Chitosan nanoparticles for sustained release of metformin and its derived synthetic biopolymer for bone regeneration
Background: There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for treating them. Chitosan is a natural biopolymer; it is used in approaches for sustained slow release and osteogenesis, and metformin has osteoinduct...
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| Published in | Frontiers in bioengineering and biotechnology Vol. 11; p. 1169496 |
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| Main Authors | , , , , , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
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Switzerland
Frontiers Media S.A
05.07.2023
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| Abstract | Background:
There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for treating them. Chitosan is a natural biopolymer; it is used in approaches for sustained slow release and osteogenesis, and metformin has osteoinductivity. Our study aimed to synthesize chitosan and human serum albumin (HSA) with a metformin nanoformulation to evaluate the therapeutic effects of this nanoformulation on bone defects
in vitro
.
Methods:
A pluripotent differentiation assay was performed
in vitro
on mouse bone marrow mesenchymal stem cells (BMSCs). Cell Counting Kit-8 was used to detect whether metformin was toxic to BMSCs. The osteogenesis-related gene expression of osteocalcin (OCN) and osteoprotegerin (OPG) from BMSCs was tested by real-time polymerase chain reaction (PCR). HSA, metformin hydrochloride, and chitosan mixtures were magnetically stirred to finish the assembly of metformin/HSA/chitosan nanoparticles (MHC NPs). The MHC NPs were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and Fourier transform infrared spectroscopy (FT-IR). To test the expression of OCN and OPG, western blot were used. MHC NPs were evaluated
in vitro
for their osteoinductivity using alkaline phosphatase (ALP).
Results:
BMSCs successfully differentiated into osteogenic and adipogenic lineages
in vitro
. According to real-time PCR, a 50 µM concentration of metformin promoted osteogenesis in BMSCs most effectively by upregulating the osteogenic markers OCN and OPG. The microstructure of MHC NPs was spherical with an average nanosize of 20 ± 4.7 nm and zeta potential of −8.3 mV. A blueshift and redshift were observed in MHC NPs following exposure to wavelengths of 1,600–1,900 and 2,000–3,700 nm, respectively. The encapsulation (%) of metformin was more than 90%. The simulation study showed that MHC NPs have good stability and it could release metformin slowly
in vitro
at room temperature. Upon treatment with the studied MHC NPs for 3 days, ALP was significantly elevated in BMSCs. In addition, the MHC NPs-treated BMSCs upregulated the expression of OPG and OCN, as shown by real-time PCR and western blot.
Conclusion:
MHC NPs have a stable metformin release effect and osteogenic ability. Therefore, as a derived synthetic biopolymer, it is expected to play a role in bone tissue regeneration. |
|---|---|
| AbstractList | Background:
There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for treating them. Chitosan is a natural biopolymer; it is used in approaches for sustained slow release and osteogenesis, and metformin has osteoinductivity. Our study aimed to synthesize chitosan and human serum albumin (HSA) with a metformin nanoformulation to evaluate the therapeutic effects of this nanoformulation on bone defects
in vitro
.
Methods:
A pluripotent differentiation assay was performed
in vitro
on mouse bone marrow mesenchymal stem cells (BMSCs). Cell Counting Kit-8 was used to detect whether metformin was toxic to BMSCs. The osteogenesis-related gene expression of osteocalcin (OCN) and osteoprotegerin (OPG) from BMSCs was tested by real-time polymerase chain reaction (PCR). HSA, metformin hydrochloride, and chitosan mixtures were magnetically stirred to finish the assembly of metformin/HSA/chitosan nanoparticles (MHC NPs). The MHC NPs were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and Fourier transform infrared spectroscopy (FT-IR). To test the expression of OCN and OPG, western blot were used. MHC NPs were evaluated
in vitro
for their osteoinductivity using alkaline phosphatase (ALP).
Results:
BMSCs successfully differentiated into osteogenic and adipogenic lineages
in vitro
. According to real-time PCR, a 50 µM concentration of metformin promoted osteogenesis in BMSCs most effectively by upregulating the osteogenic markers OCN and OPG. The microstructure of MHC NPs was spherical with an average nanosize of 20 ± 4.7 nm and zeta potential of −8.3 mV. A blueshift and redshift were observed in MHC NPs following exposure to wavelengths of 1,600–1,900 and 2,000–3,700 nm, respectively. The encapsulation (%) of metformin was more than 90%. The simulation study showed that MHC NPs have good stability and it could release metformin slowly
in vitro
at room temperature. Upon treatment with the studied MHC NPs for 3 days, ALP was significantly elevated in BMSCs. In addition, the MHC NPs-treated BMSCs upregulated the expression of OPG and OCN, as shown by real-time PCR and western blot.
Conclusion:
MHC NPs have a stable metformin release effect and osteogenic ability. Therefore, as a derived synthetic biopolymer, it is expected to play a role in bone tissue regeneration. There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for treating them. Chitosan is a natural biopolymer; it is used in approaches for sustained slow release and osteogenesis, and metformin has osteoinductivity. Our study aimed to synthesize chitosan and human serum albumin (HSA) with a metformin nanoformulation to evaluate the therapeutic effects of this nanoformulation on bone defects . A pluripotent differentiation assay was performed on mouse bone marrow mesenchymal stem cells (BMSCs). Cell Counting Kit-8 was used to detect whether metformin was toxic to BMSCs. The osteogenesis-related gene expression of osteocalcin (OCN) and osteoprotegerin (OPG) from BMSCs was tested by real-time polymerase chain reaction (PCR). HSA, metformin hydrochloride, and chitosan mixtures were magnetically stirred to finish the assembly of metformin/HSA/chitosan nanoparticles (MHC NPs). The MHC NPs were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and Fourier transform infrared spectroscopy (FT-IR). To test the expression of OCN and OPG, western blot were used. MHC NPs were evaluated for their osteoinductivity using alkaline phosphatase (ALP). BMSCs successfully differentiated into osteogenic and adipogenic lineages . According to real-time PCR, a 50 µM concentration of metformin promoted osteogenesis in BMSCs most effectively by upregulating the osteogenic markers OCN and OPG. The microstructure of MHC NPs was spherical with an average nanosize of 20 ± 4.7 nm and zeta potential of -8.3 mV. A blueshift and redshift were observed in MHC NPs following exposure to wavelengths of 1,600-1,900 and 2,000-3,700 nm, respectively. The encapsulation (%) of metformin was more than 90%. The simulation study showed that MHC NPs have good stability and it could release metformin slowly at room temperature. Upon treatment with the studied MHC NPs for 3 days, ALP was significantly elevated in BMSCs. In addition, the MHC NPs-treated BMSCs upregulated the expression of OPG and OCN, as shown by real-time PCR and western blot. MHC NPs have a stable metformin release effect and osteogenic ability. Therefore, as a derived synthetic biopolymer, it is expected to play a role in bone tissue regeneration. Background: There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for treating them. Chitosan is a natural biopolymer; it is used in approaches for sustained slow release and osteogenesis, and metformin has osteoinductivity. Our study aimed to synthesize chitosan and human serum albumin (HSA) with a metformin nanoformulation to evaluate the therapeutic effects of this nanoformulation on bone defects in vitro.Methods: A pluripotent differentiation assay was performed in vitro on mouse bone marrow mesenchymal stem cells (BMSCs). Cell Counting Kit-8 was used to detect whether metformin was toxic to BMSCs. The osteogenesis-related gene expression of osteocalcin (OCN) and osteoprotegerin (OPG) from BMSCs was tested by real-time polymerase chain reaction (PCR). HSA, metformin hydrochloride, and chitosan mixtures were magnetically stirred to finish the assembly of metformin/HSA/chitosan nanoparticles (MHC NPs). The MHC NPs were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and Fourier transform infrared spectroscopy (FT-IR). To test the expression of OCN and OPG, western blot were used. MHC NPs were evaluated in vitro for their osteoinductivity using alkaline phosphatase (ALP).Results: BMSCs successfully differentiated into osteogenic and adipogenic lineages in vitro. According to real-time PCR, a 50 µM concentration of metformin promoted osteogenesis in BMSCs most effectively by upregulating the osteogenic markers OCN and OPG. The microstructure of MHC NPs was spherical with an average nanosize of 20 ± 4.7 nm and zeta potential of −8.3 mV. A blueshift and redshift were observed in MHC NPs following exposure to wavelengths of 1,600–1,900 and 2,000–3,700 nm, respectively. The encapsulation (%) of metformin was more than 90%. The simulation study showed that MHC NPs have good stability and it could release metformin slowly in vitro at room temperature. Upon treatment with the studied MHC NPs for 3 days, ALP was significantly elevated in BMSCs. In addition, the MHC NPs-treated BMSCs upregulated the expression of OPG and OCN, as shown by real-time PCR and western blot.Conclusion: MHC NPs have a stable metformin release effect and osteogenic ability. Therefore, as a derived synthetic biopolymer, it is expected to play a role in bone tissue regeneration. Background: There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for treating them. Chitosan is a natural biopolymer; it is used in approaches for sustained slow release and osteogenesis, and metformin has osteoinductivity. Our study aimed to synthesize chitosan and human serum albumin (HSA) with a metformin nanoformulation to evaluate the therapeutic effects of this nanoformulation on bone defects in vitro. Methods: A pluripotent differentiation assay was performed in vitro on mouse bone marrow mesenchymal stem cells (BMSCs). Cell Counting Kit-8 was used to detect whether metformin was toxic to BMSCs. The osteogenesis-related gene expression of osteocalcin (OCN) and osteoprotegerin (OPG) from BMSCs was tested by real-time polymerase chain reaction (PCR). HSA, metformin hydrochloride, and chitosan mixtures were magnetically stirred to finish the assembly of metformin/HSA/chitosan nanoparticles (MHC NPs). The MHC NPs were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and Fourier transform infrared spectroscopy (FT-IR). To test the expression of OCN and OPG, western blot were used. MHC NPs were evaluated in vitro for their osteoinductivity using alkaline phosphatase (ALP). Results: BMSCs successfully differentiated into osteogenic and adipogenic lineages in vitro. According to real-time PCR, a 50 µM concentration of metformin promoted osteogenesis in BMSCs most effectively by upregulating the osteogenic markers OCN and OPG. The microstructure of MHC NPs was spherical with an average nanosize of 20 ± 4.7 nm and zeta potential of -8.3 mV. A blueshift and redshift were observed in MHC NPs following exposure to wavelengths of 1,600-1,900 and 2,000-3,700 nm, respectively. The encapsulation (%) of metformin was more than 90%. The simulation study showed that MHC NPs have good stability and it could release metformin slowly in vitro at room temperature. Upon treatment with the studied MHC NPs for 3 days, ALP was significantly elevated in BMSCs. In addition, the MHC NPs-treated BMSCs upregulated the expression of OPG and OCN, as shown by real-time PCR and western blot. Conclusion: MHC NPs have a stable metformin release effect and osteogenic ability. Therefore, as a derived synthetic biopolymer, it is expected to play a role in bone tissue regeneration.Background: There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for treating them. Chitosan is a natural biopolymer; it is used in approaches for sustained slow release and osteogenesis, and metformin has osteoinductivity. Our study aimed to synthesize chitosan and human serum albumin (HSA) with a metformin nanoformulation to evaluate the therapeutic effects of this nanoformulation on bone defects in vitro. Methods: A pluripotent differentiation assay was performed in vitro on mouse bone marrow mesenchymal stem cells (BMSCs). Cell Counting Kit-8 was used to detect whether metformin was toxic to BMSCs. The osteogenesis-related gene expression of osteocalcin (OCN) and osteoprotegerin (OPG) from BMSCs was tested by real-time polymerase chain reaction (PCR). HSA, metformin hydrochloride, and chitosan mixtures were magnetically stirred to finish the assembly of metformin/HSA/chitosan nanoparticles (MHC NPs). The MHC NPs were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and Fourier transform infrared spectroscopy (FT-IR). To test the expression of OCN and OPG, western blot were used. MHC NPs were evaluated in vitro for their osteoinductivity using alkaline phosphatase (ALP). Results: BMSCs successfully differentiated into osteogenic and adipogenic lineages in vitro. According to real-time PCR, a 50 µM concentration of metformin promoted osteogenesis in BMSCs most effectively by upregulating the osteogenic markers OCN and OPG. The microstructure of MHC NPs was spherical with an average nanosize of 20 ± 4.7 nm and zeta potential of -8.3 mV. A blueshift and redshift were observed in MHC NPs following exposure to wavelengths of 1,600-1,900 and 2,000-3,700 nm, respectively. The encapsulation (%) of metformin was more than 90%. The simulation study showed that MHC NPs have good stability and it could release metformin slowly in vitro at room temperature. Upon treatment with the studied MHC NPs for 3 days, ALP was significantly elevated in BMSCs. In addition, the MHC NPs-treated BMSCs upregulated the expression of OPG and OCN, as shown by real-time PCR and western blot. Conclusion: MHC NPs have a stable metformin release effect and osteogenic ability. Therefore, as a derived synthetic biopolymer, it is expected to play a role in bone tissue regeneration. |
| Author | Su, Xiao-Lin Feng, Yao Dusenge, Marie Aimee Tan, Li Guo, Yue Chen, Ning-Xin Ye, Qin Zhang, Qian Feng, Yun-Zhi Hu, Jing Ou-Yang, Ze-Yue Zhao, Ya-Qiong Liu, Qiong Chen, Yun Zhong, Meng-Mei Zhao, Jie Peng, Yong-Bo Yuan, Hui |
| AuthorAffiliation | 1 Department of Stomatology , The Second Xiangya Hospital , Central South University , Changsha , Hunan , China 2 Chongqing Key Laboratory for Pharmaceutical Metabolism Research , The Key Laboratory of Biochemistry and Molecular Pharmacology , College of Pharmacy , Chongqing Medical University , Chongqing , China |
| AuthorAffiliation_xml | – name: 1 Department of Stomatology , The Second Xiangya Hospital , Central South University , Changsha , Hunan , China – name: 2 Chongqing Key Laboratory for Pharmaceutical Metabolism Research , The Key Laboratory of Biochemistry and Molecular Pharmacology , College of Pharmacy , Chongqing Medical University , Chongqing , China |
| Author_xml | – sequence: 1 givenname: Ning-Xin surname: Chen fullname: Chen, Ning-Xin – sequence: 2 givenname: Xiao-Lin surname: Su fullname: Su, Xiao-Lin – sequence: 3 givenname: Yao surname: Feng fullname: Feng, Yao – sequence: 4 givenname: Qiong surname: Liu fullname: Liu, Qiong – sequence: 5 givenname: Li surname: Tan fullname: Tan, Li – sequence: 6 givenname: Hui surname: Yuan fullname: Yuan, Hui – sequence: 7 givenname: Yun surname: Chen fullname: Chen, Yun – sequence: 8 givenname: Jie surname: Zhao fullname: Zhao, Jie – sequence: 9 givenname: Ya-Qiong surname: Zhao fullname: Zhao, Ya-Qiong – sequence: 10 givenname: Marie Aimee surname: Dusenge fullname: Dusenge, Marie Aimee – sequence: 11 givenname: Jing surname: Hu fullname: Hu, Jing – sequence: 12 givenname: Qin surname: Ye fullname: Ye, Qin – sequence: 13 givenname: Ze-Yue surname: Ou-Yang fullname: Ou-Yang, Ze-Yue – sequence: 14 givenname: Meng-Mei surname: Zhong fullname: Zhong, Meng-Mei – sequence: 15 givenname: Qian surname: Zhang fullname: Zhang, Qian – sequence: 16 givenname: Yue surname: Guo fullname: Guo, Yue – sequence: 17 givenname: Yun-Zhi surname: Feng fullname: Feng, Yun-Zhi – sequence: 18 givenname: Yong-Bo surname: Peng fullname: Peng, Yong-Bo |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/37476483$$D View this record in MEDLINE/PubMed |
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| ContentType | Journal Article |
| Copyright | Copyright © 2023 Chen, Su, Feng, Liu, Tan, Yuan, Chen, Zhao, Zhao, Dusenge, Hu, Ye, Ou-Yang, Zhong, Zhang, Guo, Feng and Peng. Copyright © 2023 Chen, Su, Feng, Liu, Tan, Yuan, Chen, Zhao, Zhao, Dusenge, Hu, Ye, Ou-Yang, Zhong, Zhang, Guo, Feng and Peng. 2023 Chen, Su, Feng, Liu, Tan, Yuan, Chen, Zhao, Zhao, Dusenge, Hu, Ye, Ou-Yang, Zhong, Zhang, Guo, Feng and Peng |
| Copyright_xml | – notice: Copyright © 2023 Chen, Su, Feng, Liu, Tan, Yuan, Chen, Zhao, Zhao, Dusenge, Hu, Ye, Ou-Yang, Zhong, Zhang, Guo, Feng and Peng. – notice: Copyright © 2023 Chen, Su, Feng, Liu, Tan, Yuan, Chen, Zhao, Zhao, Dusenge, Hu, Ye, Ou-Yang, Zhong, Zhang, Guo, Feng and Peng. 2023 Chen, Su, Feng, Liu, Tan, Yuan, Chen, Zhao, Zhao, Dusenge, Hu, Ye, Ou-Yang, Zhong, Zhang, Guo, Feng and Peng |
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| Keywords | chitosan biopolymer bone regeneration metformin drug release BMSCs |
| Language | English |
| License | Copyright © 2023 Chen, Su, Feng, Liu, Tan, Yuan, Chen, Zhao, Zhao, Dusenge, Hu, Ye, Ou-Yang, Zhong, Zhang, Guo, Feng and Peng. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 J. Blas Pagador, Jesús Usón Minimally Invasive Surgery Center, Spain Edited by: Dingpei Long, Georgia State University, United States Reviewed by: Hai Zhang, University of Washington, United States These authors have contributed equally to this work and share first authorship |
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| References | Chen (B8) 2022; 18 Sudhakar (B36) 2020; 150 Liu (B20) 2022; 10 Xu (B41) 2021; 338 Zhu (B43) 2021; 21 De Jong (B10) 2008; 3 Lee (B19) 2009; 19 Ma (B23) 2016; 54 Buck (B6) 2012; 129 Galindo-Moreno (B13) 2022; 33 Peers (B30) 2020; 326 Guo (B15) 2016; 30 Lv (B21) 2020; 11 Ortiz (B29) 2022; 199 Mabrouk (B24) 2019; 107 Weitzmann (B40) 2015; 11 MacNeil (B25) 2020; 16 Fakhri (B12) 2020; 162 Tan (B37) 2021; 13 Qian (B33) 2019; 11 Zhao (B42) 2020; 14 Ma (B22) 2018; 22 Peng (B31) 2017; 56 Bahrambeigi (B3) 2019; 109 Tang (B38) 2016; 83 Majidinia (B26) 2018; 233 Myeroff (B27) 2011; 93 Qasim (B32) 2019; 14 Ren (B35) 2021; 10 Oliveira É (B28) 2021; 22 Khan (B17) 2005; 13 Chinnaiyan (B9) 2019; 125 Bhattarai (B4) 2006; 1 Dimitriou (B11) 2011; 9 Gulati (B14) 2012; 8 Khodadadi Yazdi (B18) 2020; 326 Adamička (B1) 2021; 2021 Aguilar (B2) 2019; 24 Bhujbal (B5) 2018; 19 Cai (B7) 2022; 23 Hashemi (B16) 2020; 15 Wang (B39) 2021; 329 Qiao (B34) 2011; 11 |
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There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for... There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for treating them.... Background: There are considerable socioeconomic costs associated with bone defects, making regenerative medicine an increasingly attractive option for... |
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| Title | Chitosan nanoparticles for sustained release of metformin and its derived synthetic biopolymer for bone regeneration |
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