Pore Architecture Of Scaffolds Research Articles

Bio-functional hydroxyapatite-coated 3D porous polyetherketoneketone scaffold for enhanced osteogenesis and osteointegration in orthopedic applications.

Polyetherketoneketone (PEKK), a high-performance thermoplastic special engineering material, maintains bone-like mechanical properties and has received considerable attention in the biomedical field. The 3D printing technique enables the production of porous scaffolds with a honeycomb structure featuring precisely controlled pore size, porosity and interconnectivity, which holds significant potential for applications in tissue engineering. The ideal pore architecture of porous PEKK scaffolds has yet to be elucidated. Porous PEKK scaffolds with five pore sizes P200 (225 ± 9.8 μm), P400 (411 ± 22.1 μm), P600 (596 ± 23.4 μm), P800 (786 ± 24.2 μm) and P1000 (993 ± 26.0 μm) were produced by a 3D printer. Subsequently, the optimum pore size, the P600, for mechanical properties and osteogenesis was selected based on in vitro experiments. To improve the interfacial bioactivity of porous PEKK scaffolds, hydroxyapatite (HAp) crystals were generated via in situ biomimetic mineralization induced by the phase-transited lysozyme coating. Herein, a micro/nanostructured surface showing HAp crystals on PEKK scaffold was developed. In vitro and in vivo experiments confirmed that the porous PEKK-HAp scaffolds exhibited highly interconnected pores and functional surface structures that were favorable for biocompatibility and osteoinductivity, which boosted bone regeneration. Therefore, this work not only demonstrates that the pore structure of the P600 scaffold is suitable for PEKK orthopedic implants but also sheds light on a synergistic approach involving 3D printing and biomimetic mineralization, which has the potential to yield customized 3D PEKK-HAp scaffolds with enhanced osteoinductivity and osteogenesis, offering a promising strategy for bone tissue engineering.

Microporous Annealed Particle (MAP) Scaffold Pore Size Influences Mesenchymal Stem Cell Metabolism and Proliferation Without Changing CD73, CD90, and CD105 Expression Over Two Weeks.

Scaffold pore architecture is shown to influence stem cell fate through various avenues. It is demonstrated that microporous annealed particle (MAP) microgel diameter can be tuned to control scaffold pore size and, in turn, modulate mesenchymal stem cell (MSC) survivability, proliferation, metabolism, and migration, thereby enhancing bioactivity and guiding future applications of MAP for regenerative medicine.

Virtual Design of 3D-Printed Bone Tissue Engineered Scaffold Shape Using Mechanobiological Modeling: Relationship of Scaffold Pore Architecture to Bone Tissue Formation.

Large bone defects are clinically challenging, with up to 15% of these requiring surgical intervention due to non-union. Bone grafts (autographs or allografts) can be used but they have many limitations, meaning that polymer-based bone tissue engineered scaffolds (tissue engineering) are a more promising solution. Clinical translation of scaffolds is still limited but this could be improved by exploring the whole design space using virtual tools such as mechanobiological modeling. In tissue engineering, a significant research effort has been expended on materials and manufacturing but relatively little has been focused on shape. Most scaffolds use regular pore architecture throughout, leaving custom or irregular pore architecture designs unexplored. The aim of this paper is to introduce a virtual design environment for scaffold development and to illustrate its potential by exploring the relationship of pore architecture to bone tissue formation. A virtual design framework has been created utilizing a mechanical stress finite element (FE) model coupled with a cell behavior agent-based model to investigate the mechanobiological relationships of scaffold shape and bone tissue formation. A case study showed that modifying pore architecture from regular to irregular enabled between 17 and 33% more bone formation within the 4-16-week time periods analyzed. This work shows that shape, specifically pore architecture, is as important as other design parameters such as material and manufacturing for improving the function of bone tissue scaffold implants. It is recommended that future research be conducted to both optimize irregular pore architectures and to explore the potential extension of the concept of shape modification beyond mechanical stress to look at other factors present in the body.

Bioceramic scaffolds with triply periodic minimal surface architectures guide early-stage bone regeneration.

The pore architecture of porous scaffolds is a critical factor in osteogenesis, but it is a challenge to precisely configure strut-based scaffolds because of the inevitable filament corner and pore geometry deformation. This study provides a pore architecture tailoring strategy in which a series of Mg-doped wollastonite scaffolds with fully interconnected pore networks and curved pore architectures called triply periodic minimal surfaces (TPMS), which are similar to cancellous bone, are fabricated by a digital light processing technique. The sheet-TPMS pore geometries (s-Diamond, s-Gyroid) contribute to a 3‒4-fold higher initial compressive strength and 20%-40% faster Mg-ion-release rate compared to the other-TPMS scaffolds, including Diamond, Gyroid, and the Schoen's I-graph-Wrapped Package (IWP) in vitro. However, we found that Gyroid and Diamond pore scaffolds can significantly induce osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Analyses of rabbit experiments in vivo show that the regeneration of bone tissue in the sheet-TPMS pore geometry is delayed; on the other hand, Diamond and Gyroid pore scaffolds show notable neo-bone tissue in the center pore regions during the early stages (3-5 weeks) and the bone tissue uniformly fills the whole porous network after 7 weeks. Collectively, the design methods in this study provide an important perspective for optimizing the pore architecture design of bioceramic scaffolds to accelerate the rate of osteogenesis and promote the clinical translation of bioceramic scaffolds in the repair of bone defects.

Controlling scaffold conductivity and pore size to direct myogenic cell alignment and differentiation.

Skeletal muscle's combination of three‐dimensional (3D) anisotropy and electrical excitability is critical for enabling normal movement. We previously developed a 3D aligned collagen scaffold incorporating conductive polypyrrole (PPy) particles to recapitulate these key muscle properties and showed that the scaffold facilitated enhanced myotube maturation compared with nonconductive controls. To further optimize this scaffold design, this work assessed the influence of conductive polymer incorporation and scaffold pore architecture on myogenic cell behavior. Conductive PPy and poly(3,4‐ethylenedioxythiophene) (PEDOT) particles were synthesized and mixed into a suspension of type I collagen and chondroitin sulfate prior to directional freeze‐drying to produce anisotropic scaffolds. Energy dispersive spectroscopy revealed homogenous distribution of conductive PEDOT particles throughout the scaffolds that resulted in a threefold increase in electrical conductivity while supporting similar myoblast metabolic activity compared to nonconductive scaffolds. Control of freezing temperature enabled fabrication of PEDOT‐doped scaffolds with a range of pore diameters from 98 to 238 μm. Myoblasts conformed to the anisotropic contact guidance cues independent of pore size to display longitudinal cytoskeletal alignment. The increased specific surface area of the smaller pore scaffolds helped rescue the initial decrease in myoblast metabolic activity observed in larger pore conductive scaffolds while also promoting modestly increased expression levels of the myogenic marker myosin heavy chain (MHC) and gene expression of myoblast determination protein (MyoD). However, cell infiltration to the center of the scaffolds was marginally reduced compared with larger pore variants. Together these data underscore the potential of aligned and PEDOT‐doped collagen scaffolds for promoting myogenic cell organization and differentiation.

Mechanical and Biological Behaviour of 3D Printed PCL-Based Scaffolds Fabricated by Fused Deposition Modelling for Bone Tissue Engineering: A Review of Recent Advances

The imperative need for alternative approaches to organ transplantation, replacing or regenerating damaged tissues is the key driving force for the remarkable development in tissue engineering. It can be precious in saving people who suffer from the critical shortage of organ donation. Such a strategy can repair injured body parts and tissues by using biomaterials, cells, and bioactive agents. Even though numerous scaffold manufacturing techniques have been available for bone regeneration, the three-dimensional (3D) printing approach can provide scaffolding with delicate features that may not be obtainable in other manufacturing strategies. For instance, when a 3D printer is used, it is possible to easily adjust scaffold pore architecture and size, porosity, and material alignment, forming customizable and defect-fillable scaffolds, which helps control the mechanical behavior of cellular response. The most prominent material used in scaffolding and printing is polycaprolactone (PCL), owing to its considerable potential and capabilities. It has favorable properties for the fabrication of bone tissue engineering scaffolds, such as biocompatibility, viscoelasticity, and affordability. Nonetheless, some inherent drawbacks of this polymer that limit its use in this field are detected, including inadequate mechanical performance, cell adhesion, osteoinductive deficiency, hydrophobicity, and low degradation rate. The incorporation of other materials within this polymer to form composites, on the other hand, can contribute to alleviating the negative influence of the PCL's undesirable characteristics. Improving the mechanical and biological behaviors of PCL-based scaffolds allows these structures to be utilized for tissue engineering since such composites can promote cell adhesion and differentiation, mimic anatomical characteristics of native bone, and can have superior mechanical performance. In this review, the latest advancements in printing intricate geometries 3D PCL-based composites using bioactive ceramics and/or biopolymers by fused deposition modeling (FDM) for bone tissue engineering will be explored, particularly from a morphological, mechanical, and biological perspective.

Bilayered extracellular matrix derived scaffolds with anisotropic pore architecture guide tissue organization during osteochondral defect repair

While some clinical advances in cartilage repair have occurred, osteochondral (OC) defect repair remains a significant challenge, with current scaffold-based approaches failing to recapitulate the complex, hierarchical structure of native articular cartilage (AC). To address this need, we fabricated bilayered extracellular matrix (ECM)-derived scaffolds with aligned pore architectures. By modifying the freeze-drying kinetics and controlling the direction of heat transfer during freezing, it was possible to produce anisotropic scaffolds with larger pores which supported homogenous cellular infiltration and improved sulfated glycosaminoglycan deposition. Neo-tissue organization in vitro could also be controlled by altering scaffold pore architecture, with collagen fibres aligning parallel to the long-axis of the pores within scaffolds containing aligned pore networks. Furthermore, we used in vitro and in vivo assays to demonstrate that AC and bone ECM derived scaffolds could preferentially direct the differentiation of mesenchymal stromal cells (MSCs) towards either a chondrogenic or osteogenic lineage respectively, enabling the development of bilayered ECM scaffolds capable of spatially supporting unique tissue phenotypes. Finally, we implanted these scaffolds into a large animal model of OC defect repair. After 6 months in vivo, scaffold implantation was found to improve cartilage matrix deposition, with collagen fibres preferentially aligning parallel to the long axis of the scaffold pores, resulting in a repair tissue that structurally and compositionally was more hyaline-like in nature. These results demonstrate how scaffold architecture and composition can be spatially modulated to direct the regeneration of complex interfaces such as the osteochondral unit, enabling their use as cell-free, off-the-shelf implants for joint regeneration. Statement of significanceThe architecture of the extracellular matrix, while integral to tissue function, is often neglected in the design and evaluation of regenerative biomaterials. In this study we developed a bilayered scaffold for osteochondral defect repair consisting of tissue-specific extracellular matrix (ECM)-derived biomaterials to spatially direct stem/progenitor cell differentiation, with a tailored pore microarchitecture to promote the development of a repair tissue that recapitulates the hierarchical structure of native AC. The use of this bilayered scaffold resulted in improved tissue repair outcomes in a large animal model, specifically the ability to guide neo-tissue organization and therefore recapitulate key aspects of the zonal structure of native articular cartilage. These bilayer scaffolds have the potential to become a new therapeutic option for osteochondral defect repair.

Bioceramic hydroxyapatite-based scaffold with a porous structure using honeycomb as a natural polymeric Porogen for bone tissue engineering

BackgroundThe application of bioceramic hydroxyapatite (HA) derived from materials high in calcium to tissue engineering has been of concern, namely scaffold. Scaffold pores allow for cell mobility metabolic processes, and delivery of oxygen and nutrients by blood vessel. Thus, pore architecture affects cell seeding efficiency, cell viability, migration, morphology, cell proliferation, cell differentiation, angiogenesis, mechanical strength of scaffolds, and, eventually, bone formation. Therefore, to improve the efficacy of bone regeneration, several important parameters of the pore architecture of scaffolds must be carefully controlled, including pore size, geometry, orientation, uniformity, interconnectivity, and porosity, which are interrelated and whose coordination affects the effectiveness of bone tissue engineering. The honeycomb (HCB) as natural polymeric porogen is used to pore forming agent of scaffolds. It is unique for fully interconnected and oriented pores of uniform size and high mechanical strength in the direction of the pores. The aim of this study was therefore to evaluate the effect of HCB concentration on macropore structure of the scaffolds.MethodsBioceramic hydroxyapatite (HA) was synthesized from abalone mussel shells (Halioitis asinina) using a precipitation method, and HA-based scaffolds were fabricated with honeycomb (HCB) as the porogen agent. Pore structure engineering was successfully carried out using HCB at concentrations of 10, 20, and 30 wt%.ResultsThe Energy Dispersive X-Ray Spectroscopy (EDS) analysis revealed that the Ca/P molar ratio of HA was 1.67 (the stoichiometric ratio of HA). The Fourier Transform Infrared Spectroscopy (FTIR) spectra results for porous HA-based scaffolds and synthesized HA showed that no chemical decomposition occurred in the HA-based scaffold fabrication process. The porosity of the scaffold tended to increase when higher concentrations of HCB were added. XRD data show that the HCB was completely degraded from the scaffold material. The cell metabolic activity and morphology of the HA + HCB 30 wt% scaffold enable it to facilitate the attachment of MC3T3E1 cells on its surface.ConclusionHCB 30 wt% is the best concentration to fabricate the scaffold corresponding to the criteria for pores structure, crystallographic properties, chemical decomposition process and cell viability for bone tissue engineering.

Digital light processing strength-strong ultra-thin bioceramic scaffolds for challengeable orbital bone regeneration and repair in Situ

The orbital fracture regeneration and reconstruction is still a catastrophic challenge due to the extremely thin anatomical structure and different defect shapes in various clinical conditions. The conventional biomaterials and preparation technique are often inefficient in satisfying porous scaffold architecture designs with respect to their required structural stability and biological performances. Here we developed the ultrathin scaffolds with high- precision pore structure and appreciable osteogenic activity for orbital bone repair via digital light processing (DLP) stereolithography. The dilute magnesium-substituting wollastonite (CSi-Mg) bioceramic scaffolds of ~1.5 mm in thickness and over 60% in porosity were investigated systematically, and compared with another calcium-magnesium silicate (i.e. akermanite) and the pure wollastonite porous counterparts. The results showed that the flexural strengths (~6−22 MPa) and in vitro biodegradation of such bioceramic scaffolds could be tuned during their chemical composition (i.e. Mg content) design stage, and the CSi-Mg scaffolds were also beneficial for osteogenic cell activity in vitro. In particular, the CSi-Mg scaffolds could readily enhance the new bone ingrowth at 4 weeks of implantation in a critical-sized rabbit orbital bone defect model, and such strength-strong scaffolds exhibited outstanding orbital defect repair efficacy after 12 weeks of implantation, in comparison with the structural instability of other scaffolds. Basically, it is demonstrated that the dilute magnesium doping in wollastonite bioceramic can be favorable for developing the mechanically reliable scaffolds with high porosity and bioactivity but quite thin wall morphology, and meanwhile the DLP-assisted additive manufacturing help us to optimize such porous bioceramic design with respect to their required structural stability for orbital bone reconstruction.

Bone tissue regeneration: The role of finely tuned pore architecture of bioactive scaffolds before clinical translation

Spatial dimension of pores and interconnection in macroporous scaffolds is of particular importance in facilitating endogenous cell migration and bone tissue ingrowth. However, it is still a challenge to widely tune structure parameters of scaffolds by conventional methods because of inevitable pore geometrical deformation and poor pore interconnectivity. Here, the long-term in vivo biological performances of nonstoichiometric bioceramic scaffolds with different pore dimensions were assessed in critical-size femoral bone defect model. The 6% Mg-substituted wollastonite (CSi-Mg6) powders were prepared via wet-chemical precipitation and the scaffolds elaborately printed by ceramic stereolithography, displaying designed constant pore strut and tailorable pore height (200, 320, 450, 600 μm), were investigated thoroughly in the bone regeneration process. Together with detailed structural stability and mechanical properties were collaboratively outlined. Both μCT and histological analyses indicated that bone tissue ingrowth was retarded in 200 μm scaffolds in the whole stage (2–16 weeks) but the 320 μm scaffolds showed appreciable bone tissue in the center of porous constructs at 6–10 weeks and matured bone tissue were uniformly invaded in the whole pore networks at 16 weeks. Interestingly, the neo-tissue ingrowth was facilitated in the 450 μm and 600 μm scaffolds after 2 weeks and higher extent of bone regeneration and remodeling at the later stage. These new findings provide critical information on how engineered porous architecture impact bone regeneration in vivo. Simultaneously, this study shows important implications for optimizing the porous scaffolds design by advanced additive manufacture technique to match the clinical translation with high performance.

Scaffold Architecture and Matrix Strain Modulate Mesenchymal Cell and Microvascular Growth and Development in a Time Dependent Manner.

Volumetric tissue-engineered constructs are limited in development due to the dependence on well-formed vascular networks. Scaffold pore size and the mechanical properties of the matrix dictates cell attachment, proliferation and successive tissue morphogenesis. We hypothesize scaffold pore architecture also controls stromal-vessel interactions during morphogenesis. The interaction between mesenchymal stem cells (MSCs) seeded on hydroxyapatite scaffolds of 450, 340, and 250μm pores and microvascular fragments (MVFs) seeded within 20mg/mL fibrin hydrogels that were cast into the cell-seeded scaffolds, was assessed in vitro over 21days and compared to the fibrin hydrogels without scaffold but containing both MSCs and MVFs. mRNA sequencing was performed across all groups and a computational mechanics model was developed to validate architecture effects on predicting vascularization driven by stiffer matrix behavior at scaffold surfaces compared to the pore interior. Lectin staining of decalcified scaffolds showed continued vessel growth, branching and network formation at 14days. The fibrin gel provides no resistance to spread-out capillary networks formation, with greater vessel loops within the 450μm pores and vessels bridging across 250μm pores. Vessel growth in the scaffolds was observed to be stimulated by hypoxia and successive angiogenic signaling. Fibrin gels showed linear fold increase in VEGF expression and no change in BMP2. Within scaffolds, there was multiple fold increase in VEGF between days 7 and 14 and early multiple fold increases in BMP2 between days 3 and 7, relative to fibrin. There was evidence of yap/taz based hippo signaling and mechanotransduction in the scaffold groups. The vessel growth models determined by computational modeling matched the trends observed experimentally. The differing nature of hypoxia signaling between scaffold systems and mechano-transduction sensing matrix mechanics were primarily responsible for differences in osteogenic cell and microvessel growth. The computational model implicated scaffold architecture in dictating branching morphology and strain in the hydrogel within pores in dictating vessel lengths.

Human trabecular meshwork cell behavior is influenced by collagen scaffold pore architecture and glycosaminoglycan composition.

Glaucoma is a degenerative eye disease in which damage to the optic nerve leads to a characteristic loss of vision. The primary risk factor for glaucoma is an increased intraocular pressure that is caused by an imbalance of aqueous humor generation and subsequent drainage through the trabecular meshwork (TM) drainage system. The small size, donor tissue limitations, and high complexity of the TM make it difficult to research the relationship between the TM cells and their immediate environment. Thus, a biomaterial-based approach may be more appropriate for research manipulations and in vitro drug development platforms. In this work, human TM (hTM) cells were cultured on various collagen scaffolds containing different glycosaminoglycans (GAGs) and different pore architectures to better understand how hTM cells respond to changes in their extracellular environment. Cellular response was measured by quantifying cellular proliferation and expression of an important extracellular matrix protein, fibronectin. The pore architecture of the scaffolds was altered using freeze-casting technique to make both large and small pores that were aligned or with a non-aligned random structure. The composition of the scaffolds was altered with the addition of chondroitin sulfate and/or hyaluronic acid. It was found that the hTM cells grown on large pore scaffolds proliferate more than those grown on small pores. There was an increase in the fibronectin expression with the incorporation of GAGs, and its morphology was changed by the underlying pore architecture. This work will help provide an insight into the behavior of hTM cells when introducing changes in their microenvironment.

Effects of macropore size in carbonate apatite honeycomb scaffolds on bone regeneration.

The pore architecture of scaffolds is a critical factor for angiogenesis and bone regeneration. Although the effects of scaffold macropore size have been investigated, most scaffolds feature macropores with poor uniformity and interconnectivity, and other parameters (e.g., microporosity, chemical composition, and strut thickness) differ among scaffolds. To clarify the threshold of effective macropore size, we fabricated honeycomb scaffolds (HCSs) with distinct macropore (i.e., channel) sizes (~100, ~200, and ~300μm). The HCSs were composed of AB-type carbonate apatite with ~8.5% carbonate ions, i.e., the same composition as human bone mineral. Their honeycomb architecture displayed uniformly sized and orderly arranged channels with extremely high interconnectivity, and all the HCSs displayed ~100-μm-thick struts and 0.06cm3g-1 of micropore volume. The compressive strengths of HCSs with ~100-, ~200-, and ~300-μm channels were higher than those of reported scaffolds, and decreased with increasing channel size: 62±6, 55±9, and 43±8MPa, respectively. At four weeks after implantation in rabbit femur bone defects, new bone and blood vessels were formed in all the channels of these HCSs. Notably, the ~300-μm channels were extensively occupied by new bone. We demonstrated that high interconnectivity and uniformity of channels can decrease the threshold of effective macropore size, enabling the scaffolds to maintain high mechanical properties and osteogenic ability and serve as implants for weight-bearing areas.

Physicochemical Characterization of Five Different Bone Graft Substitutes Used in Periodontal Regeneration: An In Vitro Study.

ABSTRACTBackground:Periodontal regeneration involves using a variety of bone graft substitutes (BGS) of varying origin and manufacturing processes. These include a wide range of biomaterials that are mainly of two types: the xenografts and alloplasts. The efficacy of these BGS depends upon the physical characteristics such as particle size, porous nature, surface morphology, as well as the chemical characteristics like composition, crystallinity and resorption properties.Aims:The present study is a descriptive study that focuses on describing the physicochemical characteristics of five selected commercially available BGS that are frequently used in periodontal regeneration procedures. The BGS studied here included two xenografts (colocast and osseograft) and three alloplasts (B-OstIN, biograft HABG active and biograft HT).Materials and Methods:The physical properties of the BGS, including particle size, morphology, and surface topography, were analyzed using SEM. The mineral phases and crystallinity of the BGS were analyzed using XRD.Results:The results showed that the xenografts (colocast and osseograft) had minimal mineral composition and crystalline structure. The physical properties such as surface roughness and porosity were less compared to alloplastic materials. The alloplasts (B-OstIN, biograft HABG and biograft HT) that had different chemical compositions showed varying physical and crystalline properties. Biograft HT showed a superior porous scaffold architecture among all BGS studied.Conclusion:It is important for a clinician to have a thorough understanding about the physicochemical characteristics of BGS they use in periodontal regeneration. The xenografts evaluated here had minimal physical and crystalline properties. Among the alloplasts studied, biograft HT showed superior physicochemical properties, while the presence of bioactive glass in biograft HABG enhanced regeneration.

Effect of Unit Cell Type and Pore Size on Porosity and Mechanical Behavior of Additively Manufactured Ti6Al4V Scaffolds.

Porous metal structures have emerged as a promising solution in repairing and replacing damaged bone in biomedical applications. With the advent of additive manufacturing technology, fabrication of porous scaffold architecture of different unit cell types with desired parameters can replicate the biomechanical properties of the natural bone, thereby overcoming the issues, such as stress shielding effect, to avoid implant failure. The purpose of this research was to investigate the influence of cube and gyroid unit cell types, with pore size ranging from 300 to 600 µm, on porosity and mechanical behavior of titanium alloy (Ti6Al4V) scaffolds. Scaffold samples were modeled and analyzed using finite element analysis (FEA) following the ISO standard (ISO 13314). Selective laser melting (SLM) process was used to manufacture five samples of each type. Morphological characterization of samples was performed through micro CT Scan system and the samples were later subjected to compression testing to assess the mechanical behavior of scaffolds. Numerical and experimental analysis of samples show porosity greater than 50% for all types, which is in agreement with desired porosity range of natural bone. Mechanical properties of samples depict that values of elastic modulus and yield strength decreases with increase in porosity, with elastic modulus reduced up to 3 GPa and yield strength decreased to 7 MPa. However, while comparing with natural bone properties, only cube and gyroid structure with pore size 300 µm falls under the category of giving similar properties to that of natural bone. Analysis of porous scaffolds show promising results for application in orthopedic implants. Application of optimum scaffold structures to implants can reduce the premature failure of implants and increase the reliability of prosthetics.

Pore size directs bone marrow stromal cell fate and tissue regeneration in nanofibrous macroporous scaffolds by mediating vascularization

In the U.S., 30% of adults suffer joint pain, most commonly in the knee, which severely limits mobility and is often attributed to injury of cartilage and underlying bone in the joint. Current treatment methods such as microfracture result in less resilient fibrocartilage with eventual failure; autografting can cause donor site morbidity and poor integration. To overcome drawbacks in treatment, tissue engineers can design cell-instructive biomimetic scaffolds using biocompatible materials as alternate therapies for osteochondral defects. Nanofibrous poly (l-lactic acid) (PLLA) scaffolds of uniform, spherical, interconnected and well-defined pore sizes that are fabricated using a thermally-induced phase separation and sugar porogen template method create an extracellular matrix-like environment which facilitates cell adhesion and proliferation. Herein we report that chondrogenesis and endochondral ossification of rabbit and human bone marrow stromal cells (BMSCs) can be controlled by scaffold pore architecture, particularly pore size. Small-pore scaffolds support enhanced chondrogenic differentiation in vitro and cartilage formation in vivo compared to large-pore scaffolds. Endochondral ossification is prevented in scaffolds with very small pore sizes; pore interconnectivity is critical to promote capillary ingrowth for mature bone formation. These results provide a novel strategy to control tissue regenerative processes by tunable architecture of macroporous nanofibrous scaffolds. Statement of significanceProgress in understanding the relationship between cell fate and architectural features of tissue engineering scaffolds is critical for engineering physiologically functional tissues. Sugar porogen template scaffolds have uniform, spherical, highly interconnected macropores. Tunable pore-size guides the fate of bone marrow stromal cells (BMSCs) towards chondrogenesis and endochondral ossification, and is a critical design parameter to mediate neotissue vascularization. Preventing vascularization favors a chondrogenic cell fate while allowing vascularization results in endochondral ossification and mineralized bone formation. These results provide a novel strategy to control tissue regenerative processes by tunable architecture of macroporous nanofibrous scaffolds.

Osteogenesis by foamed and 3D-printed nanostructured calcium phosphate scaffolds: Effect of pore architecture

There is an urgent need of synthetic bone grafts with enhanced osteogenic capacity. This can be achieved by combining biomaterials with exogenous growth factors, which however can have numerous undesired side effects, but also by tuning the intrinsic biomaterial properties. In a previous study, we showed the synergistic effect of nanostructure and pore architecture of biomimetic calcium deficient hydroxyapatite (CDHA) scaffolds in enhancing osteoinduction, i.e. fostering the differentiation of mesenchymal stem cells to bone forming cells. This was demonstrated by assessing bone formation after implanting the scaffolds intramuscularly. The present study goes one step forward, since it analyzes the effect of the geometrical features of the same CDHA scaffolds, obtained either by 3D-printing or by foaming, on the osteogenic potential and resorption behaviour in a bony environment. After 6 and 12 weeks of intraosseous implantation, both bone formation and material degradation had been drastically affected by the macropore architecture of the scaffolds. Whereas nanostructured CDHA was shown to be highly osteoconductive both in the robocast and foamed scaffolds, a superior osteogenic capacity was observed in the foamed scaffolds, which was associated with their higher intrinsic osteoinductive potential. Moreover, they showed a significantly higher cell-mediated degradation than the robocast constructs, with a simultaneous and progressive replacement of the scaffold by new bone. In conclusion, these results demonstrate that the control of macropore architecture is a crucial parameter in the design of synthetic bone grafts, which allows fostering both material degradation and new bone formation.Statement of Significance3D-printing technologies open new perspectives for the design of patient-specific bone grafts, since they allow customizing the external shape together with the internal architecture of implants. In this respect, it is important to design the appropriate pore geometry to maximize the bone healing capacity of these implants. The present study analyses the effect of pore architecture of nanostructured hydroxyapatite scaffolds, obtained either by 3D-printing or foaming, on the osteogenic potential and scaffold resorption in an in vivo model. While nanostructured hydroxyapatite showed excellent osteoconductive properties irrespective of pore geometry, we demonstrated that the spherical, concave macropores of foamed scaffolds significantly promoted both material resorption and bone regeneration compared to the 3D-printed scaffolds with orthogonal-patterned struts and therefore prismatic, convex macropores.

Influence of Stage Cooling Method on Pore Architecture of Biomimetic Alginate Scaffolds

Porous scaffold is widely used in the field of tissue engineering. However, the anisotropic structure of actual extracellular matrix (ECM) of human tissue pose a challenge to the scaffold structure that pore size should be changed in gradient. Here we report a stage cooling method to fabricate alginate scaffold with gradient pores. Eight cooling models were set according to different temperature steps, different initial temperature, and different time duration. The thermal characterization of solution during cooling process were recorded and scaffold morphology were observed. The results revealed that the temperature steps mainly affected pore shape, while the initial temperature and time duration mainly affected pore size. By altering the initial temperature and time duration, scaffold exhibited cellular and gradually enlarged pores on the vertical axial direction (10–65 μm at base, 50–141 μm at top). With this stage cooling method, pore shape and pore size could be easily tailored and scaffold with gradient structure could be fabricated.

Development of a synthetic tissue engineered three-dimensional printed bioceramic-based bone graft with homogenously distributed osteoblasts and mineralizing bone matrix in vitro.

Over the last decade there have been increasing efforts to develop three-dimensional (3D) scaffolds for bone tissue engineering from bioactive ceramics with 3D printing emerging as a promising technology. The overall objective of the present study was to generate a tissue engineered synthetic bone graft with homogenously distributed osteoblasts and mineralizing bone matrix in vitro, thereby mimicking the advantageous properties of autogenous bone grafts and facilitating usage for reconstructing segmental discontinuity defects in vivo. To this end, 3D scaffolds were developed from a silica-containing calcium alkali orthophosphate, using, first, a replica technique - the Schwartzwalder-Somers method - and, second, 3D printing, (i.e. rapid prototyping). The mechanical and physical scaffold properties and their potential to facilitate homogenous colonization by osteogenic cells and extracellular bone matrix formation throughout the porous scaffold architecture were examined. Osteoblastic cells were dynamically cultured for 7 days on both scaffold types with two different concentrations of 1.5 and 3 × 109 cells/l. The amount of cells and bone matrix formed and osteogenic marker expression were evaluated using hard tissue histology, immunohistochemical and histomorphometric analysis. 3D-printed scaffolds (RPS) exhibited more micropores, greater compressive strength and silica release. RPS seeded with 3 × 109 cells/l displayed greatest cell and extracellular matrix formation, mineralization and osteocalcin expression. In conclusion, RPS displayed superior mechanical and biological properties and facilitated generating a tissue engineered synthetic bone graft in vitro, which mimics the advantageous properties of autogenous bone grafts, by containing homogenously distributed terminally differentiated osteoblasts and mineralizing bone matrix and therefore is suitable for subsequent in vivo implantation for regenerating segmental discontinuity bone defects. Copyright © 2016 John Wiley & Sons, Ltd.

A bioprosthetic ovary created using 3D printed microporous scaffolds restores ovarian function in sterilized mice

Emerging additive manufacturing techniques enable investigation of the effects of pore geometry on cell behavior and function. Here, we 3D print microporous hydrogel scaffolds to test how varying pore geometry, accomplished by manipulating the advancing angle between printed layers, affects the survival of ovarian follicles. 30° and 60° scaffolds provide corners that surround follicles on multiple sides while 90° scaffolds have an open porosity that limits follicle–scaffold interaction. As the amount of scaffold interaction increases, follicle spreading is limited and survival increases. Follicle-seeded scaffolds become highly vascularized and ovarian function is fully restored when implanted in surgically sterilized mice. Moreover, pups are born through natural mating and thrive through maternal lactation. These findings present an in vivo functional ovarian implant designed with 3D printing, and indicate that scaffold pore architecture is a critical variable in additively manufactured scaffold design for functional tissue engineering.