Journal: JOR Spine

Article Title: In silico modeling the potential clinical effect of growth factor treatment on the metabolism of human nucleus pulposus cells

doi: 10.1002/jsp2.1352

Figure Lengend Snippet: (A) Human nucleus pulposus (NP) cells were isolated from four patients undergoing discectomy procedures. Following monolayer expansion, cell spheroids were formed in 96‐well plates coated with a thin layer of 2% agarose to prevent attachment. (B) Cell spheroids were cultured in standard expansion media (XPAN) for 1 week prior to 2 weeks under growth differentiation factor‐5 (GDF‐5) stimulation. (C) After GDF‐5 stimulation, individual spheroids underwent metabolic analysis using a Seahorse XFe96 analyzer and biochemical analysis. (D) Metabolic rates for each experimental group were then computed in silico to predict the donor‐specific reparative effects of GDF‐5 on extracellular matrix and subsequent impact on the nutrient microenvironment.

Article Snippet: Experimental groups consisted of a XPAN‐only (LG‐DMEM supplemented with 10% FBS and 2% Pen‐Strep) control (ctr) and the three clinically investigated GDF‐5 (PeproTech, Thermo Fisher Scientific) doses, consisting of XPAN supplemented with 0.25, 1, and 2 mg. Doses were normalized to the average cell population of the human NP to yield a working concentration of 0.75, 3, and 6 μg per spheroid for the 0.25, 1, and 2 mg groups, respectively.

Techniques: Isolation, Cell Culture, In Silico

Journal: JOR Spine

Article Title: In silico modeling the potential clinical effect of growth factor treatment on the metabolism of human nucleus pulposus cells

doi: 10.1002/jsp2.1352

Figure Lengend Snippet: (A) Spheroid viability was assessed across the experimental groups using live/dead staining to ensure viability remained high in each group prior to performing metabolic rate measurements. (B) Oxygen consumption rates (OCR, nmol/million cells/h) and (C) lactate production rates (LPR, nmol/million cells/h) for NP cell spheroids in either XPAN media as an untreated control (ctr) or stimulated with three different concentrations of growth differentiation factor‐5 (GDF‐5) (0.25, 1, and 2 mg) ( N = 4). While there is an apparent trend of increasing OCR and LPR with GDF‐5 concentration, only the 2 mg group had a significantly higher OCR compared to the ctr ( p = 0.045).

Article Snippet: Experimental groups consisted of a XPAN‐only (LG‐DMEM supplemented with 10% FBS and 2% Pen‐Strep) control (ctr) and the three clinically investigated GDF‐5 (PeproTech, Thermo Fisher Scientific) doses, consisting of XPAN supplemented with 0.25, 1, and 2 mg. Doses were normalized to the average cell population of the human NP to yield a working concentration of 0.75, 3, and 6 μg per spheroid for the 0.25, 1, and 2 mg groups, respectively.

Techniques: Staining, Control, Concentration Assay

Journal: JOR Spine

Article Title: In silico modeling the potential clinical effect of growth factor treatment on the metabolism of human nucleus pulposus cells

doi: 10.1002/jsp2.1352

Figure Lengend Snippet: (A) Glycosaminoglycan (GAG) production rates and (B) collagen production rates for cell spheroids in either XPAN media (control [ctr]) or stimulated with three different concentrations of growth differentiation factor‐5 (GDF‐5) (0.25, 1, and 2 mg) ( N = 4). While no statistically significant difference was detected between experimental groups, donor‐specific colors are used to highlight donor variability and trends in donor‐specific response across both GAG and collagen. (C) Corresponding histological evaluation for Donor 2 using hematoxylin and eosin (H&E) to stain for cells, alcian blue (AB) to stain for GAG and picrosirius red (PSR) to stain for collagen. Scale bar is 200 μm.

Article Snippet: Experimental groups consisted of a XPAN‐only (LG‐DMEM supplemented with 10% FBS and 2% Pen‐Strep) control (ctr) and the three clinically investigated GDF‐5 (PeproTech, Thermo Fisher Scientific) doses, consisting of XPAN supplemented with 0.25, 1, and 2 mg. Doses were normalized to the average cell population of the human NP to yield a working concentration of 0.75, 3, and 6 μg per spheroid for the 0.25, 1, and 2 mg groups, respectively.

Techniques: Control, Staining

Journal: iScience

Article Title: Time- and cell-specific activation of BMP signaling restrains chondrocyte hypertrophy

doi: 10.1016/j.isci.2024.110537

Figure Lengend Snippet:

Article Snippet: Recombinant GDF-5 , Peprotech , Cat. No. 120-01.

Techniques: Recombinant, Plasmid Preparation, RNA Sequencing Assay, Software