In vitro effects of mechanical stimulation and photobiomodulation on osteoblastic cell function: A proof of concept study

2017 ◽  
Vol 27 (1) ◽  
pp. 29-41 ◽  
Author(s):  
Rochaya Chintavalakorn ◽  
Anak Khantachawana ◽  
Kwanchanok Viravaidya-Pasuwat ◽  
Peerapong Santiwong ◽  
Rudee Surarit
Keyword(s):  
Mechanical Stimulation ◽  
Cell Function ◽  
Osteoblastic Cell ◽  
Proof Of Concept ◽  
Concept Study ◽  
In Vitro Effects

scholarly journals Microvalve bioprinting of MSC-Chondrocyte Co-Cultures

2021 ◽  
Author(s):  
Joseph Dudman ◽  
Ana Marina Ferreira ◽  
Piergiorgio Gentile ◽  
Xiao Wang ◽  
Kenneth Dalgarno
Keyword(s):  
High Throughput Screening ◽  
Cellular Therapy ◽  
Cell Types ◽  
Inhibitory Effect ◽  
Proof Of Concept ◽  
Concept Study ◽  
Indirect Immunofluorescence Staining ◽  
Chondrocyte Implantation ◽  
Tissue Models

Recent improvements within the fields of high-throughput screening and 3D tissue culture have provided the possibility of developing in vitro micro-tissue models that can be used to study diseases and screen potential new therapies. This paper reports a proof of concept study on the use of microvalve-based bioprinting to create laminar MSC-chondrocyte co-cultures as an in vitro model of autologous chondrocyte implantation (ACI), an established cellular therapy for osteoarthritis. Microvalve-based bioprinting uses microvalves to deposit cells suspended in a liquid in a consistent and repeatable manner. In this case MSCs and chondrocytes have been sequentially deposited into an insert based transwell system in order to create a laminar co-culture, with variations in the ratios of the cell types used to investigate the potential for MSCs to stimulate improved repair. Histological and indirect immunofluorescence staining revealed the formation of dense tissue structures within the chondrocyte and MSC-chondrocyte cell co-cultures, alongside the establishment of a proliferative region at the base of the tissue. No stimulatory or inhibitory effect in terms of ECM production was observed through the introduction of MSCs, although the potential for an immunomodulatory benefit remains. This proof-of-concept study therefore provides a novel method to enable the scalable production of therapeutically relevant micro-tissue models that can be used for in vitro research to optimise ACI procedures.

scholarly journals Raman spectroscopy for the preoperative diagnosis of thyroid cancer and its subtypes: An in vitro proof-of-concept study

Cytopathology ◽  
2018 ◽  
Vol 30 (1) ◽  
pp. 51-60 ◽  
Author(s):  
Declan O'Dea ◽  
Massimo Bongiovanni ◽  
Gerasimos P. Sykiotis ◽  
Panos G. Ziros ◽  
Aidan D. Meade ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Thyroid Cancer ◽  
Preoperative Diagnosis ◽  
Proof Of Concept ◽  
Concept Study ◽  
Diagnosis Of Thyroid Cancer

scholarly journals Role of UDP-Sugar Receptor P2Y14 in Murine Osteoblasts

2020 ◽  
Vol 21 (8) ◽  
pp. 2747 ◽  
Author(s):  
Nicholas Mikolajewicz ◽  
Svetlana V. Komarova
Keyword(s):  
Mechanical Stimulation ◽  
Calcium Signalling ◽  
Free Calcium ◽  
Osteoblastic Cell ◽  
Uridine Diphosphate ◽  
P2 Receptor ◽  
Mineral Density ◽  
Osteoblastic Cell Line ◽  
Camp Levels

The purinergic (P2) receptor P2Y14 is the only P2 receptor that is stimulated by uridine diphosphate (UDP)-sugars and its role in bone formation is unknown. We confirmed P2Y14 expression in primary murine osteoblasts (CB-Ob) and the C2C12-BMP2 osteoblastic cell line (C2-Ob). UDP-glucose (UDPG) had undiscernible effects on cAMP levels, however, induced dose-dependent elevations in the cytosolic free calcium concentration ([Ca2+]i) in CB-Ob, but not C2-Ob cells. To antagonize the P2Y14 function, we used the P2Y14 inhibitor PPTN or generated CRISPR-Cas9-mediated P2Y14 knockout C2-Ob clones (Y14KO). P2Y14 inhibition facilitated calcium signalling and altered basal cAMP levels in both models of osteoblasts. Importantly, P2Y14 inhibition augmented Ca2+ signalling in response to ATP, ADP and mechanical stimulation. P2Y14 knockout or inhibition reduced osteoblast proliferation and decreased ERK1/2 phosphorylation and increased AMPKα phosphorylation. During in vitro osteogenic differentiation, P2Y14 inhibition modulated the timing of osteogenic gene expression, collagen deposition, and mineralization, but did not significantly affect differentiation status by day 28. Of interest, while P2ry14-/- mice from the International Mouse Phenotyping Consortium were similar to wild-type controls in bone mineral density, their tibia length was significantly increased. We conclude that P2Y14 in osteoblasts reduces cell responsiveness to mechanical stimulation and mechanotransductive signalling and modulates osteoblast differentiation.

scholarly journals In vitro effects of combined iron chelation, antibiotics and matrix disruption on clinical isolates of Pseudomonas aeruginosa

2019 ◽  
Vol 75 (3) ◽  
pp. 586-592
Author(s):  
Arun Nair ◽  
Audrey Perry ◽  
John D Perry ◽  
F Kate Gould ◽  
Julie Samuel
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Iron Chelation ◽  
Proof Of Concept ◽  
Crystal Violet Assay ◽  
Clear Advantage ◽  
Biofilm Disruption ◽  
In Vitro Effects

Abstract Objectives Pseudomonas aeruginosa is an important pathogen in chronic suppurative respiratory diseases, with adverse effects on severity, healthcare utilization and quality of life. Aerosolized combined biofilm disruption and iron chelators offer novel proof-of-concept for improving airway antimicrobial efficacy. Our aim was to assess the activity of desferrioxamine, Dornase alfa (DNase) and antibiotics on biofilm formation and against mature preformed biofilms of P. aeruginosa. Methods Fifty-six isolates of P. aeruginosa were screened for biofilm production and seven isolates with varying capacity to form biofilms were referred for further study. Three antibiotics (colistin, tobramycin and ciprofloxacin) as well as desferrioxamine and DNase were assessed for their ability to prevent biofilm formation using the crystal violet assay. The same method was used to assess their impact on mature biofilms. Each agent, as well as combinations of these agents, was also assessed for its effect on the metabolic activity and viability of preformed P. aeruginosa biofilm by the resazurin reduction assay and by performing viable counts. Results Antibiotics alone prevented the development of biofilms and partly reduced the viability of mature biofilms. Desferrioxamine and DNase did not reduce biofilm formation. For most isolates, desferrioxamine and DNase did not offer any clear advantage over the use of antibiotics alone with respect to reducing the viability of Pseudomonas biofilms. Conclusions Colistin, tobramycin and ciprofloxacin prevented biofilm formation by P. aeruginosa and reduced the viability of mature biofilms. For most isolates, there was no clear advantage of combining these antimicrobials with desferrioxamine or DNase.

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