A rapid histological preparation method for observation of morphology and composition distribution of tendon collagen fascicle and endotendinium_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Yi ¹ , ZHOU Li ¹ , TANG Linqiao ¹ , QIN Tingwu ² ,  NING Liangju ²

1. Research Core Facility of West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China;
2. Laboratory of Stem Cell and Tissue Engineering, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China;

Corresponding?author：

NING Liangju, Email: nlj1024@163.com

Keywords：

Tendon; frozen section; baking; Masson staining

DOI：

10.7507/1002-1892.201903101

Video：

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Objective To explore a rapid histological preparation method to observe morphology and composition distribution of tendon collagen fascicle and endotendinum. Methods Taking porcine superflexor tendon of foot as an example, tendons were sliced into sections with 6 μm by frozen section technology, after which general observation of the section integrity was carried out. After fixed with 10% neutral buffered formalin and performed with HE staining, the tissue integrity and ice crystal formation were observed under microscope. Sections were then divided into 5 groups by different methods of dyeing. Group A: Priodic acid-Shiff (PAS) staining; group B: Masson staining; group C: reticular fibers staining; group D: immunohistochemical and immunofluorescent staining of type Ⅲ collagen; group E: the sections were baked at 65℃ for 10 minutes and stained with Masson. The composition distribution of tendon collagen fascicle and endotendinum in different groups were observed. Results From general observation, the frozen section of tendon tissue was complete and continuous. Although the tissue integrity in the tendon sections could be seen and no ice crystal was formed, the composition distribution could not be identified by HE staining. The entire tendons in groups A, B, and C were dyed, and the composition distribution of collagen fascicle and endotendinum could not be identified. The endotendinum in group D was stained weakly positive for type Ⅲ collagen alone, and the two components were differentiated dyed but the contrast was not obvious. In group E, the collagen fascicle and endotendinium were differentiated dyed and the two components in tendon tissue were clearly visible. Conclusion The morphology and the composition distribution of tendon collagen fascicle and endotendinum can be characterized rapidly and accurately, using a combination of baking at 65℃ for 10 minutes and Masson staining after porcine superflexor tendons were sliced by frozen section technology.

Citation： ZHANG Yi, ZHOU Li, TANG Linqiao, QIN Tingwu, NING Liangju. A rapid histological preparation method for observation of morphology and composition distribution of tendon collagen fascicle and endotendinium. Chinese Journal of Reparative and Reconstructive Surgery, 2019, 33(9): 1169-1173. doi: 10.7507/1002-1892.201903101 Copy

1.	Liu Y, Ramanath HS, Wang DA. Tendon tissue engineering using scaffold enhancing strategies. Trends Biotechnol, 2008, 26(4): 201-209.
2.	Kannus P. Structure of the tendon connective tissue. Scand J Med Sci Sports, 2000, 10(6): 312-320.
3.	Qin TW, Chen Q, Sun YL, et al. Mechanical characteristics of native tendon slices for tissue engineering scaffold. J Biomed Mater Res B Appl Biomater, 2012, 100B(3): 752-758.
4.	顧怡瑾, 陸新元, 付華輝, 等. Masson 染色在輔助診斷肝纖維化中應用. 臨床與實驗病理學雜志, 2012, 28(1): 101-102.
5.	楊小榮, 劉芳, 汪芳, 等. Masson 染色法在周圍神經損傷中的應用. 診斷病理學雜志, 2012, 19(6): 473-474.
6.	Huang YZ, Wang JJ, Huang YC, et al. Organic composite-mediated surface coating of human acellular bone matrix with strontium. Mater Sci Eng C Mater Biol Appl, 2017, 84: 12-20.
7.	Ni P, Ding Q, Fan M, et al. Injectable thermosensitive PEG-PCL-PEG hydrogel/acellular bone matrix composite for bone regeneration in cranial defects. Biomaterials, 2014, 35(1): 236-248.
8.	Wang ZL, Wu SZ, Li ZF, et al. Comparison of small intestinal submucosa and polypropylene mesh for abdominal wall defect repair. J Biomater Sci Polym Ed, 2018, 29(6): 1-20.
9.	He SK, Guo JH, Wang ZL, et al. Efficacy and safety of small intestinal submucosa in dural defect repair in a canine model. Mater Sci Eng C Mater Biol Appl, 2017, 73: 267-274.
10.	韓天龍, 王敏, 紅敏, 等. Masson 三色染色與 HE 染色在豚鼠牙齒組織石蠟切片中的比較研究及應用. 中國畜牧獸醫, 2011, 38(3): 55-57.
11.	王玨, 朱禮國, 唐幕湘, 等. 改良 Masson 三色染色法在膠原纖維中的應用. 鄖陽醫學院學報, 2006, 25(1): 44.
12.	張哲, 王偉, 商建峰, 等. 膠原纖維特殊染色法在心肌纖維化診斷中的應用比較. 診斷病理學雜志, 2016, 23(8): 578-580.
13.	蔡新華, 郭志坤, 霍殿樣. 石蠟包埋對兔肝、腎組織塊的收縮影響. 解剖科學進展, 2000, 6(4): 380-381.
14.	蔡新華, 郭志坤. 石蠟包埋對心、肺和脾組織的收縮影響. 解剖學雜志, 2002, 25(2): 198-199.
15.	王華, 羅殿中. 術中冰凍切片制片的規范化操作與常見問題原因分析. 2011 年全國西安病理技術學術會議暨全軍病理技術學術會議論文集. 西安: 解放軍醫學科學技術委員會, 2011.
16.	包翠芬, 劉霞, 穆長征, 等. 比較三種防冰晶法對切片保存效果的影響. 中國組織化學與細胞化學雜志, 2007, 16(1): 122-123.
17.	楊秀靜, 滕孝靜. 冰凍切片技術經驗探討. 臨床和實驗醫學雜志, 2014, 13(23): 2002-2004.
18.	王玨, 朱禮國, 張健, 等. 腎臟 PAS 染色法的探討. 實用醫技雜志, 2006, 13(22): 4097.
19.	鐘妮娜, 耿毅, 彭西. PAS 染色技術的改進. 解剖學雜志, 2005, 28(6): 723-724.
20.	胡沛臻, 馬福成, 張傳山, 等. 微波網狀纖維染色法在病理診斷中的應用. 診斷病理學雜志, 2002, 9(4): 249.
21.	李萍萍, 葛青. 軟骨寡聚基質蛋白 (COMP) 對 B 淋巴細胞發育及功能的影響.中國細胞生物學學會全國學術大會論文集. 武漢: 中國細胞生物學學會, 2013.
22.	魏壯, 崔樹森, 張巨, 等. 預防肌腱粘連的研究與進展. 吉林大學學報, 2001, 27(2): 218-220.
23.	Franchi M, Trirè A, Quaranta M, et al. Collagen structure of tendon relates to function. The Scientific World Journal, 2017, 7: 404-420.
24.	解立武, 常芳, 盧小波. 經典和改良 Gomori 網狀纖維染色的比較. 中國藥物與臨床, 2018, 18(1): 31-32.
25.	蔡新華, 郭志坤, 毛會麗, 等. 大鼠心肌組織中網狀纖維組織學構筑及增齡變化. 解剖學報, 2009, 40(1): 127-129.
26.	廖大清, 馬玉瓊, 陳思秀. 一種顯示網狀纖維的方法. 四川解剖學雜志, 2001, 9(2): 92-93.
27.	韓慶偉, 張云. 特殊染色網狀纖維在病理診斷中的應用. 實用醫技雜志, 2007, 14(11): 1420.
28.	田玉旺, 李琳, 朱紅艷, 等. 提高網狀纖維染色質量的技術探討. 中國組織化學與細胞學雜志, 2012, 21(3): 316-318.
29.	Wu F, Nerlich M, Docheva D. Tendon injuries: Basic science and repair proposals. EFFORT Open Rev, 2017, 2(7): 332-342.

1. Liu Y, Ramanath HS, Wang DA. Tendon tissue engineering using scaffold enhancing strategies. Trends Biotechnol, 2008, 26(4): 201-209.
2. Kannus P. Structure of the tendon connective tissue. Scand J Med Sci Sports, 2000, 10(6): 312-320.
3. Qin TW, Chen Q, Sun YL, et al. Mechanical characteristics of native tendon slices for tissue engineering scaffold. J Biomed Mater Res B Appl Biomater, 2012, 100B(3): 752-758.
4. 顧怡瑾, 陸新元, 付華輝, 等. Masson 染色在輔助診斷肝纖維化中應用. 臨床與實驗病理學雜志, 2012, 28(1): 101-102.
5. 楊小榮, 劉芳, 汪芳, 等. Masson 染色法在周圍神經損傷中的應用. 診斷病理學雜志, 2012, 19(6): 473-474.
6. Huang YZ, Wang JJ, Huang YC, et al. Organic composite-mediated surface coating of human acellular bone matrix with strontium. Mater Sci Eng C Mater Biol Appl, 2017, 84: 12-20.
7. Ni P, Ding Q, Fan M, et al. Injectable thermosensitive PEG-PCL-PEG hydrogel/acellular bone matrix composite for bone regeneration in cranial defects. Biomaterials, 2014, 35(1): 236-248.
8. Wang ZL, Wu SZ, Li ZF, et al. Comparison of small intestinal submucosa and polypropylene mesh for abdominal wall defect repair. J Biomater Sci Polym Ed, 2018, 29(6): 1-20.
9. He SK, Guo JH, Wang ZL, et al. Efficacy and safety of small intestinal submucosa in dural defect repair in a canine model. Mater Sci Eng C Mater Biol Appl, 2017, 73: 267-274.
10. 韓天龍, 王敏, 紅敏, 等. Masson 三色染色與 HE 染色在豚鼠牙齒組織石蠟切片中的比較研究及應用. 中國畜牧獸醫, 2011, 38(3): 55-57.
11. 王玨, 朱禮國, 唐幕湘, 等. 改良 Masson 三色染色法在膠原纖維中的應用. 鄖陽醫學院學報, 2006, 25(1): 44.
12. 張哲, 王偉, 商建峰, 等. 膠原纖維特殊染色法在心肌纖維化診斷中的應用比較. 診斷病理學雜志, 2016, 23(8): 578-580.
13. 蔡新華, 郭志坤, 霍殿樣. 石蠟包埋對兔肝、腎組織塊的收縮影響. 解剖科學進展, 2000, 6(4): 380-381.
14. 蔡新華, 郭志坤. 石蠟包埋對心、肺和脾組織的收縮影響. 解剖學雜志, 2002, 25(2): 198-199.
15. 王華, 羅殿中. 術中冰凍切片制片的規范化操作與常見問題原因分析. 2011 年全國西安病理技術學術會議暨全軍病理技術學術會議論文集. 西安: 解放軍醫學科學技術委員會, 2011.
16. 包翠芬, 劉霞, 穆長征, 等. 比較三種防冰晶法對切片保存效果的影響. 中國組織化學與細胞化學雜志, 2007, 16(1): 122-123.
17. 楊秀靜, 滕孝靜. 冰凍切片技術經驗探討. 臨床和實驗醫學雜志, 2014, 13(23): 2002-2004.
18. 王玨, 朱禮國, 張健, 等. 腎臟 PAS 染色法的探討. 實用醫技雜志, 2006, 13(22): 4097.
19. 鐘妮娜, 耿毅, 彭西. PAS 染色技術的改進. 解剖學雜志, 2005, 28(6): 723-724.
20. 胡沛臻, 馬福成, 張傳山, 等. 微波網狀纖維染色法在病理診斷中的應用. 診斷病理學雜志, 2002, 9(4): 249.
21. 李萍萍, 葛青. 軟骨寡聚基質蛋白 (COMP) 對 B 淋巴細胞發育及功能的影響.中國細胞生物學學會全國學術大會論文集. 武漢: 中國細胞生物學學會, 2013.
22. 魏壯, 崔樹森, 張巨, 等. 預防肌腱粘連的研究與進展. 吉林大學學報, 2001, 27(2): 218-220.
23. Franchi M, Trirè A, Quaranta M, et al. Collagen structure of tendon relates to function. The Scientific World Journal, 2017, 7: 404-420.
24. 解立武, 常芳, 盧小波. 經典和改良 Gomori 網狀纖維染色的比較. 中國藥物與臨床, 2018, 18(1): 31-32.
25. 蔡新華, 郭志坤, 毛會麗, 等. 大鼠心肌組織中網狀纖維組織學構筑及增齡變化. 解剖學報, 2009, 40(1): 127-129.
26. 廖大清, 馬玉瓊, 陳思秀. 一種顯示網狀纖維的方法. 四川解剖學雜志, 2001, 9(2): 92-93.
27. 韓慶偉, 張云. 特殊染色網狀纖維在病理診斷中的應用. 實用醫技雜志, 2007, 14(11): 1420.
28. 田玉旺, 李琳, 朱紅艷, 等. 提高網狀纖維染色質量的技術探討. 中國組織化學與細胞學雜志, 2012, 21(3): 316-318.
29. Wu F, Nerlich M, Docheva D. Tendon injuries: Basic science and repair proposals. EFFORT Open Rev, 2017, 2(7): 332-342.

Chinese Journal of Reparative and Reconstructive Surgery

A rapid histological preparation method for observation of morphology and composition distribution of tendon collagen fascicle and endotendinium

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