Endocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord Wharton’s jelly-derived mesenchymal stem cells
Xingxing Shi,* Kai Zhou,* Fei Huang,* Juan Zhang, Chen Wang Department of Prosthodontics, Jiangsu Key Laboratory of Oral Diseases, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, China *These authors contributed equally to this work Background: As a potentially bioactive...
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Dove Medical Press
2018
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oai:doaj.org-article:2d340a4498e94253b89e070c8c277d4b2021-12-02T08:08:25ZEndocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord Wharton’s jelly-derived mesenchymal stem cells1178-2013https://doaj.org/article/2d340a4498e94253b89e070c8c277d4b2018-03-01T00:00:00Zhttps://www.dovepress.com/endocytic-mechanisms-and-osteoinductive-profile-of-hydroxyapatite-nano-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Xingxing Shi,* Kai Zhou,* Fei Huang,* Juan Zhang, Chen Wang Department of Prosthodontics, Jiangsu Key Laboratory of Oral Diseases, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, China *These authors contributed equally to this work Background: As a potentially bioactive material, the widespread application of nanosized hydroxyapatite (nano-HAP) in the field of bone regeneration has increased the risk of human exposure. However, our understanding of the interaction between nano-HAP and stem cells implicated in bone repair remains incomplete. Methods: Here, we characterized the adhesion and cellular internalization of HAP nanoparticles (HANPs) with different sizes (20 nm np20 and 80 nm np80) and highlighted the involved pathway in their uptake using human umbilical cord Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs). In addition, the effects of HANPs on cell viability, apoptosis response, osteogenic differentiation, and underlying related mechanisms were explored. Results: It was shown that both types of HANPs readily adhered to the cellular membrane and were transported into the cells compared to micro-sized HAP particles (m-HAP; 12 µm). Interestingly, the endocytic routes of np20 and np80 differed, although they exhibited similar kinetics of adhesion and uptake. Our study revealed involvement of clathrin- and caveolin-mediated endocytosis as well as macropinocytosis in the np20 uptake. However, for np80, clathrin-mediated endocytosis and some as-yet-unidentified important uptake routes play central roles in their internalization. HANPs displayed a higher preference to accumulate in the cytoplasm compared to m-HAP, and HANPs were not detected in the nucleolus. Exposure to np20 for 24 h caused a decrease in cell viability, while cells completely recovered with an exposure time of 72 h. Furthermore, HANPs did not influence apoptosis and necrosis of hWJ-MSCs. Strikingly, HANPs enhanced mRNA levels of osteoblast-related genes and stimulated calcium mineral deposition, and this directly correlated with the activation in c-Jun N-terminal kinases and p38 pathways. Conclusion: Our data provide additional insight about the interactions of HANPs with MSCs and suggest their application potential in hard tissue regeneration. Keywords: HANPs, hWJ-MSCs, internalization, osteogenic differentiation, JNK, p38Shi XXZhou KHuang FZhang JWang CDove Medical PressarticleHANPshWJ-MSCsinternalizationosteogenic differentiationJNKp38Medicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 13, Pp 1457-1470 (2018) |
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HANPs hWJ-MSCs internalization osteogenic differentiation JNK p38 Medicine (General) R5-920 |
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HANPs hWJ-MSCs internalization osteogenic differentiation JNK p38 Medicine (General) R5-920 Shi XX Zhou K Huang F Zhang J Wang C Endocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord Wharton’s jelly-derived mesenchymal stem cells |
| description |
Xingxing Shi,* Kai Zhou,* Fei Huang,* Juan Zhang, Chen Wang Department of Prosthodontics, Jiangsu Key Laboratory of Oral Diseases, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, China *These authors contributed equally to this work Background: As a potentially bioactive material, the widespread application of nanosized hydroxyapatite (nano-HAP) in the field of bone regeneration has increased the risk of human exposure. However, our understanding of the interaction between nano-HAP and stem cells implicated in bone repair remains incomplete. Methods: Here, we characterized the adhesion and cellular internalization of HAP nanoparticles (HANPs) with different sizes (20 nm np20 and 80 nm np80) and highlighted the involved pathway in their uptake using human umbilical cord Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs). In addition, the effects of HANPs on cell viability, apoptosis response, osteogenic differentiation, and underlying related mechanisms were explored. Results: It was shown that both types of HANPs readily adhered to the cellular membrane and were transported into the cells compared to micro-sized HAP particles (m-HAP; 12 µm). Interestingly, the endocytic routes of np20 and np80 differed, although they exhibited similar kinetics of adhesion and uptake. Our study revealed involvement of clathrin- and caveolin-mediated endocytosis as well as macropinocytosis in the np20 uptake. However, for np80, clathrin-mediated endocytosis and some as-yet-unidentified important uptake routes play central roles in their internalization. HANPs displayed a higher preference to accumulate in the cytoplasm compared to m-HAP, and HANPs were not detected in the nucleolus. Exposure to np20 for 24 h caused a decrease in cell viability, while cells completely recovered with an exposure time of 72 h. Furthermore, HANPs did not influence apoptosis and necrosis of hWJ-MSCs. Strikingly, HANPs enhanced mRNA levels of osteoblast-related genes and stimulated calcium mineral deposition, and this directly correlated with the activation in c-Jun N-terminal kinases and p38 pathways. Conclusion: Our data provide additional insight about the interactions of HANPs with MSCs and suggest their application potential in hard tissue regeneration. Keywords: HANPs, hWJ-MSCs, internalization, osteogenic differentiation, JNK, p38 |
| format |
article |
| author |
Shi XX Zhou K Huang F Zhang J Wang C |
| author_facet |
Shi XX Zhou K Huang F Zhang J Wang C |
| author_sort |
Shi XX |
| title |
Endocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord Wharton’s jelly-derived mesenchymal stem cells |
| title_short |
Endocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord Wharton’s jelly-derived mesenchymal stem cells |
| title_full |
Endocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord Wharton’s jelly-derived mesenchymal stem cells |
| title_fullStr |
Endocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord Wharton’s jelly-derived mesenchymal stem cells |
| title_full_unstemmed |
Endocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord Wharton’s jelly-derived mesenchymal stem cells |
| title_sort |
endocytic mechanisms and osteoinductive profile of hydroxyapatite nanoparticles in human umbilical cord wharton’s jelly-derived mesenchymal stem cells |
| publisher |
Dove Medical Press |
| publishDate |
2018 |
| url |
https://doaj.org/article/2d340a4498e94253b89e070c8c277d4b |
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