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17 pages, 3443 KB

Open AccessArticle

Bone Tissue Bioengineering for Craniofacial and Dental Applications: Association of Deciduous Dental Pulp Stem Cells to Carbonated Hydroxyapatite

by Nidia Silva Marinho, Carla Cristina Gomes Pinheiro, Adriana Terezinha Neves Novelino Alves, Patricia de Almeida Mattos, Jean Rodrigues Evangelista, Christian Ferreira Bernardi, José Ricardo Muniz Ferreira, Gutemberg Gomes Alves, Guilherme Frederico Bernardo Lenz e Silva, Thiago Schneider Werner Vianna, Monica Diuana Calasans-Maia, Carlos Fernando Mourão and Daniela Franco Bueno

Int. J. Mol. Sci. 2026, 27(4), 2005; https://doi.org/10.3390/ijms27042005 - 20 Feb 2026

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Abstract

Autogenous grafts remain the gold standard for repairing extensive maxillofacial bone defects, but their associated morbidity motivates the search for alternative strategies in tissue bioengineering. Deciduous dental pulp stem cells (DDPSCs) represent a promising cell source due to their accessibility, multipotency, and osteogenic [...] Read more.

Autogenous grafts remain the gold standard for repairing extensive maxillofacial bone defects, but their associated morbidity motivates the search for alternative strategies in tissue bioengineering. Deciduous dental pulp stem cells (DDPSCs) represent a promising cell source due to their accessibility, multipotency, and osteogenic potential, while nanostructured carbonated hydroxyapatite (cHA) microspheres exhibit biochemical similarity to bone mineral and favorable bioabsorption. This study investigated the osteogenic response induced by the association of DDPSCs with cHA in a rat calvaria critical-size defect model. DDPSCs were expanded, seeded onto cHA microspheres, and characterized in vitro prior to bilateral implantation in 12 Wistar rats, with each animal receiving cHA + DDPSC on the right defect and acellular cHA on the left. After 60 and 90 days, histological and histomorphometric analyses revealed new bone formation in both groups, predominantly from the defect margins toward the center. At 60 days, no significant difference in newly formed bone was observed between groups (p = 0.249). At 90 days, the DDPSC + cHA group demonstrated significantly greater bone formation compared with acellular cHA (median 40.70 vs. 11.10 histomorphometric points; p = 0.028) and significant reduction in connective tissue (p = 0.028). Complete scaffold resorption was observed in all DDPSC-treated defects at 90 days, whereas residual biomaterial persisted in the cHA group (p = 0.015), indicating progressive cHA resorption over time. These findings suggest that combining DDPSCs with cHA enhances bone regeneration and that this synthetic, bioabsorbable scaffold represents a promising strategy for future applications in bone tissue engineering. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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23 pages, 8233 KB

Open AccessArticle

Effectiveness and Safety of Mechanical Debridement for Treating Experimental Peri-Implantitis in Elderly Rats Receiving Oncological Dosages of Zoledronate

by Luan Felipe Toro, Eduardo Quintão Manhanini Souza, Vinícius Franzão Ganzaroli, Jéssica de Oliveira Alvarenga Freire, Leandro Lemes da Costa, Estevão Lopes Pereira, Beatriz Alexandrelli Machado, João Martins de Mello-Neto, Mariza Akemi Matsumoto, Cláudio Aparecido Casatti, Luciano Tavares Ângelo Cintra, Letícia Helena Theodoro, Valdir Gouveia Garcia and Edilson Ervolino

Int. J. Mol. Sci. 2026, 27(3), 1355; https://doi.org/10.3390/ijms27031355 - 29 Jan 2026

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Abstract

This study evaluated the effectiveness and safety of mechanical debridement (MD) in treating experimental peri-implantitis (EPI) in rats with osseointegrated implants, specifically those treated with high-dose zoledronate. Senescent Wistar rats underwent the extraction of their upper incisor, followed by immediate implant placement. After [...] Read more.

This study evaluated the effectiveness and safety of mechanical debridement (MD) in treating experimental peri-implantitis (EPI) in rats with osseointegrated implants, specifically those treated with high-dose zoledronate. Senescent Wistar rats underwent the extraction of their upper incisor, followed by immediate implant placement. After 8 weeks, the implants were exposed, and a transmucosal component was placed. The animals were divided into four groups: Control (C), ZOL, ZOL-EPI, and ZOL-EPI-MD. In the 9th week, drug treatment commenced, consisting of the administration of 0.45 mL of a vehicle (for group C) or zoledronate (for groups ZOL, ZOL-EPI, and ZOL-EPI-MD) every 4 days over 10 weeks. After 5 weeks of drug treatment, a cotton bandage was placed around the implants to induce EPI in the ZOL-EPI and ZOL-EPI-MD groups. In the ZOL-EPI-MD group, the ligature was removed at week 16, and local treatment was performed using MD. Euthanasia was conducted at week 19. Histological sections were obtained and stained with hematoxylin–eosin for histopathological and histometric analyses, such as the percentage of total bone tissue (B.Ar/T.Ar) and the percentage of non-vital bone tissue (NVB.Ar/B.Ar). Immunohistochemical reactions were performed to detect TNFα, IL-1β, VEGF, OCN, and TRAP. In the peri-implant connective tissue, mild, intense, and moderate inflammatory infiltrates were observed in the ZOL, ZOL-EPI, and ZOL-EPI-MD groups, respectively. Immunolabeling for TNFα and IL-1β correlated with these histopathological findings. The ZOL and ZOL-EPI-MD groups showed lower immunolabeling for VEGF compared to the control group. There was a reduction in TRAP-positive cells and lower immunolabeling for OCN in the groups treated with zoledronate, with the ZOL-EPI-MD group displaying even lower levels of OCN compared to the ZOL group. While there was no significant difference in B.Ar/T.Ar across the groups, both the ZOL, ZOL-EPI, and ZOL-EPI-MD groups exhibited higher levels of NVB.Ar/B.Ar, with the ZOL-EPI-MD group showing the highest NVB.Ar/B.Ar compared to ZOL and the other groups. In conclusion, MD, as a standalone treatment, showed neither effectiveness nor safety in the management of EPI in rats that received high doses of zoledronate. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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16 pages, 2670 KB

Open AccessArticle

The Function and Role of Intercellular Adhesion Molecule 2 in Dental Pulp Cells and Tissue

by Koudai Tashita, Daigaku Hasegawa, Yuxin Huang, He Zhao and Hidefumi Maeda

Int. J. Mol. Sci. 2025, 26(24), 12006; https://doi.org/10.3390/ijms262412006 - 13 Dec 2025

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Abstract

Direct pulp capping using mineral trioxide aggregate (MTA) is commonly used to preserve dental pulp tissue, but the molecular mechanisms underlying reparative dentin formation during this procedure and the restoration of dental pulp homeostasis remain unclear. To elucidate these mechanisms, we investigated the [...] Read more.

Direct pulp capping using mineral trioxide aggregate (MTA) is commonly used to preserve dental pulp tissue, but the molecular mechanisms underlying reparative dentin formation during this procedure and the restoration of dental pulp homeostasis remain unclear. To elucidate these mechanisms, we investigated the expression and function of intercellular adhesion molecule 2 (ICAM2) in dental pulp cells and tissue. ICAM2 expression in human dental pulp cells (HDPCs) was confirmed by the gene and protein expression analysis. ICAM2 expression during reparative dentin formation after direct pulp capping was investigated using a rat direct pulp capping model. The effect of ICAM2 on odontoblast-like differentiation of HDPCs was assessed using siRNA and magnetic cell sorting (MACS). The gene and protein expression analysis showed that ICAM2 is expressed in approximately 10% of HDPCs. Immunofluorescence staining of rat mandibular bone sections showed that ICAM2 is expressed in dental pulp tissue. During reparative dentin formation, ICAM2 expression significantly increased to nearly three-fold higher than pretreatment levels on the 3 days after direct pulp capping and then returned to normal levels. ICAM2 knockdown by siRNA enhanced odontoblast-like differentiation of HDPCs. In contrast, culture supernatant from ICAM2-positive HDPCs separated by MACS inhibited odontoblast-like differentiation of HDPCs. These findings suggest that ICAM2 may regulate reparative dentin formation in dental pulp tissue. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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20 pages, 6780 KB

Open AccessArticle

The Mitigating Effect and Mechanism of Polydeoxyribonucleotide Against Zoledronic Acid-Induced Growth Suppression of Human Gingival Fibroblasts

by Shailashree Pachhapure, Young-Min Shin, Duk Gyu Kim, Dong-Rak Choi, Jong-IL Yun, Jae-Hong Kim and Byeong-Churl Jang

Int. J. Mol. Sci. 2025, 26(23), 11367; https://doi.org/10.3390/ijms262311367 - 24 Nov 2025

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Abstract

Zoledronic acid (ZA), a nitrogen-containing bisphosphonate, is widely used to treat osteoporosis and bone metastases. However, its clinical application is limited by adverse effects, notably bisphosphonate-related osteonecrosis of the jaw (BRONJ), which is associated with cytotoxicity in oral mucosal cells. Polydeoxyribonucleotide (PDRN), a [...] Read more.

Zoledronic acid (ZA), a nitrogen-containing bisphosphonate, is widely used to treat osteoporosis and bone metastases. However, its clinical application is limited by adverse effects, notably bisphosphonate-related osteonecrosis of the jaw (BRONJ), which is associated with cytotoxicity in oral mucosal cells. Polydeoxyribonucleotide (PDRN), a salmon sperm-derived DNA polymer with regenerative and anti-inflammatory properties, has shown therapeutic potential in tissue repair; however, its ability to mitigate ZA-induced cytotoxicity remains poorly understood. Here, we investigated the molecular mechanisms of ZA-induced toxicity in HGF-1 cells, a human gingival fibroblast line, and evaluated the protective effects of PDRN. ZA treatment (50 µM, 48 h) significantly inhibited HGF-1 cell growth, accompanied by reduced phosphorylation of protein kinase B (PKB) and signal transducer and activator of transcription 3 (STAT-3), along with increased phosphorylation of TANK-binding kinase 1 (TBK1). TBK1 silencing restored cell growth under ZA exposure, whereas silencing PKB or STAT-3 further suppressed cell growth even without ZA. Co-treatment with PDRN (100 µg/mL) effectively prevented and reversed ZA-induced HGF-1 cytotoxicity. Mechanistically, PDRN inhibited ZA-induced TBK1 phosphorylation and partially restored PKB phosphorylation, though it did not reverse the reduction in p-STAT-3. Additionally, ZA significantly elevated intracellular reactive oxygen species (ROS) levels at 8 h, which were attenuated by PDRN. The antioxidant N-acetylcysteine (NAC) similarly reduced ZA-induced ROS and p-TBK1 levels and improved cell growth, although it had limited effects on p-PKB at 8 h. Importantly, delayed PDRN treatment following ZA exposure reversed ZA-induced cell growth inhibition and TBK1 activation in a dose- and time-dependent manner. In summary, these findings demonstrate that ZA suppresses HGF-1 cell growth through ROS production, TBK1 activation, and inhibition of PKB and STAT-3, whereas PDRN counteracts these effects primarily by suppressing TBK1 activation and oxidative stress. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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19 pages, 7737 KB

Open AccessArticle

Fabrication of Bioactive, 3D-Printed, Porous, Yttria-Stabilized Zirconia via Mg/Zn-Incorporated Modified Simulated Body Fluid Pretreatment

by Yuwei Wu, Shigeomi Takai and Takeshi Yabutsuka

Int. J. Mol. Sci. 2025, 26(22), 10950; https://doi.org/10.3390/ijms262210950 - 12 Nov 2025

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Abstract

Yttria-stabilized zirconia (YSZ) has attracted attention as a ceramic implant material owing to its excellent mechanical strength, biocompatibility, and aesthetic properties. However, YSZ is bioinert and lacks the ability to directly bond with bone. This study aims to enhance the bioactivity of 3D-printed [...] Read more.

Yttria-stabilized zirconia (YSZ) has attracted attention as a ceramic implant material owing to its excellent mechanical strength, biocompatibility, and aesthetic properties. However, YSZ is bioinert and lacks the ability to directly bond with bone. This study aims to enhance the bioactivity of 3D-printed porous YSZ through modified simulated body fluid (m-SBF) pretreatments. The porous YSZ substrates fabricated by fused deposition modeling were first etched with hydrofluoric acid (HF) to increase the surface roughness, followed by immersion in CO₃²⁻, Mg²⁺, and/or Zn²⁺ ion-incorporated m-SBFs. Among the tested solutions, the apatite coating formed in Mg²⁺- and Zn²⁺-containing m-SBF within one day, exhibiting uniform precipitation and a reduced tetragonal-to-monoclinic (t→m) transition. The incorporated Mg²⁺ and Zn²⁺ ions were successfully detected on the apatite coating, with Mg/Ca and Zn/Ca ratios of approximately 4.82% and 3.33%, respectively. Mg²⁺ is known to stimulate osteogenesis, while Zn²⁺ exhibits antibacterial activity. Furthermore, compared with standard SBF under high-temperature and high-pH conditions, the m-SBF induced markedly less t→m phase transition on YSZ substrates, suggesting that m-SBF, as a biomimetic medium for imparting bioactivity, provides a more suitable environment for YSZ substrates. This study demonstrates that HF surface treatment combined with Mg²⁺- and Zn²⁺-containing m-SBF pretreatment effectively imparts bioactivity to 3D-printed YSZ, offering a promising approach for next-generation osteoconductive ceramic implants. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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19 pages, 8910 KB

Open AccessArticle

Oxidized Hyaluronic Acid-Based Sponges: A Promising Biomaterial for Oral Mucosa Lesion Application

by Clara Alicia Muñoz-Trejo, Martha Gabriela Chuc-Gamboa, Juan V. Cauich-Rodríguez, Rossana Faride Vargas-Coronado, Diana María Escobar-García, Amaury Pozos-Guillen, Fernando Javier Aguilar-Pérez, Alicia Leonor Pinzón-Te and Gualberto Antonio Zumbardo-Bacelis

Int. J. Mol. Sci. 2025, 26(21), 10383; https://doi.org/10.3390/ijms262110383 - 25 Oct 2025

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Abstract

Chitosan (CHT) and hyaluronic acid (HA) are biomaterials with diverse properties. While each has been individually employed for the treatment of oral lesions, there is a need for further evidence regarding their combined properties. This study compares the effects on the properties and [...] Read more.

Chitosan (CHT) and hyaluronic acid (HA) are biomaterials with diverse properties. While each has been individually employed for the treatment of oral lesions, there is a need for further evidence regarding their combined properties. This study compares the effects on the properties and biocompatibility of chitosan sponges, CHT crosslinked with oxidized hyaluronic acid (OHA) (oxidized at 1:1 and 1:2 ratios, respectively), and CHT crosslinked with oxidized hyaluronic acid and polyethylene glycol diglycidyl ether (PEGDE). Spectroscopy revealed reduced free amino groups and the amide I/II ratio in CHT sponges crosslinked with OHA. SEM confirmed the porous network morphology with an average pore size ranging from 155 to 213 μm. TGA indicated the scaffolds’ decomposition temperature (Td) increased from 253° to 308°, with the CHT-OHA 1:2 sponge exhibiting the highest thermal stability. Compression testing highlighted that the chitosan sponges crosslinked with AHO and PEGDE at a 1:2 ratio displayed a higher elastic modulus than the other studied scaffolds. The MTS assay confirmed that the fabricated biomaterials were not cytotoxic. This study demonstrates the enhanced properties and biocompatibility of CHT-OHA and CHT-OHA-PEGDE sponges, highlighting their potential for oral lesion treatment. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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22 pages, 3187 KB

Open AccessArticle

Light-Cured Dental Fillings Containing Quinoline and Quinoxaline Derivatives: The Influence of Sorption and Solubility on Color Change—Part III

by Ilona Pyszka, Dawid Bereźnicki and Beata Jędrzejewska

Int. J. Mol. Sci. 2025, 26(19), 9537; https://doi.org/10.3390/ijms26199537 - 29 Sep 2025

Viewed by 619

Abstract

Light-cured dental fillings play a significant role in modern dentistry due to their aesthetics, durability, and ease of application. However, research is still being carried out to improve their mechanical properties, biocompatibility, and wear resistance by modifying their composition. Therefore, the aim of [...] Read more.

Light-cured dental fillings play a significant role in modern dentistry due to their aesthetics, durability, and ease of application. However, research is still being carried out to improve their mechanical properties, biocompatibility, and wear resistance by modifying their composition. Therefore, the aim of this study was to evaluate the sorption properties, solubility, and color stability of newly developed dental composites containing quinoline and quinoxaline derivatives. A total of 162 samples were prepared by mixing organic and inorganic phases. For 144 of these, color changes were assessed after conditioning in solutions simulating the oral environment, and for 18, sorption and solubility analysis were performed in distilled water according to ISO 4049. The results showed that sorption and solubility increased with increasing conditioning time. Composites characterized by higher matrix hydrophilicity showed both greater sorption and solubility as well as more pronounced color changes (ΔE > 6). In contrast, materials with lower sorption and solubility were characterized by greater color stability (ΔE < 5). The results indicate a significant impact of the hydrophilic properties of the resin matrix on the aesthetic durability of dental restorations. The data obtained can provide a basis for the appropriate design of biofunctional materials with increased resistance to degradation and discoloration. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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14 pages, 1785 KB

Open AccessArticle

Extended PRF: Impact of Heat on Gene Expression in Gingival Fibroblasts

by Xiaoyu Huang, Layla Panahipour, Dorna Rassi Faghihi, Richard J. Miron and Reinhard Gruber

Int. J. Mol. Sci. 2025, 26(18), 9120; https://doi.org/10.3390/ijms26189120 - 18 Sep 2025

Cited by 1 | Viewed by 1138

Abstract

Extended platelet-rich fibrin (e-PRF) combines the prolonged resorption properties of heat-coagulated platelet-poor plasma (PPP), becoming an albumin gel (Alb-gel) that is mixed back with the respective native cell-rich buffy coat layer (BC), i.e., concentrated PRF (C-PRF). E-PRF or Alb-PRF is utilized as a [...] Read more.

Extended platelet-rich fibrin (e-PRF) combines the prolonged resorption properties of heat-coagulated platelet-poor plasma (PPP), becoming an albumin gel (Alb-gel) that is mixed back with the respective native cell-rich buffy coat layer (BC), i.e., concentrated PRF (C-PRF). E-PRF or Alb-PRF is utilized as a barrier membrane in various clinical applications, such as guided tissue regeneration. The heating of PPP might lower its biological activity, but testing this hypothesis is necessary. To this end, we exposed gingival fibroblasts to the lysates of regular PPP, heated PPP (hPPP), and BC, followed by bulk RNA sequencing. Gingival fibroblasts responded to PPP lysates with a total of 153 up- and 71 down-regulated genes when considering a minimum 3.0-fold log2 expression change and a significance level 2.0 log-10. In sharp contrast, the response to hPPP was characterized by only five up-regulated and five down-regulated genes, clearly indicating that heating almost completely abolished the biological activity of PPP. As expected, BC was more potent than PPP and broadened the spectrum of regulated genes. RT-PCR and immunoassays confirmed the heat sensitivity of PPP as exemplified by IL11 and other genes. Moreover, PPP, but not hPPP, drives the phosphorylation of p65, representing NF-κB signaling. Taken together, these findings extend previous observations that PPP causes a robust response in gingival fibroblasts and also strengthen the hypothesis that this response is heat-sensitive. These operations support the clinical concept of e-PRF by mixing back the heated inactive PPP with the bioactive buffy coat C-PRF layer. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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17 pages, 2956 KB

Open AccessArticle

Impact of Photobiomodulation on the Pro-Osteogenic Activity of Dental Pulp Mesenchymal Stem/Stromal Cells

by Marcella Rodrigues Ueda Fernandes, Gabriella Teti, Valentina Gatta, Aurora Longhin, Ana Cecilia Corrêa Aranha and Mirella Falconi

Int. J. Mol. Sci. 2025, 26(17), 8174; https://doi.org/10.3390/ijms26178174 - 22 Aug 2025

Cited by 1 | Viewed by 1821

Abstract

Photobiomodulation (PBM) consists of applying low-level laser light to biological tissues, leading to modulation of cellular functions. PBM has recently gained much attention in the field of regenerative dentistry thanks to its powerful effect on tissue repair and regeneration. Dental pulp mesenchymal stem/stromal [...] Read more.

Photobiomodulation (PBM) consists of applying low-level laser light to biological tissues, leading to modulation of cellular functions. PBM has recently gained much attention in the field of regenerative dentistry thanks to its powerful effect on tissue repair and regeneration. Dental pulp mesenchymal stem/stromal cells (DP-MSCs) represent the ideal targets in regenerative dentistry due to their ability to stimulate the regeneration of mineralized and soft tissues and the paracrine factors that they produce. Although there have been several studies evaluating the influence of PBM on DP-MSCs’ regenerative capacity, the results are conflicting, and there are few studies on the influence of PBM on the paracrine factors released by DP-MSCs. Therefore, the aim of this study was to investigate the effect of PBM, using different energy doses of laser irradiation, on the osteogenic capacity of DP-MSCs, focusing on changes in gene expression, mineralizing ability, and release of pro-osteogenic factors. DP-MSCs were irradiated in vitro and differentiated into an osteogenic phenotype. A cell viability assay, alizarin red staining, and TEM analysis were carried out to evaluate the effect of PBM on cell activity, morphology, and mineralization ability. The expression of the main osteogenesis-related markers Runx2, Col1A1, ALP, and BMP was measured to evaluate the influence of PBM on the ability of DP-MSCs to differentiate toward an osteogenic phenotype. The release of IL-6 and IL-8, which are mainly involved in bone remodeling processes, was investigated in the cell medium following PBM irradiation. The results showed a high level of cell viability, suggesting a lack of phototoxicity under the tested conditions. Furthermore, PBM had a significant effect on mineral deposition, IL-6 and IL-8 release, and expression of osteogenic markers. TEM analysis showed intracellular modifications linked mainly to mitochondria, the endoplasmic reticulum, and autophagic vesicles after PBM treatment. These findings demonstrated that the impact of PBM on the osteogenic potential of DP-MSCs is energy dose-dependent, supporting its potential as an effective strategy in regenerative dentistry, particularly for enhancing bone remodeling. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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14 pages, 3377 KB

Open AccessArticle

Mechanical Properties of Dental Enamel in Patients with Genetic Caries Susceptibility

by Firas Haj Obeid, Karolina Jezierska, Danuta Lietz-Kijak, Piotr Skomro, Totka Bakalova, Jacek Gronwald, Piotr Baszuk, Cezary Cybulski, Wojciech Kluźniak, Barbara Gronwald, Magdalena Sroczyk-Jaszczyńska, Alicja Nowicka, Petr Louda and Helena Gronwald

Int. J. Mol. Sci. 2025, 26(16), 7749; https://doi.org/10.3390/ijms26167749 - 11 Aug 2025

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Abstract

This study evaluated the physicochemical and morphological properties of tooth enamel in patients with caries-predisposing SNPs (rs4694075 in AMBN and rs2337359 in TUFT1 genes), based on the DMFT index. We included 40 of 120 individuals (aged 19–43), collecting stimulated saliva and 58 healthy [...] Read more.

This study evaluated the physicochemical and morphological properties of tooth enamel in patients with caries-predisposing SNPs (rs4694075 in AMBN and rs2337359 in TUFT1 genes), based on the DMFT index. We included 40 of 120 individuals (aged 19–43), collecting stimulated saliva and 58 healthy teeth extracted for orthodontic/surgical reasons. Saliva DNA was genotyped. Enamel properties were assessed using Vickers microhardness, deposition thickness, and calcium content. Genotype and allele frequencies aligned with the literature. The TUFT1C/C genotype subgroup showed a significantly higher DMFT index (p = 0.03) compared to the T/T genotype, while AMBN showed no such correlation. Calcium content, microhardness, and enamel thickness were similar across all polymorphic variants of both genes. A statistically significant correlation (p = 0.003) was found between reduced enamel calcium content and a higher DMFT index. Despite existing literature on the subject, the studied SNPs did not reflect any correlation with morphological or physicochemical changes in enamel. The above results suggest that genetic variability identifies patients classified by dentists as being at higher risk of caries, even though these patients follow a non-cariogenic diet and adhere to a hygiene regime. As no structural or physicochemical changes in the enamel of this group were observed, the potential cause may be disturbances in the remineralisation mechanisms or enamel surface properties that promote biofilm adhesion in polymorphic patients. Intensive tooth calcification control algorithms using LIF and RVG, as well as remineralisation cycles to increase hydroxyapatite saturation with calcium phosphates and bioadhesive fluoride delivery systems for long-term biofilm control, are used to more effectively prevent or slow down the progression of caries. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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16 pages, 3470 KB

Open AccessArticle

Tannic Acid-Loaded Gellan Gum Hydrogels Reduce In Vitro Chemokine Expression in Oral Cells

by Natália dos Santos Sanches, Atefe Imani, Lei Wang, Otávio Augusto Pacheco Vitória, Hannah Reinert, Layla Panahipour, Francisley Ávila Souza, Idelmo Rangel Garcia Júnior and Reinhard Gruber

Int. J. Mol. Sci. 2025, 26(12), 5578; https://doi.org/10.3390/ijms26125578 - 11 Jun 2025

Cited by 2 | Viewed by 1923

Abstract

Tannic acid (TA), a natural polyphenol with antiphlogistic and crosslinking properties, is a versatile component of hydrogel that can be delivered to inflammatory sites in oral applications. However, the impact of TA dampening an inflammatory response in oral cells remains to be shown. [...] Read more.

Tannic acid (TA), a natural polyphenol with antiphlogistic and crosslinking properties, is a versatile component of hydrogel that can be delivered to inflammatory sites in oral applications. However, the impact of TA dampening an inflammatory response in oral cells remains to be shown. We, therefore, established a bioassay where chemokine expression is induced by exposing gingival fibroblasts and HSC2 oral squamous carcinoma cells to IL1β and TNFα. Additionally, gingival fibroblasts were stimulated with saliva and poly I:C HMW to trigger chemokine production. Our findings demonstrate that TA effectively reduced the expression of CXCL1, CXCL2, CXCL8, and CXCL10—in gingival fibroblasts and HSC2 cells—without affecting cell viability. This effect was further confirmed by immunoassays for CXCL8. Moreover, we observed that TA decreased ERK, JNK, and p65 phosphorylation in gingival fibroblasts and partially inhibited NF-κβ/p65 nuclear translocation. Notably, TA released from a gellan gum hydrogel retained its ability to suppress chemokine expression in gingival fibroblasts. These in vitro findings provide insights into the anti-inflammatory properties of TA in oral cells and introduce gellan gum hydrogel as a delivery vehicle paving the way for future preclinical research. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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24 pages, 3308 KB

Open AccessArticle

The Latest Achievements in the Design of Permanent Fillings for Conservative Dentistry Based on Indenoquinoxaline Derivatives as Photoinitiators of Visible-Light Polymerization: Mass and Colour Stability

by Ilona Pyszka, Oliwia Szczepańska and Beata Jędrzejewska

Int. J. Mol. Sci. 2025, 26(11), 5424; https://doi.org/10.3390/ijms26115424 - 5 Jun 2025

Viewed by 1189

Abstract

The demand for polymer composite materials in the dental market is increasing every year. This rise is due to their excellent properties and ongoing technological advancements. The goal of this study was to develop new photoinitiators included in the liquid organic matrix, which [...] Read more.

The demand for polymer composite materials in the dental market is increasing every year. This rise is due to their excellent properties and ongoing technological advancements. The goal of this study was to develop new photoinitiators included in the liquid organic matrix, which is one of the main components of dental composites. Therefore, a series of compounds based on the indenoquinoxaline skeleton was synthesized, differing in the substituent. The spectroscopic properties of these compounds allowed their use as visible-light photoinitiators of radical polymerization in combination with (phenylthio)acetic acid. In addition to the polymerization kinetics, the lifetime and quantum yield of the triplet-state formation and the rate constants of its quenching by (phenylthio)acetic acid were determined. The durability of the designed composites was also assessed. Ageing tests included hydrothermal ageing, allowing for the determination of sorption, solubility, and mass change. Solutions imitating the oral cavity environment—distilled water, artificial saliva, n-heptane, and 3% acetic acid—as well as solutions containing pigments were used for these studies. Determination of the mass change and colour stability allowed for the assessment of how these materials react to long-term exposure in the oral environment. It was found that the solution simulating the natural oral environment has a significant impact on the hydrolytic stability and colour stability of the materials. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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38 pages, 11208 KB

Open AccessArticle

Raman Spectroscopic Algorithms for Assessing Virulence in Oral Candidiasis: The Fight-or-Flight Response

by Giuseppe Pezzotti, Tetsuya Adachi, Hayata Imamura, Saki Ikegami, Ryo Kitahara, Toshiro Yamamoto, Narisato Kanamura, Wenliang Zhu, Ken-ichi Ishibashi, Kazu Okuma, Osam Mazda, Aya Komori, Hitoshi Komatsuzawa and Koichi Makimura

Int. J. Mol. Sci. 2024, 25(21), 11410; https://doi.org/10.3390/ijms252111410 - 24 Oct 2024

Cited by 3 | Viewed by 2765

Abstract

This study aimed to test the effectiveness of Raman spectroscopy in the characterization of the degrees of physiological stress and virulence in clinical swab samples collected from patients affected by oral candidiasis. Raman experiments were conducted on a series of eight isolates, both [...] Read more.

This study aimed to test the effectiveness of Raman spectroscopy in the characterization of the degrees of physiological stress and virulence in clinical swab samples collected from patients affected by oral candidiasis. Raman experiments were conducted on a series of eight isolates, both in an as-collected state and after biofilm purification followed by 3 days of culture. The outputs were matched to optical microscopy observations and the results of conventional chromogenic medium assays. A statistically significant series of ten Raman spectra were collected for each clinical sample, and their averages were examined and interpreted as multiomic snapshots for albicans and non-albicans species. Spectroscopic analyses based on selected Raman parameters previously developed for standard Candida samples revealed an extreme structural complexity for all of the clinical samples, which arose from the concurrent presence of a variety of biofilms and commensal bacteria in the samples, as well as a number of other biochemical circumstances affecting the cells in their physiological stress state. However, three Raman algorithms survived such complexity, which enabled insightful classifications of Candida cells from clinical samples, in terms of their physiological stress and morphogenic state, membrane permeability, and virulence. These three characteristics, in turn, converged into a seemingly “fight or flight” response of the Candida cells. Although yet preliminary, the present study points out criticalities and proposes solutions regarding the potential utility of Raman spectroscopy in fast bedside analyses of surveillance samples. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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19 pages, 6749 KB

Open AccessArticle

Effect of Er:YAG Pulsed Laser-Deposited Hydroxyapatite Film on Titanium Implants on M2 Macrophage Polarization In Vitro and Osteogenesis In Vivo

by Lin Ma, Min Li, Satoshi Komasa, Shigeki Hontsu, Yoshiya Hashimoto, Joji Okazaki and Kenji Maekawa

Int. J. Mol. Sci. 2024, 25(1), 349; https://doi.org/10.3390/ijms25010349 - 26 Dec 2023

Cited by 10 | Viewed by 2750

Abstract

In a previous study, we successfully coated hydroxyapatite (HAp) onto titanium (Ti) plates using the erbium-doped yttrium aluminum garnet pulsed-laser deposition (Er:YAG-PLD) method. In this study, we performed further experiments to validate the in vitro osteogenic properties, macrophage polarization, and in vivo osseointegration [...] Read more.

In a previous study, we successfully coated hydroxyapatite (HAp) onto titanium (Ti) plates using the erbium-doped yttrium aluminum garnet pulsed-laser deposition (Er:YAG-PLD) method. In this study, we performed further experiments to validate the in vitro osteogenic properties, macrophage polarization, and in vivo osseointegration activity of HAp-coated Ti (HAp-Ti) plates and screws. Briefly, we coated a HAp film onto the surfaces of Ti plates and screws via Er:YAG-PLD. The surface morphological, elemental, and crystallographic analyses confirmed the successful surface coating. The macrophage polarization and osteogenic induction were evaluated in macrophages and rat bone marrow mesenchymal stem cells, and the in vivo osteogenic properties were studied. The results showed that needle-shaped nano-HAp promoted the early expression of osteogenic and immunogenic genes in the macrophages and induced excellent M2 polarization properties. The calcium deposition and osteocalcin production were significantly higher in the HAp-Ti than in the uncoated Ti. The implantation into rat femurs revealed that the HAp-coated materials had superior osteoinductive and osseointegration activities compared with the Ti, as assessed by microcomputed tomography and histology. Thus, HAp film on sandblasted Ti plates and screws via Er:YAG-PLD enhances hard-tissue differentiation, macrophage polarization, and new bone formation in tissues surrounding implants both in vitro and in vivo. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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13 pages, 4963 KB

Open AccessArticle

Evaluation of Osteogenic Potential for Rat Adipose-Derived Stem Cells under Xeno-Free Environment

by Yuzhu Sun, Jun-Ichiro Jo and Yoshiya Hashimoto

Int. J. Mol. Sci. 2023, 24(24), 17532; https://doi.org/10.3390/ijms242417532 - 15 Dec 2023

Cited by 3 | Viewed by 1925

Abstract

This study aimed to develop a novel culture method for rat adipose-derived stem cells (rADSC) and evaluate their osteogenic potential. The rADSC cultured in xeno-free culture medium (XF-rADSCs) or conventional culture medium containing fetal bovine serum (FBS-rADSCs) were combined with micropieces of xeno-free [...] Read more.

This study aimed to develop a novel culture method for rat adipose-derived stem cells (rADSC) and evaluate their osteogenic potential. The rADSC cultured in xeno-free culture medium (XF-rADSCs) or conventional culture medium containing fetal bovine serum (FBS-rADSCs) were combined with micropieces of xeno-free recombinant collagen peptide to form 3-dimensional aggregates (XF-rADSC-CellSaic or FBS-rADSC-CellSaic). Both FBS-rADSC and XF-ADSC in CellSaic exhibited multilineage differentiation potential. Compared to FBS-rADSC-CellSaic, XF-rADSC-CellSaic accelerated and promoted osteogenic differentiation in vitro. When transplanted into rat mandibular congenital bone defects, the osteogenically differentiated XF-rADSC-CellSaic induced regeneration of bone tissue with a highly maturated structure compared to FBS-rADSC-CellSaic. In conclusion, XF-rADSC-CellSaic is a feasible 3-dimensional platform for efficient bone formation. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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11 pages, 2536 KB

Open AccessArticle

Novel Carboxylation Method for Polyetheretherketone (PEEK) Surface Modification Using Friedel–Crafts Acylation

by Xinghui Lyu, Ryuhei Kanda, Susumu Tsuda, Yoshiya Hashimoto, Takamasa Fujii and Kosuke Kashiwagi

Int. J. Mol. Sci. 2023, 24(21), 15651; https://doi.org/10.3390/ijms242115651 - 27 Oct 2023

Cited by 3 | Viewed by 2342

Abstract

Recently, polyetheretherketone (PEEK) has shown promising dental applications. Surface treatment is essential for dental applications owing to its poor surface energy and wettability; however, no consensus on an effective treatment method has been achieved. In this study, we attempted to carboxylate PEEK sample [...] Read more.

Recently, polyetheretherketone (PEEK) has shown promising dental applications. Surface treatment is essential for dental applications owing to its poor surface energy and wettability; however, no consensus on an effective treatment method has been achieved. In this study, we attempted to carboxylate PEEK sample surfaces via Friedel–Crafts acylation using succinic anhydride and AlBr₃. The possibility of further chemical modifications using carboxyl groups was examined. The samples were subjected to dehydration–condensation reactions with 1H,1H-pentadecafluorooctylamine and N,N’-dicyclohexylcarbodiimide. Furthermore, the sample’s surface properties at each reaction stage were evaluated. An absorption band in the 3300–3500 cm⁻¹ wavenumber region was observed. Additionally, peak suggestive of COOH was observed in the sample spectra. Secondary modification diminished the absorption band in 3300–3500 cm⁻¹ and a clear F1s signal was observed. Thus, Friedel–Crafts acylation with succinic anhydride produced carboxyl groups on the PEEK sample surfaces. Further chemical modification of the carboxyl groups by dehydration-condensation reactions is also possible. Thus, a series of reactions can be employed to impart desired chemical structures to PEEK surfaces. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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31 pages, 16879 KB

Open AccessArticle

In Situ Raman Study of Neurodegenerated Human Neuroblastoma Cells Exposed to Outer-Membrane Vesicles Isolated from Porphyromonas gingivalis

by Giuseppe Pezzotti, Tetsuya Adachi, Hayata Imamura, Davide Redolfi Bristol, Keiji Adachi, Toshiro Yamamoto, Narisato Kanamura, Elia Marin, Wenliang Zhu, Toshihisa Kawai, Osam Mazda, Toru Kariu, Tomonori Waku, Frank C. Nichols, Pietro Riello, Flavio Rizzolio, Tania Limongi and Kazu Okuma

Int. J. Mol. Sci. 2023, 24(17), 13351; https://doi.org/10.3390/ijms241713351 - 28 Aug 2023

Cited by 15 | Viewed by 4139

Abstract

The aim of this study was to elucidate the chemistry of cellular degeneration in human neuroblastoma cells upon exposure to outer-membrane vesicles (OMVs) produced by Porphyromonas gingivalis (Pg) oral bacteria by monitoring their metabolomic evolution using in situ Raman spectroscopy. Pg [...] Read more.

The aim of this study was to elucidate the chemistry of cellular degeneration in human neuroblastoma cells upon exposure to outer-membrane vesicles (OMVs) produced by Porphyromonas gingivalis (Pg) oral bacteria by monitoring their metabolomic evolution using in situ Raman spectroscopy. Pg-OMVs are a key factor in Alzheimer’s disease (AD) pathogenesis, as they act as efficient vectors for the delivery of toxins promoting neuronal damage. However, the chemical mechanisms underlying the direct impact of Pg-OMVs on cell metabolites at the molecular scale still remain conspicuously unclear. A widely used in vitro model employing neuroblastoma SH-SY5Y cells (a sub-line of the SK-N-SH cell line) was spectroscopically analyzed in situ before and 6 h after Pg-OMV contamination. Concurrently, Raman characterizations were also performed on isolated Pg-OMVs, which included phosphorylated dihydroceramide (PDHC) lipids and lipopolysaccharide (LPS), the latter in turn being contaminated with a highly pathogenic class of cysteine proteases, a key factor in neuronal cell degradation. Raman characterizations located lipopolysaccharide fingerprints in the vesicle structure and unveiled so far unproved aspects of the chemistry behind protein degradation induced by Pg-OMV contamination of SH-SY5Y cells. The observed alterations of cells’ Raman profiles were then discussed in view of key factors including the formation of amyloid β (Aβ) plaques and hyperphosphorylated Tau neurofibrillary tangles, and the formation of cholesterol agglomerates that exacerbate AD pathologies. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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15 pages, 26202 KB

Open AccessArticle

Bioactive Polyetheretherketone with Gelatin Hydrogel Leads to Sustained Release of Bone Morphogenetic Protein-2 and Promotes Osteogenic Differentiation

by Ruonan Zhang, Jun-Ichiro Jo, Ryuhei Kanda, Aki Nishiura, Yoshiya Hashimoto and Naoyuki Matsumoto

Int. J. Mol. Sci. 2023, 24(16), 12741; https://doi.org/10.3390/ijms241612741 - 13 Aug 2023

Cited by 14 | Viewed by 2734

Abstract

Polyetheretherketone (PEEK) is one of the most promising implant materials for hard tissues due to its similar elastic modulus; however, usage of PEEK is still limited owing to its biological inertness and low osteoconductivity. The objective of the study was to provide PEEK [...] Read more.

Polyetheretherketone (PEEK) is one of the most promising implant materials for hard tissues due to its similar elastic modulus; however, usage of PEEK is still limited owing to its biological inertness and low osteoconductivity. The objective of the study was to provide PEEK with the ability to sustain the release of growth factors and the osteogenic differentiation of stem cells. The PEEK surface was sandblasted and modified with polydopamine (PDA). Moreover, successful sandblasting and PDA modification of the PEEK surface was confirmed through physicochemical characterization. The gelatin hydrogel was then chemically bound to the PEEK by adding a solution of glutaraldehyde and gelatin to the surface of the PDA-modified PEEK. The binding and degradation of the gelatin hydrogel with PEEK (GPEEK) were confirmed, and the GPEEK mineralization was observed in simulated body fluid. Sustained release of bone morphogenetic protein (BMP)-2 was observed in GPEEK. When cultured on GPEEK with BMP-2, human mesenchymal stem cells (hMSCs) exhibited osteogenic differentiation. We conclude that PEEK with a gelatin hydrogel incorporating BMP-2 is a promising substrate for bone tissue engineering. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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Review

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16 pages, 458 KB

Open AccessReview

Leveraging Dental Biotechnology for Population Oral Health: Innovations in Prevention, Diagnosis, and Treatment

by Omer Faruk Sonmez, Thuto Serufe Makara and Raman Bedi

Int. J. Mol. Sci. 2025, 26(22), 11188; https://doi.org/10.3390/ijms262211188 - 19 Nov 2025

Cited by 2 | Viewed by 1793

Abstract

Biotechnology is reshaping dental public health by providing new tools for prevention, diagnosis, and treatment of oral diseases at scale. Salivary biomarkers enable non-invasive, early detection of caries, periodontitis, and oral cancer. Tissue engineering and regenerative approaches, driven by stem cell signaling and [...] Read more.

Biotechnology is reshaping dental public health by providing new tools for prevention, diagnosis, and treatment of oral diseases at scale. Salivary biomarkers enable non-invasive, early detection of caries, periodontitis, and oral cancer. Tissue engineering and regenerative approaches, driven by stem cell signaling and bioactive scaffolds, offer biologically integrated repair. Genomic discoveries now allow polygenic risk profiling to complement social determinants in identifying vulnerable groups, while novel biomaterials, probiotics, and vaccine research expand options for sustainable caries prevention. These innovations are underpinned by molecular mechanisms such as inflammatory signaling, stem cell differentiation pathways, and antimicrobial activity. Their translation into public health practice requires attention to affordability, regulation, equity, and workforce integration. Harnessed effectively, biotechnology can help shift oral health systems toward more preventive and equitable models of care. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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23 pages, 1957 KB

Open AccessReview

Three-Dimensional Models of the Dental Pulp: Bridging Fundamental Biology and Regenerative Therapy

by Rana Smaida, Guoqiang Hua, Nadia Benkirane-Jessel and Florence Fioretti

Int. J. Mol. Sci. 2025, 26(22), 10960; https://doi.org/10.3390/ijms262210960 - 12 Nov 2025

Cited by 1 | Viewed by 1947

Abstract

The dental pulp is a dynamic connective tissue essential for tooth vitality, sensory function, immune defense, and reparative dentinogenesis. Conventional endodontic procedures, while effective in eradicating infection, often result in a non-functional, devitalized tooth, highlighting the need for biologically based regenerative approaches. The [...] Read more.

The dental pulp is a dynamic connective tissue essential for tooth vitality, sensory function, immune defense, and reparative dentinogenesis. Conventional endodontic procedures, while effective in eradicating infection, often result in a non-functional, devitalized tooth, highlighting the need for biologically based regenerative approaches. The emergence of three-dimensional (3D) culture systems has transformed pulp biology and endodontic research by providing physiologically relevant microenvironments that better reproduce the dentino-pulp interface, vascular and neural networks, and immune interactions. This review synthesizes current advances in 3D dental pulp modeling, from scaffold-based and hydrogel systems to spheroids, organoids, bioprinted constructs, and microfluidic “tooth-on-a-chip” platforms. Each system’s composition, biological relevance, and translational potential are critically examined with respect to odontogenic differentiation, angiogenesis, neurogenesis, and inflammatory response. Applications in disease modeling, biomaterial screening, and regenerative endodontics are highlighted, showing how these models bridge fundamental biology and therapeutic innovation. Finally, we discuss key challenges including vascularization, innervation, standardization, and clinical translation, and propose integrative strategies combining bioprinting, stem-cell engineering, and organ-on-chip technologies to achieve functional pulp regeneration. Overall, 3D pulp models represent a paradigm shift from reductionist cultures to bioinstructive, patient-relevant platforms that accelerate the development of next-generation endodontic therapies. Full article

(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)

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