{"id":29372,"date":"2019-12-28T11:30:33","date_gmt":"2019-12-28T11:30:33","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=29372"},"modified":"2020-04-22T05:33:23","modified_gmt":"2020-04-22T05:33:23","slug":"a-probe-into-prohealing-potential-of-glyceryl-trinitrate","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol12no4\/a-probe-into-prohealing-potential-of-glyceryl-trinitrate\/","title":{"rendered":"A Probe into Prohealing Potential of Glyceryl Trinitrate"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

Wound healing is a complex, delicate process which consists of series of cellular and biochemical events leading to re-establishment of anatomical continuity. Generally wounds heals without complications. There are many factors which affect wound healing. These factors may interfere with one of the phases of wound healing. There may be impaired wound healing, which leads to chronicity of wound eg. Chronic anal fissure, diabetic foot wound, venous ulcers. Chronic wound requires several days of treatment to achieve satisfactory healing without complications. The need for improving impaired wound healing has resulted in continued research on wound healing in human as well as animals.<\/p>\n

Anal fissure is a painful, chronic, non-healing wound. It\u2019s chronicity is due to spasm of anal sphincter and local ischemia.1,2 <\/sup>\u00a0Commonly, anal fissure is treated surgically by sphincterotomy to eliminate sphincter spasm.\u00a0 Glyceryl trinitrate(GTN) is a non-surgical alternative treatment due to its spasmolytic action.3,4<\/sup><\/p>\n

GTN is also used to promote wound healing.5,6.<\/sup> GTN has been used in chronic anal fissure and compared with calcium channel blockers for its therapeutic effect.7<\/sup>.<\/p>\n

The prohealing effect of \u00a0GTN\u00a0 is believed to be\u00a0 due to smooth muscle relaxation resulting in mitigation of sphincter spasm and local ischemia.8,9<\/sup> Since wound healing is a complex phenomenon, there is a possibility that wound healing potential of GTN may not be solely due to smooth muscle relaxant action. Therefore, the present study was undertaken to investigate, whether GTN has any wound healing property apart from its spasmolytic action. We have also studied the effect of GTN ointment base, to see if wound healing is\u00a0 due to the drug itself or is potentiated by the base.<\/p>\n

Aim and Objectives<\/strong><\/p>\n

To determine wound healing potential of GTN.<\/p>\n

Materials and methods<\/strong><\/p>\n

Study was carried out in 3 groups of rats ( 6 rats per group) for\u00a0 wound healing potential of Glyceryl trinitrate at Dept of Pharmacology and Central Animal House, Bharati Vidyapeeth Deemed University, Medical College & Hospital Sangli.<\/p>\n

Synopsis was discussed in IAEC (Institutional Animal Ethical Committee). IAEC approved the project, IAEC approval registration no.- BVDUMC&H, Sangli CAH\/ 2015\/11<\/p>\n

Total 18 male Wistar rats weighing 150-250 gm were taken. These were divided into three groups, 6 rats per group.<\/p>\n

Chemicals \/ Drugs:\u00a0 surgical material used<\/strong><\/p>\n

Glyceryl trinitrate ointment- 0.2% (GTN) \u2013Troikaa Pharmaceuticals Ltd.<\/p>\n

Ointment base (OB) – Troikaa Pharmaceuticals Ltd.<\/p>\n

Thiopental sodium, Spirit and Normal Saline, scissor, scalpel blade, artery forcep<\/p>\n

Table 1:<\/strong>\u00a0Group of animals and Drug Treatment Schedule for <\/strong>excisional wound model<\/strong><\/p>\n\n\n\n\n\n\n
Group No.<\/strong><\/td>\nNo. of\u00a0 Animals<\/strong><\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Drugs\u00a0\u00a0 <\/strong><\/td>\n<\/tr>\n
1<\/strong><\/td>\n6<\/td>\nControl (Normal Saline)<\/td>\n<\/tr>\n
2<\/strong><\/td>\n6<\/td>\n0.2% Glyceryl trinitrate ointment<\/td>\n<\/tr>\n
3<\/strong><\/td>\n6<\/td>\nGTN Ointment Base<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Methodology<\/strong><\/p>\n

It was a double blinded study. Randomization was done by simple random method.<\/p>\n

The animals bearing experimental wounds were treated with Normal Saline, GTN, GTN Ointment Base.<\/p>\n

Wounding Procedure<\/strong><\/p>\n

The nape of neck of rat was shaved. All the rats were starved overnight with water ad libitum <\/em>and the next day, surgical intervention under general anaesthesia was done to create excisional wound.<\/p>\n

About 500 mm2<\/sup> full thickness skin was excised, circular in shape with scalpel blade on the nape of the neck by using method of\u00a0 Morton and Malone10<\/sup>.<\/sub><\/p>\n

Drug Schedule<\/strong><\/p>\n

18 rats were wounded. 6 rats in each group received Normal Saline, GTN and GTN Ointment Base.\u00a0 Drug application was done locally once daily from the next day of wounding. The drug application was continued till the wound healed completely, as shown by total epithelization of the wound.<\/p>\n

Monitoring of healing<\/strong><\/p>\n

Excisional wounds<\/strong><\/p>\n

Two parameters viz; contraction of wound and epithelization period were monitored. The wound contraction was accomplished by periodical (every 4th <\/sup>day post wounding) recording of wound size by planimetry or by tracing the wound area on polythene paper first and subsequently on mm2 <\/sup>paper every 4th<\/sup> day. The degree of wound healing was calculated as percentage closure of the original wound area using the following formula-<\/p>\n

Percentage closure =A0 -Ad\/ A0 \u0445 100<\/p>\n

Where A0 = wound area on zero day and<\/p>\n

Ad= wound area on corresponding day.<\/p>\n

The mean percentage of wound contraction and standard error of mean were calculated in control ( Normal Saline), GTN and \u00a0GTN Ointment Base treated groups. The time required for epithelization was\u00a0 assessed in terms of days required for total fall of eschar with no trace of wound and full covering by glistening young epithelium.<\/p>\n

Statistical Analysis<\/strong><\/p>\n

The level of significance between individual group was analysed \u00a0using one way ANOVA. Data was expressed as mean \u00b1 SEM with a probability of\u00a0 p < 0.05 considered to be significant.<\/p>\n

After completion of epithelization, animals were followed up by standard procedures as outlined by CPCSEA guidelines and rehabilitated.<\/p>\n

Observations and Results<\/strong><\/p>\n

Table 2:\u00a0Percentage of wound contraction.<\/strong><\/p>\n\n\n\n\n\n\n\n
Group<\/strong><\/p>\n

 <\/td>\n

No of Animals<\/strong><\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 .Drugs<\/strong><\/td>\nMean Percentage Wound Contraction\u00a0 \u00b1\u00a0 .S.E.M.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0<\/strong><\/td>\n<\/tr>\n
<\/td>\n6th<\/sup> day.<\/strong><\/td>\n\u00a0\u00a0\u00a0 10th<\/sup> day<\/strong><\/td>\n14th<\/sup> day<\/strong><\/td>\n<\/tr>\n
1<\/strong><\/td>\n6<\/strong><\/td>\n\u00a0\u00a0 Control(Normal Saline)<\/td>\n\u00a0\u00a0\u00a0\u00a0 55.33\u00b16.04<\/td>\n\u00a0\u00a0 74.00\u00b11.63<\/td>\n90.33\u00b11.82<\/td>\n<\/tr>\n
2<\/strong><\/td>\n6<\/strong><\/td>\nGTN<\/td>\n\u00a0\u00a0 70.67\u00b1 2.04 *<\/sup><\/td>\n\u00a0\u00a0 81.00\u00b11.98\u00a0 *<\/td>\n90.33\u00b1 1.50<\/td>\n<\/tr>\n
3<\/strong><\/td>\n6<\/strong><\/td>\nOintment Base<\/td>\n\u00a0\u00a0\u00a0\u00a0 67.33\u00b18.04<\/td>\n\u00a0\u00a0\u00a0\u00a0 72.33\u00b17.42<\/td>\n94.6\u00b11.34<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Data is expressed as Mean \u00b1 S.E.M.\u00a0 p < 0.05\u00a0 is significant *=\u00a0\u00a0 p < 0.05 When\u00a0 compared\u00a0 with Control<\/p>\n

There was statistically significant difference in mean percentage of wound contraction of drug teated groups \u2013 GTN, when compared with Control\u00a0\u00a0 indicating GTN, has enhanced wound contraction than Control on 10th<\/sup> day.<\/p>\n

There was no statistically significant difference in mean percentage of wound contraction of drug treated groups \u2013 GTN, when compared with Control\u00a0\u00a0 indicating GTN, has not enhanced wound contraction than Control on 6th<\/sup> day and 14th<\/sup> day.<\/p>\n\n\n\n
\u00a0\"Graph<\/td>\n\n

Graph 1<\/strong><\/p>\n

Click here to View graph<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Period of Epithelization in Days<\/strong><\/p>\n\n\n\n\n\n\n
Group no.<\/strong><\/td>\nNo of Animals<\/strong><\/td>\nDrugs<\/strong><\/td>\nMean Period Of\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Epithelization<\/strong><\/td>\n<\/tr>\n
1.<\/strong><\/td>\n6<\/td>\nControl<\/td>\n19.00\u00b10.45<\/td>\n<\/tr>\n
2.<\/strong><\/td>\n6<\/td>\nGTN<\/p>\n

 <\/td>\n

17.83\u00b1 0.31<\/td>\n<\/tr>\n
3.<\/strong><\/td>\n6<\/td>\nOintment Base<\/td>\n19.00\u00b10.45<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Data is expressed as Mean \u00b1 S.E.M.\u00a0\u00a0\u00a0\u00a0 p < 0.05\u00a0 is significant\u00a0\u00a0 * = p < 0.05 compared with Control<\/p>\n

There was no statistically significant difference in means of period of epithelization with drug treated groups \u2013 GTN, GTN Ointment base, when compared to control indicating similar effect.<\/p>\n\n\n\n
\u00a0\"Graph<\/td>\n\n

Graph 2<\/strong><\/p>\n

Click here to View graph<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Discussion<\/strong><\/p>\n

GTN is used to treat chronic anal fissure. It acts by release of nitric oxide (NO), which increases concentration of Guanylyl cyclase. It increases cyclic GMP and brings out vasodilatation. GTN promotes wound healing by increasing blood supply and there by nourishment to the wound area.\u00a0 In addition, GTN activates cGMP, an intracellular intermediate, that inhibits the calcium activity. Calcium modulates cellular proliferation, modification, maturation of \u00a0keratinocytes and fibriblasts.11<\/sup><\/p>\n

GTN increases angiogenesis which is more vital. Wound contraction mediated by\u00a0 myofibroblasts\u00a0 is a secondary effect\u00a0 to\u00a0 angiogenesis. GTN application stimulates the production of organized collagen fibres and dampens inflammatory response by reducing\u00a0 number of\u00a0 polymorphonuclear cells. An NO inhibitor or NO synthase inhibitor can retard wound healing and NO donors have beneficial effects on wound healing,\u00a0 which confirms the role of\u00a0 NO in wound healing.11<\/sup><\/p>\n

GTN acts as an NO donor. NO plays an important role in modulating cytokines that initiate the process of inflammation after formation of wound. NO modulates inflammation induced oedema formation and it inhibits inflammatory cell infiltration into granulomas. It is vital for the activity of pro-angiogenic cytokines. It also important in the Vascular Endothelial Growth Factor (VEGF) dependent and independent angiogenesis. Angiogenesis in wound bed is important to maintain newly formed granulation tissue. Keratinocyte proliferation and wound re-epithelization is NO dependent, which is mediated by VEGF. It plays important role in collagen synthesis which is important in the proliferative phase of wound healing, as collagen gives strength to wound.12,13<\/sup>. In\u00a0 a previous study it has been observed that NO has promoted re- epithelization in healing of wounds.6,14<\/sup>. An NO inhibitors or iNOS\u00a0\u00a0 inhibitor can retard wound healing by decreasing proliferation of keratinocytes and delays healing.15,16 <\/sup><\/p>\n

The present study was undertaken to determine if GTN possesses wound healing potential.\u00a0 \u00a0The property of GTN regarding wound healing was compared with control group. GTN has shown increase in mean percentage of wound contraction on 10th<\/sup> day after wounding. Percentage of wound contraction of Control group (74.00\u00b11.63) and\u00a0 that of GTN group is (81.00\u00b11.98).\u00a0 There was statistically significant difference in mean percentage of wound contraction and GTN was better than control group. There was no statistically significant difference in period of\u00a0 epithelization\u00a0 when compared with control.<\/p>\n

Thus from\u00a0 present study it seems that apart from spasmolytic activity Glyceryl trinitrate has wound healing property\u00a0 as it has enhanced mean percentage of wound contraction.<\/p>\n

Conclusion<\/strong><\/p>\n

Apart from the spasmolytic activity of Glyceryl trinitrate, wound healing property has been demonstrated in our study, as it has enhanced mean percentage of wound contraction.<\/p>\n

Acknowledgement<\/strong><\/p>\n

Authors acknowledge to Mrs Gore, Associate Professor, Department of Preventive and Social Medicine, Bharati Medical college and Hospital, Sangli for her valuable support in statistical analysis.<\/p>\n

Conflict of Interest<\/strong><\/p>\n

No conflict of Interest<\/p>\n

Funding Source<\/b><\/p>\n

No funding was required<\/p>\n

References<\/strong><\/p>\n

    \n
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  7. Franceschilli L ,Luca E, Milito G,Gaspari AL.Role of 0.4% glyceryl trinitrate ointment after haemorrhoidectomy. Int J Colorectal Dis.2012 Aug; 1544-51.<\/li>\n
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  11. Tripathi KD. Antianginal & other Antieschemic Drugs.In: Tripathi KD, editor.Essentials of Medical Pharmacology. New Delhi: Jaypee Brothers Medical Publishers(p) Ltd; 2013. p.539-57.<\/li>\n
  12. Chong DY, Michel T. Pharmacology of vascular tone. In: Golan DE, Tashjian AH, Armastrong\u00a0 EJ, Armstrong AW, editor.Principles of\u00a0 pharmacology. Phildelphia: Lippincott Williams & Wilkins publisher; 2008. P. 367- 81.<\/li>\n
  13. Katzung BG, Vasodilators\u00a0 &\u00a0 The Treatment of\u00a0 Angina\u00a0 Pectoris. In: Katzung BG, Trevor AJ, editor. Basic &\u00a0 Clinical Pharmacology. New Delhi : McGraw Hill Education(India). Publisher; 2015. P.191- 207.<\/li>\n
  14. Stallmeyer B,Anhold M,Wetzler C,Kahlina K,Pfelschifter J, Frank S.Regulation of Enos in normal and diabetis- impaired skin repair: implications for tissue regeneration. Nitric Oxide. 2002; 6: 168-77.<\/li>\n
  15. Opal SM,DePalo VA. Anti- inflammatory cytokines.Chest 2000; 117: 1162-72.<\/li>\n
  16. Zuh H, Ka B,Murad F. Nitric Oxide accelerates the recovery from burn wounds. World J Surg .2007;31: 624-31.<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    Introduction Wound healing is a complex, delicate process which consists […]<\/p>\n","protected":false},"author":8,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[73],"tags":[],"class_list":["post-29372","post","type-post","status-publish","format-standard","hentry","category-vol12no4"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/29372","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=29372"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/29372\/revisions"}],"predecessor-version":[{"id":31799,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/29372\/revisions\/31799"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=29372"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=29372"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=29372"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}