The control of cell behaviour in an effort to create highly homogeneous cultures is becoming an area of intense research, both to elucidate fundamental biology and for regenerative applications. The extracellular matrix (ECM) controls many cellular processes in vivo, and as such is a rich source of cues that may be translated in vitro. Herein, we describe the creation of cell culture coatings from porcine decellularised hyaline cartilage through enzymatic digestion. Surprisingly, heat-mediated sterilisation created a coating with the capacity to rapidly and robustly induce chondrogenic differentiation of human periosteal cells. This differentiation was validated through the alteration of cell phenotype from a fibroblastic to a cuboidal/cobblestone chondrocyte-like appearance. Moreover, chondrogenic gene expression further supported this observation, where cells cultured on heat sterilised ECM-coated plastic displayed higher expression of COL2A1, ACAN and PRG4 (p < 0.05) compared to non-coated plastic cultures. Interestingly, COL2A1 and ACAN expression in this context were sensitive to initial cell density; however, SOX9 expression appeared to be mainly driven by the coating independent of seeding density. The creation of a highly chondrogenic coating may provide a cost-effective solution for the differentiation and/or expansion of human chondrocytes aimed towards cartilage repair strategies.
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http://dx.doi.org/10.3390/bioengineering9050203 | DOI Listing |
J Vis Exp
January 2025
Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School;
A method to quantitate the stabilization of Mitochondria-Associated endoplasmic reticulum Membranes (MAMs) in a 3-dimensional (3D) neural model of Alzheimer's disease (AD) is presented here. To begin, fresh human neuro progenitor ReN cells expressing β-amyloid precursor protein (APP) containing familial Alzheimer's disease (FAD) or naïve ReN cells are grown in thin (1:100) Matrigel-coated tissue culture plates. After the cells reach confluency, these are electroporated with expression plasmids encoding red fluorescence protein (RFP)-conjugated mitochondria-binding sequence of AKAP1(34-63) (Mito-RFP) that detects mitochondria or constitutive MAM stabilizers MAM 1X or MAM 9X that stabilize tight (6 nm ± 1 nm gap width) or loose (24 nm ± 3 nm gap width) MAMs, respectively.
View Article and Find Full Text PDFUltrason Sonochem
January 2025
Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy; Polito(BIO)Med Lab, Politecnico di Torino, 10129 Turin, Italy. Electronic address:
In the context of molecular imaging, the present work explores an innovative platform made of lipid-coated nanocrystals as contrast-enhanced agent for both ultrasound imaging and sonoluminescence. At first, the dynamics of gas bubbles generation and cavitation under insonation with either pristine or lipid-coated nanocrystals (ZnO-Lip) are described, and the differences between the two colloidal systems are highlighted. These ZnO-Lip show an unprecedented ability to assist cavitation, which is reflected in enhanced sonoluminescent light emission with respect to the pristine nanocrystals or the pure water.
View Article and Find Full Text PDFAnn Biomed Eng
January 2025
Department of Biomedical Engineering, Yildiz Technical University, Esenler, 34220, Istanbul, Türkiye.
Titanium (Ti)-based materials are favored for hard tissue applications, yet their bioinertness limits their success. This study hypothesizes that functionalizing Ti materials with chitosan nano/microspheres and calcitriol (VD) will enhance their bioactivity by improving cellular activities and mineralization. To test this, chitosan particles were applied uniformly onto Ti surfaces using electrophoretic deposition (EPD) at 20 V for 3 minutes.
View Article and Find Full Text PDFEur J Trauma Emerg Surg
January 2025
Dr C Lal Hospital, Ambala Cantt, Haryana, 133001, India.
Purpose: Tibial open shaft fractures are very common and susceptible to infection, which can lead to significant morbidity especially infection and non-union. Antibiotic coated nail is one option for fixing open shaft tibial fractures to minimise infection. This study aimed to compare the clinical outcome of Gentamicin-coated tibial nails versus regular unreamed interlocking tibial nails in the treatment of type I and II tibial open fractures.
View Article and Find Full Text PDFDrug Test Anal
January 2025
European Monitoring Center for Emerging Doping Agents, German Sport University Cologne, Cologne, Germany.
A cost minimized immunoaffinity protocol was developed, which allows the direct purification of ERAs (urinary and recombinant human EPO, Darbepoetin, EPO-Fc, CERA) from human urine. The method applies magnetic beads and needs no covalent immobilization of the capture antibody. It requires only 10 mL of urine, 1 μg of anti-EPO antibody, and 25 μL of bead slurry.
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