An Investigation of Antibacterial Pretreatment Protocol for Primary Fibroblast Culture from Transbronchial Biopsies in Benign Tracheal Stenosis Patients_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

WangYingzhi , ChenYu ,  LiShiyue

National Clinical Research Center for Respiratory Disease, State Key Lab of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510120, China;

Corresponding?author：

LiShiyue, Email: lishiyue@188.com

Keywords：

Granulation tissue; Fibroblasts; Primary cell culture; Antibiotics; Bacteria

DOI：

10.7507/1671-6205.2015070

Video：

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Objective To investigate the antibacterial pretreatment protocol for primary fibroblast cell culture from transbronchial biopsies in patients with benign tracheal stenosis (BTS). Methods Fifteen specimens of intratracheal hyperplastic granulation tissue were obtained from 14 BTS patients by transbronchial biopsies. The tissues were divided into 3 groups according to different antibacterial pretreatment with 5 specimens in each group. An usual concentration of antibiotics pretreatment group (group 1) was pretreated by washing with PBS contained 1%-2% penicillin/streptomycin. A high concentration of antibiotics pretreatment group (group 2) was pretreated by washing with PBS contained 6% penicillin/streptomycin. An alchohol and high concentration of antibiotics pretreatment group (group 3) was pretreated by washing with 75% alcohol 3-4 seconds firstly,then by washing with 6% penicillin/streptomycin. After different pretreatment,all tissues were cultured with tissue culture method in the same condition. Results The primary fibroblasts were successfully cultured from the tissue pretreated by method 2 and 3,but not cultured from the tissue pretreated by method 1 with large amount of bacteria. There were significant differences in the furthest radius of cell proliferation between different culture time points in three groups (P<0.01). The differences in the furthest radius of cell proliferation between three groups were not different at 24,48 or 72 h (P>0.05),but were significant between three groups at 96 h (P<0.05). Conclusion An pretreatment protocol with high concentration of antibiotics or 75% alcohol is feasible in human primary fibroblasts culture from small specimens obtained by transbronchial biopsy.

Citation： WangYingzhi, ChenYu, LiShiyue. An Investigation of Antibacterial Pretreatment Protocol for Primary Fibroblast Culture from Transbronchial Biopsies in Benign Tracheal Stenosis Patients. Chinese Journal of Respiratory and Critical Care Medicine, 2015, 14(3): 286-290. doi: 10.7507/1671-6205.2015070 Copy

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11.	章伏生,陸樹良.肌成纖維細胞在皮膚瘢痕形成過程中的作用.中華創傷雜志,2009,25:1106-1109.
12.	Singh T, Sandulache VC, Otteson TD,et al.Subglottic stenosis examined as a fibrotic response to airway injury characterized by altered mucosal fibroblast activity. Arch Otolaryngol Head Neck Surg, 2010,136:163-170.
13.	陳楠,張杰,徐敏,等.人氣道肉芽組織成纖維細胞的原代培養方法.中華結核和呼吸雜志,2013,36:280-282.
14.	馬迎民,潘亮,謝菲,等.抗生素的預防應用與重癥患者氣道內寄生菌的分布.中華醫院感染學雜志,2005,15:70-72.

1. ?楊希之,毛小慧,敖華飛,等.5-氟尿嘧啶局部治療兔瘢痕性喉氣管狹窄的實驗研究.中國耳鼻咽喉顱底外科雜志,2011,03:175-180.
2. Guven M, Turan F, Eyibilen A, et al. A comparison of the efficacy of 5-fluorouracil/triamcinolone, carnitine and dexamethasone therapy on wound healing in tracheal injury: potential for preventing tracheal stenosis? Eur Arch Otorhinolaryngol,2012, 269:201-206.
3. Koc S, Kiyici H, S?A?Güt E, et al. Effect of pentoxifylline and 5-fluorouracil/triamcinolone on laryngotracheal stenosis developing as a complication of tracheostomy: study in rats. Eur Arch Otorhinolaryngol,2012, 269:1813-1820.
4. 鮑榮輝,劉先哲,周軍.人成纖維細胞的原代培養及鑒定.新鄉醫學院學報,2011,5:577-579.
5. 張卓然,主編.培養細胞學與細胞培養技術.上海;上海科學技術出版社,2004,182-206.
6. 李時悅.良性中央氣道狹窄的介入治療方法選擇.中華結核和呼吸雜志,2011,34:329-332.
7. Rahman NA, Fruchter O, Shitrit D,et al. Flexible bronchoscopic management of benign tracheal stenosis: long term follow-up of 115 patients. J Cardiothorac Surg, 2010, 5:2.
8. Thornton RH, Gordon RL, Kerlan RK, et al. Outcomes of tracheobronchial stent placement for benign disease. Radiology,2006,240: 273-282.
9. Cosano PA, Munoz CL, Cosano PFJ, et al.Endoscopic treatment of central airway stenosis: five years' experience. Arch Bronconeumol,2005,41: 322-327.
10. 蘇柱泉,李時悅. 良性氣道狹窄治療后再狹窄的原因分析與預防.中華臨床醫師雜志(電子版),2013,7:3122-3125.
11. 章伏生,陸樹良.肌成纖維細胞在皮膚瘢痕形成過程中的作用.中華創傷雜志,2009,25:1106-1109.
12. Singh T, Sandulache VC, Otteson TD,et al.Subglottic stenosis examined as a fibrotic response to airway injury characterized by altered mucosal fibroblast activity. Arch Otolaryngol Head Neck Surg, 2010,136:163-170.
13. 陳楠,張杰,徐敏,等.人氣道肉芽組織成纖維細胞的原代培養方法.中華結核和呼吸雜志,2013,36:280-282.
14. 馬迎民,潘亮,謝菲,等.抗生素的預防應用與重癥患者氣道內寄生菌的分布.中華醫院感染學雜志,2005,15:70-72.

Chinese Journal of Respiratory and Critical Care Medicine

An Investigation of Antibacterial Pretreatment Protocol for Primary Fibroblast Culture from Transbronchial Biopsies in Benign Tracheal Stenosis Patients

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