RESUMO
AIM: This study evaluates the early volumetric changes after buccal soft tissue contour augmentation around implants with a porcine collagen matrix (CM) vs. the subepithelial connective tissue graft (SCTG) from the palate. MATERIALS AND METHODS: 14 patients were enrolled after early implant placement with simultaneous contour augmentation and persistent buccal tissue deficits. At implant exposure, buccal soft tissues were thickened with the CM (n = 7) or the SCTG (n = 7). Impressions were taken before and after surgery, after ten days, one, three and six months. Impressions were digitized and augmented regions 3D evaluated (soft tissue volume (mm3 , %)/thickness (mm)). RESULTS: Volume increase (mm3 ) after 6 months was 19.56 ± 8.95 mm3 (CM) and 61.75 ± 52.69 mm3 (SCTG) (insignificant, p = .058). In percentage, this was a volume loss of the initially augmented soft tissue volume (100%) of 81.76% in the CM group and 56.39% in the SCTG group (6 months). The mean soft tissue thickness increase (mm) in the buccal contour after 6 months was 0.30 ± 0.16 mm (CM) and 0.80 ± 0.61 mm (SCTG) (insignificant, p = .071). CONCLUSION: The early healing phase is associated with a significant volume loss of the soft tissues. The SCTG shows insignificant superiority compared to the CM.
Assuntos
Implantes Dentários , Gengiva , Animais , Autoenxertos , Colágeno , Tecido Conjuntivo , Humanos , SuínosRESUMO
OBJECTIVES: To evaluate whether zirconia implants demonstrate differences in hard and soft tissue integration compared to titanium implants in preclinical studies. MATERIAL AND METHODS: In March 2017, electronic (MEDLINE, EMBASE) and hand search was performed to identify preclinical studies comparing zirconia and titanium implants. Primary outcomes were bone-to-implant contact (BIC) and removal torque out (RTQ), respectively, push-in (PI) measurements. Secondary outcomes included biologic width (BW) dimensions. RESULTS: A total of 37 studies were included for data extraction after screening of 91 from 1,231 selected titles. Thirty-seven experimental studies using six different species were identified. The follow-up periods ranged between 0.4 and 56 weeks. For titanium, mean values of 59.1% (95% CI: 53.3 - 64.8), 102.6 Ncm (95% CI: 81.5 - 123.6), and 25.1 N (95% CI: 20.2 - 30.0) for BIC, RTQ, and PI were estimated, respectively. The mean values for zirconia were 55.9% (95% CI: 51.6 - 60.1), 71.5 Ncm (95% CI: 51.1 - 91.9), and 22.0 N (95% CI: 13.2 - 30.7) for corresponding parameters. Confounding factors such as animal species, implant material, loading protocol, and study or loading duration significantly influenced the outcomes. Similar qualitative soft tissue integration was reported for zirconia and titanium implants. However, faster maturation processes of epithelial and connective tissues around zirconia implants were assumed. Quantitatively, similar BW dimensions were evaluated for titanium (3.5 mm; 95% CI: 2.9 - 4.2) and zirconia (3.2 mm; 95% CI: 2.7 - 3.7), whereas the loading protocol significantly influenced the outcomes. CONCLUSIONS: Zirconia and titanium implants demonstrate a similar soft and hard tissue integration capacity. However, titanium tended to show a faster initial osseointegration process compared to zirconia. Importantly, not only material characteristics but predominantly animal species and study protocols can significantly influence the outcomes.
Assuntos
Implantes Dentários , Animais , Planejamento de Prótese Dentária , Osseointegração , Propriedades de Superfície , Titânio , ZircônioRESUMO
OBJECTIVES: To evaluate implant survival, peri-implant marginal bone loss, technical, and biological complications as well as aesthetic outcomes of zirconia implants in clinical studies. MATERIAL AND METHODS: Electronic (Medline, Embase) and hand searches were performed to identify clinical studies published between January 2004 and March 2017 investigating zirconia dental implants with a mean follow-up of at least 12 months. Primary outcomes were implant survival and peri-implant marginal bone loss. Secondary outcomes included technical and biological complications as well as aesthetic outcomes. Meta-analyses were performed to estimate implant survival and marginal bone loss. RESULTS: From 943 titles, 264 abstracts were selected. Subsequently, 80 full-text articles were screened, and 18 studies were included for data extraction. One- (14 studies) and 2-piece zirconia implants (4 studies) were investigated. Commercially available (CA) (510 implants, 398 patients) and not commercially available (NCA) zirconia implants (618 implants, 343 patients) were identified. For CA implants (follow-up: 12-61.20 months), technical complications (1.6%), implant fractures (0.2%) and biological complications (4.2%) were reported. Meta-analyses estimated 1- and 2-year survival rates of 98.3% (95% CI: 97.0%-99.6%) and 97.2% (95% CI: 94.7%-99.7%), respectively, and a mean 1-year marginal bone loss of 0.7 mm (95% CI: 0.4-1.0 mm). CONCLUSIONS: Since 2004, the survival rates of CA implants significantly improved compared with NCA implants. CA 1-piece zirconia implants showed similar 1- and 2-year mean survival rates and marginal bone loss after 1 year compared with published data for titanium implants. However, more clinical long-term data are needed to confirm the presently evaluated promising short-term outcomes.
Assuntos
Implantes Dentários , Materiais Dentários/química , Planejamento de Prótese Dentária , Ítrio/química , Zircônio/química , Perda do Osso Alveolar , Bases de Dados Factuais , Prótese Dentária Fixada por Implante , Falha de Restauração Dentária , Prótese Parcial Fixa , Humanos , Análise de Sobrevida , Resultado do TratamentoRESUMO
AIM: This study evaluates a porcine collagen matrix (CM) for soft tissue thickening in comparison to the subepithelial connective tissue graft (SCTG). MATERIAL AND METHODS: In eight beagle dogs, soft tissue thickening was performed at the buccal aspects of the upper canines (SCTG and CM). Impressions were taken before augmentation (i1), after surgery (i2), after one (i3), three (i4) and ten month (i5). Casts were optically scanned with a 3D scanner and each augmented region (unit of analysis) evaluated (primary outcome variable: volume increase in mm(3) ; secondary outcome variables: volume increase in percent, mean and maximum thickness increases in mm). RESULTS: 3D tissue measurements after surgery revealed a significant higher volume increase in the CM (86.37 mm(3) ± 35.16 mm(3) ) than in the SCTG group (47.65 mm(3) ± 17.90 mm(3) ). After 10 months, volume increase was non-significant between groups (SCTG:11.36 mm(3) ± 9.26 mm(3) ; CM: 8.67 mm(3) ± 13.67 mm(3) ). Maximum soft tissue thickness increase (i1-i5) was 0.66 mm ± 0.29 mm (SCTG) and 0.79 mm ± 0.37 mm (CM) with no significant difference. CONCLUSIONS: Ten months after soft tissue thickening, the CM is statistically non-inferior to the SCTG in terms of soft tissue volume and thickness increase. Further 3D studies are needed to confirm the data.
Assuntos
Tecido Conjuntivo , Animais , Colágeno , Cães , Gengiva , Retração Gengival , Suínos , Raiz DentáriaRESUMO
We present a mild one-pot freeze gelation process for fabricating near-net, complex-shaped hydroxyapatite scaffolds and to directly incorporate active proteins during scaffold processing. In particular, the direct protein incorporation enables a simultaneous adjustment and control of scaffold microstructure, porosity, resorbability and enhancement of initial mechanical and handling stability. Two proteins, serum albumin and lysozyme, are selected and their effect on scaffold stability and microstructure investigated by biaxial strength tests, electron microscopy, and mercury intrusion porosimetry. The resulting hydroxyapatite/protein composites feature adjustable porosities from 50% to 70% and a mechanical strength ranging from 2 to 6 MPa comparable to that of human spongiosa without any sintering step. Scaffold degradation behaviour and protein release are assessed by in vitro studies. A preliminary in vivo assessment of scaffold biocompatibility and resorption behaviour in adult domestic pigs is discussed. After implantation, composites were resorbed up to 50% after only 4 weeks and up to 65% after 8 weeks. In addition, 14% new bone formation after 4 weeks and 37% after 8 weeks were detected. All these investigations demonstrate the outstanding suitability of the one-pot-process to create, in a customisable and reliable way, biocompatible scaffolds with sufficient mechanical strength for handling and surgical insertion, and for potential use as biodegradable bone substitutes and versatile platform for local drug delivery.
Assuntos
Materiais Biocompatíveis/química , Substitutos Ósseos/química , Durapatita/química , Nanocompostos/química , Proteínas/química , Implantes Absorvíveis , Animais , Materiais Biocompatíveis/farmacocinética , Materiais Biocompatíveis/farmacologia , Regeneração Óssea/efeitos dos fármacos , Substitutos Ósseos/farmacocinética , Substitutos Ósseos/farmacologia , Bovinos , Galinhas , Durapatita/farmacocinética , Durapatita/farmacologia , Feminino , Teste de Materiais , Muramidase , Proteínas/farmacocinética , Proteínas/farmacologia , Soroalbumina Bovina , SuínosRESUMO
OBJECTIVE: The objective of this study was to investigate if osseous regeneration can be accelerated by involvement of periosteal tissue. Bone defect regeneration could be accelerated by the involvement of periosteal tissue if osteogenic cell signalling is maintained within the defect. It was questioned if local cell-mediated BMP-2 gene delivery makes a cell occlusive membrane dispensable during bone critical size defect regeneration. METHODS: PEG matrix (degradation time 10 days) and PEG membrane (degradation time 120 days) were used in the pig calvarial model. Cylindrical (1 × 1 cm) critical size defects (CSD) (9 per animal; 20 animals) were filled with: (i) particulated autologous bone, covered with PEG membrane (group 1); (ii) HA/TCP, covered with PEG membrane (group 2); (iii) HA/TCP, mixed with PEG matrix (group 3); and (iv) HA/TCP mixed with BMP-2-transfected osteoblasts and PEG matrix (group 4). BMP-2/4 gene transfer: liposomal in vitro transfection of BMP-2/V5-tag fusion-protein. Quantitative histomorphometry (toluidine blue staining) after 2, 4 and 12 weeks assessed bone formation. Semiquantitative immunohistochemistry estimated the expression of BMP-2, V5-tag, Runx-2 and Sox9. RESULTS: PEG matrix embedded BMP-2 expressing cells presented higher bone formation (P < 0.05) than HA/TCP + PEG matrix defect filling or PEG membrane covering (HA/TCP filling) after 12 weeks. Highest expression of BMP-2, Runx-2 and lowest expression of fibrous tissue marker Sox9 was seen in the BMP-2 group. CONCLUSION: PEG matrix embedded BMP-2 expressing cells are capable to maintain osteogenic signalling and to accelerate osseous defect regeneration in absence of a cell occlusive membrane.
Assuntos
Doenças Ósseas/cirurgia , Proteína Morfogenética Óssea 2/uso terapêutico , Regeneração Óssea/fisiologia , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Membranas Artificiais , Osteogênese/fisiologia , Crânio/cirurgia , Alicerces Teciduais/química , Implantes Absorvíveis , Animais , Autoenxertos/transplante , Proteína Morfogenética Óssea 2/genética , Substitutos Ósseos/uso terapêutico , Transplante Ósseo/métodos , Linhagem Celular , Subunidade alfa 1 de Fator de Ligação ao Core/análise , Modelos Animais de Doenças , Técnicas de Transferência de Genes , Regeneração Tecidual Guiada/métodos , Humanos , Hidroxiapatitas/uso terapêutico , Osteoblastos/fisiologia , Periósteo/fisiologia , Distribuição Aleatória , Proteínas Recombinantes de Fusão/análise , Fatores de Transcrição SOX9/análise , Suínos , Fatores de Tempo , Transfecção/métodosRESUMO
BACKGROUND: A free gingival graft (FGG) is currently the gold standard for augmenting small areas of keratinized mucosa. The porcine collagen matrix (CM) represents an alternative to autologous tissue harvesting. This study aims to compare the CM versus FGGs for augmenting keratinized peri-implant mucosa based on clinical and histologic evaluations. METHODS: The study included 14 patients who underwent a vestibuloplasty with either a FGG from the palate (n = 7) or the CM (n = 7). An implant-fixed vestibular retention splint was inserted for 30 days. Follow-up examinations were performed at 4, 10, 30, and 90 days after surgery. Width of keratinized mucosa was measured in the region of each implant (days 10, 30, and 90). After 90 days, a biopsy was harvested for histologic and immunohistologic analyses. To characterize newly formed soft tissue, the authors stained for tissue-and differentiation-specific markers, cytokeratin (CK) 5/6, 13, and 14, to detect presence or absence of keratinization. RESULTS: The groups showed similar healing, with increased peri-implant keratinized mucosa. The CM group had overall significantly shorter operation times than the FGG group. Both groups showed similar overall shrinkage (32.98% CM versus 28.35% FGG). All biopsies showed a multilayered, keratinized, squamous epithelium. CKs 5/6 and 14 were detected in the basal and suprabasal layers, and spots of CK 13 were detected in the suprabasal layer. CONCLUSIONS: During the whole observation period, both groups showed comparable clinical and histologic outcomes. Within the limitations of the present study, CM seems to be a promising alternative for the regeneration of keratinized mucosa without tissue harvesting. Comparative long-term studies are needed to investigate changes over time.
Assuntos
Colágeno/uso terapêutico , Gengiva/transplante , Vestibuloplastia/métodos , Adulto , Idoso , Animais , Biomarcadores/análise , Biópsia/métodos , Implantes Dentários , Epitélio/patologia , Feminino , Seguimentos , Gengiva/patologia , Humanos , Queratina-13/análise , Queratina-14/análise , Queratina-5/análise , Queratina-6/análise , Queratinas , Masculino , Pessoa de Meia-Idade , Duração da Cirurgia , Contenções , Suínos , Resultado do Tratamento , Cicatrização/fisiologiaRESUMO
PURPOSE: This study addressed the suitability of a polyethylene glycol (PEG) matrix as scaffold for cell-mediated local BMP-2 gene transfer in a calvarial critical size defect (CSD) model. MATERIALS AND METHODS: PEG matrix (degradation time 10 days) and PEG membrane (degradation time 120 days) were used in the pig calvarial model. Cylindrical (1 × 1 cm) CSD (9 per animal; 20 animals) were filled with: (i) HA/TCP, covered by PEG membrane (group 1); (ii) HA/TCP, mixed with PEG matrix (group 2); and (iii) HA/TCP mixed with BMP-2 transfected osteoblasts and PEG matrix (group 3). BMP-2/4 gene transfer: liposomal in vitro transfection of BMP-2/V5-tag fusion-protein. Quantitative histomorphometry (toluidine blue staining) after 2, 4 and 12 weeks assessed bone formation. Semiquantitative immunohistochemistry estimated the expression of BMP-2 and V5-tag. RESULTS: Group 3 showed significantly higher new bone formation than groups 1, 2 at 4 (P < 0.05) and 12 (P < 0.02) weeks. BMP-2-V5-tag was detected for 4 weeks. BMP-2 expression in group 3 was higher compared to all other groups after 2 and 4 (P < 0.02) weeks. CONCLUSIONS: The PEG matrix serves as scaffold for cell-mediated BMP-2 gene delivery in guided bone regeneration facilitating cell survival and protein synthesis for at least 4 weeks. Local BMP-2 gene delivery by PEG matrix-embedded cells leads to increased bone formation during critical size defect regeneration.
Assuntos
Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 2/farmacologia , Regeneração Óssea/efeitos dos fármacos , Técnicas de Transferência de Genes , Membranas Artificiais , Polietilenoglicóis/farmacologia , Crânio/cirurgia , Alicerces Teciduais , Animais , Imuno-Histoquímica , Lipossomos , Suínos , Cloreto de TolônioRESUMO
BACKGROUND: Bisphosphonate associated osteonecrosis of the jaw (BRONJ) implies an impairment in oral hard- and soft tissue repair. An understanding of the signal transduction alterations involved can inform therapeutic strategies. Transforming growth factor ß1 (TGFß1) is a critical regulator of tissue repair; galectin-3 mediates tissue differentiation and specifically modulates periodontopathic bacterial infection. The aim of this study was to compare the expression of TGFß1-related signaling molecules and Galectin-3 in BRONJ-affected and healthy mucosal tissues. To discriminate between BRONJ-specific impairments in TGFß1 signaling and secondary inflammatory changes, the results were compared to the expression of TGFß1 and Galectin-3 in mucosal tissues with osteoradionecrosis. METHODS: Oral mucosal tissue samples with histologically-confirmed BRONJ (n = 20), osteoradionecrosis (n = 20), and no lesions (normal, n = 20) were processed for immunohistochemistry. Automated staining with an alkaline phosphatase-anti-alkaline phosphatase kit was used to detect TGFß1, Smad-2/3, Smad-7, and Galectin-3. We semiquantitatively assessed the ratio of stained cells/total number of cells (labeling index, Bonferroni-adjustment). RESULTS: TGFß1 and Smad-2/3 were significantly decreased (p < 0.032 and p(0.028, respectively) in the BRONJ samples and significantly increased (p < 0.04 and p <0.043, respectively) in the osteoradionecrosis samples compared to normal tissue. Smad-7 was significantly increased (p < 0.031) in the BRONJ group and significantly decreased (p < 0.026) in the osteoradionecrosis group. Galectin-3 staining was significantly (p < 0.025) increased in both the BRONJ and the osteoradionecrosis (p < 0.038) groups compared to the normal tissue group. However, Galectin-3 expression was significantly higher in the BRONJ samples than in the osteoradionecrosis samples (p < 0.044). CONCLUSION: Our results showed that disrupted TGFß1 signaling was associated with delayed periodontal repair in BRONJ samples. The findings also indicated that impairments in TGFß1-signaling were different in BRONJ compared to osteoradionecrosis. BRONJ appeared to be associated with increased terminal osseous differentiation and decreased soft tissue proliferation. The increase in Galectin-3 reflected the increase in osseous differentiation of mucoperiosteal progenitors, and this might explain the inflammatory anergy observed in BRONJ-affected soft tissues. The results substantiated the clinical success of treating BRONJ with sequestrectomy, followed by strict mucosa closure. BRONJ can be further elucidated by investigating the specific intraoral osteoimmunologic status.
Assuntos
Difosfonatos/efeitos adversos , Galectina 3/metabolismo , Doenças Maxilomandibulares/induzido quimicamente , Arcada Osseodentária/patologia , Osteonecrose/induzido quimicamente , Transdução de Sinais , Fator de Crescimento Transformador beta1/metabolismo , Biópsia , Humanos , Imuno-Histoquímica , Doenças Maxilomandibulares/patologia , Osteonecrose/patologia , Periósteo/metabolismo , Periósteo/patologia , Proteínas Smad/metabolismo , Regulação para CimaRESUMO
OBJECTIVES: Impaired vascularization in the etiopathology of aminobisphosphonate-associated osteonecrosis of the jaw (BONJ) is assumed, but evidence is lacking. This immunohistochemical study differentiated vascularization and angiogenesis in BONJ-adjacent mucoperiosteal tissue. STUDY DESIGN: Twenty BONJ (after zoledronate treatment) and 20 control mucoperiosteal tissue samples were processed with an autostaining-based alkaline phosphatase-antialkaline phosphatase staining kit. Vascularization was assessed by CD31 staining and angiogenesis-related neovessels by CD105 staining. The ratio of stained capillary area to total area of visible field was assessed. Statistics included Bonferroni adjustment. RESULTS: CD31-stained microvessels were detected in each section and CD105-stained neovessels in each control. BONJ-adjacent mucoperiosteal tissue showed significantly fewer CD105-positive vessels in capillary areas (P < .05) than control samples. CD31-stained capillary area was not significantly reduced in mucoperiosteal BONJ-samples. CONCLUSIONS: Angiogenesis is impaired in BONJ-related mucoperiosteal tissue, but vascularization remains unaffected. Vessel remodeling and neovessel formation is delayed in BONJ, resulting in impaired tissue regeneration of bisphosphonate-exposed oral mucosa.
Assuntos
Processo Alveolar/irrigação sanguínea , Conservadores da Densidade Óssea/efeitos adversos , Difosfonatos/efeitos adversos , Doenças Maxilomandibulares/induzido quimicamente , Neovascularização Fisiológica/efeitos dos fármacos , Osteonecrose/induzido quimicamente , Fosfatase Alcalina/análise , Processo Alveolar/efeitos dos fármacos , Antígenos CD/análise , Capilares/efeitos dos fármacos , Capilares/patologia , Corantes , Tecido Conjuntivo/irrigação sanguínea , Tecido Conjuntivo/efeitos dos fármacos , Endoglina , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/patologia , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/patologia , Humanos , Processamento de Imagem Assistida por Computador/métodos , Imidazóis/efeitos adversos , Imuno-Histoquímica , Doenças Maxilomandibulares/patologia , Mucosa Bucal/irrigação sanguínea , Mucosa Bucal/efeitos dos fármacos , Osteonecrose/patologia , Periósteo/irrigação sanguínea , Periósteo/efeitos dos fármacos , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , Receptores de Superfície Celular/análise , Ácido ZoledrônicoRESUMO
BACKGROUND: Bone-destructive disease treatments include bisphosphonates and antibodies against the osteoclast differentiator, RANKL (aRANKL); however, osteonecrosis of the jaw (ONJ) is a frequent side-effect. Current models fail to explain the restriction of bisphosphonate (BP)-related and denosumab (anti-RANKL antibody)-related ONJ to jaws. Msx-1 is exclusively expressed in craniofacial structures and pivotal to cranial neural crest (CNC)-derived periodontal tissue remodeling. We hypothesised that Msx-1 expression might be impaired in bisphosphonate-related ONJ. The study aim was to elucidate Msx-1 and RANKL-associated signal transduction (BMP-2/4, RANKL) in ONJ-altered and healthy periodontal tissue. METHODS: Twenty ONJ and twenty non-BP exposed periodontal samples were processed for RT-PCR and immunohistochemistry. An automated staining-based alkaline phosphatase-anti-alkaline phosphatase method was used to measure the stained cells:total cell-number ratio (labelling index, Bonferroni adjustment). Real-time RT-PCR was performed on ONJ-affected and healthy jaw periodontal samples (n = 20 each) to quantitatively compare Msx-1, BMP-2, RANKL, and GAPDH mRNA levels. RESULTS: Semi-quantitative assessment of the ratio of stained cells showed decreased Msx-1 and RANKL and increased BMP-2/4 (all p < 0.05) expression in ONJ-adjacent periodontal tissue. ONJ tissue also exhibited decreased relative gene expression for Msx-1 (p < 0.03) and RANKL (p < 0.03) and increased BMP-2/4 expression (p < 0.02) compared to control. CONCLUSIONS: These results explain the sclerotic and osteopetrotic changes of periodontal tissue following BP application and substantiate clinical findings of BP-related impaired remodeling specific to periodontal tissue. RANKL suppression substantiated the clinical finding of impaired bone remodelling in BP- and aRANKL-induced ONJ-affected bone structures. Msx-1 suppression in ONJ-adjacent periodontal tissue suggested a bisphosphonate-related impairment in cellular differentiation that occurred exclusively jaw remodelling. Further research on developmental biology-related unique features of jaw bone structures will help to elucidate pathologies restricted to maxillofacial tissue.
Assuntos
Difosfonatos/efeitos adversos , Doenças Maxilomandibulares/induzido quimicamente , Fator de Transcrição MSX1/metabolismo , Osteonecrose/induzido quimicamente , Proteínas Morfogenéticas Ósseas/genética , Gliceraldeído-3-Fosfato Desidrogenases/genética , Humanos , Imuno-Histoquímica , Doenças Maxilomandibulares/metabolismo , Fator de Transcrição MSX1/genética , Osteonecrose/metabolismo , Ligante RANK/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND: Collagen membranes have been developed to overcome the problem of limited availability of skin grafts. Vascularization and restricted functional epithelization limit the success of bioartificial constructs. OBJECTIVE: To compare the vascularization, epithelization, and integration of a porcine collagen I/III membrane with that of split-thickness skin grafts on skin wounds. MATERIALS AND METHODS: In 21 adult pigs, full-thickness skin defects on the rear side of the ear healed by split-thickness skin grafting, by covering with the membrane, or by free granulation. Skin samples on postoperative days 1, 3, 7, 14, 21, and 28 were evaluated histologically (hematoxylin-eosin, Sirius Red) and using immunohistochemistry (cytokeratin 5/6, transforming growth factor beta receptor (TGFbetaR-III) and immunoblot (TGFbeta(1,3), Smad2/3). Epithelial thickness and TGFbetaR-III-positive capillary area were quantitatively assessed. RESULTS: Epithelization and vascularization in the membrane group were not significantly different from in the group treated with a split-thickness skin graft. Free granulation showed significantly slower epithelization and vascularization (p<.05). TGFbeta(1) and Smad2/3 complex expression were high during free granulation. Matrix was distinguishable until day 7. CONCLUSIONS: This membrane serves as a suitable full-thickness dermal substitute, because the membrane is vascularized faster than free granulation tissue and enables early epithelization.
Assuntos
Colágeno Tipo III/uso terapêutico , Colágeno Tipo I/uso terapêutico , Derme/lesões , Alicerces Teciduais , Cicatrização/fisiologia , Ferimentos Penetrantes/terapia , Animais , Derme/irrigação sanguínea , Derme/patologia , Neovascularização Fisiológica/fisiologia , Pele Artificial , Proteínas Smad Reguladas por Receptor/metabolismo , Suínos , Fator de Crescimento Transformador beta/metabolismo , Ferimentos Penetrantes/metabolismo , Ferimentos Penetrantes/patologiaRESUMO
Highly porous titanium structures are widely used for maxillofacial and orthopedic surgery because of their excellent mechanical properties similar to those of human bone and their facilitation of bone ingrowth. In contrast to common methods, the generation of porous titaniumproducts by selective electron beam melting (SEBM), an additive manufacturing technology, overcomes difficulties concerning the extreme chemical affinity of liquid titanium to atmospheric gases which consequently leads to strongly reduced ductility of the metal. The purpose of this study was to assess the suitability of a smooth compact and a porous Ti-6Al-4V structure directly produced by the SEBM process as scaffolds for bone formation. SEBM-processed titanium implants were placed into defects in the frontal skull of 15 domestic pigs. To evaluate the direct contact between bone and implant surfaces and to assess the ingrowth of osseous tissue into the porous structure, microradiographs and histomorphometric analyses were performed 14, 30, and 60 days after surgery. Bone ingrowth increased significantly during the period of this study. After 14 days the most outer regions of the implants were already filled with newly formed bone tissue (around 14%). After 30 days the bone volume inside the implants reached almost 30% and after 60 days abundant bone formation inside the implants attained 46%. During the study only scarce bone-implant contact was found around all implants, which did not exceed 9% around compact specimens and 6% around porous specimens after 60 days. This work demonstrates that highly porous titanium implants with excellent interconnectivity manufactured using the SEBM method are suitable scaffolds for bone ingrowth. This technique is a good candidate for orthopedic and maxillofacial applications.
Assuntos
Elétrons , Teste de Materiais/métodos , Titânio/farmacologia , Ligas , Animais , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/patologia , Microscopia Eletrônica de Varredura , Tamanho do Órgão/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Porosidade/efeitos dos fármacos , Próteses e Implantes , Coloração e Rotulagem , Sus scrofa , Cloreto de Tolônio/metabolismoRESUMO
Layer-by-layer self-assembled films of molecular oligoelectrolytes were used to modify Ti-6Al-4V surfaces in order to test their ability as potential drug delivery system. With regard to medical application the in vitro behavior of the modified material was investigated. The Ti-6Al-4V (6% aluminium, 4% vanadium) material was treated in a layer-by-layer (LbL) process with 2, 4, 6 and 8 layers of molecular oligoelectrolytes 1 and 2 and thereby doped with a fluorescent reporter molecule 2. Human osteoblasts were cultured for a period up to 5 days on the modified material. Ti-6Al-4V surfaces without modification were used as control. In order to investigate the in vitro behavior of the coating as well as the influence of components of the coating on osteoblastic cells, respectively, cell proliferation, differentiation and attachment of hFOB cells were observed by means of cell number, osteoblastic gene expression and fluorescence microscopy. Degradation behavior of the OEM (oligoelectrolyte multilayer film) was examined using optical spectroscopy. Measurement data imply that the layer-by-layer coating was successfully assembled on the Ti surface and endures steam sterilization. The fluorescence signal in cell culture medium increased strictly linear with increasing pre-assembled number of layers on the surface. Proliferation rates of the cells in experimental groups did not differ significantly from each other (P >or= 0.783). Differentiation pattern was not significantly changed by the coating. The fluorescent reporter component of the film was absorbed by osteoblastic cells and was detected by fluorescence microscopy.
Assuntos
Eletrólitos/química , Titânio/química , Ligas , Sequência de Bases , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Primers do DNA , Humanos , Técnicas In Vitro , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Osteoblastos/citologia , Reação em Cadeia da Polimerase , Propriedades de SuperfícieRESUMO
BACKGROUND: Increased levels of interleukin-10 (IL-10) have been observed in patients with oral cancer, possibly as a result of suppression of the immune response. Based on this, the -1082A/G polymorphism, which influences IL-10 gene expression level, was investigated in regard to its possible association with risk for oral cancer. PATIENTS AND METHODS: The polymorphism was examined in DNA samples of 144 patients with oral squamous cell carcinoma and 141 healthy controls of equivalent gender, age and ethnicity. RESULTS: The detected allele frequencies for the high expression G allele were significantly higher in patients compared to controls (34.7% versus 21.3%, respectively, p=0.0004), as well as in patients that were smokers but not those that were heavy alcohol consumers. This highly significant difference in G allele frequency was mainly due to the increase of AG heterozygotes in patients compared to controls (OR 3.05, 95% CI 1.84-5.05). CONCLUSION: These findings suggest that the high expression G allele of the -1082A/G polymorphism of the inflammation and angiogenesis-related IL-10 is strongly associated with increased risk for oral cancer.
Assuntos
Carcinoma de Células Escamosas/genética , Interleucina-10/genética , Neoplasias Bucais/genética , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo ÚnicoRESUMO
The aim of the present study was to evaluate the suitability of cellulose-based scaffolds coated with pure sodium silicate gel and sodium silicate gels accumulated with different concentrations of the bisphosphonate pamidronate as scaffolds for attachment, proliferation and differentiation of human fetal osteoblasts (hFOB 1.19). Human osteoblasts were cultured in vitro for a period up to 14 days on different cellulose scaffolds. Unmodified and sodium silicate coated cellulose scaffolds were used as control. Two surface-coated modifications of cellulose were applied. The scaffolds were coated in a modified two-step dip coating process with pure sodium silicate gel and pamidronate enriched sodium silicate gel, respectively. In order to investigate the influence of the pamidronate, concentrations of 0.667 mg Na-pamidronate/ml sodium silicate solution, 0.333 mg Na-pamidronate/ml sodium silicate solution and 3.33 x 10(-3) mg Na-pamidronate/ml sodium silicate solution were used for the coating process. Cell proliferation, vitality and attachment were examined by means of cell counting, WST-1 test, fluorescence and scanning electron microscopy. The relative grade of differentiation of hFOB cells was examined by using quantitative real-time polymerase chain reaction (qRT-PCR) analysis for the gene expression of alkaline phosphatase and osteocalcin. Proliferation and differentiation of human osteoblasts was enhanced by the sodium silicate coatings accumulated with pamidronate compared to pure sodium silicate coatings. There was a reciprocal correlation of vitality with the concentration of pamidronate. The highest vitality was found on surfaces with the lowest pamidronate accumulation. Alkaline phosphatase, an early differentiation marker, was overexpressed after 7 days in cells on all pamidronate-containing surfaces (up to 350% compared to untreated cellulose). Osteocalcin, a late differentiation marker, was overexpressed after 14 days in cells on all coated surfaces (up to 300,000% compared to untreated cellulose). The results indicate that due to the modified coating procedure a homogeneous coating and thus, an enhancement of cell attachment and subsequent cellular functions can be achieved. Low concentrations of pamidronate seem to have a relevant effect on cell proliferation and vitality and, therefore, can be recommended for the improvement of the properties of a biomaterial.
Assuntos
Celulose/química , Materiais Revestidos Biocompatíveis/química , Difosfonatos/farmacologia , Osteoblastos/efeitos dos fármacos , Alicerces Teciduais/química , Fosfatase Alcalina/metabolismo , Biomarcadores/metabolismo , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Celulose/ultraestrutura , Difosfonatos/metabolismo , Relação Dose-Resposta a Droga , Feto/citologia , Feto/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Humanos , Técnicas In Vitro , Osteoblastos/metabolismo , Osteoblastos/fisiologia , Osteoblastos/ultraestrutura , Osteocalcina/genética , Osteocalcina/metabolismo , Pamidronato , Silicatos/química , Fatores de TempoRESUMO
The aim of the study was to assess the suitability of different Ti-6Al-4V surfaces produced by the electron beam melting (EBM) process as matrices for attachment, proliferation, and differentiation of human fetal osteoblasts (hFOB 1.19). Human osteoblasts were cultured in vitro on smooth and rough-textured Ti-6Al-4V alloy disks. By means of cell number and vitality and SEM micrographs cell attachment and proliferation were observed. The differentiation rate was examined by using quantitative real-time PCR analysis for the gene expression of alkaline phosphatase (ALP), type I collagen (Coll-I), bone sialoprotein (BSP) and osteocalcin (OC). After 3 days of incubation there was a significant higher vitality (p < 0.02) and proliferation (p < 0.02) of hFOB cells on smooth surfaces (R(a) = 0.077 microm) and compact surfaces with adherent partly molten titanium particles on the surface (R(a) = 24.9 microm). On these samples cells spread over almost the whole surface. On porous surfaces with higher R(a) values, cell proliferation was reduced significantly. Quantitative real-time PCR analysis showed that the expression of osteogenic differentiation markers was not influenced by surface characteristics. Gene expression did not differ more than twofold for the different samples. Compact titanium samples with adherent partly molten titanium particles on the surface (R(a) = 24.9 microm) fabricated by the EBM process turned out to be best suited for cell proliferation, while highly rough surfaces (R(a) >/= 56.9 microm) reduced proliferation of hFOB cells. Surface characteristics of titanium can easily be changed by EBM in order to further improve proliferation.
Assuntos
Materiais Biocompatíveis/metabolismo , Osteoblastos/citologia , Osteoblastos/metabolismo , Titânio/química , Ligas , Anisotropia , Osso e Ossos/embriologia , Diferenciação Celular , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Elétrons , Regulação da Expressão Gênica , Humanos , Propriedades de Superfície , Temperatura , Titânio/farmacologiaRESUMO
The objective of this clinical trial was the analysis of 2 methods for engineering of autologous bone grafts for maxillary sinus augmentation with secondary implant placement. Group 1 (8 patients, 12 sinuses): cells of mandibular periosteum were cultured in a good manufacturing practice laboratory (2 weeks) with autologous serum and then transferred onto a collagen matrix. After another week, these composites were transplanted into the sinuses. In group 2A (2 patients, 3 sinuses), cells of maxillary bone were cultivated with autologous serum for 2 weeks, seeded onto natural bone mineral (NBM, diameter [Ø] = 8 mm) blocks, and cultivated for another 1.5 months. These composites were transplanted into the sinuses. Group 2B (control, 3 patients, 5 sinuses) received NBM blocks alone. In the course of implant placement 6 (group 1) and 8 (group 2) months later, core biopsy were taken. Clinical follow-up period was 1 to 2.5 years in group 1 and approximately 7 years in groups 2A and 2B. New vital bone was found in all cases at median densities of 38% (n = 12) in group 1, 32% in group 2A (n = 3), and 25% in group 2B (n = 5). Differences between group 1 and 2B as well as 2A and 2B were statistically significant ( p = 0.025). No adverse effects were seen. All methods described were capable of creating new bone tissue with sufficient stability for successful implant placement.