Cytotoxicity Assay Using Brine Shrimp Lethality Test on CollagenChitosan Wond Dressing Sterilized by Ultraviolet Light Ary Andini1.
Endah Prayekti1.
Devyana Dyah Wulandari1.
Ersalina Nidianti1 Department of Medical Laboratory Technology.
Faculty of Health.
Universitas Nahdlatul Ulama Surabaya.
Surabaya.
Indonesia Correspondence:
Ary Andini.
Jl.
Jemursari No.
Surabaya.
East Java.
Indonesia Zip Code : 60237 Email: aryandini@unusa.
Received: February 14, 2020 Revised: February 17, 2020 Accepted: March 23, 2020 Abstract Collagen gives a moist state on the wound area to accelerate the wound healing process.
Chitosan is a polymer known as non-toxic, antibacterial, antifungal, biodegradable, and biocompatible material.
Combination of collagen and chitosan is expected to be the best biomaterials as a wound dressing for the healing process.
The study aimed to determine the cytotoxicity assay on collagen-chitosan wound dressing sterilized by ultraviolet (UV) Light using Brine Shrimp Lethality Test (BSLT) method.
The test groups were divided into K0.
K1.
K2, and K3 groups.
contained pure chitosan as a control group.
K1 contained collagen 25%-chitosan 75%.
K2 contained collagen 50%chitosan 50%.
K3 contained collagen 75%-Chitosan 25%.
Collagen was extracted from the skin and scalp of snakehead fish (Channa striat.
, then it was mixed with chitosan until collagen-chitosan wound dressing formed.
This study used Brine Shrimp Level Test (BSLT) method with solution concentration: 10, 50, 100, 250, 500, 750 and 1000 ppm.
Based on the results, it showed that K0.
K1.
K2, and K3 group had LC50 > 1000, proved that collagen-chitosan wound dressing was non-toxic material.
The conclusion of the study explain that composite wound dressing based on collagen-chitosan in all groups that was sterilized under UVLight for 15 minutes was not toxic.
Also, it showed LC50 > 1000 based on Brine Shrimp Lethality Test.
Keywords Collagen, chitosan.
BSLT, wound dressing, ultraviolet has main roles in healing the wound and INTRODUCTION There are two kinds of wound dressing, preventing infection on wound area.
Best such as traditional wound dressing including wound dressing with good quality could gauze, plasters, lint, bandages, cotton wool, accelerate the wound healing, prevent the and modern wound dressing including films, wound from infection of bacteria and fungal, and reduce pains .
The particular wound composites .
Wound dressing aplication dressing can promote a newly epithelium Ary Andini, et al.
layer that may not be damaged during the dressing removal around the wound .
The Skin and scales of sneakhead fish were collected from Srikandi fishmonger at Banjar combination of any kind of materials for Asri Village.
Tanggulangin Sub-District.
It is suitable to burn wounds.
Sidoarjo.
East Java Province.
Indonesia.
The surgical wounds, infectious wound and 100 mesh Chitosan powder .
ood and refractory chronic wound .
medical grad.
was made from black tiger Collagen and chitosan are known as MATERIALS AND METHODS shrimp shells and it was obtained from Monodon (Marine Natural Produc.
The including biocompatible and biodegradable cytotoxicity assay was conducted by using .
Therefore, it can be developed into Artemia salina (Golden West Artemia, composite dressing for wound healing.
Supreme Plu.
However, its application is limited due to The collagen was extracted from the skin clinical test before using for human.
Few and scales of sneakhead fish by macerating it steps were needed before clinical test of a in HCl 2% for 48 hours and it was neutralized wound dressing which are toxicity .
, with NaOH 1M.
Chitosan powder was quality .
, in vitro and in vivo evaluation test dissolved in CH3COOH 1% .
Collagen- .
chitosan composite was synthesized by Based on Ariyadi and Dewi .
study, it mixing collagen 25% : chitosan 75% as K1 showed that dry sterilization using ultraviolet collagen 50% : chitosan 50% as K2 (UV) irradiation for 10 and 15 minutes would collagen 75% and chitosan 25% as K3 not be contaminated by Bacillus subtilis and pure chitosan as a control group colony .
Therefore, this research used (K.
composite collagen-chitosan extracted from The cytotoxicity assay was carried out Channa striata collagen that was sterilized with the Brine Shrimp Lethally Test (BSLT) under ultraviolet (UV) light for 15 minutes to by using A.
Each collagen-chitosan determine LC50 using Brine Shrimp Lethally composite of K0.
K1.
K2 and K3 was Test (BSLT).
Previously, the brine shrimp dissolved with DMSO 1 % then was divided was utilized in various bioassay such as into 10 ppm, 50 ppm, 100 ppm, 250 ppm, 500 ppm, 750 ppm and 1000 ppm.
As much as 5 pollutant, anesthetics, dinoflagellate toxins, mL of each concentration was added into mycotoxins, cocarcinogenecity of phorbol petri dish then it was added with 5 ml of esters, and toxicants in marine .
Hereafter, it was incubated for 24 hours .
Ina J Med Lab Sci Tech 2020.
: 21-26
Ary Andini, et al.
% Mortality = Total Larvae Mortality x 100 % Total Larvae RESULTS Brine Shrimp Lethally Test (BSLT) was LC50 assessment was analyzed by Microsoft Probit conducted using Artemia saline larvae to study the toxicity of collagen-chitosan Afterward, the toxicity categories of each This study was a preliminary test sample could be determined based on the before the composite applied in vivo.
The Table 1.
result of probit analysis of larvae mortality Table 1.
LC50 categories .
Categories Very Toxic Toxic Non-Toxic percentage for each group sample can be seen LC50 .
<30 > 1000 in Table 2, 3, 4, and 5.
(Source: Meyer et al, 1.
Table 2.
Percentage of Larvae Mortality in K0 Group Concentration Total Log 10 Mortality .
Larvae 1,00 1,70 2,00 2,40 2,70 3,00 Table 3.
Percentage of Larvae Mortality in K1 Group Concentration Total Log 10 Mortality .
Larvae 1,00 1,70 %Mortality Probit
4,48
3,72
%Mortality Probit
2,00
3,72
2,40
2,70
4,16
3,72
3,00
4,16
Ina J Med Lab Sci Tech 2020.
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Ary Andini, et al.
Table 4.
Percentage of Larvae Mortality in K2 Group Concentration Total Log 10 Mortality .
Larvae 1,00 1,70 2,00 2,40 2,70 3,00 Table 5.
Percentage of Larvae Mortality in K3 Group Concentration Total Log 10 Mortality .
Larvae 1,00 1,70 2,00 2,40 2,70 3,00 Tabel 6.
LC50 Assesment No.
Group
LC50
> 1000
> 1000
> 1000
> 1000
%Mortality Probit
4,48
3,72
4,16
4,16
4,16
%Mortality Probit
4,16
3,72
4,48
3,72
Toxicity Non-Toxic Non-Toxic Non-Toxic Non-Toxic (Source: Meyer et al, 1.
Based on the results, it showed that wound treatment which increase every year.
collagen-chitosan composite sterilized by Chitosan UV light was not toxic.
This is based on the biomaterials for wound dressing because of interpretation of the data, according to Meyer its bioproperties.
Sterilization of materials et al, .
before in vivo assay must be done to prevent microbial and fungal contamination.
The UV
DISCUSSION
Light exposure on materials could mantain The composit dressing is used for wound sterility of materials.
However, not every treatment to reduce the contamination risk.
material suitable for this methods.
Therefore.
The research about wound dressing had cytotoxicity assay has to be conducted to developed rapidly due to the demand for analyse its bioproperties before it used for Brine Shrimp Lethally Test was a Ina J Med Lab Sci Tech 2020.
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Ary Andini, et al.
simple cytotoxicity assay by using A.
promote the rate and quality of diabetic full- to obtain LC50.
If LC50 value is more than thickness wound healing on rats.
Moreover, 1000, it means that the materials tested were collagen from snakehead fish contains not toxic .
glycine, glutamine and arginine.
Glycine has Based on the results of the study, it a role in collagen synthesis around the wound showed that wound dressing for skin wound area, while glutamine has a role as an energy healing that sterilized by UV light at the source during the inflammatory phase and whole group was not toxic because LC50 > proliferation phase of Chitosan was also known as anti- Arginine has a role in immunity system and bacterial and anti-fungal properties .
stimulates endothelial cell function.
Exposure of UV sterilization used was 254 chitosan was known as a biomaterial for drug nm to inactivate the microbe.
Absorption of release on wound area due to its hydrogels UV light on microbe could damage nucleic properties .
Therefore, the combination acids and their constituents that caused of collagen and chitosan can enhance the mutagenetic effect and cell retardation .
bioproperties of collagen-chitosan composite Ultra-Violet (UV) radiation on the material wound healing.
nucleoprotein compounds and occurs the CONCLUSIONS cross-linked of timin molecules that could Collagen-chitosan generate false genetic code, promoting wound dressing in all groups (K0.
K1.
K2 and mutation that will cause the damage and K.
that was sterilized under UV-Light for 15 weaken the vital functions of the organism minutes were not toxic based on Brine until dead .
Shrimp Lethally Test which showed LC50 > Based on the results of Ariyadi and Dewi
study, it showed that the best UV method
as anti-bacterial treatment was obtained in 10
ACKNOWLADGEMENTS
and 15 minutes.
Therefore, this study was The authors gratefully acknowledge the using 15 minute for UV exposure as a financial supports of this research from the sterilization method on the wound dressing.
Institute Collagen could heal chronics wound by Development of Ulama.
Research Community Universitas Nahdlatul endogenous collagen synthesis on wound area .
Based on Lei et al.
research, it CONFLICT OF INTEREST There are no conflicts of interest.
Ina J Med Lab Sci Tech 2020.
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showed that collagen hydrogel dressing could Ary Andini, et al.
REFERENCES