Biomimetic chitosan/alginate with zinc and strontium hydroxyapatite for periodontal regeneration application

Articles in Press

Document Type : Research Paper

Authors

1 Department of Chemistry, Sri Sarada Niketan College of Arts and Science for women, Salem, India

2 Department of Chemistry, Periyar University, Salem, India

3 Department of Chemistry, Government Arts and Science College Idappadi, Salem- 637102, India.

4 Vinayaka Mission's Medical College and Hospital, Vinayaka Mission's Research Foundation (Deemed to be University), Karaikal, Puducherry, India, 609609.

5 Department of Chemistry, Salem Sowdeswari College, Salem-636010.

Abstract

Objective(s): To evaluate the potential of a novel alginate/chitosan/Zinc-strontium hydroxyapatite (AG/CS/SHA) composite scaffold for periodontal tissue regeneration by assessing its biocompatibility, bioactivity, and degradation characteristics.
Materials and Methods: An AG/CS/SHA composite scaffold was fabricated using a freeze-drying technique. Characterization involved Fourier Transform Infrared Spectroscopy (FTIR) to confirm chemical composition, X-ray Diffraction (XRD) to analyze crystallinity, Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) to investigate microstructure and surface morphology. In vitro studies assessed biocompatibility using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell viability with Acridine Orange/Propidium Iodide (AO/PI) staining, alkaline phosphatase (ALP) activity, and calcium deposition to evaluate osteogenic differentiation.
Results: FTIR and XRD confirmed SHA incorporation. SEM/TEM revealed a porous structure with uniform SHA distribution. The composite exhibited controlled swelling and degradation. MTT assay and AO/PI staining demonstrated good cell viability. ALP activity (20% higher) and ARS staining (35% increase) were significantly higher in Zn and Sr in HA within AG/CS (AG/CS/SHA) scaffolds compared to AG/CS (control), indicating enhanced osteogenic differentiation and mineralization.
Conclusion: The results demonstrate that the AG/CS/SHA composite possesses favorable characteristics for bone tissue engineering applications, including excellent biocompatibility, suitable mechanical properties, and the ability to promote osteogenic differentiation. These findings suggest that the AG/CS/SHA scaffold holds significant promise as a promising biomaterial for periodontal tissue regeneration, providing a supportive environment for cell growth, differentiation, and new tissue formation.

Keywords

References

  1. Chen H, Song G, Xu T, Meng C, Zhang Y, Xin T, Yu T, Lin Y, Han B. Biomaterial Scaffolds for Periodontal Tissue Engineering. J Funct Biomater. 2024;15(8):233.
  2. Stefańska K, Volponi AA, Kulus M, Waśko J, Farzaneh M, Grzelak J, Azizidoost S, Mozdziak P, Bukowska D, Antosik P, Zabel M. Dental pulp stem cells–A basic research and future application in regenerative medicine. Biomed. Pharmacother. 2024;178:116990.
  3. Wang X, Li H, Mu M, Ye R, Zhou L, Guo G. Recent development and advances on polysaccharide composite scaffolds for dental and dentoalveolar tissue regeneration. Polym Rev. 2025;65(1):47-103.
  4. Liu JH, Bian H, Zhang Y, Long Y, Li C, Zhang R, Mao Z, Wu H, Li B, Zhi C, Lu J. Formation of Stable Zinc-Rich Amorphous Calcium Phosphate. Cryst. Growth Des. 2024 Nov 4;24(22):9492-9501.
  5. Wang CY, Chen CY, Chen KH, Lin YH, Yeh TP, Lee AK, Huang CC, Shie MY. The synergistic effects of strontium/magnesium-doped calcium silicate cement accelerates early angiogenesis and bone regeneration through double bioactive ion stimulation. Ceram Int. 2024;50(4):7121-31.
  6. Olgun O, Gül ET, Kılınç G, Gökmen F, Yıldız A, Uygur V, Sarmiento-García A. Comparative Effects of Including Inorganic, Organic, and Hydroxy Zinc Sources on Growth Development, Egg Quality, Mineral Excretion, and Bone Health of Laying Quails. Biol Trace Elem Res. 2024:1-10.
  7. Nadhiya D, Kala A, Sandhiya V, Thirunavukkarasu P, Karnan C, Prabhaharan M, Sasikumar P, Albukhaty S, Sulaiman GM. Influence of annealing temperature on structural, morphological, optical, magnetic, and antimicrobial properties of zinc ferrite nanoparticles. Plasmonics. 2024;19(4):1753-63.
  8. Lu T, Miao Y, Wu T, Ye J, Zhang Y. Effects of Zn/Sr co-substitution on the physicochemical properties and cellular responses of wollastonite. Ceram Int. 2024; 50(10):17214-17227.
  9. Robinson L, Salma-Ancane K, Stipniece L, Meenan BJ, Boyd AR. The deposition of strontium and zinc Co-substituted hydroxyapatite coatings. J Mater Sci: Mater Med. 2017;28:1-4.
  10. Hanh NT, Bich PT, Thao HT. Acute and subchronic oral toxicity assessment of calcium hydroxyapatite‐alginate in animals. Vietnam J Chem. 2019;57(1):16-20.
  11. Thang DX, Chinh NT, Ngoc DH, Tung NQ, Hoang T. Effects of processing conditions on properties and morphology of chitosan/lovastatin particles. Vietnam J Chem. 2019;57(1):85-89.
  12. Naruphontjirakul P, Li M, Boccaccini AR. Strontium and Zinc Co-Doped Mesoporous Bioactive Glass Nanoparticles for Potential Use in Bone Tissue Engineering Applications. Nanomater. 2024;14(7):575.
  13. Vijayakumar G, Sundaram GA, Mani SP, Kumar SP, Krishnan M, Lakshmanan S. Strontium and Zinc doped hydroxyapatite coating on stainless steel mini-implants used in maxillofacial surgery: An in-vitro study. Lib Pro. 2024; 44(3):1846-52.
  14. Elkhenany H, Soliman MW, Atta D, El‐Badri N. Innovative Marine‐Sourced Hydroxyapatite, Chitosan, Collagen, and Gelatin for Eco‐Friendly Bone and Cartilage Regeneration. J Biomed Mater Res Part A. 2025; 113(1):e37833.
  15. Costa WB, Félix Farias AF, Silva-Filho EC, Osajima JA, Medina-Carrasco S, Del Mar Orta M, Fonseca MG. Polysaccharide hydroxyapatite (nano) composites and their biomedical applications: An overview of recent years. ACS Omega. 2024;9(28):30035-70.
  16. Kudiyarasu S, Perumal MK, Renuka RR, Natrajan PM. Chitosan composite with mesenchymal stem cells: Properties, mechanism, and its application in bone regeneration. Int J Biol Macromol. 2024;275:133502.
  17. Ren Y, Wang Q, Xu W, Yang M, Guo W, He S, Liu W. Alginate-based hydrogels mediated biomedical applications: A review. Int J Biol Macromol. 2024; 279: 135019.
  18. Shavandi A, Bekhit AE, Ali MA, Sun Z, Gould M. Development and characterization of hydroxyapatite/β-TCP/chitosan composites for tissue engineering applications. Mater Sci Eng C. 2015; 56: 481-493.
  19. Lawrie G, Keen I, Drew B, Chandler-Temple A, Rintoul L, Fredericks P, Grøndahl L. Interactions between alginate and chitosan biopolymers characterized using FTIR and XPS. Biomacromolecules. 2007;8(8):2533-41.
  20. Haghighi M, Yazdanpanah S. Chitosan‐based coatings incorporated with cinnamon and tea extracts to extend the fish fillets shelf life: validation by FTIR spectroscopy technique. J Food Qual. 2020; 2020(1): 8865234.
  21. Li Z, Saravanakumar K, Yao L, Kim Y, Choi SY, Yoo G, Keon K, Lee CM, Youn B, Lee D, Cho N. Acer tegmentosum extract-mediated silver nanoparticles loaded chitosan/alginic acid scaffolds enhance healing of E. coli-infected wounds. International Int. J Biol Macromol. 2024;267:131389.
  22. Anaya-Sampayo LM, García-Robayo DA, Roa NS, Rodriguez-Lorenzo LM, Martínez-Cardozo C. Platelet-rich fibrin (PRF) modified nano hydroxyapatite/chitosan/gelatin/alginate scaffolds increase adhesion and viability of human dental pulp stem cells (DPSC) and osteoblasts derived from DPSC. International Int J Biol Macromol. 2024:133064.
  23. Kumar KV, Subha TJ, Ahila KG, Ravindran B, Chang SW, Mahmoud AH, Mohammed OB, Rathi MA. Spectral characterization of hydroxyapatite extracted from Black Sumatra and Fighting cock bone samples: A comparative analysis. Saudi J. Biol. Sci. 2021;28(1):840-846.
  24. Abou Taleb MF, Alkahtani A, Mohamed SK. Radiation synthesis and characterization of sodium alginate/chitosan/hydroxyapatite nanocomposite hydrogels: a drug delivery system for liver cancer. Polym Bull. 2015;72:725-42.
  25. Liu H, Cui X, Lu X, Liu X, Zhang L, Chan TS. Mechanism of Mn incorporation into hydroxyapatite: Insights from SR-XRD, Raman, XAS, and DFT calculation. Chem Geol. 2021;579:120354.
  26. Karatas E, Koc K, Yilmaz M, Aydin HM. Characterization and Comparative Investigation of Hydroxyapatite/Carboxymethyl Cellulose (CaHA/CMC) Matrix for Soft Tissue Augmentation in a Rat Model. ACS omega. 2024 Jul 12;9(29):31586-600.
  27. Abdian N, Zangbar HS, Etminanfar M, Hamishehkar H. 3D chitosan/hydroxyapatite scaffolds containing mesoporous SiO2-HA particles: A new step to healing bone defects. Int J Biol Macromol. 2024;278:135014.
  28. Perez RA, Mestres G. Role of pore size and morphology in musculo-skeletal tissue regeneration. Mater Sci Eng C. 2016;61:922-939.
  29. Monmaturapoj N, Uanlee T, Nampuksa K, Kasiwat A, Makornpan C. Preparation and properties of porous biphasic calcium phosphate/bioactive glass composite scaffolds for biomedical applications. Mater Today Commun. 2022;33:104993.
  30. Gritsch L, Perrin E, Chenal JM, Fredholm Y, Maçon AL, Chevalier J, Boccaccini AR. Combining bioresorbable polyesters and bioactive glasses: Orthopedic applications of composite implants and bone tissue engineering scaffolds. Appl Mater Today. 2021;22:100923.
  31. Abere DV, Ojo SA, Oyatogun GM, Paredes-Epinosa MB, Niluxsshun MC, Hakami A. Mechanical and morphological characterization of nano-hydroxyapatite (nHA) for bone regeneration: A mini review. Biomed Engineer Adv. 2022;4:100056.
  32. Peter M, Kumar PT, Binulal NS, Nair SV, Tamura H, Jayakumar R. Development of novel α-chitin/nanobioactive glass ceramic composite scaffolds for tissue engineering applications. Carbohydr Polym. 2009;78(4):926-31.
  33. Chen X, Wang H, Sun X, Bu Y, Yan H, Lin Q. Chemical characterization and biological properties of titania/hydroxyapatite-promoted biomimetic alginate-chitosan-gelatin composite hydrogels. Ceram Int. 2023;49(15):25744-25756.
  34. Chen X, Wu T, Bu Y, Yan H, Lin Q. Fabrication and biomedical application of alginate composite hydrogels in bone tissue engineering: A review. Int J Mol Sc. 2024; 25(14):7810.
  35. Kolanthai E, Sindu PA, Khajuria DK, Veerla SC, Kuppuswamy D, Catalani LH, Mahapatra DR. Graphene oxide—a tool for the preparation of chemically crosslinking free alginate–chitosan–collagen scaffolds for bone tissue engineering. ACS Appl Mater Interfaces. 2018;10(15):12441-12452.
  36. Shaheen TI, Montaser AS, Li S. Effect of cellulose nanocrystals on scaffolds comprising chitosan, alginate and hydroxyapatite for bone tissue engineering. Int J Biol Macromol. 2019;121:814-821.
  37. Thai NL, Beaman HT, Perlman M, Obeng EE, Du C, Monroe MB. Chitosan poly (vinyl alcohol) methacrylate hydrogels for tissue engineering scaffolds. ACS Appl Bio Mater. 2024;7(12):7818-7827.
  38. Wang F, Cai X, Shen Y, Meng L. Cell–scaffold interactions in tissue engineering for oral and craniofacial reconstruction. Bioact Mater. 2023;23:16-44.
  39. Dhivya S, Saravanan S, Sastry TP, Selvamurugan N. Nanohydroxyapatite-reinforced chitosan composite hydrogel for bone tissue repair in vitro and in vivo. J Nanobiotechnology. 2015;13:1-3.
  40. Abbass MM, El-Rashidy AA, Sadek KM, Moshy SE, Radwan IA, Rady D, Dörfer CE, Fawzy El-Sayed KM. Hydrogels and dentin–pulp complex regeneration: from the benchtop to clinical translation. Polymers. 2020;12(12):2935.
  41. Reddy MS, Ponnamma D, Choudhary R, Sadasivuni KK. A comparative review of natural and synthetic biopolymer composite scaffolds. Polymers. 2021;13(7):1105.
  42. Maadani AM, Salahinejad E. Performance comparison of PLA-and PLGA-coated porous bioceramic scaffolds: Mechanical, biodegradability, bioactivity, delivery and biocompatibility assessments. Journal of Controlled Release. 2022;351:1-7.
  43. Ghahremani-Nasab M, Akbari‑Gharalari N, Rahmani Del Bakhshayesh A, Ghotaslou A, Ebrahimi-Kalan A, Mahdipour M, Mehdipour A. Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing. Stem cell res. ther. 2023;14(1):326.
  44. Nazemoroaia M, Bagheri F, Mirahmadi-Zare SZ, Eslami-Kaliji F, Derakhshan A. Asymmetric natural wound dressing based on porous chitosan-alginate hydrogel/electrospun PCL-silk sericin loaded by 10-HDA for skin wound healing: In vitro and in vivo studies. Inter J Pharm. 2025;668:124976.
  45. He J, Hu X, Cao J, Zhang Y, Xiao J, Chen D, Xiong C, Zhang L. Chitosan-coated hydroxyapatite and drug-loaded polytrimethylene carbonate/polylactic acid scaffold for enhancing bone regeneration. Carbohydr Polym. 2021;253:117198.
  46. Owen G, Dard M, Larjava H. Hydoxyapatite/beta‐tricalcium phosphate biphasic ceramics as regenerative material for the repair of complex bone defects. J Biomed Mater Res Part B: Appl Bio Mater. 2018;106(6):2493-2512.
  47. Bose S, Tarafder S. Calcium phosphate ceramic systems in growth factor and drug delivery for bone tissue engineering: a review. Acta Biomaterialia. 2012;8(4):1401-1421.
  48. Jhundoo HD, Siefen T, Liang A, Schmidt C, Lokhnauth J, Béduneau A, Pellequer Y, Larsen CC, Lamprecht A. Anti-inflammatory activity of chitosan and 5-amino salicylic acid combinations in experimental colitis. Pharmaceutics. 2020;12(11):1038.
  49. Kim YB, Kim GH. PCL/alginate composite scaffolds for hard tissue engineering: fabrication, characterization, and cellular activities. ACS Comb Sci. 2015;17(2):87-99.
  50. Zhao DW, Liu C, Zuo KQ, Su P, Li LB, Xiao GY, Cheng L. Strontium-zinc phosphate chemical conversion coating improves the osseointegration of titanium implants by regulating macrophage polarization. Chem Eng J. 2021;408:127362.
  51. Naruphontjirakul P, Li M, Boccaccini AR. Strontium and Zinc Co-Doped Mesoporous Bioactive Glass Nanoparticles for Potential Use in Bone Tissue Engineering Applications. Nanomater. 2024;14(7):575.
Nanomedicine Journal

Articles in Press, Accepted Manuscript
Available Online from 13 May 2025

Files

Share

How to cite

Statistics