Figure 6.
Figure 6. Cultivation and osteogenic differentiation of DFCs on PA after modification with collagen I. (Left) Relative cell number and (Right) normalized ALP activity.
Figure 5. resentative results are shown for dNC-PCs.
Figure 5. Evaluation of osteogenic differentiation. (A) Clustergram of PCR-array results; (B-C) histology of differentiated dental cells on AP (B) and SB (C). Representative results are shown for dNC-PCs.
Figure 4. dishes for control.
Figure 4. Osteogenic differentiation of dental stem cells. Normalized ALP activity of dNC-PCs and DFCs on AP and SB (A) and on silicone (B). Cells were differentiated on standard cell culture dishes for control.
Figure 3. (black number), apoptotic cells (blue number), and dead cells (red number). (B) Western blot analyses show the expression of the pro-apoptotic marker BAX and the anti-apoptotic marker BCL2.
Figure 3. Evaluation of programmed cell death (apoptosis) in dental stem cells. (A) Flow cytometry analyses (for details materials and methods) show percentage of vital cells (black number), apop...
Figure 2. spheroid cell clusters on silicone (representative pictures for DFCs); Silicone (24 and 48 h).
Figure 2. C ell proliferation of dNC-PCs and DFCs on tested materials. (A) and (B) Relative cell numbers; (C) spheroid cell clusters on silicone (representative pictures for DFCs); Silicone (24 and 48 h).
Figure 1. ative pictures of DFCs.
Figure 1. Cell attachment on tested materials. (A) Relative cell adherence of DFCs and dNC-PCs; (B) dental cells did little adhere on PA; representative pictures of DFCs.
Gosau, M., Viale-Bouroncle, S., Eickhoff, H. et al. Evaluation of implant-materials as cell carriers for dental stem cells under in vitro conditions.
Int J Implant Dent 1, 2 (2015). https://doi.org/10.1186/s40729-014-0002-y
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Received: 17 September 2014
Accepted: 20 November 2014
Published: 12 February 2015
DOI: https://doi.org/10.1186/s40729-014-0002-y
DFCs and dNC-PCs expressed typical markers for dental stem cells.
Martin Gosau, Sandra Viale-Bouroncle, Hannah Eickhoff, Esthera Prateeptongkum, Anja Reck, W Götz, Christoph Klingelhöffer, Steffen Müller and Christian Morsczeck declare that they have no competing interests.
SVB, HE, and EP carried out all cell biology experiments, performed real-time PCRs, Western blots, and the statistical analysis and made figures for the manuscript. AR carried out and ana...
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Christian Morsczeck.
Department of Cranio- and Maxillofacial Surgery, Hospital of the University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053, Regensburg, Germany
Martin Gosau, Sandra Viale-Bouroncle, Hannah Eickhoff, Esthera Prateeptongkum, Anja Reck, Christoph Klingelhöffer, Steffen Müller & Christian Morsczeck
Department of Oral and Maxillofacial Surgery, Paracelsus Medical University Nuernberg, B...
This study was supported by a grant from the Deutschen Gesellschaft für Implantologie (DGI) e.V.
Marino G, Rosso F, Cafiero G, Tortora C, Moraci M, Barbarisi M, Barbarisi A. Beta-tricalcium phosphate 3D scaffold promote alone osteogenic differentiation of human adipose stem cells: in vitro study. J Mater Sci Mater Med. 2010; 21:353–63.
Arnold U, Lindenhayn K, Perka C. In vitro-cultivation of human periosteum derived cells in bioresorbable polymer-TCP-composites. Biomaterials. 2002; 23:2303...
Felthaus O, Gosau M, Morsczeck C. ZBTB16 induces osteogenic differentiation marker genes in dental follicle cells independent from RUNX2.
J Periodontol. 2014; 85:e144–51.
Viale-Bouroncle S, Gosau M, Küpper K, Möhl C, Brockhoff G, Reichert TE, Schmalz G, Ettl T, Morsczeck C. Rigid matrix supports osteogenic differentiation of stem cells from human exfoliated deciduous teeth (SHED).
Differ Re...
Kagami H, Agata H, Tojo A. Bone marrow stromal cells (bone marrow-derived multipotent mesenchymal stromal cells) for bone tissue engineering: basic science to clinical translation.
Int J Biochem Cell Biol. 2011; 43:286–89.
Robey PG. Cell sources for bone regeneration: the good, the bad, and the ugly (but promising).
Tissue Eng Part B Rev. 2011; 17:423–30.
Morsczeck C, Shi S, Huang G. Stem/...
In conclusion, our work supports our hypothesis that soft implant materials are not suitable for dental tissue engineering. Moreover, our study also supports the results of our previous studies with DFCs and TCP that induction of apoptosis did not impair the proliferation and the differentiation in dental stem cells.
In a previous study, we showed that TCP induced the programmed cell death (apoptosis) in DFCs [11]. Our new study investigated therefore the induction of apoptosis in dental cells. While SB and soft materials did not induce apoptosis or cell death, AP induced obviously cell death and apoptosis in dental cells. Here, the results for dNC-PCs and DFCs were almost the same. Interestingly, neither sili...
Scaffolds play an important role in tissue engineering. However, little is known about the proliferation and differentiation of DFCs and dNC-PCs on different types of materials. As we have learned from previous studies mechanical properties such as surface stiffness are decisive for a successful osteogenic differentiation of dental stem cells [13,14]. Moreover, we showed that bone substitute mater...
The soft material PA was also treated with the extracellular matrix protein collagen to improve cell adherence. We tested representatively DFCs with collagen I modified PA. DFCs adhered and proliferated on modified PA, but, however, the specific ALP activity was reduced in comparison to that of DFCs on standard cell culture dishes (Figure 6). This reduction of the specific ALP activity was simila...
Dental cells were cultivated in standard cell culture media until passage 6. Cell adherence and cell proliferation/growth were measured for the estimation of cell viability on tested rigid and soft materials. In Figure 1, the cell adherence of dNC-PCs on bone substitute materials was better than that of DFCs. However, both dental cells types adhered very well on silicone. Unluckily, dental cells ...
DFCs were cultivated until sub-confluence (>80%) in standard cell culture medium before the differentiation starts with the osteogenic differentiation medium (ODM) comprised DMEM (PAA) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 100 μmol/L ascorbic acid 2-phosphate, 10 mmol/L KH2PO4, 1 × 10−8 mol/L dexamethasone sodium phosphate (Sigma-Aldrich, St. Louis, MO, USA), HEPES (20 ...
Numbers of vital cells were evaluated after days 1, 2, 3, and 6. For cell counting, cell cultures were incubated with the cell counting kit 8 (CCK8) ready to use solution according to manufactures instructions (Dojindo, Rockville, MD, USA). The optical density (O.D.) was measured at a wavelength of 450 nm. For the evaluation of the cell adherence (normalized to standard cell culture dishes), cell...
The bone substitutes Maxgraft® (AP) and Maxresorb® (SB) were obtained from the company Botiss (botiss dental GmbH, Berlin, Germany). Maxgraft® is a sterile, high-safety allograft product (AP), derived from human donor bone. It is processed by an audited and certified bone bank (Cells+ Tissue Bank Austria, Berlin, Germany). In contrast, Maxresorb® is a fully synthetic bone graft substitute (SB)...
The isolation and characterization of DFCs and dNC-PCs were described in previous studies [4,7,12]. DFCs were routinely cultivated in DMEM (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and 100 μg/ml penicillin/streptomycin (standard cell culture medium). dNC-PCs were cultivated in DMEM (Sigma-Aldrich) supplemented with 15% fetal ...
Unfortunately, an additional study showed that TCP induced apoptosis in DFCs [11]. However, the induction of apoptosis exposed a risk for cellular therapies. We decided therefore to evaluate additional implant materials for the identification of a suitable scaffold for dental stem cells. Soft materials such as silicone are successfully used in regenerative medicine, and they are suitable for tissu...
While bone substitute materials are routinely used, especially vertical bone, augmentation of the jaws is still a problematic step. Dental stem cells in combination with bone substitute materials may accelerate the augmentation of alveolar bone and perhaps, stem cell-based therapies can become an alternative to autologous, allogenic, or synthetic bone transplants and substitutes [1,2]. However, sc...
Dental stem cells in combination with implant materials may become an alternative to autologous bone transplants. For tissue engineering different types of soft and rigid implant materials are available, but little is known about the viability and the osteogenic differentiation of dental stem cells on these different types of materials. According to previous studies we proposed that rigid bone sub...
Fig. 4. e. Margins between bone defect and scaffold are less demarcated due to new bone growth
Fig. 4. a Peri-apical X-ray of zirconia scaffold immediately placed in bone defect. Margins between scaffold and bone are clearly demarcated. b Peri-apical X-ray of zirconia scaffold after completion of healing time. Margins between bone defect and scaffold are less demarcated due to new bone growth
Fig. 3. hite arrow). Mineralized bone formation (black arrow) was less dense compared to HA-enriched scaffolds. b Histological section showing different sizes of pores present in porous zirconia scaffolds (Control specimen). Mineralization started by lining pore walls (white arrow). Unmineralized bone stained blue
Fig. 3. a Histological section demonstrating bone growth in control zirconia sc...
Fig. 2. zirconia scaffold (black arrow). Unmineralized bone stained blue. Almost entire surface porosity was filled with new dense bone. b Histological section demonstrating bone growth in HA-enriched zirconia scaffold starting from the periphery of the surgical wound (white arrow). Islands of entrapped HA particles were surrounded by mineralized boney matrix (black arrow) which were identified ...
Fig. 1. rating agglomeration of nano-hydroxyapatite particles filling the porous structure
Fig. 1. a SEM image, ×10,000, demonstrating internal porosity of the fabricated zirconia scaffolds. b SEM image, ×30,500, demonstrating agglomeration of nano-hydroxyapatite particles filling the porous structure
Aboushelib, M.N., Shawky, R. Osteogenesis ability of CAD/CAM porous zirconia scaffolds enriched with nano-hydroxyapatite particles. Int J Implant Dent 3, 21 (2017). https://doi.org/10.1186/s40729-017-0082-6
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Received: 15 November 2016
Accepted: 08 May 2017
Published: 19 May 2017
DOI: https://doi.org/10.1186/s40729-017-0082-6
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were m...
Dental Biomaterials Department, Faculty of Dentistry, Alexandria University, Champollion st, Azarita, Alexandria, Egypt
Moustafa N. Aboushelib
Oral Surgery Department, Faculty of Dentistry, Alexandria University, Alexandria, Egypt
Rehab Shawky
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You can also search for this author in PubMed Google Scholar
Correspondence to Moustafa...
Part of this study was supported by STDF reintegration grant number 489 and was performed in collaboration with the Oral Surgery Department, Faculty of Dentistry, Alexandria University, Egypt.
MNA prepared the specimens and performed the characterization process. He participated in preparation of histological section, data collection and analysis, and in preparation of the manuscript. RS performe...
Naleway SE, Fickas KC, Maker YN, Meyers MA, McKittrick J. Reproducibility of ZrO2-based freeze casting for biomaterials. Mater Sci Eng C Mater Biol Appl. 2016;61:105–12.
Song YG, Cho IH. Characteristics and osteogenic effect of zirconia porous scaffold coated with beta-TCP/HA. J Adv Prosthodont. 2014;6:285–94.
Okano T, et al. Absorbed and effective doses from cone beam volumetric imaging for...
Chiapasco M, Casentini P, Zaniboni M. Bone augmentation procedures in implant dentistry. Int J Oral Maxillofac Implants. 2009;24:237–59.
Rocchietta I, Fontana F, Simion M. Clinical outcomes of vertical bone augmentation to enable dental implant placement: a systematic review. J Clin Periodontol. 2008;35:203–15.
Louis PJ, Gutta R, Said-Al-Naief N, Bartolucci AA. Reconstruction of the maxilla ...
Within the limitations of this study, hydroxyapatite enhanced osteogenesis ability of porous zirconia scaffolds.
Histomorphometric analysis revealed that bone growth start to develop as early as 6 weeks by lining pore cavity walls. Mineralized bone matrix was observed to penetrate 1–2 mm under the surface of the scaffolds thus providing mechanical stability of the inserted prosthesis. Healing continued by filling the entire pore volume (Fig. 2a, b). The presence of nano-hydroxyapatite particles enhanced...
Porous scaffolds are designed to allow ingrowth of the surrounding bone within the internal porosity of the solid matrix. Different types of bioactive materials were mixed with zirconia to enhance bone formation. Two sizes of pores were incorporated in the structure of the fabricated scaffolds. Micro-pores in range of 50 μm constituted the majority of the entire pore volume (50 wt.%) of the fabr...
Mercury porosimetery revealed comparable (F = 0.057, P
Six weeks after insertion of the scaffolds, the animals were given an over dose of an anesthetic injection and section blocks were obtained by cutting the mandible maintaining 10 mm of sound bone around the scaffolds. Cut sections were immediately fixed in 4% buffered formaldehyde and dehydrated in graded ethanol solutions using a dehydration system under agitation and vacuum, and the specimens w...
Nano-hydroxyapatite particles were prepared using sol gel chemical precipitation method. The sol was thermally aged at low temperature at 50 °C for 2 h. Upon drying the sol particles agglomerated into a dry gel through van der Waals forces composed of 10–14-nm particles. A crystalline apatite is achieved after sintering at 450 °C resulting in a gained structure of 25–55 nm in diameter. T...
Zirconia powder (50 μm, 3 mol YTZP, E grade, Tosoh, Japan) was mixed with 50 wt.% resin beads (50 μm polymethyl methacrylate powder) added to create microscopic pore sizes. Thirty weight percent coarse sodium chloride particles (500–700 μm) were added to the mix to create large interconnected pores. The powder was mixed in a rotating cylinder for 24 h to insure homogenous powder distribu...
Modern radiographic imaging techniques in combination with advanced computer designing software could reconstruct a three-dimensional image of large boney defects [16]. Designing the shape of the required scaffold could easily be performed to accurately fit the available defect size using advanced imaging and designing software [17]. Finally, the required shape could be directly milled from differ...
Principles of tissue engineering are used today in an attempt to reconstruct damaged human tissue. In the dental field, several types of bone grafting materials are currently available which could be directly used to augment atrophic jaws before implant placement. However, the main drawback of these grafts is related to difficulty of preserving the required shape of the graft during the healing ti...
The aim of this study was to evaluate osteogenesis ability of CAD/CAM porous zirconia scaffolds enriched with hydroxy apatite used to augment large boney defects in a dog model.
Surgical defects were made bilaterally on the lower jaw of 12 Beagle dogs. Cone beam CT images were used to create three dimensional images of the healed defects. Porous zirconia scaffolds were fabricated by milling custo...
Aboushelib, M.N., Shawky, R. Osteogenesis ability of CAD/CAM porous zirconia scaffolds enriched with nano-hydroxyapatite particles.
Int J Implant Dent 3, 21 (2017). https://doi.org/10.1186/s40729-017-0082-6
Download citation
Received: 15 November 2016
Accepted: 08 May 2017
Published: 19 May 2017
DOI: https://doi.org/10.1186/s40729-017-0082-6
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
Dental Biomaterials Department, Faculty of Dentistry, Alexandria University, Champollion st, Azarita, Alexandria, Egypt
Moustafa N. Aboushelib
Oral Surgery Department, Faculty of Dentistry, Alexandria University, Alexandria, Egypt
Rehab Shawky
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...
Part of this study was supported by STDF reintegration grant number 489 and was performed in collaboration with the Oral Surgery Department, Faculty of Dentistry, Alexandria University, Egypt.
MNA prepared the specimens and performed the characterization process. He participated in preparation of histological section, data collection and analysis, and in preparation of the manuscript. RS performe...
Naleway SE, Fickas KC, Maker YN, Meyers MA, McKittrick J. Reproducibility of ZrO2-based freeze casting for biomaterials. Mater Sci Eng C Mater Biol Appl. 2016;61:105–12.
Song YG, Cho IH. Characteristics and osteogenic effect of zirconia porous scaffold coated with beta-TCP/HA. J Adv Prosthodont. 2014;6:285–94.
Okano T, et al. Absorbed and effective doses from cone beam volumetric imaging for...
Chiapasco M, Casentini P, Zaniboni M. Bone augmentation procedures in implant dentistry. Int J Oral Maxillofac Implants. 2009;24:237–59.
Rocchietta I, Fontana F, Simion M. Clinical outcomes of vertical bone augmentation to enable dental implant placement: a systematic review. J Clin Periodontol. 2008;35:203–15.
Louis PJ, Gutta R, Said-Al-Naief N, Bartolucci AA. Reconstruction of the maxilla ...
Within the limitations of this study, hydroxyapatite enhanced osteogenesis ability of porous zirconia scaffolds.
Histomorphometric analysis revealed that bone growth start to develop as early as 6 weeks by lining pore cavity walls. Mineralized bone matrix was observed to penetrate 1–2 mm under the surface of the scaffolds thus providing mechanical stability of the inserted prosthesis. Healing continued by filling the entire pore volume (Fig. 2a, b). The presence of nano-hydroxyapatite particles enhanced...
Porous scaffolds are designed to allow ingrowth of the surrounding bone within the internal porosity of the solid matrix. Different types of bioactive materials were mixed with zirconia to enhance bone formation. Two sizes of pores were incorporated in the structure of the fabricated scaffolds. Micro-pores in range of 50 μm constituted the majority of the entire pore volume (50 wt.%) of the fabr...
Mercury porosimetery revealed comparable (F = 0.057, P
Six weeks after insertion of the scaffolds, the animals were given an over dose of an anesthetic injection and section blocks were obtained by cutting the mandible maintaining 10 mm of sound bone around the scaffolds. Cut sections were immediately fixed in 4% buffered formaldehyde and dehydrated in graded ethanol solutions using a dehydration system under agitation and vacuum, and the specimens w...
Nano-hydroxyapatite particles were prepared using sol gel chemical precipitation method. The sol was thermally aged at low temperature at 50 °C for 2 h. Upon drying the sol particles agglomerated into a dry gel through van der Waals forces composed of 10–14-nm particles. A crystalline apatite is achieved after sintering at 450 °C resulting in a gained structure of 25–55 nm in diameter. T...
Zirconia powder (50 μm, 3 mol YTZP, E grade, Tosoh, Japan) was mixed with 50 wt.% resin beads (50 μm polymethyl methacrylate powder) added to create microscopic pore sizes. Thirty weight percent coarse sodium chloride particles (500–700 μm) were added to the mix to create large interconnected pores. The powder was mixed in a rotating cylinder for 24 h to insure homogenous powder distribu...
Modern radiographic imaging techniques in combination with advanced computer designing software could reconstruct a three-dimensional image of large boney defects [16]. Designing the shape of the required scaffold could easily be performed to accurately fit the available defect size using advanced imaging and designing software [17]. Finally, the required shape could be directly milled from differ...
Principles of tissue engineering are used today in an attempt to reconstruct damaged human tissue. In the dental field, several types of bone grafting materials are currently available which could be directly used to augment atrophic jaws before implant placement. However, the main drawback of these grafts is related to difficulty of preserving the required shape of the graft during the healing ti...
The aim of this study was to evaluate osteogenesis ability of CAD/CAM porous zirconia scaffolds enriched with hydroxy apatite used to augment large boney defects in a dog model.
Surgical defects were made bilaterally on the lower jaw of 12 Beagle dogs. Cone beam CT images were used to create three dimensional images of the healed defects. Porous zirconia scaffolds were fabricated by milling custo...
Fig. 11. Energy dispersive spectrum of control Cp titanium specimen
Fig. 11. Energy dispersive spectrum of control Cp titanium specimen
Fig. 10. Energy dispersive spectrum of Cp titanium specimen coated with HA-Zn
Fig. 10. Energy dispersive spectrum of Cp titanium specimen coated with HA-Zn
Fig. 9. Scanning electron microphotograph of control Cp titanium specimen at ×20,000
Fig. 9. Scanning electron microphotograph of control Cp titanium specimen at ×20,000
Fig. 8. Scanning electron microphotograph of control Cp titanium specimen at ×10,000
Fig. 8. Scanning electron microphotograph of control Cp titanium specimen at ×10,000
Fig. 7. Scanning electron microphotograph of control Cp Titanium specimen at X 5,000
Fig. 7. Scanning electron microphotograph of control Cp Titanium specimen at X 5,000
Fig. 6. Scanning electron microphotograph of Cp titanium specimen coated with HA-Zn at ×20,000
Fig. 6. Scanning electron microphotograph of Cp titanium specimen coated with HA-Zn at ×20,000
Fig. 5. Scanning electron microphotograph of Cp Titanium specimen coated with HA-Zn at X10,000
Fig. 5. Scanning electron microphotograph of Cp Titanium specimen coated with HA-Zn at X10,000
Fig. 4. Scanning electron microphotograph of Cp titanium specimen coated with nano HA- Zn at ×5000
Fig. 4. Scanning electron microphotograph of Cp titanium specimen coated with nano HA- Zn at ×5000
Fig. 3. IR spectra of HA-Zn powder scrapped from coated titanium specimen
Fig. 3. IR spectra of HA-Zn powder scrapped from coated titanium specimen
Fig. 2. IR spectra of Ca(NO3)2·4 H2O powder prepared from a natural source (CB)
Fig. 2. IR spectra of Ca(NO3)2·4 H2O powder prepared from a natural source (CB)
Fig. 1. Graphical presentation of the electrochemical-deposition coating process’ equipment
Fig. 1. Graphical presentation of the electrochemical-deposition coating process’ equipment
Number of specimens
Mean ± (SD)
Standard error mean
F value
P value
Control
7...
El-Wassefy, N.A., Reicha, F.M. & Aref, N.S. Electro-chemical deposition of nano hydroxyapatite-zinc coating on titanium metal substrate.
Int J Implant Dent 3, 39 (2017). https://doi.org/10.1186/s40729-017-0095-1
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Received: 20 March 2017
Accepted: 28 July 2017
Published: 13 August 2017
DOI: https://doi.org/10.1186/s40729-017-0095-1
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
El-Wassefy N, Aref N, and Reicha F declare that they have no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Dental Biomaterials Department, Faculty of Dentistry, Mansoura University, 35516 El Gomhoria St., Mansoura, Egypt
N. A. El-Wassefy & N. S. Aref
Physics Department, Faculty of science, Mansoura University, 35516 El Gomhoria St., Mansoura, Egypt
F. M. Reicha
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The authors would like to express their gratitude for Dr. Sherif Kishk, Professor of Communication and Electrical Engineering, Faculty of Engineering, Mansoura University, for his help in photographing and analyzing the coating for adhesion test.
Kuo MC, Yen SK. The process of electrochemical deposited hydroxyapatite coatings on biomedical titanium at room temperature. Mater Sci Eng C. 2002;20:153–60.
Suchanek W, Yoshimura M. Processing and properties of hydroxyapatite-based biomaterials for use as hard tissue replacement implants. J Mater Res. 1998;13:94–117.
Kohli S, Batra U, Kapoor S. Influence of zinc substitution on physicochemi...
Hosea HJ, Taylor CG, Wood T, Mollard R, Weiler HA. Zinc-deficient rats have more limited bone recovery during repletion than diet-restricted rats. Exp Biol Med. 2004;299:303–11.
Tsai M-T, Chang Y-Y, Huang H-L, Hsu J-T, Chen Y-C, Wu AY-J. Characterization and antibacterial performance of bioactive Ti–Zn–O coatings deposited on titanium implants. Thin Solid Films. 2013;528:143–50.
Hu H, Zh...
Brunette DM, Tengvall P, Textor M TP, Textor M, Thomsen P. Titanium in medicine: material science, surface science, engineering, biological responses and medical applications. Springer Science & Business Media; 2012. p.13–24.
Heydenrijk K, Meijer HJA, van der Reijden WA, Vissink A, Raghoebar GM, Stegenga B. Microbiota around root-formed endosseous implants. A review of the literature. October. ...
The electro-chemical method can be employed for HA-Zn coating deposition on titanium metal, where Ca source was a recycled cuttlebone fish to precipitate HA phases. Using a Zn anode on a low-sustained voltage was able to induce an even coat thickness of HA-Zn precipitation and increase the surface roughness significantly.
Yang et al. prepared a Zn-HA coating on Ti plates by an electrochemical process, and the SEM examination showed irregularly shaped rod-like crystals with hexagonal cross-section; this corresponded well with the current study results. They also concluded that a Zn-HA coating improves proliferation and differentiation of osteoblasts and would enhance implant osseointegration [11].
Ceramic coatings ...
Metallic orthopedic prosthesis is most commonly used due to its good mechanical properties, but its failure mostly occurs due to the lack of proper bone bonding and/or the occurrence of post-operative infections. Hydroxyapatite is commonly used as a bone filler biomaterial or as a coat for titanium prosthesis due to its decent biocompatibility, osseoconductivity, and bioactivity [26]. However, as ...
Following the examination of X cut areas after the adhesive tape removal; the adhesion was rated to be 5A, as no peeling or coat removal occurred along the incisions' length or at their intersection.
Figure 2 shows the FT-IR spectra of Ca(NO3)2·4H2O with weak sharp absorption peak bands at 742, 821, and 1048 cm−1, a strong broad absorption band at 1354 cm-1, and a strong shoulder absorption band at 1455 cm−1. A wide broad absorption band peak appears at 3442 cm−1 due to the presence of water. Figure 3 shows the FT-IR spectra of HA-Zn powder scrapped from CpTi specimens; the band a...
The coating was scrapped from Ti specimen's surface and investigated for its chemical structure using FT-IR spectroscopy. The powder was investigated by double-beam dispersive IR spectrometer (Nicolet iS10, Thermo Electron Corporation, UK) which utilized the selected range of 400 to 4000 wave numbers (cm−1) at 4 cm−1 resolution and averaging of 100 scans. Two milligrams of scrapped powder was...
Commercially pure Ti (CpTi) grade II specimens were cut down into plates with dimensions 10 × 10 × 2 mm and used as substrates (cathode material) for depositing HA and Zn. CpTi specimens were polished with successive grades of silicon carbide papers, ultra-sonicated in acetone (99.5%, EM Science), rinsed in distilled water, and then air dried at room temperature, before they were used f...
The aim of the present work was to develop well-adhered and uniform hydroxyapatite-zinc coatings on titanium metal substrate, through an in vitro electro-chemical deposition method. The coating was characterized for functional chemical group, surface morphology, surface chemical analysis, surface roughness, and coat adhesive bonding by Fourier transform infrared spectrometer (FT-IR), scanning elec...
Titanium metal is one of the most widely used biomedical orthopedic materials because of its decent mechanical properties [1]. However, as an inert material, it cannot induce osteogenesis and has no antibacterial properties [2]. In order to improve surface bioactivity of titanium substrates, numerous methods have been proposed to cover it with bio-ceramic coatings [1]. Various clinical studies ...
Titanium is an inert metal that does not induce osteogenesis and has no antibacterial properties; it is proposed that hydroxyapatite coating can enhance its bioactivity, while zinc can contribute to antibacterial properties and improve osseointegration.
A nano-sized hydroxyapatite-zinc coating was deposited on commercially pure titanium using an electro-chemical process, in order to increase its ...
Aboushelib, M.N., Shawky, R. Osteogenesis ability of CAD/CAM porous zirconia scaffolds enriched with nano-hydroxyapatite particles.
Int J Implant Dent 3, 21 (2017). https://doi.org/10.1186/s40729-017-0082-6
Download citation
Received: 15 November 2016
Accepted: 08 May 2017
Published: 19 May 2017
DOI: https://doi.org/10.1186/s40729-017-0082-6
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
Dental Biomaterials Department, Faculty of Dentistry, Alexandria University, Champollion st, Azarita, Alexandria, Egypt
Moustafa N. Aboushelib
Oral Surgery Department, Faculty of Dentistry, Alexandria University, Alexandria, Egypt
Rehab Shawky
You can also search for this author in
PubMed Google Scholar
You can also search for this author in
...
Part of this study was supported by STDF reintegration grant number 489 and was performed in collaboration with the Oral Surgery Department, Faculty of Dentistry, Alexandria University, Egypt.
MNA prepared the specimens and performed the characterization process. He participated in preparation of histological section, data collection and analysis, and in preparation of the manuscript. RS performe...
Naleway SE, Fickas KC, Maker YN, Meyers MA, McKittrick J. Reproducibility of ZrO2-based freeze casting for biomaterials. Mater Sci Eng C Mater Biol Appl. 2016;61:105–12.
Song YG, Cho IH. Characteristics and osteogenic effect of zirconia porous scaffold coated with beta-TCP/HA. J Adv Prosthodont. 2014;6:285–94.
Okano T, et al. Absorbed and effective doses from cone beam volumetric imaging for...
Chiapasco M, Casentini P, Zaniboni M. Bone augmentation procedures in implant dentistry. Int J Oral Maxillofac Implants. 2009;24:237–59.
Rocchietta I, Fontana F, Simion M. Clinical outcomes of vertical bone augmentation to enable dental implant placement: a systematic review. J Clin Periodontol. 2008;35:203–15.
Louis PJ, Gutta R, Said-Al-Naief N, Bartolucci AA. Reconstruction of the maxilla ...
Within the limitations of this study, hydroxyapatite enhanced osteogenesis ability of porous zirconia scaffolds.
Histomorphometric analysis revealed that bone growth start to develop as early as 6 weeks by lining pore cavity walls. Mineralized bone matrix was observed to penetrate 1–2 mm under the surface of the scaffolds thus providing mechanical stability of the inserted prosthesis. Healing continued by filling the entire pore volume (Fig. 2a, b). The presence of nano-hydroxyapatite particles enhanced...
Porous scaffolds are designed to allow ingrowth of the surrounding bone within the internal porosity of the solid matrix. Different types of bioactive materials were mixed with zirconia to enhance bone formation. Two sizes of pores were incorporated in the structure of the fabricated scaffolds. Micro-pores in range of 50 μm constituted the majority of the entire pore volume (50 wt.%) of the fabr...
Mercury porosimetery revealed comparable (F = 0.057, P
Six weeks after insertion of the scaffolds, the animals were given an over dose of an anesthetic injection and section blocks were obtained by cutting the mandible maintaining 10 mm of sound bone around the scaffolds. Cut sections were immediately fixed in 4% buffered formaldehyde and dehydrated in graded ethanol solutions using a dehydration system under agitation and vacuum, and the specimens w...
Nano-hydroxyapatite particles were prepared using sol gel chemical precipitation method. The sol was thermally aged at low temperature at 50 °C for 2 h. Upon drying the sol particles agglomerated into a dry gel through van der Waals forces composed of 10–14-nm particles. A crystalline apatite is achieved after sintering at 450 °C resulting in a gained structure of 25–55 nm in diameter. T...
Zirconia powder (50 μm, 3 mol YTZP, E grade, Tosoh, Japan) was mixed with 50 wt.% resin beads (50 μm polymethyl methacrylate powder) added to create microscopic pore sizes. Thirty weight percent coarse sodium chloride particles (500–700 μm) were added to the mix to create large interconnected pores. The powder was mixed in a rotating cylinder for 24 h to insure homogenous powder distribu...
Modern radiographic imaging techniques in combination with advanced computer designing software could reconstruct a three-dimensional image of large boney defects [16]. Designing the shape of the required scaffold could easily be performed to accurately fit the available defect size using advanced imaging and designing software [17]. Finally, the required shape could be directly milled from differ...
Principles of tissue engineering are used today in an attempt to reconstruct damaged human tissue. In the dental field, several types of bone grafting materials are currently available which could be directly used to augment atrophic jaws before implant placement. However, the main drawback of these grafts is related to difficulty of preserving the required shape of the graft during the healing ti...
The aim of this study was to evaluate osteogenesis ability of CAD/CAM porous zirconia scaffolds enriched with hydroxy apatite used to augment large boney defects in a dog model.
Surgical defects were made bilaterally on the lower jaw of 12 Beagle dogs. Cone beam CT images were used to create three dimensional images of the healed defects. Porous zirconia scaffolds were fabricated by milling custo...
Fig. 11. Energy dispersive spectrum of control Cp titanium specimen
Fig. 11. Energy dispersive spectrum of control Cp titanium specimen
Fig. 10. Energy dispersive spectrum of Cp titanium specimen coated with HA-Zn
Fig. 10. Energy dispersive spectrum of Cp titanium specimen coated with HA-Zn
Fig. 9. Scanning electron microphotograph of control Cp titanium specimen at ×20,000
Fig. 9. Scanning electron microphotograph of control Cp titanium specimen at ×20,000
Fig. 8. Scanning electron microphotograph of control Cp titanium specimen at ×10,000
Fig. 8. Scanning electron microphotograph of control Cp titanium specimen at ×10,000
Fig. 7. Scanning electron microphotograph of control Cp Titanium specimen at X 5,000
Fig. 7. Scanning electron microphotograph of control Cp Titanium specimen at X 5,000
Fig. 6. Scanning electron microphotograph of Cp titanium specimen coated with HA-Zn at ×20,000
Fig. 6. Scanning electron microphotograph of Cp titanium specimen coated with HA-Zn at ×20,000
Fig. 5. Scanning electron microphotograph of Cp Titanium specimen coated with HA-Zn at X10,000
Fig. 5. Scanning electron microphotograph of Cp Titanium specimen coated with HA-Zn at X10,000
Fig. 4. Scanning electron microphotograph of Cp titanium specimen coated with nano HA- Zn at ×5000
Fig. 4. Scanning electron microphotograph of Cp titanium specimen coated with nano HA- Zn at ×5000
Fig. 3. IR spectra of HA-Zn powder scrapped from coated titanium specimen
Fig. 3. IR spectra of HA-Zn powder scrapped from coated titanium specimen
Fig. 2. IR spectra of Ca(NO3)2·4 H2O powder prepared from a natural source (CB)
Fig. 2. IR spectra of Ca(NO3)2·4 H2O powder prepared from a natural source (CB)
Fig. 1. Graphical presentation of the electrochemical-deposition coating process’ equipment
Fig. 1. Graphical presentation of the electrochemical-deposition coating process’ equipment
Number of specimens
Mean ± (SD)
Standard error mean
F value
P value
Control
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El-Wassefy, N.A., Reicha, F.M. & Aref, N.S. Electro-chemical deposition of nano hydroxyapatite-zinc coating on titanium metal substrate.
Int J Implant Dent 3, 39 (2017). https://doi.org/10.1186/s40729-017-0095-1
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Received: 20 March 2017
Accepted: 28 July 2017
Published: 13 August 2017
DOI: https://doi.org/10.1186/s40729-017-0095-1
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
El-Wassefy N, Aref N, and Reicha F declare that they have no competing interests.
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Dental Biomaterials Department, Faculty of Dentistry, Mansoura University, 35516 El Gomhoria St., Mansoura, Egypt
N. A. El-Wassefy & N. S. Aref
Physics Department, Faculty of science, Mansoura University, 35516 El Gomhoria St., Mansoura, Egypt
F. M. Reicha
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The authors would like to express their gratitude for Dr. Sherif Kishk, Professor of Communication and Electrical Engineering, Faculty of Engineering, Mansoura University, for his help in photographing and analyzing the coating for adhesion test.
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Heydenrijk K, Meijer HJA, van der Reijden WA, Vissink A, Raghoebar GM, Stegenga B. Microbiota around root-formed endosseous implants. A review of the literature. October. ...
The electro-chemical method can be employed for HA-Zn coating deposition on titanium metal, where Ca source was a recycled cuttlebone fish to precipitate HA phases. Using a Zn anode on a low-sustained voltage was able to induce an even coat thickness of HA-Zn precipitation and increase the surface roughness significantly.
Yang et al. prepared a Zn-HA coating on Ti plates by an electrochemical process, and the SEM examination showed irregularly shaped rod-like crystals with hexagonal cross-section; this corresponded well with the current study results. They also concluded that a Zn-HA coating improves proliferation and differentiation of osteoblasts and would enhance implant osseointegration [11].
Ceramic coatings ...
Metallic orthopedic prosthesis is most commonly used due to its good mechanical properties, but its failure mostly occurs due to the lack of proper bone bonding and/or the occurrence of post-operative infections. Hydroxyapatite is commonly used as a bone filler biomaterial or as a coat for titanium prosthesis due to its decent biocompatibility, osseoconductivity, and bioactivity [26]. However, as ...
Following the examination of X cut areas after the adhesive tape removal; the adhesion was rated to be 5A, as no peeling or coat removal occurred along the incisions' length or at their intersection.
Figure 2 shows the FT-IR spectra of Ca(NO3)2·4H2O with weak sharp absorption peak bands at 742, 821, and 1048 cm−1, a strong broad absorption band at 1354 cm-1, and a strong shoulder absorption band at 1455 cm−1. A wide broad absorption band peak appears at 3442 cm−1 due to the presence of water. Figure 3 shows the FT-IR spectra of HA-Zn powder scrapped from CpTi specimens; the band a...
The coating was scrapped from Ti specimen's surface and investigated for its chemical structure using FT-IR spectroscopy. The powder was investigated by double-beam dispersive IR spectrometer (Nicolet iS10, Thermo Electron Corporation, UK) which utilized the selected range of 400 to 4000 wave numbers (cm−1) at 4 cm−1 resolution and averaging of 100 scans. Two milligrams of scrapped powder was...
Commercially pure Ti (CpTi) grade II specimens were cut down into plates with dimensions 10 × 10 × 2 mm and used as substrates (cathode material) for depositing HA and Zn. CpTi specimens were polished with successive grades of silicon carbide papers, ultra-sonicated in acetone (99.5%, EM Science), rinsed in distilled water, and then air dried at room temperature, before they were used f...
The aim of the present work was to develop well-adhered and uniform hydroxyapatite-zinc coatings on titanium metal substrate, through an in vitro electro-chemical deposition method. The coating was characterized for functional chemical group, surface morphology, surface chemical analysis, surface roughness, and coat adhesive bonding by Fourier transform infrared spectrometer (FT-IR), scanning elec...
The aim of the present work was to develop well-adhered and uniform hydroxyapatite-zinc coatings on titanium metal substrate, through an in vitro electro-chemical deposition method. The coating was characterized for functional chemical group, surface morphology, surface chemical analysis, surface roughness, and coat adhesive bonding by Fourier transform infrared spectrometer (FT-IR), scanning elec...
Titanium metal is one of the most widely used biomedical orthopedic materials because of its decent mechanical properties [1]. However, as an inert material, it cannot induce osteogenesis and has no antibacterial properties [2]. In order to improve surface bioactivity of titanium substrates, numerous methods have been proposed to cover it with bio-ceramic coatings [1]. Various clinical studies ...
Titanium is an inert metal that does not induce osteogenesis and has no antibacterial properties; it is proposed that hydroxyapatite coating can enhance its bioactivity, while zinc can contribute to antibacterial properties and improve osseointegration.
A nano-sized hydroxyapatite-zinc coating was deposited on commercially pure titanium using an electro-chemical process, in order to increase its ...
Figure 6. Cultivation and osteogenic differentiation of DFCs on PA after modification with collagen I. (Left) Relative cell number and (Right) normalized ALP activity.
Figure 6. Cultivation and osteogenic differentiation of DFCs on PA after modification with collagen I. (Left) Relative cell number and (Right) normalized ALP activity.
Figure 5. Evaluation of osteogenic differentiation. (A) Clustergram of PCR-array results; (B-C) histology of differentiated dental cells on AP (B) and SB (C). Representative results are shown for dNC-PCs.
Figure 5. Evaluation of osteogenic differentiation. (A) Clustergram of PCR-array results; (B-C) histology of differentiated dental cells on AP (B) and SB (C). Representative result...
Figure 4. Osteogenic differentiation of dental stem cells. Normalized ALP activity of dNC-PCs and DFCs on AP and SB (A) and on silicone (B). Cells were differentiated on standard cell culture dishes for control.
Figure 4. Osteogenic differentiation of dental stem cells. Normalized ALP activity of dNC-PCs and DFCs on AP and SB (A) and on silicone (B). Cells were differentiated on standar...
Figure 3. Evaluation of programmed cell death (apoptosis) in dental stem cells. (A) Flow cytometry analyses (for details materials and methods) show percentage of vital cells (black number), apoptotic cells (blue number), and dead cells (red number). (B) Western blot analyses show the expression of the pro-apoptotic marker BAX and the anti-apoptotic marker BCL2.
Figure 2. Cell proliferation of dNC-PCs and DFCs on tested materials. (A) and (B) Relative cell numbers; (C) spheroid cell clusters on silicone (representative pictures for DFCs); Silicone (24 and 48 h).
Figure 1. Cell attachment on tested materials. (A) Relative cell adherence of DFCs and dNC-PCs; (B) dental cells did little adhere on PA; representative pictures of DFCs.
In a previous study, we showed that TCP induced the programmed cell death (apoptosis) in DFCs. Our new study investigated therefore the induction of apoptosis in dental cells. While SB and soft materials did not induce apoptosis or cell death, AP induced obviously cell death and apoptosis in dental cells. Here, the results for dNC-PCs and DFCs were almost the same. Interestingly, neither sil...
Discussion
Scaffolds play an important role in tissue engineering. However, little is known about the proliferation and differentiation of DFCs and dNC-PCs on different types of materials. As we have learned from previous studies mechanical properties such as surface stiffness are decisive for a successful osteogenic differentiation of dental stem cells. Moreover, we showed that bone substi...
Results
Cell viability
Dental cells were cultivated in standard cell culture media until passage 6. Cell adherence and cell proliferation/growth were measured for the estimation of cell viability on tested rigid and soft materials. In Figure 1, the cell adherence of dNC-PCs on bone substitute materials was better than that of DFCs. However, both dental cells types adhered very well on silicone...
Cells positive for Caspase3/7 Green Detection Reagent were identified as apoptotic cells, while dead cells were positive for SYTOX® AADvanced dead cell stain. However, vital cells were negatively stained for both staining solutions.
Western blotting
For protein isolation, cells were treated with lysis buffer (250 μl phosphatase, 100 mM Na3VO4, 137 mM NaCl, 200 mM Tris, 480 mM NaF, 1% NP-4...
This incubation step with the implant material was repeated twice with fresh cell culture media. Three eluates were pooled for cell culture experiments. DFCs were seeded onto cell culture plates and cultivated in standard cell culture media. After cell seeding (12 to 24 h), cell culture media were changed, and cells were cultivated in cell culture media with material eluates. After 24 h of cultiva...
After washing with PBS, the gels were stored in PBS at 4°C. Before platting the cells, the gel was exposed to UV for 15 min for the sterilization and replace PBS with complete culture medium for 1 h at 37°C.
Implant materials
The bone substitutes Maxgraft® (AP) and Maxresorb® (SB) were obtained from the company Botiss (botiss dental GmbH, Berlin, Germany). Maxgraft® is a sterile, high-saf...
Methods
Cell culture
The isolation and characterization of DFCs and dNC-PCs were described in previous studies. DFCs were routinely cultivated in DMEM (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and 100 μg/ml penicillin/streptomycin (standard cell culture medium). dNC-PCs were cultivated in DMEM (Sigma-Aldrich) supplemented ...
Background
While bone substitute materials are routinely used, especially vertical bone, augmentation of the jaws is still a problematic step. Dental stem cells in combination with bone substitute materials may accelerate the augmentation of alveolar bone and perhaps, stem cell-based therapies can become an alternative to autologous, allogenic, or synthetic bone transplants and substitutes. How...
Abstract
Background
Dental stem cells in combination with implant materials may become an alternative to autologous bone transplants. For tissue engineering different types of soft and rigid implant materials are available, but little is known about the viability and the osteogenic differentiation of dental stem cells on these different types of materials. According to previous s...
Conclusions
The porous Col-HA composites developed in the present study are biocompatible and can be used as scaffolds for bone tissue regeneration. The Col-HA ratio is an important factor in promoting the attachment and proliferation of mouse MSCs. The Col-HA composite complexes have strong potentials in bone tissue regeneration applications. hPDSCs may be a suitable resource of cells for maxi...
Discussion
The findings of the presented study indicate that the porous sponge-like Col-HA composites have good biocompatibility and biomimetic properties and may be used as scaffolds for bone tissue regeneration. The Col-HA composites with ratios 80:20 and 50:50 supported the attachments and proliferations of mouse MSCs and hPDSCs. These findings indicate that Col-HA composite complexes have str...
Results
The sponge-like plugs of prototype Col-HA composites were successfully fabricated with different collagen and HA ratios. The macroscopic and SEM views of the prototype type I collagen without HA and 3 different ratios of collagen-HA (20%Col-80%HA; 50%Col-50%HA; 80%Col-20%HA) composites are shown in Figure 1. The SEM views show the inside microstructures of the prototype pure type I colla...
Materials and Methods
Synthesis of the Col-HA composites by direct precipitation in situ
Solutions of calcium salt and phosphoric acid (Ca/P = 1.66 mol) were used to synthesize HA particles and incorporate them on bovine type I collagen fibrils by a direct precipitation technique in situ. This technique was optimized to produce 3 different ratios of Col-HA composites (20%Col-80%HA; 50%Col-50%H...
Introduction
Combining a scaffold and living cells to form a tissue-engineering construct is an important concept for promoting the repair and regeneration of bone tissues. Mesenchymal stem cells are often used in such constructs due to their abilities to proliferate and differentiate toward bone-forming cells. The design and fabrication of scaffolds, stem cell isolation and characterization, and...
Abstract
Current bone grafting materials have significant limitations for repairing maxillofacial and dentoalveolar bone deficiencies. An ideal bone tissue-engineering construct is still lacking. The purpose of the present study was first to synthesize and develop a collagen-hydroxyapatite (Col-HA) composite through controlled in situ mineralization on type I collagen fibrils with nanometer-sized...
RESEARCH
Porous Collagen-Hydroxyapatite Scaffolds With Mesenchymal Stem Cells for Bone Regeneration
Li Ning, DDS, PhD , Hans Malmström, DDS , Yan-Fang Ren, DDS, MPH, PhD
Correspondence:
* Corresponding author, e-mail: yanfang_ren@urmc.rochester.edu
Article Citation:
Li Ning, Hans Malmström, Yan-Fang Ren, Porous Collagen-Hydroxyapatite Scaffolds With Mesench...