saliva-coated hydroxyapatite (sha) disks  (Clarkson Chromatography Products)

Journal: Arthritis Research & Therapy

Article Title: Pro-inflammatory effects of human apatite crystals extracted from patients suffering from calcific tendinopathy

doi: 10.1186/s13075-021-02516-9

Figure Lengend Snippet: a Representative images of calcific powders obtained by scanning electron microscopy. (A1) Human sample collected after a percutaneous needle lavage under ultrasound. (A2, A3) Synthetic hydroxyapatite before and after grinding, respectively. b IL-1β release after 6h stimulation of CD14+ monocytes, MCSF-macrophages, GM-CSF/IFNγ-macrophages, and tenocytes by crystals (1mg/ml) and parallel cell death rate. sHA, synthetic hydroxyapatite; CAL, human calcification; CT, control group without crystal; LPS, lipopolysaccharide; ND, not detectable. n = 3. * p ≤ 0.05

Article Snippet: Synthetic hydroxyapatite crystals (sHA) (REF-289396, Millipore Sigma, Switzerland) were first crushed using a mortar and pestle to obtain a powder close to the human crystals.

Techniques: Electron Microscopy, Control

Journal: Arthritis Research & Therapy

Article Title: Pro-inflammatory effects of human apatite crystals extracted from patients suffering from calcific tendinopathy

doi: 10.1186/s13075-021-02516-9

Figure Lengend Snippet: IL-1β production by THP-1 cells after crystal stimulation. a Release of IL-1β by THP-1 cells cultured during 6 h with crystals (1mg/ml) in media with or without 10% fetal bovine serum (FBS) ( n = 3). b Kinetic of IL-1β production by THP-1 cells cultured with a patient’s calcification (CAL, 1mg/ml) and synthetic hydroxyapatite (sHA, 1 mg/ml) or without crystal (CT) ( n = 3). c IL-1β release in cultures containing only crystals compared to THP-1 cells after crystal stimulation ( N = 3 patients). d IL-1β quantification in supernatants of THP-1 after 6 h of crystal stimulation with increasing concentrations of calcifications of 9 different patients ( N = 9). sHA, synthetic hydroxyapatite; CAL, human calcification; CT, control condition without crystals. * p ≤ 0.05

Article Snippet: Synthetic hydroxyapatite crystals (sHA) (REF-289396, Millipore Sigma, Switzerland) were first crushed using a mortar and pestle to obtain a powder close to the human crystals.

Techniques: Cell Culture, Control

Journal: Arthritis Research & Therapy

Article Title: Pro-inflammatory effects of human apatite crystals extracted from patients suffering from calcific tendinopathy

doi: 10.1186/s13075-021-02516-9

Figure Lengend Snippet: THP-1 gene expression after 6 h of stimulation by crystals (synthetic hydroxyapatite (sHA) or human calcification (CAL) at 1 mg/mL). Gene expression is expressed in relative expression (2 −ΔCt ) with the HPRT gene used as a reference. CT, control condition without crystal. * p ≤ 0.05 ( n = 3)

Article Snippet: Synthetic hydroxyapatite crystals (sHA) (REF-289396, Millipore Sigma, Switzerland) were first crushed using a mortar and pestle to obtain a powder close to the human crystals.

Techniques: Gene Expression, Expressing, Control

Journal: Arthritis Research & Therapy

Article Title: Pro-inflammatory effects of human apatite crystals extracted from patients suffering from calcific tendinopathy

doi: 10.1186/s13075-021-02516-9

Figure Lengend Snippet: Effects of a NF-κB inhibitor and a caspase-1 inhibitor on the secretion of IL-1β by THP-1 macrophages. a Quantification of IL-1β released into the medium after 6h of stimulation by crystals (1mg/ml) in the presence or absence of a NF-κB inhibitor (BAY-11-7085, 10μM) or a caspase-1 inhibitor (Z-YVAD-FMK, 10 μM) ( n = 3). b , c Effects of crystals and inhibitors on NLRP3 and IL-1β assessed by Western Blot on cell lysates and supernatants (SN). One representative patient out of 3 is presented in b , and quantification of cleaved-IL-1β in cell lysates and supernatant is presented in c ( N = 3 patients), with GAPDH used as an invariant protein. iNF-κB, NF-κB inhibitor; iCasp1, caspase-1 inhibitor; inh, inhibitor; sHA, synthetic hydroxyapatite; CAL, human calcification; CT, control condition without crystals. * p ≤ 0.05

Article Snippet: Synthetic hydroxyapatite crystals (sHA) (REF-289396, Millipore Sigma, Switzerland) were first crushed using a mortar and pestle to obtain a powder close to the human crystals.

Techniques: Western Blot, Control