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Preparation of laponite bioceramics for potential bone tissue engineering applications.

Wang C, Wang S, Li K, Ju Y, Li J, Zhang Y, Li J, Liu X, Shi X, Zhao Q - PLoS ONE (2014)

Bottom Line: Our results reveal that the LAP bioceramic possesses an excellent surface hydrophilicity and serum absorption capacity, and good cytocompatibility and hemocompatibility as demonstrated by resazurin reduction assay of rat mesenchymal stem cells (rMSCs) and hemolytic assay of pig red blood cells, respectively.Most strikingly, alkaline phosphatase activity together with alizarin red staining results reveal that the produced LAP bioceramic is able to induce osteoblast differentiation of rMSCs in growth medium without any inducing factors.Finally, in vivo animal implantation, acute systemic toxicity test and hematoxylin and eosin (H&E)-staining data demonstrate that the prepared LAP bioceramic displays an excellent biosafety and is able to heal the bone defect.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedics, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, P. R. China.

ABSTRACT
We report a facile approach to preparing laponite (LAP) bioceramics via sintering LAP powder compacts for bone tissue engineering applications. The sintering behavior and mechanical properties of LAP compacts under different temperatures, heating rates, and soaking times were investigated. We show that LAP bioceramic with a smooth and porous surface can be formed at 800°C with a heating rate of 5°C/h for 6 h under air. The formed LAP bioceramic was systematically characterized via different methods. Our results reveal that the LAP bioceramic possesses an excellent surface hydrophilicity and serum absorption capacity, and good cytocompatibility and hemocompatibility as demonstrated by resazurin reduction assay of rat mesenchymal stem cells (rMSCs) and hemolytic assay of pig red blood cells, respectively. The potential bone tissue engineering applicability of LAP bioceramic was explored by studying the surface mineralization behavior via soaking in simulated body fluid (SBF), as well as the surface cellular response of rMSCs. Our results suggest that LAP bioceramic is able to induce hydroxyapatite deposition on its surface when soaked in SBF and rMSCs can proliferate well on the LAP bioceramic surface. Most strikingly, alkaline phosphatase activity together with alizarin red staining results reveal that the produced LAP bioceramic is able to induce osteoblast differentiation of rMSCs in growth medium without any inducing factors. Finally, in vivo animal implantation, acute systemic toxicity test and hematoxylin and eosin (H&E)-staining data demonstrate that the prepared LAP bioceramic displays an excellent biosafety and is able to heal the bone defect. Findings from this study suggest that the developed LAP bioceramic holds a great promise for treating bone defects in bone tissue engineering.

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Protein adsorption and metabolic activity assay of rMSCs.(a) The adsorption of protein onto TCP and LAP ceramic (mean ± S.D., n = 3). (b) and (c) show the SEM micrographs of the TCP and LAP ceramic with protein adsorption, respectively. (d) shows the metabolic activity assay of rMSCs cultured onto TCP and LAP ceramic (mean ± S.D., n = 3). (e) shows the micrograph of rMSCs proliferated onto the LAP bioceramic for 14 days. (f) is the magnified image of (e).
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pone-0099585-g006: Protein adsorption and metabolic activity assay of rMSCs.(a) The adsorption of protein onto TCP and LAP ceramic (mean ± S.D., n = 3). (b) and (c) show the SEM micrographs of the TCP and LAP ceramic with protein adsorption, respectively. (d) shows the metabolic activity assay of rMSCs cultured onto TCP and LAP ceramic (mean ± S.D., n = 3). (e) shows the micrograph of rMSCs proliferated onto the LAP bioceramic for 14 days. (f) is the magnified image of (e).

Mentions: An ideal material for bone tissue engineering applications should also have the ability to absorb protein onto its surface, thus providing enough nutrition for cell growth and migration [35]. Based on this point, we then explored the serum (FBS) adsorption capacity of LAP bioceramic (Figures 6a–c). As shown in Figure 6a, the LAP bioceramic can absorb similar amount of FBS when compared to TCP after 24 h incubation (9.1±0.9 mg/well versus 8.4±0.2 mg/well, p>0.05). The adsorbed FBS was further confirmed by SEM (Figures 6b–c), where solid-state FBS is attached onto the surfaces of both TCP and LAP bioceramic. Since TCP has been already coated with other materials to enhance cell attachment and proliferation, it is reasonable to conclude that the protein adsorption onto LAP bioceramic surface may induce enhanced cellular response, similar to our previous reports [28], [36].


Preparation of laponite bioceramics for potential bone tissue engineering applications.

Wang C, Wang S, Li K, Ju Y, Li J, Zhang Y, Li J, Liu X, Shi X, Zhao Q - PLoS ONE (2014)

Protein adsorption and metabolic activity assay of rMSCs.(a) The adsorption of protein onto TCP and LAP ceramic (mean ± S.D., n = 3). (b) and (c) show the SEM micrographs of the TCP and LAP ceramic with protein adsorption, respectively. (d) shows the metabolic activity assay of rMSCs cultured onto TCP and LAP ceramic (mean ± S.D., n = 3). (e) shows the micrograph of rMSCs proliferated onto the LAP bioceramic for 14 days. (f) is the magnified image of (e).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4067276&req=5

pone-0099585-g006: Protein adsorption and metabolic activity assay of rMSCs.(a) The adsorption of protein onto TCP and LAP ceramic (mean ± S.D., n = 3). (b) and (c) show the SEM micrographs of the TCP and LAP ceramic with protein adsorption, respectively. (d) shows the metabolic activity assay of rMSCs cultured onto TCP and LAP ceramic (mean ± S.D., n = 3). (e) shows the micrograph of rMSCs proliferated onto the LAP bioceramic for 14 days. (f) is the magnified image of (e).
Mentions: An ideal material for bone tissue engineering applications should also have the ability to absorb protein onto its surface, thus providing enough nutrition for cell growth and migration [35]. Based on this point, we then explored the serum (FBS) adsorption capacity of LAP bioceramic (Figures 6a–c). As shown in Figure 6a, the LAP bioceramic can absorb similar amount of FBS when compared to TCP after 24 h incubation (9.1±0.9 mg/well versus 8.4±0.2 mg/well, p>0.05). The adsorbed FBS was further confirmed by SEM (Figures 6b–c), where solid-state FBS is attached onto the surfaces of both TCP and LAP bioceramic. Since TCP has been already coated with other materials to enhance cell attachment and proliferation, it is reasonable to conclude that the protein adsorption onto LAP bioceramic surface may induce enhanced cellular response, similar to our previous reports [28], [36].

Bottom Line: Our results reveal that the LAP bioceramic possesses an excellent surface hydrophilicity and serum absorption capacity, and good cytocompatibility and hemocompatibility as demonstrated by resazurin reduction assay of rat mesenchymal stem cells (rMSCs) and hemolytic assay of pig red blood cells, respectively.Most strikingly, alkaline phosphatase activity together with alizarin red staining results reveal that the produced LAP bioceramic is able to induce osteoblast differentiation of rMSCs in growth medium without any inducing factors.Finally, in vivo animal implantation, acute systemic toxicity test and hematoxylin and eosin (H&E)-staining data demonstrate that the prepared LAP bioceramic displays an excellent biosafety and is able to heal the bone defect.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedics, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, P. R. China.

ABSTRACT
We report a facile approach to preparing laponite (LAP) bioceramics via sintering LAP powder compacts for bone tissue engineering applications. The sintering behavior and mechanical properties of LAP compacts under different temperatures, heating rates, and soaking times were investigated. We show that LAP bioceramic with a smooth and porous surface can be formed at 800°C with a heating rate of 5°C/h for 6 h under air. The formed LAP bioceramic was systematically characterized via different methods. Our results reveal that the LAP bioceramic possesses an excellent surface hydrophilicity and serum absorption capacity, and good cytocompatibility and hemocompatibility as demonstrated by resazurin reduction assay of rat mesenchymal stem cells (rMSCs) and hemolytic assay of pig red blood cells, respectively. The potential bone tissue engineering applicability of LAP bioceramic was explored by studying the surface mineralization behavior via soaking in simulated body fluid (SBF), as well as the surface cellular response of rMSCs. Our results suggest that LAP bioceramic is able to induce hydroxyapatite deposition on its surface when soaked in SBF and rMSCs can proliferate well on the LAP bioceramic surface. Most strikingly, alkaline phosphatase activity together with alizarin red staining results reveal that the produced LAP bioceramic is able to induce osteoblast differentiation of rMSCs in growth medium without any inducing factors. Finally, in vivo animal implantation, acute systemic toxicity test and hematoxylin and eosin (H&E)-staining data demonstrate that the prepared LAP bioceramic displays an excellent biosafety and is able to heal the bone defect. Findings from this study suggest that the developed LAP bioceramic holds a great promise for treating bone defects in bone tissue engineering.

Show MeSH
Related in: MedlinePlus