PLANT-DERIVED EV SCAFFOLD AND GTR MEMBRANE FOR ANTIMICROBIAL AND REGENERATIVE EFFECTS

Abstract

The present invention relates to a novel tissue-engineered scaffold modified with plant-derived extracellular vesicles (PDEVs) for periodontal regeneration. PDEVs, isolated from medicinal plant extracts, contain bioactive molecules such as proteins, RNA, and miRNAs, which promote antimicrobial, anti-inflammatory, and regenerative effects. The scaffold is fabricated using a biocompatible polymeric material with optimized porosity for cell infiltration and tissue integration. Functionalization with PDEVs enhances therapeutic properties and ensures biocompatibility. In vitro evaluations confirm the scaffold’s superior antimicrobial activity against Porphyromonas gingivalis and significant anti-inflammatory effects, leading to accelerated tissue healing. Comparative studies demonstrate its enhanced performance over conventional GTR membranes in terms of regeneration and therapeutic potential. This cost-effective and culturally sensitive innovation addresses the limitations of current treatments, offering a sustainable and efficient solution for periodontal disease management and broader applications in regenerative medicine.

Specification

Description:FIELD OF INVENTION
The present invention relates to the regenerative periodontology, specifically to a tissue-engineered scaffold modified with plant-derived extracellular vesicles. The scaffold promotes accelerated periodontal regeneration and provides antimicrobial and anti-inflammatory effects.
BACKGROUND OF THE INVENTION
Periodontitis is a prevalent chronic inflammatory condition affecting the periodontium, leading to gum inflammation, alveolar bone loss, and eventual tooth loss. Current treatment modalities, such as Guided Tissue Regeneration (GTR), utilize scaffolds and bone grafts to regenerate lost periodontal tissues. However, these treatments face significant challenges, including limited effectiveness, technical sensitivity, and risks of secondary inflammation and immune rejection. These limitations necessitate exploring innovative approaches to achieve complete and accelerated periodontal regeneration.
One promising avenue lies in leveraging tissue engineering techniques, particularly the use of extracellular vesicles (EVs). EVs, naturally secreted by cells, are known to carry bioactive molecules such as proteins, RNA, and miRNAs, which play a crucial role in tissue repair and immunomodulation. Although animal- or human-derived EVs have shown potential, their cost, ethical concerns, and limited availability restrict their widespread use. Plant-derived extracellular vesicles (PDEVs), on the other hand, offer a sustainable and biocompatible alternative with similar therapeutic benefits. However, their application in periodontal regeneration remains largely unexplored.
This invention addresses the existing challenges by developing a cost-effective, biocompatible scaffold modified with PDEVs to enhance periodontal tissue regeneration. The scaffold is designed to not only promote accelerated healing but also provide antimicrobial and anti-inflammatory effects, crucial for preventing disease progression. By leveraging the bioactive properties of PDEVs and addressing cultural apprehensions related to animal-derived products, this invention offers a practical, innovative solution to the limitations of current periodontal treatment strategies.
OBJECTS OF THE INVENTION
Some of the objects of the present disclosure, which at least one embodiment herein satisfies, are as follows.
It is an object of the present disclosure to ameliorate one or more problems of the prior art or to at least provide a useful alternative
An object of the present disclosure is to isolate and purify extracellular vesicles (EVs) from medicinal plant extracts.
Another object of the present disclosure is to characterize plant-derived EVs for their protein, RNA, and therapeutic properties.
Still another object of the present disclosure is to fabricate a biocompatible scaffold with optimized porosity for tissue regeneration.
Another object of the present disclosure is to functionalize the scaffold surface with plant-derived EVs for enhanced bioactivity.
Still another object of the present disclosure is to Provides enhanced antimicrobial activity against periodontal pathogens like Porphyromonas gingivalis.
Still another object of the present disclosure is to demonstrates significant anti-inflammatory effects, aiding in tissue healing and repair.
Yet another object of the present disclosure is to accelerates periodontal regeneration through the bioactive properties of plant-derived EVs.
Yet another object of the present disclosure is to offers a cost-effective alternative to traditional GTR membranes.
Yet another object of the present disclosure is to eliminates risks of immune rejection associated with animal-derived products.
Yet another object of the present disclosure is to utilizes a biocompatible scaffold design with optimal porosity for cell infiltration and integration.
Yet another object of the present disclosure is to combines therapeutic and regenerative functions, reducing the need for additional treatments.

Other objects and advantages of the present disclosure will be more apparent from the following description, which is not intended to limit the scope of the present disclosure.

SUMMARY OF THE INVENTION
The following presents a simplified summary of the invention in order to provide a basic understanding of some aspects of the invention. This summary is not an extensive overview of the present invention. It is not intended to identify the key/critical elements of the invention or to delineate the scope of the invention. Its sole purpose is to present some concept of the invention in a simplified form as a prelude to a more detailed description of the invention presented later.

The present invention is generally relates to a biocompatible scaffold modified with plant-derived extracellular vesicles (PDEVs) for accelerated periodontal regeneration and disease prevention. The scaffold integrates bioactive molecules such as proteins, RNA, and miRNAs from PDEVs to enhance therapeutic potential.
An embodiment of the present invention plant extracts rich in medicinal properties are used as the source for PDEVs, which are isolated and purified through ultracentrifugation and characterized using advanced techniques like DLS and TEM to ensure efficacy and bioactivity.
Another embodiment of the invention the scaffold is fabricated using a polymeric material with optimized porosity, designed to support cell infiltration, nutrient exchange, and effective bone tissue integration, addressing the limitations of current GTR membranes.
Yet another embodiment of the invention is functionalization of the scaffold with PDEVs is achieved to ensure a uniform and stable distribution of bioactive compounds. This modification enhances the scaffold's regenerative, antimicrobial, and anti-inflammatory properties.
Yet another embodiment of the invention is in vitro studies confirm the scaffold's antimicrobial efficacy against Porphyromonas gingivalis, a key pathogen in periodontitis, demonstrating its ability to reduce bacterial colonization effectively.
Yet another embodiment of the invention is the anti-inflammatory activity of the scaffold is validated through cytokine profiling and wound-healing assays, showing reduced inflammatory markers and accelerated tissue repair.
DETAILED DESCRIPTION OF THE INVENTION
The following description is of exemplary embodiments only and is not intended to limit the scope, applicability or configuration of the invention in any way. Rather, the following description provides a convenient illustration for implementing exemplary embodiments of the invention. Various changes to the described embodiments may be made in the function and arrangement of the elements described without departing from the scope of the invention.
The present invention relates to a novel tissue-engineered scaffold modified with plant-derived extracellular vesicles (PDEVs) to enhance periodontal regeneration. The scaffold is fabricated using a biocompatible polymeric base material with optimized porosity to support cell infiltration and tissue integration. PDEVs, rich in bioactive molecules such as proteins, RNA, and miRNAs, are isolated from medicinal plant extracts and integrated onto the scaffold surface through functionalization techniques. This modification endows the scaffold with enhanced regenerative properties and antimicrobial and anti-inflammatory effects. The invention leverages the immunomodulatory and therapeutic potential of PDEVs to accelerate tissue repair and inhibit Porphyromonas gingivalis, a primary pathogen in periodontitis. The scaffold provides a cost-effective and biocompatible solution that overcomes the limitations of traditional GTR membranes, such as immune rejection and limited applicability, while catering to the cultural and ethical sensitivities of diverse patient populations.
EXAMPLE 1: Preparation of Plant-Derived Extracellular Vesicles
Plant-derived extracellular vesicles (PDEVs) are prepared by selecting medicinal plant extracts rich in bioactive compounds based on their therapeutic potential. The EVs are isolated from these extracts using ultracentrifugation, followed by purification and characterization techniques such as dynamic light scattering (DLS) and transmission electron microscopy (TEM). Further analysis of the EVs' cargo, including proteins and RNA, is performed to confirm their suitability for promoting tissue regeneration and addressing inflammatory conditions.
EXAMPLE 2: Fabrication of the Scaffold
A biocompatible scaffold is fabricated using a polymeric base material tailored for tissue engineering applications. The scaffold is designed with optimal porosity to ensure efficient cell infiltration, nutrient exchange, and bone tissue integration. Its structural properties are engineered to support the sustained release of therapeutic agents and facilitate periodontal tissue regeneration.
EXAMPLE 3; Surface Modification of the Scaffold
The surface of the fabricated scaffold is modified with PDEVs to enhance its regenerative and therapeutic properties. This functionalization is achieved through immobilization techniques that ensure a uniform distribution of PDEVs across the scaffold surface. The modified scaffold is then characterized to confirm EV attachment, structural stability, biocompatibility, and bioactivity, ensuring its effectiveness in clinical applications.
EXAMPLE 4: In Vitro Evaluation
The PDEV-modified scaffold is subjected to in vitro testing to evaluate its therapeutic efficacy. Antimicrobial activity is assessed against Porphyromonas gingivalis, a key pathogen involved in periodontitis, while anti-inflammatory effects are studied through cytokine profiling and wound-healing assays. These evaluations determine the scaffold's ability to combat infection and promote tissue repair.

The PDEV-modified scaffold demonstrated enhanced antimicrobial activity against Porphyromonas gingivalis compared to commercially available GTR membranes, effectively reducing bacterial growth. It also exhibited significant anti-inflammatory effects, as shown by reduced pro-inflammatory cytokine levels and improved wound healing in in vitro assays.
Comparison with Commercial GTR Membranes
The efficacy of the PDEV-modified scaffold is compared to commercially available guided tissue regeneration (GTR) membranes. The comparison focuses on antimicrobial activity, anti-inflammatory effects, and regenerative performance, demonstrating the advantages of the PDEV-modified scaffold in terms of enhanced biocompatibility, effectiveness, and therapeutic potential for periodontal applications.

While considerable emphasis has been placed herein on the specific features of the preferred embodiment, it will be appreciated that many additional features can be added and that many changes can be made in the preferred embodiment without departing from the principles of the disclosure. These and other changes in the preferred embodiment of the disclosure will be apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.
, Claims:1. A tissue-engineered scaffold comprising a polymeric base material modified with extracellular vesicles derived from plant extracts, wherein the scaffold facilitates periodontal regeneration and provides antimicrobial and anti-inflammatory effects.
2. The scaffold as claimed in claim 1, wherein the extracellular vesicles are characterized by their lipid bilayer structure and cargo of proteins, RNA, and miRNAs.
3. A method of fabricating the scaffold as claimed in claim 1, comprising:
a) isolating and purifying extracellular vesicles from plant extracts.
b) fabricating a porous scaffold using a polymeric material.
c) functionalizing the scaffold surface with extracellular vesicles.

Dated this 23 November 2024

Dr. Amrish Chandra
Agent of the applicant
IN/PA No: 2959

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