磷酸钙生物陶瓷的孔性、内部连通性及表面形态对骨质再生的影响

磷酸钙生物陶瓷是一类具有良好生物相容性和骨传导性的生物活性材料,由于其成分与骨的无机组分基本相当,是目前公认的最具前途的硬组织修复材料,将该材料制成多孔结构,更有利于新生骨组织生长所需的物质交流,促进新生骨形成,因而成为近年来生物材料的研究热点之一。本文从多孔磷酸钙生物陶瓷与组织的响应关系角度出发,评述了近年来磷酸钙生物陶瓷的孔性、孔和孔间的内部连通性及孔的表面形态对骨质再生影响的研究进展。     

Nickel release behavior, cytocompatibility, and superelasticity of oxidized porous single-phase NiTi

Porous NiTi shape memory alloys are one of the promising biomaterials for surgical implants because of their unique shape memory effects and porous structure with open pores. However, the complex surface morphology and larger area of porous NiTi compared to dense NiTi make it more vulnerable from the viewpoint of release of nickel, which can cause deleterious effects in the human body. It is also more difficult to modify the exposed surfaces of a porous structure using conventional surface modification technologies. In this work, oxidation in conjunction with postreaction heat treatment was used to modify the surfaces of porous single-phase NiTi prepared by capsule-free hot isostatic pressing to mitigate Ni leaching and enhance the surface properties. Differential scanning calorimetry thermal analysis, uniaxial compression tests, inductively-coupled plasma mass spectrometry, and cell cultures reveal that porous NiTi alloys oxidized at 450 degrees C for 1 h have an austenite transition temperature below 37 degrees C, excellent superelasticity, lower nickel release, and no cytotoxicity.     

组织工程用聚合物多孔支架的制备技术

组织工程用多孔支架不仅要有普通生物材料具有的性能,而且对其孔径、孔隙率、比表面积等物理性能也有一定的要求,而这些物理性能与多孔支架的制备工艺密切相关。本文对组织工程用聚合物多孔支架的制备方法进行了较为全面的回顾,包括纤维粘结法、粒子致孔法、熔融成型法、气体发泡法、相分离法、烧结微球法和快速成型等方法。每种方法各有其优缺点,将多种方法结合,制备出同时具有复杂外形和规则的相连孔结构是组织工程多孔支架制备技术研发的方向。     

Multinozzle low-temperature deposition system for construction of gradient tissue engineering scaffolds

Tissue engineering is a technology that enables us to construct complicated hominine organs composed of many different types of cells. One of the key points to achieve this goal is to control the material composition and porous structure of the scaffold accurately. A disposable syringe based volume-driven injecting (VDI) nozzle was proposed and designed to extrude both natural derived and synthetic polymers. A multinozzle low-temperature deposition and manufacturing (M-LDM) system is proposed to fabricate scaffolds with heterogeneous materials and gradient hierarchical porous structures. PLGA, collagen, gelatin, chitosan can be extruded without leaking to form hierarchical porous scaffolds for primary study. Composite scaffolds with two kinds of materials were fabricated via two different nozzles to get both hydrophilic and mechanical properties. The results from scanning electron microscopy (SEM) demonstrated that the natural-derived biomaterials were strongly absorbed onto the synthetic biomaterials to form a stable network. Several gradient PLGA/TCP scaffolds were also fabricated to supply several samples.     

Interactions of acinar cells on biomaterials with various surface properties

The purpose of this study is to evaluate the interactions of rat parotid acinar cells on biomaterials with different surface properties. The biomaterials used in this study included polyvinyl alcohol (PVA), chitosan, poly (ethylene-co-vinyl alcohol) (EVAL), and polyvinylidene fluoride (PVDF). Cell morphology was observed by photomicroscope. Cell growth and differentiated characteristic function were separately assayed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction activity and amylase activity. Results indicated that behaviors of acinar cells on materials might differ to a great extent depending on the surface hydrophilicity and morphology of the materials. On the relatively hydrophobic materials, the abilities of acinar cells to adhere and proliferate increased simultaneously. In addition, porous PVDF had higher cell growth compared with dense PVDF. Therefore, the hydrophobic PVDF with a porous structure was the best substrate for culturing acinar cells. According to our findings, a tubular PVDF scaffold with dense outer surface to prevent saliva leakage and with porous inner surface for the cell growth was proposed to serve as an artificial salivary gland for future use in the treatment of patients with salivary hypofunction.     

Fabrication of Chitosan Scaffolds with Tunable Porous Orientation Structure for Tissue Engineering

Chitosan, as an example of natural macromolecular biomaterials, was used to fabricate highly porous chitosan scaffolds with microtubules having a tubular orientation structure using the unidirectional freeze-drying method. The porous structure of the scaffolds was characterized via scanning electron microscopy. The factors that affect the porous structure of the scaffolds, such as the concentration of chitosan solution and addition of glutaraldehyde as cross-linking agent, have been extensively studied in order to find a facile and efficient way to control the porosity, tubular morphology and orientation of the microtubules. The properties of the chitosan scaffolds, including water absorption ability, compressive strength, protein adsorption and in vitro enzymatic biodegradation in the presence of lysozyme, were also investigated. In vitro cell-culture results showed that the chitosan scaffold was non-toxic to cartilage cells and the cells could spread and grow well on the scaffolds.     

Comparative in vitro study of the proliferation and growth of human osteoblast-like cells on various biomaterials

In vitro studies about the growth behavior of osteoblasts onto biomaterials is a basic knowledge and a screening method for the development and application of scaffolds in vivo. In this in vitro study human osteoblast-like (HOB) cells were cultured on seven different biomaterials used in dental and craniomaxillofacial surgery, respectively. The tested biomaterials were synthetic biodegradable (MacroPore, Ethisorb, PDS, Beriplast P) and nonbiodegradable polymers (Palacos) as well as calcium phosphate cement (BoneSource) and titanium. The cell proliferation and cell colonization were analyzed by scanning electron microscopy and EZ4U-test. Statistical analysis were performed. HOB-like cells cultivated on Ethisorb showed the highest proliferation rate. The proliferation rate was statistically significant compared with Palacos, MacroPore, and BoneSource. Whereas, Beriplast, PDS, and titanium yielded lower proliferation rates. However, there was no statistically significant difference compared with Palacos, MacroPore, and BoneSource. SEM analysis showed no significant difference in individual cell features and cell colonization. But an infiltration and a growth of HOB-like cells throughout the porous structure of Ethisorb, which is formed by crossing fibers, is a striking different feature (macrotopography). This feature can explain the highest proliferation rate of Ethisorb. The results showed that HOB-like cells appear to be sensitive to substrate composition and topography. Moreover, the basis for further studies with such biomaterial/osteoblast constructs in vivo are provided. Further focusing points are developing techniques to fabricate three-dimensional porous biomaterial/cell constructs, studying the tissue reaction and the bone regeneration of such constructs compared with the use of autologous bone.     

Porous TiNbZr alloy scaffolds for biomedical applications

In the present study, porous Ti–10Nb–10Zr alloy scaffolds with different porosities were successfully fabricated by a “space-holder” sintering method. By the addition of biocompatible alloying elements the porous TiNbZr scaffolds achieved significantly higher strength than unalloyed Ti scaffolds of the same porosity. In particular, the porous TiNbZr alloy with 59% porosity exhibited an elastic modulus and plateau stress of 5.6 GPa and 137 MPa, respectively. The porous alloys exhibited excellent ductility during compression tests and the deformation mechanism is mainly governed by bending and buckling of the struts. Cell cultures revealed that SaOS2 osteoblast-like cells grew on the surface and inside the pores and showed good spreading. Cell viability for the porous scaffold was three times higher than the solid counterpart. The present study has demonstrated that the porous TiNbZr alloy scaffolds are promising scaffold biomaterials for bone tissue engineering by virtue of their appropriate mechanical properties, highly porous structure and excellent biocompatibility.     

新型药物载体——聚/假聚氨基酸

从分子水平设计 ,以具有营养和药理功能的精、赖和天冬氨基酸为单体 ,合成聚合和 /或假型聚合多肽高分子材料 ,制成生物降解产物不仅是可吸收而且是介入治疗的新型医用薄膜、多孔体和微球用作药物载体。该材料所具有特性便于探讨材料的结构、代谢途径和对机体的影响以及载体与药物之间相互关系和机理     

Scaffolds based on hyaluronan crosslinked with a polyaminoacid: Novel candidates for tissue engineering application

New porous scaffolds, with a suitable hydrolytic and enzymatic degradation, useful for tissue engineering applications have been obtained by a carbodiimide mediated reaction between hyaluronan (HA) and a synthetic polymer with a polyaminoacid structure such as alpha,beta-polyaspartylhydrazide (PAHy). Scaffolds with a different molar ratio between PAHy repeating units and HA repeating units have been prepared and characterized from a chemical and physicochemical point of view. Tests of indirect and direct cytotoxicity, cell adhesion, and spreading on these biomaterials have been performed by using murine L929 fibroblasts. The new biomaterials showed a good cell compatibility and ability to allow cell migration into the scaffolds as well as spreading on their surface.     

3D imaging of tissue integration with porous biomaterials

Porous biomaterials designed to support cellular infiltration and tissue formation play a critical role in implant fixation and engineered tissue repair. The purpose of this Leading Opinion Paper is to advocate the use of high resolution 3D imaging techniques as a tool to quantify extracellular matrix formation and vascular ingrowth within porous biomaterials and objectively compare different strategies for functional tissue regeneration. An initial over-reliance on qualitative evaluation methods may have contributed to the false perception that developing effective tissue engineering technologies would be relatively straightforward. Moreover, the lack of comparative studies with quantitative metrics in challenging pre-clinical models has made it difficult to determine which of the many available strategies to invest in or use clinically for companies and clinicians, respectively. This paper will specifically illustrate the use of microcomputed tomography (micro-CT) imaging with and without contrast agents to nondestructively quantify the formation of bone, cartilage, and vasculature within porous biomaterials.     

Osteoinduction of porous Ti implants with a channel structure fabricated by selective laser melting

Many studies have shown that certain biomaterials with specific porous structures can induce bone formation in non-osseous sites without the need for osteoinductive biomolecules, however, the mechanisms responsible for this phenomenon (intrinsic osteoinduction of biomaterials) remain unclear. In particular, to our knowledge the type of pore structure suitable for osteoinduction has not been reported in detail. In the present study we investigated the effects of interconnective pore size on osteoinductivity and the bone formation processes during osteoinduction. Selective laser melting was employed to fabricate porous Ti implants (diameter 3.3mm, length 15 mm) with a channel structure comprising four longitudinal square channels, representing pores, of different diagonal widths, 500, 600, 900, and 1200 μm (termed p500, p600, p900, and p1200, respectively). These were then subjected to chemical and heat treatments to induce bioactivity. Significant osteoinduction was observed in p500 and p600, with the highest observed osteoinduction occurring at 5mm from the end of the implants. A distance of 5mm probably provides a favorable balance between blood circulation and fluid movement. Thus, the simple architecture of the implants allowed effective investigation of the influence of the interconnective pore size on osteoinduction, as well as the relationship between bone quantity and its location for different pore sizes.     

Micro-finite element models of bone tissue-engineering scaffolds

Tissue engineering is an emerging area in bioengineering at the frontiers between biomaterials, biology and biomechanics. The basic knowledge of the interactions between mechanical stimuli, cells and biomaterials is growing but the quantitative effect of mechanical stimuli on cells attached to biomaterials is still unknown. The objective of this study was to develop finite element models of various bone scaffolds based on calcium phosphate in order to calculate the load transfer from the biomaterial structure to the biological entities. Samples of porous calcium phosphate bone cement and biodegradable glass were scanned using micro-CT to determine the overall macroporosity, architecture and to develop finite element models of such materials. Compressive loads were applied on the models to simulate the in vitro environment of a bioreactor and stress and strain distributions were calculated. It was found that the effective Young's modulus was linearly related to the sample macroporosity. Results suggest that a 0.5% overall compressive strain can produce internal strain of the same order of magnitude as found in previous in vitro mechanically cell-strained studies or in mechanoregulation studies. Stress and strain concentrations due to the porous structures are possible candidate for favouring cell differentiation. Although strain distributions were similar between bone cement and porous glass, the stress distribution is clearly different. Future in vitro results could correlate the results obtained with such finite element study to explain the influence of mechanical stimuli on cell behaviour.     

Tissue-like self-assembly in cocultures of endothelial cells and osteoblasts and the formation of microcapillary-like structures on three-dimensional porous biomaterials

The survival and functioning of a bone biomaterial requires a rapid and stable vascularization after implantation. However, the mechanisms involved in the context of the complex healing microenvironment are poorly understood. To evaluate the vascularization potential of bone biomaterials, angiogenic stimuli were added to human dermal microvascular endothelial cells (HDMEC) growing on three-dimensional (3-D) bone biomaterials consisting of porous hydroxyapatite, porous calcium phosphate, porous nickel-titanium, successfully being used in humans, and also silk fibroin nets. HDMEC did not migrate to form microcapillary-like structures as they did on cell culture plastic. In cocultures of HDMEC and primary human osteoblast cells (HOS) or the human osteoblast-like cell line MG-63 on these biomaterials, a tissue-like self-assembly of cells occurred with time, with endothelial cells forming microcapillary-like structures containing a lumen and giving a strong PECAM-1 expression at cell interfaces. These microcapillary-like structures were intertwined between cell layers of osteoblasts and did not form when exogenous angiogenic stimuli were added to these cocultures. The life span of HDMEC was also significantly enhanced by coculture; with HDMEC being present for up to at least 42 days, compared to the monoculture where cells began to die rapidly after 1 week without passage. This coculture system may be applicable to a prevascularization strategy for biomaterials prior to implantation. Irrespective of this, the coculture model holds promise for studies to deepen our understanding of bone regeneration on 3-D substrates. Most importantly, these data raise important questions concerning the exact nature of pro-angiogenic drug- or gene-delivery systems to be incorporated into scaffolds. Our results underline the necessity to take into account the in situ production of growth factors by invading mesenchymal cells in the regenerative niche.     

Impact of sterilization on the porous design and cell behavior in collagen sponges prepared for tissue engineering

This study investigates the impact of different sterilization processes on structural integrity and stability of collagen sponges designed for tissue engineering. Collagen sponges with uniform pore size (20 microm) were sterilized either with ethylene oxide (EO) or gamma irradiation (2.5 Mrad). Gamma-sterilized sponges showed a dramatic decrease of resistance against enzyme degradation and severe shrinkage after cell seeding. Collapsed porosity inhibited fibroblasts and barred completely the human umbilical vein endothelial cell ingrowth into the sponges. On the contrary, the porous structure and stability of EO-sterilized sponges remained almost unaltered. Fibroblasts and endothelial cells exhibited favorable proliferation and migration within sponges with normal morphology. Tubular formation by seeded endothelial cells occurred early in the first week. Therefore, we emphasize that the impact of sterilization of biomaterials is substantial and any new procedure has to be evaluated by correlating the impact of the procedure on the porous structure with cell proliferation behavior.     

Pore throat size and connectivity determine bone and tissue ingrowth into porous implants: three-dimensional micro-CT based structural analyses of porous bioactive titanium implants

A porous structure comprises pores and pore throats with a complex three-dimensional (3D) network structure, and many investigators have described the relationship between average pore size and the amount of bone ingrowth. However, the influence of network structure or pore throats for tissue ingrowth has rarely been discussed. Four types of bioactive porous titanium implants with different pore sizes and porosities (6mm in diameter and 15 mm long) were analyzed using specific algorithms for 3D analysis of interconnectivity based on a micro focus X-ray computed tomography system. In vivo histomorphometric analysis was performed using the very same implants implanted into the femoral condyles of male rabbits for 6 and 12 weeks. This matching study revealed that more poorly differentiated pores tended to have narrow pore throats, especially in their shorter routes to the outside. In addition, for assessment of the entire implant, we proposed new two indices that represent the degree of bone and tissue ingrowth into an implant by considering the effect of narrow pore throats. Data obtained suggest that this sort of novel analysis is useful for evaluating bone and tissue ingrowth into porous biomaterials.     

Modulation of porosity in apatitic cements by the use of alpha-tricalcium phosphate-calcium sulphate dihydrate mixtures

Calcium phosphate bone cements are injectable biomaterials that are being used in dental and orthopaedic applications through minimally invasive surgery techniques. Nowadays, apatitic bone cements based on alpha-tricalcium phosphate (alpha-TCP) are of special interest due to their self-setting behaviour when mixed with an aqueous liquid phase. In this study, a new method to improve osteointegration of alpha-TCP-based cements is presented. This method consists in the modification of the cement's powder phase with different amounts of calcium sulphate dihydrate (CSD). The resulting hardening properties of the new biphasic cements are a combination between the progressive hardening due to the main alpha-TCP reactant and the progressive dissolution of the CSD phase, which render a porous material. It was observed that the maximum compressive strength of Biocement-H (45 MPa) decreased as the amount of CSD increased in the cement powder mixture ( approximately 30 MPa for 25 wt% of CSD). It was also observed that after complete dissolution of the CSD phase a porous apatitic structure appears with a mechanical compressive strength suitable for cancellous bone applications (10 MPa).     

Biomaterials science protocols for clinical investigations on porous alumina ceramic and vitreous carbon implants.

A written protocol for the investigation of candidate surgical implant materials is quite important. Biomaterials science sections of clinical protocols have been developed for porous alumina ceramic and nonporous vitreous carbon biomaterials. Published data on the properties of the biomaterials were evaluated as related to bone replacement and augmentation. Where necessary, limited laboratory studies were conducted. If decisions could not be reached with respect to a given application, animal studies were initiated. The surgeons worked with biomaterials in the laboratory and the biomaterials scientist attended the experimental surgery procedures. Biomaterials Science Laboratory nondestructive investigations including stereomicroscopic and x-ray inspections were conducted on the vitreous carbon dental implant systems. The investigations elucidated a number of unexpected features for both implant biomaterials and the overall interaction between the different disciplines resulted in a more complete protocol for the study of these biomaterials at our Medical and Dental Center.     

天然生物材料构建组织工程支架的研究进展

细胞培养支架材料是组织工程的重要研究内容之一.本文综述近年来几种典型天然生物材料构建组织工程支架的研究进展,并详细介绍了适用于天然生物材料制备组织工程支架的致孔方法.     

Nanofibrous scaffold from electrospinning biodegradable waterborne polyurethane/poly(vinyl alcohol) for tissue engineering application

A series of nanofibrous scaffolds, free of organic solvents, are prepared by electrospinning biodegradable waterborne polyurethane (BWPU) emulsion blending with aqueous poly(vinyl alcohol)(PVA). Tuning the proration of BWPU to PVA, various nanofibers with diameter from 370 to 964 nm are obtained. Strong intermolecular interaction existing between them benefits to the electrospun of BWPU emulsion, which is demonstrated by dynamic thermomechanical analysis and Fourier transform infrared spectroscopy. The nontoxic nanofibrous scaffolds with porous structure, which is similar to the natural extracellular matrix, favor to the attachment and proliferation of the L929 fibroblasts. Thus, the scaffolds are promising to be used as biomaterials for many natural tissues repair.