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1.
Mol Biol Rep ; 51(1): 570, 2024 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-38658405

RESUMO

INTRODUCTION: Spinal cord injury (SCI) leads to significant destruction of nerve tissue, causing the degeneration of axons and the formation of cystic cavities. This study aimed to examine the characteristics of human umbilical cord-derived mesenchymal stem cells (HUCMSCs) cultured in a serum-free conditioned medium (CM) and assess their effectiveness in a well-established hemitransection SCI model. MATERIALS AND METHODS: In this study, HUCMSCs cultured medium was collected and characterized by measuring IL-10 and identifying proteomics using mass spectroscopy. This collected serum-free CM was further used in the experiments to culture and characterize the HUMSCs. Later, neuronal cells derived from CM-enriched HUCMSC were tested sequentially using an injectable caffeic acid-bioconjugated gelatin (CBG), which was further transplanted in a hemitransection SCI model. In vitro, characterization of CM-enriched HUCMSCs and differentiated neuronal cells was performed using flow cytometry, immunofluorescence, electron microscopy, and post-transplant analysis using immunohistology analysis, qPCR, in vivo bioluminescence imaging, and behavioral analysis using an infrared actimeter. RESULTS: The cells that were cultured in the conditioned media produced a pro-inflammatory cytokine called IL-10. Upon examining the secretome of the conditioned media, the Kruppel-like family of KRAB and zinc-finger proteins (C2H2 and C4) were found to be activated. Transcriptome analysis also revealed an increased expression of ELK-1, HOXD8, OTX2, YY1, STAT1, ETV7, and PATZ1 in the conditioned media. Furthermore, the expression of Human Stem-101 confirmed proliferation during the first 3 weeks after transplantation, along with the migration of CBG-UCNSC cells within the transplanted area. The gene analysis showed increased expression of Nestin, NeuN, Calb-2, Msi1, and Msi2. The group that received CBG-UCNSC therapy showed a smooth recovery by the end of week 2, with most rats regaining their walking abilities similar to those before the spinal cord injury by week 5. CONCLUSIONS: In conclusion, the CBG-UCNSC method effectively preserved the integrity of the transplanted neuronal-like cells and improved locomotor function. Thus, CM-enriched cells can potentially reduce biosafety risks associated with animal content, making them a promising option for clinical applications in treating spinal cord injuries.


Assuntos
Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Traumatismos da Medula Espinal , Transcriptoma , Cordão Umbilical , Traumatismos da Medula Espinal/terapia , Traumatismos da Medula Espinal/metabolismo , Traumatismos da Medula Espinal/genética , Células-Tronco Mesenquimais/metabolismo , Meios de Cultivo Condicionados/farmacologia , Cordão Umbilical/citologia , Cordão Umbilical/metabolismo , Humanos , Animais , Transplante de Células-Tronco Mesenquimais/métodos , Transcriptoma/genética , Ratos , Secretoma/metabolismo , Diferenciação Celular , Neurônios/metabolismo , Modelos Animais de Doenças , Interleucina-10/genética , Interleucina-10/metabolismo , Células Cultivadas , Proteômica/métodos
2.
Braz J Microbiol ; 42(3): 825-34, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24031698

RESUMO

A new Streptomyces sp. IF 5 was isolated from the feather dumped soil and found to have a tremendous keratinase activity. The strain enabled the degradation of the chicken feathers very effectively in 60 h. The 16S rRNA sequence of 1474 bp long was submitted to the National centre for Biotechnological information. The keratinolytic activity in the culture medium was 1181 U/ml. The release and analyses of sulphydryl groups in the culture medium evident the degradation activity by the Streptomyces sp. IF 5. The idea of the present study was to use the degraded chicken feathers as the substrate for the growth and cultivation of microorganisms. We have designed a very economical culture medium that includes the usage of some basal salts alone and degraded chicken feathers (10 g/l). The results of the specific growth rate of the tested microbes confirm the usage of the new designed medium for microbial culturing.

3.
Exp Mol Med ; 48: e209, 2016 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-26869025

RESUMO

The success of regeneration attempt is based on an ideal combination of stem cells, scaffolding and growth factors. Tissue constructs help to maintain stem cells in a required area for a desired time. There is a need for easily obtainable cells, potentially autologous stem cells and a biologically acceptable scaffold for use in humans in different difficult situations. This study aims to address these issues utilizing a unique combination of stem cells from gingiva and a hydrogel scaffold, based on a natural product for regenerative application. Human gingival mesenchymal stem cells (HGMSCs) were, with due induction, differentiated to neuronal lineages to overcome the problems associated with birth tissue-related stem cells. The differentiation potential of neuronal lineages was confirmed with suitable specific markers. The properties of mesenchymal stem cells in encapsulated form were observed to be similar to free cells. The encapsulated cells (3D) were then subjected to differentiation into neuronal lineages with suitable inducers, and the morphology and gene expression of transient cells were analyzed. HGMSCs was differentiated into neuronal lineages as both free and encapsulated forms without any significant differences. The presence of Nissl bodies and the neurite outgrowth confirm the differentiation. The advantages of this new combination appear to make it a promising tissue construct for translational application.


Assuntos
Diferenciação Celular , Gengiva/citologia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Neurônios/citologia , Regeneração , Adulto , Biomarcadores , Técnicas de Cultura de Células , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato , Células-Tronco Mesenquimais/metabolismo , Fenótipo , Engenharia Tecidual , Alicerces Teciduais , Adulto Jovem
4.
Braz. j. microbiol ; 42(3): 825-834, July-Sept. 2011. ilus, tab
Artigo em Inglês | LILACS | ID: lil-607511

RESUMO

A new Streptomyces sp. IF 5 was isolated from the feather dumped soil and found to have a tremendous keratinase activity. The strain enabled the degradation of the chicken feathers very effectively in 60 h. The 16S rRNA sequence of 1474 bp long was submitted to the National centre for Biotechnological information. The keratinolytic activity in the culture medium was 1181 U/ml. The release and analyses of sulphydryl groups in the culture medium evident the degradation activity by the Streptomyces sp. IF 5. The idea of the present study was to use the degraded chicken feathers as the substrate for the growth and cultivation of microorganisms. We have designed a very economical culture medium that includes the usage of some basal salts alone and degraded chicken feathers (10 g/l). The results of the specific growth rate of the tested microbes confirm the usage of the new designed medium for microbial culturing.


Assuntos
Animais , Sequência de Bases , Ensaios Enzimáticos Clínicos , Microbiologia Ambiental , Análise de Alimentos , Microbiologia de Alimentos , Genética Microbiana , Meios de Cultura/isolamento & purificação , Estabilidade de RNA , Streptomyces/isolamento & purificação , Galinhas , Ativação Enzimática , Alimentos , Amostras de Alimentos , Métodos , Métodos
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