Effect of Strontium Enhanced Calcium Phosphate Coating on In Vitro Behavior of Human Mesenchymal Stem Cell (hMSC)

Author(s):  
Samuel C. Uzoechi ◽  
Goddy C. Okoye ◽  
Kennedy O. Ejeta ◽  
Benjamin I. Nkem ◽  
Gideon I. Ndubuka

Calcium phosphate is a widely used material as coating for metallic implants. This research describes a biomimetic coating techniques based on deposition of calcium phosphate films on a Ti6Al4V plates that was used to study the effect of strontium additive on the behavior of hMSCs. In this study, strontium additive was homogenously deposited onto calcium phosphate films on a Ti6AlV plates by using a biomimetic techniques. Strontium affected composition and morphology of calcium phosphate deposited on a Ti6Al4V plates to a varying degree, according to concentration of solutions used. The effect of strontium additive on proliferation and differentiation of hMSCs depended on the solution and concentration tested. In general, all individual three coatings showed decreased hMSCs proliferation. Strontium additive demonstrated a significant increase in differentiation into osteogenic lineage when compared with the control and calcium phosphate films without strontium additive. However, no cytotoxic effect of strontium additive in the concentrations tested was detected. The Fourier transform infrared spectra showed that this new coating closely resembles bone mineral. The techniques illustrated in this study mimics bone mineral containing strontium additive, making it constructive for studying basic processes of in vitro bone formation. The results showed in this study can be used for changing bone graft substitutes by addition of strontium additive on implants in order to affect their performance in bone repair and regeneration. Also, the system can aid rapid bone formation around the implant, reducing therewith the patient’s recovery time after surgery.

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Xiaohua Yu ◽  
Mei Wei

The influence of biomimetic calcium phosphate coating on osteoblasts behaviorin vitrois not well established yet. In this study, we investigated the behavior of osteoblastic rat osteosarcoma 17/2.8 cells (ROS17/2.8) on two groups of biomaterial surfaces: alkaline-treated titanium surface (ATT) and biomimetic calcium phosphate coated ATT (CaP). The cell attachment, proliferation, differentiation, and morphology on these surfaces were extensively evaluated to reveal the impact of substrate surface on osteoblastic cell responses. It was found that the ROS17/2.8 cells cultured on the ATT surface had higher attachment and proliferation rates compared to those on the CaP surface. Our results also showed that the calcium phosphate coatings generated in this work have an inhibiting effect on osteoblast adhesion and further influenced the proliferation and differentiation of osteoblast compared to the ATT surfacein vitro. Cells on the ATT surface also exhibited a higher alkaline phosphatase activity than on the CaP surface after two weeks of culture. Immunofluorescence staining and scanning electron microscopy results showed that the cells adhered and spread faster on the ATT surface than on the CaP surface. These results collectively suggested that substrate surface properties directly influence cell adhesion on different biomaterials, which would result in further influence on the cell proliferation and differentiation.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Meadhbh Á. Brennan ◽  
Mario Barilani ◽  
Francesco Rusconi ◽  
Julien de Lima ◽  
Luciano Vidal ◽  
...  

AbstractBone marrow mesenchymal stem/stromal cells (BMSCs) show great promise for bone repair, however they are isolated by an invasive bone marrow harvest and their regenerative potential decreases with age. Conversely, cord blood can be collected non-invasively after birth and contains MSCs (CBMSCs) that can be stored for future use. However, whether CBMSCs can replace BMSCs targeting bone repair is unknown. This study evaluates the in vitro osteogenic potential of unprimed, osteogenically primed, or chondrogenically primed CBMSCs and BMSCs and their in vivo bone forming capacity following ectopic implantation on biphasic calcium phosphate ceramics in nude mice. In vitro, alkaline phosphatase (intracellular, extracellular, and gene expression), and secretion of osteogenic cytokines (osteoprotegerin and osteocalcin) was significantly higher in BMSCs compared with CBMSCs, while CBMSCs demonstrated superior chondrogenic differentiation and secretion of interleukins IL-6 and IL-8. BMSCs yielded significantly more cell engraftment and ectopic bone formation compared to CBMSCs. However, priming of CBMSCs with either chondrogenic or BMP-4 supplements led to bone formation by CBMSCs. This study is the first direct quantification of the bone forming abilities of BMSCs and CBMSCs in vivo and, while revealing the innate superiority of BMSCs for bone repair, it provides avenues to induce osteogenesis by CBMSCs.


Materials ◽  
2021 ◽  
Vol 14 (13) ◽  
pp. 3693
Author(s):  
Yurii P. Sharkeev ◽  
Ekaterina G. Komarova ◽  
Valentina V. Chebodaeva ◽  
Mariya B. Sedelnikova ◽  
Aleksandr M. Zakharenko ◽  
...  

A modern trend in traumatology, orthopedics, and implantology is the development of materials and coatings with an amorphous–crystalline structure that exhibits excellent biocopatibility. The structure and physico–chemical and biological properties of calcium phosphate (CaP) coatings deposited on Ti plates using the micro-arc oxidation (MAO) method under different voltages (200, 250, and 300 V) were studied. Amorphous, nanocrystalline, and microcrystalline statesof CaHPO4 and β-Ca2P2O7were observed in the coatings using TEM and XRD. The increase in MAO voltage resulted in augmentation of the surface roughness Ra from 2.5 to 6.5 µm, mass from 10 to 25 mg, thickness from 50 to 105 µm, and Ca/P ratio from 0.3 to 0.6. The electrical potential (EP) of the CaP coatings changed from −456 to −535 mV, while the zeta potential (ZP) decreased from −53 to −40 mV following an increase in the values of the MAO voltage. Numerous correlations of physical and chemical indices of CaP coatings were estimated. A decrease in the ZP magnitudes of CaP coatings deposited at 200–250 V was strongly associated with elevated hTERT expression in tumor-derived Jurkat T cells preliminarily activated with anti-CD2/CD3/CD28 antibodies and then contacted in vitro with CaP-coated samples for 14 days. In turn, in vitro survival of CD4+ subsets was enhanced, with proinflammatory cytokine secretion of activated Jurkat T cells. Thus, the applied MAO voltage allowed the regulation of the physicochemical properties of amorphous–crystalline CaP-coatings on Ti substrates to a certain extent. This method may be used as a technological mechanism to trigger the behavior of cells through contact with micro-arc CaP coatings. The possible role of negative ZP and Ca2+ as effectors of the biological effects of amorphous–crystalline CaP coatings is discussed. Micro-arc CaP coatings should be carefully tested to determine their suitability for use in patients with chronic lymphoid malignancies.


2007 ◽  
Vol 22 (6) ◽  
pp. 1593-1600 ◽  
Author(s):  
Sahil Jalota ◽  
Sutapa Bhaduri ◽  
Sarit B. Bhaduri ◽  
A. Cuneyt Tas

Biomimetic coating of titanium and related alloys with carbonated apatitic calcium phosphate is an important area of research in implantology. While this paper specifically refers to coating Ti6Al4V, the results are valid with other related alloys as well. One step in the protocol involves an intermediate alkali treatment of Ti6Al4V to form a sodium titanate layer on the alloy surface. This pretreatment enhances the formation of the coating from simulated body fluid (SBF) solutions. Many papers in the biomimetic coating literature demonstrate the presence of cracks in coatings, irrespective of the SBF compositions and placement of the substrates. The presence of cracks may result in degradation and delamination of coatings. To the best of our knowledge, this issue remains unresolved. Therefore, the aim of this study was: (i) to examine and understand the reasons for cracking and (ii) based on the results, to develop a protocol for producing crack-free apatitic calcium phosphate coatings on Ti6Al4V substrates. In this study, the authors focused their attention on the alkali treatment procedure and the final drying step. It is hypothesized that these two steps of the process affect the crack formation the most. In the first case, the surfaces of alkali-treated substrates were examined with/without water-soaking treatment before immersing in SBF. This water treatment modifies the sodium titanate surface layer. In the second case, two different drying techniques (after soaking in SBF) were used. In one procedure, the coated substrates were dried rapidly, and in the other they were dried slowly. It was observed that the water treatment, irrespective of the drying method, provides a surface, which on subsequent soaking in SBF forms a crack-free apatitic calcium phosphate coating. Based on these results, the authors suggest a protocol incorporating a water-soaking treatment after the alkali treatment and prior to the SBF soaking treatment to obtain crack-free coatings.


2009 ◽  
Vol 631-632 ◽  
pp. 211-216 ◽  
Author(s):  
Kyosuke Ueda ◽  
Takayuki Narushima ◽  
Takashi Goto ◽  
T. Katsube ◽  
Hironobu Nakagawa ◽  
...  

Calcium phosphate coating films were fabricated on Ti-6Al-4V plates and screw-type implants with a blast-treated surface using radiofrequency (RF) magnetron sputtering and were evaluated in vitro and in vivo. Amorphous calcium phosphate (ACP) and oxyapatite (OAp) films obtained in this study could cover the blast-treated substrate very efficiently, maintaining the surface roughness. For the in vitro evaluations of the calcium phosphate coating films, bonding strength and alkaline phosphatase (ALP) activity were examined. The bonding strength of the coating films to a blast-treated substrate exceeded 60 MPa, independent of film phases except for the film after post-heat-treatment in silica ampoule. When compared with an uncoated substrate, the increase in the ALP activity of osteoblastic SaOS-2 cells on a calcium phosphate coated substrate was confirmed by a cell culture test. The removal torque of screw-type Ti-6Al-4V implants with a blast-treated surface from the femur of Japanese white rabbit increased with the duration of implantation and it was statistically improved by coating an ACP film 2 weeks after implantation. The in vitro and in vivo studies suggested that the application of the sputtered ACP film as a coating on titanium implants was effective in improving their biocompatibility with bones.


2011 ◽  
Vol 5 (1) ◽  
pp. 63-71 ◽  
Author(s):  
Oliver D Schneider ◽  
Dirk Mohn ◽  
Roland Fuhrer ◽  
Karina Klein ◽  
Käthi Kämpf ◽  
...  

Background: The purpose of this preliminary study was to assess the in vivo performance of synthetic, cotton wool-like nanocomposites consisting of a biodegradable poly(lactide-co-glycolide) fibrous matrix and containing either calcium phosphate nanoparticles (PLGA/CaP 60:40) or silver doped CaP nanoparticles (PLGA/Ag-CaP 60:40). Besides its extraordinary in vitro bioactivity the latter biomaterial (0.4 wt% total silver concentration) provides additional antimicrobial properties for treating bone defects exposed to microorganisms. Materials and Methods: Both flexible artificial bone substitutes were implanted into totally 16 epiphyseal and metaphyseal drill hole defects of long bone in sheep and followed for 8 weeks. Histological and histomorphological analyses were conducted to evaluate the biocompatibility and bone formation applying a score system. The influence of silver on the in vivo performance was further investigated. Results: Semi-quantitative evaluation of histology sections showed for both implant materials an excellent biocompatibility and bone healing with no resorption in the adjacent bone. No signs of inflammation were detectable, either macroscopically or microscopically, as was evident in 5 µm plastic sections by the minimal amount of inflammatory cells. The fibrous biomaterials enabled bone formation directly in the centre of the former defect. The area fraction of new bone formation as determined histomorphometrically after 8 weeks implantation was very similar with 20.5 ± 11.2 % and 22.5 ± 9.2 % for PLGA/CaP and PLGA/Ag-CaP, respectively. Conclusions: The cotton wool-like bone substitute material is easily applicable, biocompatible and might be beneficial in minimal invasive surgery for treating bone defects.


2005 ◽  
Vol 288-289 ◽  
pp. 429-432 ◽  
Author(s):  
Zhi Qing Chen ◽  
Quan Li Li ◽  
Quan Zen ◽  
Gang Li ◽  
Hao Bin Jiang ◽  
...  

Phosphorylated chitosans were synthesized as templates to manipulate hydroxyapatite (HA) crystal nucleation, growth and microstructure. Two kinds of insoluble phosphorylated chitosan were soaked in saturated Ca(OH)2 solution for 4 d and in 1.5× SBF (simulated body fluid) solutions for 14 d at 37 °C for biomimetic mineralization. A lower [P]-content of phosphorylated chitosan promoted greater mineralization than higher [P]-content. Phosphorylated chitosan inhibited osteoblast proliferation and differentiation in vitro, while calcium phosphate phosphorylated chitosan composites did not.


2010 ◽  
Vol 654-656 ◽  
pp. 2065-2070
Author(s):  
Ho Yeon Song ◽  
Young Hee Kim ◽  
Jyoti M. Anirban ◽  
In Seon Byun ◽  
Kyung A Kwak ◽  
...  

Calcium phosphate ceramics such as hydroxy apatite (HA), β-tricalcium phosphate (β-TCP) and bicalcium phosphate (BCP) have been used as a bone graft biomaterial because of their good biocompatibility and similarity of chemical composition to natural bones. To increase the mechanical and osteoconductive properties, the granules and spongy type porous bone graft substitutes were prepared by fibrous monolithic process and polyurethane foam replica methods, respectively. The pore sizes obtained using these approaches ranged between 100-600 µm. The cytotoxicity, cellular proliferation, differentiation and ECM deposition on the bone graft substitutes were observed by SEM and confocal microscopy. Moreover, the scaffolds were implanted in the rabbit femur. New bone formation and biodegradation of bone graft were observed through follow-up X-ray, micro-CT analysis and histological findings. After several months (2, 3, 6, 12 and 24 months) of implantation, new bone formation and ingrowths were observed in defect sites of the animal by CaP ceramics and 2 to 3 times higher bone ingrowths were confirmed than that of the normal trabecular bones in terms of total bone volume (BV).


2016 ◽  
Vol 720 ◽  
pp. 90-94
Author(s):  
Masanobu Kamitakahara ◽  
Takashi Shirato ◽  
Taishi Yokoi ◽  
Hideaki Matsubara ◽  
Yasuaki Shibata ◽  
...  

Silicate-containing alpha-tricalcium phosphate (α-TCP) ceramics are expected to be useful scaffolds for bone regeneration because α-TCP shows high biodegradability and silicate ions are expected to promote the bone formation. We previously revealed that the porous silicate-containing α-TCP granules provided earlier bone formation and showed lower biodegradability than the porous silicate-free α-TCP granules in vivo. In order to reveal the mechanism of the bone formation promoted by silicate incorporation, the proliferation and differentiation of osteoblast-like cells on the silicate-containing and silicate-free α-TCP ceramics were examined in vitro. The silicate incorporation in α-TCP promoted the differentiation of osteoblast-like cells, and it might be one of the factors to promote bone formation In Vivo.


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