Alveolar ridge preservation of two-wall bone defects using mineralized dentin matrix: An experimental pre-clinical study.

OBJECTIVES: To study bone healing of two-wall bone defects after alveolar ridge preservation using mineralized dentin matrix. MATERIALS AND METHODS: After distal roots extraction of second and fourth premolars (P2, P4) on one lateral mandible in 12 beagles, two-wall bone defects (5 × 5 × 5 mm) were surgically created distally to the remaining mesial roots of P2 and P4. A total of 24 sites were randomly allocated to three groups (implant material- time of execution): mineralized dentin matrix (MDM)-3 m (MDM + collagen membrane; 3 months), MDM-6 m (MDM particles + collagen membrane; 6 months), and C-6 m (collagen membrane only; 6 months). Clinical, radiographic, digital, and histological examinations were performed 3 and 6 months after surgery. RESULTS: The bone healing in MDM groups were better compared to Control group (volume of bone regenerated in total: 25.12 mm3 vs. 13.30 mm3, p = .046; trabecular volume/total volume: 58.84% vs. 39.18%, p = .001; new bone formation rate: 44.13% vs. 31.88%, p = .047). Vertically, the radiological bone level of bone defect in MDM-6 m group was higher than that in C-6 m group (vertical height of bone defect: 1.55 mm vs. 2.74 mm, p = .018). Horizontally, no significant differences in buccolingual bone width were found between MDM and C groups at any time or at any level below the alveolar ridge. The percentages of remaining MDM were <1% in both MDM-3 m and MDM-6 m groups. CONCLUSIONS: MDM improved bone healing of two-wall bone defects and might be considered as a socket fill material used following tooth extraction.

Biocompatibility and antimicrobial effect of demineralised dentin matrix hydrogel for dental pulp preservation.

Regeneration of dentin and preserving pulp vitality are essential targets for vital pulp therapy. Our study aimed to evaluate a novel biomimetic pulp capping agent with increased dentin regenerative activities. To produce demineralised dentin matrix (DDM) particles, human extracted teeth were ground and treated with ethylene diamine tetra-acetic acid solution. DDM particles were added to sodium alginate and this combination was dripped into a 5% calcium chloride to obtain DDM hydrogel (DDMH). The eluants of both DDMH and mineral trioxide aggregate (MTA) were tested using an MTT assay to detect their cytotoxic effect on dental pulp stem cells (DPSC). Collagen-I (COL-I) gene expression was analysed on DPSC exposed to different dilutions of pulp capping material eluants by real-time quantitative polymerase chain reaction. Acridine orange staining was used to monitor the cell growth over the tested materials. Agar diffusion assay was utilised to test the antibacterial effect of DDMH and MTA compared to controls. MTT assay revealed that neat eluates of DDMH promoted DPSC viability. However, neat eluates of MTA were cytotoxic on DPSC after 72 h of culture. Moreover, DPSC were capable of growth and attached to the surface of DDMH, while they showed a marked reduction in their number when cultured on the MTA surface for one week, as shown by the acridine orange stain. In DPSC cultured with DDMH eluates, the COL-I gene was overexpressed compared to those cultured with MTA eluants. DDMH had significant antimicrobial activity in comparison to MTA after 24 h incubation. This in vitro study showed that DDMH could be an alternative pulp capping agent for regenerative endodontics.

Revolutionizing Bone Regeneration with Grinder-Based Dentin Biomaterial: A Systematic Review.

Bone tissue regeneration is a critical aspect of dental surgery, given the common occurrence of bone resorption leading to alveolar bone defects. The aim of this paper was to conduct a systematic review to provide a comprehensive summary of the evidence regarding the regenerative properties of dentin biomaterial. This systematic review was conducted through comprehensive searches in the PubMed, Scopus, and Web of Science databases, as well as an extensive exploration of the gray literature sources, including WorldCat, The New York Academy of Medicine Library, and Trip Database, following the established PRISMA protocol. Keywords such as tooth, dentin, grinder, and autograft guided the search, with a focus on a standardized procedure involving dentin grinders within laboratory, experimental, and clinical settings. Initially, a pool of 1942 articles was identified with 452 duplicates removed. An additional 1474 articles were excluded for not aligning with the predefined topics, and three more were excluded due to the unavailability of the full text. Ultimately, 13 articles met the strict inclusion criteria and were included in the review. The chemical composition of the dentin particles was similar to natural bone in terms of oxygen, carbon, calcium, phosphorus, sodium, and magnesium content, as well as in terms of the Ca/P ratio. In addition, the dentin also contained amide I and amide II structures, as well as aliphatic and hydroxyl functional groups. The chemically treated dentin was free of microorganisms. The dentin had characteristic tubules that opened after chemical treatment. At the cellular level, dentin released bone morphogenetic protein 2, induced significant cell growth, and stimulated the reorganization of the fibroblast cytoskeleton. Most clinical studies have focused on alveolar bone regeneration. After the transplantation of demineralized dentin particles, studies have observed new bone formation, a reduction in residual bone, and an increase in connective tissue. Clinical reports consistently indicate uncomplicated healing and recovery post-transplantation. However, there is a notable gap in the evidence concerning complication rates, patient-reported outcomes, and the presence of pro-inflammatory factors. In conclusion, dentin biomaterial emerges as a versatile bone substitute, demonstrating high biocompatibility and ease of acquisition. The preservation of its internal structure containing organic matter and growth factors enhances its potential for effective bone regeneration. Particularly, in dental surgery, dentin-derived materials present a promising alternative to traditional autologous bone autografts, offering the potential to reduce patient morbidity and treatment costs.

Comparing the remineralization potential of undemineralized dentin powder versus chicken eggshell powder on artificially induced initial enamel carious lesions: an in-vitro investigation.

BACKGROUND: White spot lesions are a widespread undesirable effect, especially prevalent during fixed orthodontic treatments. The study compared the in vitro enamel remineralization potential of undemineralized dentin matrix (UDD) versus chicken eggshell powder (CESP) for artificially induced enamel lesions. METHODS: 100 caries-free and sound maxillary premolars were randomly divided into four groups each contain 25 teeth: Group I (Baseline): No treatment was done to the enamel surface. Group II (Negative control ): The enamel surface of the teeth underwent demineralization using demineralizing solution to create artificial carious lesions then kept in artificial saliva. Group III (CESP treated): After demineralizing the tooth surface, the teeth have been suspended in the CESP remineralizing solution. Group IV (UDD treated): After enamel demineralization, the teeth were suspended in UDD remineralizing solution. The remineralization potential was assessed by Vickers microhardness testing, scanning electron microscopic examination (SEM), and energy dispersive X-ray (EDX). RESULTS: The current study demonstrated an increase in the mean microhardness of CESP and UDD-treated groups; however, It was nearer to the baseline level in the UDD group. SEM imaging revealed greater enamel remineralization in the UDD group compared to the remaining groups. The UDD group disclosed complete coverage for the prismatic enamel compared to the CESP group, which revealed a partially remineralized enamel surface. Interestingly, the Ca/P ratio increased significantly in the CESP group compared to the negative control group. In contrast, a higher significant increase in the mean Ca/P ratios was recorded in the UDD group compared to the test groups. CONCLUSION: biomimetic UDD and CESP powder should be utilized to treat enamel early carious lesions. However, UDD demonstrated the most significant remineralization potential.

Histological evaluation of the regenerative potential of a novel photocrosslinkable gelatin-treated dentin matrix hydrogel in direct pulp capping: an animal study.

BACKGROUND: To assess histologically the success of the pulp capping approach performed in traumatically exposed dogs' teeth using a novel injectable gelatin-treated dentin matrix light cured hydrogel (LCG-TDM) compared with LCG, MTA and TheraCal LC. METHODS: Sixty-four dogs' teeth were divided into two groups (each including 32 teeth) based on the post-treatment evaluation period: group I: 2 weeks and group II: 8 weeks. Each group was further subdivided according to the pulp capping material into four subgroups (n = 8), with subgroup A (light-cured gelatin hydrogel) as the control subgroup, subgroup B (LCG-TDM), subgroup C (TheraCal LC), and subgroup D (MTA). Pulps were mechanically exposed in the middle of the cavity floor and capped with different materials. An assessment of periapical response was performed preoperatively and at 8 weeks. After 2 and 8-week intervals, the dogs were sacrificed, and the teeth were stained with hematoxylin-eosin and graded by using a histologic scoring system. Statistical analysis was performed using the chi-square and Kruskal-Wallis tests (p = 0.05). RESULTS: All subgroups showed mild inflammation with normal pulp tissue at 2 weeks with no significant differences between subgroups (p ≤ 0.05), except for the TheraCal LC subgroup, which exhibited moderate inflammation (62.5%). Absence of a complete calcified bridge was reported in all subgroups at 2 weeks, while at 8 weeks, the majority of samples in the LCG-TDM and MTA-Angelus subgroups showed complete dentin bridge formation and absence of inflammatory pulp response with no significant differences between them (p ≤ 0.05). However, the formed dentin in the LCG-TDM group was significantly thicker, with layers of ordered odontoblasts identified to create a homogeneous tubular structure and numerous dentinal tubule lines suggesting a favourable trend towards dentin regeneration. TheraCal LC samples revealed a reasonably thick dentin bridge with moderate inflammation (50%) and LCG showed heavily fibrous tissue infiltrates with areas of degenerated pulp with no signs of hard tissue formation. CONCLUSIONS: LCG-TDM, as an extracellular matrix-based material, has the potential to regenerate dentin and preserve pulp vitality, making it a viable natural alternative to silicate-based cements for healing in vivo dentin defects in direct pulp-capping procedures.

Comparison of clinical efficacy between autologous partially demineralized dentin matrix and deproteinized bovine bone mineral for bone augmentation in orthodontic patients with alveolar bone deficiency: a randomized controlled clinical trial.

BACKGROUND: It is common to see patients who need orthodontic treatment but with insufficient alveolar bone volume. However, safe and effective tooth movement requires sufficient alveolar bone width and height. The aim of this study is to compare the bone augmentation efficacy of Autologous Partially Demineralized Dentin Matrix (APDDM) and Deproteinized Bovine Bone Mineral (DBBM) in orthodontic patients with insufficient bone by using a randomized controlled clinical trial approach. MATERIALS AND METHODS: Twenty-seven orthodontic patients involving 40 posterior teeth alveolar sites (n = 40) with insufficient alveolar bone volume were randomly divided into a control group (n = 20) and an experimental group (n = 20). The patients in the experimental group were treated with APDDM, and those in the control group were treated with DBBM. After surgery, the adjacent teeth are moved toward the bone grafting sites according to the orthodontic treatment plan. Patients completed a postoperative response questionnaire by the Visual Analogue Scale (VAS) score to indicate pain and swelling in the bone grafted area at the time of suture removal; and CBCT scans were conducted before surgery, 6 months and 2 years after surgery to assess changes in buccal and central alveolar heights, as well as widths at the alveolar ridge apex and 3 mm, 5 mm below the apex, respectively. The CBCT image sequences were imported into Mimics 21.0 software in DICOM format. The data of the patients in both groups were collected and analyzed by SPSS 25.0. RESULTS: The VAS scores were significantly lower in the APDDM group than in the DBBM group (p < 0.05). Significant increases were observed in alveolar bone height and width at 6 months and 2 years postoperative (p < 0.05); At 2 years, the APDDM group exhibited a reduction in buccal crest height and in 3 mm, 5 mm width below alveolar ridge apex, relative to 6 months (p < 0.05), while the DBBM group showed a decrease only in the central height of the alveolar bone (p < 0.05). There was a significant bone augmentation increase found only 3 mm below the alveolar ridge apex in the APDDM group compared with the DBBM group among all 6 months group comparison (p < 0.05). At 2 years, the augmentation effects were similar across both groups (p > 0.05). CONCLUSION: Radiomics analysis indicates that APDDM serves as a viable bone augmentation material for orthodontic patients with insufficient alveolar bone volume, achieving comparable clinical efficacy to DBBM. Additionally, APDDM is associated with a milder postoperative response than DBBM. THE REGISTRATION NUMBER (TRN): ChiCTR2400084607.

Comparative Evaluation of Platelet-rich Fibrin and Treated Dentin Matrix in Regenerative Endodontic Treatment of Nonvital Immature Permanent Teeth: A Randomized Clinical Trial.

AIM: Clinical and radiographic evaluation of the efficacy of platelet-rich fibrin (PRF) and treated dentin matrix (TDM) in regenerative endodontic treatment and periapical healing of nonvital immature permanent teeth with chronic apical periodontitis. MATERIALS AND METHODS: Twenty-four children aged between 7 and 11 years, each presenting with a nonvital immature permanent upper central incisor, were selected. They were randomly allocated into two groups (n = 12), group I (PRF) and group II (TDM). Baseline clinical findings were recorded, and preoperative cone-beam computed tomography (CBCT) was taken. Follow-up was done clinically for 15 months at 3-month intervals (3, 6, 9, 12, and 15 months), and CBCT was taken at the end of the 15-month follow-up. Root length, apical diameter, radiographic root area (RRA), and size of the periapical lesion were quantitively assessed at the end of follow-up period and compared to the preoperative CBCT. RESULTS: Clinical success was 100% in both groups by the end of the follow-up period. Radiographically, after a 15-month follow-up, there was a significant increase in root length and RRA, and there was also a significant reduction in apical diameter and lesion size within each group (p < 0.05). However, there was no statistically significant difference between both groups regarding the mean percentage of increase in root length and mean percentage of reduction of apical diameter (p > 0.05). On the other hand, PRF showed more increase in RRA and more reduction in lesion size, with a statistically significant difference between both groups (p < 0.05). CONCLUSION: Both PRF and TDM were clinically successful. Platelet-rich fibrin showed better radiographic outcomes and periapical healing. CLINICAL SIGNIFICANCE: Platelet-rich fibrin is a viable scaffold to aid further root development and resolution of periapical lesions of nonvital immature permanent teeth. Further studies with different forms of TDM are needed to assess the efficacy of TDM in regenerative endodontic treatment of nonvital immature permanent teeth. How to cite this article: Asal MA, Elkalla IH, Awad SM, et al. Comparative Evaluation of Platelet-rich Fibrin and Treated Dentin Matrix in Regenerative Endodontic Treatment of Nonvital Immature Permanent Teeth: A Randomized Clinical Trial. J Contemp Dent Pract 2024;25(6):563-574.

Use of autologous dentin matrix in bone defects augmentation - a literature review.

A number of regenerative materials are currently used to regenerate or preserve the alveolar pro- cess. One of these is autogenous dentin matrix. With many valuable properties such as easy availability, simple preparation, low cost, low risk of disease transmission and no risk of triggering an immune response against the graft, autogenous dentin matrix appears to be a very good material of choice. The following article is intended to provide an overview of the use of autogenous dentin matrix.

3D-printed microgels supplemented with dentin matrix molecules as a novel biomaterial for direct pulp capping.

OBJECTIVES: To develop a 3D-printed, microparticulate hydrogel supplemented with dentin matrix molecules (DMM) as a novel regenerative strategy for dental pulp capping. MATERIALS AND METHODS: Gelatin methacryloyl microgels (7% w/v) mixed with varying concentrations of DMM were printed using a digital light projection 3D printer and lyophilized for 2 days. The release profile of the DMM-loaded microgels was measured using a bicinchoninic acid assay. Next, dental pulp exposure defects were created in maxillary first molars of Wistar rats. The exposures were randomly capped with (1) inert material - negative control, (2) microgels, (3) microgels + DMM 500 µg/ml, (4) microgels + DMM 1000 µg/ml, (5) microgels + platelet-derived growth factor (PDGF 10 ng/ml), or (6) MTA (n = 15/group). After 4 weeks, animals were euthanized, and treated molars were harvested and then processed to evaluate hard tissue deposition, pulp tissue organization, and blood vessel density. RESULTS: All the specimens from groups treated with microgel + 500 µg/ml, microgel + 1000 µg/ml, microgel + PDGF, and MTA showed the formation of organized pulp tissue, tertiary dentin, newly formed tubular and atubular dentin, and new blood vessel formation. Dentin bridge formation was greater and pulp necrosis was less in the microgel + DMM groups compared to MTA. CONCLUSIONS: The 3D-printed photocurable microgels doped with DMM exhibited favorable cellular and inflammatory pulp responses, and significantly more tertiary dentin deposition. CLINICAL RELEVANCE: 3D-printed microgel with DMM is a promising biomaterial for dentin and dental pulp regeneration in pulp capping procedures.

Photocrosslinkable gelatin-treated dentin matrix hydrogel as a novel pulp capping agent for dentin regeneration: I. synthesis, characterizations and grafting optimization.

BACKGROUND: In recent years, treated dentin matrix (TDM) has been introduced as a bioactive hydrogel for dentin regeneration in DPC. However, no study has introduced TDM as a photocrosslinkable hydrogel with a natural photoinitiating system. Therefore, the present study aimed to explore the synthesis, characterizations and grafting optimization of injectable gelatin- glycidyl methacrylate (GMA)/TDM hydrogels as a novel photocrosslinkable pulp capping agent for dentin regeneration. METHODS: G-GMA/TDM hydrogel was photocrosslinked using a new two-component photoinitiating system composed of riboflavin as a photoinitiator under visible light and glycine as a first time coinitiator with riboflavin. The grafting reaction conditions of G-GMA/TDM e.g. GMA concentration and reaction time were optimized. The kinetic parameters e.g. grafting efficiency (GE) and grafting percentage (GP%) were calculated to optimize the grafting reaction, while yield (%) was determined to monitor the formation of the hydrogel. Moreover, G-GMA/TDM hydrogels were characterized by swelling ratio, degradation degree, and cytotoxicity. The instrumental characterizations e.g. FTIR, 1H-NMR, SEM and TGA, were investigated for verifying the grafting reaction. Statistical analysis was performed using F test (ANOVA) and Post Hoc Test (P = 0.05). RESULTS: The grafting reaction dramatically increased with an increase of both GMA concentration and reaction time. It was realized that the swelling degree and degradation rate of G-GMA/TDM hydrogels were significantly reduced by increasing the GMA concentration and prolonging the reaction time. When compared to the safe low and moderate GMA content hydrogels (0.048, 0.097 M) and shorter reaction times (6, 12, 24 h), G-GMA/TDM with high GMA contents (0.195, 0.391 M) and a prolonged reaction time (48 h) demonstrated cytotoxic effects against cells using the MTT assay. Also, the morphological surface of G-GMA/TDM freeze-dried gels was found more compacted, smooth and uniform due to the grafting process. Significant thermal stability was noticed due to the grafting reaction of G-GMA/TDM throughout the TGA results. CONCLUSIONS: G-GMA/TDM composite hydrogel formed by the riboflavin/glycine photoinitiating system is a potential bioactive and biocompatible system for in-situ crosslinking the activated-light pulp capping agent for dentin regeneration.

Biodegradation of an injectable treated dentin matrix hydrogel as a novel pulp capping agent for dentin regeneration.

BACKGROUND: A novel injectable mixture termed treated dentin matrix hydrogel (TDMH) has been introduced for restoring dentin defect in DPC. However, no study evaluated its physiological biodegradation. Therefore, the present study aimed to assess scaffold homogeneity, mechanical properties and biodegradability in vitro and in vivo and the regenerated dentin induced by TDMH as a novel pulp capping agent in human permanent teeth. METHODS: Three TDMH discs were weighted, and dry/wet ratios were calculated in four slices from each disc to evaluate homogeneity. Hydrogel discs were also analyzed in triplicate to measure the compressive strength using a universal testing machine. The in vitro degradation behavior of hydrogel in PBS at 37 °C for 2 months was also investigated by monitoring the percent weight change. Moreover, 20 intact fully erupted premolars were included for assessment of TDMH in vivo biodegradation when used as a novel injectable pulp capping agent. The capped teeth were divided into four equal groups according to extraction interval after 2-, 8-, 12- and 16-weeks, stained with hematoxylin-eosin for histological and histomorphometric evaluation. Statistical analysis was performed using F test (ANOVA) and post hoc test (p = 0.05). RESULTS: No statistical differences among hydrogel slices were detected with (p = 0.192) according to homogeneity. TDMH compression modulus was (30.45 ± 1.11 kPa). Hydrogel retained its shape well up to 4 weeks and after 8 weeks completely degraded. Histological analysis after 16 weeks showed a significant reduction in TDMH area and a simultaneous significant increase in the new dentin area. The mean values of TDMH were 58.8% ± 5.9 and 9.8% ± 3.3 at 2 and 16 weeks, while the new dentin occupied 9.5% ± 2.8 at 2 weeks and 82.9% ± 3.8 at 16 weeks. CONCLUSIONS: TDMH was homogenous and exhibited significant stability and almost completely recovered after excessive compression. TDMH generally maintained their bulk geometry throughout 7 weeks. The in vivo response to TDMH was characterized by extensive degradation of the hydrogel and dentin matrix particles and abundant formation of new dentin. The degradation rate of TDMH matched the rate of new dentin formation. TRIAL REGISTRATION: PACTR201901866476410: 30/1/2019.

Marginal adaptation, physicochemical and rheological properties of treated dentin matrix hydrogel as a novel injectable pulp capping material for dentin regeneration.

BACKGROUND: Treated dentin matrix hydrogel (TDMH) has been introduced as a novel injectable direct pulp capping material. In this regard, this study aimed to evaluate its marginal adaptation, physicochemical and rheological properties for the development of clinically feasible TDMH. METHODS: TDMH was applied to the pulp floor of prepared Class I cavities (n = 5), marginal adaptation was assessed by SEM at 1000 X magnification to detect gap between dentin and filling material. Five syringes were filled with TDMH and placed between the compression plates of a universal testing machine to evaluate injectability and gelation time was also evaluated by test vial inverting method. The microstructures of lyophilized TDMH were observed by SEM. Moreover, TDMH discs (n = 5) were prepared and the water uptake (%) was determined based on the equilibrium swelling theory state of hydrogels. Its solubility was measured after one week by the ISO standard method. Rheological behaviours of TDMH (n = 5) were analysed with a rotational rheometer by computing their complex shear modulus G* and their associated storage modulus (G') and loss modulus (G''). Statistical analysis was performed using F test (ANOVA) with repeated measures and Post Hoc Test (p = 0.05). RESULTS: TDMH presented an overall 92.20 ± 2.95% of continuous margins. It exhibited gelation during the first minute, and injectability mean was 66 ± 0.36%. TDMH showed a highly porous structure, and the pores were interconnected with an average diameter about 5.09 ± 3.17 µm. Swelling equilibrium gradually reached at 6 days up to 377%. The prepared hydrogels and maintained their shape after absorbing over three times their original weight of water. TDMH fulfilled the requirements of ISO 6876, demonstrating a weight loss of 1.98 ± 0.09% and linear viscoelastic behaviour with G` 479.2 ± 12.7 and G`` 230.8 ± 13.8. CONCLUSIONS: TDMH provided good marginal adaptation, appropriate physicochemical and viscoelastic properties support its use as a novel direct pulp capping material in future clinical applications.

Evaluation of osteogenic potential of demineralized dentin matrix hydrogel for bone formation.

OBJECTIVES: Dentin, the bulk material of the tooth, resemble the bone's chemical composition and is considered a valuable bone substitute. In the current study, we assessed the cytotoxicity and osteogenic potential of demineralized dentin matrix (DDM) in comparison to HA nanoparticles (n-HA) on bone marrow mesenchymal stem cells (BMMSCs) using a hydrogel formulation. MATERIALS AND METHODS: Human extracted teeth were minced into particles and treated via chemical demineralization using ethylene diamine tetra-acetic acid solution (EDTA) to produce DDM particles. DDM and n-HA particles were added to the sodium alginate then, the combination was dripped into a 5% (w/v) calcium chloride solution to obtain DDM hydrogel (DDMH) or nano-hydroxyapatite hydrogel (NHH). The particles were evaluated by dynamic light scattering (DLS) and the hydrogels were evaluated via scanning electron microscope (SEM). BMMSCs were treated with different hydrogel concentrations (25%, 50%, 75% and neat/100%) and cell viability was evaluated using MTT assay after 72 h of culture. Collagen-I (COL-I) gene expression was studied with real-time quantitative polymerase chain reaction (RT-qPCR) after 3 weeks of culture and alkaline phosphatase (ALP) activity was assessed using enzyme-linked immune sorbent assay (ELISA) over 7th, 10th, 14th and 21st days of culture. BMMSCs seeded in a complete culture medium were used as controls. One-way ANOVA was utilized to measure the significant differences in the tested groups. RESULTS: DLS measurements revealed that DDM and n-HA particles had negative values of zeta potential. SEM micrographs showed a porous microstructure of the tested hydrogels. The viability results revealed that 100% concentrations of either DDMH or NHH were cytotoxic to BMMSCs after 72 h of culture. However, the cytotoxicity of 25% and 50% concentrations of DDMH were not statistically significant compared to the control group. RT-qPCR showed that COL-I gene expression was significantly upregulated in BMMSCs cultured with 50% DDMH compared to all other treated or control groups (P < 0.01). ELISA analysis revealed that ALP level was significantly increased in the groups treated with 50% DDMH compared to 50% NHH after 21 days in culture (P < 0.001). CONCLUSION: The injectable hydrogel containing demineralized dentin matrix was successfully formulated. DDMH has a porous structure and has been shown to provide a supporting matrix for the viability and differentiation of BMMSCs. A 50% concentration of DDMH was revealed to be not cytotoxic to BMMSCs and may have a great potential to promote bone formation ability.

Effect of locally delivered protein complex-loaded nanoparticles on bone remodelling of atrophic alveolar ridge in beagles.

OBJECTIVE: To investigate the effect of local injection of mineralized hybrid nanoparticles loading dentin matrix protein-1 (DMP-1) and matrix metalloproteinase-13 (MMP-13) complex (P-NPs) on the bone remodelling on atrophic alveolar ridges (AAR) ahead of orthodontic tooth movement (OTM). SETTINGS AND SAMPLE POPULATION: Four beagles were randomly allocated into Group C (OTM only) and Group NP (OTM with P-NPs injection). Experimental model of AAR was prepared in 8 mandibular quadrants after extraction of the third premolars (n = 4 per Group). MATERIALS AND METHODS: Reciprocal traction of the second and fourth premolars was performed towards AAR for 8 weeks. P-NPs were prepared by loading recombinant DMP-1 and MMP-13 complex into calcium carbonate (CaCO3 )-mineralized hybrid nanoparticles and injected at 0, 3 and 6 weeks. The rate of OTM and the bone remodelling characteristics were compared between Groups using fluorescent microscopic analysis and microstructural histomorphometric analysis. RESULTS: Group NP revealed higher bone volume fraction and higher trabecular ratio with lower bone mineral density than Group C on AAR area. Meanwhile, the root movement towards AAR was facilitated in Group NP representing more bodily movement than Group C. CONCLUSION: Non-invasive intervention of P-NPs injection suggested a clinical potential to facilitate translational movement into the AAR with sustaining woven bone-like microstructural environment.

Experimental study on the biocompatibility and osteogenesis induction ability of PLLA/DDM scaffolds.

To investigate the characterization and function of a novel porous osteogenic material (PLLA / DDM) containing polylactic acid and demineralized dentin matrix. The surface morphology and composition of the material were observed by SEM and EDS. The physical characteristics of the material were detected by roughness and water contact angle analyses. The rate of weight loss and change in the pH value of the material were observed by scaffold degradation experiments. Four types of material were investigated: polylactic acid (PLLA) scaffold, demineralized dentin matrix (DDM) particles, PLLA/DDM scaffold and a blank control. The osteogenic effect and osteogenic characteristics of the new materials were explored through in vivo and in vitro osteogenic experiments. SEM analysis showed that DDM powder was uniformly distributed in the polylactic acid scaffold, and the water contact angle revealed that the water absorption of the porous scaffold was improved after the addition of DDM powder. The EDS results showed that the peak values of calcium and phosphorus were obviously increased after the addition of DDM powder, and the porosity test showed that the scaffold had higher porosity after the addition of DDM powder. Scaffold degradation experiments revealed that the scaffold gradually degraded with increasing time, and its pH value slightly increased. The results of cell culture and animal model experiments showed that the porous PLLA/DDM scaffold had good bio-compatibility and promoted cell proliferation and differentiation. In histological and micro-CT evaluations, the material showed good bio-compatibility, biodegradability and bone conductivity with host bone tissue in vivo. PLLA / DDM hybrid showed better performance than PLLA or DDM. The biocompatibility and cell growth promoting properties were stronger than those of single material.

Human Amnion Epithelial Cells: A Potential Cell Source for Pulp Regeneration?

The aim of this study was to analyze the suitability of pluripotent stem cells derived from the amnion (hAECs) as a potential cell source for revitalization in vitro. hAECs were isolated from human placentas, and dental pulp stem cells (hDPSCs) and dentin matrix proteins (eDMPs) were obtained from human teeth. Both hAECs and hDPSCs were cultured with 10% FBS, eDMPs and an osteogenic differentiation medium (StemPro). Viability was assessed by MTT and cell adherence to dentin was evaluated by scanning electron microscopy. Furthermore, the expression of mineralization-, odontogenic differentiation- and epithelial-mesenchymal transition-associated genes was analyzed by quantitative real-time PCR, and mineralization was evaluated through Alizarin Red staining. The viability of hAECs was significantly lower compared with hDPSCs in all groups and at all time points. Both hAECs and hDPSCs adhered to dentin and were homogeneously distributed. The regulation of odontoblast differentiation- and mineralization-associated genes showed the lack of transition of hAECs into an odontoblastic phenotype; however, genes associated with epithelial-mesenchymal transition were significantly upregulated in hAECs. hAECs showed small amounts of calcium deposition after osteogenic differentiation with StemPro. Pluripotent hAECs adhere on dentin and possess the capacity to mineralize. However, they presented an unfavorable proliferation behavior and failed to undergo odontoblastic transition.

Perivascular osteoprogenitors are associated with transcortical channels of long bones.

Bone remodeling and regeneration are dependent on resident stem/progenitor cells with the ability to replenish mature osteoblasts and repair the skeleton. Using lineage tracing approaches, we identified a population of Dmp1+ cells that reside within cortical bone and are distinct from osteocytes. Our aims were to characterize this stromal population of transcortical perivascular cells (TPCs) in their resident niche and evaluate their osteogenic potential. To distinguish this population from osteoblasts/osteocytes, we crossed mice containing inducible DMP1CreERT2/Ai9 Tomato reporter (iDMP/T) with Col2.3GFP reporter (ColGFP), a marker of osteoblasts and osteocytes. We observed iDMP/T+;ColGFP- TPCs within cortical bone following tamoxifen injection. These cells were perivascular and located within transcortical channels. Ex vivo bone outgrowth cultures showed TPCs migrated out of the channels onto the plate and expressed stem cell markers such as Sca1, platelet derived growth factor receptor beta (PDGFRß), and leptin receptor. In a cortical bone transplantation model, TPCs migrate from their vascular niche within cortical bone and contribute to new osteoblast formation and bone tube closure. Treatment with intermittent parathyroid hormone increased TPC number and differentiation. TPCs were unable to differentiate into adipocytes in the presence of rosiglitazone in vitro or in vivo. Altogether, we have identified and characterized a novel stromal lineage-restricted osteoprogenitor that is associated with transcortical vessels of long bones. Functionally, we have demonstrated that this population can migrate out of cortical bone channels, expand, and differentiate into osteoblasts, therefore serving as a source of progenitors contributing to new bone formation.

DMP-1 promoter-associated antisense strand non-coding RNA, panRNA-DMP-1, physically associates with EGFR to repress EGF-induced squamous cell carcinoma migration.

Accumulating evidence suggests that specific non-coding RNAs exist in many types of malignant tissues, and are involved in cancer invasion and metastasis. However, little is known about the precise roles of non-coding RNAs in squamous cell carcinoma (SQCC) invasion and migration. Recently, the dentin matrix protein-1 (DMP-1) gene locus was identified as a transcriptionally active site in squamous cell carcinoma (SQCC) tissue and cells. However, it is unclear whether RNA associated with cell migration exist at the DMP-1 gene locus in SQCC cells. We identified a novel promoter-associated non-coding RNA in the antisense strand of DMP-1 gene locus, promoter-associated non-coding RNA (panRNA)-DMP-1, by the RACE method in SQCC cells and tissues, and characterized the functions of panRNA-DMP-1 in EGF-driven SQCC cell migration. The inhibition of endogenous panRNA-DMP-1 expression by specific siRNAs and exogenous over-expression of panRNA-DMP-1 resulted in increased and suppressed cellular migration toward EGF in SQCC cells, respectively, and nuclear expression of panRNA-DMP-1 was induced by EGF stimulation. Mechanistically, suppression of panRNA-DMP-1 expression increased EGFR nuclear localization upon EGF treatment and nuclear panRNA-DMP-1 physically interacted with EGFR, which was confirmed by RNA immunoprecipitation assay using a bacteriophage-delivered PP7 RNA labeling system. Furthermore, co-immunoprecipitation assay revealed that suppression of panRNA-DMP-1 stabilized EGFR interaction with STAT3, a known co-transcription factors of EGFR, to induce migratory properties in many cancer cells. Based on these findings, panRNA-DMP-1 is an EGFR-associating RNA that inhibits the EGF-induced migratory properties of SQCC possibly by regulating EGFR nuclear localization and EGFR binding to STAT3.

Tomographic evaluation of direct pulp capping using a novel injectable treated dentin matrix hydrogel: a 2-year randomized controlled clinical trial.

OBJECTIVES: To assess clinically and radiographically the success of pulp capping procedure done in traumatically exposed permanent posterior teeth using a novel injectable treated dentin matrix hydrogel (TDMH), Biodentine, and MTA and to evaluate the formed dentin bridge under the capping materials using CBCT imaging. MATERIALS AND METHODS: 45 patients subjected to accidental traumatic pulp exposures by undergraduate dental students are allocated for this study. For each patient, a pulp capping procedure was done. TDMH was formed of TDM powder and sodium alginate to be injected and then hardened in the defect area. Patients were assigned to 3 groups: TDMH, Biodentine, and MTA, respectively, and returned to the clinic after 3, 6, 12, 18, and 24 months for clinical and radiographic examinations. Tomographic data, including thickness and density of formed dentin bridges, were evaluated at the end of the study period compared to the base line. Pulp sensitivity was evaluated throughout the study period using thermal testing and electric pulp tester. RESULTS: During the follow-up period, all patients were asymptomatic with no clinical signs and symptoms and revealed no radiographic signs of pathosis. However, tomographic evaluation showed the tested materials to have different levels of impact on formed dentin bridges with TDMH group resulted in significantly superior dentin bridges of a higher radiodensity and thickness than Biodentine and MTA. CONCLUSIONS: TDMH has a greater potential to induce dentin bridge formation than Biodentine and MTA under standardized conditions. Additionally, CBCT imaging was confirmed as a non-invasive and inclusive approach to evaluate the formed dentin bridges after pulp capping procedure. CLINICAL RELEVANCE: Direct pulp capping can be done successfully with this novel injectable pulp capping material in future clinical applications. TRIAL REGISTRATION: PACTR201901866476410.

Histological evaluation of the regenerative potential of a novel treated dentin matrix hydrogel in direct pulp capping.

OBJECTIVES: To produce a novel injectable treated dentin matrix hydrogel (TDMH) to be used as a novel pulp-capping agent for dentin regeneration compared with Biodentine and MTA. MATERIALS AND METHODS: Thirty intact fully erupted premolars scheduled to be extracted for orthodontic reasons were included. Pulps were mechanically exposed in the middle of the cavity floor. TDMH was composed of TDM powder (500-µm particle size) and sodium alginate as an injectable scaffold. The capped teeth were divided into three equal groups (n = 10): TDMH, Biodentine, and MTA respectively. Clinical examination and assessment of periapical response were performed. The teeth were extracted after 2-weeks and 2-month intervals, stained with hematoxylin-eosin, and categorized by using a histologic scoring system. Statistical analysis was performed using chi-square and Kruskal-Wallis test (p = 0.05). RESULTS: All teeth were vital during observation periods. Histological analysis after 2 months showed complete dentin bridge formation and absence of inflammatory pulp response with no significant differences between groups. However, the formed dentin was significantly thicker with the TDMH group with layers of well-arranged odontoblasts that were found to form a homogenous tubular structure with numerous dentinal tubule lines showing a positive trend to dentin regeneration. CONCLUSIONS: TDMH could achieve dentin regeneration and conservation of pulp vitality and might serve as a feasible natural substitute for silicate-based cements in restoring in vivo dentin defect in direct pulp-capping procedure. TRIAL REGISTRATION: PACTR201901866476410.