DEVELOPMENT OF A HYBRID BIOINK: INCORPORATION OF COPPER OXIDE NANOPARTICLES AND CARBON QUANTUM DOTS

Abstract

Development of a Hybrid Bioink: incorporation of Copper Oxide nanoparticles and Carbon Quantum Dots inside a Chitosan matrix for Biomedical purposes ABSTRACT The bioink claimed herein, may be employed for a wide spectrum of industrial applications, including tissue engineering, wound healing, drug delivery and biomedical imaging, while providing a greater level of functionality over standard bioinks. Moreover, this bioink can be readily scaled for large-scale production. The hybrid bioink can be engineered to provide multiple functions, while reducing the need for additional components and thus simplifying fabrication.

Specification

FORM - 2
THE PATENTS ACT, 1970 (39 OF 1970)
&
THE PATENTS RULES,
2003 COMPLETE SPECIFICATION
(see Section 10 & rule 13)

1. TITLE OF THE INVENTION : Development of a Hybrid Bioink: incorporation of Copper Oxide
nanoparticles and Carbon Quantum Dots inside a Chitosan matrix for Biomedical purposes
2. APPLICANT:
S.No NAME NATIONALITY ADDRESS
1 Saveetha Institute of Medical and
Technical Sciences
INDIAN Saveetha Nagar, Thandalam,
Chennai - 602 105,Tamil Nadu,
India

3. PREAMBLE TO THE DESCRIPTION:
The following specification describes the invention and how it is to be performed,
4. COMPLETE SPECIFICATION
The following specification particularly describes the invention and the manner in which it is tobe
performed. ....................... Separate sheet is attached......................................
5. DESCRIPTION
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6. CLAIMS
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DATE:
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NAME
1. ABSTRACT OF THE INVENTION
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Bank : INDUSIND BANK
Branch : NUNGAMBAKKAM
( SAVEETHA UNIVERSITY )
DD No : 402553
DD Date: 14/11/2024
DD Amount: 8,900/-

Development of a Hybrid Bioink: incorporation of Copper Oxide nanoparticles and Carbon
Quantum Dots inside a Chitosan matrix for Biomedical purposes
PREAMBLE TO THE DESCRPTION

Field of the Invention:
The main aim of this invention is producing a bioink for application in biomedical fields that
includes 3D bioprinting, tissue scaffolds, drug delivery and wound healing. This hybrid bioink
consists of chitosan as the primary matrix, incorporated with copper oxide nanoparticles and carbon quantum dots (CQDs) which improves the bioink's antibacterial, mechanical and imaging
characteristics Incorporating bioinks into bioprinting and tissue engineering will enable the
fabrication of tissue, skin, and organ structures containing cells, biomolecules, and biomaterials for regenerative medicine and cellular therapies in a scalable manner.

Background of the Invention:
Bioinks have come a long way from its inception, and tissue engineering, regenerative medicine has advanced ever since [l]. Chitosan is a natural biopolymer obtained by deacetylation of chitin and has good skin compatibility, biodegradability as well as antimicrobial properties However, typical bioinks based only on chitosan frequently have limits in mechanical strength, stability, and
multifunctionality [2].

Nowadays the development of bioink can be enhanced by addition nanoparticles with particular properties, which among them manly are exploitation nanoparticles originating from
current advances in nanotechnology. Copper oxide nanoparticles (CuO NPs) have significant strong
antibacterial activity, and therefore these can play a potential role in reducing infection in tissue engineering [3]. On the other hand, carbon quantum dots (CQDs) have shown particular fluorescence properties that can be employed to label and trace the migration or development of cells in a variety of contexts, from microenvironments such as scaffolds in vitro to studies grounded on tissue engineering approaches in vivo [4].

While studies reported on chitosan and nanoparticles separately as materials, there has been very few research that incorporates an approach that seeks to chemically incorporate the particles into chitosan to develop a functional bioink. The incorporation of the hybrid bioink presents a foundational solution to fill voids in creating a multifunctional bioink for biomedical applications, including tissue engineering, wound healing, drug delivery, bioimaging, and other biotechnological opportunities [5].

Summary of the Invention:
This bioink presents multiple advantages due to the layers of components added: chitosan,
copper oxide nanoparticles, and carbon quantum dots. In combination, copper oxide nanoparticles
significantly improve antibacterial properties within the chitosan matrix, while CQDs provide a
fluorescent means to facilitate efficient real-time bioimaging. Synthesizing a hybrid bioink provides a multifunctional material that serves as a better bioink for 3D bioprinting, along with desirable mechanical properties, biocompatibility and easily modifiable based on applications.

Development of a Hybrid Bioink: incorporation of Copper Oxide nanoparticles and Carbon
Quantum Dots inside a Chitosan matrix for Biomedical purposes
COMPLETE SPECIFICATION

Specifications
1. Preparation of Copper Oxide Nanoparticles
Copper oxide nanoparticles are produced using a chemical precipitation method. More
specifically, copper salts (CUSO4) and base (NaOH) are combined in an aqueous mixture. The copper
hydroxide produced is heated to produce copper oxide nanoparticles. The size of the produced nanoparticles may be further modified with different reaction conditions, including the concentration of reactants, the temperature of the reaction, and the time [8].

2. Synthesis of Carbon Quantum Dots
Carbon Quantum Dots (CQDs) arc using hydrothermal method. A carbon source, such as citric acid or a simple sugar, is sealed and heated in a high-pressure autoclave mode (-150-200 °C) to synthesize carbon in the aqueous structure. CQDs are purified by dialysis treatment to remove
organic residual. The CQDs were subsequently functionalized with of surface groups, of amine or
carboxyl groups to invest in solid dispersion in chitosan increase biocompatibility of the CQDs [9].

3. Preparation of Chitosan Matrix:
Chitosan solution was prepared by dissolving chitosan in dilute acetic acid solution (1-2%
v/v), this produces a transparent solution. The pH of the solution is adjusted to 6-7 range to achieve optimal conditions for crosslinking. For enhancement of the matrix and mechanical properties prior to 3D printing, crosslinkers such as sodium tripolyphosphate (TPP) can be added. This chitosan matrix acts as biocompatible base for bioink which is easy to fabricate, and this also supports cell growth. Carbon quantum dots and copper oxide nanoparticles were incorporated in this chitosan solution [10].

Development of a Hybrid Bioink: incorporation of Copper Oxide nanoparticles and Carbon Quantum Dots inside a Chitosan matrix for Biomedical purposes

DESCRIPTION
Bioink Formulation:
Copper oxide nanoparticles and carbon quantum dots were dispersed throughout the chitosan solution. Then it was stirred at room temperature for homogenous dispersion [l I]. This helps in
preventing agglomeration of nanoparticles and maintaining bioink properties.
• The bioink composition contain 0.1 -1 % copper oxide and 0.05-0.5% carbon quantum dots.
• Chitosan concentration: 1-3% w/v, based on desired viscosity and mechanical strength.
The prepared bioink has appropriate viscosity for better extrusion-based 3D bioprinting, while
maintaining the shear characteristics making it as ideal bioink for tissue scaffolds bioprinting.

5. 3D Bioprinting and Applications:
The prepared bioink is placed into a 3D bioprinter and extruded into specific shapes for tissue
engineering purposes. The bioink has exceptional printability, retaining form accuracy during and
after printing.
• Tissue Scaffolds: Bioink may be printed on porous scaffolds for cell culture and tissue
regeneration. Copper oxide nanoparticles reduce bacterial contamination, whereas carbon quantum dots provide real-time imaging.
• Wound Healing: These bioinks are directly placed on wounds for wound acceleration and monitoring the wounding healing process via fluorescence.
• Drug Delivery Systems: Bioinks may be produced to encapsulate therapeutic chemicals allowing for both therapy and imaging [12,13]

6. Optimization and Testing:
1. Rheological testing is done to access the viscosity density and shear-stress characteristics of
bioink, which is mainly responsible for optimal printing. Viscosity is optimized between
1,000 and 5,000 Pa.s for smooth extrusion.
2. Tensile strength testing is used to check ductility of material which is suitable for tissue
engineering.
3. Antibacterial Testing: Copper oxide nanoparticle shows reduced microbial growth, thus
preparing the scaffold with anti-microbial property.
4. Cell Compatibility: In vitro experiments with fibroblasts and stem cells show the bioink is
biocompatible and promotes cell growth [14].

Development of a Hybrid Bioink: incorporation of Copper Oxide nanoparticles and Carbon
Quantum Dots inside a Chitosan matrix for Biomedical purposes

CLAIM
We Claim

1. Claim l:
This bioink contains chitosan, copper oxide nanoparticles, and carbon quantum dots.

2. Claim 2:
As in the claim l, the synthesized copper nanoparticle have an average size of 10-50 nm
which exhibits antibacterial activity

3. Claim 3:
The bioink of claim 1, comprising the carbon quantum dots emit fluorescence, allowing for
real-time bioimaging during biomedical applications.

4. Claim 4:
A process for producing a hybrid bioink includes:
❖ Synthesizing copper oxide nanoparticles via chemical precipitation.
❖ Carbon quantum dots synthesized with hydrothermal method.
❖ Chitosan is dissolved in acetic acid to produce a matrix.
❖ Incorporating copper oxide nanoparticles and carbon quantum dots to the chitosan matrix.

5. Claim 5:
The bioink described in the claim 1 is used for drug delivery, wound healing, tissues and scaffold engineering applications

6. Claim 6:
A process of creating the bioink specified in claim 1 includes testing for rheological
characteristics, mechanical strength, and antibacterial activity.

7. Claim 7:
The bioink of claim 1 has a composition tuned-for shear-thinning behavior, enabling for regulated
extrusion during 3D bioprinting.

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