Numerical Simulation for Distribution and Mass Transfer of Nutrient Substance within Microbeads in Different Culture Systems

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a n i h C , 4 2 0 6 1 1 n a il a D , y g o l o n h c e T f o y ti s r e v i n U n a il a D

2_B_u_r_n_s_R_e_s_e_a_r_c_h_G_r_o_u_p_,_A_N_Z_A_C_R_e_s_e_a_r_c_h_I_n_s_t_ti_u_t_e_,_U_n_i_v_e_r_s_ti_y_o_f_S_y_d_n_e_y_,

a il a r t s u A , 9 3 1 2 , W S N , d r o c n o C

r o h t u a g n i d n o p s e r r o C *

s d r o w y e

K :Numerica ls uim laiton ,Masstransfer,Microbeads ,Cutluresystem.

.t c a r t s b

A Bioreactori sagooddeviceforbiologica landchemica lreactions.Microbeadshavegood .

y ti l i b a e m r e p d o o g r i e h t f o e s u a c e b s d l e i f y n a m n i e u l a v n o it a c i l p p

a This research was aimed to

t e t a l u m i

s hemasstransferprocessofglucoseviamicrobeadsindifferen tculturesystemsincluding T-flasks ,rotating cel lculture systemsa nnd ovel rotary perfusion bioreactor with Fluen tsoftware .

s w o l l o f s a e r a s n o i ti d n o c l a it i n i e h

T . Themassfraction of glucoseinsideand outside thebal lwas 0 0. da n 4.5e-6_._T_h_e_q_u_a_il_t_y_s_o_u_r_c_e_w_a_s₆_.₀₅_e-6_,_a_n_d_t_h_e_t_i_m_e_s_t_e_p_w_a_s₁_e-3_._W_h_e_n _t_h_e_T_-_f_l_a_s_k_w_a_s_a_t

n r u t h t 0 1 e h

t da n the itme was a t45s ,the concentraiton of glucose insideand outside the sphere q

e h c a e r y l l a c i s a b d l u o

w uilibrium. Fortheglucoseconcentration ti wouldreachequilibrium a tthe n

r u t h t 0

1 after30s . tI wasconfirmed tha tthemasstransfereffec tw asbetterinRotary cel lculture m

e t s y

s thant hati nothersystems.

Introduciton

p i u q e f o d n i k a e r a s r o t c a e r o i

B men twhich areused forbiologica lorbiochemica lreactions ,and n

a

c also monitor and contro limportan tparameters in culture condiitons such as biomechanics , ,

e r u s s e r

p pH ,temperature ,metabolism of nutrients and wastes and mass transfer as wel las 3D l

u

c tivation[ - ]13 _.₃_D _e_n_v_i_r_o_n_m_e_n_t_w_h_i_c_h_i_s_m_a_x_i_m_u_m_c_l_o_s_e_t_o_t_h_e_n_a_t_u_r_a_l_e_x_t_r_a_c_e_l_l_u_l_a_r_m_a_t_r_i_x_c_a_n_b_e

l l e c f o s e p y t c i f i c e p s e r u t l u c n a c e w t a h t o s , r o t c a e r o i b n o g n i y l e r d e t a e r

c s in vtiro [4]_._C_a_l_c_i_u_m

o r c i m e t a n i g l

a beads ,as a carrier of immobilized cel ltechnology ,are commonly used for cel l d

e n i a t s u s g u r d d n a n o it c e t o r p l l e c , n o i t a v i t l u

c -release. I t has many properti es such as good

ti l i b a e m r e

p y ,no obvious toxictiy ,metabolitebiocompatible ,simpleoperation and easily breaking l

l a w e h

t ,smal ldiffusionresistanceofnutrien tsubstance ,mildconditionsforimmobiilzationandso n

o ,somecellswhicharefragileandverysenstiivetofluidshearstress ,mus tbeembeddedin micro e

g a m a d m o r f t c e t o r p o t s d a e

b [ - ]59_._T_h_e_r_e_s_e_a_r_c_h_o_n _n_u_t_r_i_e_n_t_d_i_s_t_r_i_b_u_t_i_o_n _o_f_d_i_f_f_e_r_e_n_t_t_y_p_e_s_o_f_m_i_c_r_o

s s a m d n a t s r e d n u s u e k a m n a c s r o t c a e r o i b n i s d a e

b characteristicsandprinciples oftransferinmicro r

e h t e h w e r o l p x e d n a , s d a e

b encapsulated micro beads wil laffec tthe absorption of nutrients of l

a n r e t n

i cells [ - ]1011 _._S_i_m_u_l_t_a_n_e_o_u_s_l_y_,_i_t_c_a_n _a_l_s_o _p_r_o_v_i_d_e _r_e_f_e_r_e_n_c_e _d_a_t_a _f_o_r _t_h_e _o_p_it_m_i_z_a_it_o_n _o_f

s r o t c a e r o i

b withcultureconditions. h

T is study eu d s fluent software to simulatetheglucose masstransferprocess of microbeads in T-flask ,rotary cel lcutlure system as wel las nove lrotary perfused bioreactor ,respectively .The

t n e m i r e p x

e focused on numerica lanalysis for differen tflow fie s ld inside different bioreactors. tI e

d i v o r

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Methods t n e u l

F So tfwareMathemati Mc s o del

w o l f d i u l f e t a l u c l a c o t d e s u s a w ) A S U , A P , g r u b s n o n a C , S Y S N A ( e r a w t f o s t n e u l F S Y S N A

d i r g l a e d i s e d i v o r p t I . m e t s y s c i r t e m o e g x e l p m o c a n i r e f s n a r t s s a m d n a t a e h d n a , n o i t u b i r t s i d

e r r i n a y b y l i s a e s m e l b o r p d i u l f x e l p m o c e v l o s n a c d n a y t il i b a t p a d

a gulargrid ,whichisprovidedby

. y r t e m o e g x e l p m o c

g n it t e

S Condi iton so fMulitphaseSimula itonReactors e

h

T Phy isca lMode lo fReactors .As shown in Figure 1 ,theseare diagrams o T-f flask ,rotary cel lcutluresystemdevicesandnove lrotaryperfusedbioreactor ,respectively.

g i

F u 1. )re ( T-A flask ;(B)Rotarycel lculturesystem ;(C)Nove lrotaryperfusedbioreactor.

l a it a p S ( g n i d d i r

G Dsicreitza iton). T-flask :Compared to the size of T-flask , et h radius of s

d a e b o r c i

m w asso smal ltha tthebeadsbecamea point .Masstransferon thesurfaceofthebeads a

h d asignifican timpac tonly on theflowfield nearthebeads .Iftheborderisrelatively small ,the l

e d o

m can be simplified into a two-dimensiona l plane . Parameters of the simpilfied o

w

t -dimensiona lmode loft hemicrobeadsandrelativeboundarycondiitonswereshowni nTable1. m

e t s y s e r u tl u c l l e c y r a t o

R (RCCS) :Thissystem can also besimplified into a two-dimensiona l e

n a l

p a s the reason with above menitoned T-flask model . Parameters of the simplified o

w

t -dimensiona lsimulaiton mode lof the microbeads and relaitve boundary conditions werealso s

d a e b o r c i m r u o F . 1 e l b a T n i n w o h

s were se tinto the culture chamber ,and they were relatively e

h t h t i w c i t a t

s bioreactorwheni twasrotating ,3Dmode loft hissystemw asshowni nFigure.2 . r

o t c a e r o i b d e s u f r e p y r a t o r l e v o

N (NRPB) :T henumberofmicrobeadsshowed an effects on the y

c n e i c i f f e r e f s n a r t s s a

m of this nove lbioreactor. And the 3D grid mode lof NRPB w asshown in e

r u g i

F 2 ,four or five micro beads were arranged in a row into the culture chamber .During the y

e h t , n o i t a t o

r werek te p relativelystaticwithNRPB. o

w t d e i f i l p m i s e h t f o s r e t e m a r a P . 1 e l b a

T -dimensiona lmode loft hemicrobeadsandBoundaryconditions.

r e d r o

B raBdailu ls ) m m (

e b o l G

e c n a t s i d

) m m

( Mode lstate

s n o i t i d n o c l a i t i n I

e s o c u l G

s s a m

n o i t c a r f

i/o e r e h p s

Quality e h t f o

e c r u o s

m r e t

e m i T

p e t s

T-flask 30x30mm2 ₁_.₁ ₁₀ _S_t_a_t_i_o_n_a_r_y ₀_.₀_/₄_.₅_e-6 ₆_.₀₅_e-6 ₁_e-3

S C C

R Radiusi s30mm 1.1 1 0 Diskrotationa lspeedi s20rpm 0.0/4.5e-6 ₆_.₀₅_e-6 ₁_e-3

P R

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s n o it i d n o

C S te it .ng (1)Microbeadswiththesamediameterhad thesamei nterna lporestructure. e

h t e d i s t u o n o it a r t n e c n o c n i a m e h T ) 2

( microbeadsw asuniform.

) 3

( Microbeadsw iththesamediameterha dthesamei nterna ldiffusioncoefficien.t )

4

( Soluteconcentraitondistributionw asonlyalongt heradia ldirecitonofmicrobeads.

Resutls T-lfask

e h

T distributionofmassfractionofglucoseo nthesurfaceofmicrobeads Tin -flaskswereshowni n e

r u g i

F .3&4 ,i twasfound tha ttheglucoseconcentration insidebeadsgradually increased wtih time i tn h e mass transfer progress of microbeads, while the concentration outside decreased slightly.

T e h t n e h

W -flask was a tthe 10th turn after 45s ,the concentration of glucose inside and outside s

d a e b o r c i

m would basically reach an equliibrium .There w as a very significan tconcentraiton m

o r f n o i t u b i r t s i d t n e i d a r

g eachoffourcornerst owardt hecenter .Thet hicknessofsurfacemasswas m

u m i x a

m in thecenterofthemicrobeads ,and thethicknessinsidebeadsw asgreaterthan tha tof .

e d i s t u o e h

t I twasproved tha tglucose molecules could be driven uniformly into the microbeads .t

n e i d a r g n o it a r t n e c n o c e h t y b r o i r e t n i

g i

F u 3. re Distributionofmassfractionofglucosei nt hesurfaceofsphere. (A)15s ;(B)33s ;(C)36s ;(D)39s ;(E)45s.

g i

F u 4. re Distributionofmassfractionofglucosei nt hesurfaceofsphere. (A)42s ;(B)4 . 5s

y r a t o

R C lle Cutlure System

d e h

T istribution ofmassfraciton ofglucoseo n thesurface ofmicrobeadsin RCCS wasshown in e

r u g i

F 5 .I twasfoundthattheglucoseconcentrationi nsidet hemicrobeadsi ncreasedgraduallywith e

m i

t while the outside concentration showed a slightly decreasing trend . The closer to the w

n o i t a r t n e c n o c e h t r e w o l e h t , s d a e b o r c i

m ould beand thelowes tconcentration w aslocated in the s

a e r c e d e h t d n a s d a e b o r c i m r u o f f o n o i t i s o p l a r t n e

c ingtrendchanged relatively uniform .Whenthe

n r u t h t 0 1 e h t t a s a w r o t c a e

r after30s,t heglucoseconcentrationinsideandoutsidet hespherewould t

I . m u i r b i l i u q e n a h c a e

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F gi u 5. re Distributionofmassfractionofglucosei nt hesurfaceofsphere .(A)0.5s ,1/6cycle ;(B)18s ,6cycle ;(C)21s , e

l c y c

7 ;(D)30s ,10cycle.

l e v o

N RotaryPerfusedBioreactor

T he distribution of mass fraciton of glucose outside and inside the microbeads in a NRPB w as w

o h

s n in Figure.6&7 .As shown in Figure.6 ,the concentration of glucose inside the microbeads y

l l a u d a r g d e s a e r c n

i withtimewhlietheconcentraiton ofglucoseoutsidethemicrobeadsdecreased h

t 5 e h t t a s a w r o t c a e r e h t n e h W . y l t h g i l

s turn after 30s ,the concentration of glucose inside and s

d a e b o r c i m e h t e d i s t u

o would reach an equilibrium. Figure 6A&B showed tha t glucose e

h t e d i s t u o n o i t a r t n e c n o

c beads showed a decreasing trend, the partia lconcentration reached a e

u l a v m u m i n i

m in thecentra lportion. tI wasproved tha tthemasseffects w asbetterin theNRPB T

n i t a h t n a h

t -flasks. The concentration of glucose in thetwo microbeads on theoutside was the o

w t e h t d n a , t s e w o

l in themiddlewasslightly highe.r However ,thegap ofthe concentration w as n

o i t a r t n e c n o c r e w o l y l e v i t a l e r e h t f o e s u a c e B . t a e r g t o

n i n the centra lportion ,the concentration

t n e i d a r

g w aslarger .Theeffec tofrotationa lspeed w asno tobvious in thetwo simulation process . 2

1 f o s n o i t i d n o c r e d n u , r e v e w o

H rpm ,theg ul cosemassfraciton ofsurfacemicrobeadsw ashigher . ,

d r o w r e h t o n a n

I due to high rotation speed ,the microspheremasstransfer efficiency washigher , t

d n

a hentheequliibriumwasreachedfaster .Presumablybecausetherotaitonspeedw aslarge ,fluid a

e h

s rstressi ncreased ,sot hatt heprocessofglucosemoleculesi ntomicrobeadsw asaccelerated.

g i

F u 6. re Distributionofmassfractionofglucoseoutsideandi nsidet hesphere .(A)4microspheres ,10rpm,40s ,4cycle; )

B

( 4microspheres, 12rpm ,40s ,3.3cycle; )(C 5 microspheres, 10rpm ,18s ,3cycle; )

D

( 5 microspheres, 10rpm ,30s ,5cycle. n

i n w o h s s

A Figure7 ,the concentration of glucose insidethe micro beads increased gradually e

m i t h t i

w whiletheconcentraiton of glucoseoutside themicrobeadsdecreased slightly .When the n

r u t h t 6 e h t t a s a w r o t c a e

r after 30s ,the concentration of glucose inside and outside the sphere a

h d n a m u i r b i l i u q e n a h c a e r d l u o

w d atrend tostabilize.Thenumberofmicroplasticbeadsrotating r

t s s a m f o y c n e i c i f f e n o t c a p m i s a

h ansferin micro plasticbeads .Fivemicro plasticbeadswerese t .

e n i l a s a r e b m a h c r o t c a e r e h t n

i During the rotation ,the five ge lbeads remaine ld reaitvely n

e h w y r a n o i t a t

s rotated in theperfusion bioreacto.rFrom the simulation results ,i twasshown that thedistributionofglucoseconcentraitonchangesregularlyatt hebeginning, tb u i thadat endencyt o

. e m it f o d o i r e p t r o h s y r e v a n i e g r e v n o

c tI w as proved tha tthe materia lcould be uniformly n

i d e t u b i r t s i

(5)

g i

F u 7. re Distributionofmassfractionofglucoseoutsideandi nsidet hesphere 5( microspheres) .(A)12rpm ,12s ,2.4 e

l c y

c ;(B)12rpm ,15s ,3cycle ;(C)12rpm ,30s ,6cycle.

Dsicus ison C& onclu isons

r t s i d e h

T ibutionofnutrientsindynamicbioreactorwasmoreuniform ,which avoidedtheinfluence f

o low nutrients in the loca larea owing to the cel labsorpiton within the microbeads. As the t

a r t n e c n o

c ion of glucose around the beads aw s reduced, the concentraiton gradien twould n eot b n

e k a e

w ed ,which ensured a higher masstransferrate. tI was proved tha tthemasstransferrate of s

d a e b o r c i

m w asfasterinadynamicenvironmentcomparedt ostaitccultureenvironment . l

f e h

T owstatusin culturechamber fo NRPBw a as comparativelystandard laminarflow ,andthe s

d a e b o r c i

m h d a almostt hesameshearstressa tthesamepostiion ,soconcentraitongradien twast he r

o t c a f y l n

o which influenced mass transfer , which could explain the phenomenon shown in e

r u g i

F .6D. Figure 7 rp o d ve tha tmateria lcould be uniformly distributed in NRPB ,and theiniita l c

e c n e r e f f i d n o i t a r t n e c n o

c ouldbeneglected.Thevariationt rendofconcentrationdistribuitonshown 7

e r u g i F n

i C aw sdifferentfromtha toffourmicrobeads ,whichperhapsresulte d from thefactorof f

o n o i t u b i r t s i d t i x

e chamber ,perfused materia lfluid aligne d exactly with the middle microbeads , n

e h t d n

a thedistributionofglucoseconcentrationwasi nconsistentinfivemicrobeads. 0

1 ( s n o i t a l u m i s s s e c o r p o w t e h t g n i z y l a n a y

B rpmand 12rpm) ,i twasdeduced tha tthetrendsin s

e s s e c o r p n o it a l u m i s o w t e h

t were accordant. Although the difference of rotaitn g speed was no t ,

s u o i v b

o theglucosemassfractionofsurfacemicrobeadswashigherunderconditionsof12rpm .In g

i h , d r o w r e h t o n

a h rotaitn g speed mad e grea tmass transfer efficiency and was easy to reach an e

h t y l b a m u s e r P . m u i r b i l i u q

e rotatin g speed was large and fluid shear stress increased ,so tha tthe s

d a e b o r c i m o t n i s e l u c e l o m e s o c u l g f o s s e c o r

p w asthereforeaccelerated.

l w o n k c

A edgments

a n i h C f o n o i t a d n u o F e c n e i c S l a r u t a N l a n o i t a N y b d e t r o p p u s s a w k r o w s i h T

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[ A. Bakir , .J. S Rowland , .C. R Thompson .Enhanced desorption ofpersisten torganicpollutants .

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