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MP58-09 HYDROXYAPATITE INDUCES CALCIUM OXALATE TOLERANCE IN PRIMARY HUMAN MONOCYTES
THE JOURNAL OF UROLOGYâ
Vol. 195, No. 4S, Supplement, Monday, May 9, 2016
mineral formations appreciated in bone, cementum and dentin. Multiscale analyses of the spatial distribution of organic and inorganic components of biominerals across organ systems may provide insights into the pathogenesis of nephrolithiasis.
e779
partially explain the lack of papillary inflammation in the pathogenesis of Randall’s plaque.
Source of Funding: NIH NIDDK 1P20DK100863 (MLS); NIH NIDCR R01DE022032 (SPH); NIH/NCRR S10RR026645 (SPH)
MP58-09 HYDROXYAPATITE INDUCES CALCIUM OXALATE TOLERANCE IN PRIMARY HUMAN MONOCYTES Paul Dominguez Gutierrez, Sergei Kusmartsev, Benjamin Canales, Vincent Bird, Johannes Vieweg, Saeed Khan*, Gainesville, FL INTRODUCTION AND OBJECTIVES: Although most crystal deposits within tissue produce inflammation, renal interstitial hydroxyapatite deposits most like do not, accumulating as Randall’s plaque. To further explore this lack of response, we investigated the effect of oxalate, hydroxyapatite (HA), and combination of both on time dependent, immunological responses in human monocytes, the precursors to tissue macrophages. METHODS: Primary human monocytes and human monocytic cell line THP-1 cells were exposed to varying concentrations of soluble potassium oxalate (KOx) or insoluble CaOx and HA crystals. Primary human monocytes cells were pre-treated with 100 ug/ml of HA or CaOx followed by secondary treatment with 100 ug/ml HA, 100 ug/ml CaOx, and 1 ug/ml LPS. Cells were collected at various times after various treatments, and RNA was analyzed by quantitative real time PCR. Furthermore, monocytes function to enhance macrophage function and differentiation in the presence of CaOx where observed. RESULTS: Primary monocytes and THP-1 cells responded strongly to CaOx in a dose dependent manner producing TNFa, IL-1b, IL-8, and IL-10 with little to no response to KOx and HA. Pre-exposure to HA had little effect on human monocytes response to CaOx and LPS exposure; however, pre-exposure to CaOx followed by HA negated cytokine production. CONCLUSIONS: CaOx stimulates monocytes to produce cytokines and chemokines which function to stimulate and enhance macrophage uptake of CaOx; furthermore, monocytes exposed to CaOx go on to differentiate into macrophages. Furthermore, CaOx recognition by monocytes appears to be specific. Monocytes exposed to CaOx followed by LPS produce more cytokiens than LPS; however, monocytes exposed to LPS followed CaOx display a greatly downregulate response compared to LPS alone. HA, a component of Randal’s plaques, does not stimulate monocytes; however, it functions to specifically downregulate monocyte response to CaOx while having no effect upon LPS exposed monocytes. This tolerance mechanism may
Source of Funding: NIH Grant T-32 DK-094789 NIDDK Benign Urology Training Program
MP58-10 CLASSIFICATION OF STONE PATIENTS BY MICRO CT STUDY OF STONES: CORRELATION WITH PAPILLARY PATHOLOGY James C. Williams Jr*, Michael S. Borofsky, Casey Dauw, Andrew P. Evan, Indianapolis, IN; Fredric Coe, Elaine Worcester, Chicago, IL; James E. Lingeman, Indianapolis, IN INTRODUCTION AND OBJECTIVES: Recent endoscopic and biopsy studies with stone formers have suggested that the underlying papillary pathologies differ among stone forming types. Specifically, idiopathic calcium oxalate (CaOx) stone formers who retain stones during their early growth on Randall’s plaque (RP) have no crystal deposition in collecting ducts. In contrast, both apatite and brushite stones are associated with plugging of papillary collecting ducts with apatite mineral. The aim of this study was to extend this work to calcium stone formers in general to see if visual scores of papillary pathology might correlate with morphologies of stones. METHODS: During stone removal procedure, papillary pathology was graded for degree of mineral plugging, pitting of the papilla tip, flattening of papilla shape, and abundance of Randall’s (white) plaque. Papillary scores were 0, 1, or 2 for each measure, and scores were averaged for all papillae in each kidney. All stones removed were photographed, and scanned individually using a Skyscan 1172 micro CT system, with voxel sizes of 2-10 mm, which allowed identification of mineral types and 3D reconstruction of stone structure. Stones were then classified by composition and morphology. CaOx stones that were found to have a concave surface containing apatite (typically showing lumens of calcified tubules and/or vessels, indicating interstitial RP) were classed as stones that had been attached to RP. CaOx stones that
Vol. 195, No. 4S, Supplement, Monday, May 9, 2016
mineral formations appreciated in bone, cementum and dentin. Multiscale analyses of the spatial distribution of organic and inorganic components of biominerals across organ systems may provide insights into the pathogenesis of nephrolithiasis.
e779
partially explain the lack of papillary inflammation in the pathogenesis of Randall’s plaque.
Source of Funding: NIH NIDDK 1P20DK100863 (MLS); NIH NIDCR R01DE022032 (SPH); NIH/NCRR S10RR026645 (SPH)
MP58-09 HYDROXYAPATITE INDUCES CALCIUM OXALATE TOLERANCE IN PRIMARY HUMAN MONOCYTES Paul Dominguez Gutierrez, Sergei Kusmartsev, Benjamin Canales, Vincent Bird, Johannes Vieweg, Saeed Khan*, Gainesville, FL INTRODUCTION AND OBJECTIVES: Although most crystal deposits within tissue produce inflammation, renal interstitial hydroxyapatite deposits most like do not, accumulating as Randall’s plaque. To further explore this lack of response, we investigated the effect of oxalate, hydroxyapatite (HA), and combination of both on time dependent, immunological responses in human monocytes, the precursors to tissue macrophages. METHODS: Primary human monocytes and human monocytic cell line THP-1 cells were exposed to varying concentrations of soluble potassium oxalate (KOx) or insoluble CaOx and HA crystals. Primary human monocytes cells were pre-treated with 100 ug/ml of HA or CaOx followed by secondary treatment with 100 ug/ml HA, 100 ug/ml CaOx, and 1 ug/ml LPS. Cells were collected at various times after various treatments, and RNA was analyzed by quantitative real time PCR. Furthermore, monocytes function to enhance macrophage function and differentiation in the presence of CaOx where observed. RESULTS: Primary monocytes and THP-1 cells responded strongly to CaOx in a dose dependent manner producing TNFa, IL-1b, IL-8, and IL-10 with little to no response to KOx and HA. Pre-exposure to HA had little effect on human monocytes response to CaOx and LPS exposure; however, pre-exposure to CaOx followed by HA negated cytokine production. CONCLUSIONS: CaOx stimulates monocytes to produce cytokines and chemokines which function to stimulate and enhance macrophage uptake of CaOx; furthermore, monocytes exposed to CaOx go on to differentiate into macrophages. Furthermore, CaOx recognition by monocytes appears to be specific. Monocytes exposed to CaOx followed by LPS produce more cytokiens than LPS; however, monocytes exposed to LPS followed CaOx display a greatly downregulate response compared to LPS alone. HA, a component of Randal’s plaques, does not stimulate monocytes; however, it functions to specifically downregulate monocyte response to CaOx while having no effect upon LPS exposed monocytes. This tolerance mechanism may
Source of Funding: NIH Grant T-32 DK-094789 NIDDK Benign Urology Training Program
MP58-10 CLASSIFICATION OF STONE PATIENTS BY MICRO CT STUDY OF STONES: CORRELATION WITH PAPILLARY PATHOLOGY James C. Williams Jr*, Michael S. Borofsky, Casey Dauw, Andrew P. Evan, Indianapolis, IN; Fredric Coe, Elaine Worcester, Chicago, IL; James E. Lingeman, Indianapolis, IN INTRODUCTION AND OBJECTIVES: Recent endoscopic and biopsy studies with stone formers have suggested that the underlying papillary pathologies differ among stone forming types. Specifically, idiopathic calcium oxalate (CaOx) stone formers who retain stones during their early growth on Randall’s plaque (RP) have no crystal deposition in collecting ducts. In contrast, both apatite and brushite stones are associated with plugging of papillary collecting ducts with apatite mineral. The aim of this study was to extend this work to calcium stone formers in general to see if visual scores of papillary pathology might correlate with morphologies of stones. METHODS: During stone removal procedure, papillary pathology was graded for degree of mineral plugging, pitting of the papilla tip, flattening of papilla shape, and abundance of Randall’s (white) plaque. Papillary scores were 0, 1, or 2 for each measure, and scores were averaged for all papillae in each kidney. All stones removed were photographed, and scanned individually using a Skyscan 1172 micro CT system, with voxel sizes of 2-10 mm, which allowed identification of mineral types and 3D reconstruction of stone structure. Stones were then classified by composition and morphology. CaOx stones that were found to have a concave surface containing apatite (typically showing lumens of calcified tubules and/or vessels, indicating interstitial RP) were classed as stones that had been attached to RP. CaOx stones that