveis , Nanopartículas , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Colágeno/química , Colágeno/farmacologia , Células Endoteliais , Lantânio , Óxidos , Alicerces Teciduais/química
14.
Neurochem Int ; 159: 105386, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35803325

RESUMO

BACKGROUND: Scaffold proteins coordinate multiple signalling pathways by integrating various proteins but the role of these proteins in neuronal pathways remains to be elucidated. The present study focused to evaluate the expression of the scaffold protein CULLIN2 in neuronal cells. METHODS: The neuronal precursor cell line N2A was differentiated to neurons in-vitro with retinoic acid and biochemical assays were used to understand the gene expression profiling of CULLIN2. Moreover, neddylation inhibitor MLN4924 was used to inhibit the activity of CULLIN2 and the downstream substrates were validated. Finally, the role of CULLIN2 in nerve regeneration was evaluated in an in vivo zebrafish model. RESULTS: Experimental data showed that the neuronal cells N2A have lower expression of CULLIN2 compared to skin cell lines (HaCaT and A431) and inactivation with the neddylation inhibitor resulted in cell death. Furthermore differentiating the neural precursor cell line into neurons with retinoic acid enhanced the expression of CULLIN2. Examining downstream signalling molecules with the neddylation inhibitor illuminates that MLN4924 treatment influences the cytokine signalling cascade (JAK-STAT) in neuronal cells. Moreover, for the first time, we show that the ubiquitin ligase protein CULLIN2 is perturbed in neural regeneration. Expression profile of CULLIN2 was significantly decreased in response to a nerve injury in Zebra fish and as the nerve regenerates there is corresponding reduction in the mRNA levels. CONCLUSION: During differentiation CULLIN2 is upregulated whereas during regeneration there is significant downregulation. Thus, our findings reveal a crucial role of the scaffold protein CULLIN2 in nerve differentiation and regeneration which can be vital for the treatment of nerve injury.


Assuntos
Transdução de Sinais , Peixe-Zebra , Animais , Diferenciação Celular , Regeneração Nervosa/fisiologia , Neurônios , Tretinoína/farmacologia
15.
J Trace Elem Med Biol ; 72: 126983, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35537228

RESUMO

INTRODUCTION: The highly complex pathophysiology of the wound micro-environment demands the development of a multi-faceted system which would enhance the wound healing cascade. Incorporation of nanotechnology in wound therapeutics has opened up new avenues to tourment the diseased condition. Amongst the various types of nanoparticles molybdenum oxide nanoparticles posses various inherent properties that makes it a versatile material to be used in healing. Incorporation of Molybdenum nanoparticles into collagen scaffolds would provide a synergistic and sequential healing process ensuring the formation of a fully functional tissue. MATERIALS AND METHODS: The physico-chemical characterization of the synthesized materials were done using SEM and FT-IR techniques. The bicompatibility and cell proliferation were tested using HaCaT cell lines. Pro-angiogenic ability of the scaffold was tested using CAM assay and Chick aortic arch assay. Finally the in-vivo wound healing ability of the material was tested by creating wound of about 6 cm2 on the dorsal side of Wistar rats and observed for about 21 days. RESULTS: The characterization of the scaffold revealed the presence MoO3 nanoparticles and their structural integrity within the scaffold. The synthesized MoO3-collagen nanocomposite was found to be biocompatible and hemocompatible. The in-vitro studies demonstrated that the MoO3-collagen scaffold significantly increased the cell adhesion and migration to nearly 2 fold. The MoO3 embedded collagen sheets synergistically favoured neovascularization and re-epithelization,which would potentially enhance therapeutic efficiency of the scaffold. The nanocomposite also encouraged results in in-vivo analysis, the Wistar rats treated with MoO3-collagen scaffolds showed complete healing in about 15 days. CONCLUSION: The fabricated MoO3-collagen scaffold was found to play an important role in all major events of wound healing such as adhesion, migration, proliferation and angiogenesis. The in-vivo healing assay also proved that the healing rate of animals treated with the samples was comparatively faster. Further research using various trace elements would open up promising avenues in healing therapeutics.


Assuntos
Molibdênio , Nanopartículas , Animais , Colágeno , Nanopartículas/química , Óxidos/farmacologia , Ratos , Ratos Wistar , Espectroscopia de Infravermelho com Transformada de Fourier , Alicerces Teciduais/química
16.
Int J Biol Macromol ; 219: 907-918, 2022 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-35952816

RESUMO

Scars occur as a result of fibrosis after tissue damage or surgery and reports suggest that excessive Transforming growth factor-ß (TGF-ß) activity during the process of wound healing leads to progressive fibrosis. Decorin is an extracellular matrix (ECM) protein which regulates collagen fibrillogenesis. However, targeted delivery and effective protein therapy remains a challenge owing to degradation byproteases. Hence, we aimed to deliver Decorin in a sustainable mode for the reduction of TGF-ß levels and subsequent scar formation. Herein, we have fabricated PCL-Gelatin bio-mimetic scaffolds to optimize the bio-activity and provide localized delivery of recombinant Decorin. The degradation and drug release patterns reveals that this biomaterial is biodegradable and offers sustained release of the recombinant Decorin. Decorin loaded nanofiber displayed lower adhesion and proliferation rates in in-vitro conditions. Moreover, Decorin loaded scaffolds demonstrated morphological changes in cells, specifically targeting the myofibroblast. The expression of TGF-ß was also scrutinized to understand the effect of Decorin loaded nanofibers. Besides, in the in-vitro fibrotic model, Decorin loaded nanofibers efficiently reduced the expression of ECM related proteins. Therefore, we report the sustained delivery of the recombinant Decorin from nanofiber dressing to potentially obstruct scar formation during the process of wound healing.


Assuntos
Cicatriz , Gelatina , Materiais Biocompatíveis/uso terapêutico , Biomimética , Cicatriz/tratamento farmacológico , Cicatriz/patologia , Colágeno/metabolismo , Decorina , Preparações de Ação Retardada/uso terapêutico , Proteínas da Matriz Extracelular , Fibrose , Humanos , Fator de Crescimento Transformador beta , Fator de Crescimento Transformador beta1/metabolismo , Fatores de Crescimento Transformadores/uso terapêutico
17.
Int J Biol Macromol ; 166: 333-341, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33122062

RESUMO

Wound healing is a complex process which requires appropriate structural support for restoration of tissue continuity and function. Collagen can act as a template for cellular activities but poor physico-chemical properties necessitates the stabilization of collagen without impairing its structure and function. This study investigates the effect of magnesium ascorbyl phosphate (MAP) on collagen with reference to physico-chemical properties. Incorporation of MAP enhanced the rate of collagen fibrillation signifying increased interaction at reduced time interval. MAP did not induce any changes in the secondary structure of collagen while there was an increase in shear viscosity with increase in shear stress at different shear rate. MAP stabilized collagen film exhibited higher denaturation temperature and showed an increase in Young's Modulus when compared with that of collagen film. In vivo studies showed complete wound closure on day 16 in case of stabilized collagen film. Mechanical properties of healed skin revealed that MAP collagen film treated rat skin completely regained its properties similar to that of normal skin thereby making them a potential candidate for wound healing application.


Assuntos
Ácido Ascórbico/análogos & derivados , Colágeno/química , Cicatrização , Animais , Ácido Ascórbico/química , Ácido Ascórbico/farmacologia , Bandagens , Colágeno/metabolismo , Módulo de Elasticidade , Feminino , Células HaCaT , Humanos , Camundongos , Multimerização Proteica , Estabilidade Proteica/efeitos dos fármacos , Ratos , Ratos Wistar
18.
Eur J Pharm Sci ; 167: 106021, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34571179

RESUMO

Electrospinning, a recent fast-emerging technique highly applicable in the production of nanofibers has gained vast recognition owing to its explicit applications in various domains. Amongst which, the production of nanoscaffolds for wound healing applications has been focused recently due to advantages over conventional wound healing methods. In the present research, a composite nanoscaffold comprising SBA-15 (Santa Barbara Amorphous), amine functionalized SBA-15 polycaprolactone (PCL) and curcumin was investigated for its potentiality in wound healing therapeutics. The high biocompatibility and cell adhesion of amine functionalized SBA-15 and the widely explored antimicrobial properties of curcumin added benefit for the wound healing target. The prepared highly interconnected electrospun fibers with porous structure were characterized through various studies such as FTIR, XRD, SEM and EDAX. Further, antibacterial studies against both Gram positive (Bacillus subtilis) and Gram negative (Escherichia coli) strains revealed an improved zone of inhibition. Major invitro studies such as cell migration, proliferation, bio-compatibility was experimented through cell adhesion and live and dead assay using Swiss 3T6 cell lines. In vivo studies on female Wister rats using the fabricated nanofibers incorporated with curcumin and amine functionalized SBA-15 showed 99% scar-less wound healing within 21 days. Re-epithelization of tissue, collagen deposition and formation of granulation tissue were observed from the results of Hematoxylin-Eosin and Masson's tri-chrome staining. From the observations, it can be concluded that the fabricated nanoscaffold could be an effective substrate for wound healing therapeutics.


Assuntos
Curcumina , Nanofibras , Animais , Feminino , Ratos , Ratos Wistar , Dióxido de Silício , Cicatrização
19.
Biol Trace Elem Res ; 194(1): 121-134, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31123924

RESUMO

Conventional cancer therapies possess a plethora of limitations which led to the awakening of nanotechnology and nanomedicine. However, technological success is widely dependent on complete understanding of the complexity and heterogeneity of tumor biology on one hand and nanobiointeractions associated with challenges of synthesis, translation, and commercialization on the other. The present study therefore deals with one such targeted approach aiming at synthesizing, characterizing, and understanding the efficacy of molybdenum oxide nanoparticles. The phase structure, morphology, and elemental composition of the synthesized nanoparticles were characterized using Fourier transform infrared spectroscopy, Raman spectroscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and scanning electron microscopy. The cytotoxicity studies revealed that the IC50 vales of molybdenum trioxide (MoO3) particles against skin cancer cells (melanoma and non-melanoma) were around 200-300 µg. The nanoparticles were found to induce mitochondrial-mediated apoptosis driven by the apoptotic genes such as BAX and Bcl2. Molybdenum being a cofactor for the majority of metabolic enzymes could have triggered the selective internalization of the nanoparticles which in turn could have modified the granularity of the cytoplasm and subsequently lead to mitochondrial-mediated apoptosis. Further, the anti-angiogenic property of MoO3 nanoparticles was corroborated using Chick chorioallantoic membrane (CAM) assay and aortic ring assay. Taken together , unraveling the role of MoO3 nanoparticles in cancer and angiogenesis opens up venues for nano biological intervention of selective cancer cell targeting with minimal damage to the normal cells using natural trace elements that are generally known to influence various metabolic enzymes.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Melanoma/tratamento farmacológico , Molibdênio/farmacologia , Nanopartículas/química , Óxidos/farmacologia , Neoplasias Cutâneas/tratamento farmacológico , Células 3T3 , Animais , Antineoplásicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Melanoma/metabolismo , Melanoma/patologia , Camundongos , Mitocôndrias/efeitos dos fármacos , Molibdênio/química , Neovascularização Patológica/tratamento farmacológico , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Óxidos/química , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia
20.
Biomed Mater ; 15(3): 035009, 2020 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-31935710

RESUMO

Fabricating and designing a scaffold is a complex and highly challenging process in the current scenario. The present study deals with the design and fabrication of electrospun Santa Barbara Amorphous (SBA)-15-incorporated polyvinyl alcohol (PVA) with curcumin, which can be used as a biomimetic nanoscaffold for skin tissue engineering. Curcumin was selected due to its effective anti-microbial and anti-inflammatory properties. SBA-15 was selected for its characteristic drug-carrying potential. Fourier transform infrared spectroscopy and x-ray diffraction characterizations of the fabricated nanofiber demonstrated the interaction of PVA, SBA-15 and curcumin. The scanning electron microscopy results depicted that the nanofiber was highly interconnected with a porous structure mimicking the extracellular matrix. The nanofibrous scaffold showed a higher percentage of cell migration, proliferation, cytocompatibility and biocompatibility with absence of cytotoxicity which was evidenced from the results of MTT assay, cell adhesion and live/dead assay using HaCaT cells. The results of the anti-bacterial test depicted that the synthesized nanofiber forms a potent material for skin wound-healing therapeutics. The in vitro drug release study performed over a period of 80 h revealed a sustained release pattern of curcumin from the SBA-15-incorporated PVA nanofiber. Finally, the in vivo results confirmed that SBA-15-incorporated PVA nanofiber with curcumin showed efficient wound-healing activities.


Assuntos
Biomimética , Curcumina/química , Álcool de Polivinil/química , Dióxido de Silício/química , Pele/efeitos dos fármacos , Engenharia Tecidual/métodos , Adsorção , Animais , Antibacterianos/química , Anti-Infecciosos/química , Anti-Inflamatórios/química , Materiais Biocompatíveis/química , Adesão Celular , Linhagem Celular , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Feminino , Humanos , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Nanofibras/química , Nitrogênio/química , Ratos , Ratos Wistar , Espectroscopia de Infravermelho com Transformada de Fourier , Alicerces Teciduais , Cicatrização , Difração de Raios X
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