Chitosan–Aloe vera Combination Enhances STRO-1, DSPP, and Reparative Dentin Formation in a Rat Model of Reversible Pulpitis
Abstract
BACKGROUND: Direct pulp capping (DPC) aims to preserve pulp vitality but requires agents that are both bioactive and biocompatible. Calcium hydroxide (CaOH2) is widely used, however its long-term success is limited, and it may cause adverse effects. Natural biomaterials such as chitosan and Aloe vera have shown potential, but their combined regenerative effects are still not well understood. This study was conducted to evaluate the efficacy of chitosan–Aloe vera composites in promoting stem cell activation, odontoblast differentiation, and reparative dentinogenesis in a rat model of reversible pulpitis.
METHODS: Twenty-four Wistar rats with mechanically induced reversible pulpitis were divided into six groups: normal control, reversible pulpitis, CaOH2, and chitosan–Aloe vera pastes at 20%, 30%, and 40% (CA20, CA30, CA40). Pulp capping was performed following standardized pulp exposure. After 28 days, reparative dentin thickness and dentin bridge formation were assessed histologically, and STRO-1 and dentine sialophosphoprotein (DSPP) expression were analyzed immunohistochemically.
RESULTS: Dentin bridge was observed in the CA40 group, presenting the thickest dentin formation (113.5±13.5 μm). STRO-1 and DSPP were significantly higher in all chitosan–Aloe vera combination groups compared with reversible pulpitis group (p<0.01), with DSPP in CA30 and CA40 also higher than CaOH2 group. Both biomarkers demonstrated a positive correlation, and reparative dentin thickness showed a strong positive correlation with DSPP level (r=0.786, p<0.001).
CONCLUSION: Chitosan–Aloe vera combination showed encouraging biological activity in this 28-day preclinical model of reversible pulpitis. Although higher concentrations enhanced stem cell activation, odontoblast differentiation, and reparative dentin formation, these results should be interpreted cautiously due to the small sample size and study design limitations.
KEYWORDS: pulp capping, STRO-1, DSPP, dentin bridge, CaOH2, natural biomaterial, endodontics
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Sumidarti A, Moersidi SN. Reversible pulpitis accompanied with sinus tract on buccal side of the right maxillary first premolar teeth: case report. J Dentomaxillofacial Sci. 2019; 4(2): 120-3, CrossRef.
Zaen El-Din AM, Hamama HH, Abo El-Elaa MA, Grawish ME, Mahmoud SH, Neelakantan P. The effect of four materials on direct pulp capping: An animal study. Aust Endod J. 2020; 46(2): 249-56, CrossRef.
Bryniarska-Kubiak N, Basta-Kaim A, Kubiak A. Mechanobiology of dental pulp cells. Cells. 2024; 13(5): 375, CrossRef.
Xie Z, Shen Z, Zhan P, Yang J, Huang Q, Huang S, et al. Functional dental pulp regeneration: Basic research and clinical translation. Int J Mol Sci. 2021; 22(16): 8991, CrossRef.
Dieterle MP, Gross T, Steinberg T, Tomakidi P, Becker K, Vach K, et al. Characterization of a stemness-optimized purification method for human dental-pulp stem cells: An approach to standardization. Cells. 2022; 11(20): 3204, CrossRef.
Ledesma-Martínez E, Mendoza-Núñez VM, Santiago-Osorio E. Mesenchymal stem cells derived from dental pulp: A review. Stem Cells Int. 2016: 2016: 4709572, CrossRef.
Bhat A, Sirajuddin S, Prabhu SS, Chungkham S, Bilichodmath C. Iatrogenic damage to the periodontium caused by endodontic treatment procedures: An overview. Open Dent J. 2015; 9(1): 214-6, CrossRef.
Akhavan A, Arbabzadeh F, Bouzari M, Razavi SM, Davoudi A. Pulp response following direct pulp capping with dentin adhesives and mineral trioxide aggregate: An animal study. Iran Endod J. 2017; 12(2): 226-30, CrossRef.
Li Z, Cao L, Fan M, Xu Q. Direct pulp capping with calcium hydroxide or mineral trioxide aggregate: A meta-analysis. J Endod. 2015; 41(9): 1412-7, CrossRef.
Best S, Ammons CL, Karunanayake GA, Saemundsson SR, Tawil PZ. Outcome assessment of teeth with necrotic pulps and apical periodontitis treated with long-term calcium hydroxide. J Endod. 2021; 47(1): 11-8, CrossRef.
Islam R, Islam MRR, Tanaka T, Alam MK, Ahmed HMA, Sano H. Direct pulp capping procedures - Evidence and practice. Jpn Dent Sci Rev. 2023; 59: 48-61, CrossRef.
Ranjbar R, Yousefi A. Effects of aloe vera and chitosan nanoparticle thin-film membranes on wound healing in full thickness infected wounds with methicillin resistant staphylococcus aureus. Bull Emerg Trauma. 2018; 6: 8-15, CrossRef.
Erpaçal B, Adigüzel Ö, Cangül S, Acartürk M. A general overview of chitosan and its use in dentistry. Int Biol Biomed J. 2019; 5(1): 1-11, article.
Mutia MS, Sipayung YR, Elmiam D, Sihotang WY, Lina J. Granule nanoparticle citrus sinensis (L.) osbeck peel lowers blood glucose levels and HbA1C in alloxan-induced diabetes rats. Indones Biomed J. 2025; 17(1): 60-9, CrossRef.
Hamedi H, Moradi S, Hudson SM, Tonelli AE, King MW. Chitosan based bioadhesives for biomedical applications: A review. Carbohydr Polym. 2022; 282: 119100, CrossRef.
Leite YK de C, Oliveira AC de J, Quelemes PV, Neto NMA, Carvalho CES de, Soares Rodrigues HW, et al. Novel scaffold based on chitosan hydrogels/phthalated cashew gum for supporting human dental pulp stem cells. Pharmaceuticals. 2023; 16(2): 266, CrossRef.
Nugrahaningsih DAA, Purnomo E, Wasityastuti W, Martien R, Arfian N, Hartatik T. BMPR2 editing in fibroblast NIH3T3 using CRISPR/cas9 affecting BMPR2 mRNA expression and proliferation. Indones Biomed J. 2022; 14(1): 45-51, CrossRef.
Sa'adah NAM, Ardiansyah E, Fadlilah DN, Izati R, Al-Faizah BN, Kavitarna SA, et al. Green yea yogurt supplemented with L. paracasei E1 microcapsules increases erythrocyte counts and B Cell development in high-fat fructose diet mice. Mol Cell Biomed Sci. 2025; 9(1): 58-68, CrossRef.
Paul S, Modak D, Chattaraj S, Nandi D, Sarkar A, Roy J, et al. Aloe vera gel homogenate shows anti-inflammatory activity through lysosomal membrane stabilization and downregulation of TNF-α and Cox-2 gene expressions in inflammatory arthritic animals. Futur J Pharm Sci. 2021; 7(1): 1-8, CrossRef.
Sularsih. The pore size of chitosan-Aloe vera scaffold and its effect on VEGF expressions and woven alveolar bone healing of tooth extraction of Cavia cobaya. Dent J. 2020; 53(3): 115-21, CrossRef.
Sandra F. Role of herbal extract in stem cell development. Mol Cell Biomed Sci. 2018; 2(1): 19-22, CrossRef.
Widyastuti NH, Cilmiaty R, Prayitno A, Wasita B, Soetrisno S. Comparison between nanochitosane and calcium hydroxide on BMP-2 and TGF-β1 levels in pulp capping treatment (in vivo study). J Med Chem Sci. 2024; 7(5): 681-7, article.
Rashed AA, Misnan NM, Nawi MNM. Chitosan-based formulation for bone health: A review. Carbohydr Polym Technol Appl. 2023; 6(1): 100359, CrossRef.
Vu TT, Nguyen MT, Sangvanich P, Nguyen QN, Thunyakitpisal P. Acemannan used as an implantable biomaterial for vital pulp therapy of immature permanent teeth induced continued root formation. Pharmaceutics. 2020; 12(7): 644, CrossRef.
Songsiripradubboon S, Kladkaew S, Trairatvorakul C, Sangvanich P, Soontornvipart K, Banlunara W, et al. Stimulation of dentin regeneration by using acemannan in teeth with lipopolysaccharide-induced pulp inflammation. J Endod. 2017; 43(7): 1097-103, CrossRef.
Brizuela C, Ormeño A, Cabrera C, Cabezas R, Silva CI, Ramírez V, et al. Direct pulp capping with calcium hydroxide, mineral trioxide aggregate, and biodentine in permanent young teeth with caries: A randomized clinical trial. J Endod. 2017; 43(11): 1776-80, CrossRef.
Seif H, Elbanna A, Abu-Seida AM, El-Korashy DI. Regenerative potential of a novel aloe vera modified tricalcium silicate cement as a pulp capping material: An animal study. Dent Mater J. 2023; 42(6): 868-77, CrossRef.
Bedoya MA, Moreno GC, Durán C, Camacho A, Pirela AE, Rojas Lozano S, et al. Mechanically induced pulpitis: A rat model that preserves animal well-being. Biomedicines. 2025; 13(8): 1925, CrossRef.
Trongkij P, Sutimuntanakul S, Lapthanasupkul P, Chaimanakarn C, Wong R, Banomyong D. Effects of the exposure site on histological pulpal responses after direct capping with 2 calcium-silicate based cements in a rat model. Restor Dent Endod. 2018; 43(4): e36, CrossRef.
Darmadi EY, Soesilo D. Antimicrobial activity of calcium hydroxide, calcium oxide, and mineral trioxide aggregate paste against α-hemolytic streptococcus. Conserv Dent J. 2024; 14(1): 24-8, CrossRef.
Nirwana I, Munadziroh E, Yogiartono R, Thiyagu C, Ying C, DInaryanti A. Cytotoxicity and proliferation evaluation on fibroblast after combining calcium hydroxide and ellagic acid. J Adv Pharm Technol Res. 2021; 12(1): 27-31, CrossRef.
Widjiastuti I, Dewi M, Prasetyo E, Pribadi N, Moedjiono M. The cytotoxicity test of calcium hydroxide, propolis, and calcium hydroxide-propolis combination in human pulp fibroblast. J Adv Pharm Technol Res. 2020; 11(1): 20-4, CrossRef.
Alp Ş, Ulusoy N. Current approaches in pulp capping: A review. Cyprus J Med Sci. 2024; 9(3): 154-60, CrossRef.
Kaur M, Singh H, Dhillon JS, Batra M, Saini M. MTA versus biodentine: Review of literature with a comparative analysis. J Clin Diagnostic Res. 2017; 11(8): 3-7, CrossRef.
DOI: https://doi.org/10.18585/inabj.v17i6.3851
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