AU2022419255B2

AU2022419255B2 - Bone regeneration material

Info

Publication number: AU2022419255B2
Application number: AU2022419255A
Authority: AU
Inventors: Emilie DORY; France LAMBERT; Geoffrey LECLOUX; Eric Rompen
Original assignee: WISHBONE
Current assignee: WISHBONE
Priority date: 2021-12-23
Filing date: 2022-12-22
Publication date: 2025-12-18
Anticipated expiration: 2042-12-22
Also published as: EP4452345B1; ES3054418T3; CN118510554A; KR20240122814A; ZA202404981B; EP4452345A1; BE1030086A1; WO2023118505A1; BE1030086B1; CA3241555A1; JP2024545767A; AU2022419255A1; EP4452345C0

Abstract

The present invention relates to a bone regeneration material consisting essentially of a solid phase of hydroxyapatite of macroporous natural origin, as well as to a method for manufacturing same and to a method for repairing a bone defect.

Description

BONE REGENERATIONMATERIAL BONE REGENERATION MATERIAL

The present The presentinvention inventionrelates relatestotoaabone bone regeneration regeneration material material

in essence in comprising essence comprising a solid a solid phase phase of hydroxyapatite of hydroxyapatite of natural of natural origin origin which is which is macroporous. macroporous.

The present The present invention invention further further relates relatestoto aa method for method for manufacturing bone manufacturing bone regeneration regeneration material material ininessence essencecomprising comprisingaa solid solid

phase phase ofofhydroxyapatite hydroxyapatite of natural of natural origin origin which which is macroporous. is macroporous.

The present The presentinvention inventionfinally finally relates relates to to aa method method forfor repairing repairing

a bone a bonedefect defect in in a patient, a patient, using using bone bone regeneration regeneration material material in essence in essence

comprising comprising aasolid solid phase phaseofofhydroxyapatite hydroxyapatite of of natural natural originwhich origin which is is

macroporous. macroporous.

This type This of bone type of regeneration bone regeneration material material is particularly is particularly used used in in the treatment the treatmentof of bone bone deterioration deterioration in different in different fieldsfields of restorative of restorative or or cosmeticsurgery. cosmetic surgery.

Hydroxyapatite is aa calcium Hydroxyapatite is calciumphosphate phosphate withwith the formula the formula

Ca 5(PO4)3(OH) Cas(PO4)3(OH) with with osteoconductive osteoconductive properties properties and itand it ismain is the the main mineral mineral

component component ofofbone. bone. Indeed, Indeed, hydroxyapatitebelongs hydroxyapatite belongs to to the the crystallographicfamily crystallographic familyof of apatites, apatites, which whichare areisomorphic isomorphic compounds compounds with with the same the hexagonal same hexagonal structure. structure.

This compound This compound hashas been been widely widely usedused as a as a biomaterial biomaterial for for many years many years inindifferent differentmedical medical specialities,since specialities, sincehydroxyapatites hydroxyapatitesareare the the

most common most common crystallinecalcium crystalline calcium phosphates phosphates and and the primary the primary mineral mineral

constituents of constituents of bone, bone,dental dental enamel enamel and and dentin. dentin.

Furthermore, hydroxyapatites and Furthermore, hydroxyapatites andmoremore particularly particularly

hydroxyapatites hydroxyapatites ofof naturalorigin natural originhave have good good biocompatibility biocompatibility and specific and specific

adsorptionproperties adsorption properties forcells for cellsororproteins; proteins;their their use useisis therefore thereforeperfectly perfectly

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suitable in suitable in the field of the field of reconstructive, restorative or reconstructive, restorative or aesthetic aestheticsurgery surgery onon

bone, particularlyoral bone, particularly oral bone. bone.

It isistherefore It therefore recognised that hydroxyapatite recognised that hydroxyapatiteofofnatural natural (animal) origin has (animal) origin hasosteoconductive osteoconductive properties properties as wellasaswell as a crystalline a crystalline

structure and structure anda amorphology morphology identical identical to those to those of a of a natural natural bone bone material material

in humans, in hydroxyapatitebeing humans, hydroxyapatite beingperfectly perfectlysuitable suitable and andcurrently currently being being usedfor used for implants, implants, and andininparticular particular for for intraoral intraoralimplants, implants, to tostimulate stimulatebone bone

reconstruction and reconstruction regeneration atatbone and regeneration bone sites sites with with defects defects or or deterioration. deterioration.

In addition, In it has addition, it has been identifiedthat been identified thata a bone bone regeneration regeneration

material consisting material consisting in in essence ofaasolid essence of solid phase ofhydroxyapatite phase of hydroxyapatiteof of natural natural

origin and origin cleanedofoforganic and cleaned organic substances substances (proteins, (proteins, prions,peptides, prions, peptides, lipids) allowed lipids) allowed for for better better bone regeneration bone regeneration compared compared with awith a solid solid phase phase

of synthetic of synthetic hydroxyapatite. hydroxyapatite.ItItisis ininfact factessential essentialthat that thethe bonebone

regeneration regeneration material, material, when when it isininthe it is theimplanted implanted state state in the in the patient, patient, be be purified of purified of all alltraces tracesofoforganic organic substances soas substances SO asto to promote promoteitsitsintegration integration into the into body,its the body, its osseointegration osseointegrationatatthe the implant implant site,its site, its biocompatibility biocompatibility andSOsothat and thatitit can caninteract interactwith withits its biological biologicalenvironment environment while while avoiding avoiding

unwanted rejectionresponses. unwanted rejection responses.

Bonecolonisation Bone colonisationisisdependent dependent on the on the porous porous characteristics characteristics

of the of the bone regenerationmaterial bone regeneration material and andthe theinterconnection interconnectionbetween betweenitsits

macropores macropores in in number number and size. and size. TheseThese interconnections interconnections form tunnels form tunnels that that allow the allow the passage passageof of cellsand cells and blood blood flowflow between between the pores, the pores, thus thus promoting the formation promoting the formation of of new bone. new bone.

In this In thiscontext, context,document document US5417975 is known US5417975 is fromthe known from theprior prior art, which art, which discloses disclosesthe the product product BioOss® whichis BioOss® which is a boneregeneration a bone regeneration material comprising material comprising aasolid solid phase phaseofofhydroxyapatite hydroxyapatite of of natural natural origin origin

having nanopores, having nanopores,micropores, micropores,and andalso alsopores poresof of diameters diametersgreater greater than than or equal or to 50 equal to 50um. µm.

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Document Document WO2015/049336 WO2015/049336 is also is also known known from from the prior the prior art,art,

whichalso which also discloses discloses aa bone boneregeneration regeneration material material comprising comprising a solid a solid

phaseof phase of hydroxyapatite hydroxyapatiteofofnatural naturalorigin origin having havingpores poreswith withdiameters diameters greater than greater than ororequal equaltoto50 50 um,μm, preferably preferably pores pores withwith diameters diameters of of

between5050and between and 100um. 100 µm.

Unfortunately, although Unfortunately, althoughthese these products products have many have many advantages, their advantages, their mechanical mechanical properties properties could could benefit benefit from from improvement. improvement.

The present The present invention invention aims to address aims to the disadvantages address the disadvantagesofof

the prior the prior art artby by providing providing aa bone regenerationmaterial bone regeneration materialasasmentioned mentioned above,characterised above, characterisedininthat thatsaid saidsolid solidphase phaseof of hydroxyapatite hydroxyapatite is ais a crystalline solid crystalline solidphase of hydroxyapatite phase of hydroxyapatite wherein wherein the the crystals crystals havehave a a size size of between of 20and between 20 and120 120nm, nm,and andhave have a specificsurface a specific surfacearea areaof of between between 8 and 8 20 m²/g. and 20 m2/g.

The bone The boneregeneration regeneration material material according according to the to the present present

inventionis invention is particularly particularly advantageously obtained advantageously obtained after after a sintering a sintering stepstep at at a temperature a of between temperature of between 800°C 800°C and and 1200°C, 1200°C, advantageously advantageously between between

800°C and 800°C and1150°C, 1150°C,preferably preferablybetween between 800°C 800°C andand 1100°C, 1100°C, preferentially preferentially

between 800°C between 800°C and and1050°C, 1050°C, advantageously advantageously between between 800°C 800°C and and

1000°C, particularly advantageously 1000°C, particularly between800°C advantageously between 800°C and and 950°C, 950°C, preferably between preferably between 800°C 800°C and and 900°C, 900°C, advantageously advantageously between 800°C between 800°C and850°C, and 850°C, for for example between example between 810°C 810°C andand 830°C, 830°C, such such as as 820°C. 820°C.

This sintering This sintering step makes step makes it it possible possible in in particular particular to to “weld” "weld"

togetherthe together thecrystals crystalsof of the the solid solid phase ofhydroxyapatite phase of hydroxyapatiteof of natural natural origin, origin,

leading to leading to a a very very large large reduction reduction in innanopores nanopores and micropores, or and micropores, or even even

the complete the disappearance complete disappearance thereof, thereof, andand an an increase increase in the in the sizeofofthe size the crystals as crystals well as as well as the thereduction reductionin in specific specific surface surface area. area. However, However, the the massive reduction massive reduction or or even evenelimination elimination of of nanopores nanoporesand and micropores micropores of of the bone the boneregeneration regenerationmaterial materialis isconsidered considered detrimental detrimental since since

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vascularisationand vascularisation andcolonisation colonisationof of the the material material willnonolonger will longer be be possible, possible,

so that SO that subsequent bone subsequent bone regeneration regeneration risksrisks being being incomplete incomplete or of poorer or of poorer

quality. quality.

Indeed, itit is Indeed, is generally generally recognised recognisedthat thatthethe absence absence of of

microporosity in aa bone microporosity in bone regeneration regeneration material material is detrimental is detrimental to the to the

formation of formation of bone growthand bone growth andosteoconduction. osteoconduction.

Unlike this disadvantage, Unlike this the disadvantage, the sinteringstep sintering step according according to the to the

present invention present invention is is particularly particularly advantageous because advantageous because it makes it makes it it possible to preserve possible to thesurface preserve the surfacetopography topography of the of the regeneration regeneration material, material,

whichpreserves which preservesthethe potential potential for for bone bone regeneration regeneration while reinforcing while reinforcing its its strength. strength.

For For example, the example, the step step of of sintering sintering thethe material material according according to to the present the presentinvention inventionallows allowssubstantially substantiallyspherical sphericalelements elementstoto form form onon thethe

surfaceof surface of the thematerial. material.

Thus, the Thus, material according the material accordingtotothe thepresent presentinvention inventionhas has substantially substantially spherical/ball-like/pseudospherical spherical/ball-like/pseudospherical elements elements of of

hydroxyapatite with hydroxyapatite with a sizeofof(diameter a size (diameteror or equivalent equivalent diameter) diameter) between between

150 and350 150 and 350nm, nm, preferably preferably between between 175 325 175 and andnm, 325 nm, preferentially preferentially

between 200 between 200andand 300 The 300 nm. nm.substantially The substantially spherical/ball- spherical/ball-

like/pseudospherical like/pseudospherical elements elements together together form form a rough a rough surface surface of the of the bone bone regeneration material regeneration material according accordingto to thethe present present invention, invention, unlike unlike thethe

performance performance of of a sintering a sintering step step whose whose temperature temperature is higher is higher than 1200°C than 1200°C

whichwould which would resultininthe result themelting meltingof of the the hydroxyapatite hydroxyapatite onsurface on the the surface of of a material, a material, which whichwould would resultinin ananoverly result overlysmooth smooth surface, surface, which which is is

detrimentaltotothe detrimental thepotential potentialfor forbone bone regeneration. regeneration.

Particularly surprisingly, Particularly surprisingly,the the inventors inventors noticed thatthe noticed that thebone bone regeneration regeneration material material according according to present to the the present invention invention sintered, sintered, that that is is to say to say after after a a sintering sinteringstep, step,and and wherein thecrystals wherein the crystals of of the the hydroxyapatite hydroxyapatite

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phase haveaasize phase have size between 20and between 20 and120 120nm, nm,and and having having a a specificsurface specific surface areaof area ofbetween between 8 and 8 and 20 mis 20 m²/g, 2/g, is equally equally as colonised as colonised bytissue by bone bone as tissue as the methods the methods of of thethe prior prior artart while while having having a stiffer,stronger a stiffer, stronger structure structure andand

having aa rough having rough surface surface topography. topography.

Indeed, the Indeed, the bone bone regeneration regeneration material material according according to to the the present invention present invention is is intended to be intended to beimplanted implanted forfor thethe reconstruction reconstruction

and/or regeneration and/or regeneration of of a bone defect a bone defector or bone bonedeterioration, deterioration, more more particularly in particularly inthe the dental dental field, field,wherein wherein the the forces forces exerted bychewing exerted by chewingare are

significant and significant repeated and repeated over over time. time.

It isistherefore It thereforeparticularly particularlyadvantageous advantageous to to be be ableable to have to have

the bone the boneregeneration regeneration material material according according to thetopresent the present invention invention which which has the has thesame same characteristics characteristics andand advantages advantages asofthose as those of theart the prior prior in art in terms of terms of bone regeneration potential, bone regeneration potential, osseointegration, osseointegration,osteoconduction osteoconduction

while being while stiffer, stronger being stiffer, and stronger andwhose whose surface surface roughness preserves, or roughness preserves, or

evenimproves, even improves, bone boneregeneration regenerationpotential. potential.

The dependent The dependentclaims claims refer refer to to other other advantageous advantageous embodiments. embodiments.

Advantageously, Advantageously, the the crystals crystals of the of the crystalline crystalline solid solid phase phase of of hydroxyapatite ofofthe hydroxyapatite thebone bone regeneration regeneration material material according according to theto the

present invention present invention have havea asize size of of between between30 30 andand 120 120 nm, preferably nm, preferably

between between 4040and and100 100nm, nm,preferentially preferentially between 45and between 45 and8080nm, nm,preferably preferably between between 5050and and8080 nm, nm, more more advantageously advantageously between between 5060 50 and and nm.60 nm.

Advantageously,thethe Advantageously, specific specific surface surface areaarea of solid of the the solid hydroxyapatite hydroxyapatite phase phase of the of the bone bone regeneration regeneration material material according according to the to the

present present invention invention isis between between 10 10 and and 20 20 m2/g, preferably m²/g, preferablybetween 10 and between 10 and

18 18 m 2/g, preferably m²/g, preferably between 12 and between 12 and16 16m2/g. m2/g.

This has This has the the advantage advantage ofof providing providing a bone a bone regeneration regeneration

material according material according to to the the present present invention invention which which offers offers at least at least thethe same same

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characteristics of characteristics of bone regeneration bone regeneration potential potential as the as the methods methods of theofprior the prior art, while art, while having improved having improved strength strength andand stiffness, stiffness, particularlysuited particularly suitedtotothe the dentalfield dental field where mechanical where mechanical constraints constraints are are significant. significant.

Preferably, the bone Preferably, the bone regeneration regeneration material material according according to the to the

present invention present invention has has aaporosity porosity ofofbetween between 70 and 70 and 85%, 85%, preferably preferably

between between 7575and and 85%,preferentially 85%, preferentially between 80and between 80 and85%. 85%.

This has This has the the advantage advantage ofof providing providing a a bone bone regeneration regeneration

material according material accordingtotothethe invention invention having having a significantly a significantly improved improved

potential for bone potential for formation. bone formation.

Preferably, the Preferably, the bone bone regeneration regeneration material material according according to the to the presentinvention present inventionhas has a particle a particle size size distributiondio distribution d10ofofbetween between350 350 and and 500 µm, 500 preferably between um, preferably 370and between 370 and480 480um. µm.

present inventionhas present invention has a particle a particle size size distributiond50 distribution d50ofofbetween between500 500 and and

800 µm, 800 preferably between um, preferably 550and between 550 and780 780um. µm.

Preferably, the Preferably, the bone bone regeneration regeneration material material according according to the to the presentinvention present inventionhas has a particle a particle size size distributiond90 distribution d90ofofbetween between850 850 and and 1250 µm, preferably 1250 um, preferably between between 850 850 and and 1100 1100 um,µm, preferentially preferentially between between

850 and 850 1000 um. and 1000 µm.

invention having invention such aaparticle having such particle size size distribution distribution has the has advantage the of advantage of

having an having an optimal optimal pore pore volume volumeallowing allowingbone bone regeneration. regeneration.

Advantageously, thebone Advantageously, the bone regeneration regeneration material material according according

to the to presentinvention the present inventionisis enriched witha asecond enriched with second synthetic synthetic solid solid phase phase of of

calcium phosphate calcium phosphate having having a Ca/P a Ca/P molarmolar ratioratio of between of between 0.22,and 2, 0.2 and

preferablybetween preferably between0.30.3 andand 1.8,1.8 , preferentially preferentially between between 0.51.65, 0.5 and and 1.65, said said second second synthetic synthetic solidphase solid phase having having a solubility a solubility product product Ks greater Ks greater than than

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thesolubility the solubility product product Ks Ks of of said said firstphase first phase of solid of solid hydroxyapatite hydroxyapatite of natural of natural

origin. origin.

In In a particularly advantageously a particularly manner, advantageously manner, thisensures this ensures an an

adequate adequate release release of of calcium calcium (for(for example example in theinform the form of free of free extracellular extracellular

Ca ions) and Ca2+2+ions) andphosphorus phosphorus (for(for example example in theinform theof form freeofextracellular free extracellular PO 43- ions) PO43- ions) into into the the surroundings of the surroundings of boneregeneration the bone regeneration site site SO so the the latter latter

canact can actasaspromoters promoters of the of the regrowth regrowth of surrounding of surrounding biological biological tissues tissues by by significantly promoting significantly theproliferation promoting the proliferationand and differentiation differentiation of of bone bone cellscells

as well as well as as mineralisation. mineralisation.

Preferentially, Preferentially, the the bone regeneration bone regeneration material material according according to to the present the present invention invention comprises at least comprises at least one one therapeutic agentchosen therapeutic agent chosen from among from amongthe the groupgroup comprising comprising antibiotics, antibiotics, antivirals, antivirals, anti- anti- inflammatories,hormones inflammatories, hormonessuchsuch as steroids, as steroids, growth growth factors factors such such as as BMPs, BMPs, anti-rejection agents, anti-rejection stemcells, agents, stem cells, and andmixtures mixturesthereof. thereof.

Advantageously,the Advantageously, thebone bone regeneration regeneration material material according according

to the to thepresent present invention invention is a is a sterile sterile material. material.

Other embodimentsofofthethe Other embodiments bone bone regeneration regeneration material material according according toto thepresent the present invention invention areare indicated indicated in the in the appended appended claims.claims.

The present The present invention invention also also relates relates to to a amethod method for for

manufacturingthe manufacturing thebone bone regeneration regeneration materialaccording material accordingto to the the present present

invention, involving: invention, involving:

- - bringing bringing a a bone material, containing bone material, hydroxyapatiteand containing hydroxyapatite andorganic organic substances,into substances, intocontact contact with with an an aqueous aqueous extraction extraction solution solution broughtbrought to to a temperature a of between temperature of 150°Cand between 150°C and 300°C 300°C and and to to aa pressureof pressure of between between

1500 kPaand 1500 kPa and 3500 3500 kPakPa SO ,as sotoasobtain to obtain a first a first liquid liquid phase phase containing containing said said

organic substances organic substancesand and possibly possibly impuritiesextracted impurities extractedfrom from said said bone bone

material, anda asecond material, and second phase phase of solid of solid hydroxyapatite, hydroxyapatite,

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- - a separation a separation between between said said liquid liquid phase phase and andsaid said solid solid hydroxyapatite phase, hydroxyapatite phase, - a a(gentle) - (gentle)sintering sinteringofofsaid saidseparated separated solid solid hydroxyapatite hydroxyapatite phase phase at at a temperature a of between temperature of between 800°C 800°C and and 1200°C, 1200°C,

- - said sintered said hydroxyapatite sintered hydroxyapatite phase phase forming forming saidsaid bonebone regeneration regeneration

material. material.

The method The method according according to invention to the the invention makesmakes it possible, it possible, in a in a particularly surprising particularly surprisingand advantageous and manner,totoprovide advantageous manner, providea asintered sintered bone regeneration bone regeneration material material which which has has at at least least the same the same characteristics characteristics in in

terms of terms of bone bone regenerationpotential, regeneration potential, osseointegration osseointegration and and osteoconduction osteoconduction as as thethe prior prior art,while art, whilebybybeing being stiffer, stronger stiffer, stronger and whose and whose

surface roughness surface preserves, or roughness preserves, or even improves, bone even improves, regeneration bone regeneration potential. potential.

In particular, In particular,ititappeared that sintering appeared that sintering conditions below conditions below

800°Cdid 800°C didnot notprovide provide forfor an an increase increase in the in the mechanical mechanical strength strength of the of the material, material, while whiletemperatures temperatures above 1200°C, or above 1200°C, or even above900°C, even above 900°C,had hada a negative impact negative on the impact on the surface surface topography topography and and on onthe thebone bone regeneration potential regeneration potential ofofthe thematerial material while while sintering sintering conditions conditions of of between800°C between 800°C and and 1200°C, 1200C, or even or even lower lower thanthan 900°C, 900°C, make make it possible it possible

to obtain to obtain aasolid, solid, robust, robust, strong strong bone regenerationmaterial bone regeneration materialwith withthe the desired bone desired regenerationpotential bone regeneration potentialto to be beable abletotobebeimplanted implanted in in the the

patient. patient.

Advantageously, the Advantageously, the method methodaccording accordingtotothe theinvention invention further comprises further comprises a a seriesofofsieving series sievingonon a series a series of of sieves, sieves, between between said said

separationstep separation stepand and said said sinteringstep, sintering step,ofofsaid said solid solid hydroxyapatite phase, hydroxyapatite phase,

preferablysaid preferably saidsieving sievingseries series comprises comprisesat at least least a first sieving a first sieving on ona a1 1mmmm sieve and sieve andatatleast leastaasecond second sieving sieving on on a 0.25 a 0.25 mm mm sieve. sieve.

For example, For the example, the solidhydroxyapatite solid hydroxyapatite phase phase afterafter extraction, extraction,

which comes which comesfrom from a bone a bone material, material, is is already already weakened weakened by the by the

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extractionstep extraction stepand andisis deposited deposited onon a set a set ofofsieves sievescomprising comprising from from bottom bottom

to top, to top, a collection basket, a collection basket,aa0.25 0.25mm mm sieve sieve andand a 1a mm1sieve. mm sieve.

Even more Even moreadvantageously, advantageously,thethe sieving sieving series, between series, between the the

separation step separation step and andthe thesintering sintering step, step,of ofthe themethod accordingtotothe method according the

invention, is invention, is aa sieving sieving series series involving involving adding metal adding metal balls balls toto said said seriesofof series

sieves, and sieves, setting said and setting said metal metalballs ballsin in motion motionononsaid saidseries seriesofofsieves. sieves.

Indeed,bybyadding Indeed, adding metal metal balls balls to the to the sieve sieve set,set, which which are are set set in motion in motion using usingequipment, equipment, thethe balls balls willwill furtherweaken further weaken the the hydroxyapatite hydroxyapatite phase phase and and the passage the passage of theof the latter latter through through the sieves the sieves will will

be facilitated be facilitated in in order order to to obtain obtain particles particles of theof the desired desired size. size.

Advantageously,thethe Advantageously, aqueous aqueous extraction extraction solution solution of of the the methodaccording method accordingtotothe theinvention invention is is brought to a brought to a temperature temperature of of between 170°Cand between 170°C and 280°C, 280°C, preferablybetween preferably between 190°C 190°C and and 260°C, 260°C, preferentially preferentially between between 210°C 210°C and and 240°C, 240°C, advantageously between advantageously between 220°C 220°C

and230°C. and 230°C.

Preferably, the Preferably, the aqueous extractionsolution aqueous extraction solution of of the the method method according according to to the the invention invention is is brought brought to to a pressure a pressure of between of between 2000 2000 and and 3500 kPa, 3500 kPa, preferably preferably between 2500and between 2500 and3500 3500kPa, kPa,preferentially preferentially between between

3000 and 3000 3500 kPa, and 3500 kPa, advantageously between advantageously between 3200 3200 andand 3500 3500 kPa, kPa, or or even even

between 3400and between 3400 and 3500 3500 kPa. kPa.

Indeed, the Indeed, thesupercritical supercritical extraction extraction step stepofofthe themethod method according to the according to the invention invention under under temperature conditions of temperature conditions of between 220 between 220

and 230°C and 230°C and andpressure pressure conditions conditions of of between 3200 and between 3200 and3500 3500kPa, kPa, presents thebest presents the bestresults results for for obtaining obtaining aapure pure solidhydroxyapatite solid hydroxyapatite phase, phase,

whichisis cleaned which cleaned of of organic organic substances substances (proteins, (proteins, prions, prions, peptides, peptides, lipids) lipids)

thus reducing thus reducing unwanted unwanted rejection rejection responses responses thatthat could could subsequently subsequently

occur. occur.

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The duration The duration ofofthethe supercriticalextraction supercritical extractionstep stepis is advantageouslyadjusted advantageously adjusted according according to the to the quantity quantity of bone of bone material. material.

Advantageously,the Advantageously, the sinteringstep sintering stepofofthe the method method according according to theto the present invention present invention takes takes place place for foraaperiod periodof ofbetween 40 minutes between 40 and44 minutes and

hours, preferably hours, preferablyfor fora aperiod period of of between between 1 and 1 3 and hours,3 preferentially hours, preferentially between1 1and between and2 2hours, hours, for for example between example between 1 and 1 and 1.51.5 hours. hours.

In addition, In addition, the the sintering sintering step step includes includes a a temperature risesub- temperature rise sub- step combined step combined withwith a heating a heating plate plate step; step; this sintering this sintering step step lastslasts in total, in total,

for example, for for11hour example, for hourand and20 20 minutes. minutes.

Preferably, the Preferably, the sintering sintering step of the step of method the method according according to to the the present invention present invention takes takes place at a place at temperatureofofbetween a temperature between 800°C 800°C andand

1150°C, preferably between 1150°C, preferably 800°C between 800°C andand 1100°C, 1100°C, preferentially preferentially between between

800°C and 800°C and 1050°C, 1050°C, advantageously advantageously between between 800°C 800°C and and1000°C, 1000°C, particularly particularlyadvantageously advantageously between 800°C and between 800°C and950°C, 950°C, preferably preferably

between800°C between 800°C and and900°C, 900°C, advantageously between800°C advantageously between 800°C and and850°C, 850°C, for example for between example between 810°C 810°C and and 830°C. 830°C.

Preferably, the Preferably, the temperature rise step temperature rise step of of the the sintering sintering step step

makes makes itit possible possible to to reach reachthe thetemperature temperature of between of between 800°C800°C and 1150°C, and 1150°C,

advantageouslybetween advantageously between 800°C 800°C and and 850°C, 850°C, for for example example between between 810°C 810°C

and830°C, and 830°C,for for aa period period of of between between2020minutes minutes and and 2 hours, 2 hours, preferably preferably

between2020minutes between minutes and and 1 hour, 1 hour, preferentially between preferentially between 2020 minutes minutes and and

45 minutes, 45 minutes, advantageously between advantageously between 25 25 and and 35 35 minutes. minutes.

For example, For the temperature example, the temperaturerise rise step step makes makesitit possible possible to to reachthe reach thesintering sinteringtemperature temperature of 820°C of 820°C in minutes. in 30 30 minutes.

Preferably, during Preferably, duringthe thetemperature temperature rise rise step, step, the the temperature temperature

incrementbased increment based on time on time is substantially is substantially linear. linear.

This allows This allows a a temperature risestep temperature rise stepwhich which is is not not too too long, long, andand

which can which canbe bereproduced. reproduced.

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Indeed,the Indeed, theinventors inventorsnoted noted that that a temperature a temperature rise step rise step with with

relatively long relatively long durations, durations, especially especially at at temperatures above temperatures above 600°C, 600°C, already already

affectedthe affected thebone; bone; this this is isnot notnecessarily necessarily disadvantageous, disadvantageous, butbemust but must be taken into taken into account when account when determining determining thethe duration duration of of thethe plate plate step step atat

the temperature the of between temperature of 800°Cand between 800°C and1200°C, 1200°C,preferably preferably between between 800°C and 800°C and1150°C, 1150°C,advantageously advantageouslybetween between 800°C 800°C and and 850°C, 850°C, for for example between example between810°C 810°Cand and830°C. 830°C.

In addition, In addition, the theheating heating plate plate stepstep of sintering of the the sintering step step is is carried out carried out at at a temperature of a temperature of between between800°C 800°Cand and 1200°C, 1200°C,

between3030minutes between minutes andand 1 hour 1 hour 30 minutes, 30 minutes, preferentially preferentially between between 45 45 minutes and minutes and 11 hour, hour, advantageously between advantageously between 46 46 minutes minutes and and 58 58 minutes. minutes.

For example, For example, the theheating heating plate plate stepstep at aatsintering a sintering

temperature temperature of of 820°C 820°C takes takes place place for for a period a period of between of between 45 minutes 45 minutes and and 58 minutes. 58 minutes.

In addition, In addition, the the sintering sintering step step of of the the method according method according to the to the

inventionhas invention hasa acertain certainflexibility flexibility with with regard to the regard to thetemperature temperatureand and the the duration of duration of sintering. sintering. However, However, the the temperature setting of temperature setting of the the heating heating

plate step plate stepin in the thesintering sintering step step is is the mostimportant the most important setting setting and and benefits benefits

frombeing from beingmore more strictlycontrolled. strictly controlled.

Advantageously, the Advantageously, the method methodaccording according to to thethe present present inventionfurther invention further comprises, comprises,between between said said sieving sieving step step andsintering and said said sintering step, aa step step, stepofoftreating treatingsaid saidsolid solidhydroxyapatite hydroxyapatite phase phase with peroxides, with peroxides,

preferablyaastep preferably stepofoftreating treatingwith withhydrogen hydrogen peroxide. peroxide.

Preferably, the Preferably, the method according method according to to thethe invention invention further further

comprises one comprises onebetween between said said peroxide peroxide treatment treatment step step andand saidsaid sintering sintering

step, a step, (gentle) drying a (gentle) dryingstep. step.

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Advantageously, the Advantageously, the method methodaccording according to to thethe present present invention further invention further comprises comprises a step of a step of enriching enriching the boneregeneration the bone regeneration material with material withcalcium calciumandand phosphorus, phosphorus, by at by at one least least oneand first firstatand at least least one secondseparate one second separatesoaking soakingsucceeding succeeding oneone another another in any in any order, order, said said

at least at least one first soaking one first soaking taking taking place in a place in a first first solution comprising solution comprising calcium calcium

at a at concentrationof a concentration of 1M 1Mand and saidatatleast said leastone onesecond second soaking soaking taking taking

place in a place in a second solution comprising second solution comprising phosphorus at aa concentration phosphorus at concentrationof of 0.5M. 0.5M.

Advantageously, thefirst Advantageously, the first soaking soaking of of the the enrichment step of enrichment step of

the method the method according according to thetoinvention the invention takesinplace takes place in a a first first solution solution of of Ca(NO3)2.4H2O,of Ca(NO3)2.4H2O, of CaCl2.2H2O, CaCl2.2H2O,of of CaSO4.2H2O CaSO4.2H2O and and of of CaCO3. CaCO3.

Advantageously,the Advantageously, thesecond second soaking soaking ofofthe theenrichment enrichment step step

of the of the method according method according to the to the invention invention takes takes place place in a in a second second solution solution

of Na of 3PO4, of Na3PO4, of Na 2HPO4, of Na2HPO4, of NaH2PO4.H2O, NaH2PO4.H2O,ofofK3PO4, K3PO4,of of K2HPO4, K2HPO4,of of KH2PO4, KH2PO4,

of of K2HPO4,of K2HPO4, of (NH4)3PO4, (NH4)3PO4,ofof(NH4)2HPO4 (NH4)2HPO or4 or of of NH4H2PO4. NH4H2PO4.

Preferably, Preferably, the the method according method according to to thethe invention invention further further

comprises comprises a a step step ofof sterilising the sterilising boneregeneration the bone regeneration material material and/or and/or the the

enrichedbone enriched bone regeneration regeneration material, material, preferably preferably a stepaof step of sterilisation sterilisation by by ionisation. ionisation.

Other Other embodiments embodiments ofofthe themanufacturing manufacturing method method according according

to the to presentinvention the present inventionare areindicated indicatedin in the the appended appended claimsclaims

The present The presentinvention inventionalso alsorelates relatestotoa amethod methodfor for repairing repairing a a bone defect bone defect in in a a patient patient comprising comprising the the steps steps of: of:

- - measuring the measuring the defect defect to to be be filled, filled,

- - positioning positioningaasynthetic syntheticcovering device covering devicearranged arranged to topromote promote bone bone

regeneration of regeneration of a bonedefect, a bone defect,being beingobtained obtained by by additive additive manufacturing, comprisingatatleast manufacturing, comprising least one oneshell shell formed formedofofaaporous porous matrix havingaaseries matrix having series of of pores of size pores of size between between 5050 and and 1000 1000 um,µm and, and

13

at least at least one onesupport support pillarlinked pillar linkedonon thethe oneone handhand to porous to said said porous matrix and matrix resting on and resting the other on the other hand handonona a bone bone surface surface of said of said

bone defect,said bone defect, saidcovering coveringdevice device being being arranged arranged to delimit to delimit a a cavity itself cavity itselfarranged arrangedtotoaccommodate a volume accommodate a volume of bone of bone to beto be

regenerated, regenerated,

- filling of of filling said cavity said formed cavity bybythe formed thecovering coveringdevice device with with a a bone bone

regeneration regeneration material material according according to present to the the present invention. invention.

Advantageously, the Advantageously, the method methodaccording according to to thethe present present inventionis invention is aa non-therapeutic repairand/or non-therapeutic repair and/or is is cosmetic. cosmetic.

Advantageously, the Advantageously, the method methodaccording according to to thethe present present invention is invention is for forthe the repair repairofofaa bone subjectedtotostrong bone subjected strongmechanical mechanical stresses, preferably stresses, preferably aabone bone linked linked to to thethe functioning functioning of masticatory of the the masticatory system. system.

Other embodiments Other embodimentsofofthe themethod method of of repairinga abone repairing bone

defect according defect accordingto to the the present present invention invention are areprovided provided in inthe theappended appended

claims. claims.

Other characteristics, Other characteristics, details detailsand and advantages advantages ofofthe the inventionwill invention will emerge from emerge from thethe description description given given below, below, on a on a non-limiting non-limiting

basis basis and and with with reference reference to tothe thedrawings drawingsand and examples. examples.

Examples: Examples:

Figure 11 depicts Figure an image depicts an imageobtained obtained by by scanning scanning electron electron

microscopy(SEM) microscopy (SEM)of of thethe structure structure of of a bone a bone regeneration regeneration material material

according according toto the the priorart, prior art, sintered sinteredat at aatemperature temperature above above 1200°C. 1200C.

Figures 2A Figures and2B 2A and 2Bdepict depictananimage image obtained obtained by scanning by scanning

electron microscopy electron microscopy(SEM) (SEM)ofofthethe structureofofthe structure thebone bone regeneration regeneration

material according material accordingto to thethe invention, invention, sintered sintered at aattemperature a temperature of 820°C, of 820°C,

respectivelyat respectively at aamagnification magnificationof of 5,000 5,000 times times andand 10,000 10,000 times. times.

14

Example1 1- –Bone Example Bone regeneration regeneration material material according according to to the the presentinvention present invention

A bone A boneregeneration regenerationmaterial material has has been beenproduced produced according according

to the to the present present invention invention which whichconsists consists in in essence of aa solid essence of solid phase phaseofof

hydroxyapatite.InInthis hydroxyapatite. this case, case,the thematerial materialhas hasbeen been sterilised. sterilised.

A series A series of of analyses analyses were werecarried carriedoutout including including thethe determination of the determination of thecomposition compositionof of thethe solidphase, solid phase, thethe size size of of thethe

crystals, the crystals, volumetricporosity, the volumetric porosity,thethe size size of of the the particles, particles, the the specific specific

surfacearea, surface area,for for33 samples samples ofof the the bone bone regeneration regeneration material material according according

to the to presentinvention. the present invention.

Samples1,1,22and Samples and3 3all all have havea asolid solidphase phase composition composition of 100% of 100%

hydroxyapatite.That hydroxyapatite. That isisto tosay, say,the thebone bone regeneration regeneration material material according according

to the to the present present invention invention consists consists ininessence essence of of a solid phase a solid of phase of hydroxyapatite. hydroxyapatite.

Sample1 1has Sample hasa acrystal crystal size size of of 54.6 54.6nm, nm, sample sample 2 2 of of 54.2 54.2 nm nm

andsample and sample3 3ofof54.4 54.4nm. nm.The The average average size size of of the the crystalsof crystals of the the bone bone material according material accordingto to thethe present present invention invention is therefore is therefore 54.4 54.4 nm.nm.

Regarding volumetricporosity, Regarding volumetric porosity, sample sample1 1has has a porosity a porosity of of

82.3%, sample 82.3%, sample 22 of of 82.1% 82.1% and sample33of and sample of 82.5%. 82.5%. The The average volumetric average volumetric

porosity of porosity of the the bone regenerationmaterial bone regeneration materialaccording according to the to the present present

inventionis invention is therefore 82.3%. therefore 82.3%.

Concerningparticle Concerning particlesize, size,sample sample 1 has 1 has a particle a particle size size distribution dd10 distribution 10 of of381 381µm, um, d 50 of d50 of553 553 µm andd90 um and d90of of877 877um. µm.Sample Sample 2 has 2 has a a particle size particle size distribution distributionddio 10 of of452 452µm, um, d 50 of d50 of 782 µmand 782 um andd90d90 of of 1243 1243 um.µm.

Sample Sample 3 3 has has a particle a particle size size distributiondio distribution d10ofof474 474um, µm, d50 d50 ofof 756 756 um µm and and

d90 of d90 of 1115 µm. The 1115 um. Theaverage average particlesize particle sizedistribution distribution d 10 of dio of the the bone bone

regeneration regeneration material material according according to present to the the present invention invention is therefore is therefore 436 436 µm,d50 um, d50 is is 697 697 µm andd90 um and d90isis 1079 1079um. µm.

15

All three All three of of samples 1, 22 and samples 1, and 33 have havea aspecific specificsurface surface area area of of 16 16 m 2/gand m²/g and therefore therefore an an average average specific specific surface surface area area of of 16 m2/g. 16 m²/g.

Furthermore, the bone Furthermore, the boneregeneration regeneration materialaccording material according to to

the present the presentinvention inventionisis obtained obtainedin in a a particularlyadvantageous particularly advantageous mannermanner

after a after sintering step a sintering step at at a temperature a temperature of of 820°C 820°C for for a period a period of between of between

45 minutes 45 minutesand and6060 minutes minutes of of heating heating plate, plate, making making it possible it possible to obtain to obtain a a material having material havinga a stiffer, stronger stiffer, strongerstructure structurewith witha rough a rough surface surface

topography,improving topography, improvingbone bone regeneration regeneration potential. potential.

Indeed, as Indeed, asdepicted depictedin in Figures Figures 2A 2A and and 2B, material 2B, the the material

accordingtotothethe according present present invention invention has substantially has substantially spherical/ball- spherical/ball-

like/pseudospherical elements like/pseudospherical of hydroxyapatite elements of hydroxyapatiteofofaasize size (diameter (diameteroror equivalent diameter) equivalent diameter) between between 150 150 and and 350350 nm,nm, preferably preferably between between 175 175 and325 and 325 nm, nm,preferentially preferentially between between 200 200 and 300 nm, and 300 nm, which whichtogether togetherform form a roughsurface. a rough surface.Unlike Unlikethe thebone bone regeneration regeneration material material depicted depicted in Figure in Figure

11 which which has has been sintered at been sintered ataatemperature temperature above 1200°Cand above 1200°C andwhich whichhas has a smooth a smooth surface, surface, unfavourable unfavourable to the to the potential potential for bone for bone regeneration. regeneration.

Example 22 - - Preparation Example Preparation of of bone boneregeneration regenerationmaterial material according according toto the the present present invention. invention.

A batch A batchofofbone bone regeneration regeneration material material according according to to the the

present invention present invention was was prepared preparedbyby collecting collecting bovine bovine bones bones and and then then cutting the cutting the bovine bovinebones bonesto to form form a bone a bone material. material.

The bone The bonematerial materialcontaining containing hydroxyapatite hydroxyapatite andand organic organic

substanceswas substances was brought brought intointo contact contact with with an aqueous an aqueous extraction extraction solutionsolution

undersupercritical under supercriticaltemperature temperatureand and pressure pressure conditions, conditions, for example for example of of

between 220°C between 220°C and and 230°C 230°C andand at aatpressure a pressure of of between between 32003200 and and 3500 3500

kPa, in kPa, in order order to to obtain obtain a a first first liquid phase liquid phasecontaining containing the the extracted organic extracted organic

substances and substances andimpurities impurities and andaasecond second phase phase of of solidhydroxyapatite, solid hydroxyapatite,

16

these two these two phases phases were werethen thenseparated separatedto to preservethethe preserve solid solid hydroxyapatite phase. hydroxyapatite phase.

The solid The solid hydroxyapatite hydroxyapatite phase, phase, weakened weakened bybythethe supercritical extraction supercritical step, was extraction step, wasplaced placedon on a first1mm a first 1mm sieve sieve to carry to carry out out

a first a first sieving, then sieving, onona asecond then second 0.25mm sievetotocarry 0.25mm sieve carryout outa asecond second sieving. Advantageously, sieving. metalballs Advantageously, metal balls are are placed placedononthe thesieves sieveswith with the the solid hydroxyapatite solid phase hydroxyapatite phase to facilitate to facilitate breaking breaking and and sieving. sieving. The sieving The sieving

steps may steps maytake take place place oneone or more or more times, times, for example for example 2 or 3 2times. or 3 times.

The solid The solid hydroxyapatite phaseafter hydroxyapatite phase after sieving sieving was was collected collected

andsubjected and subjectedtotoaasintering sintering step step at ataatemperature temperature of of between 800and between 800 and 1200°C, ideally at 1200°C, ideally at aa temperature temperatureof of 820°C 820°C during during a heating a heating plate plate step at step at

820°C for 820°C for between 45minutes between 45 minutesand and6060minutes. minutes.

The sintered The sintered solid solidhydroxyapatite hydroxyapatitephase phase thus thus forms forms the thebone bone

regeneration material regeneration material according accordingtotothe thepresent presentinvention, invention,which whichhas hasa a

stiffer structure, stiffer structure,isisstronger stronger and alsohas and also hasa rough a rough surface surface topography, topography,

thereby improving bone thereby improving boneregeneration regenerationpotential. potential.

In addition, In the sintered addition, the sinteredsolid solid hydroxyapatite hydroxyapatite phase phase forming forming

the material the material according accordingto to thethe invention invention can can also also be rinsed be rinsed and/orand/or

enriched with enriched with calcium calcium and andphosphorus phosphorusbybyseparate separate soakings,then soakings, then

sterilised, preferably sterilised, preferably byby ionisation. ionisation.

It isisofofcourse It course understood thatthethe understood that present present invention invention is no is in in no way limited way limited to to the the embodiments embodimentsdescribed describedabove above andand thatthat manymany modifications can modifications can be bemade made without without departing departing from from the scope the scope of theof the appendedclaims. appended claims.

Claims

17 CLAIMS CLAIMS

1. 1. Boneregeneration Bone regenerationmaterial materialconsisting consisting in in essence of a essence of a

solid phase solid of hydroxyapatite phase of hydroxyapatiteofofnatural naturalorigin origin which whichisis macroporous, macroporous, having pores having pores with with diameters diameters greater greater than thanor or equal equalto to 50 50 um, µm,preferably preferably

pores pores with with diameters diameters between between 5050and and 100 100 µm, um, characterised characterised in inthat thatsaid said solid phase solid of hydroxyapatite phase of hydroxyapatite is isa acrystalline crystalline solid solid phase ofhydroxyapatite phase of hydroxyapatite wherein the wherein the crystals crystals have have a a size sizebetween 20 and between 20 and120 120nm, nm,and and said said solid solid

phase phase ofofhydroxyapatite hydroxyapatitehashas a specific a specific surface surface areaarea of between of between 8 and 20 8 and 20

m 2/ g. m²/ g.

2. 2. Bone regeneration Bone regeneration material material according according to to claim claim 1, 1, whereinsaid wherein saidcrystals crystals of of said said crystalline crystalline solid solidphase phase of of hydroxyapatite have hydroxyapatite have

a size a size between 30 and between 30 and 120 120 nm, nm, preferably preferably between 40 and between 40 and100 100nm, nm, preferentially between preferentially between 45 45 and 80 nm, and 80 preferably between nm, preferably 50and between 50 and8080nm, nm, moreadvantageously more advantageously between between 50 and 50 and 60 60 nm. nm.

3. 3. Boneregeneration Bone regenerationmaterial materialaccording accordingtotoclaim claim1 1or or 2, 2, whereinsaid wherein saidspecific specificsurface surface area area of of said said solidphase solid phase of of hydroxyapatite hydroxyapatite is is between between 1010and and2020m²/g, m2/g,preferably preferablybetween between1010 and and 18 18 m2/g, m²/g, preferably preferably

between1212and between and1616 m²/g. m²/g.

4. 4. Boneregeneration Bone regenerationmaterial materialaccording accordingto to any any oneone of of

claims 1 claims 1 to to 3, 3, which which has has a porosity of a porosity of between 70and between 70 and85%, 85%, preferably preferably

between7575and between and 85%,preferentially 85%, preferentially between 80and between 80 and85%. 85%. 5.

5. Bone regenerationmaterial Bone regeneration materialaccording accordingto to any any oneone of of

claims 11 to claims to 4, 4, which hasaaparticle which has particlesize size distribution distribution ddio 10 of ofbetween 350and between 350 and 500 500 µm, preferably between um, preferably 370and between 370 and480 480um. µm.

6. 6. Boneregeneration Bone regenerationmaterial materialaccording accordingto to any any oneone of of claims 11 to claims to 5, 5, which hasaaparticle which has particlesize size distribution distribution dd50 50 of ofbetween 500and between 500 and 800 µm, 800 preferably between um, preferably 550and between 550 and780 780um. µm. 7.

7. Bone regenerationmaterial Bone regeneration materialaccording accordingto to any any oneone of of

claims 11 to claims to 6, 6, which hasaaparticle which has particlesize size distribution distribution dd90 90 of ofbetween 850and between 850 and

18

1250 µm, preferably 1250 um, preferably between between 850 850 and and 1100 1100 um,µm, preferentially preferentially between between

850 and 850 1000 um. and 1000 µm. 8.

8. Boneregeneration Bone regenerationmaterial materialaccording accordingto to any any oneone of of claims 11toto7,7, which claims whichis isenriched enriched with with a second a second synthetic synthetic solid of solid phase phase of

calcium phosphate calcium phosphate having having a Ca/P a Ca/P molarmolar ratioratio of between of between 0.22,and 0.2 and 2, preferably between preferably between 0.3 0.3 andand 1.8,1.8, preferentially preferentially between between 0.51.65, 0.5 and and 1.65, said said

second second synthetic synthetic solidphase solid phase having having a solubility a solubility product product Ks greater Ks greater than than thesolubility the solubility product product Ks Ks of of said said firstphase first phase of solid of solid hydroxyapatite hydroxyapatite of natural of natural

origin. origin.

9. 9. Boneregeneration Bone regenerationmaterial materialaccording accordingto to any any oneone of of claims 11 to claims to 8, 8, comprising at least comprising at least one therapeutic agent one therapeutic agentchosen chosen from from

among among thethe group group comprising comprising antibiotics, antibiotics, antivirals, antivirals, anti-inflammatories, anti-inflammatories,

hormonessuch hormones such asas steroids, growth steroids, growthfactors factorssuch suchasasBMPs, BMPs, anti-rejection anti-rejection

agents,stem agents, stemcells, cells, and andmixtures mixturesthereof. thereof.

10. 10. Boneregeneration Bone regenerationmaterial materialaccording accordingto to any any oneone of of claims1 1toto9,9,which claims whichis ais sterile a sterile material. material.

11. 11. Method for Method for manufacturing manufacturing aa bone boneregeneration regeneration material according material accordingto to anyany oneone of claims of claims 1 to 110, to 10, involving: involving:

- - bringing a bringing a bone material, containing bone material, containing hydroxyapatite hydroxyapatiteand andorganic organic

substances,into substances, intocontact contact with with an an aqueous aqueous extraction extraction solution solution broughtbrought to to a temperature a of between temperature of 150°Cand between 150°C and 300°C 300°C and and to to aa pressureof pressure of between between 1500 kPaand 1500 kPa and 3500 3500 kPakPa , so , SO asas toto obtain obtain a first liquid a first liquid phase containing phase containing said said

organic substances organic substancesand and optionallyimpurities optionally impurities extracted extracted from fromsaid said bone bone material, anda asecond material, and second phase phase of solid of solid hydroxyapatite, hydroxyapatite,

- aa separation - separation between betweensaid saidliquid liquid phase phaseand and said said solid solid hydroxyapatite phase, hydroxyapatite phase,

- - a (gentle) a (gentle) sintering sintering of of said said separated solidhydroxyapatite separated solid hydroxyapatite phase phase at at a temperature a of between temperature of between 800°C 800°C and and 1200°C, 1200°C,

- - said sintered said sintered hydroxyapatite hydroxyapatite phase phase forming forming saidsaid bonebone regeneration regeneration

material. 30 material.

19

12. 12. Methodaccording Method according to to claim claim 11, 11, further further comprising comprising a a series of series of sieving sieving on on aa series series of of sieves, sieves, between said between said separation separation step step and and said sintering said sintering step, step, of of said solid hydroxyapatite said solid phase, hydroxyapatite phase, said said sieving sieving series series

preferablycomprises preferably comprisesat at leasta a least first sieving first sieving on a 11 mm on a sieve mm sieve and and at at least least a a

secondsieving second sieving on on aa 0.25 0.25 mm sieve. mm sieve.

13. 13. Methodaccording Method according to claim to claim 12, 12, wherein wherein saidsaid sieving sieving

series, between series, said between said separation separation stepstep and and said sintering said sintering step,step, is a sieving is a sieving

series comprising series adding comprising adding metal metal balls balls to said to said series series of sieves, of sieves, and and setting setting

said metal said metalballs balls in in motion onsaid motion on saidseries seriesof of sieves. sieves.

14. 14. Methodaccording Method accordingtotoany anyone one of of claims1111 claims oror12, 12, wherein said wherein said aqueous aqueousextraction extractionsolution solution is isbrought brought to toaa temperature of temperature of

between 170°Cand between 170°C and 280°C, 280°C, preferablybetween preferably between 190°C 190°C and and 260°C, 260°C, preferentially between preferentially between 210°C 210°C and and 240°C, 240°C, advantageously between advantageously between 220°C 220°C

and230°C. and 230°C.

15. 15. Methodaccording Method accordingtotoany anyone one of of claims1111toto13, claims 13, wherein said wherein said aqueous aqueous extraction extraction solutionis isbrought solution broughtto to a pressure a pressure of of between2000 between 2000 and and3500 3500kPa, kPa, preferably preferably between 2500 and between 2500 and3500 3500 kPa, kPa, preferentially between preferentially between 3000 3000 and and 3500 3500 kPa, kPa,advantageously between advantageously between 3200 3200

and3500 and 3500kPa. kPa.

16. 16. Methodaccording Method accordingtotoany anyone one of of claims1111toto15, claims 15, whereinsaid wherein saidsintering sintering step steptakes takesplace place foraaperiod for periodofofbetween between 40 minutes 40 minutes

and 44 hours, and hours, preferably preferably for for a period of a period of between between1 1and and 3 hours, 3 hours, preferentially preferentiallybetween between 11 and and2 2hours, hours, for for example examplebetween between 1 and 1 and 1.5 1.5

hours. hours.

17. 17. Methodaccording Method according to to oneone of of claims claims 11 11 to to 16,wherein 16, wherein said sintering said sinteringstep steptakes place takes placeatata atemperature temperature of ofbetween 800°Cand between 800°C and 1150°C, preferably between 1150°C, preferably 800°C between 800°C andand 1100°C, 1100°C, preferentiallybetween preferentially between 800°C and 800°C and 1050° 1050° C, C, advantageously advantageously between 800°C and between 800°C and 1000°C, 1000°C, particularly particularlyadvantageously advantageously between 800°C and between 800°C and950°C, 950°C, preferably preferably

20

between 800°C and between 800°C and900°C, 900°C, advantageously between800°C advantageously between 800°C and and850°C, 850°C, for example for between example between 810°C°Cand 810 and 830°C. 830°C.

18. 18. Method Method according according to any to any one one of of claims claims 12 to12 tofurther 17, 17, further comprisingbetween comprising between said said sieving sieving step step andsintering and said said sintering step, astep, step a of step of

treating said treating said solid solid hydroxyapatite phase hydroxyapatite phase with with peroxides, peroxides, preferably preferably a step a step

of treating of treating with with hydrogen peroxide. hydrogen peroxide.

19. 19. Methodaccording Method accordingtotoclaim claim18, 18,further further comprising comprising betweensaid between saidperoxide peroxidetreatment treatment step step and and said said sinteringstep, sintering step, aadrying drying step. step.

20. 20. Method Method according according to any to any one one of of claims claims 11 to11 tofurther 19, 19, further comprising anadditional comprising an additionalstep stepofofrinsing rinsingsaid saidsintered sintered hydroxyapatite hydroxyapatite phase. phase.

21. 21. 21. Method Method according according to any to any one one of of claims claims 11 to11 tofurther 20, 20, further comprising aastep comprising stepofofenriching enrichingthethe bone bone regeneration regeneration material material with with

calcium and calcium andphosphorus, phosphorus,byby at at leastone least one first and first at least and at least one second one second

separatesoaking separate soaking succeeding succeeding one another one another in any in any order, order, said atsaid atone least least one first soaking first soakingtaking taking place in aa first place in first solution solutioncomprising comprising calcium at aa calcium at

concentration of concentration of 1M 1Mand andsaid saidatatleast least one secondsoaking one second soakingtaking takingplace place in aa second in solutioncomprising second solution comprising phosphorus phosphorus at a at a concentration concentration of of 0.5M. 0.5M.

22. 22. Methodaccording Method accordingtotoclaim claim 21,21, wherein wherein said said first first soaking of soaking of said saidenrichment enrichment stepstep takes takes placeplace in a solution in a first first solution of of Ca(NO3)2.4H2O,of Ca(NO3)2.4H2O, of CaCl2.2H2O, CaCl2.2H2O,of of CaSO4.2H2O CaSO4.2H2O ororof ofCaCO3. CaCO3. 23. 23. 23. Methodaccording Method accordingtotoclaim claim2121oror22, 22, wherein wherein said said second second soaking soaking of of said said enrichment enrichment step step takestakes place place in a second in a second solutionsolution

of Na of 3PO4, of Na3PO4, of Na 2HPO4, of Na2HPO4, of NaH2PO4.H2O, NaH2PO4.H2O,ofofK3PO4, K3PO4,of of K2HPO4, K2HPO4,of of KH2PO4, KH2PO4, of of K2HPO4,of K2HPO4, of (NH4)3PO4, (NH4)3PO4,ofof(NH4)2HPO4 (NH4)2HPO or4 or of of NH4H2PO4. NH4H2PO4.

24. 24. Method Method according according to any to any one one of of claims claims 11 to11 tofurther 23, 23, further comprisinga astep comprising step ofof sterilising the sterilising the bone regeneration bone regeneration material material and/or and/or the the enrichedbone enriched bone regeneration regeneration material, material, preferably preferably a stepaof step of sterilisation sterilisation by by

ionisation. ionisation.

21

25. Method 25. Method 25. of of repairinga a repairing bone bone defect defect in ainpatient a patient comprisingthe comprising thesteps stepsof: of: - - measuring the measuring the defect defect to to be be filled, filled,

regeneration of regeneration of a bonedefect, a bone defect,being beingobtained obtained by by additive additive manufacturing,comprising manufacturing, comprisingatatleast least one oneshell shell formed formedofofa aporous porous matrix having matrix havingaaseries series of of pores of size pores of size between between 5050 and and 1000 1000 um,µm , and , and

bone defect,said bone defect, saidcovering coveringdevice device being being arranged arranged to delimit to delimit a a

cavity cavity itself itselfarranged arrangedtotoaccommodate a volume accommodate a volume of bone of bone to beto be

regenerated, regenerated,

- - filling of of filling said cavity said formed cavity bybythe formed thecovering coveringdevice device with with a a bone bone

regeneration material regeneration material according accordingtotoany anyone one of of claims claims 1 to 1 to 1010 oror

capable capable ofof being being obtained obtained byone by any anyofone of claims claims 11 to 24. 11 to 24.

26. 26. Method according Method according toto claim claim25, 25, being beinga a non- non- therapeutic repair therapeutic repair and/or and/or being being cosmetic. cosmetic.

27. 27. Methodaccording Method according to claim to claim 25 26, 25 or or 26, being being for for the the repair of repair of aa bone subjectedtotostrong bone subjected strongmechanical mechanical stresses, preferably stresses, preferably aa

bone linkedtotothe bone linked thefunctioning functioningof of the the masticatory masticatory system. system.