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 signi?cantly 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, 李豐, 魏在榮, 等. 異體脫細胞真皮基質聯合自體刃厚皮移植修復供瓣區 20 例. 中華燒傷雜志, 2015, 31(3): 226-228.
3.	陸瑩瑩, 許爭, 溫東朋, 等. 脫細胞真皮基質修復組織缺損研究進展. 國際生物醫學工程雜志, 2017, 40(1): 58-61.
4.	趙鵬, 郝利明, 賈志剛, 等. 生物護創膜的理化性質及對大鼠深 Ⅱ 度燙傷創面愈合的影響. 中華燒傷雜志, 2015, 31(1): 52-55.
5.	周敏, 王信春, 石兵, 等. 無菌生物護創膜在燒傷創面的應用. 四川醫學, 2014, 35(8): 1077-1078.
6.	過云, 蔡良良, 虞俊杰, 等. 無菌生物護創膜治療小兒深 Ⅱ 度燒傷創面的臨床觀察. 中華損傷與修復雜志 (電子版), 2013, 8(1): 56-57.
7.	綦惠, 杰永生, 陳磊, 等. 脫細胞真皮基質制備及其生物相容性研究. 中國修復重建外科雜志, 2014, 28(6): 768-772.
8.	廖婷, 鄒瑞, 閆璟, 等. 脫細胞真皮基質移植后組織學轉歸的實驗研究. 實用醫學雜志, 2016, 32(11): 1749-1752.
9.	張衛東, 謝衛國, 趙超莉, 等. 護創膜在兔微粒皮移植術中的作用研究. 中華燒傷雜志, 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.	謝衛國, 譚紅, 趙超莉, 等. 真皮替代物移植后的血管化過程及組織學變化的實驗研究. 中華燒傷雜志, 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, 羅旭, 王平, 等. 激光微孔豬脫細胞真皮基質的制備及生物相容性評價.中華燒傷雜志, 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.	張迪, 譚秋雯, 湯沈力, 等. 一種優化的豬骨骼肌來源無細胞基質支架材料制備方法及鑒定. 中國修復重建外科雜志, 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, 李豐, 魏在榮, 等. 異體脫細胞真皮基質聯合自體刃厚皮移植修復供瓣區 20 例. 中華燒傷雜志, 2015, 31(3): 226-228.
3. 陸瑩瑩, 許爭, 溫東朋, 等. 脫細胞真皮基質修復組織缺損研究進展. 國際生物醫學工程雜志, 2017, 40(1): 58-61.
4. 趙鵬, 郝利明, 賈志剛, 等. 生物護創膜的理化性質及對大鼠深 Ⅱ 度燙傷創面愈合的影響. 中華燒傷雜志, 2015, 31(1): 52-55.
5. 周敏, 王信春, 石兵, 等. 無菌生物護創膜在燒傷創面的應用. 四川醫學, 2014, 35(8): 1077-1078.
6. 過云, 蔡良良, 虞俊杰, 等. 無菌生物護創膜治療小兒深 Ⅱ 度燒傷創面的臨床觀察. 中華損傷與修復雜志 (電子版), 2013, 8(1): 56-57.
7. 綦惠, 杰永生, 陳磊, 等. 脫細胞真皮基質制備及其生物相容性研究. 中國修復重建外科雜志, 2014, 28(6): 768-772.
8. 廖婷, 鄒瑞, 閆璟, 等. 脫細胞真皮基質移植后組織學轉歸的實驗研究. 實用醫學雜志, 2016, 32(11): 1749-1752.
9. 張衛東, 謝衛國, 趙超莉, 等. 護創膜在兔微粒皮移植術中的作用研究. 中華燒傷雜志, 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. 謝衛國, 譚紅, 趙超莉, 等. 真皮替代物移植后的血管化過程及組織學變化的實驗研究. 中華燒傷雜志, 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, 羅旭, 王平, 等. 激光微孔豬脫細胞真皮基質的制備及生物相容性評價.中華燒傷雜志, 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. 張迪, 譚秋雯, 湯沈力, 等. 一種優化的豬骨骼肌來源無細胞基質支架材料制備方法及鑒定. 中國修復重建外科雜志, 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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