INTRODUCTION:

Edentulism, or edentulousness, is an irreversible condition without natural teeth that can affect the quality of life for those who have it.¹ Edentulism commonly affects elderly people and remains a major worldwide disease problem². Based on United Nations Data on World Population Aging 2019, the population of elderly with an age over 65 is estimated at 703 million people, while in Indonesia, 19% of the population suffers from edentulism, with 30% of them being elderly over 65 years old, based on the basic health research or Riset Kesehatan Dasar (Riskesdas)³. Tooth loss has an impact in various ways, including disruption of stomatognathic function, decreasing nutritional intake, and the aesthetic condition of the patient^2,4.

One of the edentulous treatments on the market is dental implant as it gives better comfort and aesthetics for the patient⁵. Besides, implants have a bone-preserving property compared to other alternative treatments⁶. The public's awareness of cosmetic dentistry has increased; hence, treatment success and comfort are better than with other alternative treatments, causing an elevation in demand for implant treatment⁷. One of the disadvantages of promoting implant treatment is the initial cost for patients, which is higher than other treatments, but compared with conventional fixed prostheses, the cumulative cost of implants is relatively lower in terms of 10 years post-treatment^7,8. Unfortunately, the high demand for medical devices does not tally with the production of local medical devices made in Indonesia.¹⁰ Generally, medical devices in Indonesia are still dominated by imported products by 97%, according to statistics released by the Indonesian Ministry of Health in 2014, including dental implants. This causes the cost of implants in Indonesia to rise with taxes, and therefore researchers are encouraged to develop implants with local materials at an affordable price¹¹.

Titanium-based implants are the most commonly used dental implants all around the world due to their biocompatibility, good mechanical properties, longevity, and high success rate^12,13. Despite the high success rate, titanium implant treatment can also cause failures¹². Allergic reactions are the most common problem with titanium implants, which happen due to the process of corrosion and wear whereby the implant release titanium ion particles in the tissue, which can trigger the inflammatory reaction and consequently disrupt the Osseointegration process^12,13. One of the alternative materials for dental implants is a composite of polymethyl methacrylate (PMMA) and hydroxyapatite (HA). PMMA is the most commonly used alloplastic biomaterial because of its good mechanical strength, relatively low price, and chemical stability¹⁴. HA is a biocompatible material that is the main ingredient in bone mineral and is easy to find. HA has osteoconductive properties that provide chemical growth of bone and tissue adhesions without causing local or systemic toxicity¹⁵. The combination of PMMA and HA increases the biocompatibility, osteoconductivity, and mechanical properties of composites¹⁶. The PMMA-HA combination provides a place for cell growth and proliferation, thus, callus formation is seen, as is the presence of osteoblasts in vertebral fractures¹⁷.

Osseointegration is used to measure the success rate of implant treatment, which is characterized by the attachment of bone to the surface of theimplant,t which eventually forms a biological seal¹⁸. The failure of the osseointegration process can occur due to poor oral hygiene, which triggers the formation of biofilms on the implant surface and results in peri implantitis¹⁹. Peri-implantitis is a pathological condition characterized by peri implant tissue inflammation and loss of supporting bone²⁰. Periodontopathogenic bacteria that often cause peri-implantitis are Gram-negative anaerobic bacteria such as Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Tannerella forsythia, Peptostreptococcus micro, Campylobacter rectus, Provotella intermedia (Pi), an Fusobacterium nucleatum (Fn).²¹ The increment of proinflammatory cytokines Interleukin 1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) are indicators of peri-implantitis where the bone matrix-forming molecules such as runt related transcription factor (RUNX2) expression are decreasing^20,21. An increase in inflammatory activity and decreased matrix-forming protein will lead to increased bone resorption activity with aincrease in these number of osteoclasts²⁰.

HA is known to have anti-biofilm and anti-bacterial activity by inhibiting the attachment of bacteria to the implant surface²². Inhibition growth and accumulation of periodontal-pathogen-caused peri-implantitis bacteria are key factors in efforts to improve dental implant success rates through the increase of the osseointegration process. Therefore, it is necessary to explore and innovate dental implant biomaterial candidates with the ability to inhibit periodontopathogenic bacteria that cause peri-implantitis at achievable prices to substitute the imported raw materials for dental implants. The purpose of this study was to analyze the periodontopathogen antibacterial properties of polymethylmethacrylate (PMMA) and hydroxyapatite (HA) composites against the growth of F. nucleatum, P. gingivalis, and A. actinomycetemcomitans in vitro.

MATERIALS AND METHODS:

Fusobacterium nucleatum culture and preparation:

Fusobacterium nucleatum (ATCC22586, UK) was cultured in TSB media and incubated for 18–24 hours at 37ᵒC under anaerobic conditions. Bacterial colonies were taken using a stick that had previously been heated with a Bunsen burner, then transferred to 3 ml of liquid Brain Heart Infusion (BHI) media and incubated at 37ᵒC for 18 hours. The bacterial suspension was equalized with the McFarland standard of 0.5 (1.5 x 10⁹ colony-forming units (CFU)/ml). The suspension is then flattened on the surface of the nutrient agar medium.

Aggregatibacter Actinomycetemcomitans Culture and Preparation: Aggregatibacter Actinomycetemcomitans (ATCC43718, UK) cultures were incubated for 24 hours at 37⁰C under anaerobic conditions in BHI media after being taken from stock using an ag sterile stick. The cultures were matched to the McFarland standard of 0.5 or the equivalent of 1.5 x 10⁸ CF/m ml. The turbidity of the bacterial suspension by double blind is equated by holding the test tubes next to each other with a white background and black stripes. The bacterial suspension is diluted once the turbidity of the bacterial suspension is not matched.

Porphyromonas gingivalis culture and preparation:

Porphyromonas gingivalis (ATCC33277, UK) was cultured in TSB media and incubated for 18–24 hours at 37ᵒC under anaerobic conditions. Bacterial colonies were taken using a stick that had previously been heated over the Bunsen burner, transferred to 3 ml of BHI liquid media, and incubated at 37ᵒC for 18 hours. The bacterial suspension was equalized with the McFarland standard of 0.5 (1.5 x 10⁹ CFU / ml). The suspension that has been equalized is taken with a micropipette and then flattened on the surface of the nutrient agar medium.

Periodontopathogen antibacterial activity of PMMA-HA Composite as Dental Implant Biomaterial Candidates: The bacterial inhibition test was carried out using the well diffusion method. Doxycycline 100 mg was used as a positive control, while there were 5 treatment groups; 1) the polymethylmethacrylate (PMMA) group (Sigma Aldrich, US), 2) the PMMA-hydroxyapatite (HA) group from the Center for Ceramics, or balai besar keramik (BBK, Indonesia), 3) the PMMA-HA group size nanoparticles (Sigma Aldrich, US); HA-BBK groups; and HA-nano groups. All samples from the treatment and positive control groups were placed on agar media that had been inoculated with bacteria and incubated for 24 hours at 37ᵒC The inhibition zone diameter was measured using a digital caliper.

Statistical Analysis: All research data were then recapitulated, analyzed descriptively, and inferentially. Data are presented as mean and standard deviation, which are presented in a bar chart. The data were analyzed using the normality and homogeneity test (p> 0.05) together with the analysis of variance (ANOVA) difference test and the post-hoc Tukey Honest Significant Difference (HSD) with a different significance value of p <0.05 using the statistical package for social science (SPSS) (IBM Corporation, Illinois, Chicago, US).

RESULT:

In this study, it was found that PMMA-HA has the ability to inhibit the growth of F. nucleatum (Figure 1). The most extensive zone of inhibition of F. nucleatum was found in doxycyline treatment, followed by HA nano, HA BBK, PMMA-HA nano, PMMA-HA BBK, and PMMA. There was a significant difference between the treatment groups in the inhibition zone of F. nucleatum. The inhibition ability of PMMA-HA nano against F. nucleatum bacteria was higher than that of PMMA-HA BBK (p = 0.0001; p <0.05).

Figure 1: Periodontopathogen antibacterial inhibition zone of PMMA-HA composites against F. nucleatum. Information: * statistically significant p-value <0.05.

The inhibition growth of A. actimomycetemcomitans shows that PMMA-HA has the highest ability to inhibit compared to others (Figure 2). The most extensive zone of inhibition of A. actimomycetemcomitans was found in doxycyline treatment, followed by HA nano, HA BBK, PMMA-HA nano, PMMA-HA BBK, and PMMA. There was a significant difference between the treatment groups in the inhibition zone of A. actimomycetemcomitans. The inhibition ability of PMMA-HA nano against A. actimomycetemcomitans was higher than that of PMMA-HA BBK (p = 0.0001; p <0.05).

Figure 2: Periodontopathogen antibacterial inhibition zone of PMMA-HA composites against A. actimomycetemcomitans. Information: * statistically significant p-value <0.05.

PMMA-HA also shows the highest ability to inhibit the growth of P. gingivalis (Figure 3). The most extensive zone of P. gingivalis inhibition was found in doxycyline treatment, followed by HA nano, PMMA-HA nano, HA BBK, PMMA-HA BBK, and PMMA. There was a significant difference between the treatment groups in the inhibition zone of P. gingivalis. The inhibition ability of PMMA-HA nano against P. gingivalis was higher than that of BBK PMMA-HA (p = 0.0001; p <0.05).

Figure 3: Periodontopathogen antibacterial inhibition zone of PMMA-HA composites against P. gingivalis. Information: * statistically significant p-value <0.05.

DISCUSSION:

Dental implants are known as a rehabilitation therapy for edentulous cases in dentistry due to their strength and stability, although their success rates have not yet met expectations²⁴. Failure in bone formation and peri-implantitis were found to be the most frequent factors playing a role in dental implant failure²⁵. This study is to develop and improve implant therapy by enhancing periodontopathogen antibacterial properties through biomaterials to corroborate the concept of osseointegration and ensure the survival and success rate of the implant. HA has been discovered and used as a promising bone tissue engineering biomaterial, including dental implants. Preceding studies discovered its capabilities as bioactive, biocompatibility, osteoinductive, and osteoconductive, which yield the osseointegration of the implant and surrounding tissues. Porous bodies, or open pore structure, and interconnected porosity of HA play a role in cell diffusion and tissue deposition²⁶. This leads to increased adhesion and adsorption of cells and plasma proteins such as fibronectin and laminin, promoting bone-to-implant contact (BIC)²⁷. Previous research using SEM analysis explicates increased and distinct filopodial attachment and adhesion of cells on the surface of HA²⁸.

HA was also found to promote the proliferation and differentiation of osteogenic mesenchymal stem cells. The preceding study conducted by real time polymerase chain reaction (RT-PCR) and immunohistochemistry analysis corroborated the osteoinductive properties of HA through the significantly increased osteogenic markers such as Runx2, ALP, osteocalcin,osteopontin,n and collagen I in peri-implant tissue up until the 3rd week compared to the control. It was also found that HA regulates downregulation of osteoclast related markers such as TNF-α, transmembrane ATPase, and TRAP, affecting bone turnover rates and osteoclastic activity^27,29,30. In addition, HA can accelerate cell mineralization through calcium ion release, which leads to the induction of the MAPK signaling pathway and promotes cell differentiation^30-32. These properties of osteoinductivity and osteoconductivity may accelerate the initiation of the osseointegration of dental implants. Albeit of its superior biological properties, HA was found to have shortcomings in clinical practice for its poor mechanical properties, as it is brittle and fragile with low mechanical strength and low fracture toughness. The development of composite technology using polymers is profoundly needed to compensate for the poor physical and mechanical capabilities of HA. Polymethyl methacrylate (PMMA) is notably one of the synthetic polymers mostly used for its bioinert and biocompatible nature. A composite of PMMA and HA was studied and proven in osseointegration for its favorable cellular adhesion, osteogenic capabilities, mechanical fixation, and strength required as a dental implant biomaterial^15,26,33.

Periimplanitis is a severe complication that can affect the implant's prognosis. Prolonged peri-implanitis can cause bone destruction, affecting the surrounding osseointegration tissue around the implant. It was used to be believed that there were similarities in the microbiota in periodontitis and peri-implantitis. However, there is a distinct microbiota differentiation between periodontitis and peri-implanitis, but there is no microbiologic marker yet for peri-implanitis. The most common bacteria in peri-implantitis were oxidized graphene nanoribbons and asaccharolytic anaerobic gram-positive rods. Some bacteria, such as A. actinomycetemcomitans, P. gingivalis, and F. nucleatum; C. albicans; Campylobacter; T. forsythia; Parvimonas micra; Staphylococcus Warneri; Pseudomonas aeruginosa; and Staphylococcus aureus, were also related to peri-implantitis. In this study, the use of A. actinomycetemcomitans, P. gingivalis, and F. nucleatum was assessed^34,35_.

The implant material is prone to becoming a place for the adhesion of bacteria and biofilm that can cause inflammatory reactions around the implant, leading to host-bacterial interaction. Bacterial virulence factors such as lipopolysaccharide (LPS) will disrupt the normal homeostasis of the body and consequently initiate inflammation, which causes the PMN to spread around the implant. Hence, the dilation of microvascular vessels and the release of pro-inflammatory cytokines such as IL-1b, IL-8a, IL-6, IL-10, IL-17, IL-21, IL-33 and TNF-α and cathepsins³⁶. This vasodilated microvascular will induce the presence of dendritic cells, macrophages, B cells, and T cells in the implant area compared with the healthy surrounding implant surface, leading to the maturation of osteoclasts and the expression of osteoblast-receptor activator nuclear kappa beta ligand (RANKL), receptor activator nuclear kappa beta (RANK), and osteoprotegrin (OPG), which results in the resorption of the bone.³⁷ Release of proinflammatory cytokine, which is induced by polymononuclear (PMN) cells, will produce reactive oxidative oxygen (ROS), enzymes, and fibroblasts that release metalloproteinase (MMP), causing degeneration of fibroblasts, which can deepen the pocket and accelerate the progression of infection. Moreover, the activity of IL-1b and IL-6 activate osteoclasts, causing tissue damage, and the activity of IL-33 can cause tissue destruction, which is correlated to periimplanitis^35,38.

Dental implant failure caused by bacterial infection could end with implant removal and a hopeless prognosis for implant re-treatment. This shows the need for antibacterial agents to preserve the dental implants. HA was found to have antibacterial properties by releasing calcium, phosphorous, and oxygen ions to the surrounding tissues, affecting cells metabolism. Recent studies have shown that the released ions would destruct the cell membrane by impairing cell permeability, penetrating the cell wall to disrupt DNA replication, and inducing ROS production, leading to cell apoptosis. These ions can also prevent the colonization of bacterial cells and the formation of biofilm by producing ROS, leading to nanotoxicity. This study has proven that the nanosized HA particle has the most prominent antibacterial activity as it releases a larger number of ions. Nanosized HA increases the surface area that is in contact with the surrounding area, thus significantly preventing antibacterial activity compared with HA-BBK. Despite the ability of the ions to produce antibacterial effects, it was found that the structure of the bacterial cell wall can also influence the degree of antibacterial activity in HA. As shown in this study, HA ions show prominent antibacterial activities as they can penetrate through the walls of A. actinomycetemcomitans, P. gingivalis, and F. nucleatum, which are gram-negative bacteria with a thin peptidoglycan wall³⁹.

HA possesses a pore structure that enables it to yield a controlled-release kinetic as one of the antibacterial activities that can prevent the growth of periodontopathogenic bacteria around the implant⁴⁰. This confirms the outstanding property of HA as an antibacterial agent, and more research is expected regarding the application of its controlled-release properties in the future. This study also found that PMMA-HA nanocomposite possesses a significant increase in bacterial inhibition compared with PMMA-HA BBK. Therefore, it can be developed as a dental implant biomaterial for clinical practices as it is proven to induce osseointegration under sufficient mechanical and antibacterial conditions.

CONCLUSION:

Based on the results of this research, it can be concluded that PMMA-HA has the ability to inhibit the growth of periodontapatogen bacteria that cause peri-implantitis, which are F. nucleatum, P. gingivalis, and A. actinomycetemcomitans. PMMA-HA nano has a better ability to inhibit the growth of F. nucleatum, P. gingivalis, andA. actinomycetemcomitans compared with PMMA-HA BBK; hence, it can be considered for dental implant biomaterials with periodontopathogenic antibacterial properties to prevent peri-implantitis. Further research is needed to study more about other periodontopathogenic bacteria causing periimplanitis and the inhibition of periodontopathogen growth by PMMA-HA in vivo for dental implants.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENTS:

The authors would like to thank the Mr. Eta Rahdianto, Research Center Airlangga University for the given facilities during the research. This study obtained Research Fund from Hibah Prioritas Riset Nasional (PRN) from Badan Riset dan Inovasi (BRIN) Republic of Indonesia in 2022 fiscal year.

REFERENCES:

1. Al-Rafee MA. The epidemiology of edentulism and the associated factors: A literature Review. J Family Med Prim Care. 2020; 9(4): 1841-1843.

2. Emami E, de Souza RF, Kabawat M, Feine JS. The impact of edentulism on oral and general health. Int J Dent. 2013; 2013: 498305.

3. Kementerian Kesehatan RI, 2019. Laporan RISKESDAS 2018. Jakarta: Lembaga Penerbit Badan Penelitian dan Pengembangan Kesehatan (LPB);197.Saskianti T, Nugraha AP, Prahasanti C, Ernawati DS, Suardita K, Riawan W. Immunohistochemical analysis of stem cells from human exfoliated deciduous teeth seeded in carbonate apatite scaffold for the alveolar bone defect in Wistar rats ( Rattus novergicus). F1000Res. 2020; 22(9): 1164

4. Sargozaie N, Moeintaghavi A, Shojaie H. Comparing the Quality of Life of Patients Requesting Dental Implants Before and After Implant. Open Dent J. 2017; Aug 31; 11: 485-491.

5. Lee DJ, Saponaro PC. Management of Edentulous Patients. Dent Clin North Am. 2019; 63(2): 249-261.

6. Atalla A. Market Analysis of Dental Care: Global Dental and Oral Care Congress. Journal of Clinical Dentistry and Oral Health.2020; 4(1): 1-2.

7. Harsono V, and Prabowo H. Dental implant as an alternative treatment for single tooth loss rehabilitation. Dentofasial. 2012; 11(3): 173.

8. Chun JS, Har A, Lim HP, Lim HJ. The analysis of cost-effectiveness of implant and conventional fixed dental prosthesis. J Adv Prosthodont. 2016; 8(1): 53-61.

9. Nazmi. Implementasi Kebijakan Pengembangan Industri Alat Kesehatan Dalam Negeri. Jurnal Kebijakan Kesehatan Indonesia. 2018; 7(1): 42-43.

10. Badan Pengkajian dan Penerapan Teknologi RI, 2020. Kurangi Dominasi Produk Impor, BPPT Luncurkan Implan Tulang Titanium Merah Putih. [online] Bppt.go.id. Available at: <https://www.bppt.go.id/teknologi-informasi-energi-dan-material/4093-kurangi-dominasi-produk-impor-bppt-luncurkan-implan-tulang-titanium-merah-putih> [Accessed 11 April 2021].

11. Kim KT, Eo MY, Nguyen TTH, Kim SM. General review of titanium toxicity. Int J Implant Dent. 2019; 5(1): 10

12. Nicholson JW. Titanium Alloys for Dental Implants: A Review. Prosthesis, 2020; 2(2): 100-116.

13. Arenas-Arrocena M, Argueta-Figueroa L, García-Contreras R, Martínez-Arenas O, Flores B, Rodriguez-Torres M, Fuente-Hernández J and Acosta-Torres L. New Trends for the Processing of Poly(Methyl Methacrylate) Biomaterial for Dental Prosthodontics. Acrylic Polymers in Healthcare. 2017. Intech Open.

14. Senra M, and Marques M. Synthetic Polymeric Materials for Bone Replacement. Journal of Composites Science. 2020; 4(4): 191.

15. Wijesinghe W, Mantilaka M, Karunarathne T, Rajapakse R. Synthesis of a hydroxyapatite/poly(methyl methacrylate) nanocomposite using dolomite. Nanoscale Advances. 2019; 1(1): 86-88.

16. Komang-Agung I, Hydravianto L, Sindrawati O, William P. Effect of Polymethylmethacrylate-Hydroxyapatite Composites on Callus Formation and Compressive Strength in Goat Vertebral Body. Malaysian Orthopaedic Journal. 2018; 12(3): 6-13.

17. Dhinakarsamy V and Jayesh R. Osseointegration. Journal of Pharmacy and Bioallied Sciences. 2015; 7(5): 228.

18. Alani A, Kelleher M, Bishop K. Peri-implantitis. Part 1: Scope of the problem. British Dental Journal. 2014; 217(6): 281-287.

19. Schminke B, vom Orde F, Gruber R, Schliephake H, Bürgers R, Miosge N. The Pathology of Bone Tissue during Peri-Implantitis. Journal of Dental Research, 2014; 94(2): 354-361.

20. Schwarz F, Derks J, Monje A, Wang H. Peri-implantitis. Journal of Periodontology. 2018; 89: S267-S290.

21. Meyer F, Enax J. Hydroxyapatite in Oral Biofilm Management. European Journal of Dentistry. 2019; 13(02): 287-290.

22. Ridwan RD Sidarningsih, Wijayanti U. The Anti-Bacterial Activity of Gingival Mucoadhesive Patch from Thymus vulgaris Essential Oil towards Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum. Research J. Pharm. and Tech. 2021; 14(2): 645-649.

23. Moraschini V, Poubel LA, Ferreira VF, Barboza Edos S. Evaluation of survival and success rates of dental implants reported in longitudinal studies with a follow-up period of at least 10 years: a systematic review. Int J Oral Maxillofac Surg. 2015; 44(3): 377-88.

24. Arcos D, Vallet-Regí M. Substituted hydroxyapatite coatings of bone implants. J Mater Chem B. 2020; 8(9): 1781-1800.

25. Gomes DS, Santos AMC, Neves GA, Menezes RR. A brief review on hydroxyapatite production and use in biomedicine. 2019; 65: 282–302.

26. Breding K, Jimbo R, Hayashi M, Xue Y, Mustafa K, Andersson M. The effect of hydroxyapatite nanocrystals on osseointegration of titanium implants: an in vivo rabbit study. Int J Dent. 2014; 2014: 171305.

27. Nayar S, Chakraverty S. A comparative study to evaluate the osteoblastic cell behavior of two nano coated titanium surfaces with NAFION stabilized the membrane. J Indian Prosthodont Soc. 2015; 15(1): 33-8.

28. Lyu LX, Zhang XF, Deegan AJ, Liang GF, Yang HN, Hu SQ, Yan XL, Huang NP, Xu T. Comparing hydroxyapatite with osteogenic medium for the osteogenic differentiation of mesenchymal stem cells on PHBV nanofibrous scaffolds. J Biomater Sci Polym Ed. 2019; 30(2): 150-161.

29. Yang X, Li Y, Liu X, Zhang R, Feng Q. In Vitro Uptake of Hydroxyapatite Nanoparticles and Their Effect on Osteogenic Differentiation of Human Mesenchymal Stem Cells. Stem Cells Int. 2018; 2018: 2036176.

30. Prahasanti C, Nugraha Ap, Saskianti T, Suardita K, Riawan W, Ernawati DS. Exfoliated Human Deciduous Tooth Stem Cells Incorporating Carbonate Apatite Scaffold Enhance BMP-2, BMP-7 and Attenuate MMP-8 Expression During Initial Alveolar Bone Remodeling in Wistar Rats (Rattus norvegicus). Clinical, Cosmetic and Investigational Dentistry 2020:12 79–85.

31. Nugraha AP, Rezkita F, Putra KG, Narmada IB, Ernawati DS, Rantam FA. Triad Tissue Engineering: Gingival Mesenchymal Stem Cells, Platelet Rich Fibrin and Hydroxyapatite Scaffold to ameliorate Relapse Post Orthodontic Treatment. Biochem. Cell. Arch. 2019; 19(2): 3689-3693.

32. Wijesinghe WPSL, Rajapaksea RMG. Synthesis of a hydroxyapatite/poly(methyl methacrylate) nanocomposite using dolomite. Nanoscale Adv.2019; 1: 86.

33. Khalil D, Hultin M. Peri-implantitis Microbiota. An Update of Dental Implantology and Biomaterial. Intech open. 2019; 84.

34. Nugraha AP, Ardani IGAW, Sitalaksmi RM, et al. Anti-Peri-implantitis Bacteria's Ability of Robusta Green Coffee Bean (Coffea Canephora) Ethanol Extract: An In Silico and In Vitro Study [published online ahead of print, 2022 Sep 8]. Eur J Dent. 2022;10.1055/s-0042-1750803. doi:10.1055/s-0042-1750803

35. Nugraha AP, Sibero MT, Nugraha AP, et al. Anti-Periodontopathogenic Ability of Mangrove Leaves (Aegiceras corniculatum) Ethanol Extract: In silico and in vitro study. Eur J Dent. 2023;17(1):46-56. doi:10.1055/s-0041-1741374

36. Nugraha AP, Ramadhani NF, Riawan W, et al. Gingival Mesenchymal Stem Cells Metabolite Decreasing TRAP, NFATc1, and Sclerostin Expression in LPS-Associated Inflammatory Osteolysis In Vivo [published online ahead of print, 2022 Jun 21]. Eur J Dent. 2022; 10.1055/s-0042-1748529. doi:10.1055/s-0042-1748529

37. Adli H. Cytokines and Peri-Implant Disease. Khazar Journal of Science and Technology. 2020; 4(1): 65-69.

38. Hajipour MJ, Fromm KM, Ashkarran AA, Jimenez de Aberasturi D, de Larramendi IR, Rojo T, Serpooshan V, Parak WJ, Mahmoudi M. Antibacterial properties of nanoparticles. Trends Biotechnol. 2012; 30(10): 499-511.

39. Ragab HS, Ibrahim FA, Abdallah F, Al-Ghamdi AA, El-Tantawy F, Radwan N, Yakuphanoglu F. Synthesis and In Vitro Antibacterial Properties of Hydroxyapatite Nanoparticles. IOSR Journal of Pharmacy and Biological Sciences. 9(1): 77–85.

40. Seyedmajidi S, Rajabnia R, Seyedmajidi M. Evaluation of antibacterial properties of hydroxyapatite/bioactive glass and fluorapatite/bioactive glass nanocomposite foams as a cellular scaffold of bone tissue. J Lab Physicians. 2018; 10(3): 265-270.

41. Parent M, Magnaudeix A, Delebassée S, Sarre E, Champion E, Viana Trecant M, Damia C. Hydroxyapatite microporous bioceramics as vancomycin reservoir: Antibacterial efficiency and biocompatibility investigation. J Biomater Appl. 2016; 31(4): 488-498.

Received on 19.04.2023 Revised on 14.01.2024

Accepted on 23.07.2024 Published on 27.03.2025

Available online from March 27, 2025

Research J. Pharmacy and Technology. 2025;18(3):1166-1171.

DOI: 10.52711/0974-360X.2025.00168

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.