Effectiveness of IV Vitamin C in Reducing Oxidative Stress Associated With Free Flap Surgery

Oxidative Stress Clinical Trial

Official title:

The Efficacy of Perioperative Parenteral Ascorbic Acid in Reducing Oxidative Stress Among Patients Undergoing Free Flap Reconstructive Surgery: A Prospective Multicenter Randomized Controlled Pilot Trial

Clinical Trial Details — Status: Recruiting

Administrative data

• NCT number: NCT05327348
• Other study ID #: MECID No,UMMC: 202086-8956
• Secondary ID: PV042-2021NMRR-2
• Status: Recruiting
• Phase: Phase 3
• Start date: September 25, 2022
• Est. completion date: July 31, 2023

Study information

• Verified date: November 2022
• Source: University of Malaya
• Contact: Raymond Yii Shi Liang, MBBS
• Phone: +60379494422
• Email: raymond.yii[@]ummc.edu.my
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial Summary

Ischemia and reperfusion injury during free flap reconstructive surgery creates a state of increased oxidative stress that can adversely affect the flap outcomes. Ascorbic acid (AA) had been proven to have beneficial effect on end-organ protection and flap survival from ischemia-reperfusion injury via its antioxidant properties. The investigators hypothesise that perioperative parenteral ascorbic acid treatment may reduce oxidative stress among participants undergoing free flap reconstructive surgery along with reduction in inflammatory markers, improved rate of flap viability and wound healing at both donor and recipient sites.

Description:

Temporary cessation of blood supply to the flap tissues from clamping of donor vascular pedicle (ischemia) followed by restoration of flap tissue perfusion (reperfusion) after micro-anastomosis are inevitable in any free flap surgery. In combination, the ischemia and reperfusion process during free flap tissue transfers induce a state of increased oxidative stress, which may lead to complications such as flap failure and non-healing wounds at either donor or recipient sites. The negative impacts of which include additional wounds from flap loss, higher costs and increased duration of hospital stay. Previous studies had demonstrated the beneficial effects of ascorbic acid in end-organ protection against ischemia and reperfusion injury. In addition, parenteral ascorbic acid has been shown to be remarkably safe even at high dose in both clinical and nonclinical models. Nonetheless, the data on efficacy of ascorbic acid in free flap survival in human is very limited. The aims of this prospective, multicentre, double-blind, randomized, placebo-controlled pilot study are to measure the extent of oxidative stress in participants undergoing free flap reconstructive surgery before and after administration of parenteral ascorbic acid; and to evaluate its efficacy on modulation of inflammation, flap viability and wound healing. Eligible participants will be randomized to receive 1 gram of parenteral ascorbic acid and 0.9% normal saline (as placebo) 8 hourly for 7 days (from pre-operative day 2 until post-operative day 5). Blood sampling will be performed on day 0 (pre-operative), day 3 (post-operative day 1) and day 5 (post-operative day 3) of intravenous ascorbic acid or placebo infusion for measurement of i) oxidative stress biomarkers, including isoprostane level, gene expression of glutamate-cystein ligase (GCL) and total glutathione level) ii) inflammatory markers, including leucocytes count and gene expression of TNF-α and IL-1. Post-operative outcomes of free flap surgery, up to post-operative 14 days, including flap viability, wound healing at both donor and recipient sites and duration of ICU and hospital stay will be evaluated. The investigators estimate that a total sample of 28 participants (14 on each arm) will be necessary for 80% power to detect a 33% oxidative stress reduction with medium effect size (0.5) at 5% level of significance (α) between treatment (intravenous ascorbic acid) and placebo group (0.9% normal saline). A total of 34 participants are required to account for 20% of dropouts. Primary analysis of this study utilizes an intention-to-treat approach and includes all randomized participants undergoing elective free flap reconstructive surgery. The mean difference between the baseline (pre-operative) and post-operative oxidative stress and inflammatory levels will be analyzed and compared between the intravenous ascorbic acid and placebo group using analysis of variance (ANOVA) for all normally distributed dataset whilst the non-parametric Kruskal-Wallis test is used, if otherwise. The effect size of such difference will be determined and compared. Subsequently, correlation between reduction of oxidative stress and post-operative flap outcomes in the intravenous ascorbic acid group will be evaluated. The secondary outcomes such as flap viability (percentage of flap necrosis), wound healing at both recipient and donor sites (percentage of wound dehiscence and percentage of skin graft failure to take/loss), duration of hospital and ICU stay and wound infection rate will be presented as mean with standard deviation (SD) or median with interquartile range (IQR) based on their normality distribution and are compared with Student's t-test or Mann-Whitney U test.

Recruitment information / eligibility

• Status: Recruiting
• Enrollment: 34
• Est. completion date: July 31, 2023
• Est. primary completion date: July 31, 2023
• Accepts healthy volunteers: No
• Gender: All
• Age group: 18 Years and older

Eligibility

Inclusion Criteria:

• Adults (age 18 years and older, male or female) who are planned for elective free flap reconstructive surgery.

Exclusion Criteria:

• Hypersensitivity to vitamin C
• Oliguria (urine output <400mL/day) or anuria (urine output <100mL/day)
• Renal failure (serum creatinine level =175.0 %mol/L)
• Hemodialysis
• Renal calculi
• Thalassemia
• Glucose-6-phosphate dehydrogenase (G6PD) de?ciency
• Unfit for surgery
• Pregnancy or lactating
• Hemochromatosis
• Hyperoxaluria

Related Conditions & MeSH terms

• Ischemia
• Ischemia-reperfusion Injury
• Oxidative Stress
• Reperfusion Injury

Intervention

Drug:
• Intravenous Ascorbate
Intravenous ascorbic acid 1 gram 8 hourly (3 grams per day) over 15 minutes for 7 days since pre-operative day 1 until post-operative day 5.
• Normal Saline 10 mL Injection
Intravenous 0.9% normal saline 8 hourly bolus infusion over 15 minutes for 7 days since pre-operative day 1 until post-operative day 5.

Locations

Country	Name	City	State
Malaysia	Hospital Kuala Lumpur	Kuala Lumpur
Malaysia	University of Malaya Medical Centre	Kuala Lumpur
Malaysia	Hospital Universiti Sains Malaysia	Kubang Kerian	Kelantan

Sponsors (1)

Lead Sponsor	Collaborator
University of Malaya

Country where clinical trial is conducted

Malaysia, 

References & Publications (13)

Azari O, Kheirandish R, Azizi S, Farajli Abbasi M, Ghahramani Gareh Chaman S, Bidi M. Protective Effects of Hydrocortisone, Vitamin C and E Alone or in Combination against Renal Ischemia-Reperfusion Injury in Rat. Iran J Pathol. 2015 Fall;10(4):272-80. — View Citation

Ballestín A, Casado JG, Abellán E, Vela FJ, Álvarez V, Usón A, López E, Marinaro F, Blázquez R, Sánchez-Margallo FM. Ischemia-reperfusion injury in a rat microvascular skin free flap model: A histological, genetic, and blood flow study. PLoS One. 2018 Dec 27;13(12):e0209624. doi: 10.1371/journal.pone.0209624. eCollection 2018. — View Citation

Fowler AA 3rd, Syed AA, Knowlson S, Sculthorpe R, Farthing D, DeWilde C, Farthing CA, Larus TL, Martin E, Brophy DF, Gupta S; Medical Respiratory Intensive Care Unit Nursing, Fisher BJ, Natarajan R. Phase I safety trial of intravenous ascorbic acid in patients with severe sepsis. J Transl Med. 2014 Jan 31;12:32. doi: 10.1186/1479-5876-12-32. — View Citation

Hoffer LJ, Robitaille L, Zakarian R, Melnychuk D, Kavan P, Agulnik J, Cohen V, Small D, Miller WH Jr. High-dose intravenous vitamin C combined with cytotoxic chemotherapy in patients with advanced cancer: a phase I-II clinical trial. PLoS One. 2015 Apr 7;10(4):e0120228. doi: 10.1371/journal.pone.0120228. eCollection 2015. — View Citation

Lee WY, Lee JS, Lee SM. Protective effects of combined ischemic preconditioning and ascorbic acid on mitochondrial injury in hepatic ischemia/reperfusion. J Surg Res. 2007 Sep;142(1):45-52. Epub 2007 Jun 7. — View Citation

Padayatty SJ, Sun AY, Chen Q, Espey MG, Drisko J, Levine M. Vitamin C: intravenous use by complementary and alternative medicine practitioners and adverse effects. PLoS One. 2010 Jul 7;5(7):e11414. doi: 10.1371/journal.pone.0011414. — View Citation

Schäfer M, Werner S. Oxidative stress in normal and impaired wound repair. Pharmacol Res. 2008 Aug;58(2):165-71. doi: 10.1016/j.phrs.2008.06.004. Epub 2008 Jun 19. Review. — View Citation

Siemionow M, Arslan E. Ischemia/reperfusion injury: a review in relation to free tissue transfers. Microsurgery. 2004;24(6):468-75. Review. — View Citation

Stepanovs J, Ozolina A, Rovite V, Mamaja B, Vanags I. Factors Affecting the Risk of Free Flap Failure in Microvascular Surgery. Proc Latv Acad Sci Sect B Nat Exact, Appl Sci. 2016;70(6):356-364. doi:10.1515/prolas-2016-0039.

Stephenson CM, Levin RD, Spector T, Lis CG. Phase I clinical trial to evaluate the safety, tolerability, and pharmacokinetics of high-dose intravenous ascorbic acid in patients with advanced cancer. Cancer Chemother Pharmacol. 2013 Jul;72(1):139-46. doi: 10.1007/s00280-013-2179-9. Epub 2013 May 14. — View Citation

Tsai MS, Huang CH, Tsai CY, Chen HW, Lee HC, Cheng HJ, Hsu CY, Wang TD, Chang WT, Chen WJ. Ascorbic acid mitigates the myocardial injury after cardiac arrest and electrical shock. Intensive Care Med. 2011 Dec;37(12):2033-40. doi: 10.1007/s00134-011-2362-6. Epub 2011 Sep 28. — View Citation

Wang ZJ, Hu WK, Liu YY, Shi DM, Cheng WJ, Guo YH, Yang Q, Zhao YX, Zhou YJ. The effect of intravenous vitamin C infusion on periprocedural myocardial injury for patients undergoing elective percutaneous coronary intervention. Can J Cardiol. 2014 Jan;30(1):96-101. doi: 10.1016/j.cjca.2013.08.018. — View Citation

Zaccaria A, Weinzweig N, Yoshitake M, Matsuda T, Cohen M. Vitamin C reduces ischemia-reperfusion injury in a rat epigastric island skin flap model. Ann Plast Surg. 1994 Dec;33(6):620-3. — View Citation

* Note: There are 13 references in all — Click here to view all references

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Oxidative stress biomarker: Plasma isoprostane level	Plasma isoprostane level will be analyzed using a competitive ELISA assay and expressed in unit of pg/ml.	Change from preoperative baseline level at day 5 of infusion (post-operative)
Primary	Oxidative stress biomarker: Gene expression of glutamate-cystein ligase (GCL)	The expression of glutamate cysteine ligase (GCL) will be quantitated using real time quantitative polymerase chain reaction (qRT-CR). The differences in gene expression, expressed as fold-change, will be calculated using the 2 -D?Ct algorithm where GAPDH will be used as the housekeeping gene.	Change from preoperative baseline level at day 5 of infusion (post-operative)
Primary	Oxidative stress biomarker: Total glutathione level	Total glutathione level (GSSG + GSH) will be determined using a Glutathione Assay Kit, which will be expressed in unit of µM.	Change from preoperative baseline level at day 5 of infusion (post-operative)
Secondary	Inflammatory biomarkers levels: Leucocytes counts	Leucocytes count will be measured through flow cytometry White blood cells Differential Fluorescence (WDF) scattergram using Sysmex XN-10 (Sysmex Corporation TM , Kobe, Japan) and expressed in unit of X10^9/L.	Change from preoperative baseline level at day 5 of infusion (post-operative)
Secondary	Inflammatory biomarkers levels: Gene expression of TNF-a	The expression of TNF-a will be quantitated using real time quantitative polymerase chain reaction (qRT-CR). The differences in gene expression, expressed as fold-change, will be calculated using the 2 -D?Ct algorithm where GAPDH will be used as the housekeeping gene.	Change from preoperative baseline level at day 5 of infusion (post-operative)
Secondary	Inflammatory biomarkers levels: Gene expression of IL-1	The expression of IL-1 will be quantitated using real time quantitative polymerase chain reaction (qRT-CR). The differences in gene expression, expressed as fold-change, will be calculated using the 2 -D?Ct algorithm where GAPDH will be used as the housekeeping gene.	Change from preoperative baseline level at day 5 of infusion (post-operative)
Secondary	Post-operative outcomes: Flap viability	Flap viability is the survival of flap without tissue loss. For flap viability assessment, total flap loss is defined as complete necrosis (death) of flap (100% flap loss), whilst partial flap loss refers to incomplete necrosis of flap. By using simple tracing technique, the percentage of flap necrosis is calculated as: [Necrotic flap surface area (cm^2)/Total flap surface area (cm^2)] × 100%	From post-operative day 1 until day 14 (2 weeks)
Secondary	Post-operative outcomes: Wound dehiscence	Surgical wound dehiscence is defined as separation of the margins of a closed surgical incision that has been made in skin, with or without exposure or protrusion of underlying tissue, organs or implants. Wound dehiscence is assessed upon suture removal. It is categorized as either partial (1-99%) or complete (100%), depending on the depth/thickness of skin layers involvement. Partial wound dehiscence is defined as separation of wound edge up to level of epidermis and dermis layer of sutured wounds, whereas complete wound dehiscence refers to complete separation of wound edge involving the full thickness of skin. The percentage of wound dehiscence in relative to the total wound size is calculated.	From post-operative day 1 until day 14 (2 weeks)
Secondary	Post-operative outcomes: Graft loss	Skin graft recipient area is inspected at post-operative day 5 for assessment of graft take. Graft loss is categorized as partial (1-99%) or complete (100%). The percentage of skin graft failure to take/loss is calculated as: [Graft failure to take or loss surface area (cm^2)/Total skin graft area surface area (cm^2)] X 100%	From post-operative day 1 until day 14 (2 weeks)
Secondary	Post-operative outcomes: Wound infection	Wound infection is defined by presence of clinical sign and symptoms of infection associated with wound, including: i) Local signs: Erythema - localized or spreading (cellulitis); Pus/purulent or haemopurulent exudate; Abscess; Swelling/induration; Local warmth; Malodour; Crepitus; Dehiscence; Unexpected pain or tenderness.; ii) Systemic signs: Malaise; Loss of appetite; Pyrexia or hypothermia; Tachycardia; Tachypnoea; Elevated C-reactive protein (CRP); Elevated or suppressed white blood cell count; Sepsis or Septic Shock.	From post-operative day 1 until day 14 (2 weeks)