Study on Bone Cell Adaptability of α-TCP/HAp Functionally Graded Porous Beads for Biomaterials Application

2010 ◽  
Vol 76 ◽  
pp. 143-146
Author(s):  
S. Ohtake ◽  
T. Asaoka ◽  
K. Furukawa ◽  
T. Ushida ◽  
T. Tateishi
Keyword(s):  
Bone Cells ◽  
High Strength ◽  
Bone Cell ◽  
Functionally Graded ◽  
The Body ◽  
Tissue Adhesive ◽  
Adhesive Properties ◽  
Living Body ◽  
Porous Beads

Porous beads of bioactive ceramics such as HAp, TCP are considered to be promising as excellent scaffolds for cultivating bone cells. To realize this type of beads which maintains the function of scaffold with sufficient strength up to growth of new bone, and is expected to absorbed completely after the growth, a-TCP/ HAp functionally graded porous beads were fabricated. HAp is bioactive material which has both high strength and better tissue-adhesive properties, but that is not readily absorbed by the human body. On the contrary, a-TCP is highly bioabsorbable; it is quickly absorbed by the body, and, therefore, disappears before bone is completely replaced. Fabricated new beads are composed of a-TCP at the center and HAp at the surface, to control the solubility in living body. Bone cell adaptability of these beads were confirmed in vitro.

Solubility Control of αTCP-HAp Functionally Graded Porous Beads in SBF for Biomaterial Use

2010 ◽  
Vol 638-642 ◽  
pp. 2021-2027
Author(s):  
Teruo Asaoka ◽  
Yuji Kajihata ◽  
Katsuko Furukawa ◽  
Takashi Ushida ◽  
Tetsuya Tateishi
Keyword(s):  
Body Fluid ◽  
Absorption Rate ◽  
Bone Cells ◽  
High Strength ◽  
Functionally Graded ◽  
The Body ◽  
Tissue Adhesive ◽  
Adhesive Properties ◽  
Living Body ◽  
Porous Beads

As excellent scaffolds for cultivating bone cells, porous beads of bioactive ceramics such as HAp, TCP are considered to be promising. HAp and α-TCP are well known as non-toxic bioceramics to human cells, but their behavior in living body fluid are different. HAp is bioactive material which has both high strength and better tissue-adhesive properties, but that is not readily absorbed by the human body. On the contrary, α-TCP is highly bioabsorbable; it is quickly absorbed by the body, and, therefore, disappears before bone is completely replaced. To realize a composite which has suitable solubility in living body fluid, α-TCP/ HAp functionally graded porous beads were fabricated by the method of spheroidization in liquid nitrogen. This type of composite maintains the function of scaffold with sufficient strength up to growth of new bone, and after the growth, it is expected to absorbed completely in the body. In the present study, ceramic beads with α-TCP at the center were fabricated and coated with a functionally graded layer of HAp. By controlling the thickness of HAp layer, which could be realized by changing time of hydrolytic reaction, the absorption rate into the body would be easily controlled. In addition, to accelerate the formation of porous structure, some acid solutions were used to dissolve the beads surface and to penetrate pores toward inside of the beads. Observed surface and inner structure by SEM, also the measured change in absorption rate will be presented.

Fabrication of α-TCP, HAp Functionally Graded Porous Beads

2008 ◽  
Vol 57 ◽  
pp. 135-138
Author(s):  
Yuji Kajihata ◽  
Teruo Asaoka ◽  
Katsuko S. Furukawa ◽  
Takashi Ushida ◽  
T. Tateishi
Keyword(s):  
Surface Layer ◽  
High Strength ◽  
Functionally Graded ◽  
The Body ◽  
Acid Solutions ◽  
Bioactive Material ◽  
Hydrolytic Reaction ◽  
Rate Of Absorption ◽  
Porous Beads ◽  
Good Cell

HAp (Hydroxyapatite) and α-TCP (alpha tribasic calcium phosphate) are non-toxic to human cells and, thus, have been studied for applications as biomaterials. HAp is a bioactive material that is not readily absorbed by the body; it offers both high strength and better tissueadhesive properties than α-TCP. In contrast, α-TCP is highly bioabsorbable; it is quickly absorbed by the body, and, therefore, for example, disappears before bone is completely replaced. If porous beads could be fabricated that would take advantage of the useful properties of α-TCP and HAp, they could be used as excellent scaffolds for cultivating cells. In the present study, ceramic beads with α-TCP at the center were fabricated and coated with a functionally graded film of HAp. A scaffold based on this configuration would be expected to have the following characteristics: good cell adhesion; strong beads; and a rate of absorption into the body that would be easy to control. In addition, to accelerate the formation of porous structure, some acid solutions were used to dissolve the beads surface layer and to penetrate pores toward inside of the bead. HAp formation through hydrolytic reaction seemed to be promoted by these acid solutions.

scholarly journals Protocol of Co-Culture of Human Osteoblasts and Osteoclasts to Test Biomaterials for Bone Tissue Engineering

2022 ◽  
Vol 5 (1) ◽  
pp. 8
Author(s):  
Giorgia Borciani ◽  
Giorgia Montalbano ◽  
Nicola Baldini ◽  
Chiara Vitale-Brovarone ◽  
Gabriela Ciapetti
Keyword(s):  
Tissue Engineering ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Bone Cells ◽  
Bone Cell ◽  
Seeding Density ◽  
Bone Homeostasis ◽  
Bone Microenvironment

New biomaterials and scaffolds for bone tissue engineering (BTE) applications require to be tested in a bone microenvironment reliable model. On this assumption, the in vitro laboratory protocols with bone cells represent worthy experimental systems improving our knowledge about bone homeostasis, reducing the costs of experimentation. To this day, several models of the bone microenvironment are reported in the literature, but few delineate a protocol for testing new biomaterials using bone cells. Herein we propose a clear protocol to set up an indirect co-culture system of human-derived osteoblasts and osteoclast precursors, providing well-defined criteria such as the cell seeding density, cell:cell ratio, the culture medium, and the proofs of differentiation. The material to be tested may be easily introduced in the system and the cell response analyzed. The physical separation of osteoblasts and osteoclasts allows distinguishing the effects of the material onto the two cell types and to evaluate the correlation between material and cell behavior, cell morphology, and adhesion. The whole protocol requires about 4 to 6 weeks with an intermediate level of expertise. The system is an in vitro model of the bone remodeling system useful in testing innovative materials for bone regeneration, and potentially exploitable in different application fields. The use of human primary cells represents a close replica of the bone cell cooperation in vivo and may be employed as a feasible system to test materials and scaffolds for bone substitution and regeneration.

Rapid in situ cross-linking of hydrogel adhesives based on thiol-grafted bio-inspired catechol-conjugated chitosan

2017 ◽  
Vol 32 (5) ◽  
pp. 612-621 ◽  
Author(s):  
Zhiwen Zeng ◽  
Xiumei Mo
Keyword(s):  
Soft Tissues ◽  
Sodium Periodate ◽  
Oxidation Mechanism ◽  
Thiol Group ◽  
In Vitro Cytotoxicity ◽  
Tissue Adhesive ◽  
Molar Ratio ◽  
Adhesive Properties

In this paper, a novel chitosan derivative, thiol-grafting bio-inspired catechol-conjugated chitosan was synthesized. The chemical structure of the synthesized catechol-conjugated chitosan was verified by 1H NMR, and its contents of thiol group and catechol group were determined by UV-vis spectrum. Four percent of catechol-conjugated chitosan aqueous solution could form hydrogels rapidly in situ in 1 min or less with the addition of sodium periodate. Rheological studies showed that the mechanical properties depend on the concentrations of catechol-conjugated chitosan and the molar ratio of sodium periodate to catechol groups. Additionally, the adhesive properties of the resulting adhesives were evaluated, and the adhesion strength of obtained adhesives was as high as 50 kPa because of the complex and multiple interactions, especially the anti-oxidation mechanism of thiol group. The in vitro cytotoxicity assays demonstrated an excellent biocompatibility of the catechol-conjugated chitosan hydrogels. Benefiting from the in situ fast cured, desired mechanical strength, biocompatibility and relatively high adhesion performance, these properties suggested that catechol-conjugated chitosan hydrogel adhesives have potential applications as tissue adhesive for soft tissues.

Control of Oriented Extracellular Matrix Similar to Anisotropic Bone Microstructure

2014 ◽  
Vol 783-786 ◽  
pp. 72-77 ◽  
Author(s):  
Takayoshi Nakano ◽  
Aira Matsugaki ◽  
Takuya Ishimoto ◽  
Mitsuharu Todai ◽  
Ai Serizawa ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Bone Tissue ◽  
Bone Cells ◽  
Crystal Surface ◽  
Early Stage ◽  
Bone Microstructure ◽  
Bone Cell ◽  
Collagen Fibers

Bone microstructure is dominantly composed of anisotropic extracellular matrix (ECM) in which collagen fibers and epitaxially-oriented biological apatite (BAp) crystals are preferentially aligned depending on the bone anatomical position, resulting in exerting appropriate mechanical function. The regenerative bone in bony defects is however produced without the preferential alignment of collagen fibers and the c-axis of BAp crystals, and subsequently reproduced to recover toward intact alignment. Thus, it is necessary to produce the anisotropic bone-mimetic tissue for the quick recovery of original bone tissue and the related mechanical ability in the early stage of bone regeneration. Our group is focusing on the methodology for regulating the arrangement of bone cells, the following secretion of collagen and the self-assembled mineralization by oriented BAp crystallites. Cyclic stretching in vitro to bone cells, principal-stress loading in vivo on scaffolds, step formation by slip traces on Ti single crystal, surface modification by laser induced periodic surface structure (LIPSS), anisotropic collagen substrate with the different degree of orientation, etc. can dominate bone cell arrangement and lead to the construction of the oriented ECM similar to the bone tissue architecture. This suggests that stress/strain loading, surface topography and chemical anisotropy are useful to produce bone-like microstructure in order to promote the regeneration of anisotropic bone tissue and to understand the controlling parameters for anisotropic osteogenesis induction.

scholarly journals Natural and Synthetic Polymers for Bone Scaffolds Optimization

Polymers ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 905 ◽  
Author(s):  
Francesca Donnaloja ◽  
Emanuela Jacchetti ◽  
Monica Soncini ◽  
Manuela T. Raimondi
Keyword(s):  
Tissue Engineering ◽  
Bone Tissue ◽  
Bone Cells ◽  
The Body ◽  
Bone Tissue Regeneration ◽  
In Vivo Models ◽  
Polymeric Scaffold ◽  
Technology Improvement

Bone tissue is the structural component of the body, which allows locomotion, protects vital internal organs, and provides the maintenance of mineral homeostasis. Several bone-related pathologies generate critical-size bone defects that our organism is not able to heal spontaneously and require a therapeutic action. Conventional therapies span from pharmacological to interventional methodologies, all of them characterized by several drawbacks. To circumvent these effects, tissue engineering and regenerative medicine are innovative and promising approaches that exploit the capability of bone progenitors, especially mesenchymal stem cells, to differentiate into functional bone cells. So far, several materials have been tested in order to guarantee the specific requirements for bone tissue regeneration, ranging from the material biocompatibility to the ideal 3D bone-like architectural structure. In this review, we analyse the state-of-the-art of the most widespread polymeric scaffold materials and their application in in vitro and in vivo models, in order to evaluate their usability in the field of bone tissue engineering. Here, we will present several adopted strategies in scaffold production, from the different combination of materials, to chemical factor inclusion, embedding of cells, and manufacturing technology improvement.

Response of normal and osteoporotic human bone cells to mechanical stress in vitro

1998 ◽  
Vol 274 (6) ◽  
pp. E1113-E1120 ◽  
Author(s):  
Jozien G. H. Sterck ◽  
Jenneke Klein-Nulend ◽  
Paul Lips ◽  
Elisabeth H. Burger
Keyword(s):  
Nitric Oxide ◽  
Mechanical Stress ◽  
Cell Cultures ◽  
Bone Cells ◽  
Bone Cell ◽  
Human Bone ◽  
Animal Bone ◽  
Bone Cell Cultures

Bone adapts to mechanical stress, and bone cell cultures from animal origin have been shown to be highly sensitive to mechanical stress in vitro. In this study, we tested whether bone cell cultures from human bone biopsies respond to stress in a similar manner as animal bone cells and whether bone cells from osteoporotic patients respond similarly to nonosteoporotic donors. Bone cell cultures were obtained as outgrowth from collagenase-stripped trabecular bone fragments from 17 nonosteoporotic donors between 7 and 77 yr of age and from 6 osteoporotic donors between 42 and 72 yr of age. After passage, the cells were mechanically stressed by treatment with pulsating fluid flow (PFF; 0.7 ± 0.03 Pa at 5 Hz for 1 h) to mimic the stress-driven flow of interstitial fluid through the bone canaliculi, which is likely the stimulus for mechanosensation in bone in vivo. Similar to earlier studies in rodent and chicken bone cells, the bone cells from nonosteoporotic donors responded to PFF with enhanced release of prostaglandin E2(PGE2) and nitric oxide as well as a reduced release of transforming growth factor-β (TGF-β). The upregulation of PGE2 but not the other responses continued for 24 h after 1 h of PFF treatment. The bone cells from osteoporotic donors responded in a similar manner as the nonosteoporotic donors except for the long-term PGE2 release. The PFF-mediated upregulation of PGE2 release during 24 h of postincubation after 1 h of PFF was significantly reduced in osteoporotic patients compared with six age-matched controls as well as with the whole nonosteoporotic group. These results indicate that enhanced release of PGE2 and nitric oxide, as well as reduced release of TGF-β, is a characteristic response of human bone cells to fluid shear stress, similar to animal bone cells. The results also suggest that bone cells from osteoporotic patients may be impaired in their long-term response to mechanical stress.

Physiological Levels of Substrate Deformation Are Less Stimulatory to Bone Cells Compared to Fluid Flow

1999 ◽  
Author(s):  
Jun You ◽  
Clare E. Yellowley ◽  
Henry J. Donahue ◽  
Christopher R. Jacobs
Keyword(s):  
Fluid Flow ◽  
Matrix Protein ◽  
Bone Cells ◽  
Bone Matrix ◽  
Biological Response ◽  
Bone Cell ◽  
Osteoblastic Cells ◽  
Physiological Strain ◽  
Substrate Strain

Abstract It is believed that bone cells can sense mechanical loading and alter bone external shape and internal structure to efficiently support the load bearing demands placed upon it. However, the mechanism by which bone cells sense and respond to their mechanical environment is still poorly understood. In particular, the load-induced signals to which bone cells respond, e.g. fluid flow, substrate deformation, electrokinetic effects etc., are unclear. Furthermore, there are few studies focused on the effects of physiological strain (strain < 0.5%, Burr, 1996; Owan, 1997) on bone cells. The goal of this study was to investigate cytosolic Ca2+ mobilization (a very early signaling event) in response to different substrate strains (physiological or supra-physiological strains), and to distinguish the effects of substrate strain from those of fluid flow by applying precisely controlled strain without induced fluid flow. In addition, we quantified the effect of physiologically relevant fluid flow (Cowin, 1995) and substrate stretch on the expression of mRNA for the bone matrix protein osteopontin (OPN). A computer controlled stretch device was employed to apply different substrate strains, 0.1%, 1%, 5% and 10%. A parallel plate flow chamber was used to test cell responses to steady and oscillating flows (20dyn/cm2, 1Hz). Our data demonstrate that physiological strain (< 0.5%) does not induce [Ca2+]i responses in primary rat osteoblastic cells (ROB) in vitro. However, there was a significant (p < 0.05) increase in the number of responding cells at supra-physiological strains of 1, 5, and 10% suggesting that the cells were capable of a biological response. Similar results for human fetal osteoblastic cells (hFOB 1.19) and osteocyte-like cells (ML0-Y4) were obtained. Furthermore, compared to physiological substrate deformation, physiological fluid flow induced greater [Ca2+]i responses for hFOB cells, and these [Ca2+]i responses were quantitatively similar to those obtained for 10% substrate strain. Moreover we found no change in osteopontin mRNA expression after 0.5% strain stretch. Conversely, physiological oscillating flow (20dyn/cm2, 1Hz) caused a significant increase in osteopontin mRNA. These data suggest that, relative to fluid flow, substrate deformation may play less of a role in bone cell mechanotransduction.

scholarly journals Isolation of bone cell clones with differences in growth, hormone responses, and extracellular matrix production.

1982 ◽  
Vol 92 (2) ◽  
pp. 452-461 ◽  
Author(s):  
J E Aubin ◽  
J N Heersche ◽  
M J Merrilees ◽  
J Sodek
Keyword(s):  
Extracellular Matrix ◽  
Bone Cells ◽  
Bone Cell ◽  
Population Doubling ◽  
Cell Populations ◽  
Type I ◽  
Mixed Cell ◽  
Extracellular Matrix Components ◽  
Wide Range

Clones of nontransformed hormone-responsive bone cells have been isolated in vitro from mixed cell populations of fetal rat calvaria. In several independent isolations, microscopically visible colonies appeared at plating efficiencies of 5-10% of the starting cell numbers. Of these clones, approximately 10% grew to mass populations which could be assayed for a number of growth and biochemical properties. Although some similarities existed among the clones, they could be distinguished from each other and from the mixed cell populations. Population-doubling times (tDs) and saturation densities varied over a wide range: e.g., tDs of 24-72 h and saturation densities of 0.4-5 x 10(5) cells/cm2. Morphologies varied from roughly polygonal multilayering cells to typically spindle-shaped monolayering cells. Hormone responsiveness, as measured by stimulation of cAMP by hormones, indicated that some clones were responsive to both parathyroid hormone (PTH) and prostaglandin E2 (PGE2), while others responded to PTH only. Analysis of extracellular matrix components revealed that all clones produced type I and type III collagens, though in different proportions. Similarly, although all clones synthesized four glycosaminoglycans (hyaluronic acid, heparan sulfate, chondroitin sulfate, and dermatan sulfate), the quantities of each were distinctive from clone to clone. Further investigation of such clones is continuing to define more precisely the heterogeneity of clonal bone cell populations in vitro. They represent an important step in the study of the endocrinology and differentiation of bone.

scholarly journals Regenerative Medicine for Diabetes Mellitus

2009 ◽  
Vol 18 (5-6) ◽  
pp. 491-496 ◽  
Author(s):  
Naoya Kobayashi ◽  
Takeshi Yuasa ◽  
Teru Okitsu
Keyword(s):  
Regenerative Medicine ◽  
The Body ◽  
Definitive Treatment ◽  
Β Cells ◽  
Β Cell ◽  
Pancreatic Β Cells ◽  
Living Body

In diabetes, a loss of pancreatic β-cells causes insulin dependency. When insulin dependency is caused by type 1 diabetes or pancreatic diabetes, for example, pancreatic β-cells need to be regenerated for definitive treatment. The methods for generating pancreatic β-cells include a method of creating pancreatic β-cells in vitro and implanting them into the body and a method of regenerating pancreatic β-cells in the body via gene introduction or the administration of differential proliferation factors to the body. Moreover, the number of pancreatic β-cells is also low in type 2 diabetes, caused by the compounding factors of insulin secretory failure and insulin resistance; therefore, if pancreatic β-cells can be regenerated in a living body, then a further amelioration of the pathology can be expected. The development of pancreatic β-cell-targeting regenerative medicine can lead to the next generation of diabetes treatment.

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