RESUMO
OBJECTIVE: Hyperbilirubinemia in patients with biliary atresia causes deciduous tooth injuries such as green pigmentation and dentin hypoplasia. In patients with biliary atresia who received liver transplantation, tooth structure appears to be recovered radiographically. Nevertheless, little is known about cellular mechanisms underlying bilirubin-induced damage and suppression of deciduous tooth formation. In this study, we examined the effects of bilirubin in stem cells from human exfoliated deciduous teeth (SHED) in vitro. MATERIALS AND METHODS: SHED were cultured under exposure to excess of bilirubin and then interruption of bilirubin stimulation. RESULTS: Bilirubin induced cell death and inhibited the odontogenic capacity of SHED by suppressing AKT and extracellular signal-regulated kinase 1 and 2 (ERK1/2) pathways and enhancing nuclear factor kappa B p65 (NF-κB p65) pathway. The interruption of bilirubin stimulation reduced cell death and recovered the inhibited odontogenic capacity of bilirubin-damaged SHED. The bilirubin interruption also normalized the impaired AKT, ERK1/2, and NF-κB p65 signaling pathways. CONCLUSION: These findings suggest that tooth hypodontia in patients with hyperbilirubinemia might be due to bilirubin-induced cell death and dentinogenic dysfunction of odontogenic stem cells via AKT, ERK1/2, and NF-κB pathways and also suggested that bilirubin-induced impairments in odontogenic stem cells were reversible when bilirubin stimulation is interrupted.
Assuntos
Bilirrubina/farmacologia , Morte Celular/efeitos dos fármacos , Odontogênese/efeitos dos fármacos , Células-Tronco , Dente Decíduo/citologia , Atresia Biliar/sangue , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Pré-Escolar , Humanos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Esfoliação de Dente , Fator de Transcrição RelA/metabolismoRESUMO
In orthodontic tooth movement (OTM), we should be concerned about external root resorption (ERR) as an undesirable iatrogenic problem, but its mechanisms are not fully understood. Since our previous epidemiologic studies found that patients with allergic diseases showed higher rates of ERR during orthodontic treatment, we explored the possible effect of allergic sensitization on ERR. In ovalbumin (OVA)-sensitized Brown-Norway rats, the amounts of ERR and OTM were greater than those in animals subjected to orthodontic force alone. The expression levels of RANKL and pro-inflammatory cytokines were increased in the periodontal tissues of sensitized rats with OTM, compared with control rats. Furthermore, leukotriene B4 (LTB4), a potent lipid mediator of allergic inflammation, and enzymes of the 5-lipoxygenase pathway, the biosynthetic pathway of leukotrienes, were also up-regulated. We found that low doses of aspirin suppressed ERR in allergen-sensitized rats, as well as the expressions of RANKL, pro-inflammatory cytokines, and LTB4. The present findings indicate that allergen sensitization has adverse effects on ERR under OTM, and that aspirin is a potential therapeutic agent for combating ERR.
Assuntos
Alérgenos/imunologia , Imunização , Reabsorção da Raiz/imunologia , Processo Alveolar/imunologia , Processo Alveolar/patologia , Animais , Araquidonato 5-Lipoxigenase/análise , Aspirina/farmacologia , Fenômenos Biomecânicos , Reabsorção Óssea/imunologia , Reabsorção Óssea/patologia , Inibidores de Ciclo-Oxigenase/farmacologia , Epóxido Hidrolases/análise , Doença Iatrogênica , Imunoglobulina E/sangue , Mediadores da Inflamação/análise , Interleucina-1beta/análise , Interleucina-1beta/efeitos dos fármacos , Interleucina-6/análise , Leucotrieno B4/análise , Leucotrienos/análise , Fios Ortodônticos , Ovalbumina/imunologia , Periodonto/imunologia , Ligante RANK/análise , Ligante RANK/efeitos dos fármacos , Ratos , Ratos Endogâmicos BN , Reabsorção da Raiz/prevenção & controle , Técnicas de Movimentação Dentária/efeitos adversos , Técnicas de Movimentação Dentária/instrumentação , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Regulação para CimaRESUMO
Discoveries of immunomodulatory functions in mesenchymal stem cells (MSCs) have suggested that they might have therapeutic utility in treating immune diseases. Recently, a novel MSC population was identified from dental pulp of human supernumerary teeth, and its multipotency characterized. Herein, we first examined the in vitro and in vivo immunomodulatory functions of human supernumerary tooth-derived stem cells (SNTSCs). SNTSCs suppressed not only the viability of T-cells, but also the differentiation of interleukin 17 (IL-17)-secreting helper T (Th17)-cells in in vitro co-culture experiments. In addition, systemic SNTSC transplantation ameliorated the shortened lifespan and elevated serum autoantibodies and nephritis-like renal dysfunction in systemic lupus erythematosus (SLE) model MRL/lpr mice. SNTSC transplantation also suppressed in vivo increased levels of peripheral Th17 cells and IL-17, as well as ex vivo differentiation of Th17 cells in MRL/lpr mice. Adoptive transfer experiments demonstrated that SNTSC-transplanted MRL/lpr mouse-derived T-cell-adopted immunocompromised mice showed a longer lifespan in comparison with non-transplanted MRL/lpr mouse-derived T-cell-adopted immunocompromised mice, indicating that SNTSC transplantation suppresses the hyper-immune condition of MRL/lpr mice through suppressing T-cells. Analysis of these data suggests that SNTSCs are a promising MSC source for cell-based therapy for immune diseases such as SLE.
Assuntos
Polpa Dentária/patologia , Imunoterapia/métodos , Células-Tronco Mesenquimais/imunologia , Dente Supranumerário/patologia , Transferência Adotiva/métodos , Animais , Apoptose/imunologia , Autoanticorpos/sangue , Técnicas de Cultura de Células , Diferenciação Celular/imunologia , Sobrevivência Celular/imunologia , Criança , Pré-Escolar , Técnicas de Cocultura , Feminino , Glomerulonefrite/prevenção & controle , Humanos , Imunomodulação/imunologia , Interleucina-17/imunologia , Longevidade , Lúpus Eritematoso Sistêmico/imunologia , Transplante de Células-Tronco Mesenquimais , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos MRL lpr , Camundongos Endogâmicos , Células-Tronco Multipotentes/imunologia , Linfócitos T/imunologia , Células Th17/imunologiaRESUMO
UNLABELLED: Although human orofacial bone-marrow-derived mesenchymal stem cells showed differentiation traits distinctly different from those of mesenchymal stem cells (MSCs) derived from long bone marrow (BMMSCs), mouse MSCs derived from orofacial bone have not been isolated due to technical difficulties, which in turn precludes the use of mouse models to study and cure orofacial diseases. In this study, we developed techniques to isolate and expand mouse orofacial bone/bone-marrow-derived MSCs (OMSCs) from mandibles and verified their MSC characteristics by single-colony formation, multi-lineage differentiation, and in vivo tissue regeneration. Activated T-lymphocytes impaired OMSCs via the Fas/Fas ligand pathway, as occurs in BMMSCs. Furthermore, we found that OMSCs are distinct from BMMSCs with respect to regulating T-lymphocyte survival and proliferation. Analysis of our data suggests that OMSCs are a unique population of MSCs and play an important role in systemic immunity. ABBREVIATIONS: BMMSC, bone marrow mesenchymal stem cell; HA/TCP, hydroxyapatite/tricalcium phosphate; OMSC, orofacial mesenchymal stem cell; OVX, ovariectomized.
Assuntos
Mandíbula/citologia , Células-Tronco Mesenquimais/citologia , Animais , Antígenos Ly/biossíntese , Apoptose , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Fosfatos de Cálcio , Técnicas de Cultura de Células , Diferenciação Celular , Separação Celular , Células Cultivadas , Técnicas de Cocultura , Ensaio de Unidades Formadoras de Colônias , Durapatita , Proteínas de Membrana/biossíntese , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/imunologia , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos C3H , Camundongos Nus , Fator 3 de Transcrição de Octâmero/biossíntese , Osteoblastos/citologia , Osteoblastos/metabolismo , Ovariectomia , Regeneração , Antígenos Embrionários Estágio-Específicos/biossíntese , Linfócitos T/imunologia , Alicerces TeciduaisRESUMO
OBJECTIVE: Periodontal disease is an inflammatory disorder with widespread morbidities involving both oral and systemic health. The primary goal of periodontal treatment is the regeneration of the lost or diseased periodontium. In this study, we retrospectively examined feasibility and safety of reconstructing the periodontal intrabony defects with autologous periodontal ligament progenitor (PDLP) implantation in three patients. MATERIALS AND METHODS: In this retrospective pilot study, we treated 16 teeth with at least one deep intrabony defect of probing depth (PD) > OR = 6 mm with PDLP transplantation and evaluated clinical outcome measures in terms of probing depth, gingival recession and attachment gain for a duration of 32-72 months. Furthermore, we compare PDLPs with standard PDL stem cells (PDLSCs) and confirmed that PDLPs possessed progenitor characters. RESULTS: Clinical examination indicated that transplantationof PDLPs may provide therapeutic benefit for the periodontal defects. All treated patients showed no adverse effects during the entire course of follow up. We also found that PDLPs were analogous to PDLSCs in terms of high proliferation, expression of mesenchymal surface molecules, multipotent differentiation, and in vivo tissue regain. However, PDLPs failed to express scleraxis, a marker of tendon, as seen in PDLSCs. CONCLUSIONS: This study demonstrated clinical and experimental evidences supporting a potential efficacy and safety of utilizing autologous PDL cells in the treatment of human periodontitis.
Assuntos
Perda do Osso Alveolar/cirurgia , Regeneração Óssea , Células-Tronco Multipotentes/transplante , Ligamento Periodontal/citologia , Periodontite/cirurgia , Transplante de Células-Tronco , Adulto , Animais , Células Cultivadas , Cementogênese , Durapatita , Estudos de Viabilidade , Humanos , Masculino , Camundongos , Camundongos Nus , Dente Serotino/citologia , Projetos Piloto , Estudos Retrospectivos , Alicerces TeciduaisRESUMO
OBJECTIVE: Stem cells from human exfoliated deciduous teeth (SHED) are a population of highly proliferative postnatal stem cells capable of differentiating into odontoblasts, adipocytes, neural cells, and osteo-inductive cells. To examine whether SHED-mediated bone regeneration can be utilized for therapeutic purposes, we used SHED to repair critical-size calvarial defects in immunocompromised mice. MATERIALS AND METHODS: We generated calvarial defects and transplanted SHED with hydroxyapatite/tricalcium phosphate as a carrier into the defect areas. RESULTS: SHED were able to repair the defects with substantial bone formation. Interestingly, SHED-mediated osteogenesis failed to recruit hematopoietic marrow elements that are commonly seen in bone marrow mesenchymal stem cell-generated bone. Furthermore, SHED were found to co-express mesenchymal stem cell marker, CC9/MUC18/CD146, with an array of growth factor receptors such as transforming growth factor beta receptor I and II, fibroblast growth factor receptor I and III, and vascular endothelial growth factor receptor I, implying their comprehensive differentiation potential. CONCLUSIONS: Our data indicate that SHED, derived from neural crest cells, may select unique mechanisms to exert osteogenesis. SHED might be a suitable resource for orofacial bone regeneration.
Assuntos
Regeneração Óssea/fisiologia , Células-Tronco Multipotentes/citologia , Osseointegração/fisiologia , Osteoblastos/citologia , Osteogênese/fisiologia , Transplante de Células-Tronco/métodos , Implantes Absorvíveis , Animais , Materiais Biocompatíveis , Cimentos Ósseos , Substitutos Ósseos , Fosfatos de Cálcio , Células Cultivadas , Durapatita , Humanos , Camundongos , Camundongos Endogâmicos , Células-Tronco Multipotentes/transplante , Crista Neural/citologia , Osteoblastos/fisiologia , Crânio/cirurgia , Dente Decíduo/citologiaRESUMO
UNLABELLED: Hertwig's epithelial root sheath (HERS) cells are a unique population of epithelial cells in the periodontal ligament compartment. To date, their functional role has not been fully elucidated. Our hypothesis was that HERS cells may be involved in regulating differentiation of periodontal ligament stem cells (PDLSCs) and forming cementum in vivo. In this study, we found that HERS cells may be capable of promoting PDLSC differentiation and undergoing epithelial-mesenchymal transition in vitro. Immunohistochemical staining, Western blot analysis, a transwell co-culture system, and in vivo transplantation were used to characterize the interplay between HERS cells and PDLSCs, as well as the epithelial-mesenchymal transition (EMT) of HERS cells. TGFbeta1 was capable of inducing the epithelial-mesenchymal transition of HERS cells through activating the PI3K/AKT pathway. Furthermore, HERS cells were able to form cementum-like tissue when transplanted into immunocompromised mice. ABBREVIATIONS: bone marrow mesenchymal stem cell, BMMSC; bone sialoprotein, BSP; hydroxyapatite/tricalcium phosphate, HA/TCP; Hertwig's epithelial root sheath, HERS; osteocalcin, OCN; periodontal ligament, PDL; periodontal ligament stem cell, PDLSC; phosphatidylinositol 3-kinase, PI3K.
Assuntos
Cementogênese/fisiologia , Células Epiteliais/fisiologia , Ligamento Periodontal/citologia , Adolescente , Adulto , Animais , Diferenciação Celular , Células Cultivadas , Células Epiteliais/citologia , Células Epiteliais/transplante , Humanos , Imunofenotipagem , Mesoderma/citologia , Camundongos , Camundongos Nus , Células-Tronco/fisiologia , Fator de Crescimento Transformador beta1/fisiologiaRESUMO
The craniofacial region contains many specified tissues including bone, cartilage, muscle, blood vessels and neurons. Defect or dysfunction of the craniofacial tissue after post-cancer ablative surgery, trauma, congenital malformations and progressive deforming skeletal diseases has a huge influence on the patient's life. Therefore, functional reconstruction of damaged tissues is highly expected. Bone marrow-derived mesenchymal stem cells (BMMSCs) are one of the most well characterized postnatal stem cell populations, and considered to be utilized for cell-based clinical therapies. Here, the current understanding and the potential applications in craniofacial tissue regeneration of BMMSCs are reviewed, and the current limitations and drawbacks are also discussed.
Assuntos
Células da Medula Óssea/fisiologia , Células-Tronco Mesenquimais/fisiologia , Regeneração/fisiologia , Animais , Transplante de Medula Óssea , Face/cirurgia , Humanos , Transplante de Células-Tronco Mesenquimais , Células-Tronco Multipotentes/fisiologia , Crânio/cirurgia , Engenharia TecidualRESUMO
Synovial hyperplasia is a feature of degenerative temporomandibular joint (TMJ) disease. However, the mechanism by which hyperplasia progresses in the TMJ is unknown. Based on the hypothesis that the oxidative stress generated by mechanical loading causes degenerative changes in the TMJ synovium, we investigated the generation of the highly reactive species, peroxynitrite, and the occurrence of DNA damage in the synovium. After condylar hypermobility of rat TMJs, a marker of peroxynitrite, nitrotyrosine, was localized to the nuclei and cytoplasm of the synovial lining cells and fibroblasts in synovitis-induced TMJ. DNA single-strand breaks were found in the nuclei of the synovial cells only after enzyme treatment, whereas DNA double-strand breaks were not detected. These findings indicate that condylar hypermovement induces the proliferation of synovial cells, and suggest that oxidative stress leads to the progression of synovial hyperplasia via DNA damage of the synovial cells in TMJs after mechanical loading.
Assuntos
Dano ao DNA , Instabilidade Articular/fisiopatologia , Estresse Oxidativo , Membrana Sinovial/patologia , Sinovite/fisiopatologia , Transtornos da Articulação Temporomandibular/fisiopatologia , Tirosina/análogos & derivados , Animais , Modelos Animais de Doenças , Hiperplasia , Instabilidade Articular/metabolismo , Instabilidade Articular/patologia , Masculino , Côndilo Mandibular/fisiopatologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo II , Ácido Peroxinitroso/metabolismo , Ratos , Ratos Wistar , Estresse Mecânico , Membrana Sinovial/metabolismo , Membrana Sinovial/fisiopatologia , Sinovite/metabolismo , Sinovite/patologia , Articulação Temporomandibular/metabolismo , Articulação Temporomandibular/fisiopatologia , Transtornos da Articulação Temporomandibular/metabolismo , Transtornos da Articulação Temporomandibular/patologia , Tirosina/metabolismoRESUMO
Capsaicin, the pungent substance in hot peppers, evokes a sensation of burning pain by stimulating the vanilloid receptor 1 (VR1) on primary afferent neurons. Immunohistochemistry revealed that the taste papillae in the tongue and palate are richly innervated by VR1-immunoreactive nerve fibers. Furthermore, VR1 protein expression was seen in the epithelium facing the oral cavity, although taste cells seemed to be devoid of VR1. The most conspicuous VR1 expression was observed in the epithelial cells of the palatal rugae, although there were no VR1-immunoreactive nerves there. The finding that VR1 is expressed not only in primary afferents but also in oral epithelial cells suggests that it is of great importance in the perception of capsaicin, heat, and acid in the mouth. Since VR1 is known to play a key role in nociception and inflammatory pain, it may be a new target for the treatment of oral pain.
Assuntos
Capsaicina/metabolismo , Palato/química , Receptores de Droga/biossíntese , Língua/química , Animais , Epitélio/química , Epitélio/inervação , Epitélio/metabolismo , Expressão Gênica , Técnicas Imunoenzimáticas , Masculino , Neurônios Aferentes/química , Palato/inervação , Palato/metabolismo , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Receptores de Droga/análise , Receptores de Droga/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Língua/inervação , Língua/metabolismoRESUMO
NF-kappaB plays a pivotal role in pathogenesis in general arthritis. However, the participation of NF-kappaB in inflammation of the temporomandibular joint (TMJ) is poorly understood. We examined NF-kappaB expression in rat TMJs with synovitis induced by condyle hypermobility. By immunohistochemistry, NF-kappaB immunoreactivity was found mainly in the cytoplasm, not the nucleus, of the synovial lining cells of induced-synovitis and control TMJs. Southwestern histochemistry, a new method for detecting transcription factors, showed greater NF-kappaB expression in the nucleus of the synovial lining cells in the hypertrophic synovium than in control synovium. Increased numbers of the synovial lining cells with immunoreactivity for inducible nitric oxide synthase (iNOS), which is transcriptionally regulated by NF-kappaB, were also seen in the inflamed synovium. These findings indicate that excess mechanical stress increases NF-kappaB activation in the TMJ and suggest that active NF-kappaB is involved in the progression of TMJ inflammation.
Assuntos
NF-kappa B/fisiologia , Óxido Nítrico Sintase/biossíntese , Membrana Sinovial/metabolismo , Sinovite/metabolismo , Transtornos da Articulação Temporomandibular/metabolismo , Animais , Southwestern Blotting , Ativação Enzimática , Imuno-Histoquímica , Masculino , Óxido Nítrico Sintase Tipo II , Ratos , Ratos Wistar , Estresse MecânicoRESUMO
Substance P (SP) is an important member of the tachykinin family of neuropeptides, which work as neurotransmitters or neuromodulators. Recent advances in the analysis of SP receptors, particularly the neurokinin-1 receptors (NK1-Rs) that have high affinity for SP, have demonstrated that they are distributed not only in the cells of the neuronal or immune systems but also in peripheral cells. Therefore, the effect of SP and its cellular receptors is not limited to the nervous or immune systems, but is more extensive than previously appreciated. SP-like immunoreactive (SP-LI) axons have been localized in both bone and gingival tissue, and SP receptors are widely distributed in osteoclasts, osteoblasts, and junctional epithelial cells, as well as in neutrophils and endothelial cells. The distribution of SP-LI axons and SP receptors suggests that SP may directly modulate bone metabolism and gingival tissue functions through SP receptors.
Assuntos
Osso e Ossos/metabolismo , Gengiva/metabolismo , Receptores da Neurocinina-1/fisiologia , Substância P/fisiologia , Animais , Axônios/metabolismo , Osso e Ossos/inervação , Osso e Ossos/ultraestrutura , Células Cultivadas , Gengiva/inervação , Gengiva/ultraestrutura , Imuno-Histoquímica , Microscopia Confocal , Microscopia Eletrônica , Ratos , Receptores da Neurocinina-1/análise , Substância P/análiseRESUMO
An epidemiologic examination was carried out to reveal the prevalence of the periodontal diseases and oral pigmentation in patients with Yusho in 2000. The results obtained were as follows. 1) 63 patients out of 69 patients with Yusho, who were measured periodontal pocket depth according to Ramfjord' methods, had at least one tooth with periodontal pocket deeper than 3 mm. Similarly, 188 teeth out of a total 285 examined teeth showed periodontal pocket with more than 3 mm depth. 2) In this examination, intraoral sinus tracts stoma were observed in 9 patients out of 70 patients. Radiographic examination and probing examination of pocket depth indicated that periapical lesions were involved in these intraoral sinus tract formation. 3) Oral pigmentation was observed in 46 out of 76 patients with Yusho. In this study, gingival pigmentation was most predominant among oral pigmentation. These results indicated that PCBs had yet affected the mechanism of oral pigmentation and metabolism of alveolar bone.
Assuntos
Contaminação de Alimentos , Doenças da Boca/epidemiologia , Oryza/intoxicação , Doenças Periodontais/epidemiologia , Transtornos da Pigmentação/epidemiologia , Óleos de Plantas/intoxicação , Bifenilos Policlorados/intoxicação , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Doenças da Gengiva/epidemiologia , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-IdadeRESUMO
We compared the distribution of a cysteine proteinase inhibitor, cystatin C, with that of cathepsin K in osteoclasts of the mouse tibia by immunolight and immunoelectron microscopy. Light microscopically, strong immunoreactivity for cystatin C was found extracellularly along the resorption lacuna and intracellularly in the organelles of osteoclasts. In serial sections, various patterns of cystatin C and cathepsin K localization were seen, specifically: (1) some resorption lacuna were positive for both cystatin C and cathepsin K; (2) others were positive for either cystatin C or cathepsin K, but not both; and (3) some lacuna were negative for both. In osteoclasts, the localization of cystatin C was similar to that of cathepsin K. Furthermore, cystatin C immunoreactivity was detected in preosteoclasts and osteoblasts, whereas cathepsin K was seen only in preosteoclasts. Electron microscopically, cystatin C immunoreactive products were found in the rough endoplasmic reticulum (ER), Golgi apparatus, vesicles, granules, and vacuoles of osteoclasts. These cystatin C-positive vesicles had fused or were in the process of fusion with the ampullar vacuoles (extracellular spaces) containing cystatin C-positive, fragmented, fibril-like structures. The extracellular cystatin C was deposited on and between the cytoplasmic processes of ruffled borders, and on and between type I collagen fibrils. In the basolateral region of osteoclasts, cystatin C-positive vesicles and granules also fused with vacuoles that contained cystatin C-positive or negative fibril-like structures. These results indicate that osteoclasts not only synthesize and secrete cathepsin K from the ruffled border into the bone resorption lacunae, but also a cysteine proteinase inhibitor, cystatin C. Therefore, it is suggested that cystatin C regulates the degradation of bone matrix by cathepsin K, both extracellularly and intracellularly.
Assuntos
Catepsinas/metabolismo , Cistatinas/metabolismo , Osteoclastos/metabolismo , Tíbia/metabolismo , Animais , Reabsorção Óssea/metabolismo , Catepsina K , Cistatina C , Epífises/citologia , Epífises/metabolismo , Imuno-Histoquímica , Rim/metabolismo , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Imunoeletrônica , Tíbia/citologia , Distribuição TecidualRESUMO
The distribution and fine structure of nerve fibers containing neuropeptide Y (NPY), tyrosine hydroxylase (TH), and vasoactive intestinal polypeptide (VIP) in the temporomandibular joint were investigated by both the avidin-biotin complex method and an indirect immunofluorescence technique. The innervation pattern of NPY- and TH-positive fibers differed from that of VIP-positive fibers. Specifically, the former was distributed in both the superficial and deep sublining layers, while the latter was mostly located in the deep sublining layer. NPY- and TH-immunoreactive fibers were largely confined to vascular elements; occasional fibers were observed in the synovial lining layer close to the joint cavity. More nerves with NPY and TH immunoreactivity were observed close to the upper joint compartment than near the lower compartment NPY and TH immunoreactivity was dramatically reduced in the TMJ of superior cervical ganglionectomized animals, indicating the sympathetic origin of these nerves. NPY immunoreactivity was found only in unmyelinated axons, which were located in the adventitia and adventitia-medial border of arteries or arterioles. Occasionally, axons were near the joint cavity, in areas free of vascular structures. These observations show that abundant sympathetic nerves supply the temporomandibular joint of the rat and provide a morphological basis for the involvement of different neuropeptides in vascular regulatory and modulatory functions in physiological and pathophysiological conditions.
Assuntos
Imuno-Histoquímica , Fibras Nervosas/ultraestrutura , Sistema Nervoso Simpático/ultraestrutura , Articulação Temporomandibular/inervação , Animais , Ganglionectomia , Masculino , Microscopia Imunoeletrônica , Fibras Nervosas/química , Neuropeptídeo Y/análise , Ratos , Ratos Wistar , Gânglio Cervical Superior/fisiologia , Tirosina 3-Mono-Oxigenase/análise , Peptídeo Intestinal Vasoativo/análiseRESUMO
This study demonstrated the simultaneous expression of cathepsin K (CK) mRNA by in situ hybridization and CK protein by immunoelectron microscopy in odontoclasts in mouse maxillae after experimental tooth movement. On the pressure side (the area under pressure during tooth movement), CK mRNA was detected in odontoclasts in resorption lacunae in the tooth root, in osteoclasts in bone resorption lacuane, and in fibroblasts in the periodontal ligament. Using electron microscopy, CK protein was detected at the apex of odontoclasts, intracellularly in vesicles and granules, and extracellularly in irregularly shaped vacuoles (extracellular spaces), on the plasma membrane of the ruffled border, and on and between typical striated type I collagen fibrils in the lacunae. These vesicles and granules appeared to fuse with irregular vacuoles containing CK-positive fragmented fibril-like structures close to the ruffled border. In the basolateral portion of odontoclasts, small amounts of CK-positive rough endoplasmic reticulum (ER) were found. CK-positive intracellular vacuoles (not extracellular spaces) also appeared to fuse with the vesicles and granules. However, these fused organelles rarely contained fragmented fibril-like structures. They are probably endolysosomes. The distribution of CK in odontoclasts was similar to that previously seen in osteoclasts. Furthermore, CK-positive fibril-like structures were found in the vacuoles of fibroblasts. These results indicated that during tooth movement CK is synthesized in odontoclasts on the pressure side and secreted into the tooth resorption lacunae. Therefore, CK may take part in the degradation of the dentin matrix (type I collagen fibrils and non-collagenous protein) of the tooth root, and in the subsequent intracellular degradation of endocytosed fragmented fibril-like structures in endolysosomes.
Assuntos
Catepsinas/metabolismo , Maxila/metabolismo , Osteoclastos/metabolismo , RNA Mensageiro/metabolismo , Técnicas de Movimentação Dentária/métodos , Reabsorção de Dente/metabolismo , Animais , Catepsina K , Catepsinas/imunologia , Catepsinas/ultraestrutura , Técnicas Imunoenzimáticas , Imuno-Histoquímica , Hibridização In Situ/métodos , Masculino , Maxila/citologia , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Imunoeletrônica , Sondas de Oligonucleotídeos , Osteoclastos/ultraestrutura , RNA Mensageiro/imunologia , RNA Mensageiro/ultraestrutura , Distribuição TecidualRESUMO
BACKGROUND: The role played by the internal basal lamina (IBL) and hemidesmosomes between an implant and the peri-implant epithelium (PIE) in the adherence of the epithelium to the implant is controversial. This study used rat maxilla implantation models to clarify the ultrastructure of the PIE-implant interface. METHODS: Ti-6Al-4V implants were inserted either immediately or 2 weeks after the extraction of the upper left first molar of 6- or 4-week-old rats, respectively. The junctional epithelium (JE) of the upper right molars in the same animals was used as a control. Four weeks after implantation, the animals were sacrificed to prepare specimens for light and immunoelectron microscopy. RESULTS: Under light microscopy, the PIE appeared to attach to the implant surface. Ultrastructurally, IBL, consisting of the lamina densa and lamina lucida, and hemidesmosomes were formed only in the lower region, and rarely in the middle region, of the PIE-implant interface. In control teeth, the IBL and hemidesmosomes formed throughout the dento-JE interface. Laminin-1 was found in the IBL and also in the vesicles and vacuoles of the PIE and JE cells. Statistical analysis showed that there was also a significant difference in the amount of IBL between the PIE-implant and dento-JE interfaces. CONCLUSIONS: PIE attached to the implant via hemidesmosomes and IBL in the lower region of the PIE-implant interface. Although PIE cells may secrete laminin-1, which contributes to epidermal cell adhesion, the PIE which attaches to implants only in the lower region of the interface is considered to be the poorly adhered epithelium.
Assuntos
Ligas Dentárias , Implantação Dentária Endóssea , Implantes Dentários , Gengiva/ultraestrutura , Maxila/ultraestrutura , Titânio , Ligas , Animais , Membrana Basal/ultraestrutura , Adesão Celular , Ligas Dentárias/química , Inserção Epitelial/ultraestrutura , Epitélio/ultraestrutura , Hemidesmossomos/ultraestrutura , Laminina/análise , Masculino , Microscopia Eletrônica , Microscopia Imunoeletrônica , Ratos , Ratos Wistar , Estatísticas não Paramétricas , Propriedades de Superfície , Titânio/química , Vacúolos/ultraestruturaRESUMO
The distributions of substance P (SP) and the neurokinin-1 receptor (NK1-R), the receptor preferentially activated by SP, were examined in rat gingiva by immunocytochemical methods with light and electron microscopy. SP-immunoreactive nerve fibers were located preferentially in the junctional epithelium (JE) but few in the other oral and oral sulcular epithelia. NK1-R immunoreactivity was found in the endothelial cells (capillaries and postcapillary venules underlying the JE). NK1-R-labeled and -unlabeled unmyelinated nerve fibers were located close to the blood vessels and partially or completely covered by a Schwann cell sheath. In the JE, labeled naked axons without Schwann cell sheaths were observed. Neutrophils and macrophages in the connective tissue underlying the JE and in the JE were also labeled with NK1-R. Furthermore, NK1-R was found in the JE cells. Basically, immunoreaction products for NK1-R were found throughout various cells (endothelial cells, neutrophils, and JE cells) at invaginations of the plasma membrane and in vesicular and granular structures that are probably endosomes and are found close to both the plasma membrane and the nucleus. This is a first report, demonstrating the presence of NK1-R in the gingival tissue in the normal nonstimulated condition. Furthermore, it is thought that SP may modulate the permeability of blood vessels beneath the JE, the production of antimicrobial agents in neutrophils, and the proliferation and endocytotic ability of JE cells through NK1-R.
Assuntos
Gengiva/química , Receptores da Neurocinina-1/análise , Animais , Axônios/química , Vasos Sanguíneos/química , Células Epiteliais/química , Imuno-Histoquímica , Microscopia Imunoeletrônica , Fibras Nervosas Mielinizadas/química , Ratos , Ratos Wistar , Substância P/análise , Junções Íntimas/químicaRESUMO
The localization of cathepsin K protein in mouse osteoclasts was examined by immunolight and immunoelectron microscopy using the avidin-biotin-peroxidase complex method with anti-cathepsin K (mouse) antibody. With light microscopy, a strong immunoreaction for cathepsin K was found extracellularly along the bone and cartilage resorption lacunae and detected intracellularly in vesicles, granules, and vacuoles throughout the cytoplasm of multinuclear osteoclasts and chondroclasts attached to the surface of the bone or cartilage. Mononuclear cells, probably preosteoclasts, some distance from the bone also contained a few cathepsin K-positive vesicles and granules. Cathepsin K was sometimes found in the cisternal spaces of the rough endoplasmic reticulum and vesicles of the Golgi apparatus with electron microscopy of the basolateral region of the osteoclasts. Cathepsin K-positive vesicles and granules as lysosomal compartments were present in various stages of fusion with vacuoles as endosomal compartments that contained fragmented cathepsin K-negative fibril-like structures. Some of the vacuoles (endolysosomes), which seemed to be formed by this process of fusion, contained cathepsin K-positive vesicles and fibril-like structures that did not show the regular cross striation of type I collagen fibrils. In the apical region of the osteoclasts, cathepsin K-positive vesicles and pits had already fused with or were in the process of fusing with the ampullar extracellular spaces. There were large deposits of cathepsin K on fragmented fibril-like structures without regular cross striation in the extracellular spaces, as well as on and between the cytoplasmic processes of the ruffled border. There were also extensive deposits of cathepsin K on the type I collagen fibrils with cross striation in the bone resorption lacunae. Osteoblasts and osteocytes were negative for cathepsin K. In the immunocytochemical controls, no immunoreaction was found in the osteoclasts or preosteoclasts, or on the collagen fibrils in the resorption lacunae. The results indicate that cathepsin K is produced in mature osteoclasts attached to the bone and secreted into the bone resorption lacunae. The findings suggest that cathepsin K participates in the extracellular degradation of collagen fibrils in the resorption lacunae and in the subsequent degradation of the fragmented fibrils in the endolysosomes. It is also suggested that cathepsin K degrades the organic cartilage matrix.
Assuntos
Catepsinas/análise , Fêmur/química , Osteoclastos/química , Osteócitos/química , Animais , Catepsina K , Fêmur/ultraestrutura , Técnicas Imunoenzimáticas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Imunoeletrônica , Osteoclastos/ultraestrutura , Osteócitos/ultraestruturaRESUMO
Substance P (SP) is a neuropeptide that is released from axons of sensory neurons and causes signal transduction through the activation of the neurokinin-1 receptor (NK1-R). The present study demonstrates the distribution of SP-like-immunoreactive (SP-LI) axons and the localization of NK1-Rs in rat bone tissue using the avidin-biotin-peroxidase complex method. Axons with SP-LI were commonly found near the trabecular bone in the temporal bone marrow, but they were only sparsely distributed in the mandible, femur, and tibia. Immunoreactivity for NK1-Rs was found on the plasma membrane and in the cytoplasm of the osteoclasts. In the osteoblasts and osteocytes, a small number of weak, punctate immunoreactive products of NK1-Rs were distributed close to the plasma membrane. At the electron-microscopic level, immunoreactivity for NK1-R was distributed mainly in the whole cytoplasm, except for the clear zone of the osteoclasts, and in pit-like structures along the plasma membrane. The NK1-R-immunoreactive structures in the cytoplasm were divided into two types of organelles, consisting of vesicular and vacuolar structures (probably transport vesicles and early endosomes). In the osteoblasts and osteocytes, the number of NK1-R-positive vesicular structures was fewer than in the osteoclasts. These results thus suggest that SP secreted by the sensory axons could directly modulate bone metabolism via NK1-Rs.