D. Bazin , M. Daudon - HÔPITAL NECKER · 2010-05-21 · D. Bazin, Ch. Chappard et al., Osteo. Intern. 2 (2009) 1065-1075. - Revisiting the localisation of Zn2+ cations sorbed on
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Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Nouvelles méthodes d’études des calculs et des plaques de Randall
New techniques of analysis of lithiasis and Randall’s plaque
D. Bazina, M. Daudonb
a Laboratoire de Physique des Solides,Orsay.b AP-HP, Hôpital Necker, Service de Biochimie A, Paris.
Les Actualités NéphrologiquesJean HamburgerHôpital Necker
les lundi 26, mardi 27 et mercredi 28 avril 2010.Maison de la Chimie
28 bis Rue Saint-Dominique - 75007 Paris
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
The ultrastructure of Randall’s Plaque (R’sP )
Epidemiological aspectsR’s P described at the mesoscopic scale through SEMRole of oligoelements in the pathogenesis of R’s P First Ex vivo Synchrotron radiation experiments on R’s P
Application of physical methods to KS charaterisation
Calcium oxalate stonesCalcium phosphate stonesUric acid stones
Conclusion & Perspectives
D. Bazin et al. Ann. Biol. Clin. 64 (2006) 125-139.
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Randall’s plaque (qq 100 µm)
Kidney stone (qq mm)CaC2O4 H2O §§
Crystal 100nmCrystallites qq µm
Randall’s plaque : Definition
§ Epidémiologie, structure et physiopathologieM. Daudon, O. Traxer, D. Bazin, Séminaire d’uronéphrologie 2009.§§ Examination of whewellite kidney stones by SEM and PND techniques.M. Daudon et al., J. of Applied Crystallography 42, 109-115, 2009.§§§ Diffraction techniques and vibrationnal spectroscopy opportunities to characterise bones,D. Bazin et al.,Osteoporosis Int. 20 (2009) 1065-1075.
For a sake of clarity, we used the terms ‘nanocrystals’ and ‘crystallites’ according to Van Meerssche & Feneau-Dupont. In this simplified scheme, a collection of nanocrystals (measuring some hundreds of nanometres) constitutes crystallites (measuring some tens of micrometers).
A. Randall Origin and growth of renal calculi.Ann Surg. (1937) 105 : 1009-1027.
Ca5(PO4)3OH space group is P63/m, a = b= 9.41844 Å, c = 6.88374 ÅNanometer scale crystals§§§
At the surface of attached kidney stones§, a particular deposit termed Randall’s plaque (R’sP) serves as a nucleus.
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Prof. O. Traxer (Tenon Hôpital) : From 2004 to 2007 (436 uretero-renoscopies)289 lithiasic patients et 147 non lithiasic patients (tumeur, sténoses,…)
- Lithiasic: 164 patients with RP (57%)- Non lithiasic : 48 patients with RP (27%)
Fréquence, topographie et classification des PRX. Carpentier, M. Daudon, O. Traxer, D. Bazin, Séminaire d’uronéphrologie 2009.Stone morphology suggestive of Randall's plaque M. Daudon, O. Traxer, P. Jungers, D. Bazin Renal Stone Disease 900, 26-34, 2007.
Epidemiological aspects
Perc
enta
ge o
f KS
with
a R
’sP
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
R’sP described at the mesoscopic scale through SEM
The scanning electron microscope images the sample surface by scanning it with a high energy beam of electrons. Based on the interaction between these incident electrons and atoms at or near the surface sample, this technique is able to give relevant information on local details of the surface at the submicrometer scale.
Experimental aspects
Located at the solid state laboratory, a Zeiss SUPRA55-VP type scanning electron microscope (SEM) is used for microstructure observations. This field effect gun microscope operates at 0.5 to 30 kV. High resolution observations are obtained by two secondary electron detectors : in lens SE detector and Everhart-Thornley SE detector.
To maintain integrity of the kidney stones, measurements have been performed at low voltage (1.4 KeV) and without the usual deposits of carbon at the surface of the sample.
Field Emission Scanning Electron Microscope
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Observation at the mesoscopic scale through SEM
Randall’s plaquesM. Daudon, O. Traxer, J.C. Williams, D. BazinChapitre de livre, Springer-Verlag, in press.
KS
KS
KS
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Interface Papilla/ Randall’s plaque
Interface Randall’s plaque/ Kidney stones
Investigation @ the interfaces
KS
KS
KS
R’s PApatite
R’s PApatite
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Role of oligoelements in the genesis of R’sP
In a previous study, stones were investigated by -Synchrotron Radiation X-ray Fluorescence (-SRXF) at LURE (Laboratoire pour l’Utilisation des Rayonnements Electroniques, Orsay, France) in order to identify and quantify heavy elements
With this experimental set up we can collect several fluorescence spectra for one sample
Zn (ppm)Apatite KS 1059 ± 1056RP 9550 ± 4758Kidney tissue 15-32Urine 0.002-0.1Urine 0.3 ± 0.2
Heavy elements in urinary stones D. Bazin, P. Chevallier, G.; Matzen, M. Daudon, P. Jungers Urological Researc 35, 179-184 , 2007.
Are oligoelements potential risk factors for R’sP and kidney stone formation ? X. Carpentier et al., Submitted to J. of Trace elements in Medicine and Biology
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
In order to obtain a a cartography, we perform PIXE (Protons Induced X-ray Emission) experiments with a 1µm probe
Coll. H. Khodja LPSue (CNRS-CEA)
Role of oligoelements in the genesis of R’s P a PIXE investigation
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Role of oligoelements in the pathogenesis of R’sP -a PIXE investigation
Similar spatial distribution of Ca and Zn
Different spatial distribution of Ca and Zn
At least, two different Zn species on/in R’s P
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
First Ex vivo Synchrotron radiation experiments on R’s P
Dr. X. Carpentier, CHU Nice, Hôpital Pasteur, Nice.PhD thesis end of 2010.
Chemical Composition of R’sPEven if other phosphate phases such as whitlockite or brushite can be found as minor components (less than 5%), calcium phosphate apatite (CA) as well as amorphous carbonated calcium phosphate (ACCP) are the major components of most RPs.
Medical signification : ACCP comes from a calcium phosphate supersaturation with an excess of Calcium and/or phosphate and/or a high pH
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Experiments on Synchrotron Soleil
- Very first tests on SOLEIL regarding the Zn environment in pathological calcifications made of apatite determined by X-ray absorption spectroscopy.D. Bazin, X. Carpentier, O. Traxer, et al., J. of Syn. Rad. 1 (2008) 506-509.http://www.lightsources.org/cms/?pid=1002029- Diffraction techniques and vibrational spectroscopy opportunities to characterise bones D. Bazin, Ch. Chappard et al., Osteo. Intern. 2 (2009) 1065-1075.- Revisiting the localisation of Zn2+ cations sorbed on pathological apatite calcifications made through Xas D. Bazin, X. Carpentier et al., Biochimie 9 (2009) 1294-1300.- The pathogenesis of Randall’s Plaque : A papilla cartography of Ca compounds through an ex vivo investigation based on XANES spectroscopy X. Carpentier, D. Bazin, P. Jungers et al., J. of Syn. Rad. 17 (2010) 374-379.Highlight of Solid State Laboratory in 2010.
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
CAPACC
Characterisation of some points at the tip of the papilla
At either the K- or the L-edge, XANES contains both electronic and structural information, but can be difficult to interpret fully§.
§ Y. Cauchois, N.F. Mott, Philos. Mag. 40 (1949) 126.D.E. Sayers, F.W. Lytle, E.A. Stern, Adv. X-ray Anal. 13(1970) 248.D. Bazin, D. Sayers, J. Rehr J. Phys. Chem. 101 (1997) 11040.D. Bazin, J. Rehr, Catal. Lett. 87 (2003) 85.D. Bazin, J. Rehr, J. Phys. Chem. B 107 (2003) 12398.
4035 4040 4045 4050 4055 4060 4065
B
A
C1C2
Ca K edge
E/eV
Abs
orpt
ion
(a.u
.)
a
b ccd
la
lblcld
Papilla 1
Reference compoundsHAP, CA, PACC
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Peculiar Morphology of stones in primary hyperoxaluriaM. Daudon, P. Jungers, D Bazin, NEJM, 359 (2008)100-101.
Peculiar morphology of stones in primary hyperoxaluria
M. Daudon, C.A.Bader, P. Jungers, Scan. Micro. 7 (1993) 1081.M. Daudon Archives de Pédiatrie 7 (2000) 855.M Daudon, Annales d'Urologie 39 (2005) 209.
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Relationships Between Carbonation Rate of Carbapatite and Morphologic Characteristics of Calcium Phosphate Stones and Etiology
Relationship between the carbonate rate of carbapatite, morphological characteristics of calcium phosphate stones and etiologyX. Carpentier, M. Daudon, O. Traxer, P. Jungers, A. Mazouyes, G. Matzen, E. Véron, D. Bazin, Urology 73 (2009) 968-975.
The presence of bacterial imprints at the surface of KS is well known
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
UA stones, type 2 diabetes & Powder Neutron Diffraction
It is well known that uric acid stones are commonly more prevalent in male stone formers in most western countries.
Recent epidemiologic investigations point out an intimate correlation between obesity, type 2 diabetes and uric acid kidney stone.
Distribution of UA stones with respect to diabetes status in male and female stone former
With diabetes Without diabetesM F M F33.2% 42.5% 11.9% 9.5%
Age 61.4 59.7 59.7 59.6BMI 30.2 33.2 27.1 27.1
M. Daudon,O. Traxer, P. Conort, B. Lacour, P. JungersJASN 17 (2006)2026-2033.
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Coll. G. André, LLB CEA-CNRS
Ø=Kλ/βcos(θ)Ø : Nanocrystal sizeλ : wave lengthβ : Widthθ : position
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Ønanocrystals (F) >> Ønanocrystals (M)
0
50
100
150
200
2 4 6 8 10 12 14 16 18
Non diabetic Male IIIaNon diabetic Male IIIbNon diabetic Female IIIaNon diabetic Female IIIb
Kidney stones
Nan
ocry
stal
s siz
e of
uric
aci
d (n
m)
Non diabetic patients
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Ø (F) >> Ø (M) = Ø (Diab)
0
50
100
150
200
2 4 6 8 10 12 14 16 18
Non diabetic Male IIIaNon diabetic Male IIIbNon diabetic Female IIIaNon diabetic Female IIIb
Kidney stones
Nan
ocry
stal
s siz
e of
uric
aci
d (n
m)
0
50
100
150
200Diabetic Male PatientsDiabetic Female Patients
nano
cris
tal s
ize
of u
ric a
cid
(Å)
Kidney stones of diabetic patients
19 21 23 25 27 2920 22 24 26 28 30
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Conclusion
The ultrastructure of R’sP
- R’P described at the mesoscopic scale through SEM
- oligoelements & R’sP
-First Ex vivo S.R. experiments on R’sP
Application of physical methods to KS charaterization
Calcium oxalate stonesCalcium phosphate stonesUric acid stonesUnderstand the pathogenesis of RP
Use such techniques to perform diagnosticTo explain some epidemiologic data
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
PerspectivesUnderstand the pathogenesis of RP (JSR)Use FESEM to perform diagnostic (NEJM)To explain some epidemiologic data (Submitted)I How drugs dissolve kidney stones: Case of cystine, …
II Using Synchrotron Radiation to perform diagnosticColl. P. Dumas & Ch. Sandt
Tiopronine
http://www.synchrotron-soleil.fr/Soleil/ToutesActualites/2010/Du-synchrotron-a-l-hopitalhttp://www.cnrs.fr/CNRS-Hebdo/paris-michel-ange/actus.php?numero=1882http://www.lewebzine.aphp.fr/spip.php?article408
III Other calcifications (Thyroid§, Cardiac valves§,Cartilage§)
Captopril
§ submitted
Les Actualités Néphrologiques, Maison de la chimie, Avril 2010
Aknowledgments
P.A. Albouy (LPS), D. Thiaudière (Soleil) A. LebailG. André (LLB), S. Reguer (Soleil) P. Dumas (Soleil)Prof. O. Traxer A. Cousson (LLB), Ch. Sandt (Soleil) Prof. I. Tostivint (LPS) Prof. M. Mathonnet (Limoges) Pr. F. Lioté (Lariboisiere) E. Foy (Lab P. Sue), Dr. I. Brocheriou (Tenon) Dr. E. Korn (Lariboisiere) G. Matzen (CEMHTI) Dr. P. Dorfmuller (LPS) Dr. Ch. Nguyen (Lariboisiere)S. Rouzière (LPS), Prof. P. Connort (LPS) Dr. Ch. Chappard (Lariboisiere)E. Véron (CEMHTI) Prof. J.P.Haymann (Tenon) B. Fayard (LPS) R. Weil (LPS) Prof. P. JungersE. Elkaim (Soleil) Ch. Combes (ENSCIAT) Ch. Rey (ENSCIAT) F. Fayon J.M. NedelecH. Khodja LPSue
S. Katelbach, Stage de Pharmacie, Hôpital Necker, Jan-Mai 2005.A. Mazouyes de l'école Centrale Paris & de MII, Paris VI, Mai-Sept 2006.Dr. X. Carpentier, Urologue, Master 2 Sciences et Technologie Mai-Sept 2007.E. Dureault, LIII Licence Physique et application, Un. Paris XI, Mai-Juin 2007.J. Li, LI Parcours PGA de la Licence de Physique, Un. P. et M. Curie, Fév 2008.J. Owen, LI Parcours PGA de la Licence de Physique, Un. P. et M. Curie, Fév 2008.A. Traohé, MI, Ingéniérie et Physique de la Santé, Un. Paris XI, Jan-Mars 2008.Y. Fourdain, Licence LIII Physique et application, Un. Paris XI, Mai-Juin 2008.R. Leguillon, Licence LIII Physique et application, Un. Paris XI, Mai-Juin 2008.A. Perreux, stage BTS, Technicienne de Laboratoire, Juin-Juillet 2008.J. Montanaro, stage BTS, Technicienne de Laboratoire, Juin-Juillet 2008.B. Hannache, stage, Maitre de conférence, Jan 2009Y. Pene, LI PGA de la Licence de Physique, Un. P. et M. Curie, Jan 2009.E. Mukuna, LI PGA de la Licence de Physique, Un. P. et M. Curie, Jan 2009.A.S. Boulon, Licence LIII Physique et application, Un. Paris XI, Mai-Juin 2009. Th. Hamalin, Licence LIII Physique et application, Un. Paris XI, Mai-Juin 2009.S. Passe-Coutrain, Licence LIII Physique et application, Un. Paris XI, Mai-Juin 2009.Dr. Ch. Nguyen, Rhumatologue, Hôpital lariboisiere, Mai-Sept 2009.F. Stephan, stage BTS, Technicienne de Laboratoire, Juin-Juillet 2009.H. Robert, stage BTS, Technicienne de Laboratoire, Juin-Juillet 2009.M. Roméo, stage BTS, Technicienne de Laboratoire, Juin-Juillet 2009.J. Bourgilles, Licence LIII Physique fondamentale, Mai-Juin 2009.C. Moreau, Licence LIII Physique et application, Un. P. et M. Curie, Jan 2010.P. Seznec, Licence LIII Physique et application, Un. P. et M. Curie, Jan 2010.F. Boudaouine, M1 Physique appliquée, Un. Paris XI, Avril-Juin 2010.T. Gou, Licence LIII Physique et application, Un. Paris XI, Mai-Juin 2010.A. Klutsch, Licence LIII Physique et application, Un. Paris XI, Mai-Juin 2010.
3 PhD thesisANR-09-BLAN-0120-02.Ask for a ANR on cartilage (Prof. F. Lioté, Lariboisière) and other programs (Tassili, Un. Paris XI,Toulouse…)
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