Clinical, histopathological and immunohistochemical evaluation of the effects of topical NPH-insulin on full-thickness open wounds: An in vivo study in diabetic and non-diabetic mice
Abstract
In diabetic patients, wound healing is disrupted due to impaired local circulation and infection or healing becomes impossible due to the gangrene. Recently, wound treatment models based on the use of topical insulin have been the subject of some studies. In this study, we aimed to evaluate the efficacy of topical NPH insulin use on wound healing for the treatment of full-thickness open wounds in diabetic and non-diabetic mice. A total of 72, non-diabetic and diabetic adult male Swiss Albino mice were used in the study. A full-thickness skin excision with a diameter of 1 cm was practiced on the back of all mice. Animals in diabetic and nondiabetic groups were divided into two sub-groups as NPH insulin used and control group. After the wound surface areas were digitally measured, the animals were sacrificed for histopathological and immunohistochemical examinations. Wound measurements revealed a significant reduction in the non-diabetic insulin treatment group at 7th and 14th days and at 3rd, 7th and 14th days of treatment in the diabetic insulin treatment group. Histopathological examinations showed that there was a significant increase in granulomatous inflammation in the wound site in the non-diabetic insulin treatment group at 3rd days and in the diabetic insulin treatment group at 14thdays. Immunohistochemical analyses showed that there was an increase in α-SMA at 3rd days and a decrease in TGF-β1 at 14th days in the diabetic insulin treatment group, while there was a significant decrease in IGF-I at 7th days in all diabetic groups. Topical NPH insulin application significantly accelerated healing in the treatment of diabetic and non-diabetic wounds with tissue loss in mice
Keywords
References
- Akasaka Y, Ono I, Tominaga A, et al. (2007): Basic fibroblast growth factor in an artificial dermis promotes apoptosis and inhibits expression of alpha-smooth muscle actin, leading to reduction of wound contraction. Wound Repair Regen, 15, 378-389.
- Akbarzadeh A, Norouzian D, Mehrabi MR, et al. (2007): Induction of diabetes by streptozotocin in rats. Indian J Clin Biochem, 22, 60-64.
- Apikoglu-Rabus S, Izzettin FV, Turan P, et al. (2010): Effect of topical insulin on cutaneous wound healing in rats with or without acute diabetes. Clin Exp Dermatol, 35, 180- 185.
- Armstrong DG, Lavery LA (2005): Negative pressure wound theraphy after partial diabetic foot amputation: a multicentre, randomised controlled trial. Lancet, 366, 1704-10.
- Aydin F, Kaya A, Karapinar L, et al. (2013): IGF-1 increases with hyperbaric oxygen theraphy and promotes wound healing in diabetic foot ulcers. J Diabetes Res, doi: 10.1155/2013/567834.
- Bairy KL, Abhinav R, Satyam SM (2014): Evaluation of burn wound healing activity of topical regular insulin in non-diabetic and streptozocin-induced diabetic rats. Int J Pharm Sci, 6, 127-130.
- Belfield WO, Golinsky S, Compton MD (1970): The use of insulin in open-wound healing. Vet Med Small Anim Clin, 65, 455-460.
- Bennett NT, Schultz GS (1993): Growth factors and wound healing: Part II. Role in normal and chronic wound healing. Am J Surg, 166, 74-81.
- Carrico TJ, Mehrhof AI Jr, Cohen IK (1984): Biology of wound healing. Surg Clin North Am, 64, 721-733.
- Darby IA, Bisucci T, Hewitson TD, et al. (1997): Apoptosis is increased in a model of diabetes-impaired wound healing in genetically diabetic mice. Int J Biochem Cell Biol, 29, 191-200.
- Gabbiani G (2004): The evolution of the myofibroblast concept: A key cell for wound healing and fibrotic diseases. J Gerontol, 52, 280-282.
- Gal P, Kilik R, Mokry B, et al. (2008): Simple method of open skin wound healing model in corticosteroid-treated and diabetic rats: Standardization of semi-quantitative and quantitative histological assessments. Vet Med Praha, 53, 652-659.
- Galiano RD, Tepper OM, Pelo CR, et al. (2004): Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells. Am J Pathol, 164, 1935-1947.
- Goren I, Müller E, Schiefelbein D, et al. (2009): Akt1 controls insulin-driven VEGF biosynthesis from keratinocytes: implications for normal and diabetes-impaired skin repair in mice. J Invest Dermatol, 129, 752-764.
- Greenway SE, Filler LE, Greenway FL (1999): Topical insulin in wound healing: a randomised, double-blind placebo-controlled trial. J Wound Care, 8, 526-528.
- Grotendorst GR, Rahmanie H, Duncan MR (2004): Combinatorial signaling pathways determine fibroblast proliferation and myofibroblast differentiation. FASEB J, 18, 469-479.
- Gül Satar NY, Cangül İT, Topal A, et al. (2014): Effects of Ankaferd Blood Stopper (ABS) and topical tripeptide copper complex (TCC) on wound healing in rats: An experimental study. Kafkas Univ Vet Fak Derg, 20, 545-551.
- Hinz B (2007): Formation and function of the myofibroblast during tissue repair. J Invest Dermatol, 127, 526-537.
- Hinz B, Celetta G, Tomasek JJ, et al. (2001): Alpha- smooth muscle actin expression upregulates fibroblast contractile activity. Mol Biol Cell, 12, 2730-2741.
- Hunt TK (1990): Basic principles of wound healing. J Trauma, 30, 122-128.
- Kargın S, Taştekin D, Kılıç K, et al. (2015): The effects of topical insulin application on wound healing. Eur J Gen Med, 12, 302-306.
- Koç B, Özaydın İ, Özba B, et al. (1990): Clinical observations on the application of local insulin and pinch graft in gangrenous-open wound. 2nd National Veterinary Surgery Congress, 60-65, October 1-2, Alata, Mersin, Turkey.
- Lima MHM, Caricilli AM, Abreu LL, et al. (2012): Topical insulin accelerates wound healing in diabetes by enhancing the AKT and ERK pathways: A double-blind placebo-controlled clinical trial. PLoS One, 7, doi: 10.1371/journal.pone.0036974.
- Liu Y, Petreaca M, Yao M, et al. (2009): Cell and molecular mechanisms of keratinocyte function stimulated by insulin during wound healing. BMC Cell Biol, 10, doi: 10.1186/1471-2121-10-1.
- Madibally SV, Solomon V, Mitchell RN, et al. (2003): Influence of insulin therapy on burn wound healing in rats. J Surg Res, 109, 92-100.
- Moulin V, Auger FA, Garrel D, et al. (2000): Role of wound healing myofibroblasts on re-epithelization of human skin. Burns, 26, 3-12.
- Mutsaers SE, Bishop JE, McGrouther G, et al. (1997): Mechanisms of tissue repair: From wound healing to fibrosis. Int J Biochem Cell Biol, 29, 5-17.
- Ng PC, Hinz B, Swartz MA (2005): Interstitial fluid flow induces myofibroblast differentiation and collagen alignment in vitro. J Cell Sci, 118, 4731-39.
- Oruç E, Kara A, Can İ, et al. (2012): Caspase-3 and CD68 Immunoreactivity in lymphoid tissues and haematology of rats exposed to cisplatin and L-carnitine. Kafkas Univ Vet Fak Derg, 18, 871-878.
- Ömeroğlu S, Çam M, Erdoğan D, et al. (2003): Immunohistochemical demonstration of insulin like growth factor-I localisation in the skin wounds of streptozotocin administered experimental diabetic rats. Duzce Med J, 5, 5-8.
- Özaydın İ (2004): Traumatic Wounds. 128-137. In: Özaydın İ (Ed), Veterinary Emergency Clinic: First Aid, Transporting, Medical Intervention. Eser Ofset, Erzurum.
- Rezvani O, Shabbak E, Aslani A, et al. (2009): A randomized, double-blind, placebo controlled trial to determine the effects of topical insulin on wound healing. Ostomy Wound Manage, 55, 22-28.
- Stadelmann WK, Digenis AG, Tobin GR (1998): Physiology and healing dynamics of chronic cutaneous wounds. Am J Surg, 176, 26-38.
- Taçyıldız HI, Mızraklı S, Aban N, et al. (1998): The effect of collagenase in wound healing. Turkish J Trauma Emerg Surg, 4, 7-11.
- Thorey IS, Hinz B, Hoeflich A, et al. (2004): Transgenic mice reveal novel activities of growth hormone in wound repair, angiogenesis and myofibroblast differentiation. J Biol Chem, 279, 26674-26684.
- Wu Y, Chen L, Scott PG, et al. (2007): Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells, 25, 2648-2659.
- Yoruk O, Dane S, Ucuncu H, et al. (2009): Stereological evaluation tomography via the Cavalieri method: Correlation between tumor volume and number of neck lymph node metastases. J Craniofac Surg, 20, 1504-07. using computed Geliş tarihi: 29.01.2017 / Kabul tarihi: 04.06.2017 Address for correspondence: Başak KURT Kafkas University, Faculty of Veterinary Medicine, Department of Surgery, TR 36100, Kars, Turkey. Phone: 904742426836 e-mail:basakyakin@hotmail.com
Tam kalınlıklı açık yaralarda topikal NPH insülinin etkilerinin klinik, histopatolojik veimmunohistokimyasal olarak değerlendirilmesi: Diyabetik ve non-diyabetik farelerde in vivo birçalışma
Abstract
Diyabetik hastalarda lokal sirkülasyonun bozulması ve enfeksiyon nedeniyle yara iyileşmesi sekteye uğrar veya gangren şekillenerek iyileşme imkansız bir tabloya dönüşür. Son zamanlarda topikal insülinin kullanımına dayalı yara sağaltım modelleri araştırmalara konu olmaktadır. Sunulan çalışma ile diyabetik ve non-diyabetik farelerde oluşturulan tam kalınlıkta açık yara sağaltımı amacıyla topikal NPH insülin kullanımının, yara iyileşmesi üzerine etkinliğinin değerlendirilmesi amaçlandı. Çalışmada, non-diyabetik ve diyabetik 72 adet erişkin erkek Swiss Albino fare kullanıldı. Tüm farelerde, genel anestezi eşliğinde sırt bölgesinde 1 cm çapında tam kalınlıkta deri eksizyonu ile yara oluşturuldu. Diyabetik ve non-diyabetik gruptaki hayvanlar NPH insulin ve kontrol grubu olmak üzere ikişer alt gruba ayrıldı. Üç, 7 ve 14. günlerde dijital ortamda yara yüzey alanı ölçümleri yapıldıktan sonra, histopatolojik ve immunohistokimyasal incelemeler için hayvanlar sakrifiye edildi. Yara alanı ölçümlerinde, non-diyabetik insülin grubunda tedavinin 7 ve 14. günlerinde, diyabetik insülin grubunda ise 3, 7 ve 14. günlerde kontrol gruplarına göre istatistiksel olarak anlamlı bir küçülme olduğu gözlendi. Histopatolojik incelemeler, granülamatöz yapıların non-diyabetik insülin grubunun yara bölgesinde kendi kontrol grubuna göre 3. günde, diyabetik grubun kendi kontrol grubuna göre ise 14. günde belirgin artış gösterdiğini ortaya koydu. İmmunohistokimyasal analizlerde diyabetik insülin grubunda 3. gün α-SMA’da artış, 14. gün TGF-β1’de düşüş dikkati çekerken, tüm diyabetik gruplarda 7. gün IGF-I’in önemli oranda düşüş gösterdiği belirlendi. Sunulan çalışma ile diyabetik ve non-diyabetik doku kayıplı yaraların sağaltımında topikal insülin uygulamasının iyileşmeyi belirgin şekilde hızlandırdığı ortaya konuldu
Keywords
References
- Akasaka Y, Ono I, Tominaga A, et al. (2007): Basic fibroblast growth factor in an artificial dermis promotes apoptosis and inhibits expression of alpha-smooth muscle actin, leading to reduction of wound contraction. Wound Repair Regen, 15, 378-389.
- Akbarzadeh A, Norouzian D, Mehrabi MR, et al. (2007): Induction of diabetes by streptozotocin in rats. Indian J Clin Biochem, 22, 60-64.
- Apikoglu-Rabus S, Izzettin FV, Turan P, et al. (2010): Effect of topical insulin on cutaneous wound healing in rats with or without acute diabetes. Clin Exp Dermatol, 35, 180- 185.
- Armstrong DG, Lavery LA (2005): Negative pressure wound theraphy after partial diabetic foot amputation: a multicentre, randomised controlled trial. Lancet, 366, 1704-10.
- Aydin F, Kaya A, Karapinar L, et al. (2013): IGF-1 increases with hyperbaric oxygen theraphy and promotes wound healing in diabetic foot ulcers. J Diabetes Res, doi: 10.1155/2013/567834.
- Bairy KL, Abhinav R, Satyam SM (2014): Evaluation of burn wound healing activity of topical regular insulin in non-diabetic and streptozocin-induced diabetic rats. Int J Pharm Sci, 6, 127-130.
- Belfield WO, Golinsky S, Compton MD (1970): The use of insulin in open-wound healing. Vet Med Small Anim Clin, 65, 455-460.
- Bennett NT, Schultz GS (1993): Growth factors and wound healing: Part II. Role in normal and chronic wound healing. Am J Surg, 166, 74-81.
- Carrico TJ, Mehrhof AI Jr, Cohen IK (1984): Biology of wound healing. Surg Clin North Am, 64, 721-733.
- Darby IA, Bisucci T, Hewitson TD, et al. (1997): Apoptosis is increased in a model of diabetes-impaired wound healing in genetically diabetic mice. Int J Biochem Cell Biol, 29, 191-200.
- Gabbiani G (2004): The evolution of the myofibroblast concept: A key cell for wound healing and fibrotic diseases. J Gerontol, 52, 280-282.
- Gal P, Kilik R, Mokry B, et al. (2008): Simple method of open skin wound healing model in corticosteroid-treated and diabetic rats: Standardization of semi-quantitative and quantitative histological assessments. Vet Med Praha, 53, 652-659.
- Galiano RD, Tepper OM, Pelo CR, et al. (2004): Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells. Am J Pathol, 164, 1935-1947.
- Goren I, Müller E, Schiefelbein D, et al. (2009): Akt1 controls insulin-driven VEGF biosynthesis from keratinocytes: implications for normal and diabetes-impaired skin repair in mice. J Invest Dermatol, 129, 752-764.
- Greenway SE, Filler LE, Greenway FL (1999): Topical insulin in wound healing: a randomised, double-blind placebo-controlled trial. J Wound Care, 8, 526-528.
- Grotendorst GR, Rahmanie H, Duncan MR (2004): Combinatorial signaling pathways determine fibroblast proliferation and myofibroblast differentiation. FASEB J, 18, 469-479.
- Gül Satar NY, Cangül İT, Topal A, et al. (2014): Effects of Ankaferd Blood Stopper (ABS) and topical tripeptide copper complex (TCC) on wound healing in rats: An experimental study. Kafkas Univ Vet Fak Derg, 20, 545-551.
- Hinz B (2007): Formation and function of the myofibroblast during tissue repair. J Invest Dermatol, 127, 526-537.
- Hinz B, Celetta G, Tomasek JJ, et al. (2001): Alpha- smooth muscle actin expression upregulates fibroblast contractile activity. Mol Biol Cell, 12, 2730-2741.
- Hunt TK (1990): Basic principles of wound healing. J Trauma, 30, 122-128.
- Kargın S, Taştekin D, Kılıç K, et al. (2015): The effects of topical insulin application on wound healing. Eur J Gen Med, 12, 302-306.
- Koç B, Özaydın İ, Özba B, et al. (1990): Clinical observations on the application of local insulin and pinch graft in gangrenous-open wound. 2nd National Veterinary Surgery Congress, 60-65, October 1-2, Alata, Mersin, Turkey.
- Lima MHM, Caricilli AM, Abreu LL, et al. (2012): Topical insulin accelerates wound healing in diabetes by enhancing the AKT and ERK pathways: A double-blind placebo-controlled clinical trial. PLoS One, 7, doi: 10.1371/journal.pone.0036974.
- Liu Y, Petreaca M, Yao M, et al. (2009): Cell and molecular mechanisms of keratinocyte function stimulated by insulin during wound healing. BMC Cell Biol, 10, doi: 10.1186/1471-2121-10-1.
- Madibally SV, Solomon V, Mitchell RN, et al. (2003): Influence of insulin therapy on burn wound healing in rats. J Surg Res, 109, 92-100.
- Moulin V, Auger FA, Garrel D, et al. (2000): Role of wound healing myofibroblasts on re-epithelization of human skin. Burns, 26, 3-12.
- Mutsaers SE, Bishop JE, McGrouther G, et al. (1997): Mechanisms of tissue repair: From wound healing to fibrosis. Int J Biochem Cell Biol, 29, 5-17.
- Ng PC, Hinz B, Swartz MA (2005): Interstitial fluid flow induces myofibroblast differentiation and collagen alignment in vitro. J Cell Sci, 118, 4731-39.
- Oruç E, Kara A, Can İ, et al. (2012): Caspase-3 and CD68 Immunoreactivity in lymphoid tissues and haematology of rats exposed to cisplatin and L-carnitine. Kafkas Univ Vet Fak Derg, 18, 871-878.
- Ömeroğlu S, Çam M, Erdoğan D, et al. (2003): Immunohistochemical demonstration of insulin like growth factor-I localisation in the skin wounds of streptozotocin administered experimental diabetic rats. Duzce Med J, 5, 5-8.
- Özaydın İ (2004): Traumatic Wounds. 128-137. In: Özaydın İ (Ed), Veterinary Emergency Clinic: First Aid, Transporting, Medical Intervention. Eser Ofset, Erzurum.
- Rezvani O, Shabbak E, Aslani A, et al. (2009): A randomized, double-blind, placebo controlled trial to determine the effects of topical insulin on wound healing. Ostomy Wound Manage, 55, 22-28.
- Stadelmann WK, Digenis AG, Tobin GR (1998): Physiology and healing dynamics of chronic cutaneous wounds. Am J Surg, 176, 26-38.
- Taçyıldız HI, Mızraklı S, Aban N, et al. (1998): The effect of collagenase in wound healing. Turkish J Trauma Emerg Surg, 4, 7-11.
- Thorey IS, Hinz B, Hoeflich A, et al. (2004): Transgenic mice reveal novel activities of growth hormone in wound repair, angiogenesis and myofibroblast differentiation. J Biol Chem, 279, 26674-26684.
- Wu Y, Chen L, Scott PG, et al. (2007): Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells, 25, 2648-2659.
- Yoruk O, Dane S, Ucuncu H, et al. (2009): Stereological evaluation tomography via the Cavalieri method: Correlation between tumor volume and number of neck lymph node metastases. J Craniofac Surg, 20, 1504-07. using computed Geliş tarihi: 29.01.2017 / Kabul tarihi: 04.06.2017 Address for correspondence: Başak KURT Kafkas University, Faculty of Veterinary Medicine, Department of Surgery, TR 36100, Kars, Turkey. Phone: 904742426836 e-mail:basakyakin@hotmail.com
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Other ID | JA48EU73KJ |
| Journal Section | Research Article |
| Authors | |
| Publication Date | September 1, 2018 |
| Published in Issue | Year 2018 |