EFFECT OF ADIPOSE-DERIVED STEM CELLS COMBINED WITH CHITOSAN ON IMMEDIATE RETRACTION RATE OF EXPANDED SKIN IN RABBIT_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

YAOYongming ¹ , ZHANGZemin ¹ , YANHe ² , WUCaifeng ¹ , HUFei ¹ ,  YANGBiaobing ¹

1. Weifang Medical University, Weifang Shandong, 261053, P. R. China;
2. Department of Plastic Surgery, Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine;

Corresponding author：

YANGBiaobing, Email: ybiaobing@163.com

Keywords：

Adipose-derived stem cells; Chitosan; Expanded skin; Immediate retraction; Rabbit

DOI：

10.7507/1002-1892.20150329

Video：

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Objective To investigate the effect of adipose-derived stem cells (ADSCs) combined with chitosan on the immediate retraction rate of rabbit expanded skin. Methods ADSCs were isolated from rabbit fresh fat under sterile conditions and cultured to the 3rd generation by methods of enzymatic digestion; the specific surface markers and the differentiation into epidermal cells and cartilage cells were identified. Forty New Zealand white rabbits (aged, 2-3 months) were randomly divided into 4 groups (n=10): control group (group A), ADSCs group (group B), chitosan group (group C), and ADSCs+chitosan group (group D). ADSCs cell suspension with the concentration of 5×10⁶ cells/mL was prepared. The skin expansion model was made by embedding 30 mL dilator into the back of rabbit. Chitosan (2%, 5 mL) was coated on the surface of the dilator in groups C and D, and ADSCs cell suspension (1 mL) was injected into the skin in groups B and D. Conventional tissue expansion was performed to expected capacity at 4 weeks, and maintained for 1 week. The expanded skin was harvested to measure the immediate retraction rate, and the thickness of skin, epidermis, and fibrous capsule with HE staining. Masson staining was used to observe the characteristics of collagen in the fibrous capsule, and immunohistochemical staining for CD31 to determine the microvessel density (MVD). Results ADSCs were successfully isolated, and had multiple differentiation ability. All the animals survived to the end of the experiment. The immediate retraction rate of group D was significantly lower than that of the other groups (P<0.05), groups B and C were significantly lower than group A (P<0.05), and group B was significantly lower than group C (P<0.05). The histological staining revealed that there were more mature fibroblasts and coarse collagen fibers with regular arrangement in groups A and B; there were more naive fibroblasts and tiny and sparse collagen fibers in groups C and D. The thickness of skin and epidermis, and MVD of groups B and D were significantly larger than those of groups A and C (P<0.05); the thickness of fibrous capsule of groups C and D was significantly less than that of groups A and B (P<0.05); but no significant difference was found in the above indexes between other groups (P>0.05). Conclusion ADSCs can promote angiogenesis and regeneration of the expanded skin, have no effect on the fibrous capsule. Chitosan can inhibit the proliferation of fibrous capsule, so a combination of ADSCs and chitosan can inhibit the immediate retraction of the expanded skin.

Citation： YAOYongming, ZHANGZemin, YANHe, WUCaifeng, HUFei, YANGBiaobing. EFFECT OF ADIPOSE-DERIVED STEM CELLS COMBINED WITH CHITOSAN ON IMMEDIATE RETRACTION RATE OF EXPANDED SKIN IN RABBIT. Chinese Journal of Reparative and Reconstructive Surgery, 2015, 29(12): 1541-1546. doi: 10.7507/1002-1892.20150329 Copy

1.	Nie C, Yang D, Xu J, et al. Locallty administered adipos-derived stem cells accelerate wound healing through differentiation and vasculogenesis. Cell Transplant, 2011, 20(2):205-216.
2.	闫迎军, 乔群, 孙家明, 等. 几丁糖预防硅凝胶乳房假体包膜挛缩的实验研究. 中华整形外科杂志, 2005, 21(2):139-142.
3.	施文斌, 董玉英, 董福生, 等. 不同扩张方式对扩张皮肤胶原纤维和肌动蛋白的影响. 实用口腔医学杂志, 2010, 26(1):38-42.
4.	李江, 鲁开化, 艾玉峰, 等. 扩张后皮肤软组织收缩机理的实验研究. 中华整形外科杂志, 2002, 18(3):173-174.
5.	李薇薇, 冯锐. 富血小板血浆凝胶制备及对大鼠皮瓣成活的作用分析. 中国修复重建外科杂志, 2012, 26(1):64-69.
6.	Özdemir B, Kurtiş B, Tüter G, et al. Double-application of plateletrich plasma on bone healing in rabbits. Med Oral Patol Oral Cir Bucal, 2012, 17(1):e171-177.
7.	Lee HW, Reddy MS, Geurs N, et al. Efficacy of platelet-rich plasma on wound healing in rabbits. J Periodontol, 2008, 79(4):691-696.
8.	Planat-Benard V, Silvestre JS, Cousin B, et al. Plasticity of human adipose lineage cells toward endothelial cells:physiological and therapeutic perspectives. Circulation, 2004, 109(5):656-663.
9.	郭垣杉, 颜玲. 无血清培养促进脂肪干细胞向血管内皮细胞分化. 中国组织工程研究, 2013, 17(36):6443-6448.
10.	Cao Y, Sun Z, Liao L, et al. Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo. Biochem Biophys Res Commun, 2005, 332(2):370-379.
11.	Deveza L, Choi J, Imanbayev G, et al. Paracrine release from nonviral engineered adipose-derived stem cells promotes endothelial cell survival and migration in vitro. Stem Cells Dev, 2012, 22(3):483-491.
12.	Francis AJ, Marks R. Skin stretching and epidermopoiesis. Br J Exp Pothol, 1977, 58(1):35-39.
13.	Marshall WR, Godfrey M, Hollister D, et al. Types of collagen in breast capsules. Ann Plast Surg, 1989, 23(5):401-405.
14.	刘学军, 郝建忠, 喻理. 皮肤软组织扩张术中维持扩张期对扩张皮肤的影响. 国际医药卫生导报, 2002, (4):48-49.
15.	杨宇. 扩张包膜血管构筑的试验研究及其临床应用. 北京:中国协和医科大学, 2006.
16.	鲁开化, 周庆红. 皮肤扩张术在美容整形外科应用的现状及未来. 中国实用美容整形外科杂志, 2005, 16(3):129-130.
17.	刘文阁, 杨佩瑛, 马桂娥, 等. 罂粟碱作用下扩张器周围纤维包膜超微结构的变化. 中华医学美容杂志, 1997, 3(4):178-181.
18.	范昌, 邢宏利, 李丽莎, 等. 透明质酸对扩张囊外纤维包膜影响的临床研究. 山西医药杂志, 2013, 42(5):504-506.
19.	刘文阁, 季莹, 杨佩瑛, 等. 曲安缩松加速组织扩张的临床应用研究. 中华整形烧伤外科杂志, 1999, 15(1):41-43.
20.	肖海军, 侯春林, 顾其胜, 等. 几丁糖膜预防术后粘连的研究进展. 上海生物医学工程, 2004, 25(3):47-52.
21.	仇树林, 闫焱, 张培培, 等. 几丁糖对扩张后皮瓣纤维包膜中毛细血管数量影响的实验研究. 中国美容整形外科杂志, 2008, 19(4):312-314.

1. Nie C, Yang D, Xu J, et al. Locallty administered adipos-derived stem cells accelerate wound healing through differentiation and vasculogenesis. Cell Transplant, 2011, 20(2):205-216.
2. 闫迎军, 乔群, 孙家明, 等. 几丁糖预防硅凝胶乳房假体包膜挛缩的实验研究. 中华整形外科杂志, 2005, 21(2):139-142.
3. 施文斌, 董玉英, 董福生, 等. 不同扩张方式对扩张皮肤胶原纤维和肌动蛋白的影响. 实用口腔医学杂志, 2010, 26(1):38-42.
4. 李江, 鲁开化, 艾玉峰, 等. 扩张后皮肤软组织收缩机理的实验研究. 中华整形外科杂志, 2002, 18(3):173-174.
5. 李薇薇, 冯锐. 富血小板血浆凝胶制备及对大鼠皮瓣成活的作用分析. 中国修复重建外科杂志, 2012, 26(1):64-69.
6. Özdemir B, Kurtiş B, Tüter G, et al. Double-application of plateletrich plasma on bone healing in rabbits. Med Oral Patol Oral Cir Bucal, 2012, 17(1):e171-177.
7. Lee HW, Reddy MS, Geurs N, et al. Efficacy of platelet-rich plasma on wound healing in rabbits. J Periodontol, 2008, 79(4):691-696.
8. Planat-Benard V, Silvestre JS, Cousin B, et al. Plasticity of human adipose lineage cells toward endothelial cells:physiological and therapeutic perspectives. Circulation, 2004, 109(5):656-663.
9. 郭垣杉, 颜玲. 无血清培养促进脂肪干细胞向血管内皮细胞分化. 中国组织工程研究, 2013, 17(36):6443-6448.
10. Cao Y, Sun Z, Liao L, et al. Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo. Biochem Biophys Res Commun, 2005, 332(2):370-379.
11. Deveza L, Choi J, Imanbayev G, et al. Paracrine release from nonviral engineered adipose-derived stem cells promotes endothelial cell survival and migration in vitro. Stem Cells Dev, 2012, 22(3):483-491.
12. Francis AJ, Marks R. Skin stretching and epidermopoiesis. Br J Exp Pothol, 1977, 58(1):35-39.
13. Marshall WR, Godfrey M, Hollister D, et al. Types of collagen in breast capsules. Ann Plast Surg, 1989, 23(5):401-405.
14. 刘学军, 郝建忠, 喻理. 皮肤软组织扩张术中维持扩张期对扩张皮肤的影响. 国际医药卫生导报, 2002, (4):48-49.
15. 杨宇. 扩张包膜血管构筑的试验研究及其临床应用. 北京:中国协和医科大学, 2006.
16. 鲁开化, 周庆红. 皮肤扩张术在美容整形外科应用的现状及未来. 中国实用美容整形外科杂志, 2005, 16(3):129-130.
17. 刘文阁, 杨佩瑛, 马桂娥, 等. 罂粟碱作用下扩张器周围纤维包膜超微结构的变化. 中华医学美容杂志, 1997, 3(4):178-181.
18. 范昌, 邢宏利, 李丽莎, 等. 透明质酸对扩张囊外纤维包膜影响的临床研究. 山西医药杂志, 2013, 42(5):504-506.
19. 刘文阁, 季莹, 杨佩瑛, 等. 曲安缩松加速组织扩张的临床应用研究. 中华整形烧伤外科杂志, 1999, 15(1):41-43.
20. 肖海军, 侯春林, 顾其胜, 等. 几丁糖膜预防术后粘连的研究进展. 上海生物医学工程, 2004, 25(3):47-52.
21. 仇树林, 闫焱, 张培培, 等. 几丁糖对扩张后皮瓣纤维包膜中毛细血管数量影响的实验研究. 中国美容整形外科杂志, 2008, 19(4):312-314.

Chinese Journal of Reparative and Reconstructive Surgery

EFFECT OF ADIPOSE-DERIVED STEM CELLS COMBINED WITH CHITOSAN ON IMMEDIATE RETRACTION RATE OF EXPANDED SKIN IN RABBIT

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