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1.
Microb Pathog ; 135: 103634, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31325568

RESUMO

BACKGROUND: Research indicates that Helicobacter pylori can inflict severe histological damage through the modulation of host-related genes. The current study investigated the effect of H. pylori genotypes in the outcome of disease, and the expression of anti-apoptotic related genes, COX-1, COX-2, and iNOS genes in benign, pre-malignant, and malignant lesions of gastric carcinogenesis. MATERIALS AND METHODS: Tissue samples from H. pylori positive patients were graded based on the genotype of the infected H. pylori strain. Expression of COX-1, COX-2 and iNOS was assessed using a combination of real-time PCR and immunohistochemistry. RESULTS: Gene expression studies confirmed that COX-2 and iNOS expression was highly and selectively induced in epithelium with premalignant changes such as atrophic conditions, metaplasia and dysplasia, suggesting an important role of these genes in the sequence to gastric carcinoma of the intestinal type. Furthermore, the expression of COX-2 and iNOS was also dependent on the genotype of H. pylori and subjects with genotype-1 exhibited significantly higher expressions of COX-2 and iNOS compared to other genotypes. Comparison of the expression levels among infected and uninfected individuals demonstrated significant difference in the expression pattern of COX-2 gene whereas iNOS expression was found only in subjects infected H. pylori (p < 0.001). Immunohistochemical staining showed 1.5619 folds higher propensity of COX-2 and 3.2941 folds higher intensity of iNOS expression in subjects infected with H. pylori genotype 1. CONCLUSION: The up-regulation of COX-2 and iNOS was associated with the genotype of the H. pylori strain and the presence of certain genotype may greatly affect early events during carcinogenesis.


Assuntos
Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/metabolismo , Expressão Gênica , Infecções por Helicobacter/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Adenocarcinoma , Adulto , Ciclo-Oxigenase 1/genética , Ciclo-Oxigenase 2/genética , Mucosa Gástrica/patologia , Gastrite , Gastrite Atrófica , Genótipo , Infecções por Helicobacter/genética , Infecções por Helicobacter/patologia , Helicobacter pylori/patogenicidade , Humanos , Imuno-Histoquímica , Metaplasia , Pessoa de Meia-Idade , Óxido Nítrico Sintase Tipo II/genética , RNA Mensageiro/metabolismo , Neoplasias Gástricas/patologia , Regulação para Cima
2.
J Artif Organs ; 17(4): 291-300, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25030000

RESUMO

Demand of donor organs for transplantation in treatment of organ failure is increasing. Hence there is a need to develop new strategies for the alternative sources of organ development. Attempts are being made to use xenogenic organs by genetic manipulation but the organ rejection against human always has been a major challenge for the survival of the graft. Advancement in the genetic bioengineering and combination of different allied sciences for the development of humanized organ system, the therapeutic influence of stem cell fraction on the reconstitution of organ architecture and their regenerative abilities in different tissues and organs provides a better approach to solve the problem of organ shortage. However, the available strategies for generating the organ/tissue scaffolds limit its application due to the absence of complete three-dimensional (3D) organ architecture, mechanical strength, long-term cell survival, and vascularization. Repopulation of whole decellularized organ scaffolds using stem cells has added a new dimension for creating new bioengineered organs. In recent years, several studies have demonstrated the potential application of decellularization and recellularization approach for the development of functional bio-artificial organs. With the help of established procedures for conditioning, extensive stem cells and organ engineering experiments/transplants for the development of humanized organs will allow its preclinical evaluation for organ regeneration before translation to the clinic. This review focuses on the major aspects of organ scaffold generation and repopulation of different types of whole decellularized organ scaffolds using stem cells for the functional benefit and their confines.


Assuntos
Técnicas de Cultura de Órgãos/métodos , Regeneração/fisiologia , Células-Tronco/fisiologia , Engenharia Tecidual/métodos , Alicerces Teciduais , Humanos
3.
Inflamm Res ; 61(4): 359-65, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22193858

RESUMO

OBJECTIVE: The present study was designed to investigate the role of X-ray cross-complementing group 1 (XRCC1) and apurinic/apyrimidinic endonuclease 1 (APE1) polymorphisms in apoptosis and the risk of ulcerative colitis (UC). MATERIALS AND METHODS: Blood samples from 384 unrelated subject (age range 18-65 years; 171 with UC, 213 healthy controls) were collected after colonoscopy. Genomic DNA was isolated and genotyped for XRCC1 Arg399Gln and APE1 Asp148Glu using a confronting two-pair primers polymerase chain reaction. Apoptosis and intracellular reactive oxygen species (ROS) levels in peripheral blood mononuclear cells were measured using annexin-V and H(2)DCFDA assay, respectively. RESULTS: The frequency of genotype Arg399Gln (heterozygous) of XRCC1 gene was significantly higher in patients with UC than the controls (odds ratio [OR] 1.73; 95% confidence interval [CI] 1.13-2.64; p = 0.01). Similarly the genotypic frequency of APE1 Asp148Glu showed statistically significant incidence among UC subjects (OR 1.54; 95% CI 1.02-2.33; p = 0.04). Polymorphism in XRCC1 Arg399Gln and APE1 Asp148Glu together considerably increased the risk of UC (OR 2.303; 95% CI 1.43-3.69; p = 0.0007). ROS levels were high in UC subjects compared with controls (p = 0.01). CONCLUSION: Polymorphisms in XRCC1 Arg399Gln and APE1 Asp148Glu significantly increased the rate of apoptosis and risk of ulcerative colitis.


Assuntos
Apoptose/genética , Colite Ulcerativa/genética , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/genética , Proteínas de Ligação a DNA/genética , Adolescente , Adulto , Idoso , Colite Ulcerativa/metabolismo , Feminino , Predisposição Genética para Doença , Humanos , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Espécies Reativas de Oxigênio/metabolismo , Risco , Proteína 1 Complementadora Cruzada de Reparo de Raio-X , Adulto Jovem
4.
Neurol India ; 70(3): 1102-1111, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35864646

RESUMO

Background and Aim: The major aim of this study was to identify the most common stemness genes across different stem cell types and further validate them in human fetal subventricular zone-derived primary and cultured neural precursor cells (NPCs). This study involved the use of a unique method of stemness meta-analysis (SMA) for investigating comprehensive upregulation and downregulation of differentially expressed genes (DEGs) among different stem cell populations. Materials and Methods: A total of 55 mouse and human data sets targeting crucial genes identified in seven different types of stem cells population were screened and subjected to independent DEGs analysis using SMA. Identified 30 meta-gene signatures were subjected to functional enrichment analysis based on their biological processes and molecular functions. Validation of enriched meta-gene signatures was performed using RT-qPCR. Cellular localization of ABCB1 and ABCG2 was identified using immunofluorescence staining, whereas functional assessment was performed using western-blot. Results: SMA analysis revealed that among 52 commonly expressed genes, 30 genes were either upregulated or downregulated in at least two stem cell populations. Further gene enrichment analysis showed nine genes (ABCB1, ABCG2, HSPA4, HSPA9, HSPA14, Nestin, Sox-2, Oct-4, and Notch-2) with the highest combined scores among 30 meta-gene signatures. RT-qPCR demonstrated that all the enriched gene signatures were significantly upregulated in primary NPCs and further downregulated during NPCs lineage differentiation in culture except HSPA4, HSPA9, and HSPA14 gene transcripts. Conclusions: The stemness meta-gene signatures were abundantly expressed in human NPCs population which categorically suggest the involvement of these genes/pathways in pluripotency maintenance and molecular switches for lineage differentiation while HSP-70 had a neuroprotective effect.


Assuntos
Transportadores de Cassetes de Ligação de ATP , Células-Tronco Neurais , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Diferenciação Celular/genética , Células Cultivadas , Expressão Gênica , Humanos , Camundongos , Células-Tronco Neurais/metabolismo
5.
Cell Mol Bioeng ; 15(1): 137-150, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35096189

RESUMO

INTRODUCTION: Human mesenchymal stem cells (hMSCs) holds great promise for managing several clinical conditions. However, the low engraftment efficiency and obscurity to harvest these cells without compromising the cellular viability, structural and functional properties from the culture niche still remain major obstacles for preparing intact regenerative constructs. Although few studies have demonstrate different methods for generating cell-liberated amniotic scaffolds, a common method for producing completely cell-liberated amnion (D-HAM) and chorion (D-HCM) scaffolds and their cytocompatibility with hMSCs yet to be demonstrated. METHODS: A common process was developed for preparing D-HAM and D-HCM scaffolds for assessing hMSCs engraftment efficiency, proliferation and molecular shifts to generate cell-laden biological discs. The structural and functional integrity of D-HAM and D-HCM was evaluated using different parameters. The compatibility and proliferation efficiency of hMSCs with D-HAM and D-HCM was evaluated. RESULTS: Histological analysis revealed completely nucleic acid-free D-HAM and D-HCM scaffolds with intact extracellular matrix, mechanical and biological properties almost similar to the native membranes. Human MSCs were able to adhere and engraft on D-HCM better than D-HAM and expanded faster. Ultrastructural observations, crystal violet staining and expression studies showed better structural and functional integrity of hMSCs on D-HCM than D-HAM and control conditions. CONCLUSION: A common, simple and reliable process of decellularization can generate large number of cell-liberated amniotic scaffolds in lesser time. D-HCM has better efficiency for hMSCs engraftment and proliferation and can be utilized for preparing suitable cell-laden constructs for tissue engineering applications.

6.
Tissue Cell ; 73: 101631, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34461569

RESUMO

INTRODUCTION: The reconstruction/regeneration of human bone injuries/defects represents a crucial challenge due to the lack of suitable bio/immune compatible and implantable biological grafts. The available strategies represent implications of several types of grafting materials in the form of metals, synthetic, and various kinds of biological scaffolds; however, the lack of appropriate biological components required for activating and enhancing repair mechanisms at the lesion-site limits their wider applicability. METHODS: In this study, a unique approach for generating human osteogenic implantable grafts was developed using biofabrication technology. Using a gradient change of detergents and continuous agitation, developed a unique technique to generate completely cell-free amnion and chorion scaffolds. The absence of cellular components and integrity of biological and mechanical cues within decellularized human amnion (D-HAM) and chorion (D-HCM) were evaluated and compared with fresh membranes. Allogenic bone grafts were prepared through induction of human mesenchymal stem cells (hMSCs) into osteogenic cells on D-HAM and D-HCM and evaluated for their comparative behavior at the cellular, histological and molecular levels. RESULTS: The common decellularization process resulted in an efficient way to generate D-HAM and D-HCM while retaining their intact gross-anatomical architecture, surface morphology, extracellular matrix components, and mechanical properties. Both these scaffolds supported better growth of human umbilical cord blood derived MSCs as well as osteogenic differentiation. Comparative investigation revealed better growth rate and differentiation on D-HCM compared to D-HAM and control conditions. CONCLUSION: D-HCM could be used as a better choice for producing suitable allogenic bone grafts for efficient bone healing applications.


Assuntos
Âmnio/citologia , Transplante Ósseo , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Âmnio/ultraestrutura , Regeneração Óssea , Cálcio/metabolismo , Adesão Celular , Diferenciação Celular , Córion/citologia , Córion/ultraestrutura , Humanos , Imunofenotipagem , Ácidos Nucleicos/metabolismo , Osteogênese , Transplante Homólogo
8.
Inflammopharmacology ; 18(2): 59-64, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20143166

RESUMO

Persistent infection with Helicobacter pylori confers an increased risk of peptic ulceration and gastric adenocarcinoma. Reactive oxygen and nitrogen species play a crucial role in the progression from normal gastric mucosa to cancer. The aim of the present study was to investigate the plasma malondialdehyde and nitric oxide levels in H. pylori related gastroduodenal diseases and associate their levels with gastric pathology and genotypes of H. pylori. Malondialdehyde and nitric oxide levels in plasma samples of 250 subjects were spectrophotometrically determined. Subsequently, genotypic and histopathological assessment was performed in gastric biopsies obtained during endoscopy. The levels of MDA and NO exceeded in subjects infected with genotype-1 of Hp than those with other genotypes suggesting more precise interaction of highly virulent strains of Hp in eliciting severe tissue damage. In conclusion, the study demonstrates close relationship between the plasma malondialdehyde and nitric oxide levels, gastric histopathology and genotypes of H. pylori.


Assuntos
Gastroenteropatias/sangue , Gastroenteropatias/microbiologia , Infecções por Helicobacter/sangue , Helicobacter pylori/genética , Helicobacter pylori/patogenicidade , Malondialdeído/sangue , Óxido Nítrico/sangue , Adulto , Feminino , Gastrite/sangue , Gastrite/microbiologia , Gastrite/patologia , Gastrite Atrófica/sangue , Gastrite Atrófica/microbiologia , Gastrite Atrófica/patologia , Gastroenteropatias/patologia , Genótipo , Infecções por Helicobacter/genética , Infecções por Helicobacter/patologia , Helicobacter pylori/isolamento & purificação , Humanos , Intestinos/microbiologia , Intestinos/patologia , Masculino , Metaplasia/sangue , Metaplasia/microbiologia , Metaplasia/patologia , Pessoa de Meia-Idade , Neoplasias Gástricas/sangue , Neoplasias Gástricas/microbiologia , Neoplasias Gástricas/patologia
10.
Mater Sci Eng C Mater Biol Appl ; 102: 34-44, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31147006

RESUMO

Spinal cord injury (SCI) is one of the most precarious conditions which have been one of the major reasons for continuous increasing mortality rate of SCI patients. Currently, there is no effective treatment modality for SCI patients posing major threat to the scientific and medical community. The available strategies don't mimic with the natural processes of nervous tissues repair/regeneration and majority of the approaches may induce the additional fibrotic or immunological response at the injury site and are not readily available on demand. To overcome these hurdles, we have developed a ready to use bioengineered human functional neurological construct (BHNC) for regenerative applications in SCI defects. We used cryopreserved meningeal tissues (CMT) for bioengineering these neurological constructs using acellularization and repopulation technology. The technology adopted herein generates intact neurological scaffolds from CMT and retains several crucial structural, biochemical and mechanical cues to enhance the regenerative mechanisms. The neurogenic differentiation on CMT scaffolds was almost similar to the freshly prepared meningeal scaffolds and mimics with the natural nervous tissue developmental mechanisms which offer intact 3D-microarchitecture and hospitable microenvironment enriched with several crucial neurotrophins for long-term cell survival and function. Functional assessment of developed BHNC showed highly increased positive staining for pre-synaptic granules of Synapsis-1 along with MAP-2 antibody with punctuate distribution in axonal regions of the neuronal cells which was well supported by the gene expression analysis of functional transcripts. Given the significant improvement in the field may enable to generate more such ready to use functional BHNC for wider applicability in SCI repair/regeneration.


Assuntos
Materiais Biomiméticos/farmacologia , Criopreservação , Meninges/fisiologia , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Fenômenos Biomecânicos , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Humanos , Meninges/efeitos dos fármacos , Meninges/ultraestrutura , Células-Tronco Neurais/citologia , Células-Tronco Neurais/efeitos dos fármacos , Células-Tronco Neurais/ultraestrutura , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
11.
Mater Sci Eng C Mater Biol Appl ; 98: 861-873, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30813092

RESUMO

Acute liver failure (ALF) is one of the most devastating fatal conditions which have posed crucial challenges to the clinicians and researchers for identifying permanent cure. Currently liver transplantation has been considered as the only managerial option. However it's wider applicability has been limited owing to non-availability of quality donor organs, cost-intensiveness, surgical hitches, life-long use of immunosuppressive drugs and long-term complications. Since last decades, several liver support systems have been developed for the management of failing liver in acute condition. However, the major limitation has been the lack of natural biological support and long-term survival of the grafts post-transplantation. Repopulation of decellularized xenogeneic organs is one of the emerging technologies for development of humanized neo-organs for demanding regenerative application. However, the earlier reported studies do not fulfil the insistence to provide immunologically tolerable humanized liver grafts for clinical applications. Here we demonstrate an efficient approach to generate transplantable humanized liver grafts which provides long-term support to the failing liver in Acute Liver Failure (ALF) animal models. These bioengineered humanized liver tissue grafts expresses several liver specific transcripts and performed crucial synthetic (albumin production) and detoxification (urea synthesis) functions at comparative level to normal liver. Intraperitoneal transplantation of these humanized liver grafts offered favourable microenvironment to exchange toxic substances across the barrier during ALF condition and provided long-term survival and function of the graft. In summary, the results of present study provide a first proof of concept in pre-clinical ALF animal model for the applicability of these bioengineered humanized livers in the management of failing liver on demand and may be considered as potential bridge to liver transplantation.


Assuntos
Bioengenharia , Falência Hepática Aguda/terapia , Transplante de Fígado , Peritônio/cirurgia , Animais , Biomarcadores/metabolismo , Movimento Celular , Modelos Animais de Doenças , Regulação da Expressão Gênica , Humanos , Fígado/irrigação sanguínea , Fígado/cirurgia , Fígado/ultraestrutura , Masculino , Imagem Óptica , Ratos Wistar , Recuperação de Função Fisiológica , Esterilização , Alicerces Teciduais/química , Transplante Heterólogo
12.
Gene ; 675: 165-175, 2018 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-30180963

RESUMO

BACKGROUND: The present study has been aimed to identify molecular dynamics of pancreatic transcription factors (pTFs) during events of directed trans-differentiation of human hepatic progenitor cells (hHPCs) into insulin producing cells (InPCs) within bioengineered humanized neoorgan. The study demonstrates applicability of acellularized whole splenic scaffold (ASOS) to generate insulin producing humanized transplantable neoorgan through activation of pancreatic transcription factors. METHODS: An efficient acellularization process was developed for xenogeneic rat spleen using change in different gradients of reagents perfusion through splenic artery for varying time points. The acellularized xenogeneic spleen scaffold was characterized thoroughly for preservation of extra-cellular matrix and retention of organ specific vasculature and mechanical properties. Further scaffolds were sterilized and repopulated with hHPCs which were triggered using a stage wise induction with growth factors and hyperglycemic challenge for trans-differentiation into InPCs. Dynamics of pTFs alone or simultaneously during induction process was identified using gene expression analysis and immunological staining. RESULTS: The cells within the engineered neoorgan respond to growth factors and extrinsic hyperglycemic challenge and generate large number of InPCs under controlled dynamic regulation of pTFs. Highly controlled regulation of pTFs generates higher percentage of Nkx-6.1+/C-peptide+ cells within the engineered splenic scaffolds. Generation of high percentage of insulin and C-peptide positive cells in three-dimensional organ architecture responded better to hyperglycemic stimuli and produced higher quantity of insulin than 2D-culture system. CONCLUSION: The present study provides a novel platform for designing effective regenerative strategies using whole organ scaffolds to control hyperglycemia under tight regulation of pTFs using humanized neoorgan system.


Assuntos
Células Secretoras de Insulina/metabolismo , Insulina/metabolismo , Engenharia Tecidual/métodos , Fatores de Transcrição/metabolismo , Animais , Peptídeo C/genética , Peptídeo C/metabolismo , Diferenciação Celular , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Hiperglicemia/metabolismo , Simulação de Dinâmica Molecular , Ratos , Baço/citologia , Transplante de Células-Tronco , Células-Tronco/citologia , Células-Tronco/metabolismo , Técnicas de Cultura de Tecidos , Alicerces Teciduais , Transativadores/genética , Transativadores/metabolismo , Fatores de Transcrição/genética
13.
World J Hepatol ; 10(11): 822-836, 2018 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-30533183

RESUMO

End stage liver diseases (ESLD) represent a major, neglected global public health crisis which requires an urgent action towards finding a proper cure. Orthotropic liver transplantation has been the only definitive treatment modality for ESLD. However, shortage of donor organs, timely unavailability, post-surgery related complications and financial burden on the patients limits the number of patients receiving the transplants. Since last two decades cell-based therapies have revolutionized the field of organ/tissue regeneration. However providing an alternative organ source to address the donor liver shortage still poses potential challenges. The developments made in this direction provide useful futuristic approaches, which could be translated into pre-clinical and clinical settings targeting appropriate applications in specific disease conditions. Earlier studies have demonstrated the applicability of this particular approach to generate functional organ in rodent system by connecting them with portal and hepatic circulatory networks. However, such strategy requires very high level of surgical expertise and also poses the technical and financial questions towards its future applicability. Hence, alternative sites for generating secondary organs are being tested in several types of disease conditions. Among different sites, omentum has been proved to be more appropriate site for implanting several kinds of functional tissue constructs without eliciting much immunological response. Hence, omentum may be considered as better site for transplanting humanized bioengineered ex vivo generated livers, thereby creating a secondary organ at intra-omental site. However, the expertise for generating such bioengineered organs are limited and only very few centres are involved for investigating the potential use of such implants in clinical practice due to gap between the clinical transplant surgeons and basic scientists working on the concept evolution. Herein we discuss the recent advances and challenges to create functional secondary organs through intra-omental transplantation of ex vivo generated bioengineered humanized livers and their further application in the management of ESLD as a supportive bridge for organ transplantation.

14.
Artigo em Inglês | MEDLINE | ID: mdl-30443545

RESUMO

Spinal cord injury (SCI) is one of the most devastating conditions echoes with inflammation, enhanced fibrosis and larger axonal gaps due to destruction of neurological cells which has caused continuous increasing mortality rate of SCI patients due to absence of suitable treatment modalities. The restoration of structural and functional aspect of damaged neurological tissues at the lesion site in spinal cord has been challenging. Recent developments have showed tremendous potential of neural stem cell-based strategies to form a neuronal relay circuit across the injury gap which facilitates some levels of improvement in SCI condition. However, to provide better therapeutic responses, critical mass of grafted cells must survive for long-term and differentiate into neuronal cells with well-developed axonal networks. Hence, development of tissue specific biological neuronal constructs is highly desirable to provide mechanical and biological support for long-term survival and function of neurological cells within natural biological niche. In this study, we report development of a tissue specific neuronal constructs by culturing human neural precursor cells on decellularized meningeal scaffolds to provide suitable biological neuronal construct which can be used to support mechanical, structural and functional aspect of damaged spinal cord tissues. This particular tissue specific biological construct is immunologically tolerable and provides precisely orchestral three-dimensional platform to choreograph the long-distance axonal guidance and more organized neuronal cell growth. It passes sufficient mechanical and biological properties enriched with several crucial neurotrophins required for long-term survival and function of neurological cells which is required to form proper axonal bridge to regenerate the damaged axonal connectomes at lesion-site in SCI.

15.
World J Hepatol ; 10(1): 22-33, 2018 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-29399275

RESUMO

AIM: To develop appropriate humanized three-dimensional ex-vivo model system for drug testing. METHODS: Bioengineered humanized livers were developed in this study using human hepatic stem cells repopulation within the acellularized liver scaffolds which mimics with the natural organ anatomy and physiology. Six cytochrome P-450 probes were used to enable efficient identification of drug metabolism in bioengineered humanized livers. The drug metabolism study in bioengineered livers was evaluated to identify the absorption, distribution, metabolism, excretion and toxicity responses. RESULTS: The bioengineered humanized livers showed cellular and molecular characteristics of human livers. The bioengineered liver showed three-dimensional natural architecture with intact vasculature and extra-cellular matrix. Human hepatic cells were engrafted similar to the human liver. Drug metabolism studies provided a suitable platform alternative to available ex-vivo and in vivo models for identifying cellular and molecular dynamics of pharmacological drugs. CONCLUSION: The present study paves a way towards the development of suitable humanized preclinical model systems for pharmacological testing. This approach may reduce the cost and time duration of preclinical drug testing and further overcomes on the anatomical and physiological variations in xenogeneic systems.

16.
Tissue Cell ; 51: 14-23, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29622083

RESUMO

INTRODUCTION: Ethanol exposure to developing brain may alter the growth and differentiation of neurological cells resulting in unfavorable pathologies. Earlier studies have provided very limited mechanistic insights of cellular and molecular mechanisms which do not mimic with human situation due to varying cell types and poses potential challenges for investigation. Therefore, the present study was undertaken to evaluate the role of ABC transporters and heat shock proteins mediated response in human neural precursor cells (NPCs) and its lineages during proliferation and lineage differentiation against ethanol exposure. METHODS: Effect of ethanol exposure was examined for neuronal cell survival and variation in cellular phenotype during neurospheres development and lineage differentiation. Generation of reactive oxygen species, and variation in cell cycle was identified along with transcriptional profiling for pluripotent markers (Nestin, NCAM, Sox-2, and Notch-2), drug transporters (ABCB1 and ABCG2) and stress protein (HSP70) during ethanol exposure. RESULTS: ABC transporters as well as HSP70 mRNA expression was higher during proliferation as compared to differentiation with chronic ethanol (1 M) exposure (p < 0.01). Ethanol exposure resulted in higher variability in size and shape of developing neurospheres and decreased ability to form new neurosphere colonies. Significant changes were observed in dendrite development due to late ethanol exposure (p < 0.0001). CONCLUSION: The present study demonstrated significant role of ABC transporters and HSP70 proteins in providing defense against ethanol-induced damage in human neurological cells. However, the over-expression of ABC transporter and HSP-70 proteins during such pathological conditions do not provide complete defense and additional strategies are required to repair the damage.


Assuntos
Transportadores de Cassetes de Ligação de ATP/biossíntese , Diferenciação Celular/efeitos dos fármacos , Linhagem da Célula/efeitos dos fármacos , Depressores do Sistema Nervoso Central/toxicidade , Etanol/toxicidade , Proteínas de Choque Térmico HSP70/biossíntese , Células-Tronco Neurais/efeitos dos fármacos , Transportadores de Cassetes de Ligação de ATP/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Proteínas de Choque Térmico HSP70/efeitos dos fármacos , Humanos , Células-Tronco Neurais/metabolismo , Oxirredução/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos
17.
Tissue Cell ; 49(6): 638-647, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28947065

RESUMO

INTRODUCTION: Progress in understanding pathophysiological mechanisms and the development of targeted regenerative strategies have been hampered by the lack of predictive disease models, specifically for the conditions to which affected cell types are inaccessible. The present study has aimed to unearth the role of valproic acid (VPA) and mild hypothermia (MH) as promising strategy to enhance the neuroprotective mechanisms in undifferentiated and differentiated human neural precursor cells (hNPCs) against ethanol-induced damage. METHODS: 5mM VPA alone or in combination with MH (33°C) was used to prevent the damage in proliferating and differentiating hNPCs. CD133+ve enriched hNPCs were cultured in vitro and exposed to 1M chronic ethanol concentration for 72h and followed by VPA and MH treatment for 24h. Morphometric analysis was performed to identify changes in neurospheres development and neuronal cell phenotypes. Flow cytometry and RT-qPCR analysis was performed to investigate alterations in key molecular pathways involved in cell survival and signaling. RESULTS: Combination of VPA with MH displayed higher proportion of neuronal cell viability as compared to single treatment. Combination treatment was most effective in reducing apoptosis and reactive oxygen species levels in both the undifferentiated and differentiated hNPCs. VPA with MH significantly improved neuronal cell phenotype, active chromatin modeling, chaperon and multi-drug resistant pumps activity and expression of neuronal signaling molecules. CONCLUSION: The study provided an efficient and disease specific in vitro model and demonstrated that combined treatment with VPA and MH activates several neuroprotective mechanisms and provides enhanced protection against ethanol-induced damage in cultured undifferentiated and differentiated hNPCs.


Assuntos
Depressores do Sistema Nervoso Central/toxicidade , Etanol/toxicidade , Hipotermia Induzida , Células-Tronco Neurais/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Ácido Valproico/farmacologia , Apoptose/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Humanos
18.
Sci Rep ; 7(1): 8539, 2017 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-28819176

RESUMO

The present study identifies the potential of highly biocompatible SF-GNP nano-conjugate to enhance the chemotherapeutic response to combat drug resistance in cancer cells. We developed a stable colloidal suspension of sorafenib-gold nanoconjugate (SF-GNP) of <10 nm size in aqueous medium for reverting the cancer drug resistance in SF-resistant HepG2 cells in a 3D ex-vivo model system. In-vivo biocompatibility assay of SF-GNPs showed absence of systemic toxicological effects including hematological, biochemical and histological parameters. More importantly, the histopathological analysis of vital organs such as liver, brain, lung, kidney and heart showed very least or no sign of inflammation, cell infiltration, necrosis, tissue disorganization or fibrotic reactions after intra-peritoneal administration of SF-GNP nanoconjugates in animals. However, SF-GNP nanoconjugates significantly reduced (>80%) the percentage cell survival and the size and number of SF resistant solid tumor colonies of HepG2 cells in 3D model system. The exposure of SF-GNP nanoconjugate to SF resistant HepG2 cell colonies also provided evidence for anti-proliferative effect and reversal of drug resistance by elucidating the molecular regulatory mechanisms of extracellular matrix factor (CD147), tumor growth factor (TGF-ß), hepatoma upregulated protein (hURP) and drug transporter (ABCG-2).


Assuntos
Materiais Biocompatíveis/administração & dosagem , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Ouro/química , Nanopartículas Metálicas/administração & dosagem , Nanoconjugados/administração & dosagem , Sorafenibe/administração & dosagem , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Materiais Biocompatíveis/química , Sobrevivência Celular/efeitos dos fármacos , Células Hep G2 , Hepatoblastoma/patologia , Humanos , Neoplasias Hepáticas/patologia , Masculino , Nanopartículas Metálicas/química , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica de Transmissão , Nanoconjugados/química , Nanoconjugados/ultraestrutura , Tamanho da Partícula , Ratos Wistar , Sorafenibe/química
19.
Basic Clin Neurosci ; 8(6): 453-466, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29942429

RESUMO

INTRODUCTION: The present study aimed to explore protective mechanisms of hypothermia against mild cold and heat stress on highly proliferative homogeneous human Neural Precursor Cells (NPCs) derived from Subventricular Zone (SVZ) of human fetal brain. METHODS: CD133+ve enriched undifferentiated and differentiated human NPCs were exposed to heat stress at 42°C. Then, Western-blot quantification was performed using Hsp-70 (70 kilodalton heat shock proteins) recombinant protein. Finally, changes in pluripotency and Hsp-70 expression were measured using immunofluorescence staining and RT-qPCR (Quantitative reverse transcription PCR) analysis, respectively. RESULTS: Heat stress resulted in abnormal neurospheres development. The apoptosis rate was enhanced during long-term in vitro culture of neurospheres. Neurogenic differentiation reduced and showed aberrent phenotypes during heat stress. After hypothermia treatment significant improvement in neurospheres and neuronal cell morphology was observed. CONCLUSION: Mild-hypothermia treatment induces attenuated heat shock response against heat stress resulting in induced HSP-70 expression that significantly improves structure and function of both undifferentiated human NPCs and differentiated neurons.

20.
Inflammation ; 39(3): 1198-204, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27107596

RESUMO

Chronic obstructive pulmonary disease (COPD) is a heterogeneous collection of conditions characterized by irreversible expiratory airflow limitation. The disease is interspersed with exacerbations; periods of acute symptomatic, physiological, and functional deterioration. The present study was designed to investigate the role of X-ray cross-complementing group 1 (XRCC1) and apurinic/apyrimidinic endonuclease 1 (APE1) polymorphisms and the risk of COPD. Blood samples from 354 unrelated subject (age range 18-60 years; 156 with COPD, 198 healthy controls) were collected. Genomic DNA was isolated and genotyped for XRCC1 Arg399Gln and APE1 Asp148Glu using a confronting two pair primers polymerase chain reaction. GA genotype of XRCC1 gene was found to be predominant in the COPD group compared to controls with 1.86-fold increased risk for COPD (OR 1.86, 95 % CI 1.20-2.88, p = 0.0013). TG genotype of APE1 was found to be predominant in COPD group compared to controls with the difference being statistically significant (OR 1.68, 95 % CI 1.08-2.61, p = 0.0043). The GA haplotype was found to be predominant in COPD than controls with a 2.19-fold significant increase (OR 2.19, 95 % CI 1.46-3.28, p = 0.003). Polymorphism in XRCC1 and APE1 gene is associated with an increased risk of COPD.


Assuntos
DNA Liase (Sítios Apurínicos ou Apirimidínicos)/genética , Proteínas de Ligação a DNA/genética , Polimorfismo Genético , Doença Pulmonar Obstrutiva Crônica/genética , Adolescente , Adulto , Estudos de Casos e Controles , Predisposição Genética para Doença , Genoma Humano/genética , Genótipo , Humanos , Pessoa de Meia-Idade , Proteína 1 Complementadora Cruzada de Reparo de Raio-X
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