Abstract
Glycyrrhiza glabra (GG) elicits protective effects against periodontal diseases. However, the sustained bioavailability of GG extract at therapeutic concentration warrants ideal delivery vehicles. Present study has focused on the design, fabrication, and evaluations of ethanolic-crude extract of GG-loaded semi-interpenetrating network (semi-IPN) hydrogel (HAAPS-GG) using alginic acid and polyvinyl alcohol (PVA) hydrogel mosaicked with HA for periodontal regeneration. The study has examined the performance of the hydrogel against the selected oral pathogens S. mutans, E. faecalis, L. acidophilus and C. albicans. HAAPS-GG was successfully fabricated and the surface functional groups were confirmed by attenuated total reflectance-infrared (ATR-IR) spectroscopy. HAAPS-GG displayed interconnecting pores, hydrophilicity and excellent water profile contributing to the biocompatibility as evident from direct contact and MTT assay in L929 fibroblasts. The hydrogel was mechanically stable and was immunocompatible owing to the relatively decreased levels of pro-inflammatory mediators COX2, 5LPO, iNOS and MPO in RAW 264.7 macrophages. In addition, the transcript analysis on RAW 264.7 revealed the down-regulation of inflammatory transcription factor NF-κβ and the pro-inflammatory cytokine TNF-α. Importantly, HAAPS-GG arrested the progression of periodontal pathogens predominantly S. mutans, and C. albicans as evident by disc diffusion assay, MTT assay and confocal microscopy. Overall, the HAAPS-GG system offers promising translational avenues in periodontal regeneration.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Availability of data and materials
Data with the raw counts matrices and annotation are available upon request from the authors through proper channels.
References
Ajagannanavar SL, Battur H, Shamarao S et al (2014) Effect of aqueous and alcoholic licorice (glycyrrhiza glabra) root extract against streptococcus mutans and lactobacillus acidophilus in comparison to chlorhexidine: an in vitro study. J Int Oral Health JIOH 4:116
Al Ahmari F, Shaikh L, Al Dhubaiban D (2020) Photodynamic therapy in the treatment of periodontal diseases: a systematic review. J Int Oral Health 12:102. https://doi.org/10.4103/jioh.jioh_204_18
Anthon GE, Barrett DM (2001) Colorimetric method for the determination of Lipoxygenase activity. J Agric Food Chem 49:32–37. https://doi.org/10.1021/jf000871s
Bahadoran M, Shamloo A, DorriNokoorani Y (2020) Development of a polyvinyl alcohol/ sodium alginate hydrogel-based scaffold incorporating bFGF-encapsulated microspheres for accelerated wound healing. Sci Rep. https://doi.org/10.1038/s41598-020-64480-9
Bayani M, Torabi S, Shahnaz A, Pourali M (2017) Main properties of nanocrystalline hydroxyapatite as a bone graft material in treatment of periodontal defects. A review of literature. Biotechnol Biotechnol Equip 31:215–220
Benov L (2019) Effect of growth media on the MTT colorimetric assay in bacteria. PLoS ONE 14:e0219713. https://doi.org/10.1371/journal.pone.0219713
Bhatia G, Khatri M, Bansal M et al (2018) A comparative evaluation of porous hydroxyapatite bone graft with and without platelet-rich plasma in the treatment of periodontal intrabony osseous defects: A clinico-radiographic study. Ind J Dent Sci 10:72. https://doi.org/10.4103/ijds.ijds_5_18
Brodbeck WG, Patel J, Voskerician G et al (2002) Biomaterial adherent macrophage apoptosis is increased by hydrophilic and anionic substrates in vivo. Proc Natl Acad Sci U S A. https://doi.org/10.1073/pnas.162124199
Chenicheri S, Usha R, Ramachandran R et al (2017) Insight into oral biofilm: Primary, secondary and residual caries and phyto-challenged solutions. Open Dent J 11:312–333. https://doi.org/10.2174/1874210601711010312
Dewi AH, Ana ID (2018) The use of hydroxyapatite bone substitute grafting for alveolar ridge preservation, sinus augmentation, and periodontal bone defect: A systematic review. Heliyon 4:e00884. https://doi.org/10.1016/j.heliyon.2018.e00884
Drury JL, Dennis RG, Mooney DJ (2004) The tensile properties of alginate hydrogels. Biomaterials. https://doi.org/10.1016/j.biomaterials.2003.10.002
Finosh GT, Jayabalan M (2015) Hybrid amphiphilic bimodal hydrogels having mechanical and biological recognition characteristics for cardiac tissue engineering. RSC Adv. https://doi.org/10.1039/c5ra04448k
Finosh GT, Jayabalan M, Vandana S, Raghu KG (2015) Hybrid alginate-polyester bimodal network hydrogel for tissue engineering—influence of structured water on long-term cellular growth. Colloids Surf B Biointerfaces 135:855–864. https://doi.org/10.1016/j.colsurfb.2015.03.020
Gasik M (2017) Understanding biomaterial-tissue interface quality: combined in vitro evaluation. Sci Technol Adv Mater 18(1):550–562. https://doi.org/10.1080/14686996.2017.1348872
GnanaprakasamThankam F, Muthu J, Sankar V, Kozhiparambil Gopal R (2013) Growth and survival of cells in biosynthetic poly vinyl alcohol-alginate IPN hydrogels for cardiac applications. Colloids Surf B Biointerfaces 107:137–145. https://doi.org/10.1016/j.colsurfb.2013.01.069
Graff G, Gamache DA, Brady MT et al (1998) Improved myeloperoxidase assay for quantitation of neutrophil influx in a rat model of endotoxin-induced uveitis. J Pharmacol Toxicol Methods 39:169–178. https://doi.org/10.1016/s1056-8719(98)00023-9
Irokawa D, Takeuchi T, Noda K et al (2017) Clinical outcome of periodontal regenerative therapy using collagen membrane and deproteinized bovine bone mineral: a 2.5-year follow-up study. BMC Res Notes 10:102. https://doi.org/10.1186/s13104-017-2426-y
Jalilzadeh-Amin G, Najarnezhad V, Anassori E et al (2015) Antiulcer properties of glycyrrhiza glabra L. Extract on experimental models of gastric ulcer in mice. Iran J Pharm Res. https://doi.org/10.22037/ijpr.2015.1752
Jasser RA, AlSubaie A, AlShehri F (2021) Effectiveness of beta-tricalcium phosphate in comparison with other materials in treating periodontal infra-bony defects around natural teeth: a systematic review and meta-analysis. BMC Oral Health 21:219. https://doi.org/10.1186/s12903-021-01570-8
Joshi D, Garg T, Goyal AK, Rath G (2016) Advanced drug delivery approaches against periodontitis. Drug Deliv 23:363–377. https://doi.org/10.3109/10717544.2014.935531
Larrañeta E, Stewart S, Ervine M et al (2018) Hydrogels for hydrophobic drug delivery. Classification, synthesis and applications. J Funct Biomater 9(1):13. https://doi.org/10.3390/jfb9010013
Li X, Shu M, Li H et al (2018) Strong, tough and mechanically self-recoverable poly(vinyl alcohol)/alginate dual-physical double-network hydrogels with large cross-link density contrast. RSC Adv 8:16674–16689. https://doi.org/10.1039/c8ra01302k
Livia Marina M, Fabricio V, Rodrigo S et al (2016) Hydroxyapatite-polycaprolactone membrane in the treatment of induced periodontal disease in dogs. Front Bioeng Biotechnol. https://doi.org/10.3389/conf.fbioe.2016.01.02373
Marnett LJ, Rowlinson SW, Goodwin DC et al (1999) Arachidonic acid oxygenation by COX-1 and COX-2: mechanism of catalysis and inhibition. J Biol Chem 274:22903–22906. https://doi.org/10.1074/jbc.274.33.22903
Matsumura K, Hyon SH, Nakajima N et al (2004) Surface modification of poly(ethylene-co-vinyl alcohol): hydroxyapatite immobilization and control of periodontal ligament. Biomaterials. https://doi.org/10.1016/j.biomaterials.2003.11.055
McMillan K, Adler M, Auld DS et al (2000) Allosteric inhibitors of inducible nitric oxide synthase dimerization discovered via combinatorial chemistry. Proc Natl Acad Sci 97:1506–1511. https://doi.org/10.1073/pnas.97.4.1506
Miranda DG, Malmonge SM, Campos DM et al (2016) A chitosan-hyaluronic acid hydrogel scaffold for periodontal tissue engineering. J Biomed Mater Res - Part B Appl Biomater. https://doi.org/10.1002/jbm.b.33516
Mohammed AA, Elsherbini AM, Ibrahim FM et al (2021) Biological effect of the nanocrystalline calcium sulfate bone graft in the periodontal regeneration. J Oral Biol Craniofacial Res 11:47–52. https://doi.org/10.1016/j.jobcr.2020.10.012
Nayak A, Laha B, Sen K (2011) Development of hydroxyapatite-ciprofloxacin bone-implants using quality by design. Acta Pharm. https://doi.org/10.2478/v10007-011-0002-x
Radhakrishnan A, Jose GM, Kurup M (2015) PEG-penetrated chitosan–alginate co-polysaccharide-based partially and fully cross-linked hydrogels as ECM mimic for tissue engineering applications. Prog Biomater. https://doi.org/10.1007/s40204-015-0041-3
Shaikh MS, Husain S, Lone MA et al (2020) Clinical effectiveness of anorganic bovine-derived hydroxyapatite matrix/cell-binding peptide grafts for regeneration of periodontal defects: a systematic review and meta-analysis. Regen Med 15:2379–2395. https://doi.org/10.2217/rme-2020-0113
Siaili M, Chatzopoulou D, Gillam DG (2018) An overview of periodontal regenerative procedures for the general dental practitioner. Saudi Dent J 30:26–37
Sidhu P, Shankargouda S, Rath A et al (2020a) Therapeutic benefits of liquorice in dentistry. J Ayurveda Integr Med 11:82–88
Szatkowski T, Kołodziejczak-Radzimska A, Zdarta J et al (2015) Synthesis and characterization of hydroxyapatite/chitosan composites. Physicochem Probl Miner Process 51:575–585. https://doi.org/10.5277/ppmp150217
Talik P, Hubicka U (2018) The DSC approach to study non-freezing water contents of hydrated hydroxypropylcellulose (HPC): a study over effects of viscosity and drug addition. J Therm Anal Calorim. https://doi.org/10.1007/s10973-017-6889-9
Teti G, Salvatore V, Focaroli S et al (2015) In vitro osteogenic and odontogenic differentiation of human dental pulp stem cells seeded on carboxymethyl cellulose-hydroxyapatite hybrid hydrogel. Front Physiol. https://doi.org/10.3389/fphys.2015.00297
Thankam FG, Muthu J (2014a) Biosynthetic hydrogels-Studies on chemical and physical characteristics on long-term cellular response for tissue engineering. J Biomed Mater Res A 102:2238–2247. https://doi.org/10.1002/jbm.a.34895
Thankam FG, Muthu J (2014b) Influence of physical and mechanical properties of amphiphilic biosynthetic hydrogels on long-term cell viability. J Mech Behav Biomed Mater 35:111–122. https://doi.org/10.1016/j.jmbbm.2014.03.010
Thankam FG, Muthu J (2015) Alginate–polyester comacromer based hydrogels as physiochemically and biologically favorable entities for cardiac tissue engineering. J Colloid Interface Sci 457:52–61. https://doi.org/10.1016/j.jcis.2015.06.034
Thankam FG, Chandra IS, Kovilam AN et al (2018) Amplification of mitochondrial activity in the healing response following rotator cuff tendon injury. Sci Rep 8:1–14. https://doi.org/10.1038/s41598-018-35391-7
Valentino A, Di Cristo F, Bosetti M et al (2021) Bioactivity and delivery strategies of phytochemical compounds in bone tissue regeneration. Appl Sci 11:5122. https://doi.org/10.3390/app11115122
Wang B, Wang J, Shao J et al (2020a) A tunable and injectable local drug delivery system for personalized periodontal application. J Cont Rel 324:134–145. https://doi.org/10.1016/j.jconrel.2020.05.004
Wang T, Zhang F, Zhao R et al (2020b) Polyvinyl alcohol/sodium alginate hydrogels incorporated with silver nanoclusters via green tea extract for antibacterial applications. Des Monomers Polym 23:118–133. https://doi.org/10.1080/15685551.2020.1804183
Yan XZ, Van Den Beucken JJJP, Cai X et al (2015) Periodontal tissue regeneration using enzymatically solidified chitosan hydrogels with or without cell loading. Tis Eng Part A. https://doi.org/10.1089/ten.tea.2014.0319
ZakeriSiavashani A, Mohammadi J, Maniura-Weber K et al (2020) Silk based scaffolds with immunomodulatory capacity: anti-inflammatory effects of nicotinic acid. Biomater Sci 8:148–162. https://doi.org/10.1039/c9bm00814d
Acknowledgements
Dr. Smitha Chenicheri is grateful to the Ms. Neenu G P and Dr. Nithin Vijayakumar and other staffs and students of CRMAS for their support to complete this study. Also, Dr. Chenicheri acknowledges Dr. P S Thaha and Dr. Sheena N of PMS College of Dental Science and Research for their encouragement for the successful completion of this study.
Funding
The authors did not receive any funding for this study.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
All the authors have read the manuscript and declare no conflict of interest. No writing assistance was utilized in the production of this manuscript.
Ethical approval
The authors have read the journal's authorship agreement and ethical policy and no animal or human research has been involved in this study.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Supplementary file 1: LSCM video showing incrreased penetration of microbes in HAAPS (AVI 7680 KB)
Supplementary file2: LSCM video showing decreased penetration of microbes in HAAPS-GG (AVI 9217 KB)
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Chenicheri, S., Ramachandran, R. & Rajamanikam, U. Antimicrobial effects of hydroxyapatite mosaicked polyvinyl alcohol-alginate semi-interpenetrating hydrogel-loaded with ethanolic extract of Glycyrrhiza glabra against oral pathogens. Prog Biomater 11, 373–383 (2022). https://doi.org/10.1007/s40204-022-00199-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s40204-022-00199-2