Integration of a miniaturized DMMB assay with high-throughput screening for identifying regulators of proteoglycan metabolism

Sun, Yi; Tsui, Yuen-Kee; Yu, Mengqi; Lyu, Minmin; Cheung, Kenneth M.C.; Kao, Richard Yi Tsun; Leung, Victor

doi:10.1038/s41598-022-04805-y

Cited by 2 publications

(1 citation statement)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…After pipetting, the absorbance was measured at 656 nm using a microplate reader. 29 For ultrastructure assessment lyophilized and intact samples were prepared for scanning electron microscopy (SEM). The intact samples were fixed using 2.5% glutaraldehyde (Sigma-Aldrich, USA) in 0.1 M PBS and 4% formaldehyde at 4 °C overnight, they were gradually dehydrated via an increasing graded series of ethanol (50,70,90 and 100%) and kept overnight to be air-dried in a fume hood.…”

Section: Characterization Of Decellularized Cartilagementioning

confidence: 99%

An update on a technical method of cartilage decellularization: (a physical-based protocol)

Dortaj,

Vaez,

Hassanpour-Dehnavie

et al. 2024

Bioimpacts

View full text Add to dashboard Cite

Introduction: Despite advances in orthopedic surgery, the lack of effective conventional treatment for cartilage defects has led to research in cartilage tissue engineering. One of the interesting topics is the use of decellularized extracellular matrix (ECM) as a suitable natural scaffold that supports the growth and function of cells cultured in it. A concern with decellularization protocols, especially those with high detergent concentrations, is the disruption of native ECM, which has deleterious effects on subsequent scaffold recellularization. Therefore, this study focused on optimizing cartilage decellularization by physical methods without the use of ionic detergents. Methods: The bovine tracheal cartilage fragments were decellularized by a combination of 8 cycles of freeze-thaw and ultrasound techniques. Then, the tissues were immersed and shaken in 0.25% trypsin for 24 hours. Efficient cell removal and preservation of ECM were confirmed by histological and cytocompatibility assessments. The in-vivo studies were performed to evaluate the biocompatibility and bioactivity of the scaffold. Results: The histological assessments indicated the appropriate cytocompatibility and the fibroblast cell culture study demonstrated that cells were able to proliferate and migrate on the decellularized cartilage. In-vivo evaluation also showed a reduced adverse immune response, including leukocyte infiltration into the ECM. Conclusions: These results suggest that a cartilage scaffold created using a physical decellularization protocol that efficiently removes cells while preserving the native ECM can be a suitable scaffold for cartilage reconstruction. The main advantage of this protocol is the absence of potentially toxic chemicals in the tissues.

show abstract

Section: Characterization Of Decellularized Cartilagementioning

confidence: 99%