BIOCOMPATIBILITY OF POROUS POLY LACTIC ACID/BONE MATRIX GELATIN COMPOSITE BIOMATERIALS FOR BONE REPAIR_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANGYumin ¹ ,  LIJing ² , NIUXiaojun ¹ , LIUJianchun ¹ , WANGJue ¹ , GAOLan ¹

1. Basic Medical School, Shanxi University of Traditional Chinese Medicine, Taiyuan Shanxi, 030024, P. R. China;
2. China Institute for Radiation Protection;

Corresponding author：

LIJing, Email: 9985855@qq.com

Keywords：

Poly lactic acid; Bone matrix gelatin; Biocompatibility; Bone substitute; Rat

DOI：

10.7507/1002-1892.20160050

Video：

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Objective To evaluate the biocompatibility of poly lactic acid/bone matrix gelatin (PLA/BMG) composite biomaterial so as to lay a foundation for bone defect repair. Methods Rats'MC3T3-E1 cells were cultured with leaching solution of PLA/BMG and PLA material respectively for 7 days. The cell proliferation rate was tested by MTT and cell toxicity grading was carried out everyday. The PLA/BMG and MC3T3-E1 cells were co-cultured, the cell shape and proliferation were observed by inverted phase contrast microscope at 1, 3, and 5 days and cell adhesion by scanning electron microscope at 5 days. The PLA and PLA/BMG were implanted subcutaneously in 15 Wistar rats. The histological observation was done, and the thickness of fibrous membrane, the number of inflammatory cells, and the vascularization area were measured at postoperative 2nd, 4th, and 8th week. Results The tests for cytotoxicity in vitro showed that the cell proliferation rates were over 100% and the cell cytotoxic grades were grade 0 at 1-7 days in PLA/BMG group. While in PLA group, the cell proliferation rates were less than 100% and the cell cytotoxic grades were grade 1 at 2, 4, and 7 days. After co-culture of PLA/BMG and MC3T3-E1 cells, cells grew on the surface and in the pores of PLA/BMG, and the cellular morphology was triangle or polygon with abundant microvillus on the surface. After subcutaneous implantation, the rats survived to the end of experiment, and incision healed well. PLA was wrapped by connective tissue where there were a lot of lymphocytes and neutrophilic granulocytes. The cells and tissue grew slowly in PLA. The PLA/BMG materials were wrapped by little connective tissue where there were a few inflammatory cells. The connective tissue ingrowth was observed in the center of PLA/BMG. There was no significant difference in the thickness of fibrous membrane between 2 groups at each time point (P>0.05). The number of inflammatory cells of PLA/BMG group were significantly less than those in PLA group at 2, 4, and 8 weeks (P<0.05); the vascularization area was significantly larger than that in PLA group (P<0.05). Conclusion PLA/BMG composite biomaterials prepared by super critical-CO₂ technique are good in cell and tissue biocompatibilty.

Citation： ZHANGYumin, LIJing, NIUXiaojun, LIUJianchun, WANGJue, GAOLan. BIOCOMPATIBILITY OF POROUS POLY LACTIC ACID/BONE MATRIX GELATIN COMPOSITE BIOMATERIALS FOR BONE REPAIR. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(2): 251-257. doi: 10.7507/1002-1892.20160050 Copy

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3.	Félix Lanao RP, Jonker AM, Wolke JG, et al. Physicochemical properties and applications of poly (lactic-co-glycolic acid) for use in bone regeneration. Tissue Eng Part B Rev, 2013, 19(4):380-390.
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12.	黄金龙, 夏虹, 李丽华, 等. 异种骨与成骨细胞体外生物相容性研究. 中国矫形外科杂志, 2014, 22(2):158-162.
13.	Trang NT, Chinhl NT, Thanh DT, et al. Investigating the properties and hydrolysis ability of poly-lactic acid/chitosan nanocomposites using polycaprolactone. J Nanosci Nanotechnol, 2015, 15(12):9585-9590.
14.	Manavitehrani I, Fathi A, Wang Y, et al. Reinforced poly (propylene carbonate) composite with enhanced and tunable characteristics, an alternative for poly (lactic acid). ACS Appl Mater Interfaces, 2015, 7(40):22421-22430.
15.	Russias J, Saiz E, Nalla RK, et al. Fabrication and mechanical properties of PLA/HA composites:A study of in vitro degradation. Mater Sci Eng C Biomim Supramol Syst, 2006, 26(8):1289-1295.
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17.	Ullm S, Krüger A, Tondera C, et al. Biocompatibility and inflammatory response in vitro and in vivo to gelatin-based biomaterials with tailorable elastic properties. Biomaterials, 2014, 35(37):9755-9766.
18.	Valiense H, Barreto M, Resende RF, et al. In vitro and in vivo evaluation of strontium-containing nanostructured carbonated hydroxyapatite/sodium alginate for sinus lift in rabbits. J Biomed Mater Res B Appl Biomater, 2016, 104(2):274-282.
19.	连芩, 庄佩, 李常海, 等. 3-D打印双管道聚乳酸/β-磷酸三钙生物陶瓷复合材料支架的力学性能研究. 中国修复重建外科杂志, 2014, 28(3):309-313.
20.	Lopes MS, Jardini AL. Poly (lactic acid) production for tissue engineering application. Procedia Engineering, 2012, 42:1402-1413.
21.	Włodarski P, Galus R, Brodzikowska A, et al. Osteogenic efficiency of demineralized and lyophylized xenogeneic bone and syngeneic dentine implants in mice. Folia Biol (Krakow), 2013, 61(1-2):25-29.
22.	Bigham AS, Shadkhast M, Bigham Sadegh A, et al. Evaluation of osteoinduction properties of the demineralized bovine foetal growth plate powder as a new xenogenic biomaterial in rat. Res Vet Sci, 2011, 91(2):306-310.

1. Miyazaki T, Ishikawa K, Shirosaki Y, et al. Organic-inorganic composites designed for biomedical applications. Biol Pharm Bull, 2013, 36(11):1670-1675.
2. Jung UW, Lee JS, Lee G, et al. Role of collagen membrane in lateral onlay grafting with bovine hydroxyapatite incorporated with collagen matrix in dogs. J Periodontal Implant Sci, 2013, 43(2):64-71.
3. Félix Lanao RP, Jonker AM, Wolke JG, et al. Physicochemical properties and applications of poly (lactic-co-glycolic acid) for use in bone regeneration. Tissue Eng Part B Rev, 2013, 19(4):380-390.
4. Delabarde C, Plummer CJ, Bourban PE, et al. Biodegradable polylactide/hydroxyapatite nanocomposite foam scaffolds for bone tissue engineering applications. J Mater Sci Mater Med, 2012, 23(6):1371-1385.
5. 张育敏, 李宝兴, 李冀, 等. 聚乳酸-骨基质明胶多孔复合材料制备及骨诱导活性研究. 中国修复重建外科杂志, 2007, 21(2):135-139.
6. 中华人民共和国国家标准医疗器械生物学评价第5部分:细胞毒性试验:体外法. 北京:中国标准出版社, 1997.
7. 卫生部药政管理局. 生物材料和医疗器材生物学评价技术要求. 北京:中国标准出版社, 1997:1-44.
8. 郝和平, 卜长生, 刘秦玉, 等. 医疗器械生物学评价标准实施指南. 北京:中国标准出版社, 2002.
9. Bruinink A, Luginbuehl R. Evaluation of biocompatibility using in vitro methods:interpretation and limitations. Adv Biochem Eng Biotechnol, 2012, 126:117-152.
10. Mangala MG, Chandra SM, Bhavle RM. To evaluate the biocompatibility of the Indian Portland cement with potential for use in dentistry:An animal study. J Conserv Dent, 2015, 18(6):440-444.
11. Di Virgilio AL, Arnal PM, Maisuls I. Biocompatibility of core@shell particles:cytotoxicity and genotoxicity in human osteosarcoma cells of colloidal silica spheres coated with crystalline or amorphous zirconia. Mutat Res Genet Toxicol Environ Mutagen, 2014, 770:85-94.
12. 黄金龙, 夏虹, 李丽华, 等. 异种骨与成骨细胞体外生物相容性研究. 中国矫形外科杂志, 2014, 22(2):158-162.
13. Trang NT, Chinhl NT, Thanh DT, et al. Investigating the properties and hydrolysis ability of poly-lactic acid/chitosan nanocomposites using polycaprolactone. J Nanosci Nanotechnol, 2015, 15(12):9585-9590.
14. Manavitehrani I, Fathi A, Wang Y, et al. Reinforced poly (propylene carbonate) composite with enhanced and tunable characteristics, an alternative for poly (lactic acid). ACS Appl Mater Interfaces, 2015, 7(40):22421-22430.
15. Russias J, Saiz E, Nalla RK, et al. Fabrication and mechanical properties of PLA/HA composites:A study of in vitro degradation. Mater Sci Eng C Biomim Supramol Syst, 2006, 26(8):1289-1295.
16. Persson M, Lorite GS, Kokkonen HE, et al. Effect of bioactive extruded PLA/HA composite films on focal adhesion formation of preosteoblastic cells. Colloids Surf B Biointerfaces, 2014, 1(121):409-416.
17. Ullm S, Krüger A, Tondera C, et al. Biocompatibility and inflammatory response in vitro and in vivo to gelatin-based biomaterials with tailorable elastic properties. Biomaterials, 2014, 35(37):9755-9766.
18. Valiense H, Barreto M, Resende RF, et al. In vitro and in vivo evaluation of strontium-containing nanostructured carbonated hydroxyapatite/sodium alginate for sinus lift in rabbits. J Biomed Mater Res B Appl Biomater, 2016, 104(2):274-282.
19. 连芩, 庄佩, 李常海, 等. 3-D打印双管道聚乳酸/β-磷酸三钙生物陶瓷复合材料支架的力学性能研究. 中国修复重建外科杂志, 2014, 28(3):309-313.
20. Lopes MS, Jardini AL. Poly (lactic acid) production for tissue engineering application. Procedia Engineering, 2012, 42:1402-1413.
21. Włodarski P, Galus R, Brodzikowska A, et al. Osteogenic efficiency of demineralized and lyophylized xenogeneic bone and syngeneic dentine implants in mice. Folia Biol (Krakow), 2013, 61(1-2):25-29.
22. Bigham AS, Shadkhast M, Bigham Sadegh A, et al. Evaluation of osteoinduction properties of the demineralized bovine foetal growth plate powder as a new xenogenic biomaterial in rat. Res Vet Sci, 2011, 91(2):306-310.

Chinese Journal of Reparative and Reconstructive Surgery

BIOCOMPATIBILITY OF POROUS POLY LACTIC ACID/BONE MATRIX GELATIN COMPOSITE BIOMATERIALS FOR BONE REPAIR

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