Research Article
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Is Enoxaparin Sodium Exactly Safe For Subcutaneous Fibroblast?: A Cell Culture Study

Year 2021, Volume 5, Issue 1, 18 - 23, 23.04.2021
https://doi.org/10.30565/medalanya.822679

Abstract

Aim: Despite relatively low amount in the subcutaneous tissue, fibroblasts play a critical role in the continuity of intercellular connections, maintenance of tissue integrity, and forming a balanced fascial network. Enoxaparin sodium is widely used in the prophylaxis and treatment of deep vein thrombosis. In the present study, we aimed to examine the cytotoxic and apoptotic effects of enoxaparin sodium on fibroblast cells in an in vitro model.

Material and Methods: In a mouse model, L929 fibroblast cells were treated with enoxaparin sodium 4000 IU, 2000 IU, 1000 IU, 500 IU, and 250 IU. At 48 hours, cell morphology was evaluated; cell viability was analyzed through methylthiazole tetrazolium assay and apoptosis was assessed by propidium iodide/ acridine orange staining.

Results: The test results showed that high doses (4000 IU, 2000 IU) exerted cytotoxic effects and induced apoptotic morphology. Compared to the control group, there was no significant difference in the cell viability in Dilutions III, IV, and V.

Conclusion: Based on our results, despite prophylactic dose in the in vitro setting, high-dose enoxaparin showed cytotoxic and apoptotic effects. Long-term high-dose enoxaparin sodium may affect the number of subcutaneous fibroblasts, impairing the skin integrity and subcutaneous tissue healing

References

  • 1. Canales JF, Ferguson JJ. Low-Molecular-Weight Heparins Mechanisms, Trials,and Role in Contempo-rary Interventional Medicine. Am J Cardiovasc Drugs. 2008;8:15-25. PMID: 18303934
  • 2. Laurent P, Dussarat GV, Bonal J, Jego C, Talard P, Bouchiat C,et al. Low Molecular Weight Heparins A Guide to Their Optimum Use In Pregnancy. Drugs. 2002;62:463-77. PMID: 11827560
  • 3. Linkins LA. Heparin induced thrombocytopenia. BMJ. 2015;350:g7566. PMID: 25569604
  • 4. Atiq F, van den Bemt PM, Leebeek FW, van Gelder T, Versmissen J. A systematic review on the ac-cumulation of prophylactic dosages of low-molecular-weight heparins (LMWHs) in patients with renal insufficiency. Eur J Clin Pharmacol. 2015;71(8):921-9. PMID: 26071276
  • 5. An I, Harman M, Ibiloglu I, Bullous hemorrhagic dermatosis induced by enoxaparin. Indian Dermatol Online J. 2017;8(5):347-9. PMID: 28979868
  • 6. Bircher AJ, Harr T, Hohenstein L, Tsakiris DA. Hypersensitivity reactions to anticoagulant drugs: diag-nosis and management options. Allergy. 2006;61(12):1432-40. PMID: 17073874
  • 7. Ronceray S, Dinulescua M, Le Gall F, Polardc E, Dupuya A, Adamskia H. Enoxaparin- Induced DRESS Syndrome. Case Rep Dermatol. 2012;4(3):233–7. PMID:23185158
  • 8. Langevin HM, Cornbrooks CJ, Taatjes DJ. Fibroblasts form a body-wide cellular network. Histochem Cell Biol. 2004;122(1):7-15. PMID: 15221410
  • 9. Langevin HM, Bouffard NA, Churchill DL, Badger GJ. Connective tissue fibroblast response to acu-puncture: dose-dependent effect of bidirectional needle rotation. J Altern Complement Med. 2007;13(3):355-60. PMID: 17480137
  • 10. Fernandes IR, Russo FB, Pignatari GC, Evangelinellis MM, Tavolari S, Muotri AR ,et al. Fibroblast sources: Where can we get them?. Cytotechnology. 2016;68(2):223-8. PMID: 25060709
  • 11. Available at: https://www.anatomyatlases.org/MicroscopicAnatomy/Section03/ Plate0327.shtml Access date: 17.12.2019
  • 12. Gürpinar ÖA, Kubat E, Onur MA. Cytotoxic Activity of Apixaban on HeLa Cells: An in vitro Study. Hacettepe J Biol Chem. 2018;46(3):395–402. doi: 10.15671/HJBC.2018.247
  • 13. Kubat E, Gurpinar, Karasoy D, Onur MA. A link between cytotoxicity in cell culture and gastrointestinal side effects of oral anticoagulants: bench-to-bedside. Bratisl Med J. 2018;119(11):706-712. PMID: 30686004
  • 14. Hussain H, Raj LS., Ahmad S, Razak MFA, Mohamud WNW, Bakar J, et al. Determination of cell via-bility using acridine orange/propidium iodide dual-spectrofluorometry assay. Cogent Food & Agricul-ture. 2019:5:1582398:1-9. doi:10.1080/23311932.2019.1582398
  • 15. Ziani L, Chouaib S, Thiery J. Alteration of the Antitumor Immune Response by Cancer-Associated Fibroblasts. Front Immunol. 2018;9:414:1-14. PMID: 29545811
  • 16. Li B, Wang JH. Fibroblasts and myofibroblasts in wound healing: force generation and measurement. J Tissue Viability. 2011;20(4):108-20. PMID: 19995679
  • 17. Rippa AL, Kalabusheva EP, Vorotelyak EA. Regeneration of Dermis: Scarring and Cells Involved. Cells. 2019;8(6):607:1-30. PMID: 31216669
  • 18. Kobbi Z, Kraiem H, Benlasfar Z, Marouani A, Massoud T, Boubaker S, et al. Comparative subcutane-ous repeated toxicity study of enoxaparin products in rats. Regul Toxicol Pharmacol. 2017;84:9-17. PMID: 27965129
  • 19. Pföhler C, Müller CS, Pindur G, Eichler H, Schäfers HJ, Grundmann U, et al. Delayed-Type Heparin Allergy: Diagnostic Procedures and Treatment Alternatives- A Case Series Including 15 Patients. WAO Journal. 2008;1(12):194-9. PMID: 23282847
  • 20. Schindewolf M, Schwaner S, Wolter M, Kroll H, Recke A, Kaufmann R, et al. Incidence and causes of heparin-induced skin lesions. CMAJ. 2009;181(8):477-81. PMID: 19786468
  • 21. Clark NP. Low-molecular-weight heparin use in the obese, elderly, and in renal insufficiency. Thromb Res. 2008;123(Suppl 1):58-61. PMID: 18809206
  • 22. Parikh S, Jakeman B, Walsh E, Townsend K, Burnett A. Adjusted-Dose Enoxaparin for VTE Preven-tion in the Morbidly Obese. J Pharm Tech. 2015;31:282–8. doi: 10.1177/8755122515593381
  • 23. Hoffman S, Braunreiter C. Reduced dosing of enoxaparin for venous thromboembolism in overweight and obese adolescents: a single institution retrospective review. Res Pract Thromb Haemost. 2017;1(2):188–93. PMID: 30046689
  • 24. Theerakittayakorn K, Bunprasert T. Differentiation Capacity of Mouse L929 Fibroblastic Cell Line Compare With Human Dermal Fibroblast. World Academy of Science, Engineering and Technolo-gy.2011;5:51-4. doi: 10.5281/zenodo.1084175
  • 25. Fareed J, Hoppensteadt D, Walenga J, Iqbal O, Ma Q, Jeske W, et al. Pharmacodynamic and phar-macokinetic properties of enoxaparin : implications for clinical practice. Clin Pharmacokinet. 2003;42:1043-57. PMID: 12959635.

Enoksaparin sodyum subkutan fibroblast için tam olarak güvenli midir?: Bir hücre kültürü çalışması

Year 2021, Volume 5, Issue 1, 18 - 23, 23.04.2021
https://doi.org/10.30565/medalanya.822679

Abstract

Amaç: Fibroblastlar, subkutan dokuda nispeten düşük miktarlara rağmen, hücreler arası bağlantıların sürekliliğinde, doku bütünlüğünün korunmasında ve dengeli bir fasiyal ağ oluşturmada kritik bir rol oynarlar. Enoksaparin sodyum, derin ven trombozunun profilaksisinde ve tedavisinde yaygın olarak kullanılmaktadır. Bu çalışmada, enoksaparin sodyumun fibroblast hücreleri üzerindeki sitotoksik ve apoptotik etkilerini in vitro bir modelde incelemeyi amaçladık.

Materyal ve metod: Bir fare modelinde, L929 fibroblast hücreleri enoksaparin sodyum 4000 IU, 2000 IU, 1000 IU, 500 IU ve 250 IU ile işleme tabi tutuldu. 48 saatte hücre morfolojisi değerlendirildi; hücre canlılığı metiltiyazol tetrazolyum deneyi ile analiz edildi ve apoptoz propidyum iyodür / akridin turuncu boyama ile değerlendirildi.

Bulgular: Test sonuçları, yüksek dozların (4000 IU, 2000 IU) sitotoksik etkiler yarattığını ve apoptotik morfolojiyi uyardığını gösterdi. Kontrol grubu ile karşılaştırıldığında, Dilüsyon III, IV ve V'de hücre canlılığında istatistiksel açıdan anlamlı bir fark bulunmadı.

Sonuç: Sonuçlarımıza göre, in vitro ortamda profilaktik doza rağmen, yüksek doz enoksaparin sitotoksik ve apoptotik etkiler göstermiştir. Uzun süreli yüksek doz enoksaparin sodyum, deri altı fibroblastların sayısını etkileyerek deri bütünlüğünü ve deri altı doku iyileşmesini bozabilir.

References

  • 1. Canales JF, Ferguson JJ. Low-Molecular-Weight Heparins Mechanisms, Trials,and Role in Contempo-rary Interventional Medicine. Am J Cardiovasc Drugs. 2008;8:15-25. PMID: 18303934
  • 2. Laurent P, Dussarat GV, Bonal J, Jego C, Talard P, Bouchiat C,et al. Low Molecular Weight Heparins A Guide to Their Optimum Use In Pregnancy. Drugs. 2002;62:463-77. PMID: 11827560
  • 3. Linkins LA. Heparin induced thrombocytopenia. BMJ. 2015;350:g7566. PMID: 25569604
  • 4. Atiq F, van den Bemt PM, Leebeek FW, van Gelder T, Versmissen J. A systematic review on the ac-cumulation of prophylactic dosages of low-molecular-weight heparins (LMWHs) in patients with renal insufficiency. Eur J Clin Pharmacol. 2015;71(8):921-9. PMID: 26071276
  • 5. An I, Harman M, Ibiloglu I, Bullous hemorrhagic dermatosis induced by enoxaparin. Indian Dermatol Online J. 2017;8(5):347-9. PMID: 28979868
  • 6. Bircher AJ, Harr T, Hohenstein L, Tsakiris DA. Hypersensitivity reactions to anticoagulant drugs: diag-nosis and management options. Allergy. 2006;61(12):1432-40. PMID: 17073874
  • 7. Ronceray S, Dinulescua M, Le Gall F, Polardc E, Dupuya A, Adamskia H. Enoxaparin- Induced DRESS Syndrome. Case Rep Dermatol. 2012;4(3):233–7. PMID:23185158
  • 8. Langevin HM, Cornbrooks CJ, Taatjes DJ. Fibroblasts form a body-wide cellular network. Histochem Cell Biol. 2004;122(1):7-15. PMID: 15221410
  • 9. Langevin HM, Bouffard NA, Churchill DL, Badger GJ. Connective tissue fibroblast response to acu-puncture: dose-dependent effect of bidirectional needle rotation. J Altern Complement Med. 2007;13(3):355-60. PMID: 17480137
  • 10. Fernandes IR, Russo FB, Pignatari GC, Evangelinellis MM, Tavolari S, Muotri AR ,et al. Fibroblast sources: Where can we get them?. Cytotechnology. 2016;68(2):223-8. PMID: 25060709
  • 11. Available at: https://www.anatomyatlases.org/MicroscopicAnatomy/Section03/ Plate0327.shtml Access date: 17.12.2019
  • 12. Gürpinar ÖA, Kubat E, Onur MA. Cytotoxic Activity of Apixaban on HeLa Cells: An in vitro Study. Hacettepe J Biol Chem. 2018;46(3):395–402. doi: 10.15671/HJBC.2018.247
  • 13. Kubat E, Gurpinar, Karasoy D, Onur MA. A link between cytotoxicity in cell culture and gastrointestinal side effects of oral anticoagulants: bench-to-bedside. Bratisl Med J. 2018;119(11):706-712. PMID: 30686004
  • 14. Hussain H, Raj LS., Ahmad S, Razak MFA, Mohamud WNW, Bakar J, et al. Determination of cell via-bility using acridine orange/propidium iodide dual-spectrofluorometry assay. Cogent Food & Agricul-ture. 2019:5:1582398:1-9. doi:10.1080/23311932.2019.1582398
  • 15. Ziani L, Chouaib S, Thiery J. Alteration of the Antitumor Immune Response by Cancer-Associated Fibroblasts. Front Immunol. 2018;9:414:1-14. PMID: 29545811
  • 16. Li B, Wang JH. Fibroblasts and myofibroblasts in wound healing: force generation and measurement. J Tissue Viability. 2011;20(4):108-20. PMID: 19995679
  • 17. Rippa AL, Kalabusheva EP, Vorotelyak EA. Regeneration of Dermis: Scarring and Cells Involved. Cells. 2019;8(6):607:1-30. PMID: 31216669
  • 18. Kobbi Z, Kraiem H, Benlasfar Z, Marouani A, Massoud T, Boubaker S, et al. Comparative subcutane-ous repeated toxicity study of enoxaparin products in rats. Regul Toxicol Pharmacol. 2017;84:9-17. PMID: 27965129
  • 19. Pföhler C, Müller CS, Pindur G, Eichler H, Schäfers HJ, Grundmann U, et al. Delayed-Type Heparin Allergy: Diagnostic Procedures and Treatment Alternatives- A Case Series Including 15 Patients. WAO Journal. 2008;1(12):194-9. PMID: 23282847
  • 20. Schindewolf M, Schwaner S, Wolter M, Kroll H, Recke A, Kaufmann R, et al. Incidence and causes of heparin-induced skin lesions. CMAJ. 2009;181(8):477-81. PMID: 19786468
  • 21. Clark NP. Low-molecular-weight heparin use in the obese, elderly, and in renal insufficiency. Thromb Res. 2008;123(Suppl 1):58-61. PMID: 18809206
  • 22. Parikh S, Jakeman B, Walsh E, Townsend K, Burnett A. Adjusted-Dose Enoxaparin for VTE Preven-tion in the Morbidly Obese. J Pharm Tech. 2015;31:282–8. doi: 10.1177/8755122515593381
  • 23. Hoffman S, Braunreiter C. Reduced dosing of enoxaparin for venous thromboembolism in overweight and obese adolescents: a single institution retrospective review. Res Pract Thromb Haemost. 2017;1(2):188–93. PMID: 30046689
  • 24. Theerakittayakorn K, Bunprasert T. Differentiation Capacity of Mouse L929 Fibroblastic Cell Line Compare With Human Dermal Fibroblast. World Academy of Science, Engineering and Technolo-gy.2011;5:51-4. doi: 10.5281/zenodo.1084175
  • 25. Fareed J, Hoppensteadt D, Walenga J, Iqbal O, Ma Q, Jeske W, et al. Pharmacodynamic and phar-macokinetic properties of enoxaparin : implications for clinical practice. Clin Pharmacokinet. 2003;42:1043-57. PMID: 12959635.

Details

Primary Language English
Subjects Surgery
Published Date 2021;5(1)
Journal Section Research Article
Authors

Emre KUBAT (Primary Author)
ANKARA T.C. SB. SBÜ. GÜLHANE EĞİTİM VE ARAŞTIRMA HASTANESİ
0000-0002-9884-8565
Türkiye


Aylin GÜRPINAR
HACETTEPE ÜNİVERSİTESİ, FEN FAKÜLTESİ, BİYOLOJİ BÖLÜMÜ
0000-0001-7055-1607
Türkiye


Gökşen ERTUĞRUL This is me
KARABÜK T.C. SB. KARABÜK ÜNİVERSİTESİ EĞİTİM VE ARAŞTIRMA HASTANESİ
0000-0002-5167-4780
Türkiye


Hakan IŞIK This is me
ANKARA T.C. SB. SBÜ. GÜLHANE EĞİTİM VE ARAŞTIRMA HASTANESİ
0000-0002-7602-4434
Türkiye


Duru KARASOY
HACETTEPE ÜNİVERSİTESİ, FEN FAKÜLTESİ, İSTATİSTİK BÖLÜMÜ
0000-0002-2258-4479
Türkiye


Mehmet Ali ONUR
HACETTEPE ÜNİVERSİTESİ, FEN FAKÜLTESİ, BİYOLOJİ BÖLÜMÜ
0000-0002-3630-7982
Türkiye

Publication Date April 23, 2021
Application Date November 6, 2020
Acceptance Date November 21, 2020
Published in Issue Year 2021, Volume 5, Issue 1

Cite

Vancouver Kubat E. , Gürpınar A. , Ertuğrul G. , Işık H. , Karasoy D. , Onur M. A. Is Enoxaparin Sodium Exactly Safe For Subcutaneous Fibroblast?: A Cell Culture Study. Acta Medica Alanya. 2021; 5(1): 18-23.

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