Exploring the Biomaterial-Induced Secretome: Physical Bone Substitute Characteristics Influence the Cytokine Expression of Macrophages

In addition to their chemical composition various physical properties of synthetic bone substitute materials have been shown to influence their regenerative potential and to influence the expression of cytokines produced by monocytes, the key cell-type responsible for tissue reaction to biomaterials in vivo. In the present study both the regenerative potential and the inflammatory response to five bone substitute materials all based on β-tricalcium phosphate (β-TCP), but which differed in their physical characteristics (i.e., granule size, granule shape and porosity) were analyzed for their effects on monocyte cytokine expression. To determine the effects of the physical characteristics of the different materials, the proliferation of primary human osteoblasts growing on the materials was analyzed. To determine the immunogenic effects of the different materials on human peripheral blood monocytes, cells cultured on the materials were evaluated for the expression of 14 pro- and anti-inflammatory cytokines, i.e., IL-6, IL-10, IL-1β, VEGF, RANTES, IL-12p40, I-CAM, IL-4, V-CAM, TNF-α, GM-CSF, MIP-1α, Il-8 and MCP-1 using a Bio-Plex^® Multiplex System. The granular shape of bone substitutes showed a significant influence on the osteoblast proliferation. Moreover, smaller pore sizes, round granular shape and larger granule size increased the expression of GM-CSF, RANTES, IL-10 and IL-12 by monocytes, while polygonal shape and the larger pore sizes increased the expression of V-CAM. The physical characteristics of a bone biomaterial can influence the proliferation rate of osteoblasts and has an influence on the cytokine gene expression of monocytes in vitro. These results indicate that the physical structure of a biomaterial has a significant effect of how cells interact with the material. Thus, specific characteristics of a material may strongly affect the regenerative potential in vivo.     

In Vivo Analysis of the Biocompatibility and Bone Healing Capacity of a Novel Bone Grafting Material Combined with Hyaluronic Acid

The present in vivo study analyses both the inflammatory tissue reactions and the bone healing capacity of a newly developed bone substitute material (BSM) based on xenogeneic bone substitute granules combined with hyaluronate (HY) as a water-binding molecule. The results of the hyaluronate containing bone substitute material (BSM) were compared to a control xenogeneic BSM of the same chemical composition and a sham operation group up to 16 weeks post implantationem. A major focus of the study was to analyze the residual hyaluronate and its effects on the material-dependent healing behavior and the inflammatory tissue responses. The study included 63 male Wistar rats using the calvaria implantation model for 2, 8, and 16 weeks post implantationem. Established and Good Laboratory Practice (GLP)-conforming histological, histopathological, and histomorphometrical analysis methods were conducted. The results showed that the new hyaluronate containing BSM was gradually integrated within newly formed bone up to the end of the study that ended in a condition of complete bone defect healing. Thereby, no differences to the healing capacity of the control BSM were found. However, the bone formation in both groups was continuously significantly higher compared to the sham operation group. Additionally, no differences in the (inflammatory) tissue response that was analyzed via qualitative and (semi-) quantitative methods were found. Interestingly, no differences were found between the numbers of pro- and anti-inflammatory macrophages between the three study groups over the entire course of the study. No signs of the HY as a water-binding part of the BSM were histologically detectable at any of the study time points, altogether the results of the present study show that HY allows for an optimal material-associated bone tissue healing comparable to the control xenogeneic BSM. The added HY seems to be degraded within a very short time period of less than 2 weeks so that the remaining BSM granules allow for a gradual osteoconductive bone regeneration. Additionally, no differences between the inflammatory tissue reactions in both material groups and the sham operation group were found. Thus, the new hyaluronate containing xenogeneic BSM and also the control BSM have been shown to be fully biocompatible without any differences regarding bone regeneration.     

Synergistic Effect of PVDF-Coated PCL-TCP Scaffolds and Pulsed Electromagnetic Field on Osteogenesis

Bone exhibits piezoelectric properties. Thus, electrical stimulations such as pulsed electromagnetic fields (PEMFs) and stimuli-responsive piezoelectric properties of scaffolds have been investigated separately to evaluate their efficacy in supporting osteogenesis. However, current understanding of cells responding under the combined influence of PEMF and piezoelectric properties in scaffolds is still lacking. Therefore, in this study, we fabricated piezoelectric scaffolds by functionalization of polycaprolactone-tricalcium phosphate (PCL-TCP) films with a polyvinylidene fluoride (PVDF) coating that is self-polarized by a modified breath-figure technique. The osteoinductive properties of these PVDF-coated PCL-TCP films on MC3T3-E1 cells were studied under the stimulation of PEMF. Piezoelectric and ferroelectric characterization demonstrated that scaffolds with piezoelectric coefficient d₃₃ = −1.2 pC/N were obtained at a powder dissolution temperature of 100 °C and coating relative humidity (RH) of 56%. DNA quantification showed that cell proliferation was significantly enhanced by PEMF as low as 0.6 mT and 50 Hz. Hydroxyapatite staining showed that cell mineralization was significantly enhanced by incorporation of PVDF coating. Gene expression study showed that the combination of PEMF and PVDF coating promoted late osteogenic gene expression marker most significantly. Collectively, our results suggest that the synergistic effects of PEMF and piezoelectric scaffolds on osteogenesis provide a promising alternative strategy for electrically augmented osteoinduction. The piezoelectric response of PVDF by PEMF, which could provide mechanical strain, is particularly interesting as it could deliver local mechanical stimulation to osteogenic cells using PEMF.     

A new application of Hydroxyapatite extracted from dromedary bone: Adsorptive removal of Congo red from aqueous solution

The aim of this paper is to evaluate the efficiency of the natural hydroxyapatite powder for anionic dye (Congo red) elimination from water and optimization of adsorption process parameters that are contact time (0-360 min), solution pH (4-9), initial dye concentration (400-2400 mg/l), and adsorbent dosage (0.4-20 g/l). Natural hydroxyapatite with high surface area was prepared from animal agriculture waste dromedary bone. X-ray diffraction, transmission electron microscopy, transform infrared spectroscopy, and elemental analysis characterizations of the powder prepared show a hexagonal structure, the irregular form of hydroxyapatite and presence of other elements as trace element. The results of adsorption tests demonstrated the high efficiency of natural hydroxyapatite for removal of Congo red from water solution whose maximum experimental capacity adsorption of Congo red by natural hydroxyapatite is 900 mg/g at pH 4.5, equilibrium time 240 min and adsorbent dosage 2 g/l. According to the correlation coefficient value, the pseudo-second order kinetic model identified the sorption mechanism of pollutant used on natural hydroxyapatite. The Freundlich isotherm is the best representative theoretical model of the dye molecules’ adsorption on the natural hydroxyapatite support.     

A potential trackable bone filler: Preparation and characterization

This study reported the synthesis of fluorescent hydroxyapatite/alginate/carbon quantum dots (HA/Alg/CQDs) nanocomposites via the co-precipitation technique. The N-doped CQDs as a new class of fluorescent materials were prepared by the citric acid pyrolysis method, with an average size around 4 nm. Physical, chemical, and optical properties of the synthesized nanocomposites were investigated by X-ray diffraction (XRD), Fourier-transformed infrared spectroscopy (FTIR), atomic force microscopy (AFM), field-emission scanning electron microscopy (FESEM), UV–visible spectroscopy, and photoluminescence (PL) spectroscopy, respectively. The PL spectroscopy data verified the favorable in vitro luminescent emission of the HA/Alg/CQDs nanocomposites in comparison with HA/Alg and HA samples. The XRD patterns of the prepared samples confirmed the formation of crystalline HA in all composites, possessing a Ca/P ratio around 1.5 as obtained by EDX elemental analysis. The FESEM analysis exhibited HA nanoplates that homogeneously distributed throughout the alginate matrix. Therefore, the synthesized nanocomposites could be regarded as potential trackable drug carriers for hard tissue engineering applications.     

Ca-P仿生人工骨材料的设计与制备

本文设计了用磷酸钙生物陶瓷制备仿生人工骨的工艺路线 ,从原料合成、材料成型和烧结等讨论了仿骨材料的制备方法     

生物陶瓷人工骨制备的新方法

主要介绍了近年来发展起来的两种新技术 :快速原型制造技术和放电等离子烧结技术。概括了两种技术的原理、特点以及在人工骨制备中的应用 ,提出了用这两种技术制备人工骨的设想。     

三种骨灰瓷原料的制备与晶相结构

本实验采用牛、猪、羊三种骨头通过高温煅烧制备骨灰瓷原料。采用XRD和SEM方法表征三种骨灰料的晶相结构和显微结构。实验结果表明 :三种骨灰料经 14 0 0℃煅烧后均存在Ca1 0 (PO4 ) 6(OH) 2 、α -Ca3(PO4 ) 2 种晶相。在 132 0℃煅烧的三种骨灰料中 ,按牛、猪、羊顺序 ,Ca1 0 (PO4 ) 6 (OH) 2 含量分别为 87 2 %、89 1%、71 9% ;β-Ca3(PO4 ) 2 含量最高 ,其可塑性和成瓷性能好 ,从晶相上考虑选择羊骨为较佳的骨灰瓷原料。从白度和透光度上考虑选择牛骨为较佳原料。随着煅烧温度升高 ,三种骨头料的β-Ca3(PO4 ) 2 含量增加 ,羊骨和牛骨中的Ca1 0 (PO4 ) 6 (OH) 2 含量减少 ,而猪骨的Ca1 0(PO4 ) 6 (OH) 2 含量反而增加。因此煅烧温度升高对羊骨和牛骨有利 ,而对猪骨不利     

骨质瓷生产经验浅谈

从骨质瓷的生产实际出发，并结合作者的生产经验概述了骨质瓷生产工艺中应着重了解和解决的问题。     

抗感染重组合异种骨的生物相容性研究

目的 检测抗感染重组合异种骨（ARBX）产品的生物相容性,确保ARBX产品在临床上安全、有效。方法 根据卫生部制定的《生物材料和制品的生物学评价标准》及《相关生物材料和制品的企业标准 Q/JGY001-94》,进行了ARBX急性毒性试验、过敏性试验、熟原试验、骨内埋植试验、肌内埋植试验、溶血试验及细胞毒性试验。结果ARBX未引起小鼠急性毒性反应、豚鼠过敏反应及家兔热原反应。在兔骨内和小鼠肌内埋植,既未产生纤维包膜,也未引起淋巴细胞及炎性细胞浸润。对家兔红细胞溶血率为1.2％,无溶血现象。体外细胞毒性反应指标R=0.25/0,无细胞毒性。结论ARBX的生物相容性符合国家规定的生物材料和制品的生物学标准及企业标准,可试用于临床治疗相关疾病。