Preparation of compound tissue2engineering scaff olds of PLA/ silk f ibroin and evaluation of its biological features

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1 ( ) Journal of Jilin University ( Medicine Edition) Vol. 34 No. 3 May (2008) /,,, (, ) [ ] : ( PLA) / ( PL A/ ), : PLA, PLA/ PLA ( ) PLA/ ISO 10993,, :, ; < 012, < 110, ; < 5 % ( P = 01000), PLA/ PLA ;, PLA/ (37 min) PLA (26 min), PLA/ PLA ;, PLA/ PLA, PLA/ PLA : PLA/ PL A, [ ] PLA/ ; ; [ ] R318 [ ] A Preparation of compound tissue2engineering scaff olds of PLA/ silk f ibroin and evaluation of its biological features XIN G Shuai, XIA Ya2yi, YUAN Ling2wei, L U Mao2shen (Department of Orthopedics, Second Hospital, Lanzhou University, Lanzhou China) Abstract : Objective To study the preparation method of compound tissue2engineering scaffolds of the PLA/ silk fibroin and evaluate its biological features. Methods The PLA scaffolds matrix were dipped into the silk fibroin solution, then dried, and PL A/ silk fibroin scaffolds were prepared. There were two groups in the experiment, one group was PL A group, and the other one was compound scaffolds group. According to ISO standard, hematolysis test, dynamic coagulation time test, cell toxicity test, stimulation test and pyrogen test were performed in two groups, and the results were compared betwen two groups. Re sults In the stimulation test, the two kinds of materials had equally not aroused the obvious animal skin stimulation, it showed that the experiment was in accordance with the standard. In the pyrogen test, the two scaffolds material aroused the animal temperature rising without exception under 01 2 and the total number of degree was under 11 0, therefore there was no obvious difference between two groups. In the hematolysis test, the hemolysis rates of the two scaffolds samples were smaller than 5 % equally ( P = 01000), which indicated that the hemolysis of the compound scaffolds was better than that of the PL A scaffolds. In the dynamic coagulation time test, the coagulation time of the compound (ZS0112A Y) ( ),,,, ( Tel : , E2mail :

2 406 ( ) scaffolds ( 37 min) was longer than that of the PL A scaffolds ( 26 min). The anti2coagulation ability of the compound scaffolds was better than that of the PL A scaffdds. In the cell toxicity test, the cell growth situation of the compound scaffolds group was obviously better than that of the PLA group, and at the meantime the cell toxicity of the compound scaffolds was obviously smaller than that of the PLA scaffolds. Conclusion The material of PLA/ silk fibroin compound scaffolds has the advanced biological consistent compared with the simplex scaffolds. Accordingly, the PLA/ silk fibroin can be used as a scaffolds matrix to be transplanted into the body. Key words : PL A/ silk fibroin ; biological evaluation ; tissue engineering,,, [1 ],, (polylactic acid, PL A) [ 2 ] PL A, PL A, [3 ] PL A PL A, [4 ] PL A/, ; 112 PL A (, m, 90 %) ; ( ) ; ( ) ; DM EM/ F12 ( Gibco) ; (Sigma) ; ( ) ; ( TN [5 ] PL A, ), CO2, ; Lazzeri [6 ] / PLL A, PL A ; (B H22 Olymp us, ) ; 113 PL A/ :, (CaCl2 ), 40 % CaCl2,, 015g (CaCl2 ) (, [7 ] 60 min, Unger [8 ],,, ;, PL A, PL A/, ISO 10993, PL A/ ( PL A/ ), , kg, (,, : , : ) CaCl2 1 10),, CaCl2, CaCl2, 72 h, [9 ] : PLA 72 h,,, PL A/ 60 Co 114 PL A PL A/ 011 cm 015 cm, 0156 cm 2 ISO

3 , 1 / 407, / = 3 cm 2 L - 1,, h, g PLA, 3 % 5 %, PL A/,, kg 5 mm mm, 10,, 9 1, 3, 8 ml, 10 ml, 3, PL A, 10 ml 012 ml, PL A/, 10 ml 012 ml cm 2, mm 25 mm, 4 h, 10 ml, 37,,, 70 % 30 min, 012 ml,, 60 min, 5 min (750g), : 0, 1,, 545nm ( ) 2, (, ) A ( 3, ( ) 4 ) = ( A 0, 1, ( A ) / ( A A ) ) 2, ( 100 % < 5 %, 1 mm) 3, ( ; > 5 % > 1 mm, ) , , 210, > 2 cm, 419, 510, 10 %, h 37 5 min, 012 ml (ACD ), 5 min, ( kg,, 7 d ml CaCl2 (012 mol L - 1 ), ), 2 h, 1 min, CaCl2,,, 5 min, 100 ml ( 6 cm), 60 min, 10 min,, 1, 2 3, 2 < 012, 540 nm A, 2 A, min, PL A/ min A, PL A/, A 01100, PL A 3 PL A, 10 ml kg - 1,,, 37 1 h 1, 3, , < 012, PLA/ PL A, < ml SPSS 1010,,,,, 2, ,,, 211 PL A PLA/

4 408 ( ) PL A 150 m, > 90 %, ; PLA/ PL A,,,,, 1 1 PLA PLA/ ( 150) Fig1 1 Stereoscan photograph of PLA scaffolds ( A ) PLA/ silk fibroin compound scaffolds (B) ( 150) and, 1 PLA = 15150, T1 = ; PL A/ = 5150, T1 = U = , W = , Z = , P < ,, P = 2 < ,, PL A/ PL A, PL A A = 01500, A = min A,, 2, PLA/ 37 min, PL A 26 min, PL A/ PL A 212 < 5 %, 1 PLA PLA/ Tab11 Effects of PLA and PLA/ silk fibroin compound scaffolds on hemolysis rate in rabbits Group Hemolysis rate ( / %) No PL A scaffolds Compound scaffolds , PL A/, PL A, PL A ml - 1, PL A/ ml - 1, 3 2 PLA PLA/ Fig1 2 Effects of PLA and PLA/ sild fibroin compound scaffolds on dynamic coagulation time in whole blood of rabbit s 214 8, 3 PLA PLA/ Fig13 Effects of PLA (A) and PLA/ silk scoffolds fibroin compound scaffolds (B) on rabbit dermal stem cells

5 , 1 / ,, 4, PL A/ PL A, PL A/ PL A PL A PL A/, 2 2 PLA PLA/ Tab12 Effects of scoffolds on the temperature in rabbits PLA and PLA/ sick fibroin compiund ( / ) Group T (Before injection) T(after injection) ( t/ h) PLA Compound scoffolds PLA PLA/ 3 Fig1 4 Effects of PLA and PLA/ silk fibroin compound, scoffolds on the curve of dermal stem cells in rabbits, 215, 24h 6 ; 48 h 10 ; 72 h 4, 10 PL A/, 24 h 1 1, 1 1, ; 48 h, ;, PL A,,, PL A P GA FDA,,, 72 h, Altman [10 ], : PL A 24 h 2 2, 1 1, ; 48 h, ; 72 h, 72 h, 216 6, < 012, < 110,,, ;, ; ; ; Inoue [11 ] 3,, 30 % 50 %,,, [10 ]

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