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Spurgeon raj Jalem1 & Purimitla Usharani2,*

1Department of Emergency medicine and critical care, Bliss Hospital, Murad Nagar Road, Mehidipatnam, Hyderabad - 500028,

Telangana, India; 2Department of Microbiology, Dr. Patnam Mahender Reddy Institute of Medical Sciences, Chevella, Ranga Reddy,

Telangana, India; *Corresponding author

Abstract:

Wound healing is a complex process involving multiple stages to restore the integrity and function of damaged tissues. Researchers are exploring the therapeutic potential of HOCl in treating infections and inflammatory conditions, given its efficacy in killing pathogens and modulating immune responses. The present study is a prospective study aimed to know the wound healing nature of HOCl on open wounds in patients attending out-patient department. The study was conducted for a period of one year Jan 2023 to Jan 2024 at Bliss Hospital, Hyderabad, Telangana district, India. Hypochlorous acid (Vida solutions, Pvt.Ltd) was used to irrigate the open wound. Among the 10 reported cases, three were females and seven were males. Most of the cases were reported from middle-age and old-age persons except one paediatric case of age 3 years a female girl. A male patient of age 72 years with co-morbidities diabetes and hypertension suffering with chronic wound on left leg for a period of 35 years because of thorn prick injury. The】 complete wound healing took nearly 15 weeks in this patient. The wound healing takes place 8 to 15 weeks maximum in all the 10 cases and the above findings concluded that, these features lead to a stabilized HOCl solution as an ideal wound care agent.

Keywords: Hypochlorous acid, chronic wound, infection, co-morbidities

Serhan Sakarya, MD; Necati Gunay, MS; Meltem Karakulak, MS; Barcin Ozturk, MD; Bulent Ertugrul, MD Wounds. 2014;26(12):342-350.

Abstract and Introduction

Abstract

Introduction Chronic wounds and the infections associated with them are responsible for a considerable escalation in morbidity and the cost of health care. Infection and cellular activation and the relation between cells are 2 critical factors in wound healing.

Since chronic wounds offer ideal conditions for infection and biofilm production, good wound care strategies are critical for wound healing. Topical antiseptics in chronic wounds remain in widespread use today. These antiseptics are successful in microbial eradication, but their cytotoxcity is a controversial issue in wound healing.

Objective The aim of this study was to investigate the effect of stabilized hypochlorous acid solution (HOCl) on killing rate, biofilm formation, antimicrobial activity within biofilm against frequently isolated microorganisms and migration rate of wounded fibroblasts

and keratinocytes.

Materials and Methods. Minimal bactericidal concentration of stabilized HOCl solution for all standard microorganisms was 1/64 dilution and for clinical isolates it ranged from 1/32 to 1/64 dilutions.

Results All microorganisms were killed within 0 minutes and accurate killing time was 12 seconds. The effective dose for biofilm impairment for standard microorganisms and clinical isolates ranged from 1/32 to 1/16. Microbicidal effects within the biofilm and

antibiofilm concentration was the same for each microorganism.

Conclusion. The stabilized HOCl solution had dose-dependent favorable effects on fibroblast and keratinocyte migration

compared to povidone iodine and media alone. These features lead to a stabilized HOCl solution as an ideal wound care agent.

Introduction

Wound healing is a sequence of complex and well-orchestrated events. Although the precise mechanism of wound healing is not fully understood, 3 interrelated phases—inflammation, migration, and remodeling—require coordinated activity for successful wound healing, which is a progressive series of events facilitated by platelets, leukocytes, fibroblasts, and keratinocytes. Platelets facilitate homeostasis and the release of growth factors, then leukocytes participate in the inflammatory process. Fibroblasts and keratinocytes have a critical role in wound healing by enhancing reepithelization and the remodeling of the extracellular matrix (ECM).[1–3] Most chronic wounds are related to diabetes mellitus, venous stasis, peripheral vascular diseases, and pressure ulcerations. An open wound is a favorable niche for bacterial colonization and infection. Infection in chronic wounds starts with contamination, then colonization and critical colonization take place before an infection forms. Biofilm formation is now recognized as a serious problem in chronic wound infections.[4] Biofilm is a complex structure of microorganisms that generate a protective shell, allowing bacteria to collect and proliferate.[5] Most of the microorganisms that form biofilms can also be found growing in microbial infections. The same species of bacteria have significant differences in existence that range either free floating and living within the biofilm. The biofilm structure of microorganisms renders phagocytosis difficult,[6] increases resistance to antibiotics,[7] and adheres to unfavorable niches such as chronic wounds.[8] One of the remarkable features of the immune system against invading pathogens is its ability to generate an effective and rapid response by developing a group of highly reactive chemicals, such as reactive oxygen species (ROS). The mitochondrial membrane-bound enzyme nicotinamide adenine dinucleotide phosphate-oxidase (NADPH) is a primary enzyme responsible for ROS production.[9] During the activation of neutrophils, respiratory bursts generate hydrogen peroxide (H2O2) and the activated granule enzyme myeloperoxidase converts H2O2 to hypochlorous acid (HOCl) in the presence of Cl- and H+.[10] Hypochlorous acid leads to cell death by the oxidation of sulfhydryl enzymes and amino acids, ring chlorination of amino acids, loss of intracellular contents, decreased uptake of nutrients, inhibition of protein synthesis, decreased oxygen uptake, oxidation of respiratory components, decreased adenosine triphosphate production, breaks in DNA, and depressed DNA synthesis.[11–15] Hypochlorous acid is highly active against all bacterial, viral, and fungal human pathogens[16] and a small amount of HOCl can kill spore-forming and non-spore bacteria in a short time period.[17,18] Since most of the etiologic factors in chronic wound infections are forming biofilm,[19] and most of the topical antiseptics impair wound healing with their cytotoxic effect, therapeutic strategies against biofilm with high microbial eradication and good wound healing effects will decrease the morbidity and mortality rates of patients and reduce the economic burden. The aim of this study was to investigate the effect of stabilized HOCl solution on microbial and biofilm eradication, antimicrobial activity within biofilm against frequently isolated microorganisms, and migration rate of wounded fibroblasts and keratinocytes.

Craig Meyers¹, Janice Milici¹, Lori Robins², Luis Contreras², Jeffrey Williams³, Richard Robison4 ¹Department of Microbiology and Immunology, Penn State College of Medicine, Hershey, Pennsylvania 17033, USA; ²Physical Science Division, School of STEM, University of Washington Bothell, Bothell, Washington 98011, USA; ³Briotech Inc., Woodinville, Washington 98072, USA; 4Department of Microbiology and Molecular Biology, Brigham Young University, Provo, Utah 84602, USA

Tomislav Bulum 1,2,* , Tamara Poljiˇcanin 3 , Anica Badanjak 4 , Jelena Držiˇc 4 and Željko Metelko 4

1 Clinical Hospital Merkur, University Clinic for Diabetes, Endocrinology, and Metabolic Diseases Vuk Vrhovac, Dugi dol 4a, 10000 Zagreb, Croatia

2 School of Medicine, University of Zagreb, Šalata 3, 10000 Zagreb, Croatia

3 Zagreb County Health Center, Ljudevita Gaja 37, 10430 Samobor, Croatia

4 Polyclinic for Physical Medicine and Rehabilitation with Physical Therapy, Vascular Surgery, Neurology, Endocrinology, and Diabetology, Kalinovica 3, 10000 Zagreb, Croatia

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Abstract: (1) Background: Diabetes-related foot ulcers (DFUs) are a severe complication of diabetes mellitus (DM), with a five-year mortality rate of around 40%. Our study aimed to explore the effects of transcutaneous application of carbon dioxide (CO2 therapy) on DFU healing and recurrence rate, as well as diabetic polyneuropathy. (2) Methods: Adults with at least one chronic DFU were invited to undergo 20 50-min-long CO2 therapies within 4 weeks. After the completion of the last CO2 therapy, the effect of the therapies on wound healing and diabetic polyneuropathy was assessed, and 1 year later, we evaluated the incidence rate of DFU recurrence. (3) Results: Thirty-five subjects with DM and forty DFUs (ischemic: 15, neuropathic: 8, neuroischemic: 17) participated in our trial. After 4 weeks, 67.5% of all DFUs healed, whereas the reduction of the surface area of the unhealed wounds (74.0% ± 22.3%) was statistically significant. The restoration of protective sensations was also statistically significant. All unhealed wounds received standard care and healed within 2 additional weeks. The recurrence rate after 1 year was 17.5%. None of the patients required antibiotic treatment, hospitalization, or amputation. (4) Conclusion: CO2 therapy is a promising therapeutic intervention for treating DFUs and improving diabetic polyneuropathy.

Keywords: diabetic foot; diabetes-related foot ulcer; CO2 therapy; transcutaneous CO2 application; wound healing; distal symmetrical polyneuropathy; loss of protective sensation

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