FABRICATION AND BIOCOMPATIBILITY EVALUATION OF POLYURETHANE-ACELLULAR MATRIX COMPOSITE SCAFFOLD IN VITRO AND IN VIVO_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

XIAOYuan ^1,2 , ZHANGJie ² , LUYanrong ² , YUANHaichao ³ , BAILin ² ,  JIANGXia ² ,  CHENGJingqiu ²

1. College of Life Sciences, Sichuan University, Chengdu Sichuan, 610065, P. R. China;
2. Regenerative Medicine Research Center;
3. Department of Urology, West China Hospital, Sichuan University, Chengdu Sichuan, 610065, P. R. China;

Corresponding?author：

JIANGXia, Email: jiangxia52@163.com; CHENGJingqiu, Email: jqcheng@scu.edu.cn

Keywords：

Acellular matrix; Polyurethane; Bladder repair; Tissue engineering; Rabbit

DOI：

10.7507/1002-1892.20150217

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Objective To prepare a composite scaffold using bladder acellular matrix (BACM) and polyurethane (PU) for bladder repair and regeneration, and to evaluate its mechanical properties and biocompatibility. Methods Fresh bladder tissues were obtained from New Zealand rabbits and then treated with 1%SDS and 1%Triton X-100 to obtain BACM. The BACM was combined with PU to fabricate PU-BACM composite scaffold. The tensile strength and elongation at break of BACM and PU-BACM scaffolds were tested. Scaffolds and extracts of scaffolds were prepared to evaluate the biocompatibility. For cell-proliferation analysis, cell counting kit 8 method was used at 1, 3, 5, and 7 days after co-culture of human bladder smooth muscle cell (HBSMC) and scaffolds. The cell cycle was tested by flow cytometry after HBSMC co-cultured with extracts of scaffolds and DMEM culture medium (control group) for 24 hours. Finally, 12 New Zealand rabbits were used to establish the model of bladder repair and regeneration. Incision of 5 mm was made on the bladder, and PU-BACM scaffold was sutured with the incision. The rabbits were sacrificed at 10, 20, 40, and 60 days after surgery to observe the inflammatory cell infiltration, new tissues formation, and regeneration of epithelium by HE staining. Results The tensile strength of BACM and PU-BACM composite scaffold was (5.78 ± 0.85) N and (11.88 ± 3.21) N, and elongation at break was 14.46%±3.21% and 23.14%±1.32% respectively, all showing significant diffeence (t=3.182, P=0.034；t=4.332, P=0.012). The cell-proliferation rates of controls, PU, BACM, and PU-BACM were 36.78%±1.21%, 30.49%±0.89%, 18.92%±0.84%, and 22.42%±1.55%, it was significantly higher in PU-BACM than BACM (P<0.05). In the bladder repair and regeneration experiment, inflammatory cell infiltration was observed at 10 days after operation, and reduced at 20 days after implantation. In the meanwhile, the degradation of scaffolds was observed in vivo. The regeneration of epithelium could be observed after 40 days of implantation. At 60 days after implantation, in situ bladder tissue formed. Conclusion PU-BACM composite scaffold has higher mechanical properties and better biocompatibility than BACM scaffold. PU-BACM composite scaffold will not lead to strong immune response, and new bladder tissue can form in the in vivo rabbit bladder repair experiment. These results can provide research basis and theoretical data for further study.

Citation： XIAOYuan, ZHANGJie, LUYanrong, YUANHaichao, BAILin, JIANGXia, CHENGJingqiu. FABRICATION AND BIOCOMPATIBILITY EVALUATION OF POLYURETHANE-ACELLULAR MATRIX COMPOSITE SCAFFOLD IN VITRO AND IN VIVO. Chinese Journal of Reparative and Reconstructive Surgery, 2015, 29(8): 1016-1021. doi: 10.7507/1002-1892.20150217 Copy

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13.	劉國鋒,何志娟,楊大平,等.利用纖維蛋白膠復合血管脫細胞基質制備全生物型小口徑組織工程血管支架材料的研究.中國康復理論與實踐,2010,16(8):748-751.
14.	蔣曉瓊,熊先澤,程南生,等.去細胞支架材料用于膽管替代的相容性研究.中國普外基礎與臨床雜志,2010,17(3):248-252.
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16.	Tu DD,Chung YG,Gil ES,et al.Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty.Biomaterials,2013,34(34):8681-8689.
17.	Han DK,Park K,Park KD,et al.In vivo biocompatibility of sulfonated pEO-grafted polyurethanes for polymer heart valve and vascular graft.Artif Organs,2006,30(12):955-959.
18.	王振顯,龐國勛,宋永周,等.人脫細胞羊膜移植物用于兔膀胱切除修復的研究.實用醫學雜志,2009,25(12):1913-1915.

1. Stanasel I,Mirirzazadeh M,Smith JJ 3rd.Bladder tissue engineering.Urol Clin North Am,2010,37(4):593-599.
2. Atala A.Tissue engineering of human bladder.Br Med Bull,2011,97:81-104.
3. 祁娜,黎麗茜,田虹.組織工程膀胱支架材料:應用現狀及血管化策略.中國組織工程研究,2014,18(47):7659-7665.
4. Engelhardt EM,Micol LA,Houis S,et al.A collagen-poly (lactic acid-co-ε-caprolactone) hybrid scaffold for bladder tissue regeneration.Biomaterials,2011,32(16):3969-3796.
5. Ahvaz HH,Soleimani M,Mobasheri H,et al.Effective combination of hydrostatic pressure and aligned nanofibrous scaffolds on human bladder smooth muscle cells:implication for bladder tissue engineering.J Mater Sci Mater Med,2012,23(9):2281-2290.
6. 員海超,唐寅,白云金,等.自體骨髓間充質干細胞復合膀胱去細胞基質修復犬膀胱缺損的實驗研究.四川大學學報:醫學版,2015,46(1):1-5.
7. 李珍美玉,顧蕓,異晟.細胞外基質在組織工程中的應用.交通醫學,2014,28(5):425-432.
8. Boruch AV,Nieponice A,Qureshi IR,et al.Constructive remodeling of biologic scaffolds is dependent on early exposure to physiologic bladder filling in a canine partial cystectomy model.J Surg Res,2012,161(2):217-225.
9. Pereira IM,Axisa F,Oréfice RL,et al.Shape-memory anchoring system for bladder sensors.J Biomed Mater Res B Appl Biomater,2011,96(2):369-375.
10. Jiang X,Yu F,Wang Z,et al.Fabrication an characterization of waterborne biodegradable polyurethanes 3-dimensional porous scaffolds for vascular tissue engineering.J Biomater Sci Polym Ed,2012,21(12):1637-1652.
11. Jiang X,Wang K,Ding M,et al.Quantitative grafting of peptide onto the nontoxic biodegradable waterborne polyurethanes to fabricate peptide modified scaffold for soft tissue engineering.J Mater Sci Mater Med,2011,22(4):819-827.
12. Jiang X,Li JH,Ding MM,et al.Synthesis and degradation of nontoxic biodegradable waterborne polyurethanes elastomer with poly (epsilon-caprolactone) and poly (ethylene glycol) as soft segment.Eur Polym J,2007,43(5):1838-1846.
13. 劉國鋒,何志娟,楊大平,等.利用纖維蛋白膠復合血管脫細胞基質制備全生物型小口徑組織工程血管支架材料的研究.中國康復理論與實踐,2010,16(8):748-751.
14. 蔣曉瓊,熊先澤,程南生,等.去細胞支架材料用于膽管替代的相容性研究.中國普外基礎與臨床雜志,2010,17(3):248-252.
15. Jang YJ,Chun SY,Kim GN,et al.Characterization of a novel composite scaffold consisting of acellular bladder submucosa matrix,polycaprolactone and Pluronic F127 as a substance for bladder reconstruction.Acta Biomater,2014,10(4):3117-3125.
16. Tu DD,Chung YG,Gil ES,et al.Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty.Biomaterials,2013,34(34):8681-8689.
17. Han DK,Park K,Park KD,et al.In vivo biocompatibility of sulfonated pEO-grafted polyurethanes for polymer heart valve and vascular graft.Artif Organs,2006,30(12):955-959.
18. 王振顯,龐國勛,宋永周,等.人脫細胞羊膜移植物用于兔膀胱切除修復的研究.實用醫學雜志,2009,25(12):1913-1915.

Chinese Journal of Reparative and Reconstructive Surgery

FABRICATION AND BIOCOMPATIBILITY EVALUATION OF POLYURETHANE-ACELLULAR MATRIX COMPOSITE SCAFFOLD IN VITRO AND IN VIVO

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