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Osteoid formation in pore regions of cell-HA complexes implanted in nude mice at 4 weeks post-implantation (H&E staining). All blocks were acid-treated. A and B) A cell-free HA block. C and D) A non-induced (control) cell-HA complex. E and F) An osteoinduced cell-HA complex. A, C, and E) Low magnification (bar = 250 mm). B, D, and F) Higher magnification (bar = 100 mm). Similar observations were obtained in two other samples.
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A superporous (85%) hydroxyapatite (HA) block was recently developed to improve osteoconductivity, but it was often not clinically successful when used to treat periodontal osseous defects. The primary purpose of this study is to develop a clinically applicable tissue-engineered bone substitute using this HA block and human alveolar periosteum-deri...
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... a negative control, cell-free, acid-treated HA blocks were maintained under the same conditions and sub- sequently implanted subcutaneously. The osteoid for- mation in pore regions of cell-HA complexes in nude mice is shown in Figure 5. Cells of the recipient tissue invaded and were distributed equally in almost all pore regions of the acid-treated blocks at 4 weeks post-implantation (Figs. ...
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... HA blocks were maintained under the same conditions and sub- sequently implanted subcutaneously. The osteoid for- mation in pore regions of cell-HA complexes in nude mice is shown in Figure 5. Cells of the recipient tissue invaded and were distributed equally in almost all pore regions of the acid-treated blocks at 4 weeks post-implantation (Figs. 5A and 5B). In non-treated HA blocks, a few cells of recipient tissue and vascular- ization were found only in the peripheral pore regions, whereas cell-free dead spaces were found in central pore regions (data not ...
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... human periosteal cells were cultured in acid- treated blocks without receiving osteogenic induc- tion for 10 days and implanted subcutaneously for 4 weeks, a greater number of cells, which were believed to be donor human periosteal cells, were found in all pore regions (Figs. 5C and 5D). However, the eosin- ophilic ECM was less observed in most pore regions, including deep pore regions. In contrast, when human periosteal cells were induced in acid-treated blocks with the osteogenic-inducing agent for the last 6 days prior to implantation, the ECM was well observed in all pore regions, and immature osteoids were found ...
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... in the osteoinduction medium (C). A non- induced (A and B) or osteoinduced cell-HA complex (C) was fixed and examined under light microscopy after staining with crystal violet (A) or observed with SEM (B and C). Bar = 250 mm. Data are representative of three independent tests. some pore regions, especially the peripheral regions of the HA blocks (Figs. 5E and ...
Citations
... Periosteum-derived cells (PDCs) extracted from the periosteum tissue, also express mesenchymal stem cell markers, and have the potential to undergo osteogenic, chondrogenic and adipogenic differentiation (De Bari et al 2006, Wang et al 2010, van Gastel et al 2012. Currently, many researchers use a variety of biological materials carrying PDCs to repair animal bone defects (Kawase et al 2010, Roberts et al 2014. Periosteum accelerates healing in bone injuries, as it provides the adequate progenitor cells to help regenerate bone tissue. ...
The aim of this study was to prepare chitosan–collagen (CS/COL) scaffolds that could release fibroblast growth factor-2 (FGF-2) and bone morphogenetic protein 2 (BMP-2), and to study the effect of this scaffold on bone repair. By improving the double emulsion/solvent evaporation technique, BMP-2 was encapsulated in poly(lactic acid)–poly(ethylene glycol)–poly(lactic acid) (PELA) microcapsules, to the surface of which FGF-2 was attached. The CS/COL scaffold carrying the microcapsules was prepared by freeze-drying. Periosteum derived cells (PDCs) were extracted and cultured on the scaffolds to study their proliferation and differentiation on the scaffolds. In addition, the effects of the scaffolds were investigated on rats with skull defects by micro-computed tomography and histology. We successfully prepared PELA microcapsules with external adherence to FGF-2 and encapsulated with BMP-2. The CS/COL scaffolds were porous and PDCs adhered, proliferated and underwent osteogenic differentiation on the scaffolds. The sequential release of FGF-2/BMP-2 had better osteogenic efficacy than other groups. Our results suggest that CS/COL scaffolds that bind FGF-2 and BMP-2 in combination with PDCs could be a promising new strategy for tissue engineering periosteum.
... More to the point, dental applications of apatites include direct pulp capping [306,[386][387][388][389][390], dentin hypersensitivity treatments [164-166], using in endodontics [391][392][393][394][395][396][397][398][399][400][401][402][403][404][405][406][407][408], orthodontics [409][410][411][412][413][414][415][416][417], oral and maxillofacial surgery [290,336,, orthognathic surgery [445][446][447][448][449], prosthodontics [450][451][452][453][454][455][456][457][458][459][460] and periodontics [461][462][463][464][465][466][467][468][469][470][471][472][473][474][475][476][477][478]. ...
Dental caries, also known as tooth decay or a cavity, remains a major public health problem in the most communities even though the prevalence of disease has decreased since the introduction of fluorides for dental care. In addition, there is dental erosion, which is a chemical wear of the dental hard tissues without the involvement of bacteria. Besides, there are other dental losses, which may be of a medical (decay or periodontal disease), age (population aging), traumatic (accident) or genetic (disorders) nature. All these cases clearly indicate that biomaterials to fill dental defects appear to be necessary to fulfill customers' needs regarding the properties and the processing of the products. Bioceramics and glass-ceramics are widely used for these purposes, as dental inlays, onlays, veneers, crowns or bridges. Calcium orthophosphates (CaPO4) belong to bioceramics but they have some specific advantage over other types of bioceramics due to a chemical similarity to the inorganic part of both human and mammalian bones and teeth. Therefore, CaPO4 (both alone and as constituents of various formulations) are used in dentistry as both dental fillers and implantable scaffolds. This review provides brief knowledge on CaPO4 and describes in details current state-of-the-art on their applications in dentistry and dentistry-related fields. Among the recognized dental specialties, CaPO4 are most frequently used in periodontics; however, the majority of the publications on CaPO4 in dentistry are devoted to unspecified "dental" fields.
... More to the point, dental applications of apatites include direct pulp capping [306,[386][387][388][389][390], dentin hypersensitivity treatments [164-166], using in endodontics [391][392][393][394][395][396][397][398][399][400][401][402][403][404][405][406][407][408], orthodontics [409][410][411][412][413][414][415][416][417], oral and maxillofacial surgery [290,336,, orthognathic surgery [445][446][447][448][449], prosthodontics [450][451][452][453][454][455][456][457][458][459][460] and periodontics [461][462][463][464][465][466][467][468][469][470][471][472][473][474][475][476][477][478]. ...
Dental caries, also known as tooth decay or a cavity, remains a major public health problem in the most communities even though the prevalence of disease has decreased since the introduction of fluorides for dental care. In addition, there is dental erosion, which is a chemical wear of the dental hard tissues without the involvement of bacteria. Besides, there are other dental losses, which may be of a medical (decay or periodontal disease), age (population aging), traumatic (accident) or genetic (disorders) nature. All these cases clearly indicate that biomaterials to fill dental defects appear to be necessary to fulfill customers’ needs regarding the properties and the processing of the products. Bioceramics and glass-ceramics are widely used for these purposes, as dental inlays, onlays, veneers, crowns or bridges. Calcium orthophosphates (CaPO ) belong to bioceramics but they have some specific advantage over other types of bioceramics due to a chemical similarity to the inorganic part of both human and mammalian bones and teeth. Therefore, CaPO (both alone and as constituents of various formulations) are used in dentistry as both dental fillers and implantable scaffolds. This review provides brief knowledge on CaPO and describes in details current state-of-the-art on their applications in dentistry and dentistry-related fields. Among the recognized dental specialties, CaPO are most frequently used in periodontics; however, the majority of the publications on CaPO in dentistry are devoted to unspecified “dental” fields.
... 8 Traditional HA bioceramics, which possess good osteoconductivity and bioactivity, have been reported to combine with human periosteum-derived cells, acting as an osteogenic bone substitute for periodontal regenerative therapy. 36 However, traditional HA bioceramics lack the osteoinductive ability to induce osteogenic differentiation of stem cells and stimulate periodontal tissue regeneration, including bone and cementum. 37 Therefore, there is a requirement for modification of HA bioceramics to enhance their biological activity. ...
The surface structure of bioceramic scaffolds is crucial for its bioactivity and osteoinductive ability, and in recent years, human periodontal ligament stem cells have been certified to possess high osteogenic and cementogenic differential ability. In the present study, hydroxyapatite (HA) bioceramics with micro-nano-hybrid surface (mnHA [the hybrid of nanorods and microrods]) were fabricated via hydrothermal reaction of the α-tricalcium phosphate granules as precursors in aqueous solution, and the effects of mnHA on the attachment, proliferation, osteogenic and cementogenic differentiations of human periodontal ligament stem cells as well as the related mechanisms were systematically investigated. The results showed that mnHA bioceramics could promote cell adhesion, proliferation, alkaline phosphatase (ALP) activity, and expression of osteogenic/cementogenic-related markers including runt-related transcription factor 2 (Runx2), ALP, osteocalcin (OCN), cementum attachment protein (CAP), and cementum protein (CEMP) as compared to the HA bioceramics with flat and dense surface. Moreover, mnHA bioceramics stimulated gene expression of low-density lipoprotein receptor-related protein 5 (LRP5) and β-catenin, which are the key genes of canonical Wnt signaling. Moreover, the stimulatory effect on ALP activity and osteogenic and cementogenic gene expression, including that of ALP, OCN, CAP, CEMP, and Runx2 of mnHA bioceramics could be repressed by canonical Wnt signaling inhibitor dickkopf1 (Dkk1). The results suggested that the HA bioceramics with mnHA could act as promising grafts for periodontal tissue regeneration.
... The cells were examined at 4 days (n = 5), 12 days (n = 5), and 20 days (n = 5) postwounding, which were appropriate intervals to evaluate the initial phase, the late phase of the proliferation stage, and the remodeling stage, respectively. As described in a previous report, 27 surgical procedures were performed under standard aseptic conditions. After anesthesia with 10% chloral hydrate, two circular fullthickness skin defects (diameter = 10 mm) were prepared in each rat, and 30 min later the wounds were treated or not with the above agents. ...
The mesenchymal stem cell (MSC) supernatant is well known as a rich source of autologous cytokines and universally used for tissue regeneration in current clinical medicine. However, the limitation of conditioned medium used in open-wound repair compels the need to find a more sophisticated way to take advantage of the trophic factors of MSCs. We have now fabricated a three-dimensional membrane from freeze-dried bone marrow stromal cell-conditioned medium (FBMSC-CM) using a simple freeze-dried protocol. Scanning electron microscopy images (SEM) showed the microstructure of the FBMSC-CM membrane (FBMSC-CMM) resembling a mesh containing growth factors. ELISA was used to test the paracrine factors retained in the FBMSC-CMM, and the results indicated that FBMSC-CMM withheld over 80% of the paracrine factors. Live/dead assays were adopted to test the toxicity of the FBMSC-CMM on cultured rat dermal fibroblasts (RDFs), and the results confirmed its biological safety with low toxicity. Moreover, the FBMSC-CMM could significantly accelerate wound healing and enhance the neovascularization as well as epithelization via strengthening the trophic factors in the wound bed as determined by immunohistochemical staining. Thus, the ability to maintain paracrine factors and enhance the effectiveness of these growth factors in the wound as well as the simple procedure and economical materials required for production qualifies the FBMSC-CMM to be a candidate biomaterial for open-wound regeneration.
... After reaching cell confluence, the periosteal sheets were enzymatically digested to disperse periosteal cells. 10 The cells were further expanded with growth medium and seeded onto the PRF membranes. After 20 h of incubation, cells attached to the PRF were fixed with 2.5% glutaraldehyde solution, dehydrated with a series of ethanol and t-butanol washes, and freeze-dried. ...
Platelet-rich fibrin (PRF) was developed as an advanced form of platelet-rich plasma to eliminate xenofactors, such as bovine thrombin, and it is mainly used as a source of growth factor for tissue regeneration. Furthermore, although a minor application, PRF in a compressed membrane-like form has also been used as a substitute for commercially available barrier membranes in guided-tissue regeneration (GTR) treatment. However, the PRF membrane is resorbed within 2 weeks or less at implantation sites; therefore, it can barely maintain sufficient space for bone regeneration. In this study, we developed and optimized a heat-compression technique and tested the feasibility of the resulting PRF membrane. Freshly prepared human PRF was first compressed with dry gauze and subsequently with a hot iron. Biodegradability was microscopically examined in vitro by treatment with plasmin at 37°C or in vivo by subcutaneous implantation in nude mice. Compared with the control gauze-compressed PRF, the heat-compressed PRF appeared plasmin-resistant and remained stable for longer than 10 days in vitro. Additionally, in animal implantation studies, the heat-compressed PRF was observed at least for 3 weeks postimplantation in vivo whereas the control PRF was completely resorbed within 2 weeks. Therefore, these findings suggest that the heat-compression technique reduces the rate of biodegradation of the PRF membrane without sacrificing its biocompatibility and that the heat-compressed PRF membrane easily could be prepared at chair-side and applied as a barrier membrane in the GTR treatment. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2014.
... The surfaces of the samples were coated with gold-palladium using a sputter coater (MSP-1S; Vacuum Device, Mito, Japan) and examined using a scanning electron microscopy (SEM, TM-1000; Hitachi, Tokyo, Japan) operating at an accelerating voltage of 15 kV. 21 ...
An atmospheric-pressure plasma (APP) treatment was recently reported to render titanium (Ti) surfaces more suitable for osteoblastic cell proliferation and osteogenesis. However, the mechanism of action remains to be clearly demonstrated. In this study, we focused on cell adhesion and examined the effects of the APP treatment on the initial responses of human prenatal-derived osteoblastic cells incubated on chemically polished commercially pure Ti (CP-cpTi) plates. In the medium containing 1% fetal bovine serum, the initial cell adhesion and the actin polymerization were evaluated by scanning electron microscopy and fluorescence microscopy. The expression of cell adhesion-related molecules and osteoblast markers at the messenger RNA level was assessed by real-time quantitative polymerase chain reaction. Although the cells on the APP-treated CP-cpTi surface developed fewer cytoskeletal actin fibers, they attached with higher affinity and consequently proliferated more actively (1.46-fold over control at 72 h). However, most of the cell adhesion molecule genes were significantly downregulated (from 40 to 85% of control) in the cells incubated on the APP-treated CP-cpTi surface at 24 h. Similarly, the osteoblast marker genes were significantly downregulated (from 49 to 63% of control) at 72 h. However, the osteoblast marker genes were drastically upregulated (from 197 to 296% of control) in these cells by dexamethasone and β-glycerophosphate treatment. These findings suggest that the APP treatment improves the ability of the CP-cpTi surface to support osteoblastic proliferation by enhancing the initial cell adhesion and supports osteoblastic differentiation when immature osteoblasts begin the differentiation process. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2014.
... After the periosteum-derived cell sheets achieved a diameter of 40e50 mm, the sheets were detached with a solution of 0.05% trypsin in 0.53 mmol/L ethylenediaminetetraacetic acid (EDTA) (Life Technologies) and dispersed for subculture (14). The dispersed periosteal cells were maintained in the growth medium until the cells reached sub-confluence. ...
Cultured human periosteal sheets more effectively function as an osteogenic grafting material at implantation sites than do dispersed periosteal cells. Because adherent cell growth and differentiation are regulated by cell-cell and cell-extracellular matrix contacts, we hypothesized that this advantage is a result of the unique cell adhesion pattern formed by their multiple cell layers and abundant extracellular matrix. To test this hypothesis, we prepared three distinct forms of periosteal cell cultures: three-dimensional cell-multilayered periosteal sheets, two-dimensional dispersed cell cultures, and three-dimensional hybrid mock-ups of cells dispersed onto collagen sponges.
Periosteal cells were obtained from human alveolar bone. Cell adhesion and extracellular matrix molecules were quantitatively determined at the messenger RNA and protein levels by means of real-time quantitative polymerase chain reaction and flow cytometry, respectively.
Real-time quantitative polymerase chain reaction analysis demonstrated that regardless of culture media α1 integrin, vascular cell adhesion molecule-1, fibronectin and collagen type 1 were substantially upregulated, whereas CD44 was strongly downregulated in periosteal sheets compared with dispersed cell monolayers. With increased thickness, stem cell medium upregulated several integrins (β1, α1 and α4), CD146, vascular cell adhesion molecule-1, fibronectin and collagen type 1 in the periosteal sheets. Flow cytometric analysis revealed that the active configuration of β1 integrin was substantially downregulated in the stem cell medium-expanded cell cultures. The cell adhesion pattern found in the mock-up cultures was almost identical to that of genuine periosteal sheets.
Integrin α1β1 and CD44 function as the main cell adhesion molecule in highly cell-multilayered periosteal sheets and dispersed cells, respectively. This difference may account for the more potent osteogenic activity shown by the thicker periosteal sheets.
... If cultured PDCs were used as a sheet, it might have the same mode of action as a guided bone regeneration (GBR) membrane or space filler, which prevents the intrusion of epithelial cells. 10,[30][31][32] Given that a recent study reported that the application of PDCs, together with the GBR technique, did not provide additional benefit to peri-implant bone regeneration, 33 the advantage of using cultured PDCs might be to exclude extra complications that could occur and thus favor new bone formation with the quintessential tissue. Additional study is necessary to verify the significance and contribution of having ' 'some cells' ' in cultured transplants of PDCs. ...
Clinically, bone marrow stromal cells (BMCs) are the most common source of osteoprogenitor cells. Its harvest process, however, is invasive to patients. Previous reports have shown the potential advantages of using periosteum-derived cells (PDCs) as a source of cell-based transplant therapy. The objective of our study was to characterize the osteoblastic differentiation and mineralization ability of PDCs versus BMCs and osteoblasts (OBs).
BMCs, OBs, and PDCs were isolated from 4-week-old male Wistar rats. To characterize the differentiation ability of the cells, MTS assay, alkaline phosphatase (ALP) activity staining, picrosirius red staining, and alizarin red staining were performed. Immunohistochemistry was performed on paraffin sections of calvarial periosteum to determine the presence of mesenchymal stem cells.
PDCs showed the greatest proliferation rate compared with BMCs and OBs. Matured collagenous matrix formation was observed in PDCs and BMCs. ALP-positive cells and in vitro mineralization were evident in all cell types analyzed; however, that of PDCs was not comparable to that of the OBs and BMCs. Immunohistochemistry revealed the presence of STRO-1-and CD105-positive cells in the cambium layer of the periosteum.
PDCs have remarkable proliferative ability, but contain only a small population of osteogenic cells compared with BMCs and OBs. Although cell activity can be affected by various factors, such as age, culture condition, additives, and so forth, PDCs are likely not the source of OBs, although they might provide matrices that indirectly aid in bone formation.
... [3][4][5] MSC from various tissues have been investigated for cell-based regeneration of periodontal tissues, including the periodontal ligament, 6-8 bone marrow, [9][10][11] adipose tissue, 12 and periosteum. [13][14][15] Cultured stem cells from the periodontal ligament (PDLSCs) possess characteristics of MSC such as cell surface MSC marker expression (CD105 + , CD90 + , CD44 + , CD73 + , CD45 -, CD31 -, and CD34 -) and multilineage differentiation ability (i.e., into osteoblasts, chondrocytes, and adipocytes). [16][17][18] In addition to MSC characteristics, PDLSCs have unique properties to form a cementum/ periodontal ligament complex-like structure when ectopically transplanted in animals. ...
Periodontal disease is characterized by the destruction of tooth supporting tissues. Regeneration of periodontal tissues using ex vivo expanded cells has been introduced and studied, although appropriate methodology has not yet been established. We developed a novel cell transplant method for periodontal regeneration using periodontal ligament stem cell (PDLSC)-transferred amniotic membrane (PDLSC-amnion). The aim of this study was to investigate the regenerative potential of PDLSC-amnion in a rat periodontal defect model. Cultured PDLSCs were transferred onto amniotic membranes using a glass substrate treated with polyethylene glycol and photolithography. The properties of PDLSCs were investigated by flow cytometry and in vitro differentiation. PDLSC-amnion was transplanted into surgically created periodontal defects in rat maxillary molars. Periodontal regeneration was evaluated by micro-CT and histological analysis. PDLSCs showed mesenchymal stem cell-like characteristics such as cell surface marker expression (CD90, CD44, CD73, CD105, CD146 and STRO-1) and tri-lineage differentiation ability (i.e., into osteoblasts, adipocytes and chondrocytes). PDLSC-amnion exhibited a single layer of PDLSCs on the amniotic membrane and stability of the sheet even with movement and deformation caused by surgical instruments. We observed that the PDLSC-amnion enhanced periodontal tissue regeneration as determined by micro-CT and histology by 4 weeks after transplantation. These data suggest that PDLSC-amnion has therapeutic potential as a novel cell based regenerative periodontal therapy.
