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The effects of strontium chloride on viability of mouse connective tissue fibroblast cells

Year 2013, Volume: 35 Issue: 1, 33 - 38, 22.01.2013

Abstract

Abstract

Aim. Strontium salts are effective and selective anti-irritants for chemically induced sensory irritation associated with stinging, burning, or itching. The aim of the present study was to determine the cytotoxic and/or proliferative effects of strontium chloride on fibroblast cell culture. Method. A mouse connective tissue fibroblast cell line, L929 (ATCC cell line, NCTC clone 929) was cultured. Fibroblast cell lines were examined with 20%, 10%, 5%, 2.5%, 1.25%, 0.6%, and 0.3% (w/v) concentrations of Strontium chloride hexahydrate (SrCl2.6H2O). The proliferation assay analyzed the number of viable cells by the cleavage of tetrazolium salts added to the culture medium, using the XTT labeling reagent. The optical density (OD) of the samples was compared with that of the control to obtain the percentage viability, as follows: cell viability (%)=[(OD450 (sample)/OD450 negative control))×100]. Results. The cytotoxicity value of strontium chloride for all concentrations (w/v) was compared with that of the control, and cytotoxicity levels were not higher than those of the controls. The level of viable cell was higher at 1.25% (w/v) than 2.5% (w/v) (p<0.05). Conclusion. Strontium chloride hexahydrate (SrCl2.6H2O) had no negative effect on cell viability at all the concentrations.

Keywords: Strontium chloride, fibroblast, cell culture, proliferation, cytotoxicity

 

Özet

Amaç. Stronsiyum tuzları yanma, batma ya da kaşıntıyla birlikteki kimyasal olarak uyarılmış sensoryal irritasyon için etkili ve seçici antiirritanlardır. Bu çalışmanın amacı fibroblast hücre kültürü üzerine stronsiyum kloridin sitotoksik ve/veya proliferatif etkilerini belirlemekti. Yöntem. Fare konnektif doku fibroblast hücrelerinin kültürü (L929 (ATCC cell line, NCTC clone 929) ) yapıldı. Fibroblast kültürü %20, %10, %5, %2,5 , %1,25 ve %0,6 ve %0,3 (w/v) konsantrasyonlarda stronsiyum klorid heksahidrat ile muamele edildi. Proliferasyon düzeneğinde XTT işaretli reagen kullanılarak kültür ortamına eklenen tetrazolyum tuzlarının klivajı yoluyla canlı hücre sayısı analiz edildi. Örneklerin optik dansitesi kontrol grubuyla karşılaştırıldı ve şu formül kullanıldı: Hücre canlılığı (%)=[(OD450 (örnek)/OD450 (negatif kontrol))×100]. Bulgular. Stronsiyum klorid sitotoksisite ve hücre canlılığı açısından kontrol grubuyla karşılaştırıldığında istatistiksel olarak anlamlı fark bulundu (p<0.05). %1.25 konsantrasyonda canlı hücre sayısı diğer konsantrasyonlardan daha fazlaydı (p<0.05). Tüm konsantrasyonda sitotoksisite değeri kontrol grubundan daha fazla değildi. Sonuç. Stronsiyum klorid heksahidratın hiçbir konsantrasyonda hücre canlılığı üzerine olumsuz etkisi yoktu.

Anahtar sözcükler: Stronsiyum klorid, fibroblast, hücre kültürü, proliferasyon, sitotoksisite

References

  • Burlet N, Reginster JY. Strontium ranelate: The first dual acting treatment for postmenopausal osteoporosis. Clin Orthop Relat Res 2006; 443; 55-60.
  • Ammann P. Strontium ranelate: a physiological approach for an improved bone quality. Bone 2006; 38: 15-8.
  • Price TD, Swick RW, Chase EP. Bone chemistry and prehistoric diet: strontium studies of laboratory rats. Am J Phsic Anthropol 1986; 70: 365-75.
  • Er K, Polat ZA, Ozan F, Taşdemir T, Sezer U, Siso SH. Cytotoxicity analysis of strontium ranelate on cultured human periodontal ligament fibroblasts: a preliminary report. J Formos Med Assoc 2008; 107: 609-15.
  • Hahn GS. Strontium is a potent and selective inhibitor of sensory irritation. Dermatol Surg 1999; 25: 689-94.
  • Zhai H, Hannon W, Hahn GS, Pelosi A, Harper RA, Maibach HI. Strontium nitrate suppresses chemically-induced sensory irritation in humans. Contact Dermatitis 2000; 42: 98-100.
  • Gu C, Cooper DM. Ca(2+), Sr(2+), and Ba(2+) identify distinct regulatory sites on adenylyl cyclase (AC) types VI and VIII and consolidate the apposition of capacitative cation entry channels and Ca(2+)-sensitive ACs. J Biol Chem 2000; 275: 6980-6.
  • Concise International Chemical assessment document 77. Strontium and strontium compounds. First draft prepared by Mr Peter Watts, Toxicology Advice & Consulting Ltd, Sutton, England; and Mr Paul Howe, Centre for Ecology and Hydrology, Monks Wood, England. World Health Organization. 2010.
  • Toxicological profile for strontium. Atlanta, GA, United States Department of Health and Human Services, Public Health Service, Agency for Toxic Substances and Disease Registry (http://www.atsdr.cdc.gov/Toxprofiles/TP.asp?id=656&tid=120). ATSDR 2004 (Accesed on February 21. 2013).
  • Amado A, Taylor JS, Sood A. Irritant contact dermatitis. In: Wolff K, Goldsmith LA, Katz SI, Gilchrest BA, Paller ES, Leffell DJ (eds), Fitzpatrick’s Dermatology in general Medicine, 7th edition, New York, McGraw Hill Medical. 2008; 395-401.
  • Jacobs JJ, Lehé CL, Hasegawa H, Elliott GR, Das PK. Skin irritants and contact sensitizers induce Langerhans cell migration and maturation at irritant concentration. Exp Dermatol 2006; 15: 432-40.
  • Celerier P, Richard A, Litoux P, Dreno B. Modulatory effects of selenium and strontium salts on keratinocyte-derived inflammatory cytokines. Arch Dermatol Res 1995; 287: 680-2.
  • Caverzasio J, Thouverey C. Activation of FGF receptors is a new mechanism by which strontium ranelate induces osteoblastic cell growth. Cell Physiol Biochem 2011; 27: 243-50.
  • Lymperi S, Horwood N, Marley S, Gordon MY, Cope AP, Dazzi F. Strontium can increase some osteoblasts without increasing hematopoietic stem cells. Blood 2008; 111: 1173-81.
  • Ousey K, Mclntosh C. Physiology of wound healing. Ousey K, Mclntosh C (eds), Hoboken, Wiley Ltd. 2008; 25-46.
  • Moens W, Vokaer A, Kram R. Cyclic AMP and cyclic GMP concentrations in serum- and density-restricted fibroblast cultures. Prot Nat Acad Sci 1975; 72: 1063Fisch C, Attia M, Dargent F, de Jouffrey S, Dupin-Roger I, Claude JR. Preclinical assessment of gastrooesophageal tolerance of the new antiosteoporotic drug strontium ranelate: an endoscopic study in monkeys. Basic Clin Pharmacol Toxicol 2006; 98: 442-6.
  • Kroes R, den Tonkelaar EM, Minderhoud A, Speijers GJ, Vonk-Visser DM, Berkvens JM, van Esch GJ. Short-term toxicity of strontium chloride in rats. Toxicology 1977; 7: 11-21.
  • Skoryna SC. Effects of oral supplementation with stable strontium. Canadian Med Assoc J 1981; 125: 703-12.

Original research-Orijinal araştırma

Year 2013, Volume: 35 Issue: 1, 33 - 38, 22.01.2013

Abstract

Amaç. Stronsiyum tuzları yanma, batma ya da kaşıntıyla birlikteki kimyasal olarak uyarılmış sensoryal irritasyon için etkili ve seçici antiirritanlardır. Bu çalışmanın amacı fibroblast hücre kültürü üzerine stronsiyum kloridin sitotoksik ve/veya proliferatif etkilerini belirlemekti. Yöntem. Fare konnektif doku fibroblast hücrelerinin kültürü (L929 (ATCC cell line, NCTC clone 929) ) yapıldı. Fibroblast kültürü %20, %10, %5, %2,5 , %1,25 ve %0,6 ve %0,3 (w/v) konsantrasyonlarda stronsiyum klorid heksahidrat ile muamele edildi. Proliferasyon düzeneğinde XTT işaretli reagen kullanılarak kültür ortamına eklenen tetrazolyum tuzlarının klivajı yoluyla canlı hücre sayısı analiz edildi. Örneklerin optik dansitesi kontrol grubuyla karşılaştırıldı ve şu formül kullanıldı: Hücre canlılığı (%)=[(OD450 (örnek)/OD450 (negatif kontrol))×100]. Bulgular. Stronsiyum klorid heksahidrat sitotoksisite açısından kontrol grubuna göre farklılık göstermemiştir (p>0.05). %1.25 konsantrasyonda canlı hücre sayısı diğer konsantrasyonlardan daha fazlaydı (p<0.05). Tüm konsantrasyonda sitotoksisite değeri kontrol grubundan daha fazla değildi. Sonuç. Stronsiyum klorid heksahidratın fibroblast hücre kültüründe, hiçbir konsantrasyonda hücre canlılığı üzerine olumsuz etkisi saptanmamıştır.

References

  • Burlet N, Reginster JY. Strontium ranelate: The first dual acting treatment for postmenopausal osteoporosis. Clin Orthop Relat Res 2006; 443; 55-60.
  • Ammann P. Strontium ranelate: a physiological approach for an improved bone quality. Bone 2006; 38: 15-8.
  • Price TD, Swick RW, Chase EP. Bone chemistry and prehistoric diet: strontium studies of laboratory rats. Am J Phsic Anthropol 1986; 70: 365-75.
  • Er K, Polat ZA, Ozan F, Taşdemir T, Sezer U, Siso SH. Cytotoxicity analysis of strontium ranelate on cultured human periodontal ligament fibroblasts: a preliminary report. J Formos Med Assoc 2008; 107: 609-15.
  • Hahn GS. Strontium is a potent and selective inhibitor of sensory irritation. Dermatol Surg 1999; 25: 689-94.
  • Zhai H, Hannon W, Hahn GS, Pelosi A, Harper RA, Maibach HI. Strontium nitrate suppresses chemically-induced sensory irritation in humans. Contact Dermatitis 2000; 42: 98-100.
  • Gu C, Cooper DM. Ca(2+), Sr(2+), and Ba(2+) identify distinct regulatory sites on adenylyl cyclase (AC) types VI and VIII and consolidate the apposition of capacitative cation entry channels and Ca(2+)-sensitive ACs. J Biol Chem 2000; 275: 6980-6.
  • Concise International Chemical assessment document 77. Strontium and strontium compounds. First draft prepared by Mr Peter Watts, Toxicology Advice & Consulting Ltd, Sutton, England; and Mr Paul Howe, Centre for Ecology and Hydrology, Monks Wood, England. World Health Organization. 2010.
  • Toxicological profile for strontium. Atlanta, GA, United States Department of Health and Human Services, Public Health Service, Agency for Toxic Substances and Disease Registry (http://www.atsdr.cdc.gov/Toxprofiles/TP.asp?id=656&tid=120). ATSDR 2004 (Accesed on February 21. 2013).
  • Amado A, Taylor JS, Sood A. Irritant contact dermatitis. In: Wolff K, Goldsmith LA, Katz SI, Gilchrest BA, Paller ES, Leffell DJ (eds), Fitzpatrick’s Dermatology in general Medicine, 7th edition, New York, McGraw Hill Medical. 2008; 395-401.
  • Jacobs JJ, Lehé CL, Hasegawa H, Elliott GR, Das PK. Skin irritants and contact sensitizers induce Langerhans cell migration and maturation at irritant concentration. Exp Dermatol 2006; 15: 432-40.
  • Celerier P, Richard A, Litoux P, Dreno B. Modulatory effects of selenium and strontium salts on keratinocyte-derived inflammatory cytokines. Arch Dermatol Res 1995; 287: 680-2.
  • Caverzasio J, Thouverey C. Activation of FGF receptors is a new mechanism by which strontium ranelate induces osteoblastic cell growth. Cell Physiol Biochem 2011; 27: 243-50.
  • Lymperi S, Horwood N, Marley S, Gordon MY, Cope AP, Dazzi F. Strontium can increase some osteoblasts without increasing hematopoietic stem cells. Blood 2008; 111: 1173-81.
  • Ousey K, Mclntosh C. Physiology of wound healing. Ousey K, Mclntosh C (eds), Hoboken, Wiley Ltd. 2008; 25-46.
  • Moens W, Vokaer A, Kram R. Cyclic AMP and cyclic GMP concentrations in serum- and density-restricted fibroblast cultures. Prot Nat Acad Sci 1975; 72: 1063Fisch C, Attia M, Dargent F, de Jouffrey S, Dupin-Roger I, Claude JR. Preclinical assessment of gastrooesophageal tolerance of the new antiosteoporotic drug strontium ranelate: an endoscopic study in monkeys. Basic Clin Pharmacol Toxicol 2006; 98: 442-6.
  • Kroes R, den Tonkelaar EM, Minderhoud A, Speijers GJ, Vonk-Visser DM, Berkvens JM, van Esch GJ. Short-term toxicity of strontium chloride in rats. Toxicology 1977; 7: 11-21.
  • Skoryna SC. Effects of oral supplementation with stable strontium. Canadian Med Assoc J 1981; 125: 703-12.
There are 18 citations in total.

Details

Primary Language English
Journal Section Basic Science Research Articles
Authors

Melih Akyol

Zübeyde Akın Polat

Sedat Özçelik

Özge Kaya

Publication Date January 22, 2013
Published in Issue Year 2013Volume: 35 Issue: 1

Cite

AMA Akyol M, Akın Polat Z, Özçelik S, Kaya Ö. The effects of strontium chloride on viability of mouse connective tissue fibroblast cells. CMJ. March 2013;35(1):33-38.