Groundwater Quality and Its Impact on Health: A Preliminary Evaluation of Dental Fluorosis in Julimes, Chihuahua, Mexico

Medical Geology can be defined as the branch of geology dealing with the relationship between natural geological aspects and health in organisms (humans and animals), trying to determine the influence of ordinary environmental factors on the geographical distribution of health issues. The main goals of this research were: 1) to evaluate the severity of dental fluorosis (DF) and fluoride concentration in groundwater, used for human consumption, as an indicator of overexposure to fluorides, 2) to estimate the Community Fluorosis Index (CFI) in the Julimes municipality in Chihuahua, Mexico. Nine water samples were collected in four locations in the Julimes area: Hacienda Humboldt, La Regina, Colonia San Jose, and Julimes. Samples were collected and stored in accordance with the Mexican standard (NOM-014-SSA1-1993). All the samples exceeded the maximum permissible limit established in the modified Mexican standard (NOM-127-SSA1-1994) of 1.5 mgL-1 for fluoride in water for human consumption. Average concentrations of 2.74 mgL-1 were found in Colonia San Jose, 2.19 mgL-1 in La Regina, 2.17 mgL-1 in Julimes and 2.06 mgL-1 in Hacienda Humboldt. Dental Fluorosis diagnosis was evaluated using the Dean Index and the CFI applied to residents. A total of 100 inhabitants were examined, with interviews and oral examinations also carried out. Dental Fluorosis prevalence was 92% overall for the municipality, while for Colonia San Jose was of 100%, of 88.89% for Hacienda Humboldt, 88.24% for Julimes and 96% for La Regina. Dental Fluorosis’ severity for the Julimes municipality was distributed as follows: 2% normal, 6% questionable, 8% very mild, 13% mild, 19% moderate, and 52% severe. An average CFI was found for the four localities of 3.02 points, with a very notable impact on public health.

Relationships between daily total fluoride intake and dental fluorosis and dental caries

Objective： To explore the relationships between the daily total intake of fluoride, dental fluorosis and dental caries. Methods： An epidemiological method was used to investigate the daily total intake of fluoride, dental fluorosis, and dental caries among 236 and 290 children aged 8-13 years in a severe endemic area and in a non-fluorosis control area, respectively. The children were divided into eight subgroups according to each child＇ s estimated daily total intake of fluoride. The prevalence of dental fluorosis and dental caries in each group was calculated. Results： As expected, elevated levels of fluoride intake were significantly associated with a higher prevalence of dental fluorosis and an increasing amount of more severe defect dental fluorosis. When the daily total F intake was 2.78 mg/child/day, the prevalence of dental fluorosis was nearly 100%, with the prevalence of defect dental fluorosis increasing with increasing fluoride intake. There was also a significant negative（inverse） dose-response relationship between the daily total intake of fluoride and the overall prevalence of dental caries, the prevalence of which decreased when the daily total intake of fluoride increased up to 3.32 mg/child/day. However, at higher levels of daily total intake of fluoride the prevalence of dental caries increased, giving rise to a U-shaped dose-response relationship curve. Conclusion： It is important to monitor total fluoride exposure and protect children from excessive fluoride intake, especially during the years of tooth development.

Complete oral rehabilitation in a case with severe dental fluorosis

The authors have presented a technique of full occlusal rehabilitation in a case of severe dental fluorosis. In this technique, maxillary and mandibular anterior teeth were simultaneously prepared and restored first. This was followed by simultaneous preparation of maxillary and mandibular posterior teeth that were restored in canine guided occlusion. The technique and sequence followed here is unique and is not available in dental literature. This technique reduces number of appointments while fulfilling all objectives. Periodontal followup over 3 years was satisfactory. A restorative treatment protocol has been devised for fluorosis which will act as a guide for the dental practitioners.

Fluorosis and Dental Caries in the Hydrogeological Environments of Southeastern Communities in the State of Morelos, Mexico

Objectives: 1) To identify the hydrogeological characteristics and fluoride (F-) concentration in drinking water in the southeast of the state of Morelos, Mexico, and 2) to estimate the association between the prevalence and severity of fluorosis, dental caries, and water F-?concentration. Methods: A hydrogeological model was developed to determine the vertical distribution of lithostratigraphic units, rocks’ hydraulic conductivities, and groundwater flow characteristics. A total of 649 schoolchildren from two communities received oral examinations. One community was located in an area with 0.75 mg F-/L in the water supply and the other in an area with 1.6 mg F-/L. Fluorosis was assessed using the Thylstrup & Fejerskov Index (TFI) and the dental caries were assessed with the decayed, missing, and filled teeth (DMFT) index. The region groundwater was extracted from an inter-granular aquifer consisting of volcano clasts of intermediate volcanic chemical composition. Results: Oral examinations showed that 32.35% of the children had severe forms of fluorosis (TFI ≥ 4) in the community with 1.60 mg F-/L and 7.8% in the 0.75 mg F-/L community (p < 0.0001). DMFT = 0.53 (±1.28) and 0.42 (±0.95) in the community with 1.60 mg F-/L and 0.75 mg F-/L, respectively (p = 0.2259). Conclusions: The increasing extraction of underground water was associated with deeper water flows and higher F-?concentration. In the community with high F-concentration, a third of the children showed objectionable levels of fluorosis. Adequate groundwater management actions are required to reduce fluorosis risk.

Fluorine in Water and Dental Fluorosis in a Community of Queretaro State Mexico

The community La Llave, Queretaro State, Mexico, has not been identified by the sanitary authorities as living in a hazard area related to dental fluorosis. However;a high concentration of fluoride is found in their drinking water causing them dental fluorosis. Physical-chemical analysis was carried out in the groundwater sources to determine the prevalence of dental fluorosis risk and caries accordingly to criteria of The World Health Organization, and 154 school children of ages 10 to 13 years were examined. As a result, 1.9 mg/L of fluorine concentration in drinking water was obtained;dental fluorosis presence was detected in the school children with an incidence of about 98%, in 47% of cases severe fluorosis with a dental caries index of 3.06 was traced. The groundwater sources in La Llave community contains fluorine above the limits dictated by Mexican regulations, producing serious repercussions on the health of school children, with an unnaturally high incidence of dental fluorosis.

Prevalence of Dental Fluorosis in Mianwali and Mardan Districts

This study was conducted to assess the level of dental flourosis and Decaying, Missing, Filled Tooth (DMFT) of selected patients, in district Mardan village Rustam and District Mianwali village Harnoli. Study was conducted among people of age 10 - 60 years. The regions of Rustam and Harnoli are known to have high value of fluoride in drinking water. A cross sectional study was conducted among the subjects and the grade of fluorosis was calculated according to Deans Fluorosis Index. DMFT of each subject was also calculated. Total of 100 subjects were selected by simple random sampling technique among the population of Rustam and Harnoli and they were observed for the extent of fluorosis they had along with the number of decayed, missing and filled teeth they had Most of the population uses underground water, the water people commonly used was collected and was tested for amount of fluoride. The results show that there is a strong relationship between fluoride level and extent of fluorosis. Populations of both areas have dental fluorosis due to high concentration of fluoride in drinking water and also comparatively low DMFT because fluoride has caries protective effect.

Self-assembly of differentiated dental pulp stem cells facilitates spheroid human dental organoid formation and prevascularization

BACKGROUND The self-assembly of solid organs from stem cells has the potential to greatly expand the applicability of regenerative medicine.Stem cells can self-organise into microsized organ units,partially modelling tissue function and regeneration.Dental pulp organoids have been used to recapitulate the processes of tooth development and related diseases.However,the lack of vasculature limits the utility of dental pulp organoids.AIM To improve survival and aid in recovery after stem cell transplantation,we demonstrated the three-dimensional(3D)self-assembly of adult stem cell-human dental pulp stem cells(hDPSCs)and endothelial cells(ECs)into a novel type of spheroid-shaped dental pulp organoid in vitro under hypoxia and conditioned medium(CM).METHODS During culture,primary hDPSCs were induced to differentiate into ECs by exposing them to a hypoxic environment and CM.The hypoxic pretreated hDPSCs were then mixed with ECs at specific ratios and conditioned in a 3D environment to produce prevascularized dental pulp organoids.The biological characteristics of the organoids were analysed,and the regulatory pathways associated with angiogenesis were studied.RESULTS The combination of these two agents resulted in prevascularized human dental pulp organoids(Vorganoids)that more closely resembled dental pulp tissue in terms of morphology and function.Single-cell RNA sequencing of dental pulp tissue and RNA sequencing of Vorganoids were integrated to analyse key regulatory pathways associated with angiogenesis.The biomarkers forkhead box protein O1 and fibroblast growth factor 2 were identified to be involved in the regulation of Vorganoids.CONCLUSION In this innovative study,we effectively established an in vitro model of Vorganoids and used it to elucidate new mechanisms of angiogenesis during regeneration,facilitating the development of clinical treatment strategies.

Prospects of 3D Printing Technology in Dental Medicine

With the continuous advancement of technology,the application of 3D printing technology in the field of dental medicine is becoming increasingly widespread.This article aims to explore the current applications and future potential of 3D printing technology in dental medicine and to analyze its benefits and challenges.It first introduces the current state of 3D printing technology in dental implants,crowns,bridges,orthodontics,and maxillofacial surgery.It then discusses the potential applications of 3D printing technology in oral tissue engineering,drug delivery systems,personalized dental prosthetics,and surgical planning.Finally,it analyzes the benefits of 3D printing technology in dental medicine,such as improving treatment accuracy and patient comfort,and shortening treatment times,while also highlighting the challenges faced,such as costs,material choices,and technical limitations.This article aims to provide a reference for professionals in the field of dental medicine and to promote the further application and development of 3D printing technology in this area.

Perception of dental appearance and aesthetic analysis among patients,laypersons and dentists

BACKGROUND Current concepts of beauty are increasingly subjective,influenced by the viewpoints of others.The aim of the study was to evaluate divergences in the perception of dental appearance and smile esthetics among patients,laypersons and dental practitioners.The study goals were to evaluate the influence of age,sex,education and dental specialty on the participants’judgment and to identify the values of different esthetic criteria.Patients sample included 50 patients who responded to a dental appearance questionnaire(DAQ).Two frontal photographs were taken,one during a smile and one with retracted lips.Laypersons and dentists were asked to evaluate both photographs using a Linear Scale from(0-10),where 0 represent(absolutely unaesthetic)and 10 represent(absolutely aesthetic).One-way analysis of variance(ANOVA)and t-test analysis were measured for each group.Most patients in the sample expressed satisfaction with most aspects of their smiles and dental appearance.Among laypersons(including 488 participants),47 pictures“with lips”out of 50 had higher mean aesthetic scores compared to pictures“without lips”.Among the dentist sample,90 dentists’perception towards the esthetic smile and dental appearance for photos“with lips”and“without lips”were the same for 23 out of 50 patients.Perception of smile aesthetics differed between patients,laypersons and dentists.Several factors can contribute to shape the perception of smile aesthetic.AIM To compare the perception of dental aesthetic among patients,laypersons,and professional dentists,to evaluate the impact of age,sex,educational background,and income on the judgments made by laypersons,to assess the variations in experience,specialty,age,and sex on professional dentists’judgment,and to evaluate the role of lips,skin shade and tooth shade in different participants’judgments.METHODS Patients sample included 50 patients who responded to DAQ.Two frontal photographs were taken:one during a smile and one with retracted lips.Laypersons and dentists were asked to evaluate both photographs using a Linear Scale from(0-10),where 0 represent(absolutely unaesthetic)and 10 represent(absolutely aesthetic).One-way ANOVA and t-test analysis were measured for each group.RESULTS Most patients in the sample expressed satisfaction with most aspects of their smiles and dental appearance.Among laypersons(including 488 participants),47 pictures“with lips”out of 50 had higher mean aesthetic scores compared to pictures“without lips”.Whereas among the dentist sample,90 dentists’perception towards the esthetic smile and dental appearance for photos“with lips”and“without lips”were the same for 23 out of 50 patients.Perception of smile aesthetics differed between patients,laypersons and dentists.CONCLUSION Several factors can contribute to shape the perception of smile aesthetic.

Conditioned medium from human dental pulp stem cells treats spinal cord injury by inhibiting microglial pyroptosis

Human dental pulp stem cell transplantation has been shown to be an effective therapeutic strategy for spinal cord injury.However,whether the human dental pulp stem cell secretome can contribute to functional recovery after spinal cord injury remains unclear.In the present study,we established a rat model of spinal cord injury based on impact injury from a dropped weight and then intraperitoneally injected the rats with conditioned medium from human dental pulp stem cells.We found that the conditioned medium effectively promoted the recovery of sensory and motor functions in rats with spinal cord injury,decreased expression of the microglial pyroptosis markers NLRP3,GSDMD,caspase-1,and interleukin-1β,promoted axonal and myelin regeneration,and inhibited the formation of glial scars.In addition,in a lipopolysaccharide-induced BV2 microglia model,conditioned medium from human dental pulp stem cells protected cells from pyroptosis by inhibiting the NLRP3/caspase-1/interleukin-1βpathway.These results indicate that conditioned medium from human dental pulp stem cells can reduce microglial pyroptosis by inhibiting the NLRP3/caspase-1/interleukin-1βpathway,thereby promoting the recovery of neurological function after spinal cord injury.Therefore,conditioned medium from human dental pulp stem cells may become an alternative therapy for spinal cord injury.

Classification and detection of dental images using meta-learning

Meta-learning of dental X-rays is a machine learning technique that can be used to train models to perform new tasks quickly and with minimal input.Instead of just memorizing a task,this is accomplished through teaching a model how to learn.Algorithms for meta-learning are typically trained on a collection of training problems,each of which has a limited number of labelled instances.Multiple Xray classification tasks,including the detection of pneumonia,coronavirus disease 2019,and other disorders,have demonstrated the effectiveness of meta-learning.Meta-learning has the benefit of allowing models to be trained on dental X-ray datasets that are too few for more conventional machine learning methods.Due to the high cost and lengthy collection process associated with dental imaging datasets,this is significant for dental X-ray classification jobs.The ability to train models that are more resistant to fresh input is another benefit of meta-learning.

Adjuvant postbiotic administration improves dental caries prognosis by restoring the oral microbiota

Conventional filling therapy fails to fundamentally reduce oral cariogenic bacteria.Thus,oral microbiota follow-up intervention after filling would be necessary for improving dental caries prognosis.We recruited 9 caries-free individuals,and 89 dental caries subjects(5 dropouts).Eighty-nine patients were randomized into three groups:caries(n=8;no treatment),control(n=40;filling),and postbiotics(n=41;filling and 14-day Probio-Eco®intervention).Salivary samples were collected at 0 day(after filling)and 14 days.Our results showed that the diversity of dental caries oral microbiota was significantly increased compared with healthy subjects,and filling could restore a healthier oral microbiota partially and temporarily.Thepostbiotics intervention keeps a low alpha-diversity.Co-occurrence network analysis showed that a more stable oral microbiota structure after postbiotics intervention.Taxonomic and functional annotation of the microbiota revealed that postbiotics co-treatment significantly:increased the relative abundance of Pseudomonas and P.reactans,decreased the relative abundance of Prevotella shahii,and enriched the energy metabolism-related pathways.BugBase-predicted phenotypes inferred to an oral microbiota with decreased potential pathogenic bacteria and increased oxidative stress-tolerant bacteria after postbiotics intervention.Collectively,it suggested that postbiotics co-treatment could be a promising strategy that restores the oral microecological balance for dental caries.

The Dental Absces: Epidemiological, Clinical and Therapeutic Aspects at the Dental Office of the Prefectoral Hospital of Siguiri in 2021 in the Republic of Guinea

Introduction: A dental abscess is a collection of pus located in the tissues surrounding the tooth roots. The aim of this work is to describe the epidemiological, clinical and therapeutic aspects of dental abscess in order to contribute to the management of patients suffering from this pathology at the dental office of the Siguiri Prefectural Hospital. Methods: This was a retrospective descriptive study lasting 2 months carried out on the registers of patients received between October 2017 and October 2020 at the dental office of the Siguiri prefectural hospital. Results: 4690 patient files were identified during the study, of which 546 met our selection criteria, i.e. a frequency of 11.64%. During our study, we found 51.83% female compared to 48.17% male, i.e. a sex ratio of 1.07. 54.21% of our patients resided in rural communes compared to 45.76% for the urban commune. The age group most affected was that of 16 - 25 years old or 33.69%. Swelling, pain and functional discomfort were the reasons for consultation in all patients followed by other associated reasons in 41.21%. The premolar-molar group was the most represented, i.e. 86.70%. 80.04% of our patients presented an acute abscess compared to 19.96% of chronic cases. Antibiotic, analgesic, and mouthwash were prescribed in all patients during the study followed by 96.37% of cases prescription of steroidal anti-inflammatory drugs. 93.04% of causal teeth were extracted followed by 6.78% of cases of conservative treatment in our study. In our study, 80.95% of our patients had a good post-therapeutic outcome. Conclusion: This study highlights the need for prevention and early management of dental abscesses.

Study on the Treatment of Reusable Care Equipment in Public and Private Dental Practices in the City of Conakry

Introduction: The effective sterilization of reusable instruments in dental care is a crucial issue for public health. The aim of this study was to contribute to improving the processing conditions of reusable care instruments in dental practices in the city of Conakry. Methods: We carried out an observational and cross-sectional study of a descriptive type lasting three months from January to April 2022 in public and private dental practices approved for activities. Results: A total of 48 dental practices were surveyed, including 7 public and 41 private. 91.67% had the trays for instrument sterilization. 77.08% had a sterilization device. The instruments were: cleaned 100%, disinfected 70.83%, sterilized 20.83% and 20.83% reused the instruments without being sterilized. 72.97% sterilized the instruments at the end of the day. 50% of respondents declared that there were six stages of sterilization. According to standard standards, one dental office or 2.70% respected the normal sterilization process and 13.51% respected the duration and temperature. The storage quality was inadequate at 95.83%. 95.83% of dental practices are at high risk of contamination. The difficulties encountered by staff in daily activity during our study were lack of hygienists (87.50%), insufficient material resources (58.33%), and lack of protocol procedure (45.83%). Conclusion: This study allowed us to note shortcomings of dental surgeons in the process of processing reusable care materials.

Prevalence of Dental Anomalies among Patients Who Report to the Komfo Anokye Teaching Hospital’s Orthodontic Clinic

BACKGROUND: Dental anomalies are variations from the established well-known general anatomy and morphology of the tooth as a result of disturbances during tooth formation. They can be developmental, congenital, or acquired and may be localized to a single tooth or involve systemic conditions. AIM: To evaluate the prevalence of dental anomalies in patients who report to the Komfo Anokye Teaching Hospital (KATH) dental clinics. METHOD: A descriptive cross-sectional design was used with a sample size of 92 patients aged 18 or older, obtained through convenience sampling. Data analysis was performed using SPSS version 26.0. RESULTS: The study included 92 patients aged 18 to 72 years, with 47.8% males and 52.2% females. Dental anomalies were observed in 51.1% of participants, with a higher prevalence in females (55.3%). The most common anomalies were diastema (48.3%), impacted teeth (22.0%), dilaceration (11.9%), and peg-shaped lateral teeth (6.8%). CONCLUSION: This study highlights the importance of conducting thorough dental examinations to identify and address dental anomalies, which may have implications for treatment. Early detection and correction of these anomalies are crucial to prevent future complications.

Prescription of Antibiotics in Oral-Dental Practice: Case of the Dental Practice at the Coyah Prefectoral Hospital in the Republic of Guinea 2019-2020

Introduction: Antibiotics are medications that have the property of destroying bacteria or preventing their proliferation. The aim of this study is to evaluate antibiotic prescriptions in oral practice: case of the dental office of the Coyah prefectural hospital. Methods: This is a descriptive cross-sectional study which took place from August 2019 to January 2020 inclusive, i.e. a duration of six (6) months, during which 400 patients were identified. Results: Our results revealed a frequency of 70% of antibiotic prescriptions. The 16 - 25-year-old age group was the most dismayed, at 47.25% and with an average age of 27.78 years. Men represent 55% compared to 45% of women, i.e. sex ratio of 1.22. Pulp gangrene was the most common pathology in 55% of cases. The most prescribed family was Betalactamine or 61.71% and the most represented molecules were Amoxicillin with 56.10% followed by Metronidazole or 34.16%. And the favorable prognosis was observed in 99% of cases. Conclusion: Prescription should always be based on the benefit-risk ratio. This prescription by practitioners aims to prevent or eradicate infections and pain in order to obtain a good therapeutic result.

Associated Factors of Dental Caries among Intellectually Disabled Persons in Specialized Centers of Ouagadougou

Introduction: Several studies have reported a high prevalence of dental caries among the people with intellectual disabilities. In order to identify the sociodemographic, clinical and behavioral factors associated with it, we conducted a study in specialized centers welcoming subjects with intellectual disabilities in Ouagadougou (Burkina Faso). Objectives: The objective of this study was to identify the factors associated with it among intellectually disabled in the specialized centers of Ouagadougou. Methods: The study was cross-sectional analytical. The survey took place between November 2020 and January 2021 among people with intellectual disabilities in reception centers receiving a grant from the Ministry of Women, National Solidarity, Family and Humanitarian Action. Data collection was done with a questionnaire adapted from the World Health Organization (WHO) and dental caries was recorded according to WHO criteria in decayed, missing or filled teeth generating a DMF index. Logistic regression was used to identify factors associated with the presence of dental caries. Results: A total of 193 participants were included in the study with an average age of 12.4 years ± 7.9. The overall prevalence of caries measured with the DMF index was 58.6% 95% CI [51.2 - 65.8]. It was significantly associated with female gender (OR = 4.2;95% CI [1.4 - 12.5], p = 0,01), the clinical form of mental illness (epilepsy OR = 3.8 95%;CI [1.2 - 12.7], p = 0,02, trisomy OR = 5.0;95% CI [1.1 - 22.9], p = 0,03, motor autonomy OR = 0.2 95%;CI [0.1 - 0.7], p = 0,01) and at use of toothpaste OR = 9.33;95% CI [1.05 - 82.7], p = 0,04. Conclusion: Dental caries remains a very present pathology in most people living with an intellectual disability. Also, it is necessary to put in place prevention strategies to reduce its prevalence and improve access to oral care for these people.

An overview of autophagy in the differentiation of dental stem cells

Dental stem cells(DSCs)have attracted significant interest as autologous stem cells since they are easily accessible and give a minimal immune response.These properties and their ability to both maintain self-renewal and undergo multi-lineage differentiation establish them as key players in regenerative medicine.While many regulatory factors determine the differentiation trajectory of DSCs,prior research has predominantly been based on genetic,epigenetic,and molecular aspects.Recent evidence suggests that DSC differentiation can also be influenced by autophagy,a highly conserved cellular process responsible for maintaining cellular and tissue homeostasis under various stress conditions.This comprehensive review endeavors to elucidate the intricate regulatory mechanism and relationship between autophagy and DSC differentiation.To achieve this goal,we dissect the intricacies of autophagy and its mechanisms.Subsequently,we elucidate its pivotal roles in impacting DSC differentiation,including osteo/odontogenic,neurogenic,and angiogenic trajectories.Furthermore,we reveal the regulatory factors that govern autophagy in DSC lineage commitment,including scaffold materials,pharmaceutical cues,and the extrinsic milieu.The implications of this review are far-reaching,underpinning the potential to wield autophagy as a regulatory tool to expedite DSC-directed differentiation and thereby promote the application of DSCs within the realm of regenerative medicine.

RPLP0/TBP are the most stable reference genes for human dental pulp stem cells under osteogenic differentiation

BACKGROUND Validation of the reference gene(RG)stability during experimental analyses is essential for correct quantitative real-time polymerase chain reaction(RT-qPCR)data normalisation.Commonly,in an unreliable way,several studies use genes involved in essential cellular functions[glyceraldehyde-3-phosphate dehydro-genase(GAPDH),18S rRNA,andβ-actin]without paying attention to whether they are suitable for such experimental conditions or the reason for choosing such genes.Furthermore,such studies use only one gene when Minimum Information for Publication of Quantitative Real-Time PCR Experiments guidelines recom-mend two or more genes.It impacts the credibility of these studies and causes dis-tortions in the gene expression findings.For tissue engineering,the accuracy of gene expression drives the best experimental or therapeutical approaches.We cultivated DPSCs under two conditions:Undifferentiated and osteogenic dif-ferentiation,both for 35 d.We evaluated the gene expression of 10 candidates for RGs[ribosomal protein,large,P0(RPLP0),TATA-binding protein(TBP),GAPDH,actin beta(ACTB),tubulin(TUB),aminolevulinic acid synthase 1(ALAS1),tyro-sine 3-monooxygenase/tryptophan 5-monooxygenase activation protein,zeta(YWHAZ),eukaryotic translational elongation factor 1 alpha(EF1a),succinate dehydrogenase complex,subunit A,flavoprotein(SDHA),and beta-2-micro-globulin(B2M)]every 7 d(1,7,14,21,28,and 35 d)by RT-qPCR.The data were analysed by the four main algorithms,ΔCt method,geNorm,NormFinder,and BestKeeper and ranked by the RefFinder method.We subdivided the samples into eight subgroups.RESULTS All of the data sets from clonogenic and osteogenic samples were analysed using the RefFinder algorithm.The final ranking showed RPLP0/TBP as the two most stable RGs and TUB/B2M as the two least stable RGs.Either theΔCt method or NormFinder analysis showed TBP/RPLP0 as the two most stable genes.However,geNorm analysis showed RPLP0/EF1αin the first place.These algorithms’two least stable RGs were B2M/GAPDH.For BestKeeper,ALAS1 was ranked as the most stable RG,and SDHA as the least stable RG.The pair RPLP0/TBP was detected in most subgroups as the most stable RGs,following the RefFinfer ranking.CONCLUSION For the first time,we show that RPLP0/TBP are the most stable RGs,whereas TUB/B2M are unstable RGs for long-term osteogenic differentiation of human DPSCs in traditional monolayers.