Experimental study on ectopic osteogenesis by implants of gelatin sponge combined with recombinant human bone morphogenetic protein-7

Abstract

Abstract: AIM: To investigate the ectopic osteogenetic capacity of gelatin sponge contained with recombinant human bone morphogenetic protein-7 (rhBMP-7) by animal experiment and evaluate the biologic activity of rhBMP-7 expressed in Escherichia coli. METHODS: KM mice were divided into two groups randomly. Gelatin sponge contained with rhBMP-7 were implanted into intramuscular spatium of mice as experimental group, while gelatin sponge were implanted as control group. The mice were sacrificed and the tissue samples were harvested at 1st, 2nd, 3rd and 4th week after implantation. Tissue response was observed by HE staining. The contents of proteins and calcium were measured and the activities of alkaline phosphatase (ALP) were detected. RESULTS: HE staining showed that some cells similar to osteocytes and calcific tissues were formed in the group of gelatin sponge contained with rhBMP-7 after implantation at 2nd, 3rd and 4th week. The contents of proteins and calcium and the activities of ALP in experimental group were significantly higher than those in the control group which implanted with gelatin sponge at 2nd, 3rd and 4th week.CONCLUSION: The gelatin sponge combined with rhBMP-7 could induce ectopic osteogenesis more effectively and it is a good biomaterial which could be used to detect the biological activity of rhBMP-7.

Key words: Gelatin sponge, Recombinant human bone morphogenetic protein-7, Ectopic osteogenesis, Mice

CLC Number:

WANG Guo-dong, LI Ping, HAN Jin-xiang, WANG Shi-li, LIU Li-na. Experimental study on ectopic osteogenesis by implants of gelatin sponge combined with recombinant human bone morphogenetic protein-7[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(12): 1349-1354.

References

[1] Urist MR.Bone: formation by autoinduction[J]. Science, 1965, 150(3698):893-899.
[2] Cheline AJ, Reddi AH, Martin RB.Bone morphogenetic protein-7 selectively enhances mechanically induced bone formation[J]. Bone, 2002, 31(5): 570-574.
[3] Lissenberg-Thunnissen SN, de Gorter DJ, Sier CF, et al. Use and efficacy of bone morphogenetic proteins in fracture healing[J]. Int Orthop, 2011, 35(9): 1271-1280.
[4] 车婧, 韩金祥, 王世立, 等. 重组人成骨蛋白-1 在大肠杆菌中的高效表达及复性[J]. 中国药学杂志, 2005, 40(13): 1023-1026.
[5] 王国栋, 王世立, 韩金祥, 等. 表达重组人骨形态发生蛋白-7工程菌的发酵及表达产物的纯化[J]. 生物技术通讯, 2006, 17(3): 345-348.
[6] 王军志. 生物技术药物研究开发和质量控制[M]. 2版. 北京: 科学出版社, 2007: 797-798.
[7] 谈伟强, 徐靖宏, 刘友山, 等. 胶原蛋白/BMP复合材料的制备和成骨性能研究[J]. 生物工程学报, 2008, 24(2): 272-277.
[8] 李想, 董纪元, 彭江, 等. 两种材料复合 rhBMP-2诱导大鼠皮下异位成骨的比较研究[J]. 军进修学院学报, 2011, 32(3): 280-283.
[9] Ji Y, Xu GP, Zhang ZP, et al.BMP-2/PLGA delayed-release microspheres composite graft, selection of bone particulate diameters, and prevention of aseptic inflammation for bone tissue engineering[J]. Ann Biomed Eng, 2010, 38(3): 632-639.
[10] 陈顺广, 郑启新, 郭晓东, 等. TGF -β1 基因转染骨髓基质干细胞促进 BMP-2活性多肽大鼠体内异位成骨[J].华中科技大学学报:医学版, 2009, 38(1): 44-48.
[11] Boden SD.The ABCs of BMPs[J]. Orthop Nurs,2005,24(1):49-54.
[12] Asahina I, Sampath TK, Hauschka PV.Human osteogenic protein-1 induces chondroblastic, osteoblastic, and/or adipocytic differentiation of clonal murine target cells[J]. Exp Cell Res, 1996, 222(1): 38-47.
[13] Schofer MD, Veltum A, Theisen C, et al.Functionalisation of PLLA nanofiber scaffolds using a possible cooperative effect between collagen type I and BMP-2: impact on growth and osteogenic differentiation of human mesenchymal stem cells[J]. J Mater Sci Mater Med, 2011, 22(7): 1753-1762.
[14] Coelho MJ, Cabral AT, Fernnades MH.Human bone cell cultures in biocompatibility testing. Part I:osteoblastic dfierentiation of seirally passaged human bone mrrow cells cultured in α-MEM and in DMEM[J]. Biomaterials,2000,21(11):1087-1094.
[15] 蒋月桂, 孙斌, 苟建重, 等. rhBMP-2HACo复合材料异位诱导成骨中碱性磷酸酶活性的检测[J]. 中国美容医学, 2010, 19(4): 521-523.
[16] 李高舜, 叶招明, 杨迪生. 木脂素分解产物 enterolactone 和 enterodiol对成骨细胞样细胞 MG63 的双相作用[J]. 中国临床药理学与治疗学, 2011, 16(1): 46-50.