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1.
Gene ; 788: 145662, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33887373

RESUMO

INTRODUCTION: Culture conditions and differentiation cocktails may facilitate cell maturation and extracellular matrix (ECM) secretion and support the production of engineered fibroblastic tissues with applications in ligament regeneration. The objective of this study is to investigate the potential of two connective tissue-related ligands (i.e., BMP6 and GDF5) to mediate collagenous ECM synthesis and tissue maturation in vitro under normoxic and hypoxic conditions based on the hypothesis that BMP6 and GDF5 are components of normal paracrine signalling events that support connective tissue homeostasis. METHODS: Human adipose-derived MSCs were seeded on 3D-printed medical-grade polycaprolactone (PCL) scaffolds using a bioreactor and incubated in media containing GDF5 and/or BMP6 for 21 days in either normoxic (5% oxygen) or hypoxic (2% oxygen) conditions. Constructs were harvested on Day 3 and 21 for cell viability analysis by live/dead staining, structural analysis by scanning electron microscopy, mRNA levels by RTqPCR analysis, and in situ deposition of proteins by immunofluorescence microscopy. RESULTS: Pro-fibroblastic gene expression is enhanced by hypoxic culture conditions compared to normoxic conditions. Hypoxia renders cells more responsive to treatment with BMP6 as reflected by increased expression of ECM mRNA levels on Day 3 with sustained expression until Day 21. GDF5 was not particularly effective either in the absence or presence of BMP6. CONCLUSIONS: Fibroblastic differentiation of MSCs is selectively enhanced by BMP6 and not GDF5. Environmental factors (i.e., hypoxia) also influenced the responsiveness of cells to this morphogen.


Assuntos
Proteína Morfogenética Óssea 6/farmacologia , Técnicas de Cultura de Células/métodos , Fibroblastos/citologia , Fator 5 de Diferenciação de Crescimento/farmacologia , Células-Tronco Mesenquimais/citologia , Reatores Biológicos , Diferenciação Celular/efeitos dos fármacos , Hipóxia Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Fibroblastos/química , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/química , Células-Tronco Mesenquimais/efeitos dos fármacos , Alicerces Teciduais
2.
Nanomaterials (Basel) ; 10(11)2020 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-33203046

RESUMO

Bacterial biofilms are involved in most device-associated infections and remain a challenge for modern medicine. One major approach to addressing this problem is to prevent the formation of biofilms using novel antimicrobial materials, device surface modification or local drug delivery; however, successful preventive measures are still extremely limited. The other approach is concerned with treating biofilms that have already formed on the devices; this approach is the focus of our manuscript. Treating biofilms associated with medical devices has unique challenges due to the biofilm's extracellular polymer substance (EPS) and the biofilm bacteria's resistance to most conventional antimicrobial agents. The treatment is further complicated by the fact that the treatment must be suitable for applying on devices surrounded by host tissue in many cases. Nanomaterials have been extensively investigated for preventing biofilm formation on medical devices, yet their applications in treating bacterial biofilm remains to be further investigated due to the fact that treating the biofilm bacteria and destroying the EPS are much more challenging than preventing adhesion of planktonic bacteria or inhibiting their surface colonization. In this highly focused review, we examined only studies that demonstrated successful EPS destruction and biofilm bacteria killing and provided in-depth description of the nanomaterials and the biofilm eradication efficacy, followed by discussion of key issues in this topic and suggestion for future development.

3.
Nanomaterials (Basel) ; 10(5)2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32429310

RESUMO

Multifunctional scaffolds are becoming increasingly important in the field of tissue engineering. In this research, a composite material is developed using polycaprolactone (PCL) and detonation nanodiamond (ND) to take advantage of the unique properties of ND and the biodegradability of PCL polymer. Different ND loading concentrations are investigated, and the physicochemical properties of the composites are characterized. ND-PCL composite films show a higher surface roughness and hydrophilicity than PCL alone, with a slight decrease in tensile strength and a significant increase in degradation. Higher loading of ND also shows a higher osteoblast adhesion than the PCL alone sample. Finally, we show that the ND-PCL composites are successfully extruded to create a 3D scaffold demonstrating their potential as a composite material for tissue regeneration.

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