ANTIBACTERIAL ACTIVITIES OF CRUDE Curcuma longa EXTRACT MEDIATED SILVER NANO PARTICLES AGAINST ISOLATES FROM DIABETIC PATIENTS WITH FOOT INFECTIONS

ABSTRACT

Curcuma longa are traditionally used for the treatment of ulcers, hepatic disorder, wound healing and boost glucose control. This study assessed the phytochemical compositions (quantitative and qualitative), antibacterial activities of C. longa crude extracts, synthesis and characterisation of silver nanoparticles, antibacterial activity of extract-mediated silver nanoparticles and wound activity of EeaAgNPs. The antibacterial activity of crude extracts and extract mediated silver nanoparticles (AgNPs) of C. longa rhizomes were evaluated against isolates from patients with diabetic foot infection using agar well diffusion method. Quantitative determination of phytoconstituents revealed a significant amount of phytates (6577.9 mg/100 g), cyanides (2741.8 mg/100 g) and saponins (618.0 mg/100 g). Cold maceration of rhizome with 70% ethanol yielded a crude extract (E). Successive partitioning of extract E with chloroform and ethyl acetate yielded chloroform (Ec), ethyl acetate (Eea) soluble fractions, as well as aqueous residual fraction (Eaq). Qualitative screening of the extract and fractions revealed the presence of flavonoids, reducing sugars, anthraquinones, tannins and saponins. The extract and its fractions at 100 mg/ml were inactive on Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Streptococcus pyogenes and Staphylococcus aureus while, fraction Eea at 200mg/ml was active on P. aeruginosa, E. coli and S. pyogenes. The wavelength of E- AgNPs, Ec-AgNPs, Eea-AgNPs, and Eaq-AgNPs were 405 nm, 406 nm, 409 nm and 410 nm respectively. The FTIR indicated the presence of aromatic, alkanes, alkynes, alkenes and carboxylic functional groups while the SEM micrograph of Eea-AgNPs revealed clustered rod-like morphology. The highest XRD peak was at 2ϴ (340). The Eea-AgNPs at 200 mg/ml was active on S. pyogenes, P. aeruginosa, E. coli and K. pneumoniae with zones of inhibition of 7 ± 1.7 mm, 10 ± 0.7 mm, 11 ± 1.1 mm and 14 ± 0.5 mm respectively. The MIC of Eea-AgNPs against test isolates was at 12.5 mg/ml. The extract was bacteriostatic on the test isolates. There was significant (P<0.05) wound closure observed in rats (groups 1 to 6) topically treated with Eea-AgNPs ointment from Day 0 (1.24 ± 0.00 mm to 1.29 ± 0.19 mm) to Day 14 (0.4 ± 0.1 mm to 0.73 ± 0.00 mm) compared to group 7 (Diabetes + Wound only) with (1.23 ± 0.00 to 1.1 ± 0.3 mm). Histology of the treated rats indicated wound healing characterized with collagens, fibroblasts, inflammatory cells, new blood vessels, granulation tissues and complete epithelialization. The application of ointment on rats produced no allergic reactions, rashes and other forms of skin irritation. These findings showed the potentials of C. longa as a safe therapeutic agent to treat and heal infected ulcer.

CHAPTER ONE

1.0     INTRODUCTION

1.1       Background to the Study

Diabetes mellitus (DM) is a complex metabolic disorder characterized by hyperglycemia, pancreatic beta (ß) cells dysfunction, and abnormal lipid profile that result from metabolic deregulations, impaired insulin secretion and action, and inappropriate consumption of glucose (Iftikhar et al., 2020). Diabetes foot infection is ulceration of tissues of the foot associated with neuropathy disease in a person with diabetes mellitus (Netten et al., 2019).

Diabetic foot infection (DFI) is one of the diabetic complications associated with major morbidity, mortality, and reduced quality of life and is the most serious complication of diabetes mellitus and  one of the main problems in health systems and a global public health threat that has increased dramatically over the past two decades (Abdissa et al., 2020). According to epidemiological studies, the number of patients with diabetes mellitus increased from about 30 million cases in 1985, 177 million in 2000, 285 million in 2010, and it is estimated that if the situation continues, more than 360 million people by 2030 will have diabetes mellitus (Yazdanpanah et al., 2015; Alhubail et al., 2020; Sharma et al., 2021).

Global prevalence of diabetes is high and still on the rise. The prevalence’s in the world, Africa, and Nigeria stand at 8.8 %, 3.2 %, and 4.6 %, respectively. An increase in the prevalence of diabetes is accompanied by an increase in its complications such as foot ulcers and lower extremity amputations, in that, the lifetime risk of a person with diabetes developing a foot ulcer is 25 % (Ambrose and Christopher, 2019). The risk for lower extremity amputation is 15 to 40 times higher in people with diabetes than people without diabetes. The complications of diabetes result in reduced quality of life, incapacity and death. With regard to diabetic foot ulcers, 12 % of all hospitalized diabetic patients in Africa have foot ulceration. Research indicates that diabetes patients with foot ulcers encounter stigma, loss of social role, social isolation, and unemployment. Diabetic foot ulcer is a costly and debilitative disease with severe consequences in diabetic patients. Also, mortality after lower extremity amputations in diabetes patients varies from 39 % to 80 % in 5 years (Ambrose and Christopher, 2019). More than half of all non-traumatic lower limb amputations are due to diabetes. Limb amputation causes distortion of body image, increase in dependency, loss of productivity, and increase in costs of treating diabetic foot ulcers (Ambrose and Christopher, 2019).

Risk factors for the development of DFIs include neuropathy, peripheral vascular disease, and poor glycemic control. In sensory neuropathy, there is diminished perception of pain and temperature. Autonomic neuropathy can cause diminished sweat secretion resulting in dry, cracked skin that facilitates the entry of microorganisms to the deeper skin structures. In addition, motor neuropathy can lead to foot deformities, which lead to pressure-induced soft tissue damage. Peripheral artery disease can impair blood flow necessary for healing of ulcers and infections. Hyperglycemia impairs neutrophil function and reduces host defenses. Trauma in patients with one or more of these risk factors precipitates the development of wounds that can be slow to heal and predispose to secondary infection (Vasanthan et al., 2018).

Diabetic foot infections are frequent clinical problems. Infection in foot wounds may be defined clinically by the presence of inflammation or purulence, and then classified by severity. Many organisms, alone or in combinations, can cause DFI, but Gram-positive cocci, especially staphylococci, are the most common (Vasanthan et al., 2018). The impaired micro-vascular circulation in patients with diabetic foot limits the access of phagocytes   favoring   development   of   infection.   Escherichia   coli,   Proteus   spp.

Pseudomonas spp., Staphylococcus aureus and Enterococcus spp. are the most frequent pathogens contributing to methicillin-resistant Staphylococcus aureus (MRSA) has been commonly isolated from 10 – 40 % of the diabetic progressive and widespread tissue destruction. The increasing association of multi-drug resistant (MDR) pathogens with diabetic foot ulcers further compounds the challenge faced by the physician or the surgeon in treating diabetic ulcers without resorting to amputation (Gunasekaran, 2017).

The known sore infection is predictive of poor ulcer repair and amputation. Proper diagnosis of infection and antibiotic therapy in diabetic foot infections is necessary to ameliorate the yields, because wrong application of antibiotics can lead to resistance and side effects. Three principal factors, such as particular agents, route of administration and duration of remedy may reduce efficacy on the antibiotic therapy of Diabetic foot infection (Abolghasemi and Mesri, 2019).

Curcuma longa (commonly known as turmeric) is also known as Atale pupa by Yorubas, Gangamau by the Hausas, Ohu boboch in Igbo, Girgir by the Tivs. Turmeric is an herbaceous plant of the family Zingiberaceae that has been considered an important therapeutic agent in Indian and Chinese traditional medicine, it is mainly cultivated in tropical and subtropical regions. Turmeric contains curcumin which has therapeutic potential against neurodegenerative disorders, cardiovascular diseases, wound healing ability, hepatic damage, renal diseases and diabetes mellitus (Rashid et al., 2017; Rivera- Mancia et al., 2018).

Local medicinal uses of turmeric include the use of its herbal decoction with traditional distilled gin called “ogogoro” and it is consumed with the claim that it cures ailments like diabetes, ulcers, hepatic disorders and cough. The rhizome is also used as anti- inflammatory therapy for wounds, digestive disorders and jaundice in babies (Oghenejobo et al., 2017).

The identification of curcumin as the main constituent of turmeric has led to its multiple usage. Pharmacological activities of curcumin (the main active principle in turmeric) include antimicrobial, anti-diabetic, anti-inflammatory, and antioxidant More excitingly, when combined with other drugs, curcumin has been found to enhance the effects of antibacterial, antifungal, anticancer, and antioxidant activities, Curcumin usually exhibit low to no toxicity at the active doses (Sin-Yeang et al., 2016). The range of beneficial effects of curcumin in diabetes mellitus and its complications has been attributed to its ability to interact with many key molecules and pathways involved in the pathophysiology of this disease (Rivera-Mancia et al., 2018).

In order to enhance the efficacy of Curcuma longa extract the need to introduce a nanotechnology system was adapted which is aimed at developing an environmentally friendly and cost-effective approach to synthesise green silver nanoparticles (AgNPs) from silver precursors. This was accomplish using various extracts of turmeric powder, in which the plant biomaterials were used as  a reducing as well as capping agent (Alsammarraie et al., 2020).

Nanotechnology can be defined as the formation, development, enhancement and exploration of nano sized materials having size range of (1 – 100 nm) that confers them their unique physicochemical properties. It works with the substance which has specific properties such as physical, chemical and biological (Garg and Garg, 2018). Thus, nanotechnology offers an alternative to overcome infectious diseases through the use of antimicrobial nanomaterials (Hernandez et al., 2017).  Silver nanoparticles (AgNPs) is one of the most used nanomaterials in medical products such as bandages, wound dressings, catheters and textiles due to their excellent microbicidal activity against wild and nosocomial strains of MDR microorganisms (Salomoni et al., 2017).

The use of nanotechnology for phytotherapy or treatment of various diseases by herbal medicines, including herbal drug delivery where current and emerging nanotechnologies could enable entirely novel classes of therapeutics has been reported. Interestingly, pharmaceutical sciences are using nanoparticles to reduce toxicity and side effects of drugs.  The  biologically  synthesized  nanoparticles  with  plant  products  have  better chemotherapeutic effects against microbial diseases (Divakaran et al., 2018). Biosynthesis of AgNPs can be accomplished by physical, chemical, and green synthesis; however,  synthesis  via  biological  precursors  has  shown  remarkable  outcomes.  In available reported data, these entities are used as reducing agents where the synthesized nanoparticles  are  characterized  by  ultraviolet-visible  and  Fourier-transform  infrared spectra and X-ray diffraction, scanning electron and transmission electron microscopy (Ahmad et al., 2020).

1.2       Statement of the Research Problem

Diabetes foot ulcers and infection lead to substantial morbidity and mortality, pains, skin discoloration, abnormal leucocyte function, and deformity which may lead to partial or whole leg amputation, resulting in severe disability, reduced quality of life and high health cost (Nicolas et al., 2017; Salutini et al., 2020). Treatment of DFU remains often challenging and time consuming due to the reported uncomfortable outcomes from both surgical and non-surgical procedures (Smith-Strom et al., 2017). Drug such as amoxicillin, amoxicillin clavulanic acid, ciprofloxacin, dicloxacillin, flucloxacillin, vancomycin and gentamycin are broad spectrum antibiotics used in the treatment of DFI but have side effects that include hypoglycemia, diarrhea, gastrointestinal alteration, weight loss, nephrotoxicity and nausea (Nicolas et al., 2017).

1.3       Justification for the Research

During the last decades, research has aimed at developing effective therapeutic strategies against diabetic foot infections. Medicinal plants have been used as first line therapy for inflammations, burns, ulcers and surgical wound owning to the fact that they contain natural  bioactive  compounds  which  help  hasten  the process  of wound  healing  and regenerate tissue at the wound site. In an attempt to reduce the side effects of drugs and host of other problems associated with DFI, the medicinal properties of Curcuma longa extract and isolated compounds was investigated for their antimicrobial properties against isolates from patients with DFI.

Data generated from this research has contributed to the knowledge of antimicrobial effectiveness on turmeric and validate the traditional claim of the existing data on turmeric as remedy for diabetic foot infection.

1.4       Aim and Objectives

1.4.1    Aim of the study

The aim of the study was to evaluate the antibacterial activity of turmeric (Curcumin longa) extract mediated silver nanoparticles against isolates from patients with diabetic foot infection.

1.4.2    Objectives

The objectives of the study were to:

i.      screen crude extracts of Curcuma longa for the presence of various phytochemical components.

ii.      evaluate antibacterial potentials of crude extracts and extract mediated silver nanoparticles on isolates from patients with diabetic foot infection.

iii.      determine   the   minimum   inhibitory   concentrations   (MIC)   and   minimum bactericidal concentrations (MBC) of extract mediated silver nanoparticles.

iv.      characterize extract mediated silver nanoparticles.

v.        carry out topical toxicological study in rats.

₦8,000.00 – DOWNLOAD FULL PROJECT DOWNLOAD FULL PROJECT Added to cart

---

---

---

---

Related Project Topics :