REACTION OF HUMAN FIBROBLASTS FROM DIFFERENT SITES TO THE MECHANICAL STRESS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

 WANGZhiguo ¹ , KUANGRuixia ¹ , XUQuanchen ² , CHENZhenyu ¹ , LIPeng ³

1. Department of Burn and Plastic Surgery, the Affiliated Hospital of Qingdao University, Qingdao Shandong, 266003, P. R. China;
2. Department of Stomatology, the Affiliated Hospital of Qingdao University, Qingdao Shandong, 266003, P. R. China;
3. Department of Plastic Surgery, Weifang People's Hospital;

Corresponding?author：

WANGZhiguo, Email: wzg_qd@126.com

Keywords：

Fibroblasts; Cyclic stretch; Normal skin; Hypertrophic scar

DOI：

10.7507/1002-1892.20150101

Video：

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Objective To explore the reaction of normal skin fibroblasts from different sites of human body to cyclic stretch. Methods The normal skin tissues from scapular upper back and medial side of upper arm of 3 patients were cultured in vitro. Fibroblasts of experimental group were loaded by cyclic stretch with 10% amplitude for 24, 36, and 48 hours respectively. Fibroblasts of control group were cultured without cyclic stretch. The morphologic changes were observed using inverted microscope. CCK-8 method was used to detect the proliferation of the fibroblasts. The expressions of integrin β₁ mRNA, p130Crk-associated substance (P130Cas) mRNA, transform growth factor β₁ (TGF-β₁) mRNA, and collagen type Ⅰ α1 chain (COL1A1) mRNA were detected by real-time quantitative PCR. The protein levels of collagen type Ⅰ and TGF-β₁ were detected by ELISA. Results The cultured cells showed a significantly increased cell proliferation ability, and apparent orientation after the applied strain. The proliferation activity, mRNA expression levels of integrin β₁, P130Cas, and TGF-β₁, protein levels of TGF-β₁ in back skin were significantly higher than those in arm skin (P<0.05) when the fibroblasts were loaded for 36 and 48 hours, but no significant difference between back skin and arm skin at 24 hours (P>0.05). There was no significant difference in mRNA expression level of COL1A1 and protein level of collagen type Ⅰ between back skin and arm skin at 24, 36, and 48 hours (P>0.05). There was no significant difference in all above indexes between back skin and arm skin in control group (P>0.05). Conclusion Fibroblasts from scapular upper back and medial side of upper arm display different reactions to cyclic stretch, which indicates that there exists site difference in the reactions of fibroblasts to cyclic stretch. It might be related with the incidence of hypertrophic scar in different sites of the body.

Citation： WANGZhiguo, KUANGRuixia, XUQuanchen, CHENZhenyu, LIPeng. REACTION OF HUMAN FIBROBLASTS FROM DIFFERENT SITES TO THE MECHANICAL STRESS. Chinese Journal of Reparative and Reconstructive Surgery, 2015, 29(4): 467-471. doi: 10.7507/1002-1892.20150101 Copy

1.	Mustoe TA, Cooter RD, Gold MH, et al. International clinical recommendations on scar management. Plast Reconstr Surg, 2002, 110(2):560-571.
2.	Yagmur C, Akaishi S, Ogawa R, et al. Mechanical receptor-related mechanisms in scar management:a review and hypothesis. Plast Reconstr Surg, 2010, 126(2):426-434.
3.	Derderian CA, Bastidas N, Lerman OZ, et al. Mechanical strain alters gene expression in an in vitro model of hypertrophic scarring. Ann Plast Surg, 2005, 55(1):69-75.
4.	Ogawa R. Keloid and hypertrophic scarring may result from a mechanoreceptor or mechanosensitive nociceptor disorder. Med Hypotheses, 2008, 71(4):493-500.
5.	Ogawa R. Mechanobiology of scarring. Wound Repair Regen, 2011, 19 Suppl 1:s2-9.
6.	Ohmori Y, Akaishi S, Ogawa R, et al. The analysisof keloid favorite site. The 4th Japan Scar Workshop, Tokyo, Japan, 2009.
7.	Parsons M, Kessler E, Laurent GJ, et al. Mechanical load enhances procollagen processing in dermal fibroblasts by regulating levels of procollagen C-proteinase. Exp Cell Res, 1999, 252(2):319-331.
8.	Balestrini JL, Billiar KL. Equibiaxial cyclic stretch stimulates fibroblasts to rapidly remodel fibrin. J Biomech, 2006, 39(16):2983-2990.
9.	王志國, 匡瑞霞, 陳振雨, 等. 不同幅度牽張力對正常皮膚成纖維細胞向增生性瘢痕成纖維細胞轉化的誘導作用. 中華醫學雜志, 2015, 95(4):294-298.
10.	舒茂國, 易成剛, 韓巖, 等. 機械應力刺激對培養的人成纖維細胞分泌生長因子的影響. 中國美容醫學, 2008, 17(5):689-691.
11.	Syedain ZH, Tranquillo RT. TGF-β1 diminishes collagen production during long-term cyclic stretching of engineered connective tissue:implication of decreased ERK signaling. J Biomech, 2011, 44(5):848-855.
12.	Wipff PJ, Rifkin DB, Meister JJ, et al. Myofibroblast contraction activates latent TGF-beta1 from the extracellular matrix. J Cell Biol, 2007, 179(6):1311-1323.
13.	Wang Z, Fong KD, Phan TT, et al. Increased transcriptional response to mechanical strain in keloid fibroblasts due to increased focal adhesion complex formation. J Cell Physiol, 2006, 206(2):510-517.
14.	Kanazawa Y, Nomura J, Yoshimoto S, et al. Cyclical cell stretching of skin-derived fibroblasts downregulates connective tissue growth factor (CTGF) production. Connect Tissue Res, 2009, 50(5):323-329.
15.	Baker EL, Zaman MH. The biomechanical integrin. J Biomech, 2010, 43(1):38-44.
16.	Goldmann WH. Mechanotransduction and focal adhesions. Cell Biol Int, 2012, 36(7):649-652.
17.	Cheng M, Guan X, Li H, et al. Shear stress regulates late EPC differentiation via mechanosensitive molecule-mediated cytoskeletal rearrangement. PLoS One, 2013, 8(7):e67675.
18.	Wang Y, McNiven MA. Invasive matrix degradation at focal adhesions occurs via protease recruitment by a FAK-p130Cas complex. J Cell Biol, 2012, 196(3):375-385.
19.	Jano?tiak R, Brábek J, Auernheimer V, et al. CAS directly interacts with vinculin to control mechanosensing and focal adhesion dynamics. Cell Mol Life Sci, 2014, 71(4):727-744.
20.	Hannafin JA, Attia EA, Henshaw R, et al. Effect of cyclic strain and plating matrix on cell proliferation and integrin expression by ligament fibroblasts. J Orthop Res, 2006, 24(2):149-158.
21.	Demou ZN. Gene expression profiles in 3D tumor analogs indicate compressive strain differentially enhances metastatic potential. Ann Biomed Eng, 2010, 38(11):3509-3520.

1. Mustoe TA, Cooter RD, Gold MH, et al. International clinical recommendations on scar management. Plast Reconstr Surg, 2002, 110(2):560-571.
2. Yagmur C, Akaishi S, Ogawa R, et al. Mechanical receptor-related mechanisms in scar management:a review and hypothesis. Plast Reconstr Surg, 2010, 126(2):426-434.
3. Derderian CA, Bastidas N, Lerman OZ, et al. Mechanical strain alters gene expression in an in vitro model of hypertrophic scarring. Ann Plast Surg, 2005, 55(1):69-75.
4. Ogawa R. Keloid and hypertrophic scarring may result from a mechanoreceptor or mechanosensitive nociceptor disorder. Med Hypotheses, 2008, 71(4):493-500.
5. Ogawa R. Mechanobiology of scarring. Wound Repair Regen, 2011, 19 Suppl 1:s2-9.
6. Ohmori Y, Akaishi S, Ogawa R, et al. The analysisof keloid favorite site. The 4th Japan Scar Workshop, Tokyo, Japan, 2009.
7. Parsons M, Kessler E, Laurent GJ, et al. Mechanical load enhances procollagen processing in dermal fibroblasts by regulating levels of procollagen C-proteinase. Exp Cell Res, 1999, 252(2):319-331.
8. Balestrini JL, Billiar KL. Equibiaxial cyclic stretch stimulates fibroblasts to rapidly remodel fibrin. J Biomech, 2006, 39(16):2983-2990.
9. 王志國, 匡瑞霞, 陳振雨, 等. 不同幅度牽張力對正常皮膚成纖維細胞向增生性瘢痕成纖維細胞轉化的誘導作用. 中華醫學雜志, 2015, 95(4):294-298.
10. 舒茂國, 易成剛, 韓巖, 等. 機械應力刺激對培養的人成纖維細胞分泌生長因子的影響. 中國美容醫學, 2008, 17(5):689-691.
11. Syedain ZH, Tranquillo RT. TGF-β1 diminishes collagen production during long-term cyclic stretching of engineered connective tissue:implication of decreased ERK signaling. J Biomech, 2011, 44(5):848-855.
12. Wipff PJ, Rifkin DB, Meister JJ, et al. Myofibroblast contraction activates latent TGF-beta1 from the extracellular matrix. J Cell Biol, 2007, 179(6):1311-1323.
13. Wang Z, Fong KD, Phan TT, et al. Increased transcriptional response to mechanical strain in keloid fibroblasts due to increased focal adhesion complex formation. J Cell Physiol, 2006, 206(2):510-517.
14. Kanazawa Y, Nomura J, Yoshimoto S, et al. Cyclical cell stretching of skin-derived fibroblasts downregulates connective tissue growth factor (CTGF) production. Connect Tissue Res, 2009, 50(5):323-329.
15. Baker EL, Zaman MH. The biomechanical integrin. J Biomech, 2010, 43(1):38-44.
16. Goldmann WH. Mechanotransduction and focal adhesions. Cell Biol Int, 2012, 36(7):649-652.
17. Cheng M, Guan X, Li H, et al. Shear stress regulates late EPC differentiation via mechanosensitive molecule-mediated cytoskeletal rearrangement. PLoS One, 2013, 8(7):e67675.
18. Wang Y, McNiven MA. Invasive matrix degradation at focal adhesions occurs via protease recruitment by a FAK-p130Cas complex. J Cell Biol, 2012, 196(3):375-385.
19. Jano?tiak R, Brábek J, Auernheimer V, et al. CAS directly interacts with vinculin to control mechanosensing and focal adhesion dynamics. Cell Mol Life Sci, 2014, 71(4):727-744.
20. Hannafin JA, Attia EA, Henshaw R, et al. Effect of cyclic strain and plating matrix on cell proliferation and integrin expression by ligament fibroblasts. J Orthop Res, 2006, 24(2):149-158.
21. Demou ZN. Gene expression profiles in 3D tumor analogs indicate compressive strain differentially enhances metastatic potential. Ann Biomed Eng, 2010, 38(11):3509-3520.

Chinese Journal of Reparative and Reconstructive Surgery

REACTION OF HUMAN FIBROBLASTS FROM DIFFERENT SITES TO THE MECHANICAL STRESS

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