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.	閆迎軍, 喬群, 孫家明, 等. 幾丁糖預防硅凝膠乳房假體包膜攣縮的實驗研究. 中華整形外科雜志, 2005, 21(2):139-142.
3.	施文斌, 董玉英, 董福生, 等. 不同擴張方式對擴張皮膚膠原纖維和肌動蛋白的影響. 實用口腔醫學雜志, 2010, 26(1):38-42.
4.	李江, 魯開化, 艾玉峰, 等. 擴張后皮膚軟組織收縮機理的實驗研究. 中華整形外科雜志, 2002, 18(3):173-174.
5.	李薇薇, 馮銳. 富血小板血漿凝膠制備及對大鼠皮瓣成活的作用分析. 中國修復重建外科雜志, 2012, 26(1):64-69.
6.	?zdemir B, Kurti? 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, 顏玲. 無血清培養促進脂肪干細胞向血管內皮細胞分化. 中國組織工程研究, 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.	劉學軍, 郝建忠, 喻理. 皮膚軟組織擴張術中維持擴張期對擴張皮膚的影響. 國際醫藥衛生導報, 2002, (4):48-49.
15.	楊宇. 擴張包膜血管構筑的試驗研究及其臨床應用. 北京:中國協和醫科大學, 2006.
16.	魯開化, 周慶紅. 皮膚擴張術在美容整形外科應用的現狀及未來. 中國實用美容整形外科雜志, 2005, 16(3):129-130.
17.	劉文閣, 楊佩瑛, 馬桂娥, 等. 罌粟堿作用下擴張器周圍纖維包膜超微結構的變化. 中華醫學美容雜志, 1997, 3(4):178-181.
18.	范昌, 邢宏利, 李麗莎, 等. 透明質酸對擴張囊外纖維包膜影響的臨床研究. 山西醫藥雜志, 2013, 42(5):504-506.
19.	劉文閣, 季瑩, 楊佩瑛, 等. 曲安縮松加速組織擴張的臨床應用研究. 中華整形燒傷外科雜志, 1999, 15(1):41-43.
20.	肖海軍, 侯春林, 顧其勝, 等. 幾丁糖膜預防術后粘連的研究進展. 上海生物醫學工程, 2004, 25(3):47-52.
21.	仇樹林, 閆焱, 張培培, 等. 幾丁糖對擴張后皮瓣纖維包膜中毛細血管數量影響的實驗研究. 中國美容整形外科雜志, 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. 閆迎軍, 喬群, 孫家明, 等. 幾丁糖預防硅凝膠乳房假體包膜攣縮的實驗研究. 中華整形外科雜志, 2005, 21(2):139-142.
3. 施文斌, 董玉英, 董福生, 等. 不同擴張方式對擴張皮膚膠原纖維和肌動蛋白的影響. 實用口腔醫學雜志, 2010, 26(1):38-42.
4. 李江, 魯開化, 艾玉峰, 等. 擴張后皮膚軟組織收縮機理的實驗研究. 中華整形外科雜志, 2002, 18(3):173-174.
5. 李薇薇, 馮銳. 富血小板血漿凝膠制備及對大鼠皮瓣成活的作用分析. 中國修復重建外科雜志, 2012, 26(1):64-69.
6. ?zdemir B, Kurti? 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, 顏玲. 無血清培養促進脂肪干細胞向血管內皮細胞分化. 中國組織工程研究, 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. 劉學軍, 郝建忠, 喻理. 皮膚軟組織擴張術中維持擴張期對擴張皮膚的影響. 國際醫藥衛生導報, 2002, (4):48-49.
15. 楊宇. 擴張包膜血管構筑的試驗研究及其臨床應用. 北京:中國協和醫科大學, 2006.
16. 魯開化, 周慶紅. 皮膚擴張術在美容整形外科應用的現狀及未來. 中國實用美容整形外科雜志, 2005, 16(3):129-130.
17. 劉文閣, 楊佩瑛, 馬桂娥, 等. 罌粟堿作用下擴張器周圍纖維包膜超微結構的變化. 中華醫學美容雜志, 1997, 3(4):178-181.
18. 范昌, 邢宏利, 李麗莎, 等. 透明質酸對擴張囊外纖維包膜影響的臨床研究. 山西醫藥雜志, 2013, 42(5):504-506.
19. 劉文閣, 季瑩, 楊佩瑛, 等. 曲安縮松加速組織擴張的臨床應用研究. 中華整形燒傷外科雜志, 1999, 15(1):41-43.
20. 肖海軍, 侯春林, 顧其勝, 等. 幾丁糖膜預防術后粘連的研究進展. 上海生物醫學工程, 2004, 25(3):47-52.
21. 仇樹林, 閆焱, 張培培, 等. 幾丁糖對擴張后皮瓣纖維包膜中毛細血管數量影響的實驗研究. 中國美容整形外科雜志, 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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