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1 488,, 1, gm l, gl 2 (CPPfgPLLA ) 40%, 72 4, 18 : A, ; B CPPfgPLLA ; C ; D X A 2, ; 4, ; 8, ; 16,, 43. 6% B C D 3 2 ; 4, ; 16, ; 2,, REPA IR OF UPPER TIB IAL EP IPHY SEAL D EFECT W ITH ENGINEERED EP IPHY SEAL CARTILAGE IN RAB2 B ITSgZH OU Q iang, L I Q i2hong, DA I Gang, et al. D ep artm ent of O rthop aed ics, S ou thw est H osp ital, T h ird M ilitary M ed ical U niversity. Chong qing, P. R. Ch ina Abstract Objective To observe the effect of engineered ep iphyseal cartilage regenerated in vitro w ith 32D scaffo ld by chondrocytes from ep iphyseal p late in repairing the tibial ep iphyseal defect, and to exp lo re the m ethods to p romo te the confluence betw een engineered cartilage and ep iphyseal p late. M ethods Chondrocytes w ere iso lated enzy2 m atically from the ep iphyseal p lates of imm ature rabbits, and then p lanted into the tissue culture flask s and cultivated. T he first passage chondrocytes w ere co llected and m ixed fully w ith the self2m ade liquid bio logical gel at app roxim ately cellsgm l to fo rm cell2gel fluid. T he cell2gel fluid w as dropped into the po rous calcium po lypho sphate fibergpo ly2 L 2lactic acid (CPPfgPLLA ) scaffo ld, and a cell2gel2scaffo ld comp lex fo rm ed after being so lidified. T he defect models of 40% upper tibial ep iphyseal p late w ere m ade in 72 imm ature rabbits; they w ere divided into 4 group s: group A (the cell2 gel2scaffo ld comp lex w as transp lanted into the defect and the gap filled w ith chondrocyte2gel fluid), group B (w ith non2 cell CPPfgPLLA scaffo ld), group C (w ith fat) and group D (w ith no th ing). T he changes of roen tgenograph, gro ss and h isto logy w ere investigated after 2, 4, 6, 8, 12 and 16 w eek s of operation. Results In group A, the typ ical h isto logi2 cal structure of ep iphyseal p late derived from the engineered cartilage w ith a fine integration betw een ho st and dono r tis2 sues after 2 w eek ṡ T he repaired ep iphyseal p late had no rm al h isto logical structure w ithout defo rm ation of tibia after 4 w eek ṡ T he early h isto logical change of ep iphyseal clo sure appeared in the repaired area w ith varus and sho rtening defo r2 m ation of the tibia after 8 w eek ṡ T he ep iphyseal p late w as clo sed in the repaired area w ith mo re evident defo rm ation of tibia; the grow th function of repaired ep iphyseal p late w as 43. 6% of the no rm al one. In group s B, C and D, defo rm a2 tion of tibia occurred after 2 w eek s; the defect area of ep iphyseal p late w as comp letely clo sed after 4 w eek ṡ T he defo r2 m ation w as very severe w ithout grow th of the injured ep iphyseal p late after 16 w eek s, and no significant difference w as observed betw een the th ree group ṡ Conclusion Engineered ep iphyseal cartilage can repair the ep iphyseal defect in the h isto logical structure w ith partial recovery of the ep iphyseal grow th capability. Injecting the suspension of fluid chondro2 cyte2gel into the defects induces a fine integration of ho st and dono r tissueṡ Key words T issue engineering ep iphyseal p late cartilage Chondrocytes R epair Founda tion item: N ational N atu ral Science Foundation of Ch ina ( ) [ 1 2 ], [ 2 5, ] : ( ) : (, ),,, China Academic Journal Electronic Publishing House. All rights reserved.
2 CPPfgPLLA ; C (Sigm a) 3 :, ; D [ 3 ],, gcm 2 10% (H yclone) 584 m ggl L ( ) 0. 1 mm o lgl (Gibco) 0. 4 mm o lgl ( ) 50 ΛggL C (Sigm a) DM EM gh am s2f12 (H yclone), 175 cm 2 (N unclon ), % CO 2,, 90% X, (Sigm a) 1 X 16 ( ) 1. 2 ( ) [ 6 ], gm l 1 :, (cal2 ( ) = - cium po lypho sphate fiber, CPPf) gl 2 (po ly2l 2 lactic acid, PLLA ) ( 11 mm, 2 mm, > 92% ), 6 (% ) =,, 37 5% CO 2 (mm ) - 15 (mm ), 5,, 10 (mm ) - 15 (mm ) 100% , ;, H E, gcm cm 2, 90%, gm l,, 2 M icro soft Excel 2000 t 1. 4, P < ( ), g, 2 m lgkg 1. 5% 2. 1 X ( ) A : 2 4, 115 mm,,, 014 mm 6 ; 8, 15 mm, 9 5, ( 1) ; 10 12,, ;,, 1 mm 16 ( 2),,, 40%, B C D 3, 1. 5, ; B : :,,, 4% 15% ED TA 22N a, (% ) = (mm ) - (mm ) 100% (mm ) ; 2 ; 4 72,, ; 16 4, 18 A : ( 3), China Academic Journal Electronic Publishing House. All rights reserved.
3 490 1 A 8 X, 2 A 16 X, 3 D 16 X, F ig. 1 Roen tgenogram of group A in the 8th week, light varus and shorten ing deformation can be seen in the r ight tibia ( ) F ig. 2 Roen tgenogram of group A in the 16th week, the deformation of r ight tibia get more ev iden t ( ) F ig. 3 Roen tgenogram of group D in the 16th week, severe deformation can be seen in the r ight upper tibia ( ) 4 6, ( 5) ; 8, 16 X B C D 3, 2, : A, 4 3 (P < 0. 01),, 3 (P < 0. 01) ; B C D 3 (P > 0. 05), A 3 ; B C D,, 3, 2,, ( 6) ; 4,, ; 8, 16 X (n= 3, θ x s) Tab. Data of the r ight tibia on roen tgenogram s in the 16th week of each group (n= 3, x θ s) Group (% ) ( ) (% ) Percent of sho rtening A ngle of varus ( ) Percent of grow th recover A Group A B Group B C Group C D Group D B C D 3 P < Compared w ith group s B, C and D, P < ; 12 ; A : 2,,,, ( ) ( 7) ; 12 16,,,,,, B C D 2, B CPPfgPLLA, A : 2 C, D, ( 4) ; B ; China Academic Journal Electronic Publishing House. All rights reserved.
4 A 2, 5 A 4, 6 A 2, ( ) (Ρ ),,, ( 40) 7 A 8, ( ), (Ρ ) (HE 40) 8 D 4, (HE 40) F ig. 4 Repa ir of epiphyseal plate defects( ) of group A in the 2nd week F ig. 5 Repa ir of epiphyseal plate defects( ) of group A in the 4th week F ig. 6 In the 2nd week, the reparative epiphyseal plate ( ) wasmarkedly wider than the normal (Ρ ) of group A with clear boundar ies and without gap between them (Toluidine blue 40) F ig. 7 The formation of a bone plate ( ) under the reparative epiphyseal plate and a marg inal bone br idge (Ρ ) in the 8th week of group A (HE 40) F ig. 8 The exten sive bone formation ( ) in the epiphyseal plate defect of group D in the 4th week (HE 40) ( 8),,, [ 8 ] [ , ] L angen sk i o ld (1967),,,,, Jouve, [ 4 ] 30% ( ), 21 [ ] L ee : [ 3 ], China Academic Journal Electronic Publishing House. All rights reserved.
5 492,,, [ 5, ],,, [ 3 ],,,,,,,,, CPPfgPLLA, 40%,, %, 3 ; 2,,,,,,, X, ; 5,,,,,, 8,,,,,,,, Reconstr M icro surg, 1998; 14 (2) : 137,,,., 2001; 8 (10) : 980, Cap lan 1,.., 2001; 15 (5) : 291 2,.., 2003; 25 (2) : L ee EH, Chen F, Chan J, et al. T reatm ent of grow th arrest by transfer of cultured chondrocytes into physeal defects. O rthop, 1998; 18 (2) : 155 [ 14 ] J Pediatr 4 Jouve JL, Co ttalo rda J,M o ttet V, et al. Reimp lantation of grow th p late chondrocyte cultures in central grow th p late defects: Part g. Surgical experim entation in rabbits. J Pediatr O rthop B, 1998; 7 (2) : 174.., 2001; 23 (10) : ,,,.., 2002; 22 (6) : (PBR )., 1996; 16 (6) : O sterm an K. H ealing of large surgical defect of the ep iphyseal p late. A n experim ental study. C lin O rthop, 1994; 300: Glickm an AM, Yang JP, Stevens D G, et al. Ep iphyseal p late transp lantation betw een Pediatr O rthop, 2000; 20 (3) : 289 sites of different grow th po tential. J 10 Innocenti M, Ceruso M, M anfrini M, et al. F ree vascularized grow th2p late transfer after bone tumo r resection in children. 11 V ilkki SK. D istraction and m icrovascular ep iphysis tranfer fo r radial club hand. J H and Surg (B r), 1998; 23 (4) : ,,.., 2001; 15 (1) : 53 13,,,. 14 Cap lan A I, E lyaderani M, M och izuk i Y, et al. P rincip les of carti2 lage repair and regeneration. C lin O rthop, 1997; 342: 254 ( : : ) J China Academic Journal Electronic Publishing House. All rights reserved.
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