Nghin cu ch to hydroxyapatit dng gm xp t v trng v vi

Phm Th Sao
Trng i hc Khoa hc T nhin Lun vn ThS Chuyn ngnh: Ha V c; M s: 60 44 25 Ngi hng dn: PGS.TS. o Quc Hng Nm bo v: 2012

Abstract: Nghin cu ch to gm xp HA n pha bng phn ng thu nhit t v trng v vi t nhin. Tm hiu c ch, thng s nhit ng v xc nh cc iu kin (nhit , p sut, thi gian) thch hp ca cc phn ng to HA xp. Kho st, nh gi v so s nh mt s c trng c bn ca sn phm gm xp HA ch to c . Keywords: Ha v c; Hydroxyapatite; V trng; vi Content

T VN 1. Tnh cp thit ca ti Canxi hydroxyapatit (hay cn c gi l hydroxyapatit, vit tt l HA) c cng thc phn t l Ca5(PO4)3(OH) hay Ca10(PO4)6(OH)2. HA hin ang c nghin cu rng ri do c cc c tnh qu gi nh: hot tnh v kh nng tng thch sinh hc cao vi cc t bo v cc m, to lin kt trc tip vi xng non dn n s ti sinh xng nhanh m khng b c th o thi. HA l dng canxi photphat d hp thu nht i vi c th con ngi vi t l Ca/P ng nh trong xng v rng. Vic nghin cu v s dng vt liu sinh hc HA vi mc ch thay th v sa cha nhng khuyt tt ca xng do bnh l v do tai nn ang ngy cng rng ri. Cc ch phm HA nhng kch thc khc nhau c nhiu ng dng. dng bt, HA kch thc nano (20-100 nm) dng lm

thuc v thc phm b sung canxi, tng cng kh nng hp th canxi ca c th, ngn nga v iu tr bnh long xng. dng mng, mt lp HA mng, siu mn c th to nn lp men rng, cc chi tit ni xng v lp ph b mt cho xng nhn to. HA dng khi xp c th dng in y cc hc rng su v cc vt rn nt xng t nhin, lm xng nhn to m khng b c th o thi. Gn y, ngi ta cn pht hin dng HA xp t nhin da trn khung xp sn c ca mai mc, san h, v trng, vi c kh nng tng thch tt vi c th c cy ghp. Chng cn c kh nng vn chuyn v phn tn mt s loi thuc nh insulin, vitamin, bo v c dc phm trnh khi tng tc vi tc nhn khc trn ng vn chuyn trong c th, gp phn nng cao hiu qu s dng thuc Vit Nam, vic nghin cu, s dng vt liu HA cho mc ch y sinh hc cha c quan tm nhiu. T nm 2005, Phng Ho V c (Vin Ho hc, Vin Khoa hc v Cng ngh Vit Nam) thc hin mt s ti nghin cu tng hp HA bt v HA xp. Trn c s cc kt qu t c, gp phn to ra mt loi vt liu, nh hng ng dng trong dc hc v y sinh hc, c bit l trong phu thut chnh hnh nc ta, chng ti la chn ti: Nghin cu ch to hydroxyapatit dng gm xp t v trng v vi . 2. Tnh hnh nghin cu y khng phi l ti mi. Tc gi thc hin ti trn c s cc cng trnh c cng b trc y ti phng Ha V c, Vin Ha hc, Vin KH&CN Vit Nam nh: - V Duy Hin, o Quc Hng, Phan Th Ngc Bch (2007), Tng hp v kho st nh hng ca nhit n kch thc ht hydroxyapatit bng phng php kt ta ho hc, Tp ch Ho hc, 45(6A), tr.21-25. - V Duy Hin, o Quc Hng, Phan Th Ngc Bch (2008), Nghin cu ch to HA t khung xp t nhin ca mai mc bng phn ng thy nhit , Tp ch Ha hc, 46(2A), tr.118-123.

- V Duy Hin (2010), Nghin cu ch to v c trng ha l ca hydroxyapatit dng gm xp c kh nng ng dng trong y sinh hc, Lun n Tin s Ha hc, Vin Ho hc - Vin Khoa hc v Cng ngh Vit Nam. - o Quc Hng, Phan Th Ngc Bch (2007), Tng hp bt hydroxyapatit kch thc nano bng phng php kt ta ho hc, Tp ch Ho hc, 45(2), tr.147-151. ti ca tc gi cung cp mt hng mi cho vic nghin cu ch to vt liu HA bng phng php thy nhit t hai nguyn liu l v trng v vi. 3. Mc ch v nhim v nghin cu Trn c s mc tiu nghin cu ch to HA dng gm xp, ti tp trung vo nhng vn sau: - Nghin cu ch to gm xp HA n pha bng phn ng thu nhit t v trng v vi t nhin; - Tm hiu c ch, thng s nhit ng v xc nh cc iu kin (nhit , p sut, thi gian) thch hp ca cc phn ng to HA xp; - Kho st, nh gi v so snh mt s c trng c bn ca sn phm gm xp HA ch to c. 4. Phng php nghinn cu Lun vn s dng cc phng php vt l hin i kho st, nh gi cht lng sn phm thu c - Phng php nhiu x tia X - Phng php ph hng ngoi FTIR - Phng php hin vi in t qut SEM - Phng php phn tch nhit vi sai DTA - Phng php phn tch nhit trng-lng TGA 5. Nhng ng gp mi ca lun vn

Lun vn a ra mt hng mi trong nghin cu ch to vt liu HA nc ta l s dng hai nguyn liu v trng v vi. y l nhng vt liu sn c v r tin. 6. B cc ca lun vn Chng 1: Tng quan Chng 2: Thc nghi m Chng 3: Kt qu v tho lun Kt lun chung.

CHNG 1: TNG QUAN

1.1. CC TNH CHT QUAN TRNG CA HYDROXYAPATIT 1.1.1 Tnh cht vt l Hydroxyapatit (HA): Ca10(PO4)6(OH)2; Khi lng phn t: 1004,60 g; T trng ring: 3,156 g/cm3; Nhit nng chy: 1760oC; Nhit si: 2850oC; Tch s tan: 2,12.10-118. Tinh th HA c th tn ti dng hnh que, hnh kim, hnh vy, hnh si hoc hnh cu, vi mu trng, trng ng, vng, nu hoc xanh l... ph thuc vo cc iu kin hnh thnh trong t nhin hoc tng hp khc nhau [51]. C th nhn bit c cc dng tn ti ca tinh th HA nh s dng phng php hin vi in t qut (SEM) hoc hin vi in t truyn qua (TEM) (hnh 1.1).

Hnh 1.1: Cc dng tinh th HA [21] a - hnh kim; b- hnh cu; c - hnh vy; d - hnh que 1.1.2. Tnh cht ho hc Cng thc cu to ca phn t HA c th hin trn hnh 1.3. Phn t HA c cu trc mch thng, cc lin kt Ca-O l lin kt cng ho tr. Hai nhm OH c gn vi nguyn t P hai u mch [55].
-

Hnh 1.3: Cng thc cu to ca phn t HA 1.1.3. Tnh cht sinh hc Vt liu HA c tnh tng thch sinh hc cao vi cc m v cc t bo, d lin kt trc tip vi xng non v khng b c th o thi. 1.2. CC NG DNG CA HYDROXYAPATIT Cc dng HA cc kch thc khc nhau c nhiu ng dng trong y sinh hc v phu thut chnh hnh nh ch to rng gi, sa cha cc khuyt tt ca xng, lm mt gi, lm cht truyn dn thuc... 1.3. CC PHNG PHP CH TO HYDROXYAPATIT Vic nghin cu ch to cc ch phm t vt liu HA cc dng khc nhau c tin hnh t lu trn th gii v t c nhng thnh tu 5

ng k. Tu thuc vo mc ch ng dng, HA cc dng khc nhau c th c ch to bng nhiu phng php t cc nguyn liu khc nhau. Da vo iu kin tin hnh phn ng, c th phn chia thnh: phng php ha hc v phng php vt l. 1.3.1. Phng php ha hc 1.3.1.1. Phng php t y l phng php to pha rn HA t dung dch cha cc nguyn liu ban u khc nhau, bao gm: phng php kt ta, phng php sol -gel hay phng php kt tinh t dung dch bo ho... Cc phng php ny thng c dng tng hp HA dng bt vi kch thc khc nhau. 1.3.1.1.1. Phng php kt ta 1.3.1.1.2. Phng php sol-gel 1.3.1.1.3. Phng php siu m ho hc 1.3.1.1.4. Phng php phun sy 1.3.1.1.5. Phng php in ho 1.3.1.1.6. Phng php phn ng thu nhit 1.3.1.1.7. Phng php ch to vt liu compozit 1.3.1.2. Phng php kh 1.3.1.2.1. Phng php ho-c 1.3.1.2.2. Phng php phn ng pha rn 1.3.2. Cc phng php vt l 1.4. TNH HNH NGHIN CU CH TO V NG DNG CA GM XP HYDROXYAPATIT TRN TH GII V TRONG NC Vic nghin cu ch to vt liu gm HA trn th gii c tin hnh t lu v t c nhng thnh tu to ln, to ra nhng sn phm a dng cho cc ng dng trong y sinh hc. n nay, cng vi cng ngh v k thut tin tin, nhn chung tnh tng thch sinh hc, xp v phn b cu

trc xp ca gm HA c cc nh khoa hc nghin cu chi tit v c nhiu phng php iu chnh theo mong mun. nc ta, vic nghin cu ch to cc hp cht v c c kh nng ng dng lm vt liu sinh hc ni chung v HA ni ring cn cha c ch nhiu. Nm 2003, Vin Cng ngh X him s dng cng ngh ca Italia, bng cch nhng tm khung xp xenlulo vo dung dch huyn ph HA, sau sy v thiu kt to gm. Sn phm ny c th nghim thnh cng bc u trn ng vt. Khoa Ho hc (i hc Bch khoa H Ni) tin hnh nghin cu tng hp HA dng bt v dng mng. T nm 2005 n nay, Phng Ho V c (Vin Ho hc, Vin Khoa hc v Cng ngh Vit Nam) cng b mt s kt qu tng hp HA bt kch thc nano v HA xp bng cc phng php khc nhau. Tuy vy, vic nghin cu ch to HA ni chung v HA c cu trc xp t nhin v cu trc xp nhn to cha c cp y v c h thng.

CHNG 2: THC NGHIM

2.1. DNG C, THIT B V HO CHT 2.1.1. Dng c v thit b H thit b phn ng gm: cc teflon, l nung, thit b thy nhit, bnh n p v cc thit b ph tr (hnh 2.1).

Hnh 2.1: S nguyn l ca h thit b phn ng thu nhit 2.1.2. Ho cht - Mui (NH4)2HPO4 c tinh khit PA (Trung Quc); - Dung dch 25% NH3 trong nc (PA); - V trng (c hm lng CaCO3 dng canxit khong 97-99%) c bc sch lp mng, ct thnh nhng ming nh kch thc 1x1 mm. Sau , c un si nhiu ln bng nc ct v etanol loi b cc tp cht c hc v hu c, ri sy kh nhit 100oC trong 24 gi. - vi t nhin (c hm lng CaCO3 dng canxit khong 97-99%) c nghin thnh cc vin nh, kch thc khong 1x1x1 mm. Sau , un si nhiu ln bng nc ct v v etanol loi b cc tp cht c hc v hu c, ri sy kh nhit 100oC trong 24 gi.

2.2. QUY TRNH TIN HNH PHN NG Vic ch to HA t v trng v vi c tin hnh theo 2 hng: - Hng phn ng trc tip dng CaCO3; - Hng phn ng gin tip qua giai on trung gian phn hu CaC O3 ca v trng ( vi) thnh CaO. S nguyn l ca h thit b phn ng thu nhit nh hnh 2.1. Cc cht phn ng c a vo cc teflon. p sut ca h c iu chnh bng kh nit thng qua bnh n p. nhit cao, phn ng thu nhit din ra theo phng trnh: - Trong phn ng trc tip:
10CaCO3 + 6(NH4)2HPO4 + 2H2O Ca10(PO4)6(OH)2 + 6(NH4)2CO3 + 4H2CO3

(2.1)

- Trong phn ng gin tip:

10CaO + 6(NH4)2HPO4 + 4H2O Ca10(PO4)6(OH)2 + 12NH4OH (2.2)

y l phn ng d th gia hai pha lng-rn, sn phm ph ca phn ng l kh NH3, H2O v CO2. phn ng xy ra theo chiu thun (hng to ra HA), kh NH3 v H2O to thnh sau phn ng c hp th bng axit H2SO4 m c t trong bnh n p. Ngoi ra, vic tin hnh phn ng trong bnh kn v p sut cao cng c tc dng ngn cn s phn hu ca mui (NH4)2HPO4 trc khi tham gia phn ng vi CaCO3 (CaO). Trong phng php thu nhit, phn ng c thc hin trong mi trng nc nhit v p sut cao nhm thc y nhanh phn ng d th. Nc trong phn ng d th c cc chc nng lm mi trng truyn p sut nhm to nng lng cho phn ng v lm dung mi ho tan mt phn cht phn ng. Do , phn ng d th s xy ra b mt phn cch gia pha lng v rn. Cc th nghim c tin hnh trong thi gian, nhit v p sut khc nhau xc nh iu kin thch hp ca phn ng. Cc sn phm sau phn ng c xc nh mt s c trng c bn bng cc phng php: nhiu x tia X (XRD), hp th hng ngoi (FTIR), phn tch nhit (DTA, TGA) v hin vi in t qut (SEM). 9

- Phng php phn ng trc tip Cn 27,78 g CaCO3 (t v trng, vi) v 22,67 g tinh th (NH4)2HPO4 nh tnh ton theo phng trnh 2.1. Hn hp rn ny c a vo cc teflon, cho thm khong 1 ml nc ct. B sung mt lng nh dung dch NH3 trong nc m bo pH ca dung dch l 9-10. Phn ng thy nhit c tin hnh nhit 200 oC, p sut 14 atm, thi gian phn ng 96 v 135 gi. Cc sn phm sau phn ng c kho st bng cc phng php nu. - Phng php phn ng gin tip V trng ( vi) sau khi lm sch c nung nhit 900 oC trong 1 gi m bo CaCO3 phn hu hon ton thnh CaO theo phng trnh: CaCO3 CaO + CO2 (2.3)

Tin hnh phn ng thu nhit ngay sau khi nung gi nguyn cu trc xp ca CaO. Cn 15,55 g CaO v 22,67 g tinh th (NH4)2PO4 c tnh ton theo phng trnh 2.2. Hn hp rn ny c a vo cc teflon, thm khong 1ml nc. Kho st cc yu t thi gian, nhit , p sut v lng nc xc nh iu kin phn ng thch hp. Cc sn phm sau phn ng cng c xc nh bng cc phng php nu. Mt s c trng c bn (XRD, FTIR, TGA, SEM) ca vt liu sau phn ng c so snh vi mu HA chun ca Vin Tiu chun v Cng ngh Quc gia M (NIST) [20]. 2.3. CC PHNG PHP NGHIN CU C TRNG VT LIU HYDROXYAPATIT 2.3.1. Phng php nhiu x tia X (XRD) 2.3.2. Phng php hng ngoi (FTIR) 2.3.3. Phng php hin vi in t qut (SEM) 2.3.4. Phng php phn tch nhit

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2.3.4.1. Phng php phn tch nhit vi sai (DTA) 2.3.4.2. Phng php phn tch nhit-trng lng (TGA) CHNG 3: KT QU V THO LUN 3.1. C TRNG CA V TRNG V VI T NHIN 3.1.1. Mt s c trng ca v trng Khung xp ca v trng cha n 97-99% khi lng l CaCO3.

a Hnh 3.1: c trng XRD (a) v

c trng nhit-trng lng TGA (b) ca v trng

Hnh 3.2: nh SEM ca v trng vi phng i khc nhau nh SEM (hnh 3.2) ca v trng cho thy chng c cc lp mng xp chng kht ln nhau, gia cc lp ny khng c bin r rt v khng sp xp

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theo mt trt t nht nh. B mt v trng li lm v c mt s l xp khng c hnh dng c trng, phn b khng u, kch thc thay i t 50 n 100 nm. 3.1.2. Mt s c trng ca vi t nhin

a Hnh 3.3: Gin XRD (a)

v c trng nhit-trng lng TGA (b) ca vi Gin XRD (hnh 3.3a) cho thy vi cng c thnh phn l CaCO3 dng canxit bn nhit ging nh v trng.

Hnh 3.4: nh SEM ca vi ti cc v tr khc nhau nh SEM (hnh 3.4) cc v tr khc nhau th hin vi c cu trc gm cc lp xp chng kht ln nhau, gia cc lp ny khng c bin v sp

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xp khng theo mt trt t nht nh. Khc vi v trng, b mt cc lp ca vi c kht, kh bng phng, khng c khe h hoc l xp. iu ny c th gy kh khn cho phn ng thy nhit trc tip. 3.2. PHN NG THY NHIT CH TO HA TRC TIP T CaCO3 Phn ng thu nhit trc tip t CaCO3 din ra theo phng trnh 2.1:
10CaCO3 + 6 (NH4)2HPO4 + 2H2O Ca10(PO4)6(OH)2 + 6(NH4)2CO3 + 4H2CO3

Phn ng c tin hnh nhit 200 oC, p sut 14 atm trong thi gian 96 gi

Hnh 3.5: Gin XRD ca mu nhn c sau phn ng trc tip t v trng (a) v vi (b) trong 96 gi Kt qu XRD ca hai mu (hnh 3.5) cho thy, c hai trng hp, sn phm thu c ch yu vn l CaCO3 dng canxit, mc d thi gian phn ng ko di n 96 gi. Song song vi th nghim trn, phn ng trc tip gia v trng ( vi) nghin thnh bt mn vi tinh th (NH4)2HPO4 c tin hnh vi thi gian n 135 gi.

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Hnh 3.6: Gin XRD ca mu thu c sau phn ng trc tip ca v trng (a) v vi (b), thi gian 135 gi

Kt qu XRD ca hai mu (hnh 3.6) cho thy vn ch hnh thnh mt lng HA rt nh sau phn ng vi v trng. Cn vi vi th hon ton khng xut hin HA. Nh vy, c th kt lun rng do CaCO3 dng canxit vi cu trc c kht ca v trng v vi t nhin nn khng ch to c HA bng phn ng thu nhit trc tip trong iu kin p sut 14 atm v 200 oC vi thi gian n 135 gi. Do , chng ti la chn phng php tin hnh phn ng gin tip qua giai on trung gian phn hu CaCO3 thnh CaO. 3.3. PHN NG THY NHIT CH TO HA THNG QUA HP CHT TRUNG GIAN CaO

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3.3.1. Mt s c trng ca v trng v vi sau khi phn hu

Hnh 3.7: nh SEM ca CaO t v trng (a) v vi (b) Hnh 3.7 l nh SEM ca CaO thu c t v trng (a) v vi (b) sau khi nung phn hu 900oC trong 1 gi. Cu trc ca v trng v vi thay i r rt, tr nn xp hn rt nhiu. iu ny gip phn ng thu nhit to HA thng qua CaO c th din ra d dng hn phn ng trc tip. Phn ng thy nhit xy ra gia CaO vi tinh th (NH4)2HPO4 din ra theo phng trnh tng qut 2.2 sau: 10CaO + 6(NH4)2HPO4 + 4H2O Ca10(PO4)6(OH)2 + 12NH4OH 3.3.2. Xc nh iu kin phn ng thch hp 3.3.2.1. Ch to HA xp t v trng bng phn ng gin tip - Xc nh thi gian phn ng Hnh 3.9 l gin XRD ca cc mu nhn c khi thc hin phn ng theo phng trnh 3.1, iu kin: nhit 200 oC, p sut 14 atm trong thi gian 8, 15, 24 v 48 gi.

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b a b

V-8 h

V-15 h

V-24 h

V-48 h

Hnh 3.9: Gin XRD ca cc mu nhn c t v trmg sau phn ng gin tip vi thi gian tng hp khc nhau Tinh th ca HA kt tinh tt hn theo thi gian v sau 24 gi cc vch c trng gn nh tch bit hon ton. Gin XRD ca HA nhn c vi thi gian phn ng 24 gi v 48 gi khng c s khc bit nhau nhiu. T gin XRD c th tnh ton c kch thc ht trung bnh v tinh th ca cc ht HA t v trng theo cc cng thc 2.3 v 2.4. Cc kt qu c trnh by trong bng 3.1. Bng 3.1 : nh hng ca thi gian phn ng n kch thc ht trung bnh v tinh th ca cc mu t v trng Thi gian 15 gi 24 gi d (nm) theo Scherrer 36,7 63,7 tinh th (%) 47 71

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48 gi

70,82

75

Nh vy, kch thc ht trung bnh v tinh th ca cc mu HA t v trng tng mnh khi thay i thi gian phn ng t 15 n 24 gi. Ko di thm thi gian phn ng t 24 ln 48 gi khng lm kch thc ht trung bnh v tinh th tng nhiu. - Xc nh p sut phn ng Th nghim c thc hin trong trong khong p sut t 6 -14 atm.

V-6 atm

V-10 atm

V-12 atm

V-14 atm

Hnh 3.10: Gin XRD ca cc mu tng hp t v trmg vi thi gian 24 gi p sut khc nhau Hnh 3.10 l gin XRD ca mu HA xp tng hp bng phn ng thu nhit gin tip t v trng nhit 200oC, thi gian phn ng l 24 gi vi p sut thay i 6, 10, 12 v 14 atm. Khi p sut tng n 12-14 atm, cc ion PO43- v OH- d dng khuch tn qua lp bin lng-rn phn ng vi CaO, to thnh HA. ng thi, nhit 200oC v p sut cao (12 -14 atm) ngn cn qu trnh bay hi ca nc, dn n phn ng thu nhit xy ra hon ton, tinh th ca HA kt tinh tt hn so vi p sut thp. 17

Gin XRD ca HA nhn c khi phn ng p sut 12 v 14 atm khng c s khc bit nhiu.

Bng 3.2 : nh hng ca p sut phn ng n kch thc ht trung bnh v tinh th ca cc mu t v trng p sut 12 atm 14 atm d (nm) theo Scherrer 57,9 72,6 tinh th (%) 72 74

Kt qu tnh ton trn bng 3.2 cho thy kch thc ht trung bnh tng mnh (t 57,9 ln 72,6 nm), cn tinh th tng nh (t 72 n 74%) khi tng p sut phn ng t 12 ln 14 atm. Nh vy, iu kin nhit 200oC, thi gian 24 gi v p sut 12-14 atm l thch hp phn ng thu nhit xy ra hon ton. - Xc nh nhit phn ng Thc hin cc phn ng p sut 14 atm, thi gian 24 gi vi cc iu kin nhit 150, 200 v 250oC. Hnh 3.11 l gin XRD ca cc mu t v trng tng hp cc nhit khc nhau.

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V-150 C

V-200oC

V-250oC

Hnh 3.11: Gin XRD ca cc mu t v trng tng hp cc nhit khc nhau Gin ca mu tng hp 200 v 250 oC cho thy mu kt tinh tt. Kt qu tnh ton kch thc ht trung bnh v tinh th ca cc mu di nh hng ca nhit c trnh by trong bng 3.3. Bng 3.3: nh hng ca nhit phn ng n kch thc ht trung bnh v tinh th ca cc mu t v trng Nhit 150 oC 200 oC 250 oC d (nm) theo Scherrer 32,1 65,7 73,4 tinh th (%) 52 72 76

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Kch thc ht trung bnh v tinh th tng mnh khi tng nhit t 150 n 200oC, cn trong khong 200 -250oC, cc i lng trn khng tng nhiu. Nh vy, c th kt lun phn ng ch to HA xp t v trng xy ra thun li 200oC, thi gian 24 gi v p sut 12-14 atm. 3.3.2.2. Ch to HA xp t vi bng phn ng gin tip - Xc nh thi gian phn ng Hnh 3.12 l gin XRD ca cc mu nhn c khi thc hin phn ng theo phng trnh 3.1, iu kin: nhit 200 oC, p sut 14 atm trong thi gian 8, 15, 24 v 48 gi.

D-8 h

D-15 h

D-24 h

D-48 h

Hnh 3.12: Gin XRD ca cc mu nhn c t vi sau phn ng gin tip vi thi gian tng hp khc nhau Thi gian 24 gi v 48 gi phn ng xy ra hon ton. Ging nh phn ng ca v trng, trn gin XRD ch tn ti nhng vch c trng ca HA, vi cc vch c cng ln ti 2 l 31,77 v 32,20 o. 20

Kt qu tnh ton kch thc ht trung bnh v tinh th ca sn phm c trnh by trong bng 3.4. Bng 3.4: nh hng ca thi gian phn ng n kch thc ht trung bnh v tinh th ca cc mu t vi Thi gian 24 gi 48 gi d (nm) theo Scherrer 150,7 211,4 tinh th (%) 72 74

Theo tnh ton trn bng 3.4, kch thc ht trung bnh tng mnh, cn tinh th khng c thay i nhiu khi tng thi gian phn ng t 24 n 48 gi. Nh vy, c th kt lun vi thi gian phn ng 24 gi nhn c sn phm HA n pha. - Xc nh p sut phn ng Th nghim c thc hin trong trong khong p sut t 6 -14 atm. D-6 atm D-10 atm

D-12 atm

D-14 atm

Hnh 3.13: Gin XRD ca cc mu tng hp t vi vi thi gian 24 gi p sut khc nhau 21

Hnh 3.13 l gin XRD ca mu HA xp tng hp bng phn ng thu nhit t vi nhit 200oC, thi gian phn ng l 24 gi vi p sut thay i l 6, 10, 12 v 14 atm. p sut 12 v 14 atm, phn ng xy ra hon ton, trn gin ch c cc vch c trng ca HA. p sut 14 atm, cc vch c trng ti 2 bng 31,77 v 32,20o c cng cao hn so vi 12 atm. Kt qu tnh ton kch thc ht trung bnh v tinh th ca sn phm di nh hng ca p sut c trnh by trong bng 3.5. Bng 3.5: nh hng ca p sut phn ng n kch thc ht trung bnh v tinh th ca cc mu t vi p sut 12 atm 14 atm d (nm) theo Scherrer 156,9 187,4 tinh th (%) 68 75

Theo bng 3.5, khi tng p sut phn ng t 12 ln 14 atm, kch thc ht trung bnh v tinh th ca sn phm HA xp khng thay i nhiu. Nh vy, iu kin nhit 200oC, thi gian 24 gi v p sut 14 atm l thch hp phn ng thu nhit ca vi xy ra hon ton. - Xc nh nhit phn ng Thc hin cc phn ng p sut 14 atm, thi gian 24 gi vi cc nhit l 150, 200 v 250oC.

22

D-150oC

D-200oC

D-250oC

Hnh 3.14: Gin XRD ca cc mu t vi tng hp cc nhit khc nhau Hnh 3.14 l gin XRD ca cc mu t vi tng hp cc nhit khc nhau. Kt qu tnh ton kch thc ht trung bnh v tinh th ca sn phm di nh hng ca nhit c trnh by trong bng 3.6. Bng 3.6: nh hng ca nhit phn ng n kch thc ht trung bnh v tinh th ca cc mu t vi Nhit 150 oC 200 oC 250 oC d (nm) theo Scherrer 56,7 170,8 178,4 tinh th (%) 51 73 76

23

Khi tng nhit t 150 ln 200oC, kch thc d v tinh th tng mnh (56,7 ln 170,8 nm; 51 ln 73%). Trong khong 200 -250oC, kch thc d v tinh th khng thay i nhiu (170,8 ln 178,4 nm; 73 ln 76%). 3.3.3. Mt s c trng ca HA xp t phng php gin tip 3.3.3.1. Ph FTIR

b Hnh 3.15: Ph FTIR ca mu HA thu c t v trng (a) v vi (b) sau phn ng gin tip 200oC, 14 atm v 24 gi Hnh 3.15a l ph FTIR ca mu HA t v trng. Cc vch c trng cho dao ng ca PO43- xut hin 466,27; 568,17-605,60; 963,30; 1100,55; 1065,20 cm-1 v vch dao ng ca lin kt O-H 3570,68 cm-1. Di hp th yu ca nc t do vng 3420,42 v 1645,91 cm-1. Hnh 3.15b l ph FTIR ca mu HA t vi. Cc vch c trng cho dao ng ca PO 43- xut hin 483,68; 530,63-637,57; 1031,43; 1099,25 cm-1 v vch dao ng ca lin kt O-H 3570,68 cm-1. C hai kt qu FTIR ny u ph hp vi cc d liu XRD v FTIR ca HA n pha v mu HA chun ca NIST.

3.3.3.2. nh SEM

24

Hnh 3.16: nh SEM ca HA t v trng Hnh 3.16 l nh SEM HA xp t v trng, cho thy sau phn ng thu nhit p sut cao, khung xp c cu to t cc tinh th HA dng ht, kch thc khng ng u v nm trong khong 50-100 nm. Cc ht khng lin kt vi nhau, gia chng tn ti cc bin r rt, chng t mu nhn c ch dng khi xp, cha to thnh gm HA.

Hnh 3.17: nh SEM ca HA xp t vi Mu HA sau phn ng thu nhit t vi p sut cao (hnh 3.17) gm cc tinh th HA dng ht, kch thc khng ng u v nm trong khong 100-200 nm. Cng tng t nh HA ch to t mai mc v san h [10,11,21,30,42], HA t v trng cng gm cc ht khng lin kt vi nhau, gia chng tn ti cc bin ht r rt, chng t mu nhn c ch dng khi xp, cha to thnh gm HA.

25

3.3.3.3. c trng TGA

Hnh 3.18: c trngTGA ca HA xp t v trng

Hnh 3.19: c trng TGA ca HA xp t vi Hnh 3.18 v 3.19 l c trng TGA ca HA xp t v trng v vi. Nh vy, c th khng nh HA tng hp c l n pha v bn nhit n 1000oC. 3.3.4. Mt s c trng ca gm xp hydroxyapatit 3.3.4.1. c trng XRD to thnh gm xp, cc mu HA xp tng hp t phn ng thu nhit c tip tc nung thiu kt nhit 1000oC trong 2 gi. 26

V-1000oC

Hnh 3.20: Gin XRD ca gm HA ch to t v trng, thiu kt 1000oC Trn gin (hnh 3.20) ch tn ti duy nht cc vch c trng ca HA, tinh th theo tnh ton tng ln, khong 95,7%. D-1000oC

Hnh 3.21: Gin XRD ca gm HA ch to t vi, thiu kt 1000 oC Trn gin (hnh 3.21) ch tn ti duy nht cc vch c trng ca HA, tinh th tng ln, khong 96,8%.

3.3.4.2. c trng SEM

27

Hnh 3.22: nh SEM ca HA t v trng sau khi thiu kt 1000oC Hnh 3.22 l nh SEM ca HA t v trng sau khi thiu kt 1000 oC cho thy phn ln bin ht gia cc tinh th HA bin mt v lin kt vi nhau to thnh gm xp HA. Cc l xp lin thng ba chiu vi nhau v c kch thc trong khong 200-500 nm.

Hnh 3.23: nh SEM ca HA t vi sau khi thiu kt 1000oC Hnh 3.23 l nh SEM ca ca HA xp t vi sau khi thiu kt. Kt qu nhn c cho thy cc ht HA lin kt vi nhau to thnh gm xp HA, bin gia cc ht HA khng cn tn ti. Cc l xp lin thng ba chiu vi nhau v c kch thc trong khong 100-300 nm.

3.3.4.3. c trng FTIR

28

Hnh 3.24: Ph FTIR ca gm HA t v trng Trn ph FTIR ca mu HA t v trng ch tn ti cc di hp th vng 1077; 1033; 963; 563-608 v 459 cm-1 c trng cho nhm PO43-, cc di 3570 v 635 cm-1 c trng cho nhm OH-. Di hp th ca H2O t do nm vng 3433 v 1638 cm-1 hu nh khng cn na.

Hnh 3.25: Ph FTIR ca gm HA t vi 29

Trn ph FTIR (hnh 3.25) ca mu HA t vi ch tn ti cc di hp th vng 1093; 1038; 962 v 561-608 cm-1 c trng cho nhm PO43-, cc di 3566 v 637 cm-1 c trng cho nhm OH-. Di hp th ca H2O t do nm vng 3433 v 1638 cm-1 cng khng cn tn ti. Gin XRD v ph FTIR ca mu gm xp HA sau khi thiu kt 1000oC v c bn ph hp vi c trng XRD v FTIR ca mu HA chun ca NIST [25].

KT LUN CHUNG 1. kho st qu trnh ch to HA bng phn ng thy nhit trc tip t CaCO3 dng canxit ca v trng v vi t nhin. Tuy vy, HA khng c to thnh hoc vi lng rt nh iu kin nhit 200 oC, p sut 14 atm trong thi gian 96 v 135 gi. 2. nghin cu tng hp c HA xp, n pha t v trng v vi bng phng php phn ng thy nhit thng qua giai on trung gian phn hy CaCO3 thnh CaO. S dng cc phng php vt l nhiu x tia X (XRD), hp th hng ngoi (FTIR), phn tch nhit (TGA) v hin vi in t qut (SEM) xc nh cht lng sn phm trn c s nh gi n pha, kch thc tinh th trung bnh, hnh dng v tinh th ca sn phm. 3. kho st nh hng ca cc yu t nhit , p sut v thi gian phn ng n n pha ca HA xp. Kt qu cho thy iu kin thch hp phn ng thu nhit to HA xp t v trng v vi l: nhit 200 oC,

30

p sut 14 atm, thi gian 24 gi. Gm xp HA c hnh thnh khi nung thiu kt HA xp 1000oC trong 2 gi. 4. Cc sn phm thu c u c cc c trng XRD, FTIR v TGA tng ng vi mu HA chun ca NIST. Cc nh SEM cho thy gm HA c cu trc xp v c to thnh t cc ht HA kch thc khong 100 -500 nm.

References TING VIT 1. inh Th Mng Cm, Nguyn Hu Ch, L Khc Bnh, Trn Tun, Nguyn Th Hi Yn, Trn Quang Trung (2007), Tng hp mng cng crm bng phng php phn x m ion (SIP), Tp ch pht trin KH&CN, 10(5). 2. V ng (2006), Cc phng php vt l trong ha hc, NXB i hc V Duy Hin, o Quc Hng, Phan Th Ngc Bch (2007), Tng hp v Quc gia H Ni. 3. kho st nh hng ca nhit n kch thc ht hydroxyapatit bng phng php kt ta ho hc, Tp ch Ho hc, 45(6A), tr.21-25. 4. V Duy Hin, o Quc Hng, Phan Th Ngc Bch (2008), Nghin cu ch to HA t khung xp t nhin ca mai mc bng phn ng thy nhit, Tp ch Ha hc, 46(2A), tr.118-123. 5. V Duy Hin (2010), Nghin cu ch to v c trng ha l ca hydroxyapatit dng gm xp c kh nng ng dng trong y sinh hc, Lun n Tin s Ha hc, Vin Ho hc - Vin Khoa hc v Cng ngh Vit Nam. 6. L V Tun Hng, Nguyn Vn n, Hunh Thnh t (2006), Nghin cu ch to mng mng TiO2 bng phng php phn x magnetron RF, Tp ch pht trin KH&CN, 9(6), tr.23-28. 7. o Quc Hng, Phan Th Ngc Bch (2007), Tng hp bt hydroxyapatit kch thc nano bng phng php kt ta ho hc, Tp ch Ho hc, 45(2), tr.147-151.

31

8.

Ngc Lin (2006), Nghin cu qui trnh tng hp bt v ch th gm xp

hydroxyapatit (HA), Bo co tng kt ti khoa hc cng ngh cp b, B KH&CN. 9. Ni. 10. Nguyn Tin Ti (1999), Nhit hc v phn tch nhit, Vin Ho hc, Vin Nguyn nh Triu (2006), Cc phng php vt l ng dng trong ha hc, Phan Vn Tng (2007), Cc phng php tng hp vt liu gm, NXB i KH&CN Vit Nam, H Ni. 11. NXB i hc Quc gia H Ni. 12. hc Quc gia H Ni. TING ANH 13. Alston Callahan, John Long (2001), Advances in Functional Oculoplastic Amit Y. Desai (2007), Fabrication and Characterization of Titanium-doped Andrew T. Metters, Chien Chi Lin (2007), Biodegradable Hydrogels: Surgery, Vision, 3(1). 14. Hydroxyapatite Thin Films, Journal of Materials Science., 30, p.679-684. 15. Tailoring Properties and Function through Chemistry and Structure, Taylor & Francis Group, LLC, Biomaterials, p.110-153. 16. Ariadne C.C.C., Mrcia T.P., Joslia D., Jos C.Z.S., Gibson L.P., Fbio A.S. (2006), Physico-chemical characterization and biocompatibility evaluation of hydroxyapatites, Journal of Oral Science, 48(4), p.219-226. 17. Billotte W.C. (2007), Ceramic Biomaterials, Taylor & Francis Group, Biltz R.M., Pellegrino E.D. (2007), The Composition of Recrystallized Celotti G., Landi E., La Torretta T.M.G., Sopyan I., Tampieri A. (2002), A Nguyn Hu Ph (2003), Ho l v Ho keo, Nh xut bn KH&KT, H

LLC, Biomaterials. 18. Bone Mineral, J. Dent. Res.,62(12), p.1190-1195. 19. novel sol-gel technique for hydroxyapatite preparation, Materials Chemistry and Physics, 78, p.816-824. 20. Certificate of Analysis (1997), Standard Reference Material 2910,

National Institute of Standards and Technology, Gaithersburg, MD, USA.

32

21.

Changsheng Liu, Yue Huang, Wei Shen, Jinghua Cui (2001), Kinetics of Chattopadhyay P., Pal S., Wahi A.K., Singh L., Verma A. (2007), Synthesis

hydroxyapatite precipitation at pH 10 to 11, Biomaterials, 22, p.301-306. 22.

of Crystalline Hydroxyapatite from Coral (Gergonacea sp) and Cytotoxicity Evaluation, Trends Biomater. Artif. Organs, 20(2), p.142-144. 23. Chen Y., Miao X. (2005), Thermal and chemical stability of fluorohydroxyapatite ceramics with different fluorine contents, Biomaterials (Singapore), 26(11), p.1205-1210. 24. Cuneyt Tas A. (2000), Combustion synthesis of calcium phosphate bioceramic powders, Journal of the European Ceramic Society, 20, p.2389-2394. 25. Donadel K., Laranjeira M.C.M., Goncalves V.L., Favere V. T. (2004), Structural, Vibrational and Mechanical Studies of Hydroxyapatite produced by wet-chemical methods, Depto de Qumica, Universidade Federal de Santa Catarina, Florianopolis, Santa Catarina, Brazil, Cx. P. 476, 88040-900. 26. Ferraz M.P., Monteiro F.J., Manuel C.M. (2004), Hydroxyapatite nanoparticles: A review of preparation methodologies, Journal of Applied Biomaterials & Biomechanics, 2, p.74-80. 27. Eliaz N., Sridhar T.M., Electronchemical and electrophoretic deposition of Eric J. Amis, John Rumble (2003), Calcium Hydroxyapatite, National hydroxyapatite for orthopaedic applications, Suface Engineering, 21(3). 28.

Institute of Standards & Technology, Certificate of Analysis Standard Reference Material 2910. 29. Gabrielle Murphy (2004), Hydroxyapatite Orbital Implants, ELE 482

Biomedical Engineering Seminar III, February 23rd. 30. Gmez-Morales J., Torrent-Burgus J., Rodriguez-Clemente R. (2001), Crystal Size Distribution of Hydroxyapatite Precipitated in a MSMPR Reactor, Cryst. Res. Technol., 36, p.1065-1074. 31. Gmez-Morales J., Torrent-Burgus J., Boix T., Fraile J., OdrguezClemente R. (2001), Precipitation of Stoichiometric Hydroxyapatite by a Continuous Method, Cryst. Res. Technol., 36, p.15-26.

33

32.

Guzmn Vzquez C., Pia Barba C., Munguia N. (2005), Stoichiometric

hydroxyapatite obtained by precipitation and sol gel processes, Revista Mexicana de Fisica, 51(3), p.284-293. 33. G. Verde-Carvallo, A. Guarino, G. Gonzlez (2004), Mineralization of Hydroxyapatite over Collagen Type I, European Cells and Materials, Vol.7. No.2, p.58-59. 34. Hien Vu Duy, Huong Dao Quoc, Bich Phan Thi Ngoc (2008), Study of the

Formation of Porous Hydroxyapatite Ceramics from Corals via Hydrothermal Process, International scientific conference Chemistry for development and integration, VietNam, p.1007-1013. 35. Hu J., Russell J., Vago R. and Ben-Nissan B. (2007), Production and analysis of hydroxyapatite from Australian corals via hydrothermal process,Journal of Materials Science Letters, 20(1), p.85-87. 36. Inagaki M., Yokogawa Y., Kameyama T. (2006), Highly Oriented Hydroxyapatite Coating Using rf Plasma Spraying, Key Engineering Materials, 309-311, p.615-618. 37. Ishizava H., Ogino M. (1996), Thin HA layer formed on porous titanium using electrochemical and hydrothermal reaction, J. of Mat. Sci., 31, p.6279 -6284. 38. Ivankovic H., Gallego Ferrer G., Tkalcec E., Ivankovic M. (2006), Preparation of highly porous hydroxyapatite ceramics from cuttlefish bone, Advances in Science and Technology, 49, p.142-147. 39. Jingbing Liu, Kunwei Li, Hao Wang, Mankang Zhu, Haiyan Xu, Hui Yan (2005), Self-assembly of hydroxyapatite nanostructures by microwave irradiation, Advanced Materials, 10(1), p.49-53. 40. Jung-Jae Kim, Hae-Jung Kim, Kang-Sik Lee (2008), Evaluation of Biocompatibility of Prous Hydroxyapatite Developed from Edible Cuttlefish Bone, Key Engineering Materials, 361-363, p.155-158. 41. Joon B. Park Joseph D. Bronzino (2007), Biomaterials - Principles and London New York Washington D.C., Applications, CRC Press Boca Raton p.18-38.

34

42.

Jyrki Vuola (2001), Natural coral and hydroxyapatite as bone substitutes,

Academic Dissertation, To be dicussed publicly with the permission of the Medical Faculty of the University of Helsinki, in the Faltin Room of the fourth Department of Surgery, Kasarmikatu 11-13, Jan. 5th. 43. Kothapalli C.R., Shaw M.T., Wei M. (2005), Biodegradable HA-PLA 3-D porous scaffolds: Effect of nano-sized filler content on scaffold properties, Acta Biomaterialia, p.653-662. 44. Kazue Yamagishi, Kazuo Onuma, Takashi Suzuki, Fumio Okada, Junji Tagami, Masayuki Otsuki, Pisol Senawangse (2005), A synthetic enamel for rapid tooth repair, Nature, 433, p.819. 45. Lawrence G. Brealt, Sung Y. Lee, Nicole E. Mitchell (2004), Fixed

Prosthetics with a Connective issue and Alloplastic Bone Graft Ridge Augmentation: A Case Report, The Journal of contemporary Dental Practice, 5(4). 46. Lu Xiaoying, Fan Yongbin, Gu Dachun, Cui Wei (2007), Preparation and Characterization of Natural Hydroxuapatite from Animal Hard Tissues, Key Engineering Materials, 342-343, p.213-216. 47. Luong Nguyen Dang, Nam Jae-Do, Synthesis of Hydroxyapatite and Application in Bone and Dental Regeneration in Human Body, Department of Polymer Science and Engineering; Sungkyunkwan University, Korea. 48. Masahiko T., Yoshihisa K., Kuniyoshi A., Yasuo S., Tsuyoshi K., Kiyoshi K., Akio M. (2003), Bone ingrowth fixation of artificial intervertebral disc consisting of bioceramic-coated three-dimensional fabric, J. Biomed. Mater. Res., Spine, 28, p.637-644. 49. Markovic M., Fowler B.O., Tung M.S. (2004), Preparation and comprehen characterization of a calcium hydroxyapatite reference materials, Journal of Research of the National Institute of Standards and Technology, 109, p.553-568. 50. Murugan R., Ramakrishna S. (2007), Development of Cell-Responsive Nanophase Hydroxyapatite for Tissue Engineering, American Journal of Biochemistry and Biotechnology, 3(3), p.118-124. 51. Narasaraju T.S.B., Phebe D.E. (1996), Some physico-chemical aspects of hydroxyapatite, Journal of Materials Science, 31, p.1-21.

35

52.

Nasser Y. Mostafa (2005), Characterization, thermal stability and sintering

of hydroxyapatite powders prepared by different routes, Materials Chemistry and Physic MAC 11004, 9. 53. Neih G. (2000), Synthesis and Charaterrization of porous Hydroxyapatite Orli S., Ivankovi H. and Tkalec E. (2005), Preparation of highly porous and Hydroxyapatite coating, Lawren Livermore Lab., USA, p.4-13. 54. hydroxyapatite ceramics from cuttlefish bone, University of Zagreb, Faculty of Chemical Engineering and Technology, Zagreb, Croatia. 55. Pekka Ylinen (2006), Applications of coralline hydroxyapatite with bioabsorbable containment and reinforcement as bone graft substitute, Academic Dissertation, University of Helsinki. 56. Peron E., Fuentes G., Delgado J.A., Morenjon L. Almirall A., Garca R. (2004), Preparation and characterization of porous blocks of synthetic hydroxyapatite, Latin American Applied Research, 34, p.225-228. 57. Phebe D.E. (1996), Some physico-chemical aspects of hydroxyapatite, Pierre Layrolle, Atsuo Ito, Tetsuya Tateishi (1998), Sol-Gel Synthesis of

Journal of Materials Science, 31, p.1-21. 58.

Amorphous Calcium Phosphate and Sintering into Microporous Hydroxyapatite Bioceramics, Journal of the American Ceramic Society, 81(6), p.1421-1428. 59. Qiyi Z., Yang L. (2005), Electrodchemical activation of titanium for biomimetic coating of calcium phosphate, Biomaterials, 26, p.3853-3859. 60. Raynaud S., Champion E., Bernache-Assollant D., Thomas P. (2002), Calcium phosphate apatites with variable Ca/P atomic ratio I. Synthesis, characterisation and thermal stability of powders, Biomaterials, 23, p.1065-1072. 61. Robert E. Dinnebier, Karen Friese, (1999), Modern XRD methods in mineralogy, Max-Planck-Institute for Solid State Research, Stuttgart, FRG. 62. Rocha H.G., Lemos A.F., Kannan S., Agathopoulos S., Ferreira J.M.F. (2005), Hydroxyapatite scaffolds hydrothermally grown from aragonitic cuttlefish bones, J. Mater. Chem., 15, p.5007-5011.

36

63.

Satou K. (2001), Rhemeimetric Characteristics of slurry coutrolling the and the compressive strength behavior of biomimetic

microstructure

hydroxyapatite, Materials Research Soc., 16(1), p.164-165. 64. Shin H., Shinsuke I., Jiro T., Taizo F., Masashi N., Yoshitaka M., Yasuo S., Masaki O., Takashi N. (2007), A 5-7 year in vivo study of high-strength hydroxyapatite/poly(L-lactide) composite rods for the internal fixation of bone fractures, Biomaterials, 27, p.1327-1332. 65. Shin H., Jiro T., Masashi N., Koji G., Yasuo S., Makoto S., Masakazu K., Takashi N. (2005), In vivo evaluation of a porous hydroxyapatite/poly-DL-lactide composite for use as a bone substitute, J. Biomed. Mater. Res. A.,75, p.567 -579. 66. Shinsuke I., Jiro T., Taizo F., Takashi N., Yoshitaka M., Yasuo S., Masaki O. (2005), Long-term study of high-strength hydroxyapatite/poly (L-lactide) composite rods for the internal fixation of bone fractures: a 2-4-year follow-up study in rabbits, J. Biomed. Mater. Res., 66B, p.539-547. 67. Shikhanzadeh M. (1991), Bioactive Calcium photphate Coating Prepared by Electrodeposition, J. of Mat. Sci. Let., 10, p.1415-1417. 68. Tin-O.M.M., Gopalakrishnan V., Samsuddin A.R., Al Salihi K.A., Shamsuria O. (2007), Antibacterial property of locally produced hydroxyapatite, Archives of Orofacial Sciences, 2, p.41-44. 69. Van Houtte P., Vander Sloten J., Van der Perre G., Van Audekercke R. (2005), Porous scaffolds for the replacement of large bone defects: a biomechanical design study, Celestijnenlaan 300-B-3001 Heverlee (Leuven), Belgium. 70. Verde-Carvallo G., Guarino A., Gonzlez G. (2004), Mineralization of Willi Paul and Chandra P. Sharma (2001), Porous Hydroxyapatite Hydroxyapatite over Collagen Type I, European Cells and Materials, 7(2), p.58 -59. 71. Nanoparticles for Intestinal Delivery of Insulin, Trends in Biomaterials & Artificial Organs, 14(2), p.37-38. 72. Xing Zhang and Kenneth S. Vecchio (2007), Hydrothermal synthesis of hydroxyapatite rods, Journal of Crystal Growth, 308(1), p.133-140.

37

73.

Young R.A., Mackie P.E. (1980), Crystallography of human tooth enamel:

Initial structure refinement, Mat. Res. Bull., p.17-29. 74. Yu Ji Yin, Feng Zhao, Xue Feng Song, Kang De Yao, William W. Lu, Leong J. (2001), Preparation and characterization of Chiyan

hydroxyapatite/chitosan-gelatin network composite, Journal of Applied Polymer Science, 77(13), p.2929-2938.

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