Exploring the Integration of Artepillin C, a Bioactive Constituent of Propolis, in Dental Materials: A Review

Introduction

Artepillin C (ARC) is a major bioactive compound derived from Propolis, a resinous substance collected by honeybees from various botanical sources. It possesses a broad spectrum of pharmacological benefits, including antioxidant, antimicrobial, anti-inflammatory, anti-diabetic, neuroprotective, gastroprotective, immunomodulatory, and anti-cancer properties.¹ Most studies have focused on its potential in combating oxidative stress, inflammation, diabetes, and cancer using a combination of in vitro and in vivo methodologies.^2,3 Due to its multiple health benefits—such as antimicrobial activity, antioxidant effects, anti-inflammatory action, inhibition of biofilm formation, and promotion of tissue regeneration, ARC has also attracted growing interest in the field of dental materials.^4,5

Artepillin C (ARC) is a prenylated derivative of p-coumaric acid is found predominantly in Brazilian green propolis (BGP).⁶ compound is mainly derived from Baccharis dracunculiforia plants in Brazil, but has also been identified in Brazilian brown propolis, and various other plant species.⁶

Research has identified p21-activated kinase 1 (PAK1) as a key molecular target of ARC. PAK1 has emerged as a potential target for ARC due its implications in various diseases, including inflammation, diabetes, and cancer ⁷. Maruta and He have proposed that inhibiting PAK1 could be a promising antiviral treatment for COVID-19, presenting a novel therapeutic opportunity for Artepillin C.^7-9 The worldwide recognition of ARC has increased the market value of Propolis, with the concentration of ARC being a benchmark for propolis quality.¹⁰ This review aims to summarize the chemical characteristics, pharmacokinetics profile, bioavailability, oral health benefits, and molecular targets of ARC, particularly in the context of dental materials, while outlining future directions for research. 

The Chemical Characteristics of ARC

ARC is a type of phenolic acid with a simpler name (E)-artepillin C. It weighs about 300.4 g/mol and has two prenyl groups attached to its phenyl ring.⁶ There are two forms of ARC, E (Figure 1) and Z (Figure 2), but the E form is more common and found in Brazilian propolis. It can be sensitive to light and heat, and exposure to sunlight for 5 days turns it into the Z form. If exposed for more than 5 days, it keeps breaking down. Heating it to 50°C also changes it into different forms.¹¹ Uto et al.² were the first to make ARC in the lab (synthetic) by prenylating p-halophenols, specifically o,o’-diprenylating p-iodophenol in water and then coupling it with methyl acrylate using Mizoroki-Heck chemistry.

Figure 1: (E)-artepillin C form6Click here to View Figure

Figure 2: (Z)-artepillin C form6Click here to View Figure

The Pharmacokinetic Profile of Artipilin C

The concentration of ARC in the blood at different times has been studied, with reports indicating that it peaks approximately one hour after administration, reaching an average of 22 µg/ml. This suggests that ARC was well-absorbed and bioavailable, contributing to its significant anti-inflammatory effects.¹⁰

ARC has been shown to fight harmful free radicals and act as an effective antioxidant.^12,13 In one study, ARC at concentrations of 100 and 500 μg/ml reduced lipid peroxidation in red blood cells by 17% and 82%, respectively. Importantly, ARC exhibited low cytotoxicity, with an IC₅₀ value of 588 μM, indicating its safety at effective doses.¹³ These findings suggest that ARC has the potential to serve as a powerful dietary antioxidant and can be modified to incorporate as more potent antioxidant in dental materials.¹³

The E form of ARC was found to hinder the growth of various oral bacteria, including Streptococcus mutans, S. mitis, S. sobrinus, S. sanguinis, S. salivarius, and Lactobacillus casei, at concentrations of 200 or 400 μg/ml.¹¹ However, the breakdown products of ARC didn’t exhibit significant antibacterial effects against most strains. Interestingly, the Z form showed a relatively stronger impact on S. mitis and S. sanguinis, with MIC values of 50 and 100 μg/ml, respectively.¹¹

The anti-inflammatory effects were attributed to its inhibition of the inducible nitric oxide synthase (iNOS) gene expression by interfering with nuclear factor kappa B (NF-κB) sites in the iNOS promoter. This substance also led to a reduction in the production of prostaglandin E2 (PGE2) during instances of pain and inflammation.¹⁰ The anti-inflammatory responses of this substance were noted through modulation of the NF-κB signalling pathway.¹⁴

Potential Use in Dental Products

Artepillin C (ARC) exerts a substantial impact on dental materials (Figure 3) by targeting key mechanisms, making it a suitable candidate for incorporation into various products such as mouthwashes, toothpaste, gels, and dental restorative materials.¹⁵ The specific targets relevant to dental materials are discussed in the following sections.

Antioxidant Activity

ARC’s potent antioxidant capabilities, derived from its phenolic structure, enable it to effectively neutralize free radicals and reactive oxygen species (ROS), thereby mitigating oxidative stress and protecting cells and tissues from damage.¹⁶ These properties highlight its potential in managing oxidative stress-related oral health issues.¹⁶ Moreover, ARC has demonstrated significant DPPH radical scavenging activity, further supporting its promise as a therapeutic agent against oxidative stress-associated oral problems.¹⁶

Anti-Inflammatory Effects

ARC demonstrates anti-inflammatory properties by inhibiting pro-inflammatory molecules such as cytokines and prostaglandins.¹⁷ Its ability to suppress enzymes like cyclooxygenase (COX) suggests potential in mitigating inflammatory processes in the oral cavity, making it valuable for improving gum health and preventing periodontal diseases.¹⁷

Antimicrobial Properties

ARC, has received increased attention for its ability to inhibit the growth of microorganisms, including those that cause common oral infections^18-20. Streptococcus mutans, for example, is a primary pathogen responsible for dental caries due to its role in biofilm formation and acid production on tooth surfaces.

Porphyromonas gingivalis²⁰, a key pathogen in periodontitis, contributes to the destruction of gum tissue and bone. Candida albicans is a fungal species commonly associated with oral candidiasis, a condition prevalent among immunocompromised individuals and denture wearers^19,20.

When integrated into dental materials such as composite resins²⁰, adhesives, or cements, ARC can provide a sustained antimicrobial effect. These materials are designed to release antimicrobial agents gradually, ensuring a continuous protection against bacterial colonization and biofilm formation. This proactive inhibition of oral pathogens helps in maintaing oral hygiene, particularly in high-risk areas, such as restorations, crowns, and bridges, which are prone to secondary caries and infection^18-20.

The use of ARC in dental treatments is not just limited to reducing infections but also contributes to the longevity of dental restorations. The antibacterial properties help to preserve the underlying tooth structure, potentially reducing the need for frequent replacements of dental work, which can result from recurrent infections. As such, integrating ARC into dental materials represents a significant advancement in preventive dental care.

Immunomodulatory effects, Wound Healing, and Anti-Cancer properties

ARC has been found to influence key immune pathways, potentially enhancing the body’s ability to recognize and eliminate pathogens¹⁶. This is particularly important in the oral cavity, where the immune system constantly interacts with a complex microbiome. The modulation of immune responses can help limit chronic inflammation associated with periodontal disease and other oral infections. Through the upregulation of immune cells, such as macrophages and neutrophils, ARC can assist in quicker pathogen clearance, potentially reducing the severity and duration of infections.

In wound healing, ARC has shown promise due to its ability to stimulate the growth of fibroblasts, which are essential for tissue repair and regeneration. Fibroblasts play a crucial role in synthesizing collagen, a structural protein that forms the foundation for new tissue formation^21,22. This aspect of ARC makes it particularly valuable in oral surgery and dental procedures, such as tooth extractions, gum surgery, and implant placements, where effective wound healing is critical.

By enhancing fibroblast activity and collagen synthesis, ARC promotes faster and more efficient healing of oral tissues, reducing recovery times and the risk of complications such as infection. Furthermore, its antimicrobial properties helps in additional protection during the healing process by preventing bacterial colonization and biofilm formation on surgical wounds or dental restorations.

Tissue Regeneration

ARC, plays a significant role in tissue regeneration, particularly in periodontal therapy and guided bone regeneration (GBR). Its anti-inflammatory properties help in reducing cytokine levels, minimizing tissue damage in periodontal disease²³. ARC stimulates fibroblast activity, promoting collagen synthesis and soft tissue repair, essential for reversing gum recession. In GBR, ARC enhances osteoblast proliferation and collagen production, supporting bone regeneration. Additionally, its antimicrobial effects protect against infections, crucial during healing. ARC’s combined anti-inflammatory, regenerative, and antimicrobial properties make it highly valuable in dental procedures for rebuilding damaged tissues²³.

Biofilm Disruption

ARC effectively disrupts bacterial biofilms, including dental plaque, making it a valuable agent in enhancing oral hygiene. By inhibiting bacterial adherence, ARC prevents the formation of biofilms on teeth, particularly from Streptococcus mutans, a major contributor to dental caries²⁴. ARC also breaks down the protective matrix of established biofilms, making them easier to remove through brushing or professional cleaning. Its antimicrobial properties further reduce bacterial viability, weakening biofilms and limiting their growth. ARC’s potential in oral care products such as mouthwashes or toothpastes could help in preventing plaque build-up, reduce the risk of cavities, and support periodontal health²⁴.

Limitation

However, ARC faces challenges related to cell permeability due to its repulsion to negatively charged phospholipid-based plasma membranes²⁵. To address this, researchers have successfully esterized ARC, enhancing its cell permeability without sacrificing water solubility²⁵.Yetadditional research is needed in this direction.

Research and Development

While the potential benefits of ARC in dentistry are promising, it’s imperative to acknowledge that research in this area is ongoing. Rigorous clinical trials and further studies are essential to confirm its efficacy and safety when integrated into dental applications. Factors such as ARC concentration, compatibility with other components, and release kinetics must be meticulously examined to ensure the material’s effectiveness and safety. Regulatory approval is a critical step before these dental materials can be implemented in clinical practice.

After reviewing several articles, it is evident that there are numerous promising methods for integrating Artepillin C into diverse dental products (Figure 3)^1-20: These include,

Formulating Artepillin C-infused toothpaste.

Developing Artepillin C-based mouthwashes.

Incorporating Artepillin C into dental gels or creams.

Creating dental floss or oral strips with Artepillin C.

Designing Artepillin C-containing chewing gums for oral health benefits.

Designing dental cements with the addition of Artepillin C for enhanced properties and benefits.

Creating denture material that’s better and more beneficial by adding Artepillin C.

Figure 3: Integrating Artepillin C into diverse dental products  Click here to View Figure

After thorough research, the integration of Artepillin C into complete denture resin offers potential benefits in terms of antimicrobial, anti-inflammatory, and biocompatible properties. 

Approaches for integrating Artepillin C into dental resin encompass

Formulation Development

Research and development to create a resin formula that includes Artepillin C in optimal concentrations.

Material Compatibility

Ensuring compatibility between Artepillin C and the resin matrix to maintain the structural and mechanical integrity of the dental material.

Encapsulation Techniques

Utilizing encapsulation methods to protect the stability and release of Artepillin C within the resin matrix.

Polymerization Considerations

Adapting the curing process to accommodate the presence of Artepillin C and ensuring proper polymerization of the resin.

Testing and Validation

Conducting thorough testing to validate the antimicrobial efficacy, durability, and safety of the Artepillin C-infused resin.

Regulatory Compliance

Ensuring that the final product meets regulatory standards and guidelines for dental materials. 

Conclusion

Artepillin C from propolis emerges as a promising addition to dental materials, offering a multifaceted approach to oral health enhancement. Its antimicrobial, anti-inflammatory, and tissue-regenerative properties present exciting possibilities for innovative oral care solutions. However, thorough research and clinical validation are imperative to ascertain its safety and efficacy in dental applications, paving the way for a natural and effective approach to oral health. The ongoing exploration of ARC’s mechanisms and potential benefits underscores its significance in advancing healthcare and therapeutic interventions, emphasizing the need for continued research and exploration in this compelling field. 

Acknowledgement

The authors would like to express their gratitude to JSS Dental College and Hospital, Mysuru, Karnataka, India, and Vishnu Dental College, Bhimavaram, Andhra Pradesh, India, for providing access to their facilities, which greatly contributed to the successful completion of this study.

Funding Sources

The author(s) received no financial support for the research, authorship, and/or publication of this article.

Conflict of Interest

The author(s) do not have any conflict of interest.

Data Availability Statement

This statement does not apply to this article.

Ethics Statement

This research did not involve human participants, animal subjects, or any material that requires ethical approval.

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