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Abstract

Diabetes mellitus is a chronic metabolic disorder affecting millions worldwide, with projections indicating a rise in prevalence by 2045. To address this global health issue, recent research has turned to nanotechnology, particularly the use of silver nanoparticles (AgNPs) synthesized via green methods. This article highlights the biosynthesis of AgNPs using the leaf extract of Costus igneus, commonly known as the insulin plant, and evaluates their anti-diabetic potential both in vitro and in vivo.

Keywords

Diabetic, Silver nanoparticles, Green Synthesis, streptozosine, Costus ingnius etc.

Introduction

Diabetes mellitus is a chronic metabolic disorder characterized by high levels of blood glucose, caused by the inability of the body to produce insulin or by the ineffective use of the insulin produced. According to International Diabetes Federation (IDF), globally, as of 2019, approximately 463 million people were living with diabetes, which rise to 700 million by 2045.In recent years, application of nanomaterial in antidiabetic studies has drawn attention due to their extraordinary features such as ultra-small size, ability to transport the therapeutic agent through the cell membrane at the target site and bio-adaptability. Green/biological synthesis ( using plant, bacteria, algae and fungi) of AgNPs is preferred chemical and physical methods, since it is an environmentally friendly and cost-effective method without the use of high temperature, pressure, toxic chemicals. Plant (Phyto) mediated fabrication of AgNPs has gained popularity among the other green approaches since it is easily available in large quantities, contains secondary metabolites, and negligible cross-contamination between the plant extract. Hence in the present study, Costus igneus was selected for fabrication of AgNPs.

Costus igneous (insulin plant) is used as an herbal cure for diabetes in India. Very few studies have been conducted on the biosynthesis of silver and zinc oxide nanoparticles using Costus igneus leaves extract and evaluation of their antioxidant, antimicrobial and anticancer activities. However, the anti-diabetic potential of AgNPs fabricated with Costus igneus leaves extract has so

far not been reported.

       
            Image of Costus igneous (Insulin) Plant.png
       

 Image of Costus igneous (Insulin) Plant.

Aqueous Extraction and synthesis of Costus igneous AgNPs.

       
            Schematic illustration of the green synthesis of Costus igneus AgNPs.png
       

 Figure No.5 Schematic illustration of the green synthesis of Costus igneus AgNPs

Results: -

Phytochemical analysis:

The results of qualitative screening of phytochemical constituents present in the aqueous extract of Costus igneus are shown phytochemical profile of aqueous leaf extract of Costus igneus revealed the presence of carbohydrates, proteins, steroids, glycosides, flavonoids, alkaloids, and tannins which may be responsible for the efficient capping and stabilization of nanoparticle.

Visual observation:

The formation of         CI-AgNPs       was confirmed by observing the         colour  change  of  the  colloidal solution from         light yellow to dark brown.

       
            UV-Vis spectral analysis.png
       

UV-Vis spectral analysis:

The UV-Vis spectrum of colloidal solution of CI-AgNPs showed an absorption peak at 436nm.

           
            Fourier transform infrared analysis.png
       

 Fourier transform infrared analysis (FTIR):

       
            Particle size and Zeta potential.jpg
       

  A. Fourier transform infrared analysis of AECI Extract. B. Fourier transform infrared spectra of CI-AgNPs

Particle size and Zeta potential:

The biosynthesized silver nanoparticles have a particle size of 86.50nm and Zeta potential of -25.8 mV.

       
            pic-2.png
       

Transmission electron microscopy (TEM):

The silver nanoparticles were found to be roughly textured and grossly spherical in shape and the average particle size was found to be 86.50nm

       
            Energy dispersive x-ray spectroscopy.png
       

Energy dispersive x-ray spectroscopy (EDX):

The EDX spectrum of AECI-AgNPs shows a signal peak at 3 keV which is typically for elemental silver.

       
            Selected area electron diffraction.png
       

Selected area electron diffraction (SAED):

The SAED pattern was used to determine the crystalline structure of silver nanoparticles synthesized using AECI as reducing agent the image revealed the ring patterns appeared with light spots on the dark field.

       
            X-ray diffraction analysis.png
       

X-ray diffraction analysis (XRD):

The XRD pattern of AECI-AgNPs showed the Bragg’s diffraction peaks at 38.08°, 46.29°, and 77.08° respectively corresponding to (111), (200), and (300) planes of the face centered cubic lattice. The XRD analysis data conformed the SAED results.

       
            pic-3.png
       

. In Vitro Anti-Diabetic Activity*:

?-Amylase Inhibition Assay*: The AgNPs exhibited significant inhibition of ?-amylase, an enzyme involved in carbohydrate digestion, with an IC50 value of 105.10 µg/ml, making them potentially effective for diabetes management.

Glucose Uptake Assay: The glucose uptake by yeast cells was higher in the presence of AgNPs (78.04%) compared to the aqueous extract alone (49.78%), demonstrating their potential in regulating blood glucose levels.

In-vivo antidiabetic activity was assessed in STZ induced rats:

Effect of AE-CI, CI-AgNPs on lipid profiles (TC, TG, LDL and HDL)


Groups

Treatments

TC

mg/dl

TG

mg/dl

HDL

mg/dl

LDL

mg/dl

I

Normal control

68.33 ± 2.76

81.1 ± 4.15

54.04 ± 3.67

33.7 ± 2.19

II

Diabetic control

102.1 ± 5.03

161.4 ± 4.87

39.11 ± 1.83

115.5 ± 3.97

III

Diabetic + Glimepiride (15 µg/kg b.w)

75.19 ± 5.80

90.64 ± 4.41

51.14 ± 2.13

44.79 ± 2.47

IV

Diabetic + AE-CI (100 mg/kg b.w)

89.42 ± 4.29

107.6 ± 3.01

43.77 ± 1.34

65.56 ± 3.13

V

Diabetic + CI-AgNPs (10 mg/kg b.w)

81.66 ± 5.94

96.23 ± 4.13

49.7 ± 2.50

57.66 ± 2.81


The in-vivo antidiabetic activity in STZ induced diabetic rats proved that AECI and CI-AgNPs improved the diabetic dyslipidemia.

Effect of AE-CI, CI-AgNPs on body weight and blood glucose in streptozotocin induced diabetic rats.


Groups

Body weight

Blood glucose

1day

7thday

14th day

21thday

1day

7th day

14thday

21th day

Normal control

185.87±

45.42

190.91±

91

194.81±

30.50

197.72±

42.77

107.33±

3.21

103.94±

2.68

100.36±

2.60

109.49±

1.72

Diabetic

197.17±

178.07±

163.46±

154.03±

374.41±

333.68±

310.23±

286.39±

control

38.21

30.06

17.90

10.58

11.03

48.23

24.91

24.25

 

 

 

 

 

 

 

.

 

Std.

194.40±

187.48±

182.96±

178.70±

356.14±

244.72±

195.64±

163.30±

Glimepiride

39.93

31.21

8.81

13.25

8.32

29.08

12.00

8.83

Diabetic+

206.03±

200.05±

193.85±

186.19±

366.24±

265.37±

237.24±

204.38±

AE-CI(100

49.64

26.38

76.00

56.71

8.53

40.72

54.66

25.37

mg/kg b.w)

 

 

 

 

 

 

 

 

Diabetic+

184.41±

181.53±

177.03±

173.33±

360.84±

248.87±

225.94±

177.49±

C.I-AgNPs

13.34

16.47

33.15

20.14

13.98

26

49.41

32.80

(10mg/kg

 

 

 

 

 

 

 

 

b.w)

 

 

 

 

 

 

 

 


Histopathology Examination:-

Figure No.20. The liver section of histopathology (A) Group I – Normal control (B) Group II- Diabetic control group (C) Group III- Glimepiride treated group (D) Group IV- AE-CI treated group (E) Group V-. CI-AgNPs treated group. Arrows indicate fatty liver central venous congestion along with chronic cholangitis.

       
            Figure No.20. The liver section of histopathology.png
       

The kidney section of histopathology:a(A) Group I – Normal control (B) Group II- Diabetic control group (C) Group III- Glimepiride treated group (D) Group IV- AE-CI treated group (E) Group V-. CI-AgNPs treated group. Arrows indicate few glomeruli show mild mesangial widening, thickening of basement membrane and mild interstitial nephritis.

       
            pic-4.png
       

The pancreas section of histopathology (A) Group I – Normal control

(B) Group II- Diabetic control group (C) Group III- Glimepiride treated group (D) Group IV- AE-CI treated group (E) Group V-. CI-AgNPs treated group. Arrows indicate reduction in number of islets of Langerhans with    islets   showing in distinct                    borders.

       
            pic-5.png
       

REFERENCES

  1. https://www.idf.org/aboutdiabetes/what-is-diabetes/facts-figures.html (Accessed on 20-02 2020)
  2. Jiang J, Pi J, Cai J. The advancing of zinc oxide nanoparticles for biomedical applications. Bioinorganic chemistry and applications. 2018;2018.
  3. Das G, Patra JK, Debnath T, Ansari A, Shin HS. Investigation of antioxidant, antibacterial, antidiabetic, and cytotoxicity potential of silver nanoparticles synthesized using the outer peel extract of Ananas comosus (L.). PloS one. 2019;14(8).
  4. Rafique M, Sadaf I, Rafique MS, Tahir MB. A review on green synthesis of silver nanoparticles and their applications. Artificial cells, nanomedicine, and biotechnology. 2017 ;45(7):1272-91.
  5. Khan T, Khan MA, Nadhman A. Synthesis in plants and plant extracts of silver nanoparticles with potent antimicrobial properties: current status and future prospects. Applied microbiology and biotechnology. 2015; 99(23): 9923-34.
  6. Hegde PK, Rao HA, Rao PN. A review on Insulin plant (Costus igneus Nak). Pharmacognosy reviews. 2014;8(15):67.
  7. Shetty AJ, Choudhury D, Rejeesh VN, Kuruvilla M, Kotian S. Effect of the insulin plant (Costus igneus) leaves on dexamethasone-induced hyperglycemia. International journal of Ayurveda research. 2010;1(2):100.
  8. Aruna A, Karthikeyan V, Bose P. Synthesis and characterization of silver nanoparticles of insulin plant (costus pictus d. don) leaves. Asian journal of biomedical and pharmaceutical sciences. 2014; 4(34).
  9.  Suresh J, Pradheesh G, Alexramani V, Sundrarajan M, Hong SI. Green synthesis and characterization of zinc oxide nanoparticle using insulin plant (Costus pictus D. Don) and investigation of its antimicrobial as well as anticancer activities. Advances in Natural Sciences: Nanoscience and Nanotechnology. 2018; 9(1): 015008.
  10. Prabhu S, Vinodhini S, Elanchezhiyan C, Rajeswari D. Evaluation of antidiabetic activity of biologically synthesized silver nanoparticles using Pouteria sapota in streptozotocin?induced diabetic rats: Journal of diabetes. 2018; 10(1): 28-42.
  11. Shanker K, Mohan GK, Hussain MA, Jayarambabu N, Pravallika PL. Green biosynthesis, characterization, in vitro antidiabetic activity, and investigational acute toxicity studies of some herbal-mediated silver nanoparticles on animal models. Pharmacognosy magazine. 2017 ;13(49):188.
  12. Balan K, Qing W, Wang Y, Liu X, Palvannan T, Wang Y, Ma F, Zhang Y. Antidiabetic activity of silver nanoparticles from green synthesis using Lonicera japonica leaf extract. Rsc Advances. 2016;6(46):40162-8.
  13. Govindappa M, Hemashekhar B, Arthikala MK, Rai VR, Ramachandra YL. Characterization, antibacterial, antioxidant, antidiabetic, anti-inflammatory and antityrosinase activity of green synthesized silver nanoparticles using Calophyllum tomentosum leaves extract. Results in Physics. 2018; 9: 400-8.
  14. Saratale RG, Shin HS, Kumar G, Benelli G, Kim DS, Saratale GD. Exploiting antidiabetic activity of silver nanoparticles synthesized using Punica granatum leaves and anticancer potential against human liver cancer cells (HepG2). Artificial cells, nanomedicine, and biotechnology. 2018; 46(1): 211-22.
  15. Kitture R, Chordiya K, Gaware S, Ghosh S, More PA, Kulkarni P, Chopade BA, Kale SN. ZnO nanoparticles-red sandalwood conjugate: a promising anti-diabetic agent. Journal of nanoscience and nanotechnology. 2015;15(6):4046-51.
  16. Rajaram K, Aiswarya DC, Sureshkumar P. Green synthesis of silver nanoparticle using Tephrosia tinctoria and its antidiabetic activity. Materials Letters. 2015;138:251-4.

Reference

  1. https://www.idf.org/aboutdiabetes/what-is-diabetes/facts-figures.html (Accessed on 20-02 2020)
  2. Jiang J, Pi J, Cai J. The advancing of zinc oxide nanoparticles for biomedical applications. Bioinorganic chemistry and applications. 2018;2018.
  3. Das G, Patra JK, Debnath T, Ansari A, Shin HS. Investigation of antioxidant, antibacterial, antidiabetic, and cytotoxicity potential of silver nanoparticles synthesized using the outer peel extract of Ananas comosus (L.). PloS one. 2019;14(8).
  4. Rafique M, Sadaf I, Rafique MS, Tahir MB. A review on green synthesis of silver nanoparticles and their applications. Artificial cells, nanomedicine, and biotechnology. 2017 ;45(7):1272-91.
  5. Khan T, Khan MA, Nadhman A. Synthesis in plants and plant extracts of silver nanoparticles with potent antimicrobial properties: current status and future prospects. Applied microbiology and biotechnology. 2015; 99(23): 9923-34.
  6. Hegde PK, Rao HA, Rao PN. A review on Insulin plant (Costus igneus Nak). Pharmacognosy reviews. 2014;8(15):67.
  7. Shetty AJ, Choudhury D, Rejeesh VN, Kuruvilla M, Kotian S. Effect of the insulin plant (Costus igneus) leaves on dexamethasone-induced hyperglycemia. International journal of Ayurveda research. 2010;1(2):100.
  8. Aruna A, Karthikeyan V, Bose P. Synthesis and characterization of silver nanoparticles of insulin plant (costus pictus d. don) leaves. Asian journal of biomedical and pharmaceutical sciences. 2014; 4(34).
  9.  Suresh J, Pradheesh G, Alexramani V, Sundrarajan M, Hong SI. Green synthesis and characterization of zinc oxide nanoparticle using insulin plant (Costus pictus D. Don) and investigation of its antimicrobial as well as anticancer activities. Advances in Natural Sciences: Nanoscience and Nanotechnology. 2018; 9(1): 015008.
  10. Prabhu S, Vinodhini S, Elanchezhiyan C, Rajeswari D. Evaluation of antidiabetic activity of biologically synthesized silver nanoparticles using Pouteria sapota in streptozotocin?induced diabetic rats: Journal of diabetes. 2018; 10(1): 28-42.
  11. Shanker K, Mohan GK, Hussain MA, Jayarambabu N, Pravallika PL. Green biosynthesis, characterization, in vitro antidiabetic activity, and investigational acute toxicity studies of some herbal-mediated silver nanoparticles on animal models. Pharmacognosy magazine. 2017 ;13(49):188.
  12. Balan K, Qing W, Wang Y, Liu X, Palvannan T, Wang Y, Ma F, Zhang Y. Antidiabetic activity of silver nanoparticles from green synthesis using Lonicera japonica leaf extract. Rsc Advances. 2016;6(46):40162-8.
  13. Govindappa M, Hemashekhar B, Arthikala MK, Rai VR, Ramachandra YL. Characterization, antibacterial, antioxidant, antidiabetic, anti-inflammatory and antityrosinase activity of green synthesized silver nanoparticles using Calophyllum tomentosum leaves extract. Results in Physics. 2018; 9: 400-8.
  14. Saratale RG, Shin HS, Kumar G, Benelli G, Kim DS, Saratale GD. Exploiting antidiabetic activity of silver nanoparticles synthesized using Punica granatum leaves and anticancer potential against human liver cancer cells (HepG2). Artificial cells, nanomedicine, and biotechnology. 2018; 46(1): 211-22.
  15. Kitture R, Chordiya K, Gaware S, Ghosh S, More PA, Kulkarni P, Chopade BA, Kale SN. ZnO nanoparticles-red sandalwood conjugate: a promising anti-diabetic agent. Journal of nanoscience and nanotechnology. 2015;15(6):4046-51.
  16. Rajaram K, Aiswarya DC, Sureshkumar P. Green synthesis of silver nanoparticle using Tephrosia tinctoria and its antidiabetic activity. Materials Letters. 2015;138:251-4.

Photo
Shivani Kumari
Corresponding author

Shridhar University, Pilani

Photo
Akshay Sharma
Co-author

Haridwar University, Roorkee, Uttarakhand

Photo
Dheeraj Kumar Vishwakarma
Co-author

Haridwar University, Roorkee, Uttarakhand

Dheeraj Kumar Vishwakarma*, Akshay Sharma, Shivani Kumari, Phyto-Fabrication, Characterization and Anti-Diabetic Activity of Silver Nanoparticles Using Costus Igneus (Insulin Plant) Leaf Extract, Int. J. of Pharm. Sci., 2024, Vol 2, Issue 12, 1324-1330. https://doi.org/10.5281/zenodo.143781568

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