Claudia Brizuela - Academia.edu (original) (raw)

Papers by Claudia Brizuela

Research paper thumbnail of Human teeth with periapical pathosis after overinstrumentation and overfilling of the root canals: a scanning electron microscopic study

International Endodontic Journal, 1999

The aim of this study was to determine whether overinstrumentation followed by immediate overfill... more The aim of this study was to determine whether overinstrumentation followed by immediate overfilling could be a potential risk in the treatment of infected root canals. Thirty-five human teeth with infected root canals were overinstrumented and overfilled approximately 45 min after their extraction. The experimental teeth were enlarged up to size 40 and the overinstrumentation and overfilling were checked with the aid of a magnifying glass. The specimens were fixed in glutaraldehyde plus sodium cacodylate solution and prepared for scanning electron microscope examination. Bacteria were detected on the flute of the files and mostly at the root apices around the main foramen, remaining firmly attached to resorptive lacunae despite the fact that the apices had undergone great changes, including fracture or zipping. A control group consisting of 10 human teeth root canals containing vital pulps were also overinstrumented and overfilled. No bacteria were detected on the flutes of the files, at the apices or on the extruded master cone overfilling these samples. The high percentage of bacteria adhering to the resorptive lacunae or in the flutes of files used in overinstrumented human teeth with infected root canals carry a potential risk for postoperative pain, clinical discomfort and flare-ups. The hazards observed in these circumstances do not support the one-visit treatment of teeth having acute or chronic periapical abscesses.

Research paper thumbnail of Cell-Based Regenerative Endodontics for Treatment of Periapical Lesions: A Randomized, Controlled Phase I/II Clinical Trial

Journal of Dental Research, 2020

A randomized controlled phase I/II clinical trial was designed to evaluate the safety and efficac... more A randomized controlled phase I/II clinical trial was designed to evaluate the safety and efficacy of encapsulated human umbilical cord mesenchymal stem cells in a plasma-derived biomaterial for regenerative endodontic procedures (REPs) in mature permanent teeth with apical lesions. The trial included 36 patients with mature incisors, canines, or mandibular premolars showing pulp necrosis and apical periodontitis. Patients were randomly and equally allocated between experimental (REP) or conventional root canal treatment (ENDO) groups. On the first visit, cavity access and mechanical preparation of the root canal were performed. Calcium hydroxide medication was used, and the cavity was sealed. Three weeks later, patients were treated following their assigned protocol of ENDO or REP. Clinical follow-up examinations were performed at 6 and 12 mo. Categorical variables were evaluated by Fisher’s exact test. Quantitative variables were compared using the Mann-Whitney test. The evolution...

Research paper thumbnail of The Four Pillars for Successful Regenerative Therapy in Endodontics: Stem Cells, Biomaterials, Growth Factors, and Their Synergistic Interactions

Stem Cells International, Sep 19, 2022

Endodontics has made significant progress in regenerative approaches in recent years, thanks to a... more Endodontics has made significant progress in regenerative approaches in recent years, thanks to advances in biologically based procedures or regenerative endodontic therapy (RET). In recent years, our profession has witnessed a clear conceptual shift in this therapy. RET was initially based on a blood clot induced by apical bleeding without harvesting the patient's cells or cellfree RET. Later, the RET encompassed the three principles of tissue engineering, stromal/stem cells, scaffolds, and growth factors, aiming for the regeneration of a functional dentin pulp complex. The regenerated dental pulp will recover the protective mechanisms including innate immunity, tertiary dentin formation, and pain sensitivity. This comprehensive review covers the basic knowledge and practical information for translational applications of stem cell-based RET and tissue engineering procedures for the regeneration of dental pulp. It will also provide overall information on the emerging technologies in biological and synthetic matrices, biomaterials, and signaling molecules, recent advances in stem cell therapy, and updated experimental results. This review brings useful and timely clinical evidence for practitioners to understand the challenges faced for a successful cell-based RET and the importance of preserving or reestablishing tooth vitality. The clinical translation of these current bioengineering approaches will undoubtedly be beneficial to the future practice of endodontics.

Research paper thumbnail of Regenerative Endodontic Procedure in an Immature Permanent Incisor with Internal Root Resorption: a Case Report

Journal of Dentistry, Shiraz University of Medical Sciences, Jun 1, 2022

This report describes a regenerative endodontic procedure of an immature permanent incisor with i... more This report describes a regenerative endodontic procedure of an immature permanent incisor with internal root resorption (IRR) and 4-years follow-up. A healthy 8-year-old man was referred for treatment of tooth #9 after a traumatic intrusion. The periapical radiograph showed an IRR and an open apex with periradicular lesion. A diagnosis of pulp necrosis and chronic apical abscess was achieved. In the first appointment, under local anesthesia and rubber dam isolation, an access cavity was designed and the root canal was chemically cleaned under irrigation with 10 mL 1.5% sodium hypochlorite (NaOCl). The root canal was then dried and calcium hydroxide paste was placed. During the second appointment, the root canal was irrigated with 5 mL of 17% ethylenediaminetetraacetic acid (EDTA) for 5 minutes and dried. The blood clot was established in a time of 3 minutes after the bleeding from the periapical tissue was trigged. White mineral trioxide aggregate (MTA) was placed up to the amelocemental junction and the final restoration of the access cavity was carried out. During periodic clinical and radiographic follow-up, the patient remained symptom free, the periapical region was completely healed, inhibition of the root resorption process achieved, and formation of the new periodontal ligament as well as tooth widening development observed, meeting functional expectations after 48 months. The regenerative endodontic procedures are an available option to treat IRR in severely immature teeth. The available literature on the regenerative endodontic procedures applied to IRR treatment is limited, and more research is needed in this field.

Research paper thumbnail of Supplementary_Material - Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-derived Mesenchymal Stem Cells in Proliferation Rate, Migration Ability, and Angiogenic Potential

Supplementary_Material for Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-... more Supplementary_Material for Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-derived Mesenchymal Stem Cells in Proliferation Rate, Migration Ability, and Angiogenic Potential by Ioannis Angelopoulos, Claudia Brizuela, and Maroun Khoury in Cell Transplantation

Research paper thumbnail of Delivery of affordable and scalable encapsulated allogenic/autologous mesenchymal stem cells in coagulated platelet poor plasma for dental pulp regeneration

Scientific Reports, 2022

The main goal of regenerative endodontics procedures (REPs) is to revitalize teeth by the regener... more The main goal of regenerative endodontics procedures (REPs) is to revitalize teeth by the regeneration of healthy dental pulp. In this study, we evaluated the potential of combining a natural and accessible biomaterial based on Platelet Poor Plasma (PPP) as a support for dental pulp stem cells (DPSC) and umbilical cord mesenchymal stem cells (UC-MSC). A comparison study between the two cell sources revealed compatibility with the PPP based scaffold with differences noted in the proliferation and angiogenic properties in vitro. Additionally, the release of growth factors including VEGF, HGF and DMP-1, was detected in the media of cultured PPP and was enhanced by the presence of the encapsulated MSCs. Dentin-Discs from human molars were filled with PPP alone or with MSCs and implanted subcutaneously for 4 weeks in mice. Histological analysis of the MSC-PPP implants revealed a newly formed dentin-like structure evidenced by the expression of Dentin sialophosphoprotein (DSPP). Finally, ...

Research paper thumbnail of Allogeneic Cellular Therapy in a Mature Tooth with Apical Periodontitis and Accidental Root Perforation: A Case Report

Journal of Endodontics, 2020

Introduction: Cell therapy in regenerative endodontics introduces an alternative option to classi... more Introduction: Cell therapy in regenerative endodontics introduces an alternative option to classic treatment strategies for complex endodontic cases. The aim of this case report was to describe cell-based therapy using allogeneic umbilical cord mesenchymal stem cells (UC-MSCs) encapsulated in a bioscaffold for a complex case of a mature permanent tooth with apical periodontitis and accidental root perforation. Methods: A healthy 19-year-old man undergoing orthodontic treatment was referred for endodontic treatment in tooth #7; he was diagnosed with apical periodontitis during a previously initiated treatment associated with accidental perforation of the radicular cervical third. The root perforation was sealed with glass ionomer and composite resin, and the root canal was instrumented, disinfected, and dressed with calcium hydroxide. After 3 weeks, allogeneic UC-MSCs were encapsulated in plateletpoor plasma and then implanted into the root canal, and Biodentine (Septodont, Saint-Maurdes-Fosses, France) was placed below the cementoenamel junction. Finally, the tooth was restored with composite resin. Results: Follow-up examinations were performed 6 months and 1 year later. The examinations included periapical radiography, cone-beam computed tomographic imaging, and sensitivity and vitality tests. Radiographic and cone-beam computed tomographic images indicated remission of the apical lesion. Clinical evaluations revealed normal responses to percussion and palpation tests; the tooth was responsive to the electric pulp test, and the vitality test indicated low blood perfusion units. Conclusions: This case report reveals the potential use of allogeneic cellular therapy using encapsulated UC-MSCS in a platelet-poor plasma scaffold for a complex case of a permanent tooth with apical periodontitis and root perforation.

Research paper thumbnail of Personalized Cell Therapy for Pulpitis Using Autologous Dental Pulp Stem Cells and Leukocyte Platelet-rich Fibrin: A Case Report

Journal of Endodontics, 2019

Introduction: Regenerative endodontic procedures have emerged as a new treatment. The aim of this... more Introduction: Regenerative endodontic procedures have emerged as a new treatment. The aim of this case report was to describe a regenerative autologous cellular therapy using mesenchymal stem cells from inflamed dental pulp and leukocyte platelet-rich fibrin (L-PRF) in a mature tooth. Methods: A healthy 50year-old man consulting for spontaneous dental pain was referred for endodontic treatment in tooth #28, which was diagnosed with symptomatic irreversible pulpitis. Inflamed dental pulp was extracted and transported to a good manufacturing practice laboratory for the isolation and culture of dental pulp stem cells (DPSCs). L-PRF was obtained from the patient's blood and was introduced into the instrumented and disinfected root canal, and expanded DPSCs were inoculated into the clot. The cervical part of the root canal was sealed with Biodentine (Septodont, Saint-Maur-des-Fosses, France) and a composite resin. Results: Follow-up examinations were performed 6 months and 3 years later. The examinations included periapical radiographs (to measure the periapical index [PAI]), conebeam computed tomographic (CBCT) imaging, sensitivity, and vitality tests. Clinical evaluations revealed normal responses to percussion and palpation tests. The tooth had a delayed response to cold, and the electric pulp test was responsive. The PAI and CBCT imaging revealed that the periapical area remained normal with a PAI score of 1 and a CBCT PAI score of 0. The vitality test performed indicated low blood perfusion units. Conclusions: This case study reveals the potential use of a patient's own DPSCs and L-PRF as an alternative procedure for the treatment of pulpitis in mature permanent teeth. It also paves the way for the design of personalized cellbased clinical trials in regenerative endodontics.

Research paper thumbnail of Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-derived Mesenchymal Stem Cells in Proliferation Rate, Migration Ability, and Angiogenic Potential

Cell Transplantation, 2018

High donor variation makes comparison studies between different dental sources dubious. Dental ti... more High donor variation makes comparison studies between different dental sources dubious. Dental tissues offer a rare opportunity for comparing the biological characteristics of haploidentical mesenchymal stem cells (MSCs) isolated from the same donor. The objective was to identify the optimal dental source of MSCs through a biological and functional comparison of haploidentical MSCs from gingival (GMSCs) and dental pulp stem cells (DPSCs) focusing mainly on their angiogenic potential. The comparison study included (1) surface markers expression, (2) mesodermal differentiation capacity (chondrogenic, adipogenic, and osteogenic), (3) proliferation, (4) migration potential, (5) ability to form colony units, and (6) angiogenic potential in vitro and in vivo. Comparative analysis showed no difference in the immunophenotypic profile nor for the trilineage differentiation potential. Proliferation of GMSCs was higher than DPSCs at day 6 (2.6-fold higher, P < 0.05). GMSCs showed superior m...

Research paper thumbnail of MatrigelBD, A Biocompatible Scaffold that Improves Gingival Mesenchymal Stem Cells Proliferation

International Journal of Morphology, 2016

MatrigelBD is a hydrogel scaffold with three-dimensional intercrossed networks of hydrophilic pol... more MatrigelBD is a hydrogel scaffold with three-dimensional intercrossed networks of hydrophilic polymers with high water content. Human gingival tissue might represent a better source of MSCs, allowing these cells to be easily obtained in a relatively non-invasive way. The objective of this study was to evaluate the biocompatibility of MatrigelBD with GMSCs in vitro. Gingival connective tissue samples were obtained from healthy donors. Fresh tissue was minced and cultured during two weeks, after which cells at passage fourth were analyzed for their immune phenotype by flow cytometry. Differentiation into osteogenic, chondrogenic, and adipogenic lineages was induced and evaluated by culture staining. The "construct" was made of MatrigelBD with GMSC. To assess the biocompatibility, an MTT cellular proliferation assay was performed. The differentiation potential of the cells toward the osteogenic, adipogenic, and chondrogenic lineages was analyzed after 21 days of growth in MatrigelBD with induction differentiation media. The MTT analysis showed that MatrigelBD stimulated cell proliferation; the GMSCs maintained the expression of MSC markers. Importantly, the growth of GMSCs within the MatrigelBD did not interfere with the cell differentiation potential. These findings indicate that MatrigelBD is biocompatible with GMSCs, and this matrix improves cell proliferation in vitro.

Research paper thumbnail of BD-Matrigel with MSCg in a endodontic instrumented tooth

Objective: To assess biocompatibility of an hydrogel scaffold (BD-Matrigel) with MSCg in a tooth ... more Objective: To assess biocompatibility of an hydrogel scaffold (BD-Matrigel) with MSCg in a tooth model in vitro. Method: Mesenchymal stem cells were isolated from gingival connective tissue, immunophenotype was characterized by flow citometry, using specific antibodies markers. MSCg were differentiated into adipocytes, osteoblasts and chondroblasts following the criteria established by the ISCT. MSCg were cultured with BD-Matrigel, the construct was implanted in a tooth that was instrumented with Mtwo (VDW) rotary instrumentation system, then the tooth was cultured for 10 days. Biocompatibility was assessed by viability test, proliferation curve and cell type identification with histological cuts. Result: It was observed that the whole BD-Matrigel cultivated with MSCg in the tooth model, permitted proliferation, cell viability and cell function. It also was demonstrated the presence of cellular elements within the construct, subsequent to cultivation. Conclusion: BD-Matrigel proved ...

Research paper thumbnail of Regenerative endodontic procedure combined with non‐surgical root canal treatment in an immature permanent maxillary molar: A 7‐year follow‐up case report

Australian Endodontic Journal

Research paper thumbnail of An Innovative Regenerative Endodontic Procedure Using Leukocyte and Platelet-rich Fibrin Associated with Apical Surgery: A Case Report

Journal of endodontics, 2017

Regenerative endodontic procedures (REPs) associated with apical surgery could represent an alter... more Regenerative endodontic procedures (REPs) associated with apical surgery could represent an alternative treatment strategy for patients whose teeth present incomplete root formation and extensive apical lesions. Leukocyte platelet-rich fibrin (L-PRF) has potential benefits in REPs; it could promote apical root formation and optimal bone healing. The aim of this case report was to describe innovative regenerative endodontic therapy using L-PRF in the root canal and an extensive apical lesion in an immature tooth with dens invaginatus and asymptomatic apical periodontitis. A healthy 20-year-old woman was referred to the dental clinic of the Universidad de Los Andes, Santiago, Chile, for endodontic treatment in tooth # 22 with incomplete root development and an extensive apical lesion. The diagnosis was asymptomatic apical periodontitis associated with dens invaginatus type II. The patient was treated with an innovative approach using L-PRF in REPs associated with apical surgery. Follo...

Research paper thumbnail of Root and canal morphology of mandibular premolars using cone-beam computed tomography in a Chilean and Belgian subpopulation: a cross-sectional study

Oral Radiology, 2017

To describe mandibular premolar root and canal morphology and its variability in Chilean and Belg... more To describe mandibular premolar root and canal morphology and its variability in Chilean and Belgian samples using cone-beam computed tomography. A cross-sectional and descriptive study was conducted. A total of 402 mandibular premolars were examined using cone-beam computed tomography images of Chilean and Belgian patients. Premolars that met the inclusion criteria were studied in relation to the number of roots, number of canals, root canal configuration, presence of C-shaped configuration, tooth length, and root length. The mandibular first premolar frequently presented with one root (94% Chilean; 100% Belgian), as did the mandibular second premolar (99% Chilean; 98% Belgian). One canal was present in 69% of Chilean, and 83% of Belgian mandibular first premolars, and in 95% of Chilean and 91% of Belgian second premolars. A type I root canal configuration was found in 69% of Chilean and 83% of Belgian first premolars and in 95% of Chilean and 92% of Belgian second premolars. The anatomical parameters analyzed in the Chilean and Belgian samples are similar. However, it should be noted that there are anatomical variations in mandibular premolars.

Research paper thumbnail of Direct Pulp Capping with Calcium Hydroxide, Mineral Trioxide Aggregate, and Biodentine in Permanent Young Teeth with Caries: A Randomized Clinical Trial

Journal of Endodontics, 2017

Introduction: Direct pulp capping treatment is intended to preserve pulp vitality, to avoid or re... more Introduction: Direct pulp capping treatment is intended to preserve pulp vitality, to avoid or retard root canal treatment, and, in cases with an open apex, to allow continued root development. Historically, calcium hydroxide (CH) was the gold standard material, but nowadays calcium silicate materials (CSMs) are displacing CH because of their high bioactivity, biocompatibility, sealing ability, and mechanical properties. However, more randomized clinical trials are needed to confirm the appropriateness of CSMs as replacement materials for CH in direct pulp capping procedures. Methods: A randomized clinical trial was conducted that included 169 patients (mean age, 11.3 years) from the Maipo district (Chile). The inclusion criterion was patients with 1 carious permanent tooth with pulpal exposure, a candidate for a direct pulp capping procedure. The patients were randomly allocated to one of the experimental groups (CH, Biodentine, or mineral trioxide aggregate [MTA]). Clinical follow-up examinations were performed at 1 week, 3 months, 6 months, and 1 year. The Fisher exact test was performed. Results: At the follow-up examination at 1 week, the patients showed 100% clinical success. At 3 months, there was 1 failure in the CH group. At 6 months, there were 4 new failures (1 in the CH group and 3 in the MTA group). At 1 year, there was another failure in the CH group. There were no statistically significant differences among the experimental groups. Conclusions: CSMs appear to be suitable materials to replace CH. Although no significant differences were found among the materials studied, Biodentine and MTA offered some advantages over CH.

Research paper thumbnail of Characterization of mandibular molar root and canal morphology using cone beam computed tomography and its variability in Belgian and Chilean population samples

Imaging Science in Dentistry, 2015

Purpose: This study used cone-beam computed tomography (CBCT) to characterize mandibular molar ro... more Purpose: This study used cone-beam computed tomography (CBCT) to characterize mandibular molar root and canal morphology and its variability in Belgian and Chilean population samples. Materials and Methods: We analyzed the CBCT images of 515 mandibular molars (257 from Belgium and 258 from Chile). Molars meeting the inclusion criteria were analyzed to determine (1) the number of roots; (2) the root canal configuration; (3) the presence of a curved canal in the cross-sectional image of the distal root in the mandibular first molar and (4) the presence of a C-shaped canal in the second mandibular molar. A descriptive analysis was performed. The association between national origin and the presence of a curved or C-shaped canal was evaluated using the chi-squared test. results: The most common configurations in the mesial root of both molars were type V and type III. In the distal root, type I canal configuration was the most common. Curvature in the cross-sectional image was found in 25% of the distal canals of the mandibular first molars in the Belgian population, compared to 11% in the Chilean population. The prevalence of C-shaped canals was 10% or less in both populations. Conclusion: In cases of unclear or complex root and canal morphology in the mandibular molars, CBCT imaging might assist endodontic specialists in making an accurate diagnosis and in treatment planning. (Imaging Sci Dent 2015; 45: 95-101

Research paper thumbnail of Mesenchymal Stem Cells Derived from Human Inflamed Dental Pulp Exhibit Impaired Immunomodulatory Capacity In Vitro

Journal of Endodontics, 2020

Abstract Introduction Dental pulp stem cells (DPSC) are very attractive in regenerative medicine.... more Abstract Introduction Dental pulp stem cells (DPSC) are very attractive in regenerative medicine. In this study, we focused on the characterization of the functional properties of mesenchymal stem cells derived from DPSCs. Currently, it is unknown whether inflammatory conditions present in an inflamed dental pulp tissue could alter the immunomodulatory properties of DPSCs. This study aimed to evaluate the immunomodulatory capacity in vitro of DPSCs derived from healthy and inflamed dental pulp. Methods DPSCs from 10 healthy and inflamed dental pulps (irreversible pulpitis) were characterized according to the minimal criteria of the International Society for Cell Therapy, proliferation, differential potential, and colony-forming units. Furthermore, the immunomodulatory capacity of DPSCs was tested on the proliferation of T lymphocytes by flow cytometry and the in vitro enzyme activity of indoleamine 2, 3-dioxygenase. Results There were no significant differences in the DPSC characteristics and properties such as immunophenotype, tridifferentiation, colony-forming units, and proliferation of the DPSCs derived from normal and inflamed pulp tissue. Furthermore, there were significant differences in the immunomodulatory capacity of DPSCs obtained from human healthy dental pulp and with the diagnosis of irreversible pulpitis. Conclusions Our results showed that DPSCs isolated from inflamed dental pulp showed typical characteristics of MSCs and diminished immunosuppressive capacity in vitro in comparison with MSCs derived from healthy dental pulp. Further investigation in vivo is needed to clarify the mechanism of this diminished immunosuppressive capacity.

Research paper thumbnail of Inflammatory biomarkers in dentinal fluid as an approach to molecular diagnostics in pulpitis

International Endodontic Journal, 2020

AIM To explore a set of inflammatory biomarkers obtained from dentinal fluid (DF) from patients w... more AIM To explore a set of inflammatory biomarkers obtained from dentinal fluid (DF) from patients with symptomatic irreversible pulpitis (IP), reversible pulpitis (RP) and normal pulp (NP). METHODOLOGY A cross-sectional exploratory study was performed, recruiting 64 patients on the basis of their respective pulp condition. DF samples were obtained from all patients (23, from IP patients; 20, from RP patients; and 21, from NP patients). Quantification of biomarkers was performed using a Luminex® MAGPIX platform system and multiplex assay kits. The Kruskal Wallis test was used for comparisons with regard to pulp state. A simple logistic regression model and the odds ratio (OR) with a 95% level of confidence (p=0.05) were used to evaluate associations between biomarker levels and pulpal diagnosis. The performance discrimination of the biomarkers was evaluated through the construction of a receiver operating characteristic (ROC) curve by calculating the area under the curve (AUC) for IP versus RP after logistic regression modeling. Youden criteria were used to establish cutoff points for biomarkers alone with AUC>70 and p-value<0.05, or estimated probabilities from the multivariable logistic model. RESULTS The biomarkers that had significantly higher values in participants with IP versus RP were IL-1α, VEGF-α and FGF-acid (p<0.05). FGF-acid (OR: 12.62; p = 0.0085; CI 95% 1.91-83.29) and VEGF-α (OR: 2.61; p = 0.0252; CI 95% 1.13-6.03) were associated with pulp diagnoses of IP versus RP. The AUC-ROC curve for FGF-acid was 0.79. The model containing FGF-acid, IL-1α, IL-6, and TIMP-1 had an AUC-ROC of 0.92 for IP versus RP with a significant difference from the FGF-acid ROC curve (p = 0.0231). CONCLUSIONS Dentinal fluid could be used to assay pulpal mediators in the molecular diagnosis of pulpitis. Despite the limitation of the clinical diagnostics used the present study, it was possible to detect a difference between irreversible symptomatic pulpitis and reversible pulpitis associated with the following combined biomarkers: FGF-acid + IL-6 + IL-1α, + TIMP-1.

Research paper thumbnail of Translational pathway of scalable, allogenic encapsulated Mesenchymal Stem Cells for Dental Pulp Regeneration. RanoKure a controlled Phase I/II clinical trial designed to evaluate the survival of mature permanent teeth with apical lesion following a regenerative endodontics procedure

Research paper thumbnail of Histological transformations of the dental pulp as possible indicator of post mortem interval: a pilot study

Forensic Science International

The correct estimation of the post mortem interval (PMI) can be crucial on the success of a foren... more The correct estimation of the post mortem interval (PMI) can be crucial on the success of a forensic investigation. Diverse methods have been used to estimate PMI, considering physical changes that occur after death, such as mortis algor, livor mortis, among others. Degradation after death of dental pulp is a complex process that has not yet been studied thoroughly. It has been described that pulp RNA degradation could be an indicator of PMI, however this study is limited to 6 days. The tooth is the hardest organ of the human body, and within is confined dental pulp. The pulp morphology is defined as a lax conjunctive tissue with great sensory innervation, abundant microcirculation and great presence of groups of cell types. The aim of this study is to describe the potential use of pulp post mortem alterations to estimate PMI, using a new methodology that will allow obtainment of pulp tissue to be used for histomorphological analysis. The current study will identify potential histological indicators in dental pulp tissue to estimate PMI in time intervals of 24h, 1 month, 3 months and 6 months. This study used 26 teeth from individuals with known PMI of 24h, 1 month, 3 months or 6 months. All samples were manipulated with the new methodology (Carrasco, P. and Inostroza C. inventors; Universidad de los Andes, assignee. Forensic identification, post mortem interval estimation and cause of death determination by recovery of dental tissue. United State patent US 61/826,558 23.05.2013) to extract pulp tissue without the destruction of the tooth. The dental pulp tissues obtained were fixed in formalin for the subsequent generation of histological sections, stained with Hematoxylin Eosin and Masson&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s Trichrome. All sections were observed under an optical microscope using magnifications of 10× and 40×. The microscopic analysis of the samples showed a progressive transformation of the cellular components and fibers of dental pulp along PMI. These results allowed creating a chart of qualitative and quantitative parameters to be used on the estimation on PMI based on microscopic degradation of dental pulp. The histological transformations of dental pulp as a function of time can be used as PMI indicators.

Research paper thumbnail of Human teeth with periapical pathosis after overinstrumentation and overfilling of the root canals: a scanning electron microscopic study

International Endodontic Journal, 1999

The aim of this study was to determine whether overinstrumentation followed by immediate overfill... more The aim of this study was to determine whether overinstrumentation followed by immediate overfilling could be a potential risk in the treatment of infected root canals. Thirty-five human teeth with infected root canals were overinstrumented and overfilled approximately 45 min after their extraction. The experimental teeth were enlarged up to size 40 and the overinstrumentation and overfilling were checked with the aid of a magnifying glass. The specimens were fixed in glutaraldehyde plus sodium cacodylate solution and prepared for scanning electron microscope examination. Bacteria were detected on the flute of the files and mostly at the root apices around the main foramen, remaining firmly attached to resorptive lacunae despite the fact that the apices had undergone great changes, including fracture or zipping. A control group consisting of 10 human teeth root canals containing vital pulps were also overinstrumented and overfilled. No bacteria were detected on the flutes of the files, at the apices or on the extruded master cone overfilling these samples. The high percentage of bacteria adhering to the resorptive lacunae or in the flutes of files used in overinstrumented human teeth with infected root canals carry a potential risk for postoperative pain, clinical discomfort and flare-ups. The hazards observed in these circumstances do not support the one-visit treatment of teeth having acute or chronic periapical abscesses.

Research paper thumbnail of Cell-Based Regenerative Endodontics for Treatment of Periapical Lesions: A Randomized, Controlled Phase I/II Clinical Trial

Journal of Dental Research, 2020

A randomized controlled phase I/II clinical trial was designed to evaluate the safety and efficac... more A randomized controlled phase I/II clinical trial was designed to evaluate the safety and efficacy of encapsulated human umbilical cord mesenchymal stem cells in a plasma-derived biomaterial for regenerative endodontic procedures (REPs) in mature permanent teeth with apical lesions. The trial included 36 patients with mature incisors, canines, or mandibular premolars showing pulp necrosis and apical periodontitis. Patients were randomly and equally allocated between experimental (REP) or conventional root canal treatment (ENDO) groups. On the first visit, cavity access and mechanical preparation of the root canal were performed. Calcium hydroxide medication was used, and the cavity was sealed. Three weeks later, patients were treated following their assigned protocol of ENDO or REP. Clinical follow-up examinations were performed at 6 and 12 mo. Categorical variables were evaluated by Fisher’s exact test. Quantitative variables were compared using the Mann-Whitney test. The evolution...

Research paper thumbnail of The Four Pillars for Successful Regenerative Therapy in Endodontics: Stem Cells, Biomaterials, Growth Factors, and Their Synergistic Interactions

Stem Cells International, Sep 19, 2022

Endodontics has made significant progress in regenerative approaches in recent years, thanks to a... more Endodontics has made significant progress in regenerative approaches in recent years, thanks to advances in biologically based procedures or regenerative endodontic therapy (RET). In recent years, our profession has witnessed a clear conceptual shift in this therapy. RET was initially based on a blood clot induced by apical bleeding without harvesting the patient's cells or cellfree RET. Later, the RET encompassed the three principles of tissue engineering, stromal/stem cells, scaffolds, and growth factors, aiming for the regeneration of a functional dentin pulp complex. The regenerated dental pulp will recover the protective mechanisms including innate immunity, tertiary dentin formation, and pain sensitivity. This comprehensive review covers the basic knowledge and practical information for translational applications of stem cell-based RET and tissue engineering procedures for the regeneration of dental pulp. It will also provide overall information on the emerging technologies in biological and synthetic matrices, biomaterials, and signaling molecules, recent advances in stem cell therapy, and updated experimental results. This review brings useful and timely clinical evidence for practitioners to understand the challenges faced for a successful cell-based RET and the importance of preserving or reestablishing tooth vitality. The clinical translation of these current bioengineering approaches will undoubtedly be beneficial to the future practice of endodontics.

Research paper thumbnail of Regenerative Endodontic Procedure in an Immature Permanent Incisor with Internal Root Resorption: a Case Report

Journal of Dentistry, Shiraz University of Medical Sciences, Jun 1, 2022

This report describes a regenerative endodontic procedure of an immature permanent incisor with i... more This report describes a regenerative endodontic procedure of an immature permanent incisor with internal root resorption (IRR) and 4-years follow-up. A healthy 8-year-old man was referred for treatment of tooth #9 after a traumatic intrusion. The periapical radiograph showed an IRR and an open apex with periradicular lesion. A diagnosis of pulp necrosis and chronic apical abscess was achieved. In the first appointment, under local anesthesia and rubber dam isolation, an access cavity was designed and the root canal was chemically cleaned under irrigation with 10 mL 1.5% sodium hypochlorite (NaOCl). The root canal was then dried and calcium hydroxide paste was placed. During the second appointment, the root canal was irrigated with 5 mL of 17% ethylenediaminetetraacetic acid (EDTA) for 5 minutes and dried. The blood clot was established in a time of 3 minutes after the bleeding from the periapical tissue was trigged. White mineral trioxide aggregate (MTA) was placed up to the amelocemental junction and the final restoration of the access cavity was carried out. During periodic clinical and radiographic follow-up, the patient remained symptom free, the periapical region was completely healed, inhibition of the root resorption process achieved, and formation of the new periodontal ligament as well as tooth widening development observed, meeting functional expectations after 48 months. The regenerative endodontic procedures are an available option to treat IRR in severely immature teeth. The available literature on the regenerative endodontic procedures applied to IRR treatment is limited, and more research is needed in this field.

Research paper thumbnail of Supplementary_Material - Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-derived Mesenchymal Stem Cells in Proliferation Rate, Migration Ability, and Angiogenic Potential

Supplementary_Material for Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-... more Supplementary_Material for Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-derived Mesenchymal Stem Cells in Proliferation Rate, Migration Ability, and Angiogenic Potential by Ioannis Angelopoulos, Claudia Brizuela, and Maroun Khoury in Cell Transplantation

Research paper thumbnail of Delivery of affordable and scalable encapsulated allogenic/autologous mesenchymal stem cells in coagulated platelet poor plasma for dental pulp regeneration

Scientific Reports, 2022

The main goal of regenerative endodontics procedures (REPs) is to revitalize teeth by the regener... more The main goal of regenerative endodontics procedures (REPs) is to revitalize teeth by the regeneration of healthy dental pulp. In this study, we evaluated the potential of combining a natural and accessible biomaterial based on Platelet Poor Plasma (PPP) as a support for dental pulp stem cells (DPSC) and umbilical cord mesenchymal stem cells (UC-MSC). A comparison study between the two cell sources revealed compatibility with the PPP based scaffold with differences noted in the proliferation and angiogenic properties in vitro. Additionally, the release of growth factors including VEGF, HGF and DMP-1, was detected in the media of cultured PPP and was enhanced by the presence of the encapsulated MSCs. Dentin-Discs from human molars were filled with PPP alone or with MSCs and implanted subcutaneously for 4 weeks in mice. Histological analysis of the MSC-PPP implants revealed a newly formed dentin-like structure evidenced by the expression of Dentin sialophosphoprotein (DSPP). Finally, ...

Research paper thumbnail of Allogeneic Cellular Therapy in a Mature Tooth with Apical Periodontitis and Accidental Root Perforation: A Case Report

Journal of Endodontics, 2020

Introduction: Cell therapy in regenerative endodontics introduces an alternative option to classi... more Introduction: Cell therapy in regenerative endodontics introduces an alternative option to classic treatment strategies for complex endodontic cases. The aim of this case report was to describe cell-based therapy using allogeneic umbilical cord mesenchymal stem cells (UC-MSCs) encapsulated in a bioscaffold for a complex case of a mature permanent tooth with apical periodontitis and accidental root perforation. Methods: A healthy 19-year-old man undergoing orthodontic treatment was referred for endodontic treatment in tooth #7; he was diagnosed with apical periodontitis during a previously initiated treatment associated with accidental perforation of the radicular cervical third. The root perforation was sealed with glass ionomer and composite resin, and the root canal was instrumented, disinfected, and dressed with calcium hydroxide. After 3 weeks, allogeneic UC-MSCs were encapsulated in plateletpoor plasma and then implanted into the root canal, and Biodentine (Septodont, Saint-Maurdes-Fosses, France) was placed below the cementoenamel junction. Finally, the tooth was restored with composite resin. Results: Follow-up examinations were performed 6 months and 1 year later. The examinations included periapical radiography, cone-beam computed tomographic imaging, and sensitivity and vitality tests. Radiographic and cone-beam computed tomographic images indicated remission of the apical lesion. Clinical evaluations revealed normal responses to percussion and palpation tests; the tooth was responsive to the electric pulp test, and the vitality test indicated low blood perfusion units. Conclusions: This case report reveals the potential use of allogeneic cellular therapy using encapsulated UC-MSCS in a platelet-poor plasma scaffold for a complex case of a permanent tooth with apical periodontitis and root perforation.

Research paper thumbnail of Personalized Cell Therapy for Pulpitis Using Autologous Dental Pulp Stem Cells and Leukocyte Platelet-rich Fibrin: A Case Report

Journal of Endodontics, 2019

Introduction: Regenerative endodontic procedures have emerged as a new treatment. The aim of this... more Introduction: Regenerative endodontic procedures have emerged as a new treatment. The aim of this case report was to describe a regenerative autologous cellular therapy using mesenchymal stem cells from inflamed dental pulp and leukocyte platelet-rich fibrin (L-PRF) in a mature tooth. Methods: A healthy 50year-old man consulting for spontaneous dental pain was referred for endodontic treatment in tooth #28, which was diagnosed with symptomatic irreversible pulpitis. Inflamed dental pulp was extracted and transported to a good manufacturing practice laboratory for the isolation and culture of dental pulp stem cells (DPSCs). L-PRF was obtained from the patient's blood and was introduced into the instrumented and disinfected root canal, and expanded DPSCs were inoculated into the clot. The cervical part of the root canal was sealed with Biodentine (Septodont, Saint-Maur-des-Fosses, France) and a composite resin. Results: Follow-up examinations were performed 6 months and 3 years later. The examinations included periapical radiographs (to measure the periapical index [PAI]), conebeam computed tomographic (CBCT) imaging, sensitivity, and vitality tests. Clinical evaluations revealed normal responses to percussion and palpation tests. The tooth had a delayed response to cold, and the electric pulp test was responsive. The PAI and CBCT imaging revealed that the periapical area remained normal with a PAI score of 1 and a CBCT PAI score of 0. The vitality test performed indicated low blood perfusion units. Conclusions: This case study reveals the potential use of a patient's own DPSCs and L-PRF as an alternative procedure for the treatment of pulpitis in mature permanent teeth. It also paves the way for the design of personalized cellbased clinical trials in regenerative endodontics.

Research paper thumbnail of Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-derived Mesenchymal Stem Cells in Proliferation Rate, Migration Ability, and Angiogenic Potential

Cell Transplantation, 2018

High donor variation makes comparison studies between different dental sources dubious. Dental ti... more High donor variation makes comparison studies between different dental sources dubious. Dental tissues offer a rare opportunity for comparing the biological characteristics of haploidentical mesenchymal stem cells (MSCs) isolated from the same donor. The objective was to identify the optimal dental source of MSCs through a biological and functional comparison of haploidentical MSCs from gingival (GMSCs) and dental pulp stem cells (DPSCs) focusing mainly on their angiogenic potential. The comparison study included (1) surface markers expression, (2) mesodermal differentiation capacity (chondrogenic, adipogenic, and osteogenic), (3) proliferation, (4) migration potential, (5) ability to form colony units, and (6) angiogenic potential in vitro and in vivo. Comparative analysis showed no difference in the immunophenotypic profile nor for the trilineage differentiation potential. Proliferation of GMSCs was higher than DPSCs at day 6 (2.6-fold higher, P < 0.05). GMSCs showed superior m...

Research paper thumbnail of MatrigelBD, A Biocompatible Scaffold that Improves Gingival Mesenchymal Stem Cells Proliferation

International Journal of Morphology, 2016

MatrigelBD is a hydrogel scaffold with three-dimensional intercrossed networks of hydrophilic pol... more MatrigelBD is a hydrogel scaffold with three-dimensional intercrossed networks of hydrophilic polymers with high water content. Human gingival tissue might represent a better source of MSCs, allowing these cells to be easily obtained in a relatively non-invasive way. The objective of this study was to evaluate the biocompatibility of MatrigelBD with GMSCs in vitro. Gingival connective tissue samples were obtained from healthy donors. Fresh tissue was minced and cultured during two weeks, after which cells at passage fourth were analyzed for their immune phenotype by flow cytometry. Differentiation into osteogenic, chondrogenic, and adipogenic lineages was induced and evaluated by culture staining. The "construct" was made of MatrigelBD with GMSC. To assess the biocompatibility, an MTT cellular proliferation assay was performed. The differentiation potential of the cells toward the osteogenic, adipogenic, and chondrogenic lineages was analyzed after 21 days of growth in MatrigelBD with induction differentiation media. The MTT analysis showed that MatrigelBD stimulated cell proliferation; the GMSCs maintained the expression of MSC markers. Importantly, the growth of GMSCs within the MatrigelBD did not interfere with the cell differentiation potential. These findings indicate that MatrigelBD is biocompatible with GMSCs, and this matrix improves cell proliferation in vitro.

Research paper thumbnail of BD-Matrigel with MSCg in a endodontic instrumented tooth

Objective: To assess biocompatibility of an hydrogel scaffold (BD-Matrigel) with MSCg in a tooth ... more Objective: To assess biocompatibility of an hydrogel scaffold (BD-Matrigel) with MSCg in a tooth model in vitro. Method: Mesenchymal stem cells were isolated from gingival connective tissue, immunophenotype was characterized by flow citometry, using specific antibodies markers. MSCg were differentiated into adipocytes, osteoblasts and chondroblasts following the criteria established by the ISCT. MSCg were cultured with BD-Matrigel, the construct was implanted in a tooth that was instrumented with Mtwo (VDW) rotary instrumentation system, then the tooth was cultured for 10 days. Biocompatibility was assessed by viability test, proliferation curve and cell type identification with histological cuts. Result: It was observed that the whole BD-Matrigel cultivated with MSCg in the tooth model, permitted proliferation, cell viability and cell function. It also was demonstrated the presence of cellular elements within the construct, subsequent to cultivation. Conclusion: BD-Matrigel proved ...

Research paper thumbnail of Regenerative endodontic procedure combined with non‐surgical root canal treatment in an immature permanent maxillary molar: A 7‐year follow‐up case report

Australian Endodontic Journal

Research paper thumbnail of An Innovative Regenerative Endodontic Procedure Using Leukocyte and Platelet-rich Fibrin Associated with Apical Surgery: A Case Report

Journal of endodontics, 2017

Regenerative endodontic procedures (REPs) associated with apical surgery could represent an alter... more Regenerative endodontic procedures (REPs) associated with apical surgery could represent an alternative treatment strategy for patients whose teeth present incomplete root formation and extensive apical lesions. Leukocyte platelet-rich fibrin (L-PRF) has potential benefits in REPs; it could promote apical root formation and optimal bone healing. The aim of this case report was to describe innovative regenerative endodontic therapy using L-PRF in the root canal and an extensive apical lesion in an immature tooth with dens invaginatus and asymptomatic apical periodontitis. A healthy 20-year-old woman was referred to the dental clinic of the Universidad de Los Andes, Santiago, Chile, for endodontic treatment in tooth # 22 with incomplete root development and an extensive apical lesion. The diagnosis was asymptomatic apical periodontitis associated with dens invaginatus type II. The patient was treated with an innovative approach using L-PRF in REPs associated with apical surgery. Follo...

Research paper thumbnail of Root and canal morphology of mandibular premolars using cone-beam computed tomography in a Chilean and Belgian subpopulation: a cross-sectional study

Oral Radiology, 2017

To describe mandibular premolar root and canal morphology and its variability in Chilean and Belg... more To describe mandibular premolar root and canal morphology and its variability in Chilean and Belgian samples using cone-beam computed tomography. A cross-sectional and descriptive study was conducted. A total of 402 mandibular premolars were examined using cone-beam computed tomography images of Chilean and Belgian patients. Premolars that met the inclusion criteria were studied in relation to the number of roots, number of canals, root canal configuration, presence of C-shaped configuration, tooth length, and root length. The mandibular first premolar frequently presented with one root (94% Chilean; 100% Belgian), as did the mandibular second premolar (99% Chilean; 98% Belgian). One canal was present in 69% of Chilean, and 83% of Belgian mandibular first premolars, and in 95% of Chilean and 91% of Belgian second premolars. A type I root canal configuration was found in 69% of Chilean and 83% of Belgian first premolars and in 95% of Chilean and 92% of Belgian second premolars. The anatomical parameters analyzed in the Chilean and Belgian samples are similar. However, it should be noted that there are anatomical variations in mandibular premolars.

Research paper thumbnail of Direct Pulp Capping with Calcium Hydroxide, Mineral Trioxide Aggregate, and Biodentine in Permanent Young Teeth with Caries: A Randomized Clinical Trial

Journal of Endodontics, 2017

Introduction: Direct pulp capping treatment is intended to preserve pulp vitality, to avoid or re... more Introduction: Direct pulp capping treatment is intended to preserve pulp vitality, to avoid or retard root canal treatment, and, in cases with an open apex, to allow continued root development. Historically, calcium hydroxide (CH) was the gold standard material, but nowadays calcium silicate materials (CSMs) are displacing CH because of their high bioactivity, biocompatibility, sealing ability, and mechanical properties. However, more randomized clinical trials are needed to confirm the appropriateness of CSMs as replacement materials for CH in direct pulp capping procedures. Methods: A randomized clinical trial was conducted that included 169 patients (mean age, 11.3 years) from the Maipo district (Chile). The inclusion criterion was patients with 1 carious permanent tooth with pulpal exposure, a candidate for a direct pulp capping procedure. The patients were randomly allocated to one of the experimental groups (CH, Biodentine, or mineral trioxide aggregate [MTA]). Clinical follow-up examinations were performed at 1 week, 3 months, 6 months, and 1 year. The Fisher exact test was performed. Results: At the follow-up examination at 1 week, the patients showed 100% clinical success. At 3 months, there was 1 failure in the CH group. At 6 months, there were 4 new failures (1 in the CH group and 3 in the MTA group). At 1 year, there was another failure in the CH group. There were no statistically significant differences among the experimental groups. Conclusions: CSMs appear to be suitable materials to replace CH. Although no significant differences were found among the materials studied, Biodentine and MTA offered some advantages over CH.

Research paper thumbnail of Characterization of mandibular molar root and canal morphology using cone beam computed tomography and its variability in Belgian and Chilean population samples

Imaging Science in Dentistry, 2015

Purpose: This study used cone-beam computed tomography (CBCT) to characterize mandibular molar ro... more Purpose: This study used cone-beam computed tomography (CBCT) to characterize mandibular molar root and canal morphology and its variability in Belgian and Chilean population samples. Materials and Methods: We analyzed the CBCT images of 515 mandibular molars (257 from Belgium and 258 from Chile). Molars meeting the inclusion criteria were analyzed to determine (1) the number of roots; (2) the root canal configuration; (3) the presence of a curved canal in the cross-sectional image of the distal root in the mandibular first molar and (4) the presence of a C-shaped canal in the second mandibular molar. A descriptive analysis was performed. The association between national origin and the presence of a curved or C-shaped canal was evaluated using the chi-squared test. results: The most common configurations in the mesial root of both molars were type V and type III. In the distal root, type I canal configuration was the most common. Curvature in the cross-sectional image was found in 25% of the distal canals of the mandibular first molars in the Belgian population, compared to 11% in the Chilean population. The prevalence of C-shaped canals was 10% or less in both populations. Conclusion: In cases of unclear or complex root and canal morphology in the mandibular molars, CBCT imaging might assist endodontic specialists in making an accurate diagnosis and in treatment planning. (Imaging Sci Dent 2015; 45: 95-101

Research paper thumbnail of Mesenchymal Stem Cells Derived from Human Inflamed Dental Pulp Exhibit Impaired Immunomodulatory Capacity In Vitro

Journal of Endodontics, 2020

Abstract Introduction Dental pulp stem cells (DPSC) are very attractive in regenerative medicine.... more Abstract Introduction Dental pulp stem cells (DPSC) are very attractive in regenerative medicine. In this study, we focused on the characterization of the functional properties of mesenchymal stem cells derived from DPSCs. Currently, it is unknown whether inflammatory conditions present in an inflamed dental pulp tissue could alter the immunomodulatory properties of DPSCs. This study aimed to evaluate the immunomodulatory capacity in vitro of DPSCs derived from healthy and inflamed dental pulp. Methods DPSCs from 10 healthy and inflamed dental pulps (irreversible pulpitis) were characterized according to the minimal criteria of the International Society for Cell Therapy, proliferation, differential potential, and colony-forming units. Furthermore, the immunomodulatory capacity of DPSCs was tested on the proliferation of T lymphocytes by flow cytometry and the in vitro enzyme activity of indoleamine 2, 3-dioxygenase. Results There were no significant differences in the DPSC characteristics and properties such as immunophenotype, tridifferentiation, colony-forming units, and proliferation of the DPSCs derived from normal and inflamed pulp tissue. Furthermore, there were significant differences in the immunomodulatory capacity of DPSCs obtained from human healthy dental pulp and with the diagnosis of irreversible pulpitis. Conclusions Our results showed that DPSCs isolated from inflamed dental pulp showed typical characteristics of MSCs and diminished immunosuppressive capacity in vitro in comparison with MSCs derived from healthy dental pulp. Further investigation in vivo is needed to clarify the mechanism of this diminished immunosuppressive capacity.

Research paper thumbnail of Inflammatory biomarkers in dentinal fluid as an approach to molecular diagnostics in pulpitis

International Endodontic Journal, 2020

AIM To explore a set of inflammatory biomarkers obtained from dentinal fluid (DF) from patients w... more AIM To explore a set of inflammatory biomarkers obtained from dentinal fluid (DF) from patients with symptomatic irreversible pulpitis (IP), reversible pulpitis (RP) and normal pulp (NP). METHODOLOGY A cross-sectional exploratory study was performed, recruiting 64 patients on the basis of their respective pulp condition. DF samples were obtained from all patients (23, from IP patients; 20, from RP patients; and 21, from NP patients). Quantification of biomarkers was performed using a Luminex® MAGPIX platform system and multiplex assay kits. The Kruskal Wallis test was used for comparisons with regard to pulp state. A simple logistic regression model and the odds ratio (OR) with a 95% level of confidence (p=0.05) were used to evaluate associations between biomarker levels and pulpal diagnosis. The performance discrimination of the biomarkers was evaluated through the construction of a receiver operating characteristic (ROC) curve by calculating the area under the curve (AUC) for IP versus RP after logistic regression modeling. Youden criteria were used to establish cutoff points for biomarkers alone with AUC>70 and p-value<0.05, or estimated probabilities from the multivariable logistic model. RESULTS The biomarkers that had significantly higher values in participants with IP versus RP were IL-1α, VEGF-α and FGF-acid (p<0.05). FGF-acid (OR: 12.62; p = 0.0085; CI 95% 1.91-83.29) and VEGF-α (OR: 2.61; p = 0.0252; CI 95% 1.13-6.03) were associated with pulp diagnoses of IP versus RP. The AUC-ROC curve for FGF-acid was 0.79. The model containing FGF-acid, IL-1α, IL-6, and TIMP-1 had an AUC-ROC of 0.92 for IP versus RP with a significant difference from the FGF-acid ROC curve (p = 0.0231). CONCLUSIONS Dentinal fluid could be used to assay pulpal mediators in the molecular diagnosis of pulpitis. Despite the limitation of the clinical diagnostics used the present study, it was possible to detect a difference between irreversible symptomatic pulpitis and reversible pulpitis associated with the following combined biomarkers: FGF-acid + IL-6 + IL-1α, + TIMP-1.

Research paper thumbnail of Translational pathway of scalable, allogenic encapsulated Mesenchymal Stem Cells for Dental Pulp Regeneration. RanoKure a controlled Phase I/II clinical trial designed to evaluate the survival of mature permanent teeth with apical lesion following a regenerative endodontics procedure

Research paper thumbnail of Histological transformations of the dental pulp as possible indicator of post mortem interval: a pilot study

Forensic Science International

The correct estimation of the post mortem interval (PMI) can be crucial on the success of a foren... more The correct estimation of the post mortem interval (PMI) can be crucial on the success of a forensic investigation. Diverse methods have been used to estimate PMI, considering physical changes that occur after death, such as mortis algor, livor mortis, among others. Degradation after death of dental pulp is a complex process that has not yet been studied thoroughly. It has been described that pulp RNA degradation could be an indicator of PMI, however this study is limited to 6 days. The tooth is the hardest organ of the human body, and within is confined dental pulp. The pulp morphology is defined as a lax conjunctive tissue with great sensory innervation, abundant microcirculation and great presence of groups of cell types. The aim of this study is to describe the potential use of pulp post mortem alterations to estimate PMI, using a new methodology that will allow obtainment of pulp tissue to be used for histomorphological analysis. The current study will identify potential histological indicators in dental pulp tissue to estimate PMI in time intervals of 24h, 1 month, 3 months and 6 months. This study used 26 teeth from individuals with known PMI of 24h, 1 month, 3 months or 6 months. All samples were manipulated with the new methodology (Carrasco, P. and Inostroza C. inventors; Universidad de los Andes, assignee. Forensic identification, post mortem interval estimation and cause of death determination by recovery of dental tissue. United State patent US 61/826,558 23.05.2013) to extract pulp tissue without the destruction of the tooth. The dental pulp tissues obtained were fixed in formalin for the subsequent generation of histological sections, stained with Hematoxylin Eosin and Masson&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s Trichrome. All sections were observed under an optical microscope using magnifications of 10× and 40×. The microscopic analysis of the samples showed a progressive transformation of the cellular components and fibers of dental pulp along PMI. These results allowed creating a chart of qualitative and quantitative parameters to be used on the estimation on PMI based on microscopic degradation of dental pulp. The histological transformations of dental pulp as a function of time can be used as PMI indicators.