Experimental study on long-term outcome of porcine collagen membrane xenotransplantation <i>in vivo</i> _Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Weidong ,  XIE Weiguo , ZHAO Chaoli , WANG Hui , YE Ziqing

Department of Burns, the Third Hospital of Wuhan, Tongren Hospital of Wuhan University, Wuhan Hubei, 430060, P.R.China;

Corresponding author：

XIE Weiguo, Email: wgxie@hotmail.com

Keywords：

Porcine collagen membrane; biocompatibility; biological material; xenotransplantion; rat

DOI：

10.7507/1002-1892.201708123

Video：

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ObjectiveTo observe the long-term outcome and biocompatibility of the porcine collagen membrane (DermalGen) after xenotransplantation in vivo.MethodsTwenty Sprague Dawley rats were randomly divided into 2 groups (n=10). DermalGen were implanted subcutaneously into the dorsum of rats in experimental group, and the rats in control group were treated with sham-operation. At 3, 7, and 15 days and 1, 3, 6, and 12 months after operation, the samples of experimental group were harvested and gross observation, histological observation, CD31 immunohistochemical staining, and transmission electron microscope observation were taken to observe the inflammatory reaction, angiogenesis, and collagen arrangement. The skin tissues of the control group at 12 months were observed and compared.ResultsAll incisions healed in experimental group, without obvious swelling and inflammatory reaction. The DermalGen was closely contact with the surrounding tissue without obvious rejection, and it was still legible at 12 months. Histological observation of experimental group showed that the infiltration of fibroblasts and inflammatory cells were seen at 7 days. More capillaries and fibroblast cells were seen and the inflammatory response gradually faded at 15 days and 1 month. There were abundant vessels and cells in the DermalGen at 3 months. The angiogenesis and fibroblasts decreased gradually, and the collagen started to format and margin blended simultaneously at 6 and 12 months. The inflammatory cells in experimental group at 15 days and 1 month were significantly more than that in control group (P<0.05), and no significant difference was found at 12 months between experimental group and control group (P>0.05). Immunohistochemical staining of experimental group showed that the angiogenesis changed obviously with the time, and the density of vessels decreased significantly at 12 months. Compared with control group, the possitive expressions of CD31 in experimental group at 15 days and 12 months after operation were signiﬁcantly decreased (P<0.05), and were significantly increased at 1 month (P<0.05). Transmision electron microscope observation showed that the arrangement of collagen in grafted DermalGen had no obvious changed when compared with the DermalGen, and vascular endothelial cell, capillarypericytes and fibroblast cells could be seen inside.ConclusionThe DermalGen structure is stable after long-term xenotransplantation and with good tolerogenic property in vivo.

Citation： ZHANG Weidong, XIE Weiguo, ZHAO Chaoli, WANG Hui, YE Ziqing. Experimental study on long-term outcome of porcine collagen membrane xenotransplantation in vivo . Chinese Journal of Reparative and Reconstructive Surgery, 2018, 32(4): 462-467. doi: 10.7507/1002-1892.201708123 Copy

1.	Livesey SA, Herndon DN, Hollyyoak MA, et al. Transplanted acellular allograft dermal matrix. Potential as a template for the reconstruction of viable dermis. Transplantation, 1995, 60(1): 1-9.
2.	唐修俊, 李丰, 魏在荣, 等. 异体脱细胞真皮基质联合自体刃厚皮移植修复供瓣区 20 例. 中华烧伤杂志, 2015, 31(3): 226-228.
3.	陆莹莹, 许争, 温东朋, 等. 脱细胞真皮基质修复组织缺损研究进展. 国际生物医学工程杂志, 2017, 40(1): 58-61.
4.	赵鹏, 郝利明, 贾志刚, 等. 生物护创膜的理化性质及对大鼠深 Ⅱ 度烫伤创面愈合的影响. 中华烧伤杂志, 2015, 31(1): 52-55.
5.	周敏, 王信春, 石兵, 等. 无菌生物护创膜在烧伤创面的应用. 四川医学, 2014, 35(8): 1077-1078.
6.	过云, 蔡良良, 虞俊杰, 等. 无菌生物护创膜治疗小儿深 Ⅱ 度烧伤创面的临床观察. 中华损伤与修复杂志 (电子版), 2013, 8(1): 56-57.
7.	綦惠, 杰永生, 陈磊, 等. 脱细胞真皮基质制备及其生物相容性研究. 中国修复重建外科杂志, 2014, 28(6): 768-772.
8.	廖婷, 邹瑞, 闫璟, 等. 脱细胞真皮基质移植后组织学转归的实验研究. 实用医学杂志, 2016, 32(11): 1749-1752.
9.	张卫东, 谢卫国, 赵超莉, 等. 护创膜在兔微粒皮移植术中的作用研究. 中华烧伤杂志, 2012, 28(5): 336-340.
10.	Franz S, Allenstein F, Kajahn J, et al. Artificial extracellular matrices composed of collagenⅠand high-sulfated hyaluronan promote phenotypic and functional modulation of human pro-inflammatory M1 macrophages. Acta Biomater, 2013, 9(3): 5621-5629.
11.	Silva JJ, Pompeu DG, Ximenes NC, et al. Effects of kaurenoic acid and arginine on random skin flap oxidative stress. Inflammation, and cytokines in rats. Aesthetic Plast Surg, 2015, 39(6): 971-977.
12.	谢卫国, 谭红, 赵超莉, 等. 真皮替代物移植后的血管化过程及组织学变化的实验研究. 中华烧伤杂志, 2005, 21(1): 37-39.
13.	Pascual G, Sotomayor S, Adel F, et al. Remodeling of noncrosslinked acellular dermal datrices in a rabbit model of ventral hernia repair. Eur Surg Res, 2015, 56(1-2): 32-48.
14.	Fioretti F, Lebreton-DeCoster C, Gueniche F, et al. Human bone marrow-derived cells: an attractive source to populate dermal substitutes. Wound Repair Regen, 2008, 16(1): 87-94.
15.	Anjum F, Agabalyan NA, Sparks HD, et al. Biocomposite nanofiber matrices to support ECM remodeling by human dermal progenitors and enhanced wound closure. Sci Rep, 2017, 7(1): 10291.
16.	Wang N, Liu H, Li X, et al. Activities of MSCs derived from transgenic mice seeded on ADM scaffolds in wound healing and assessment by advanced optical techniques. Cell Physiol Biochem, 2017, 42(2): 623-639.
17.	林才, 罗旭, 王平, 等. 激光微孔猪脱细胞真皮基质的制备及生物相容性评价.中华烧伤杂志, 2011, 27(6): 463-465.
18.	Bonvallet PP, Schultz MJ, Mitchell EH, et al. Microporous dermal-mimetic electrospun scaffolds pre-seeded with fibroblasts promote tissue regeneration in full-thickness skin wounds. PLoS One, 2015, 10(3): e0122359.
19.	张迪, 谭秋雯, 汤沈力, 等. 一种优化的猪骨骼肌来源无细胞基质支架材料制备方法及鉴定. 中国修复重建外科杂志, 2016, 30(10): 1282-1289.

1. Livesey SA, Herndon DN, Hollyyoak MA, et al. Transplanted acellular allograft dermal matrix. Potential as a template for the reconstruction of viable dermis. Transplantation, 1995, 60(1): 1-9.
2. 唐修俊, 李丰, 魏在荣, 等. 异体脱细胞真皮基质联合自体刃厚皮移植修复供瓣区 20 例. 中华烧伤杂志, 2015, 31(3): 226-228.
3. 陆莹莹, 许争, 温东朋, 等. 脱细胞真皮基质修复组织缺损研究进展. 国际生物医学工程杂志, 2017, 40(1): 58-61.
4. 赵鹏, 郝利明, 贾志刚, 等. 生物护创膜的理化性质及对大鼠深 Ⅱ 度烫伤创面愈合的影响. 中华烧伤杂志, 2015, 31(1): 52-55.
5. 周敏, 王信春, 石兵, 等. 无菌生物护创膜在烧伤创面的应用. 四川医学, 2014, 35(8): 1077-1078.
6. 过云, 蔡良良, 虞俊杰, 等. 无菌生物护创膜治疗小儿深 Ⅱ 度烧伤创面的临床观察. 中华损伤与修复杂志 (电子版), 2013, 8(1): 56-57.
7. 綦惠, 杰永生, 陈磊, 等. 脱细胞真皮基质制备及其生物相容性研究. 中国修复重建外科杂志, 2014, 28(6): 768-772.
8. 廖婷, 邹瑞, 闫璟, 等. 脱细胞真皮基质移植后组织学转归的实验研究. 实用医学杂志, 2016, 32(11): 1749-1752.
9. 张卫东, 谢卫国, 赵超莉, 等. 护创膜在兔微粒皮移植术中的作用研究. 中华烧伤杂志, 2012, 28(5): 336-340.
10. Franz S, Allenstein F, Kajahn J, et al. Artificial extracellular matrices composed of collagenⅠand high-sulfated hyaluronan promote phenotypic and functional modulation of human pro-inflammatory M1 macrophages. Acta Biomater, 2013, 9(3): 5621-5629.
11. Silva JJ, Pompeu DG, Ximenes NC, et al. Effects of kaurenoic acid and arginine on random skin flap oxidative stress. Inflammation, and cytokines in rats. Aesthetic Plast Surg, 2015, 39(6): 971-977.
12. 谢卫国, 谭红, 赵超莉, 等. 真皮替代物移植后的血管化过程及组织学变化的实验研究. 中华烧伤杂志, 2005, 21(1): 37-39.
13. Pascual G, Sotomayor S, Adel F, et al. Remodeling of noncrosslinked acellular dermal datrices in a rabbit model of ventral hernia repair. Eur Surg Res, 2015, 56(1-2): 32-48.
14. Fioretti F, Lebreton-DeCoster C, Gueniche F, et al. Human bone marrow-derived cells: an attractive source to populate dermal substitutes. Wound Repair Regen, 2008, 16(1): 87-94.
15. Anjum F, Agabalyan NA, Sparks HD, et al. Biocomposite nanofiber matrices to support ECM remodeling by human dermal progenitors and enhanced wound closure. Sci Rep, 2017, 7(1): 10291.
16. Wang N, Liu H, Li X, et al. Activities of MSCs derived from transgenic mice seeded on ADM scaffolds in wound healing and assessment by advanced optical techniques. Cell Physiol Biochem, 2017, 42(2): 623-639.
17. 林才, 罗旭, 王平, 等. 激光微孔猪脱细胞真皮基质的制备及生物相容性评价.中华烧伤杂志, 2011, 27(6): 463-465.
18. Bonvallet PP, Schultz MJ, Mitchell EH, et al. Microporous dermal-mimetic electrospun scaffolds pre-seeded with fibroblasts promote tissue regeneration in full-thickness skin wounds. PLoS One, 2015, 10(3): e0122359.
19. 张迪, 谭秋雯, 汤沈力, 等. 一种优化的猪骨骼肌来源无细胞基质支架材料制备方法及鉴定. 中国修复重建外科杂志, 2016, 30(10): 1282-1289.

Chinese Journal of Reparative and Reconstructive Surgery

Experimental study on long-term outcome of porcine collagen membrane xenotransplantation in vivo

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