31 results on '"An, Guangpeng"'
Search Results
2. The effect of fiber size and pore size on cell proliferation and infiltration in PLLA scaffolds on bone tissue engineering
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Guangpeng Cui, Guojun Song, Wenhua Zhao, Xuejun Wang, Chunyao Li, and Tao Lou
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Pore size ,Scaffold ,Materials science ,Polyesters ,Nanofibers ,Biomedical Engineering ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Cell Line ,Biomaterials ,Tissue engineering ,medicine ,Humans ,Cell Proliferation ,Osteoblasts ,Tissue Engineering ,Tissue Scaffolds ,Macropore ,Cell growth ,021001 nanoscience & nanotechnology ,medicine.disease ,Microstructure ,0104 chemical sciences ,0210 nano-technology ,Porosity ,Infiltration (medical) ,Protein adsorption ,Biomedical engineering - Abstract
The scaffold microstructure has a great impact on cell functions in tissue engineering. Herein, the PLLA scaffolds with hierarchical fiber size and pore size were successfully fabricated by thermal-induced phase separation or combined thermal-induced phase separation and salt leaching methods. The PLLA scaffolds were fabricated as microfibrous scaffolds, microfibrous scaffolds with macropores (50–350 µm), nanofibrous scaffolds with micropores (100 nm to 10 µm), and nanofibrous scaffolds with both macropores and micropores by tailoring selective solvents for forming different fiber size and pre-sieved salts for creating controlled pore size. Among the four kinds of PLLA scaffolds, the nanofibrous scaffolds with both macropores and micropores provided a favorable microenvironment for protein adsorption, cell proliferation, and cell infiltration. The results further confirmed the significance of fiber size and pore size on the biological properties, and a scaffold with both micropores and macropores, and a nanofibrous matrix might have promising applications in bone tissue engineering.
- Published
- 2016
3. Eco-friendly strategy for synthesizing europium-complex-based hybrid materials using bifunctional hydroxyapatite.
- Author
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Jiang, Guangpeng, Ma, Yuheng, Zhang, Sihua, Chen, Rulin, Li, Zhe, Li, Jun, Li, Ruiqian, and Tao, Dongliang
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HYBRID materials , *HYDROXYAPATITE , *ARTIFICIAL bones , *RARE earth metals , *TISSUE engineering , *EUROPIUM , *CHALK - Abstract
[Display omitted] • Eu-complex-based hybrids were prepared using hydroxyapatite as both eco-friendly alkaline reagent and matrix. • The quantum yield and lifetime of the hybrid materials are comparable to that of pure europium complex. • The hybrid materials can be used in various fields, including artificial bone, thermometer, and luminescent chalks. • The green strategy is applicable to Tb-complexes-based hybrid materials. Synthesizing lanthanide complexes and their hybrid materials often involves the utilization of hazard base reagents to neutralize the protons generated in coordination reactions. Here, we propose for the first time a green and facile route to synthesize europium-complex-based hybrid materials by using micrometer-sized hydroxyapatite (HAP). Simultaneous formations of the europium complex and monetite took place after the reaction. Not only did HAP function as an eco-friendly alkaline reagent, but it also acted as a matrix to disperse the europium complexes. The quantum yield and lifetime of the as-synthesized hybrid materials are comparable to that of pure europium complex prepared by using traditional base reagents. Furthermore, versatile applications of the hybrid materials are also demonstrated in our proof-of-concept experiments, including bone tissue engineering, luminescent toys, and portable luminescent chalks. Notably, the proposed eco-friendly strategy is also applicable to other types of lanthanide-complexes-based hybrid materials. [ABSTRACT FROM AUTHOR]
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- 2023
- Full Text
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4. Evaluation of the viability and osteogenic differentiation of cryopreserved human adipose-derived stem cells
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Yilin Cao, Yulin Li, Lei Cui, Guangpeng Liu, Wei Liu, Heng Zhou, and Gang Li
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Adult ,Cell Survival ,Cellular differentiation ,Osteocalcin ,Adipose tissue ,General Biochemistry, Genetics and Molecular Biology ,Cryopreservation ,Flow cytometry ,Tissue engineering ,Adipocytes ,medicine ,Humans ,Dimethyl Sulfoxide ,Cells, Cultured ,Osteoblasts ,biology ,medicine.diagnostic_test ,Stem Cells ,Cell Differentiation ,General Medicine ,Middle Aged ,Flow Cytometry ,Molecular biology ,Cell biology ,Adipose Tissue ,biology.protein ,Alkaline phosphatase ,Female ,Stem cell ,General Agricultural and Biological Sciences - Abstract
Human adipose-derived stem cells (ASCs) have the ability to differentiate into osteoblasts and thus the potential therapeutic use to tissue-engineer bone, so a reliable method for cell storage is necessary. The aim of this study was to determine whether a simple method of cryopreservation with 10% Me(2)SO as a protectant had an effect on proliferation potential and osteogenic differentiation of ASCs isolated from fresh human adipose tissue. ASCs were harvested from 6 human lipoaspirates and each was halved for either cryopreservation in liquid nitrogen for 2 weeks or for control culture. Cells from the second-passage were plated at a density of 5000cells/well in 24-well plates and cultured with or without osteogenic media for 14 days. Cell surface antigens were used to identify the cryopreserved ASCs by flow cytometry. The proliferation rate of both populations was evaluated using a cell DNA assay. To detect osteogenic differentiation of both the cryopreserved and non-cryopreserved populations, determination of osteoblastic protein production (alkaline phosphatase and osteocalcin) and excellular matrix calcification (calcium content) was applied. The expression of osteoblastic-associated genes was also analyzed using reverse-transcription polymerase chain reaction. These results demonstrate that cryopreservation has no effect on the phenotype, proliferation or osteogenic differentiation of human ASCs, showing cryopreserved human ASCs might be applied for bone tissue engineering.
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- 2008
5. Tissue-engineered bone formation using human bone marrow stromal cells and novel β-tricalcium phosphate
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Guangpeng Liu, Li Zhao, Wei Liu, Yilin Cao, and Lei Cui
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Calcium Phosphates ,Stromal cell ,Materials science ,Biomedical Engineering ,Mice, Nude ,Connective tissue ,Bioengineering ,Biomaterials ,Mice ,Osteogenesis ,In vivo ,Materials Testing ,medicine ,Animals ,Humans ,MTT assay ,Cells, Cultured ,Cell Proliferation ,Mice, Inbred BALB C ,Osteoblasts ,Tissue Engineering ,biology ,Cell Differentiation ,Mesenchymal Stem Cells ,Molecular biology ,In vitro ,medicine.anatomical_structure ,Osteocalcin ,biology.protein ,Alkaline phosphatase ,Bone marrow ,Biomedical engineering - Abstract
In this study we investigated not only the cellular proliferation and osteogenic differentiation of human bone marrow stromal cells (hBMSCs) on the novel beta-tricalcium phosphate (beta-TCP) scaffolds in vitro but also bone formation by ectopic implantation in athymic mice in vivo. The interconnected porous beta-TCP scaffolds with pores of 300-500 microm in size were prepared by the polymeric sponge method. beta-TCP scaffolds with the dimension of 3 mm x 3 mm x 3 mm were combined with hBMSCs, and incubated with (+) or without (-) osteogenic medium in vitro. Cell proliferation and osteogenic differentiation on the scaffolds were evaluated by scanning electron microscopy (SEM) observation, MTT assay, alkaline phosphatase (ALP) activity and osteocalcin (OCN) content measurement. SEM observation showed that hBMSCs attached well on the scaffolds and proliferated rapidly. No significant difference in the MTT assay could be detected between the two groups, but the ALP activity and OCN content of scaffolds (+) were much higher than those of the scaffolds (-) (p0.05). These results indicated that the novel porous beta-TCP scaffolds can support the proliferation and subsequent osteogenic differentiation of hBMSCs in vitro. After being cultured in vitro for 14 days, the scaffolds (+) and (-) were implanted into subcutaneous sites of athymic mice. In beta-TCP scaffolds (+), woven bone formed after 4 weeks of implantation and osteogenesis progressed with time. Furthermore, tissue-engineered bone could be found at 8 weeks, and remodeled lamellar bone was also observed at 12 weeks. However, no bone formation could be found in beta-TCP scaffolds (-) at each time point checked. The above findings illustrate that the novel porous beta-TCP scaffolds developed in this work have prominent osteoconductive activity and the potential for applications in bone tissue engineering.
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- 2007
6. Autologously generated tissue-engineered bone flaps for reconstruction of large mandibular defects in an ovine model
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Tang Ho, Jeroen J.J.P. van den Beucken, Patrick P. Spicer, Johnny Lam, F. Kurtis Kasper, John D. Jackson, John A. Jansen, James D. Kretlow, Yilin Cao, Guangpeng Liu, Mark E. Wong, Wei Liu, Antonios G. Mikos, Nagi Demian, Michael J. Miller, Alexander M. Tatara, James J. Yoo, Steven Lu, and Anthony Atala
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medicine.medical_specialty ,Scaffold ,Facial bone ,Materials science ,X-ray microtomography ,Biomedical Engineering ,Bioengineering ,Mandible ,Biochemistry ,Transplantation, Autologous ,Surgical Flaps ,Biomaterials ,Prosthesis Implantation ,Tissue engineering ,medicine ,Animals ,Polymethyl Methacrylate ,Craniofacial ,Sheep ,Tissue Engineering ,Original Articles ,X-Ray Microtomography ,Plastic Surgery Procedures ,Surgery ,Transplantation ,Disease Models, Animal ,Reconstructive and regenerative medicine Radboud Institute for Molecular Life Sciences [Radboudumc 10] ,Female ,Biomedical engineering - Abstract
Item does not contain fulltext The reconstruction of large craniofacial defects remains a significant clinical challenge. The complex geometry of facial bone and the lack of suitable donor tissue often hinders successful repair. One strategy to address both of these difficulties is the development of an in vivo bioreactor, where a tissue flap of suitable geometry can be orthotopically grown within the same patient requiring reconstruction. Our group has previously designed such an approach using tissue chambers filled with morcellized bone autograft as a scaffold to autologously generate tissue with a predefined geometry. However, this approach still required donor tissue for filling the tissue chamber. With the recent advances in biodegradable synthetic bone graft materials, it may be possible to minimize this donor tissue by replacing it with synthetic ceramic particles. In addition, these flaps have not previously been transferred to a mandibular defect. In this study, we demonstrate the feasibility of transferring an autologously generated tissue-engineered vascularized bone flap to a mandibular defect in an ovine model, using either morcellized autograft or synthetic bone graft as scaffold material. 01 mei 2015
- Published
- 2015
7. Age-Related Changes in the Regenerative Potential of Adipose-Derived Stem Cells Isolated from the Prominent Fat Pads in Human Lower Eyelids
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Xinhai Ye, Guangpeng Liu, Jian Tan, Zhenshun Song, Caihe Liao, and Yipin Xu
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0301 basic medicine ,Aging ,Pathology ,Cellular differentiation ,lcsh:Medicine ,Adipose tissue ,Alizarin Staining ,Cell Separation ,Biochemistry ,Regenerative medicine ,Fats ,0302 clinical medicine ,Tissue engineering ,Osteogenesis ,Animal Cells ,Adipocytes ,Medicine and Health Sciences ,Group-Specific Staining ,lcsh:Science ,Cellular Senescence ,Connective Tissue Cells ,Skin ,Staining ,Multidisciplinary ,Stem Cells ,Stem Cell Therapy ,Age Factors ,Cell Staining ,Cell Differentiation ,Middle Aged ,Flow Cytometry ,Lipids ,Adipose Tissue ,Connective Tissue ,Adipogenesis ,030220 oncology & carcinogenesis ,Female ,Cellular Types ,Anatomy ,Integumentary System ,Stem cell ,Chondrogenesis ,Orbit ,Cell aging ,Research Article ,Adult ,medicine.medical_specialty ,Biology ,Real-Time Polymerase Chain Reaction ,Research and Analysis Methods ,Young Adult ,03 medical and health sciences ,Internal medicine ,medicine ,Humans ,Regeneration ,Progenitor cell ,Cell Shape ,Aged ,Cell Proliferation ,Clinical Genetics ,lcsh:R ,Eyelids ,Biology and Life Sciences ,Cell Biology ,Kinetics ,Biological Tissue ,030104 developmental biology ,Endocrinology ,Specimen Preparation and Treatment ,lcsh:Q ,Developmental Biology - Abstract
The existence of multipotent adipose-derived stem cells isolated from human orbital fat (OF) tissue has shown great therapeutic potential in tissue engineering and regenerative medicine. But the use of stem cells for therapeutic applications is influenced by their proliferative and differentiation potentials, which may be affected by the age of the donor. So far there is little knowledge about the effects of donor age on the biological properties of human orbital adipose-derived stem cells (OASCs). The intraorbital fat protrusion in the lower eyelids occurs as an aging process, and the protruded fat is routinely removed during aesthetic surgeries. Based on the ease of OF harvest and the availability of OASCs, we investigated in this study the relationship between age and the differentiation and proliferation potentials of human OASCs. Human orbital adipose samples were harvested from young (with normal lower eyelid appearance) and old donors (having protruded fat pads in the lower eyelids). The morphological properties of orbital adipocytes were assessed and the fat cell size displayed a decreasing trend with advancing age. OASCs were isolated from the fat samples, expanded in vitro and cultured under appropriate inducive conditions. Compared to the young cells, although no difference was found in the cell yield and phenotype expression, aged OASCs showed fewer progenitor cell numbers, reduced proliferative rates, increased senescent features and decreased differentiation potentials towards adipogenic, osteogenic and chondrogenic lineages. Our data suggested that using autologous OASCs from elderly patients for potential therapeutic purposes might be restricted.
- Published
- 2016
8. Bone regeneration in a canine cranial model using allogeneic adipose derived stem cells and coral scaffold
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Lei Cui, Bo Liu, Yun Zhang, Guangpeng Liu, Jian Sun, and Wuyin Li
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Male ,medicine.medical_specialty ,Pathology ,Bone Regeneration ,animal diseases ,Biophysics ,Adipose tissue ,Bioengineering ,Bone healing ,Biology ,Bone and Bones ,Biomaterials ,Immune system ,Dogs ,In vivo ,Osteogenesis ,medicine ,Animals ,Transplantation, Homologous ,Bone regeneration ,Cells, Cultured ,Tissue Engineering ,Tissue Scaffolds ,Regeneration (biology) ,Stem Cells ,Cell Differentiation ,Mixed lymphocyte reaction ,Anthozoa ,Surgery ,Transplantation ,Adipose Tissue ,Mechanics of Materials ,Models, Animal ,Ceramics and Composites ,Female - Abstract
Adipose tissue derived stem cells (ASCs) based therapies for the repair and regeneration of various tissues have been widely investigated recently because of their multilineage potential and self-renewal capability. Our previous study demonstrated that autologous ASCs loaded onto natural coral scaffolds could repair cranial critical-sized defects (CSDs) in a canine model. The objective of this study was to determine whether the use of allogeneic ASCs could heal the same defect without the use of immunosuppressive therapy. The pedigree mismatch, mixed lymphocyte reaction assays (MLRs) and allogeneic skin graft experiments were performed to confirm unrelated ASC donors and recipients. A total of 12 adult Beagle dogs were enrolled in this study and divided into two groups. Bilateral cranial CSDs were created in each animal. The right-side defect was treated with allogeneic ASCs delivered onto a coral scaffold, and the left defect was either filled with an autologous ASC/coral composite (Group 1, n = 5) or with one coral scaffold alone (Group 2, n = 5). The systematic immune response and bone healing were evaluated postoperatively. The results showed that allogeneic ASC transplantation did not induce a systemic immune response by the hosts, and allogeneic ASCs could repair the cranial CSDs in an analogous way to that of the autologous cells. Moreover, both the green fluorescently labeled allogeneic and autologous ASCs were detected within the lacunae of newly formed bone in the defect site at 24 weeks, illustrating that the grafted ASCs contributed directly to bone regeneration in vivo. Thus, we concluded that allogeneic ASCs have the capacity to regenerate bone within craniofacial defects, providing an alternative source of seed cells for bone tissue engineering.
- Published
- 2012
9. Ectopic bone regeneration by human bone marrow mononucleated cells, undifferentiated and osteogenically differentiated bone marrow mesenchymal stem cells in beta-tricalcium phosphate scaffolds
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Jian Tan, Da-Wei Yang, Xinhai Ye, Guangpeng Liu, and Xiao-Fan Yin
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Adult ,Calcium Phosphates ,Male ,Pathology ,medicine.medical_specialty ,Bone Regeneration ,Adolescent ,Biomedical Engineering ,Medicine (miscellaneous) ,Mice, Nude ,Bioengineering ,Biocompatible Materials ,Bone Marrow Cells ,Bone healing ,Bone and Bones ,Mice ,Young Adult ,stomatognathic system ,Tissue engineering ,In vivo ,Osteogenesis ,Bone cell ,medicine ,Animals ,Humans ,Cells, Cultured ,Aged ,Cell Proliferation ,Mice, Inbred BALB C ,Tissue Engineering ,Tissue Scaffolds ,Chemistry ,Regeneration (biology) ,Cell Differentiation ,Mesenchymal Stem Cells ,Middle Aged ,In vitro ,Transplantation ,medicine.anatomical_structure ,Female ,Bone marrow ,Biomedical engineering - Abstract
Tissue engineering approaches using the combination of porous ceramics and bone marrow mesenchymal stem cells (BMSCs) represent a promising bone substitute for repairing large bone defects. Nevertheless, optimal conditions for constructing tissue-engineered bone have yet to be determined. It remains unclear if transplantation of predifferentiated BMSCs is superior to undifferentiated BMSCs or freshly isolated bone marrow mononucleated cells (BMNCs) in terms of new bone formation in vivo. The aim of this study was to investigate the effect of in vitro osteogenic differentiation (β-glycerophosphate, dexamethasone, and l-ascorbic acid) of human BMSCs on the capability to form tissue-engineered bone in unloaded conditions after subcutaneous implantation in nude mice. After isolation from human bone marrow aspirates, BMNCs were divided into three parts: one part was seeded onto porous beta-tricalcium phosphate ceramics immediately and transplanted in a heterotopic nude mice model; two parts were expanded in vitro to passage 2 before cell seeding and in vivo transplantation, either under osteogenic conditions or not. Animals were sacrificed for micro-CT and histological evaluation at 4, 8, 12, 16, and 20 weeks postimplantation. The results showed that BMSCs differentiated into osteo-progenitor cells after induction, as evidenced by the altered cell morphology and elevated alkaline phosphatase activity and calcium deposition, but their clonogenicity, proliferating rate, and seeding efficacy were not significantly affected by osteogenic differentiation, compared with undifferentiated cells. Extensive new bone formed in the pores of all the scaffolds seeded with predifferentiated BMSCs at 4 weeks after implantation, and maintained for 20 weeks. On the contrary, scaffolds containing undifferentiated BMSCs revealed limited bone formation only in 1 out of 6 cases at 8 weeks, and maintained for 4 weeks. For scaffolds with BMNCs, woven bone was observed sporadically only in one case at 8 weeks. Overall, this study suggests that ectopic osteogenesis of cell/scaffold composites is more dependent on the in vitro expansion condition, and osteo-differentiated BMSCs hold the highest potential concerning in vivo bone regeneration.
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- 2012
10. [Effect of cryopreservation on growth and osteogenesis of human adipose-derived stem cells]
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Guangpeng, Liu, Yulin, Li, Jian, Sun, Heng, Zhou, and Lei, Cui
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Cryopreservation ,Tissue Engineering ,Osteogenesis ,Stem Cells ,Adipocytes ,Humans ,Cell Differentiation ,Cells, Cultured - Abstract
As one of the adult stem cells, adipose-derived stem cells (ADSCs) have become an important seed cell source for tissue engineering recently. But whether the thawed cryopreserved ADSCs could be used to tissue engineered bone remains unknown. To investigate the effect of cryopreservation on the growth and osteogenesis of ADSCs in vitro.The ADSCs were isolated from the adipose aspirates by collagenase digestion method. For the experimental group, the 2nd generation cells were stored with a simple method of cryopreservation by slow cooling with dimethyl sulphoxide as a cryoprotectant and rapid thawing. After cryopreserved in liquid nitrogen for 4 weeks, ADSCs were recovered and cultured in osteogenic media, with non-cryopreserved ADSCs as the control group. The osteogenic differentiation was evaluated by alkaline phosphatase (ALP) staining and Alizarin red O staining at 2 and 3 weeks respectively. The cell growth and osteogenesis of ADSCs were further determined using DNA assay and the ALP activity and calcium content were measured.The survival percentage of the cryopreserved cells was 90.44% +/- 2.62%. The cell numbers and ALP activity increased with osteogenic induction time, and reach plateaus at 7 days and 11 days, respectively. The ALP staining and Alizarin red O staining results were both positive at 2 weeks and 3 weeks after osteogenic induction, respectively. And no significant difference in the cells number, ALP activity, and calcium content were found between experimental group and control group (P0.05).Cryopreservation does not affect the growth and osteogenesis of ADSCs, and the cryopreserved ADSCs can be used as cell source for tissue engineered bone.
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- 2010
11. Osteogenic differentiation of GFP-labeled human umbilical cord blood derived mesenchymal stem cells after cryopreservation
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Yulin Li, Yilin Cao, Jian Sun, Xinhai Ye, Guangpeng Liu, Lei Cui, and Yuchang Zhu
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Cellular differentiation ,Cell ,Green Fluorescent Proteins ,Transfection ,Umbilical cord ,General Biochemistry, Genetics and Molecular Biology ,Cryopreservation ,Green fluorescent protein ,fluids and secretions ,Tissue engineering ,In vivo ,Osteogenesis ,medicine ,Humans ,Cells, Cultured ,Chemistry ,Mesenchymal stem cell ,Cell Differentiation ,Mesenchymal Stem Cells ,General Medicine ,Fetal Blood ,Cell biology ,medicine.anatomical_structure ,Microscopy, Fluorescence ,embryonic structures ,Immunology ,General Agricultural and Biological Sciences - Abstract
The osteogenic capacity of human umbilical cord blood derived mesenchymal stem cells (UCB-MSCs) has been demonstrated both in vitro and in vivo. Therefore, cell labeling and storage are becoming necessary for researching the potential therapeutic use of UCB-MSCs for bone tissue engineering. The aim of this study was to determine the effect of cryopreservation on the osteogenic differentiation of green fluorescent protein (GFP)-marked UCB-MSCs in vitro. MSCs were isolated from full-term human UCB, expanded, transfected with the GFP gene, and then cryopreserved in liquid nitrogen for 4 weeks. After thawing, cell surface antigen markers and osteogenic potential were analyzed, and the luminescence of these cells was observed by fluorescence microscopy. The results demonstrate that cryopreservation has no effect on the cell phenotype, GFP expression or osteogenic differentiation of UCB-MSCs, showing that cryopreserved GFP-labeled UCB-MSCs might be applied for bone tissue engineering.
- Published
- 2010
12. Comparative investigation of viability, metabolism and osteogenic capability of tissue-engineered bone preserved in sealed osteogenic media at 37 °C and 4 °C
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Guangpeng Liu, Hengjian Wang, Yilin Cao, Lei Cui, Guangdong Zhou, and Lian Cen
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Adult ,Male ,Stromal cell ,Materials science ,Cell Survival ,Organ Preservation Solutions ,Biomedical Engineering ,Human bone ,Bioengineering ,Cryopreservation ,Biomaterials ,Osteogenesis ,medicine ,Humans ,Cells, Cultured ,Cell Proliferation ,Tissue Survival ,Ice formation ,Osteoblasts ,Tissue Engineering ,Tissue Scaffolds ,Demineralized bone matrix ,Cell Differentiation ,Mesenchymal Stem Cells ,Metabolism ,Cell biology ,medicine.anatomical_structure ,Tissue engineered bone ,Bone marrow ,Biomedical engineering - Abstract
Preservation of tissue-engineered (TE) bone is one of the key problems needed to be solved for its clinic application and industrialization. Traditional cryopreservation has been restricted because of the damages caused by ice formation and solution. Hypothermic preservation at 4 °C has been widely used for the preservation of transplanted organ despite potential negative effects on viability of cells and tissue. 37 °C is the best temperature for maintaining cellular bioactivities. However, 37 °C also has a potential negative effect on preserved cells due to consumption of nutrients and accumulation of by-products. No studies have reported which temperature is more suitable for the preservation of TE bone constructs. The current study explored the feasibility of preservation of TE bone constructs in sealed osteogenic media at 37 °C and 4 °C. Human bone marrow stromal cells (hBMSCs) were seeded into partially demineralized bone matrix (pDBM) scaffolds and cultured for 7 days to form TE bone constructs. The constructs were preserved in sealed osteogenic media at either 37 °C or 4 °C for 5, 7, 9 and 11 days, respectively. Growth kinetics, viability, metabolism and osteogenic capability were evaluated to explore the feasibility of preservation at 37 °C and 4 °C. The constructs cultured in osteogenic media at humidified 37 °C/5%CO2 served as the positive control. The results demonstrated that all the constructs preserved at 4 °C showed negative osteogenic capability at all time points with a much lower level of growth kinetics, viability and metabolism compared to the positive control. However, the constructs preserved at 37 °C showed good osteogenic capability within 7 days with a certain level of growth kinetics, viability and metabolism, although an obvious decrease in osteogenic capability was observed in the constructs preserved at 37 °C over 9 days. These results indicate that the preservation of TE bone constructs is feasible at 37 °C within 7 days in sealed osteogenic media.
- Published
- 2010
13. Repair of canine mandibular bone defects with bone marrow stromal cells and coral
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Guangpeng Liu, Zuo Liang Qi, Min Wei, Wenjie Zhang, Yi Lin Cao, Wei Liu, Jie Yuan, and Lei Cui
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Experimental control ,Stromal cell ,X-ray microtomography ,Coral ,Green Fluorescent Proteins ,Biomedical Engineering ,Bioengineering ,Bone Marrow Cells ,Biochemistry ,Biomaterials ,Dogs ,stomatognathic system ,Tissue engineering ,Osteogenesis ,Absorbable Implants ,Medicine ,Animals ,Bone formation ,Bone Marrow Transplantation ,Osteoblasts ,Tissue Engineering ,business.industry ,technology, industry, and agriculture ,Cell Differentiation ,Anatomy ,X-Ray Microtomography ,Anthozoa ,Mandibular Injuries ,Recombinant Proteins ,Biomechanical Phenomena ,medicine.anatomical_structure ,Bone Substitutes ,Bone marrow ,Stromal Cells ,business ,Immunostaining - Abstract
Tissue engineering has become a new approach for repairing bone defects. Previous studies indicated that coral scaffolds had been utilized with bone marrow stromal cells (BMSCs) in a variety of approaches for bony reconstruction. In these applications, the degradation rate of the material did not match the rate at which bone was regenerated. In this study, a previously established 30 mm long mandibular segmental defect was repaired with engineered bone using green fluorescent protein-labeled osteogenic BMSCs seeded on porous coral (n = 12). Defects treated with coral alone (n = 12) were used as an experimental control. In the BMSCs/coral group, new bone formation was observed from 4 weeks postoperation, and bony-union was achieved after 32 postoperative weeks. The residual coral volume of the BMSCs/coral grafts at 12 weeks (20-30%) was significantly higher than that at 32 weeks (10-15%, p < 0.05), which was detected by microcomputed tomography and histological examination. The engineered bone with BMSCs/coral achieved satisfactory biomechanical properties at 32 weeks postoperation, which was very close to that of the contralateral edentulous mandible. More importantly, immunostaining demonstrated that the implanted BMSCs differentiated into osteoblast-like cells. In contrast, minimal bone formation with almost solely fibrous connection was observed in the group treated with coral alone. Based on these results, we conclude that engineered bone from osteogenically induced BMSCs and biodegradable coral can successfully repair the critical-sized segmental mandibular defects in canines and the seeding cells could be used for bony restoration.
- Published
- 2009
14. The role of the extracellular signal-related kinase signaling pathway in osteogenic differentiation of human adipose-derived stem cells and in adipogenic transition initiated by dexamethasone
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Lei Cui, Qihai Liu, Wei Liu, Lian Cen, Yilin Cao, Guangpeng Liu, Heng Zhou, and Shuo Yin
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MAPK/ERK pathway ,Adult ,MAP Kinase Signaling System ,Biomedical Engineering ,Bioengineering ,Biology ,Biochemistry ,Dexamethasone ,Biomaterials ,Western blot ,Osteogenesis ,Extracellular ,medicine ,Adipocytes ,Humans ,Extracellular Signal-Regulated MAP Kinases ,Cells, Cultured ,Cell Proliferation ,Adipogenesis ,Osteoblasts ,medicine.diagnostic_test ,Tissue Engineering ,Kinase ,Cell Differentiation ,Cell biology ,Alkaline phosphatase ,Female ,Signal transduction ,Stem cell - Abstract
Human adipose-derived stem cells (hASCs) offer great promise for bone tissue engineering because of their osteogenic differentiation potential. At molecular levels, this study investigated the contribution of one of the main members of mitogen-activated protein kinases (MAPKs), extracellular signal-related kinase (ERK), to hASC osteogenic differentiation and the regulation of ERK for the balance between osteogenesis and adipogenesis in hASCs in vitro. As analyzed using western blot, ERK activation in osteo-induced hASCs was initiated at day 7, peaked at day 10, and declined from day 14 to basal levels. As detected using histochemical and biochemical methods, alkaline phosphatase (ALP) activity in hASCs experienced a process similar to that of ERK activation. Inhibition of ERK activation by PD98059, a specific inhibitor of the ERK signaling pathway, blocked the osteogenic differentiation in a dose-dependent manner, as revealed by an ALP activity assay, extracellular calcium deposition detection, osteocalcin (OCN) secretion examination, and real-time polymerase chain reaction (PCR) analysis for expression of osteogenesis-relative genes: core binding factor alpha 1, collagen type I, ALP, and OCN. Blockage of ERK phosphorylation in osteo-induced hASCs by PD98059 supplemented with dexamethasone (Dex) led to adipogenic differentiation, as confirmed by Nile Red staining to detect intracellular lipid droplets and real-time PCR analysis for expression of adipogenesis-relative genes: peroxisome proliferator-activated receptor gamma 2 and fatty acid-binding protein. These findings indicated a potential mechanism for the function of ERK in hASC osteogenic differentiation, especially the regulation of ERK in association with Dex for the balance between osteogenesis and adipogenesis, pointing out the significance of ERK signaling pathway for ASCs as a promising cell source for bone tissue engineering.
- Published
- 2009
15. In vitro engineering of fibrocartilage using CDMP1 induced dermal fibroblasts and polyglycolide
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Yilin Cao, Lei Cui, Guangpeng Liu, Lian Cen, Jian Sun, Heng Zhou, Guiqing Zhao, Wei Liu, and Shuo Yin
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Materials science ,Polyglycolide ,Polyesters ,Cell ,Biophysics ,Bioengineering ,Collagen Type I ,Biomaterials ,Glycosaminoglycan ,chemistry.chemical_compound ,Dogs ,Tissue engineering ,Growth Differentiation Factor 5 ,Gene expression ,medicine ,Animals ,Aggrecans ,Cell Shape ,Collagen Type II ,Cells, Cultured ,Cell Proliferation ,Glycosaminoglycans ,Tissue Engineering ,Tissue Scaffolds ,Gene Expression Profiling ,Fibrocartilage ,SOX9 Transcription Factor ,Dermis ,Fibroblasts ,Alkaline Phosphatase ,Molecular biology ,In vitro ,Staining ,Extracellular Matrix ,Collagen Type I, alpha 1 Chain ,medicine.anatomical_structure ,chemistry ,Mechanics of Materials ,Ceramics and Composites ,Polyglycolic Acid ,Biomedical engineering - Abstract
This study was designed to explore the feasibility of using cartilage-derived morphogenetic protein-1 (CDMP1) induced dermal fibroblasts (DFs) as seed cells and polyglycolide (PGA) as scaffold for fibrocartilage engineering. DFs isolated from canine were expanded and seeded on PGA scaffold to fabricate cell/scaffold constructs which were cultured with or without CDMP1. Proliferation and differentiation of DFs in different constructs were determined by DNA assay and glycosaminoglycan (GAG) production. Histological and immunohistochemical staining of the constructs after being in vitro cultured for 4 and 6 weeks were carried out to observe the fibrocartilage formation condition. The fibrocartilage-specific gene expression by cells in the constructs was analyzed by real-time PCR. It was shown that in the presence of CDMP1 the proliferation and GAG synthesis of DFs were significantly enhanced compared to those without CDMP1. Fibrocartilage-like tissue was formed in the CDMP1 induced construct after being cultured for 4 weeks, and it became more matured at 6 weeks as stronger staining for GAG and higher gene expression of collagen type II was observed. Since only weak staining for GAG and collagen type II was observed for the construct engineered without CDMP1, the induction effect on the fibrocartilage engineering can be ascertained when using DFs as seed cells. Furthermore, the potential of using DFs as seed cells to engineer fibrocartilage is substantiated and further study on using the engineered tissue to repair fibrocartilage defects is currently ongoing in our group.
- Published
- 2008
16. Evaluation of partially demineralized osteoporotic cancellous bone matrix combined with human bone marrow stromal cells for tissue engineering: an in vitro and in vivo study
- Author
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Jian Sun, Lei Cui, Guangpeng Liu, Yilin Cao, Wei Liu, Yulin Li, and Heng Zhou
- Subjects
Stromal cell ,Endocrinology, Diabetes and Metabolism ,Osteoporosis ,Cell Culture Techniques ,Dentistry ,Bone Matrix ,Bone Marrow Cells ,Bone healing ,Bone and Bones ,Mice ,Endocrinology ,Tissue engineering ,Implants, Experimental ,Osteogenesis ,Bone cell ,medicine ,Animals ,Humans ,Orthopedics and Sports Medicine ,Aged ,Aged, 80 and over ,Bone Transplantation ,Tissue Engineering ,Tissue Scaffolds ,Chemistry ,Demineralized bone matrix ,business.industry ,dBm ,Mesenchymal Stem Cells ,Middle Aged ,medicine.disease ,medicine.anatomical_structure ,Microscopy, Electron, Scanning ,Female ,Stromal Cells ,business ,Cancellous bone ,Biomedical engineering - Abstract
Allogenous demineralized bone matrix (DBM) represents a potential scaffold for bone tissue engineering due to its close relation in structure and function with autologous bone, but its supply is often restricted by donor availability. Thus, an expanded source of human bone is needed. The aim of this study was to evaluate the capacity of partially DBM scaffolds derived from allogenous cancellous bone of osteoporotic femurs to support osteogenesis of human bone marrow stromal cells (BMSCs) in vitro and in vivo in order to assess their potential use in bone tissue-engineering strategies. Human BMSCs of passage 2 were seeded either on osteoporotic bone-derived DBM scaffolds or on normal bone-derived scaffolds and cultured in osteogenic medium for 14 days. To assess the in vitro proliferation potential and osteogenic differentiation of BMSCs on scaffolds, scanning electronic microscopy observation, DNA content assays, and measurements of alkaline phosphatase activity and osteocalcin content were applied; the results displayed no significant differences between the osteoporotic DBM group and the normal DBM group. After 2 weeks of subculture in vitro, the BMSC/DBM composites were subcutaneously implanted into athymic mice for 8 weeks to evaluate their in vivo bone-forming ability. Histological examination showed tissue-engineered bone formation in the DBM pores in both groups, and no significant differences were observed in either the extent or frequency of new bone formation between these two groups. Based on these results, it can be concluded that osteoporotic bone-derived DBM may serve as a promising scaffold for bone tissue engineering.
- Published
- 2008
17. Repair of goat tibial defects with bone marrow stromal cells and beta-tricalcium phosphate
- Author
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Guangpeng Liu, Wenjie Zhang, Lei Cui, Li Zhao, Yilin Cao, and Wei Liu
- Subjects
Calcium Phosphates ,Materials science ,Stromal cell ,Bone density ,Polymers ,Biomedical Engineering ,Biophysics ,Bioengineering ,Biocompatible Materials ,Bone Marrow Cells ,Biomaterials ,Tissue engineering ,Bone Density ,medicine ,Animals ,Tibia ,Bone regeneration ,Bone mineral ,β tricalcium phosphate ,Tissue Engineering ,Goats ,Biomechanical Phenomena ,Radiography ,medicine.anatomical_structure ,Bone Substitutes ,Bone marrow ,Stromal Cells ,Tomography, X-Ray Computed ,Porosity ,Biomedical engineering - Abstract
Tissue engineering techniques have been proven effective in bone regeneration and repairing load-bearing bone defects. Previous studies, however, have heretofore been limited to the use of slowdegradable or natural biomaterials as scaffolds. There are, however, no reports on using biodegradable, synthetic beta-tricalcium phosphate (beta-TCP) as scaffolds to repair weight-bearing bone defects in large animals. In the present study, highly porous beta-TCP scaffolds prepared by the polymeric sponge method were used to repair goat tibial defects. Fifteen goats were randomly assigned to one of three groups, and a 26 mm-long defect at the middle part of the right tibia in each goat was created. In Group A (six goats), a porous beta-TCP ceramic cylinder that had been loaded with osteogenically induced autologous bone marrow stromal cells (BMSCs) was implanted in the defect of each animal. In Group B (six goats), the same beta-TCP ceramic cylinder without any cells loaded was placed in the defect. In Group C (three goats), the defect was left untreated. In Group A, bony union can be observed by gross view, X-ray and micro-computed tomography (Micro-CT) detection, and histological observation at 32 weeks post-implantation. The implanted beta-TCP scaffolds were almost completely replaced by tissue-engineered bone. Bone mineral density in the repaired area of Group A was significantly higher (p0.05) than that of Group B, in which scant new bone was formed in each defect and the beta-TCP hadn't been completely resorbed at 32 weeks. Moreover, the tissue-engineered bone of Group A had similar biomechanical properties as that of the normal left tibia in terms of bending strength and Young's modulus (p0.05). In Group C, little or no new bone was formed, and non-union occurred, showing that the 26 mm segmental defect of the goat tibia was critical sized at 32 weeks. Thus, it can be concluded that the mechanical properties of the BMSCs/beta-TCP composites could be much improved via tissue engineering approach and beta-TCP might be used to repair the weight-bearing segmental defects of goat tibias.
- Published
- 2007
18. Age-Related Changes in the Regenerative Potential of Adipose-Derived Stem Cells Isolated from the Prominent Fat Pads in Human Lower Eyelids.
- Author
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Ye, Xinhai, Liao, Caihe, Liu, Guangpeng, Xu, Yipin, Tan, Jian, and Song, Zhenshun
- Subjects
EYELID diseases ,REGENERATION (Biology) ,ADIPOSE tissues ,CELL differentiation ,CELL proliferation ,STEM cells ,TISSUE engineering ,PHYSIOLOGY - Abstract
The existence of multipotent adipose-derived stem cells isolated from human orbital fat (OF) tissue has shown great therapeutic potential in tissue engineering and regenerative medicine. But the use of stem cells for therapeutic applications is influenced by their proliferative and differentiation potentials, which may be affected by the age of the donor. So far there is little knowledge about the effects of donor age on the biological properties of human orbital adipose-derived stem cells (OASCs). The intraorbital fat protrusion in the lower eyelids occurs as an aging process, and the protruded fat is routinely removed during aesthetic surgeries. Based on the ease of OF harvest and the availability of OASCs, we investigated in this study the relationship between age and the differentiation and proliferation potentials of human OASCs. Human orbital adipose samples were harvested from young (with normal lower eyelid appearance) and old donors (having protruded fat pads in the lower eyelids). The morphological properties of orbital adipocytes were assessed and the fat cell size displayed a decreasing trend with advancing age. OASCs were isolated from the fat samples, expanded in vitro and cultured under appropriate inducive conditions. Compared to the young cells, although no difference was found in the cell yield and phenotype expression, aged OASCs showed fewer progenitor cell numbers, reduced proliferative rates, increased senescent features and decreased differentiation potentials towards adipogenic, osteogenic and chondrogenic lineages. Our data suggested that using autologous OASCs from elderly patients for potential therapeutic purposes might be restricted. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
19. The effect of fiber size and pore size on cell proliferation and infiltration in PLLA scaffolds on bone tissue engineering.
- Author
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Wang, Xuejun, Lou, Tao, Zhao, Wenhua, Song, Guojun, Li, Chunyao, and Cui, Guangpeng
- Subjects
PORE size (Materials) ,TISSUE scaffolds ,CELL proliferation ,TISSUE engineering ,CELL physiology ,LEACHING - Abstract
The scaffold microstructure has a great impact on cell functions in tissue engineering. Herein, the PLLA scaffolds with hierarchical fiber size and pore size were successfully fabricated by thermal-induced phase separation or combined thermal-induced phase separation and salt leaching methods. The PLLA scaffolds were fabricated as microfibrous scaffolds, microfibrous scaffolds with macropores (50–350 µm), nanofibrous scaffolds with micropores (100 nm to 10 µm), and nanofibrous scaffolds with both macropores and micropores by tailoring selective solvents for forming different fiber size and pre-sieved salts for creating controlled pore size. Among the four kinds of PLLA scaffolds, the nanofibrous scaffolds with both macropores and micropores provided a favorable microenvironment for protein adsorption, cell proliferation, and cell infiltration. The results further confirmed the significance of fiber size and pore size on the biological properties, and a scaffold with both micropores and macropores, and a nanofibrous matrix might have promising applications in bone tissue engineering. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
20. Bone marrow stromal cells with a combined expression of BMP-2 and VEGF-165 enhanced bone regeneration
- Author
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Yao Fu, Huifang Zhou, Hongliang Hou, Guangpeng Liu, Tingting Tang, Xianqun Fan, Caiwen Xiao, Ping Gu, and Peng Zhang
- Subjects
Vascular Endothelial Growth Factor A ,Bone Regeneration ,Stromal cell ,Materials science ,Biomedical Engineering ,Bone Morphogenetic Protein 2 ,Neovascularization, Physiologic ,Bone Marrow Cells ,Bioengineering ,Bone healing ,Bone morphogenetic protein 2 ,Bone remodeling ,Biomaterials ,Osteogenesis ,Transduction, Genetic ,Bone cell ,medicine ,Animals ,Humans ,Bone regeneration ,Orbital Fractures ,Cells, Cultured ,Cell Proliferation ,Tissue Engineering ,Tissue Scaffolds ,Cell Differentiation ,X-Ray Microtomography ,Anthozoa ,Recombinant Proteins ,Bone morphogenetic protein 7 ,medicine.anatomical_structure ,Bone Substitutes ,Cancer research ,Female ,Rabbits ,Bone marrow ,Stromal Cells ,Biomedical engineering - Abstract
Bone graft substitutes with osteogenic factors alone often exhibit poor bone regeneration due to inadequate vascularization. Combined delivery of osteogenic and angiogenic factors from biodegradable scaffolds may enhance bone regeneration. We evaluated the effects of bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor (VEGF), combined with natural coral scaffolds, on the repair of critical-sized bone defects in rabbit orbits. In vitro expanded rabbit bone marrow stromal cells (BMSCs) were transfected with human BMP2 and VEGF165 genes. Target protein expression and osteogenic differentiation were confirmed after gene transduction. Rabbit orbital defects were treated with a coral scaffold loaded with BMP2-transduced and VEGF-transduced BMSCs, BMP2-expressing BMSCs, VEGF-expressing BMSCs, or BMSCs without gene transduction. Volume and density of regenerated bone were determined by micro-computed tomography at 4, 8, and 16 weeks after implantation. Neovascularity, new bone deposition rate, and new bone formation were measured by immunostaining, tetracycline and calcein labelling, and histomorphometric analysis at different time points. The results showed that VEGF increased blood vessel formation relative to groups without VEGF. Combined delivery of BMP2 and VEGF increased new bone deposition and formation, compared with any single factor. These findings indicate that mimicking the natural bone development process by combined BMP2 and VEGF delivery improves healing of critical-sized orbital defects in rabbits.
- Published
- 2011
21. Bone regeneration in a canine cranial model using allogeneic adipose derived stem cells and coral scaffold
- Author
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Liu, Guangpeng, Zhang, Yun, Liu, Bo, Sun, Jian, Li, Wuyin, and Cui, Lei
- Subjects
- *
BONE regeneration , *ADIPOSE tissues , *STEM cells , *BIOLOGICAL assay , *TISSUE scaffolds , *TISSUE engineering , *AUTOPOIESIS , *MATHEMATICAL models - Abstract
Abstract: Adipose tissue derived stem cells (ASCs) based therapies for the repair and regeneration of various tissues have been widely investigated recently because of their multilineage potential and self-renewal capability. Our previous study demonstrated that autologous ASCs loaded onto natural coral scaffolds could repair cranial critical-sized defects (CSDs) in a canine model. The objective of this study was to determine whether the use of allogeneic ASCs could heal the same defect without the use of immunosuppressive therapy. The pedigree mismatch, mixed lymphocyte reaction assays (MLRs) and allogeneic skin graft experiments were performed to confirm unrelated ASC donors and recipients. A total of 12 adult Beagle dogs were enrolled in this study and divided into two groups. Bilateral cranial CSDs were created in each animal. The right-side defect was treated with allogeneic ASCs delivered onto a coral scaffold, and the left defect was either filled with an autologous ASC/coral composite (Group 1, n = 5) or with one coral scaffold alone (Group 2, n = 5). The systematic immune response and bone healing were evaluated postoperatively. The results showed that allogeneic ASC transplantation did not induce a systemic immune response by the hosts, and allogeneic ASCs could repair the cranial CSDs in an analogous way to that of the autologous cells. Moreover, both the green fluorescently labeled allogeneic and autologous ASCs were detected within the lacunae of newly formed bone in the defect site at 24 weeks, illustrating that the grafted ASCs contributed directly to bone regeneration in vivo. Thus, we concluded that allogeneic ASCs have the capacity to regenerate bone within craniofacial defects, providing an alternative source of seed cells for bone tissue engineering. [Copyright &y& Elsevier]
- Published
- 2013
- Full Text
- View/download PDF
22. Osteogenic differentiation of GFP-labeled human umbilical cord blood derived mesenchymal stem cells after cryopreservation
- Author
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Liu, Guangpeng, Ye, Xinhai, Zhu, Yuchang, Li, Yulin, Sun, Jian, Cui, Lei, and Cao, Yilin
- Subjects
- *
CORD blood , *MESENCHYMAL stem cells , *CRYOPRESERVATION of organs, tissues, etc. , *TISSUE engineering , *GREEN fluorescent protein , *CELL differentiation , *FLUORESCENCE microscopy , *BONE growth - Abstract
Abstract: The osteogenic capacity of human umbilical cord blood derived mesenchymal stem cells (UCB-MSCs) has been demonstrated both in vitro and in vivo. Therefore, cell labeling and storage are becoming necessary for researching the potential therapeutic use of UCB-MSCs for bone tissue engineering. The aim of this study was to determine the effect of cryopreservation on the osteogenic differentiation of green fluorescent protein (GFP)-marked UCB-MSCs in vitro. MSCs were isolated from full-term human UCB, expanded, transfected with the GFP gene, and then cryopreserved in liquid nitrogen for 4weeks. After thawing, cell surface antigen markers and osteogenic potential were analyzed, and the luminescence of these cells was observed by fluorescence microscopy. The results demonstrate that cryopreservation has no effect on the cell phenotype, GFP expression or osteogenic differentiation of UCB-MSCs, showing that cryopreserved GFP-labeled UCB-MSCs might be applied for bone tissue engineering. [Copyright &y& Elsevier]
- Published
- 2011
- Full Text
- View/download PDF
23. Evaluation of Partially Demineralized Osteoporotic Cancellous Bone Matrix Combined with Human Bone Marrow Stromal Cells for Tissue Engineering: An In Vitro and In Vivo Study.
- Author
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Guangpeng Liu, Jian Sun, Yulin Li, Heng Zhou, Lei Cui, Wei Liu, and Yilin Cao
- Subjects
- *
BONES , *FEMUR , *BONE marrow , *ALKALINE phosphatase , *TISSUE engineering , *OSTEOPOROSIS - Abstract
Allogenous demineralized bone matrix (DBM) represents a potential scaffold for bone tissue engineering due to its close relation in structure and function with autologous bone, but its supply is often restricted by donor availability. Thus, an expanded source of human bone is needed. The aim of this study was to evaluate the capacity of partially DBM scaffolds derived from allogenous cancellous bone of osteoporotic femurs to support osteogenesis of human bone marrow stromal cells (BMSCs) in vitro and in vivo in order to assess their potential use in bone tissue-engineering strategies. Human BMSCs of passage 2 were seeded either on osteoporotic bone–derived DBM scaffolds or on normal bone–derived scaffolds and cultured in osteogenic medium for 14 days. To assess the in vitro proliferation potential and osteogenic differentiation of BMSCs on scaffolds, scanning electronic microscopy observation, DNA content assays, and measurements of alkaline phosphatase activity and osteocalcin content were applied; the results displayed no significant differences between the osteoporotic DBM group and the normal DBM group. After 2 weeks of subculture in vitro, the BMSC/DBM composites were subcutaneously implanted into athymic mice for 8 weeks to evaluate their in vivo bone-forming ability. Histological examination showed tissue-engineered bone formation in the DBM pores in both groups, and no significant differences were observed in either the extent or frequency of new bone formation between these two groups. Based on these results, it can be concluded that osteoporotic bone–derived DBM may serve as a promising scaffold for bone tissue engineering. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
24. Evaluation of the viability and osteogenic differentiation of cryopreserved human adipose-derived stem cells
- Author
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Liu, Guangpeng, Zhou, Heng, Li, Yulin, Li, Gang, Cui, Lei, Liu, Wei, and Cao, Yilin
- Subjects
- *
CRYOPRESERVATION of organs, tissues, etc. , *STEM cells , *ADIPOSE tissues , *BONE cells - Abstract
Abstract: Human adipose-derived stem cells (ASCs) have the ability to differentiate into osteoblasts and thus the potential therapeutic use to tissue-engineer bone, so a reliable method for cell storage is necessary. The aim of this study was to determine whether a simple method of cryopreservation with 10% Me2SO as a protectant had an effect on proliferation potential and osteogenic differentiation of ASCs isolated from fresh human adipose tissue. ASCs were harvested from 6 human lipoaspirates and each was halved for either cryopreservation in liquid nitrogen for 2 weeks or for control culture. Cells from the second-passage were plated at a density of 5000cells/well in 24-well plates and cultured with or without osteogenic media for 14 days. Cell surface antigens were used to identify the cryopreserved ASCs by flow cytometry. The proliferation rate of both populations was evaluated using a cell DNA assay. To detect osteogenic differentiation of both the cryopreserved and non-cryopreserved populations, determination of osteoblastic protein production (alkaline phosphatase and osteocalcin) and excellular matrix calcification (calcium content) was applied. The expression of osteoblastic-associated genes was also analyzed using reverse-transcription polymerase chain reaction. These results demonstrate that cryopreservation has no effect on the phenotype, proliferation or osteogenic differentiation of human ASCs, showing cryopreserved human ASCs might be applied for bone tissue engineering. [Copyright &y& Elsevier]
- Published
- 2008
- Full Text
- View/download PDF
25. Tissue-engineered bone formation with cryopreserved human bone marrow mesenchymal stem cells
- Author
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Liu, Guangpeng, Shu, Chaofeng, Cui, Lei, Liu, Wei, and Cao, Yilin
- Subjects
- *
CRYOPRESERVATION of organs, tissues, etc. , *IMMUNE system , *ALKALINE phosphatase , *LIQUID nitrogen - Abstract
Abstract: Bone marrow mesenchymal stem cells (MSCs) have become the main cell source for bone tissue engineering. It has been reported that cryopreserved human MSCs can maintain their potential for proliferation and osteogenic differentiation in vitro. There are, however, no reports on osteogenesis with cryopreserved human MSCs in vivo. The aim of this study was to determine whether cryopreservation had an effect on the proliferation capability and osteogenic differentiation of human MSCs on scaffolds in vitro and in vivo. MSCs were isolated from human bone marrow, cultured in vitro until passage 2, and then frozen and stored at −196°C in liquid nitrogen with 10% Me2SO as cryoprotectant for 24h. The cryopreserved MSCs were then thawed rapidly, seeded onto partially demineralized bone matrix (pDBM) scaffolds and cultured in osteogenic media containing 10mM sodium β-glycerophosphate, 50μM l-ascorbic acid, and 10nM dexamethasone. Non-cryopreserved MSCs seeded onto the pDBM scaffolds were used as control groups. Scanning electronic microscopy (SEM) observation, DNA content assays, and measurements of alkaline phosphatase (ALP) activity and osteocalcin (OCN) content were applied, and the results showed that the proliferation potential and osteogenic differentiation of MSCs on pDBM in vitro were not affected by cryopreservation. After 2 weeks of subculture, the MSCs/pDBM composites were subcutaneously implanted into the athymic mice. The constructs were harvested at 4 and 8 weeks postimplantation, and histological examination showed tissue-engineered bone formation in the pDBM pores in both groups. Based on these results, it can be concluded that cryopreservation allows human MSCs to be available for potential therapeutic use to tissue-engineer bone. [Copyright &y& Elsevier]
- Published
- 2008
- Full Text
- View/download PDF
26. Repair of goat tibial defects with bone marrow stromal cells and β-tricalcium phosphate.
- Author
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Liu, Guangpeng, Zhao, Li, Zhang, Wenjie, Cui, Lei, Liu, Wei, and Cao, Yilin
- Subjects
GOATS ,TIBIA ,BONE marrow ,CALCIUM phosphate ,TISSUE engineering ,BONE regeneration ,CELLS ,BONES ,LEG - Abstract
Tissue engineering techniques have been proven effective in bone regeneration and repairing load-bearing bone defects. Previous studies, however, have heretofore been limited to the use of slowdegradable or natural biomaterials as scaffolds. There are, however, no reports on using biodegradable, synthetic beta-tricalcium phosphate (β-TCP) as scaffolds to repair weight-bearing bone defects in large animals. In the present study, highly porous β-TCP scaffolds prepared by the polymeric sponge method were used to repair goat tibial defects. Fifteen goats were randomly assigned to one of three groups, and a 26 mm-long defect at the middle part of the right tibia in each goat was created. In Group A (six goats), a porous β-TCP ceramic cylinder that had been loaded with osteogenically induced autologous bone marrow stromal cells (BMSCs) was implanted in the defect of each animal. In Group B (six goats), the same β-TCP ceramic cylinder without any cells loaded was placed in the defect. In Group C (three goats), the defect was left untreated. In Group A, bony union can be observed by gross view, X-ray and micro-computed tomography (Micro-CT) detection, and histological observation at 32 weeks post-implantation. The implanted β-TCP scaffolds were almost completely replaced by tissue-engineered bone. Bone mineral density in the repaired area of Group A was significantly higher ( p < 0.05) than that of Group B, in which scant new bone was formed in each defect and the β-TCP hadn’t been completely resorbed at 32 weeks. Moreover, the tissue-engineered bone of Group A had similar biomechanical properties as that of the normal left tibia in terms of bending strength and Young’s modulus ( p > 0.05). In Group C, little or no new bone was formed, and non-union occurred, showing that the 26 mm segmental defect of the goat tibia was critical sized at 32 weeks. Thus, it can be concluded that the mechanical properties of the BMSCs/β-TCP composites could be much improved via tissue engineering approach and β-TCP might be used to repair the weight-bearing segmental defects of goat tibias. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
27. Vitreous cryopreservation of tissue engineered bone composed of bone marrow mesenchymal stem cells and partially demineralized bone matrix
- Author
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Yin, Hongyu, Cui, Lei, Liu, Guangpeng, Cen, Lian, and Cao, Yilin
- Subjects
- *
CRYOPRESERVATION of organs, tissues, etc. , *BONE marrow , *STEM cells , *MESENCHYME , *TISSUE engineering , *MOLECULAR structure , *CELL proliferation , *CRYOBIOLOGY ,PRESERVATION - Abstract
Abstract: Cryopreservation of tissue engineered products by maintaining their structure and function is a prerequisite for large-scale clinical applications. In this study, we examined the feasibility of cryopreservation of tissue engineered bone (TEB) composed of osteo-induced canine bone marrow mesenchymal stem cells (cBMSCs) and partially demineralized bone matrix (pDBM) scaffold by vitrification. A novel vitreous solution named as VS442 containing 40% dimethyl-sulfoxide (DMSO), 40% EuroCollins (EC) solution and 20% basic culture medium (BCM) was developed. After being cultured in vitro for 8days, cell/scaffold complex in VS442 was subjected to vitreous preservation for 7days and 3months, respectively. Cell viability, proliferation and osteogenic differentiation of cBMSCs in TEB after vitreous cryopreservation were examined with parallel comparisons being made with those cryopreserved in VS55 vitreous solution. Compared with that cryopreserved in VS55, cell viability and subsequent proliferative ability of TEB in VS442 after being rewarmed were significantly higher as detected by live/dead staining and DNA assay. The level of alkaline phosphatase (ALP) expression and osteocalcin (OCN) deposition in VS442 preserved TEB was also higher than those in the VS55 group since 3days post-rewarm. Both cell viability and osteogenic capability of the VS55 group were found to be declined to a negligible level within 15days post-rewarm. Furthermore, it was observed that extending the preservation of TEB in VS442 to 3months did not render any significant effect on its survival and osteogenic potential. Thus, the newly developed VS442 vitreous solution was demonstrated to be more efficient in maintaining cellular viability and osteogenic function for vitreous cryopreservation of TEB over VS55. [Copyright &y& Elsevier]
- Published
- 2009
- Full Text
- View/download PDF
28. In vitro engineering of fibrocartilage using CDMP1 induced dermal fibroblasts and polyglycolide
- Author
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Zhao, Guiqing, Yin, Shuo, Liu, Guangpeng, Cen, Lian, Sun, Jian, Zhou, Heng, Liu, Wei, Cui, Lei, and Cao, Yilin
- Subjects
- *
TISSUE engineering , *FIBROBLASTS , *CARTILAGE , *FEASIBILITY studies , *BIOMEDICAL materials , *BONE morphogenetic proteins , *CELL proliferation , *IMMUNOHISTOCHEMISTRY - Abstract
Abstract: This study was designed to explore the feasibility of using cartilage-derived morphogenetic protein-1 (CDMP1) induced dermal fibroblasts (DFs) as seed cells and polyglycolide (PGA) as scaffold for fibrocartilage engineering. DFs isolated from canine were expanded and seeded on PGA scaffold to fabricate cell/scaffold constructs which were cultured with or without CDMP1. Proliferation and differentiation of DFs in different constructs were determined by DNA assay and glycosaminoglycan (GAG) production. Histological and immunohistochemical staining of the constructs after being in vitro cultured for 4 and 6 weeks were carried out to observe the fibrocartilage formation condition. The fibrocartilage-specific gene expression by cells in the constructs was analyzed by real-time PCR. It was shown that in the presence of CDMP1 the proliferation and GAG synthesis of DFs were significantly enhanced compared to those without CDMP1. Fibrocartilage-like tissue was formed in the CDMP1 induced construct after being cultured for 4 weeks, and it became more matured at 6 weeks as stronger staining for GAG and higher gene expression of collagen type II was observed. Since only weak staining for GAG and collagen type II was observed for the construct engineered without CDMP1, the induction effect on the fibrocartilage engineering can be ascertained when using DFs as seed cells. Furthermore, the potential of using DFs as seed cells to engineer fibrocartilage is substantiated and further study on using the engineered tissue to repair fibrocartilage defects is currently ongoing in our group. [Copyright &y& Elsevier]
- Published
- 2009
- Full Text
- View/download PDF
29. In vitro generation of an osteochondral construct using injectable hydrogel composites encapsulating rabbit marrow mesenchymal stem cells
- Author
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Guo, Xuan, Park, Hansoo, Liu, Guangpeng, Liu, Wei, Cao, Yilin, Tabata, Yasuhiko, Kasper, F. Kurtis, and Mikos, Antonios G.
- Subjects
- *
HYDROGELS , *COMPOSITE materials , *MICROENCAPSULATION , *MESENCHYME , *BONE marrow cells , *STEM cells , *TISSUE engineering , *LABORATORY rabbits - Abstract
Abstract: Injectable, biodegradable hydrogel composites of crosslinked oligo(poly(ethylene glycol) fumarate) (OPF) and gelatin microparticles (MPs) were utilized to fabricate a bilayered osteochondral construct consisting of a chondrogenic layer and an osteogenic layer, and to investigate the differentiation of rabbit marrow mesenchymal stem cells (MSCs) encapsulated in both layers in vitro. The results showed that MSCs in the chondrogenic layer were able to undergo chondrogenic differentiation, especially in the presence of TGF-β1-loaded MPs. In the osteogenic layer, cells maintained their osteoblastic phenotype. Although calcium deposition in the osteogenic layer was limited, cells in the osteogenic layer significantly enhanced chondrogenic differentiation of MSCs in the chondrogenic layer. The greatest effect was observed when MSCs were encapsulated with TGF-β1-loaded MPs and cultured with osteogenic cells in the bilayered constructs. Overall, this study demonstrates the fabrication of bilayered hydrogel composites that mimic the structure and function of osteochondral tissue, along with the application of these composites as cell and growth factor carriers. [Copyright &y& Elsevier]
- Published
- 2009
- Full Text
- View/download PDF
30. Adipose-derived stem cells alleviate osteoporosis by enchancing osteogenesis and inhibiting adipogenesis in a rabbit model.
- Author
-
XINHAI YE, PENG ZHANG, SHAOBO XUE, YIPIN XU, JIAN TAN, and GUANGPENG LIU
- Subjects
- *
OSTEOPOROSIS treatment , *STEM cells , *BONE growth , *BONE regeneration , *TISSUE engineering , *OVARIECTOMY , *ALGINIC acid - Abstract
Background aims. Osteoporosis (OP) is characterized by a reduction in bone quality, which is associated with inadequacies in bone marrow mesenchymal stromal cells (BMSCs). As an alternative cell source to BMSCs, adipose-derived stem cells (ASCs) have been investigated for bone repair because of their osteogenic potential and self-renewal capability. Nevertheless, whether autologous ASCs can be used to promote bone regeneration under osteoporotic conditions has not been elucidated. Methods. The OP rabbit model was established by means of bilateral ovariectomy (OVX). Both BMSCs and ASCs were harvested from OVX rabbits and expanded in vitro. The effects of osteogenic-induced ASCs on the in vitro adipogenic and osteogenic capabilities of BMSCs were evaluated. Autologous ASCs were then encapsulated by calcium alginate gel and transplanted into the distal femurs of OVX rabbits (n = 12). Hydrogel without loading cells was injected into the contralateral femurs as a control. Animals were killed for investigation at 12 weeks after transplantation. Results. Osteogenic-induced ASCs were able to promote osteogenesis and inhibit adipogenesis of osteoporotic BMSCs through activation of the bone morphogenetic protein 2/ bone morphogenetic protein receptor type IB signal pathway. Local bone mineral density began to increase at 8 weeks after ASC transplantation (P < 0.05). At 12 weeks, micro--computed tomography and histological evaluation revealed more new bone formation in the cell-treated femurs than in the control group (P < 0.05). Conclusions. This study demonstrated that ASCs could stimulate proliferation and osteogenic differentiation of BMSCs in vitro and enhance bone regeneration in vivo, which suggests that autologous osteogenic-induced ASCs might be useful to alleviate OP temporally. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
31. Repair of articular cartilage defect in non-weight bearing areas using adipose derived stem cells loaded polyglycolic acid mesh
- Author
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Cui, Lei, Wu, Yaohao, Cen, Lian, Zhou, Heng, Yin, Shuo, Liu, Guangpeng, Liu, Wei, and Cao, Yilin
- Subjects
- *
ARTICULAR cartilage , *WOUND healing , *FAT cells , *STEM cells , *BIODEGRADABLE plastics , *CHONDROGENESIS , *CELL differentiation , *TISSUE engineering - Abstract
Abstract: The current study was designed to observe chondrogenic differentiation of adipose derived stem cells (ASCs) on fibrous polyglycolic acid (PGA) scaffold stabilized with polylactic acid (PLA), and to further explore the feasibility of using the resulting cell/scaffold constructs to repair full thickness articular cartilage defects in non-weight bearing area in porcine model within a follow-up of 6 months. Autologous ASCs isolated from subcutaneous fat were expanded and seeded on the scaffold to fabricate ASCs/PGA constructs. Chondrogenic differentiation of ASCs in the constructs under chondrogenic induction was monitored with time by measuring the expression of collagen type II (COL II) and glycosaminoglycan (GAG). The constructs after being in vitro induced for 2 weeks were implanted to repair full thickness articular cartilage defects (8mm in diameter, deep to subchondral bone) in femur trochlea (the experimental group), while scaffold alone was implanted to serve as the control. Histologically, the generated neo-cartilage integrated well with its surrounding normal cartilage and subchondral bone in the defects of experimental group at 3 months post-implantation, whereas only fibrous tissue was filled in the defects of control group. Immunohistochemical and toluidine blue staining confirmed the similar distribution of COL II and GAG in the regenerated cartilage as the normal one. A vivid remolding process with post-operation time was also witnessed in the neo-cartilage as its compressive moduli increased significantly from 50.55% of the normal cartilage at 3 months to 88.05% at 6 months. The successful repair thus substantiates the potentiality of using chondrogenic induced ASCs and PGA/PLA scaffold for cartilage regeneration. [Copyright &y& Elsevier]
- Published
- 2009
- Full Text
- View/download PDF
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