RESUMEN
OBJECTIVES: To identify and report the current landmarks used for measuring gingival thickness (GT) in healthy maxillary anterior teeth. MATERIAL AND METHODS: The protocol of this Preferred Reporting Items of Systematic Reviews and Meta-Analyses (PRISMA) 2020-compliant systematic review was registered in PROSPERO. A literature search was conducted to identify articles that met the eligibility criteria published up to 2022. The methods of assessing gingival thickness and the landmarks adopted on the studies were described. Primary outcomes were identified, and the frequency of reporting in the selected articles was calculated. Additionally, risk-of-bias assessments were performed for individual articles. RESULTS: Fifty-eight articles (34 with low risk of bias and 24 with medium risk of bias) were selected. A total of 3638 individuals had their gingival thickness measured. Thirty-nine different landmarks were adopted in the studies. Fifty-six articles with 22 landmarks were included in the meta-analysis. A higher heterogeneity was found between the studies (GT ranged from 0.48 to 2.59 mm, mean GT 1.074; 95% CI: 1.024-1.104). The 3 most used landmarks were 2 mm from gingival margin (10 studies, mean GT 1.170 mm, 95% CI: 1.085-1.254), bone crest (9 studies, mean GT 1.01 mm; 95% CI: 0.937-1.083), and cemento-enamel junction (7 studies, mean GT 1.172 mm; 95% CI: 1.105, 1.239). CONCLUSIONS: Within the limits of this study, a large heterogeneity in GT was found, and there was no consensus on the ideal landmark for GT measurement. CLINICAL RELEVANCE: The landmark 2 mm from gingival margin, located at attached gingiva, can be used for GT measurement by clinical and image-based devices. This is an important step for a quantitative instead of a qualitative evaluation of phenotypes.
Asunto(s)
Encía , Maxilar , Diente , Tomografía Computarizada de Haz Cónico/métodos , Encía/citología , Maxilar/citología , Cuello del DienteRESUMEN
Neural crest cells (NCCs) within the mandibular and maxillary prominences of the first pharyngeal arch are initially competent to respond to signals from either region. However, mechanisms that are only partially understood establish developmental tissue boundaries to ensure spatially correct patterning. In the 'hinge and caps' model of facial development, signals from both ventral prominences (the caps) pattern the adjacent tissues whereas the intervening region, referred to as the maxillomandibular junction (the hinge), maintains separation of the mandibular and maxillary domains. One cap signal is GATA3, a member of the GATA family of zinc-finger transcription factors with a distinct expression pattern in the ventral-most part of the mandibular and maxillary portions of the first arch. Here, we show that disruption of Gata3 in mouse embryos leads to craniofacial microsomia and syngnathia (bony fusion of the upper and lower jaws) that results from changes in BMP4 and FGF8 gene regulatory networks within NCCs near the maxillomandibular junction. GATA3 is thus a crucial component in establishing the network of factors that functionally separate the upper and lower jaws during development.
Asunto(s)
Tipificación del Cuerpo , Cara/embriología , Factor de Transcripción GATA3/metabolismo , Animales , Región Branquial/citología , Región Branquial/embriología , Región Branquial/metabolismo , Muerte Celular , Proliferación Celular , Anomalías Craneofaciales/embriología , Anomalías Craneofaciales/genética , Anomalías Craneofaciales/metabolismo , Embrión de Mamíferos , Factor de Transcripción GATA3/genética , Regulación del Desarrollo de la Expresión Génica , Mandíbula/citología , Mandíbula/embriología , Maxilar/citología , Maxilar/embriología , Ratones , Morfogénesis , Cresta Neural/citología , Cresta Neural/embriología , Cresta Neural/metabolismoRESUMEN
BACKGROUND: Multipotent mesenchymal stromal cells (MSCs) can be isolated from numerous tissues and are attractive candidates for therapeutic clinical applications due to their immunomodulatory and pro-regenerative capacity. Although the minimum criteria for defining MSCs have been defined, their characteristics are known to vary depending on their tissue of origin. RESULTS: We isolated and characterized human MSCs from three different bones (ilium (I-MSCs), maxilla (Mx-MSCs) and mandible (Md-MSCs)) and proceeded with next generation RNA-sequencing. Furthermore, to investigate the gene expression profiles among other cell types, we obtained RNA-seq data of human embryonic stem cells (ESCs) and several types of MSCs (periodontal ligament-derived MSCs, bone marrow-derived MSCs, and ESCs-derived MSCs) from the Sequence Reads Archive and analyzed the transcriptome profile. We found that MSCs derived from tissues of the maxillofacial region, such as the jaw bone and periodontal ligament, were HOX-negative, while those derived from other tissues were HOX-positive. We also identified that MSX1, LHX8, and BARX1, an essential regulator of craniofacial development, were strongly expressed in maxillofacial tissue-derived MSCs. Although MSCs may be divided into two distinct groups, the cells originated from over the neck or not, on the basis of differences in gene expression profile, the expression patterns of all CD antigen genes were similar among different type of MSCs, except for ESCs. CONCLUSIONS: Our findings suggest that MSCs from different anatomical locations, despite meeting general characterization criteria, have remarkable differences in gene expression and positional memory. Although stromal cells from different anatomical sources are generally categorized as MSCs, their differentiation potential and biological functions vary. We suggested that MSCs may retain an original tissue memory about the developmental process, including gene expression profiles. This could have an important impact when choosing an appropriate cell source for regenerative therapy using MSCs.
Asunto(s)
Perfilación de la Expresión Génica/métodos , Redes Reguladoras de Genes , Ilion/citología , Mandíbula/citología , Maxilar/citología , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Regulación de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Proteínas de Homeodominio/genética , Humanos , Ilion/química , Mandíbula/química , Maxilar/química , Células Madre Mesenquimatosas/química , Células Madre Mesenquimatosas/citología , Especificidad de Órganos , Análisis de Secuencia de ARN/métodos , Secuenciación del ExomaRESUMEN
During mammalian palatogenesis, cranial neural crest-derived mesenchymal cells undergo osteogenic differentiation and form the hard palate, which is divided into palatine process of the maxilla and the palatine. However, it remains unknown whether these bony structures originate from the same cell lineage and how the hard palate is patterned at the molecular level. Using mice, here we report that deficiency in Shox2 (short stature homeobox 2), a transcriptional regulator whose expression is restricted to the anterior palatal mesenchyme, leads to a defective palatine process of the maxilla but does not affect the palatine. Shox2 overexpression in palatal mesenchyme resulted in a hyperplastic palatine process of the maxilla and a hypoplastic palatine. RNA sequencing and assay for transposase-accessible chromatin-sequencing analyses revealed that Shox2 controls the expression of pattern specification and skeletogenic genes associated with accessible chromatin in the anterior palate. This highlighted a lineage-autonomous function of Shox2 in patterning and osteogenesis of the hard palate. H3K27ac ChIP-Seq and transient transgenic enhancer assays revealed that Shox2 binds distal-acting cis-regulatory elements in an anterior palate-specific manner. Our results suggest that the palatine process of the maxilla and palatine arise from different cell lineages and differ in ossification mechanisms. Shox2 evidently controls osteogenesis of a cell lineage and contributes to the palatine process of the maxilla by interacting with distal cis-regulatory elements to regulate skeletogenic gene expression and to pattern the hard palate. Genome-wide Shox2 occupancy in the developing palate may provide a marker for identifying active anterior palate-specific gene enhancers.
Asunto(s)
Diferenciación Celular/genética , Proteínas de Homeodominio/genética , Osteogénesis/genética , Paladar Duro/metabolismo , Animales , Tipificación del Cuerpo/genética , Linaje de la Célula/genética , Femenino , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/metabolismo , Humanos , Maxilar/citología , Maxilar/embriología , Maxilar/metabolismo , Ratones Noqueados , Ratones Transgénicos , Paladar Duro/citología , Paladar Duro/embriología , Transducción de Señal/genéticaRESUMEN
MicroRNAs (miRNAs) posttranscriptionally regulate gene expression by targeting the 3' untranslated region (UTR) of target genes, and serve diverse roles in cell proliferation, differentiation and apoptosis. However, the association between miR1623p and 3phosphoinositidedependent protein kinase1 (PDPK1) in nonsyndromic cleft lip (NSCL) remains unclear. In the present study, a luciferase activity assay indicated that miR1623p negatively regulated PDPK1 in maxillary primordium mesenchymal cells (MPMCs). In addition, it was confirmed that the expression levels of miR1623p was markedly increased in cleft lip tissues compared with those in adjacent normal lip tissues. A negative correlation between miR1623p and PDPK1 in cleft lip tissues was observed. Furthermore, miR1623p inhibited cell proliferation and migration, and induced apoptosis of MPMCs via repressing PDPK1. Finally, miR1623p exerted its suppressive role in MPMCs by inhibiting the PDPK1/protein kinase B signaling pathway. These results indicate that miR1623p may inhibit cell proliferation and migration, and promote apoptosis in MPMCs through repression of PDPK1 and may be a potential target for future clinical prevention and treatment of NSCL.
Asunto(s)
Proteínas Quinasas Dependientes de 3-Fosfoinosítido/genética , Apoptosis/genética , Maxilar/citología , MicroARNs/genética , Interferencia de ARN , Regiones no Traducidas 3' , Proteínas Quinasas Dependientes de 3-Fosfoinosítido/metabolismo , Ciclo Celular , Puntos de Control del Ciclo Celular/genética , Diferenciación Celular , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Regulación de la Expresión Génica , Técnicas de Silenciamiento del Gen , Humanos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de SeñalRESUMEN
Gnathostome jaws derive from the first pharyngeal arch (PA1), a complex structure constituted by Neural Crest Cells (NCCs), mesodermal, ectodermal and endodermal cells. Here, to determine the regionalized morphogenetic impact of Dlx5/6 expression, we specifically target their inactivation or overexpression to NCCs. NCC-specific Dlx5/6 inactivation (NCC∆Dlx5/6) generates severely hypomorphic lower jaws that present typical maxillary traits. Therefore, differently from Dlx5/6 null-embryos, the upper and the lower jaws of NCC∆Dlx5/6 mice present a different size. Reciprocally, forced Dlx5 expression in maxillary NCCs provokes the appearance of distinct mandibular characters in the upper jaw. We conclude that: (1) Dlx5/6 activation in NCCs invariably determines lower jaw identity; (2) the morphogenetic processes that generate functional matching jaws depend on the harmonization of Dlx5/6 expression in NCCs and in distinct ectodermal territories. The co-evolution of synergistic opposing jaws requires the coordination of distinct regulatory pathways involving the same transcription factors in distant embryonic territories.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/biosíntesis , Mandíbula/embriología , Maxilar/embriología , Cresta Neural/embriología , Animales , Proteínas de Homeodominio/genética , Mandíbula/citología , Maxilar/citología , Ratones , Ratones Mutantes , Cresta Neural/citologíaRESUMEN
PURPOSE: To compare the differences between the osteogenic development in vivo and osteogenic differentiation in vitro of mouse maxillary primordium mesenchymal cells (MPMCs). METHODS: E10.5 and E17.5 mouse MPMCs were cultured in vitro to observe cell morphology. E10.5 primary MPMCs, after culturing in vitro for 3 days, were cultured in osteogenic differentiation in vitro for another 7 days. Then immunofluorescence and qPCR were used to compare the difference of osteogenic differentiation with E17.5 primary MPMCs cultured in vitro for 3 days. SPSS 20.0 software package was used for independent samples t test. RESULTS: E10.5 and E17.5 mouse MPMCs adhered to dish when cultured in vitro, and the cells exhibited polygonal or oval shape. The proliferation of E17.5 mouse MPMCs was faster than that of E10.5 MPMCs. After 7 days of osteogenic induction, the expression of Runx2 and OCN proteins, two osteogenic markers, in E10.5 mouse MPMCs was similar to E17.5 cells without osteogenic induction. The mRNA expression of Runx2, OCN and OPN also showed similar expression patterns, and there was no significant difference in the calcium nodules formation between E10.5 MPMCs and E17.5 MPMCs. CONCLUSIONS: In vitro osteogenic induction of E10.5 mouse MPMCs can mimic osteogenic development process of MPMCs in vivo, and provide a suitable cell model for the study of jaw development.
Asunto(s)
Diferenciación Celular , Maxilar , Células Madre Mesenquimatosas , Osteogénesis , Animales , Células Cultivadas , Maxilar/citología , RatonesRESUMEN
La osteonecrosis de maxilar asociada a aminobisfosfonatos (BRONJ) constituye un efecto secundario del tratamiento crónico con los más potentes. Un modelo experimental permitiría determinar la patogenia de dicha alteración. La oveja presenta características orales y del metabolismo óseo similar al humano y permite realizar manipulaciones bucales. Se evaluaron cambios clínicos, remodelación ósea y masa ósea maxilar en ovejas hembras adultas tratadas con zolendronato (ZOL), durante 22 meses y utilizando dosis equivalente al tratamiento de neoplasias. Seis ovariectomizadas (OVX) recibieron ZOL; 5 OVX y 4 SHAM (control) recibieron solución fisiológica. Al inicio, 4 y 22 meses se evaluó calcemia, fosfatemia, crosslaps (CTX) y fosfatasa alcalina ósea. Al final, se evaluó contenido mineral óseo de la hemimandíbula superior (CMO: mg/cm2). Al final del estudio, CTX disminuyó significativamente en ZOL (p<0,05) sin diferencias entre SHAM y OVX. En maxilar, los contenidos de Ca y P (g/g tejido) y CMO (g/cm2 ) disminuyeron en OVX vs. SHAM (p<0,05) y solo Ca y CMO respecto de ZOL (p<0,05). ZOL incrementó el contenido de Ca y CMO, mientras que el de P permaneció significativamente disminuido respecto de SHAM. La sobrevida en SHAM y OVX fue del 100% y en ZOL 77% (2 muertes); 2 ovejas del grupo ZOL presentaron necrosis de maxilar. Conclusiones: fue posible obtener desarrollo de BRONJ por tratamiento crónico con ZOL, el cual redujo notablemente la resorción y, según la relación Ca/P, posiblemente haya afectado la mineralización ósea. (AU)
Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a complication of chronic treatment with the most powerful aminobisphosphonates (BPs). An experimental animal model would allow to determine the pathogenesis of this complication. Ewes exhibit similar oral cavity characteristics and bone metabolism as humans, and they are suitable for oral cavity interventions. We examined herein the clinical manifestations, bone remodeling status, and maxillary bone mass in adult female ewes treated with zoledronate (ZOL) for 22 months. Six ovariectomized (OVX) ewes received ZOL; and 5 OVX and 4 SHAM animals received saline solution. At the start of the experiment, and at the 4 and 22 month-time points serum Ca, P, crosslaps (CTX), and bone alkaline phosphatase were measured. Bone mineral content (BMC) of the superior hemimandible was measured at the end of the experiment. At this time point, CTX was significantly decreased only in the ZOL group (p<0.05). Ca and P content (g/g tissue) and BMC in the mandible were significantly decreased in the OVX group compared to SHAM animals (p<0.05) and only Ca content and BMC were decreased when compared to ZOL (p<0.05). ZOL treatment increased the Ca content and BMC, whereas the P content remained low compared to the SHAM group (p<0.05). All ewes from the SHAM and OVX groups and 77% of the animals from the ZOL group survived until the end of the experiment, whereas two ewes of ZOL group exhibited BRONJ. Conclusion: under our experimental conditions, it was possible to induce BRONJ by the chronic ZOL administration, which in turn induced a high reduction in bone resorption as well as possibly impaired bone mineralization, based on the Ca/P ratio in the mandible. (AU)
Asunto(s)
Animales , Difosfonatos/efectos adversos , Osteonecrosis de los Maxilares Asociada a Difosfonatos/patología , Ácido Zoledrónico/efectos adversos , Extracción Dental , Enfermedades Óseas Metabólicas/inducido químicamente , Ovinos/metabolismo , Ovinos/sangre , Biomarcadores/sangre , Densidad Ósea/efectos de los fármacos , Remodelación Ósea/efectos de los fármacos , Densitometría , Desarrollo Experimental , Osteonecrosis de los Maxilares Asociada a Difosfonatos/etiología , Osteonecrosis de los Maxilares Asociada a Difosfonatos/inmunología , Osteonecrosis de los Maxilares Asociada a Difosfonatos/prevención & control , Ácido Zoledrónico/administración & dosificación , Glucocorticoides/uso terapéutico , Analgésicos/uso terapéutico , Ilion/citología , Anestésicos Disociativos/uso terapéutico , Lidocaína/uso terapéutico , Maxilar/citología , Maxilar/efectos de los fármacos , Maxilar/metabolismo , Maxilar/diagnóstico por imagen , Antibacterianos/uso terapéuticoRESUMEN
BACKGROUND: Dentigerous cyst (DC) occurs in approximately 20% of jaw cysts, being the second major common odontogenic cyst, after radicular cyst. This oral lesion has the ability to destroy maxillary bones and could be the origin of several odontogenic tumors. However, molecules implicated in its pathogenesis as well as those involved in its neoplastic transformation remain unknown. Here, we established a cell population derived from a DC as an in vitro model for the study of this oral lesion. METHODS: Cell culture was performed from a DC from a 44-year-old male. Cells were cultured at 37°C in DMEM/F12 medium containing 10% fetal bovine serum. Expression of epithelial markers was analyzed by Western blot and immunofluorescence. Ultrastructural characterization was carried out by transmission electron microscopy. Conditioned media were obtained and characterized by zymography and Western blot. RESULTS: Cells showed spindle-shaped morphology, but they express epithelial markers, such as cytokeratins and the odontogenic ameloblast-associated protein. The ultrastructural analysis showed well-formed desmosomes present in adhering contiguous cells, confirming the epithelial lineage of this cell population. Cells also contain several vesicles adjacent to plasma membrane, suggesting an active secretion. Indeed, the analysis of the conditioned medium revealed the presence of several secreted proteins, among them the matrix metalloproteinase-2. CONCLUSIONS: Our work provides a useful model to identify the molecular mechanisms involved in the pathogenesis of DC.
Asunto(s)
Quiste Dentígero/patología , Enfermedades Maxilares/patología , Adulto , Western Blotting , Células Cultivadas , Técnica del Anticuerpo Fluorescente , Humanos , Masculino , Maxilar/citología , Maxilar/patologíaRESUMEN
Previous reports have mainly focused on the behavioral responses of human periodontal ligament stem cells (hPDLSCs) in interaction with tibia bone marrow stromal cells (BMSCs). However, there is little study on the biologic features of hPDLSCs under the induction of maxilla BMSCs (M-BMSCs) at different phases of osteogenic differentiation. We hypothesized that M-BMSCs undergoing osteogenic differentiation acted on the proliferation, differentiation, and bone-forming capacity of hPDLSCs. In this paper, primary hPDLSCs and human M-BMSCs (hM-BMSCs) were expanded in vitro. After screening of surface markers for characterization, hPDLSCs were cocultured with different phases of differentiating hM-BMSCs. Cell proliferation and alkaline phosphatase activity were examined, and mineralization-associated markers such as osteocalcin and runt-related transcription factor 2 of hPDLSCs in coculture with uninduced/osteoinduced hM-BMSCs were evaluated. hPDLSCs in hM-BMSCs-conditioned medium (hM-BMSCs-CM) group showed a reduction in proliferation compared with untreated hPDLSCs, while osteoinduced hM-BMSCs for 10 day-conditioned medium (hM-BMSCs-CM-10ds) and osteoinduced hM-BMSCs for 15 day-conditioned medium (hM-BMSCs-CM-15ds) enhance the proliferation of hPDLSCs. hM-BMSCs of separate differentiation stages temporarily inhibited osteogenesis of hPDLSCs in the early days. Upon extending time periods, uninduced/osteoinduced hM-BMSCs markedly enhanced osteogenesis of hPDLSCs to different degrees. The transplantation results showed hM-BMSCs-CM-15ds treatment promoted tissue regeneration to generate cementum/periodontal ligament-like structure characterized by hard-tissue formation. This research supported the notion that hM-BMSCs triggered osteogenesis of hPDLSCs suggesting important implications for periodontal engineering.
Asunto(s)
Diferenciación Celular , Proliferación Celular/efectos de los fármacos , Técnicas de Cocultivo , Medios de Cultivo Condicionados , Maxilar/citología , Células Madre Mesenquimatosas/citología , Osteogénesis , Ligamento Periodontal/citología , Células Madre/citología , Células Madre/metabolismo , Adolescente , Células Cultivadas , Niño , Medios de Cultivo Condicionados/farmacología , Humanos , Osteogénesis/efectos de los fármacosRESUMEN
Tissue engineering strategies hold great potential for alveolar cleft reconstruction. Bone marrow stromal cells (BMSCs) from iliac crest and craniofacial regions are candidate seeding cells with site-specific characteristics and bone-repairing properties. Craniofacial BMSCs seem to possess stronger multipotency and osteogenic capabilities than BMSCs isolated from iliac crest. However, the angiogenic capabilities of these two type cell is rarely reported. We obtained human BMSCs (hBMSCs) of maxilla (M-hBMSCs) and iliac crest (I-hBMSCs) from same alveolar cleft patients to investigate the agiogenic variations using co-culture system with human umbilical vein endothelial cells (HUVECs). From in vitro comparison, M-hBMSCs allowed HUVECs to form more tube-like structures and sprouting angiogenesis by tube formation assays and 3D fibrin vasculogenic assay, respectively. By transplantation in vivo, M-hBMSCs enhanced larger size vessel like structures distributed the entire implants compared with I-hBMSCs. Western blotting was used to assess the angiogenesis related factors including hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). The results showed a significant higher expression of bFGF protein in M-hBMSCs and HUVECs co-culture system both in vitro and in vivo. As bFGF could promote migration and proliferation of endothelial cells, scratch wound healing and transwell migration assays were performed as well as MTT assays and cell cycle analysis. The data suggested the effect of M-hBMSCs on HUVECs was stronger than I-hBMSCs. Taken together, these results indicated that craniofacial BMSCs seemed to have greater angiogenesis capability than iliac crest BMSCs and this might be associated with the different levels of bFGF protein expression in co-culture system.
Asunto(s)
Huesos/citología , Labio Leporino/patología , Células Madre Mesenquimatosas/citología , Neovascularización Fisiológica , Adipogénesis , Adolescente , Diferenciación Celular , Movimiento Celular , Proliferación Celular , Niño , Técnicas de Cocultivo , Femenino , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Células Endoteliales de la Vena Umbilical Humana/citología , Humanos , Masculino , Maxilar/citología , OsteogénesisRESUMEN
OBJECTIVE: Orthodontic treatment induces dental tissue remodeling; however, dental pulp stem cell (DPSC)-mediated pulp micro-environmental alteration is still largely uncharacterized. In the present study, we identified elevated interleukin-17A (IL17A) in the dental pulp, which induced the osteogenesis of DPSCs after orthodontic force loading. DESIGN: Tooth movement animal models were established in Sprague-Dawley rats, and samples were harvested at 1, 4, 7, 14, and 21 days after orthodontic treatment loading. DPSC self-renewal and differentiation at different time points were examined, as well as the alteration of the microenvironment of dental pulp tissue by histological analysis and the systemic serum IL17A expression level by an ELISA assay. In vitro recombinant IL17A treatment was used to confirm the effect of IL17A on the enhancement of DPSC self-renewal and differentiation. RESULTS: Orthodontic treatment altered the dental pulp microenvironment by activation of the pro-inflammatory cytokine IL17A in vivo. Orthodontic loading significantly promoted the self-renewal and differentiation of DPSCs. Inflammation and elevated IL17A secretion occurred in the dental pulp during orthodontic tooth movement. Moreover, in vitro recombinant IL17A treatment mimicked the enhancement of the self-renewal and differentiation of DPSCs. CONCLUSIONS: Orthodontic treatment enhanced the differentiation and self-renewal of DPSCs, mediated by orthodontic-induced inflammation and subsequent elevation of IL17A level in the dental pulp microenvironment.
Asunto(s)
Microambiente Celular/efectos de los fármacos , Pulpa Dental/citología , Interleucina-17/metabolismo , Ortodoncia/métodos , Células Madre/citología , Técnicas de Movimiento Dental , Animales , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Células Cultivadas , Pulpa Dental/metabolismo , Femenino , Humanos , Interleucina-17/biosíntesis , Interleucina-17/sangre , Interleucina-17/farmacología , Maxilar/citología , Modelos Animales , Osteogénesis/fisiología , Ratas , Ratas Sprague-Dawley , Células Madre/metabolismoRESUMEN
Bone graft are used in dentistry for the reconstruction of severely atrophic jaws. Fresh frozen bone has no osteogenic property but it has osteoconductive and osteoinductive properties because its matrix contains growth factors such as vascular endothelial growth factor. The purpose of the present study was to evaluate morphological and protein expression characteristics of fresh frozen bone before graft and after six months of graft in patients who needed maxillary reconstruction. After 6 month of graft we observed the presence of viable bone as evidenced by full osteocyte lacunae and by the presence of RANKR, osteocalcin positive cells and vascular endothelial growth factor. In conclusion, our findings show that the fresh frozen bone after six month of graft is for the most part viable bone, encouraging its use as an alternative to autogenous bone for reconstructing maxillary bone defects prior to implant.
Asunto(s)
Trasplante Óseo , Criopreservación , Maxilar/citología , Maxilar/metabolismo , Femenino , Humanos , MasculinoRESUMEN
The response to the mechanical loading of bone tissue has been extensively investigated; however, precisely how much strain intensity is necessary to promote bone formation remains unclear. Combination studies utilizing histomorphometric and numerical analyses were performed using the established murine maxilla loading model to clarify the threshold of mechanical strain needed to accelerate bone formation activity. For 7 days, 191 kPa loading stimulation for 30 min/day was applied to C57BL/6J mice. Two regions of interest, the AWAY region (away from the loading site) and the NEAR region (near the loading site), were determined. The inflammatory score increased in the NEAR region, but not in the AWAY region. A strain intensity map obtained from [Formula: see text] images was superimposed onto the images of the bone formation inhibitor, sclerostin-positive cell localization. The number of sclerostin-positive cells significantly decreased after mechanical loading of more than [Formula: see text] in the AWAY region, but not in the NEAR region. The mineral apposition rate, which shows the bone formation ability of osteoblasts, was accelerated at the site of surface strain intensity, namely around [Formula: see text], but not at the site of lower surface strain intensity, which was around [Formula: see text] in the AWAY region, thus suggesting the existence of a strain intensity threshold for promoting bone formation. Taken together, our data suggest that a threshold of mechanical strain intensity for the direct activation of osteoblast function and the reduction of sclerostin exists in a murine maxilla loading model in the non-inflammatory region.
Asunto(s)
Maxilar/fisiología , Modelos Biológicos , Osteoblastos/fisiología , Estrés Mecánico , Proteínas Adaptadoras Transductoras de Señales , Animales , Densidad Ósea , Recuento de Células , Glicoproteínas/metabolismo , Péptidos y Proteínas de Señalización Intercelular , Masculino , Maxilar/citología , Ratones Endogámicos C57BL , Osteocitos/citología , Osteogénesis , Soporte de PesoRESUMEN
BACKGROUND/OBJECTIVE: The aim of this study was to investigate the effects of different concentrations of ozone (O3) therapy on bone regeneration in response to an expansion of the inter-premaxillary suture in rats. MATERIALS AND METHODS: Forty-eight Wistar rats were randomly divided into four groups (n = 12). In groups I, II, and III, 1ml of O3 at 10, 25, and 40 µg/ml was injected at the premaxillary suture, respectively. In group IV (control group), 1ml of saline solution was injected at the same point during the expansion procedure for 5 days. Bone regeneration in the suture was evaluated histomorphometrically. The area of new bone and fibrotic area, the number of osteoblasts and osteoclasts, and the amount of vascularity were measured and compared. The density of the newly formed bone in the expansion area was measured by using cone beam computed tomography. Data were analyzed using the Kruskal-Wallis one-way analysis of variance and post hoc Student-Newman-Keuls tests. RESULTS: New bone area, fibrotic area, osteoblast and osteoclast numbers, and the amount of vascularity were significantly higher in experimental groups compared with the control group (P < 0.001). The density of newly formed bone (P < 0.001), new bone formation (P = 0.009), number of capillaries (P < 0.001), number of osteoclasts (P = 0.016), and number of osteoblasts (P < 0.001) in the maxillary sutures were highest in the 25 µg/ml O3 group compared with the other experimental groups and control group. CONCLUSIONS/IMPLICATIONS: The application of O3 therapy can stimulate bone regeneration in an orthopedically expanded inter-premaxillary suture during both the expansion and retention periods.
Asunto(s)
Regeneración Ósea/efectos de los fármacos , Suturas Craneales/efectos de los fármacos , Oxidantes Fotoquímicos/administración & dosificación , Ozono/administración & dosificación , Técnica de Expansión Palatina , Animales , Regeneración Ósea/fisiología , Tomografía Computarizada de Haz Cónico , Suturas Craneales/diagnóstico por imagen , Suturas Craneales/fisiología , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos/métodos , Masculino , Maxilar/citología , Maxilar/fisiología , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Osteoclastos/citología , Osteoclastos/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Osteogénesis/fisiología , Oxidantes Fotoquímicos/farmacología , Ozono/farmacología , Ratas , Ratas WistarRESUMEN
INTRODUCTION: The evaluation of bone quality at the site of the alveolar bone for a dental implant is very important. This study presents an easy technique for direct evaluation of alveolar bone quality using nondecalcified cryofilm frozen sections on human alveolar bone core samples. MATERIALS AND METHODS: Core samples harvested from alveolar bone were immediately frozen in cooled hexanen and slowly cut using a disposable tungsten carbide blade; the sliced sections were collected with adhesive cryofilms. Staining was performed using von toluidine blue and von Kossa for microscopic observations. RESULTS: All core samples clearly showed bone structure components of cortical bone, trabecular bone, bone marrow, blood vessels, and bone-related cells. CONCLUSION: These results suggest the efficacy of a nondecalcified cryofilm frozen section technique for histological observation of surgical implant sites.
Asunto(s)
Pérdida de Hueso Alveolar/patología , Crioultramicrotomía/métodos , Implantación Dental Endoósea/métodos , Mandíbula/anatomía & histología , Maxilar/anatomía & histología , Implantes Dentales , Humanos , Mandíbula/irrigación sanguínea , Mandíbula/citología , Maxilar/irrigación sanguínea , Maxilar/citología , Microscopía/métodosRESUMEN
OBJECTIVES: This study aimed to evaluate the biologic and structural phenotypes of the bone regenerated via the sandwich bone augmentation (SBA) technique, on buccal implant dehiscence defects. MATERIAL AND METHODS: Twenty-six patients with one buccal implant dehiscence defect each were randomly assigned to two groups. Both groups received a standardized amount of mineralized cancellous and cortical allogenic bone graft. In the test group, a bovine pericardium membrane was placed over the graft, while no membrane was placed in the control group. After 6 months of healing, a bone core biopsy of the regenerated bone was harvested and processed for histologic, immunohistochemical, mRNA, and micro-computed tomography (µCT) analyses. Of the 26 bone core biopsies, only six cores from the test group and six cores from the control group were suitable for the analysis. RESULTS: Bone volume (BV) in the test group was maintained, but tissue maturation appeared to be delayed. In contrast, tissue maturation appeared to be completed in the control group, but BV was compromised. Micro-CT analysis showed that specimens from the control group were more structured and mineralized compared with those from the test group. Histologic analysis showed more residual graft particles scattered in a loose fibrous connective tissue matrix with sparse bone formation in the test group, while the control group showed obvious vital bone formation surrounding the residual graft particles. Positive periostin (POSTN), sclerostin, and runt-related transcription factor-2 (RUNX2) immunoreactivities were detected in both the control and test groups. However, tartrate-resistant acid phosphatase (TRAP) positive was mostly noted in the control group. There were significant differences in POSTN, RUNX2 and VEGF expressions between the test and control groups. CONCLUSION: These findings indicated that the SBA technique was an effective method in preserving adequate structural volume while promoting new vital bone formation. Use of the collagen barrier membrane has successfully maintained the volumetric dimensions of the ridge but might have slowed down the complete maturation of the outermost layer of the grafted site.
Asunto(s)
Aumento de la Cresta Alveolar/métodos , Implantación Dental Endoósea/métodos , Histocitoquímica , Inmunohistoquímica , Maxilar/anatomía & histología , ARN/análisis , Tomografía Computarizada por Rayos X , Humanos , Maxilar/química , Maxilar/citología , Maxilar/diagnóstico por imagen , Resultado del TratamientoRESUMEN
BACKGROUND: While many studies have been performed on the characteristics and regenerative capacity of long bone periosteum, the craniofacial periosteum remains poorly understood. AIM: The aim of this study was to investigate the potential for a maxillary periosteum tunnelling procedure to induce vertical alveolar bone regeneration. MATERIALS AND METHODS: We employed a murine injury model that activates skeletal stem cells in the periosteum without overtly damaging the underlying cortical bone, preserving the integrity of the long bone and maxilla, and avoiding the introduction of pathological motion at the injury site. Further, we introduced a collagen sponge to serve as a scaffold, providing the necessary space for vertical bone regeneration. RESULTS: Periosteal elevation alone resulted in bone formation in the tibia and delayed bone resorption in the maxilla. With the presence of the collagen sponge, new bone formation occurred in the maxilla. CONCLUSIONS: Periosteal response to injury varies with anatomical location, so conclusions from long bone studies should not be extrapolated for craniofacial applications. Murine maxillary periosteum has the osteogenic potential to induce vertical alveolar bone regeneration.
Asunto(s)
Regeneración Ósea/fisiología , Maxilar/fisiología , Periostio/fisiología , Fosfatasa Ácida/análisis , Fosfatasa Alcalina/análisis , Proceso Alveolar/citología , Proceso Alveolar/fisiología , Animales , Resorción Ósea/fisiopatología , Calcificación Fisiológica/fisiología , Proliferación Celular , Colágeno/química , Células del Tejido Conectivo/citología , Etiquetado Corte-Fin in Situ , Isoenzimas/análisis , Masculino , Maxilar/citología , Maxilar/cirugía , Ratones , Modelos Animales , Osteoclastos/citología , Osteogénesis/fisiología , Periostio/citología , Periostio/cirugía , Antígeno Nuclear de Célula en Proliferación/análisis , Células Madre/fisiología , Fosfatasa Ácida Tartratorresistente , Tibia/citología , Tibia/fisiología , Tibia/cirugía , Ingeniería de Tejidos/métodos , Andamios del Tejido/químicaRESUMEN
Osteoprotegerin (OPG) is secreted by stromal and osteoblastic lineage cells and inhibits osteoclastogenesis by preventing the interaction of receptor activator of nuclear factor-κB ligand (RANKL) with receptor activator of nuclear factor-κB (RANK). In this study, the expression of OPG in osteoclasts themselves and its biological functions during osteoclastogenesis were investigated for the first time. OPG expression in vivo in the developing rat maxilla was examined by immunofluorescence analysis. OPG expression in osteoclasts during in vitro osteoclastogenesis was determined by reverse-transcription polymerase chain-reaction (RT-PCR), Western blot, and immunofluorescence staining. We determined the function of OPG produced by osteoclasts during osteoclastogenesis by silencing the OPG gene. The effects of OPG on bone-resorbing activity and apoptosis of mature osteoclasts were examined by the assay of resorptive pit formation on calcium-phosphate-coated plate and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining, respectively. In the immunofluorescence findings, strong immunoreactivities were unexpectedly seen in multinucleated tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts around the growing and erupting tooth germs in the rat alveolar bone. In vitro, OPG expression was significantly increased during the differentiation of osteoclasts from mouse bone-marrow-derived cells treated with a combination of macrophage colony-stimulating factor (M-CSF) and RANKL. Interestingly, it was found that OPG small interfering (si)RNA treatment during osteoclastogenesis enhanced the sizes of osteoclasts, but attenuated their bone-resorbing activity. Also, the increased chromosomal DNA fragmentation and caspase-3 activity in the late phase of osteoclastogenesis were found to be decreased by treatment with OPG siRNA. Furthermore, effects of OPG siRNA treatment on osteoclastogenesis and bone-resorbing activity were recovered by the treatment of exogenous OPG. These results suggest that OPG, expressed by the osteoclasts themselves, may play an auto-regulatory role in the late phase of osteoclastogenesis through the induction of apoptosis.
Asunto(s)
Osteoclastos/metabolismo , Osteoprotegerina/análisis , Fosfatasa Ácida/análisis , Proceso Alveolar/citología , Animales , Apoptosis/fisiología , Células de la Médula Ósea/efectos de los fármacos , Resorción Ósea/metabolismo , Fosfatos de Calcio/metabolismo , Diferenciación Celular/fisiología , Tamaño de la Célula , Células Cultivadas , Materiales Biocompatibles Revestidos/metabolismo , Silenciador del Gen , Homeostasis/fisiología , Isoenzimas/análisis , Factor Estimulante de Colonias de Macrófagos/farmacología , Masculino , Maxilar/citología , Ratones , Ratones Endogámicos C57BL , Osteoclastos/efectos de los fármacos , Osteoclastos/fisiología , Osteoprotegerina/genética , Ligando RANK/farmacología , Ratas , Ratas Sprague-Dawley , Fosfatasa Ácida Tartratorresistente , Erupción Dental/fisiología , Germen Dentario/citologíaRESUMEN
The functional demand imposed on bone promotes changes in the spatial properties of osteocytes as well as in their extensions uniformly distributed throughout the mineralized surface. Once spatial deformation is established, osteocytes create the need for structural adaptations that result in bone formation and resorption that happen to meet the functional demands. The endosteum and the periosteum are the effectors responsible for stimulating adaptive osteocytes in the inner and outer surfaces. Changes in shape, volume and position of the jaws as a result of skeletal correction of the maxilla and mandible require anchorage to allow bone remodeling to redefine morphology, esthetics and function as a result of spatial deformation conducted by orthodontic appliances. Examining the degree of changes in shape, volume and structural relationship of areas where mini-implants and miniplates are placed allows us to classify mini-implants as devices of subabsolute anchorage and miniplates as devices of absolute anchorage.