The significant impact of mechanically-induced phase transformation on cellular functionality of biomedical austenitic stainless steel.

The implant surface and tissue experience strain when micro-motion occurs at the bone-implant interface under physiological loading. Moreover, strain is also introduced on the surface during mechanical processing of biomedical devices. Both these situations can induce phase transformation depending on the degree of stability of the microstructural constituents. In this regard, we elucidate here the interplay between mechanically-induced phase transformation (strain-induced martensite) in austenitic stainless steel on osteoblast functions. Strain-induced martensite significantly impacted cellular functions, notably, cell attachment, cell-surface interactions, proliferation, and synthesis of prominent proteins (fibronectin, actin, and vinculin). Strain-induced martensite favorably modulated cellular activity and contributed to small differences in hydrophilicity in relation to the non-strained austenitic stainless steel surface. The study provides a pathway for tuning biological functionality via microstructural control facilitated by mechanical strain.

Bone regeneration of hollow tubular magnesium­strontium scaffolds in critical-size segmental defects: Effect of surface coatings.

Segmental defects are formidable challenges for orthopedic surgeons that are caused by large osseous defects such as open injury, comminuted fracture as well as other severe traumas and infection. Current treatment options have practical and clinical shortcomings, calling for innovative bone graft materials. This study is related to hollow tubular magnesium­strontium (MgSr) alloy scaffolds with autologous morselized bone filled inside and three different coatings were individually applied on MgSr scaffolds, respectively, to study the effects of degradation and bioactivity of the grafts on new bone growth. The optimal coating method was screened using immersion tests, cell proliferation and adhesion, alkaline phosphatase (ALP) assay in vitro, and 4 weeks' implantation in a critical-size segmental defect in vivo. More new bone formation was observed by radiographic tests and histology along the ulna defects, when magnesium scaffold grafts were implanted. Meanwhile, depression occurred for blank control group with only autologous morselized bone filled, because of rapid absorption rate of morselized bone during initial implantation. Therefore, biodegradable MgSr alloy grafts showed their potential application in treating the critical-size segmental defects. As for different coating methods, CaP chemically deposited (CaP) coated sample showed least H2 evolution in vivo, demonstrating highest corrosion resistance and relative stable interfaces, however, the least beneficial ion release meanwhile. Micro-arc oxidation coating (MAO) degraded faster comparing with CaP, while with the main composition of MgO. They both indicate insufficient bioactivity in bone formation. The results suggest superior combination of bioactive surface, beneficial ions release and appropriate corrosion rate of Strontium phosphate conversion (SrP) coating, indicating superior comprehensive oeteoconductive and osteoinductive properties of coatings on hollow tubular MgSr alloy scaffold.

The determining role of nanoscale mechanical twinning on cellular functions of nanostructured materials.

Considering that micromotions generated at the bone-implant interface under physiological loading introduce mechanical strain on the tissue and surface of the implant and that strain can be introduced during processing of the biomedical device, we elucidate here the interplay between mechanically-induced nanoscale twinning in austenitic stainless steel on osteoblast functions. Mechanically-induced nanoscale twinning significantly impacted cell attachment, cell-substrate interactions, proliferation, and subsequent synthesis of prominent proteins (fibronectin, actin, and vinculin). Twinning was beneficial in favorably modulating cellular activity and contributed to small differences in hydrophilicity and nanoscale roughness in relation to the untwinned surface.

Alginate/poly(amidoamine) injectable hybrid hydrogel for cell delivery.

A covalently cross-linked injectable hybrid hydrogel, namely, alginate/poly(amidoamine) (PAMAM), with the objective of cell delivery was innovatively designed and synthesized using tetra-amino-functional PAMAM dendrimer as the cross-linker. With the increase in percentage of PAMAM cross-linker, the pore size and swelling ratio of hydrogels were in the range of 57 ± 18 µm to 88 ± 25 µm and 110 ± 16 to 157 ± 20, respectively. The study of attachment and proliferation of MC3T3-E1 pre-osteoblasts using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay through indirect and direct contact methods indicated a continuous increase in metabolically active live cells with time, implying non-cytotoxicity of the synthesized hydrogel. The live-dead assay showed >95% of live cells for alginate/PAMAM hydrogels, suggesting viability of the encapsulated cells. When the percentage of PAMAM cross-linker in alginate/PAMAM hydrogel was increased from 5 to 25, the percentage degradation rate decreased from 1.1 to 0.29%/day. Given that the poly(ethylene glycol) is commonly used cross-linker for hydrogel syntheses, we compared the behavior with poly(ethylene glycol). The incorporation of poly(ethylene glycol) in alginate/PAMAM hydrogel reduced the activity of MC3T3-E1 cells and their viability compared to the alginate/PAMAM hydrogels. The protonation of amino groups in alginate/PAMAM injectables under physiological conditions led to the formation of cationic hydrogels. These cationic hydrogels showed enhanced cell encapsulation and attachment ability because of electrostatic interaction with negatively charged cell surface as determined by cell adhesion and extensions from scanning electron microscope and vinculin assay and ability of in situ calcium phosphate mineralization. These observations point toward the potential use as an injectable scaffold for cell delivery and tissue engineering applications.

Surface nanotopography-induced favorable modulation of bioactivity and osteoconductive potential of anodized 3D printed Ti-6Al-4V alloy mesh structure.

The objective of the study described here is to fundamentally elucidate the biological response of 3D printed Ti-6Al-4V alloy mesh structures that were surface modified to introduce titania nanotubes with an average pore size of ∼80 nm via an electrochemical anodization process from the perspective of enhancing bioactivity. The bioactivity of the mesh structures were analyzed through immersion test in simulated body fluid, which confirmed the nucleation and growth of fine globular nanoscale apatite on the nanoporous titania-modified (anodized) mesh structure surface, and agglomerated apatite with fine flakes of apatite crystals on as-fabricated mesh structure surface, that were rich in calcium and phosphorous. The cellular activity of bioactive anodized mesh structure was explored in terms of cell-material interactions involving adhesion, proliferation, synthesis of extracellular and intracellular proteins, differentiation, and mineralization. Cells adhered with a sheet-like morphology on as-fabricated mesh structure, whereas, on anodized mesh structure, numerous filopodia-like cellular extensions interacting with nanotube pores were observed. The formation of a bioactive nanoscale apatite, cell-nanotube interactions as imaged via electron microscopy, higher expression of proteins (actin, vinculin, fibronectin, and alkaline phosphatase (ALP)), and calcium content points toward the determining role of anodized mesh structure in modulating osteoblasts functions. The unique combination of nanoporous bioactive titania and interconnected porous architecture of anodized titanium alloy mesh structure provided a multimodal roughness surface ranging from nano to micro to macroscale, which helps in attaining strong primary and secondary fixation of the implant device along with the pathway for supply of nutrients and oxygen to cells and tissue.

Design and biological functionality of a novel hybrid Ti-6Al-4V/hydrogel system for reconstruction of bone defects.

We have designed a unique injectable bioactive hydrogel comprising of alginate, gelatin, and nanocrystalline hydroxyapatite and loaded with osteoblasts, with the ability to infiltrate into three-dimensional Ti-6Al-4V scaffolds with interconnected porous architecture, fabricated by electron beam melting. A two-step crosslinking process using the EDC/NHS and CaCl2 was adopted and found to be effective in the fabrication of cell-loaded hydrogel/Ti-6Al-4V scaffold system. This hybrid Ti-6Al-4V scaffold/hydrogel system was designed for the reconstruction of bone defects, which are difficult to heal in the absence of suitable support materials. The hybrid Ti-6Al-4V/hydrogel system favourably modulated the biological functions, namely, adhesion, proliferation, cell-to-cell, and cell-material communication because of the presence of extracellular matrix-like hydrogel in the interconnected porous structure of 3D printed Ti-6Al-4V scaffold. The hydrogel was cytocompatible, which was proven through live/dead assay, the expression level of prominent proteins for cell adhesion and cytoskeleton, including 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. Furthermore, the high bone formation ability of the hydrogel was confirmed using alkaline phosphatase assay. A high equilibrium water content (~97%) in the hydrogel enables the delivery of cells and bioactive molecules, necessary for bone tissue growth. Although not studied, the presence of hydrogel in the pores of the scaffold can provide the space for the cell migration as well as vascularization through it, required for the effective exchange of nutrients. In conclusion, we underscore that the 3D-printed Ti-6Al-4V scaffold-loaded with bioactive hydrogel to treat the bone defects significantly impacted cellular functions and cell-material interaction.

Cellular response of osteoblasts to low modulus Ti-24Nb-4Zr-8Sn alloy mesh structure.

Titanium alloys (Ti-6Al-4V and Ti-6Al-7Nb) are widely used for implants, which are characterized by high elastic modulus (∼110 GPa) with (α + ß) structure and that may induce undesirable stress shielding effect and immune responses associated with the presence of toxic elements. In this regard, we have combined the attributes of a new alloy design and the concept of additive manufacturing to fabricate 3D scaffolds with an interconnected porous structure. The new alloy is a ß-type Ti-24Nb-4Zr-8Sn (Ti2448) alloy with significantly reduced modulus. In the present study, we explore the biological response of electron beam melted low modulus Ti2448 alloy porous mesh structure through the elucidation of bioactivity and osteoblast functions. The cellular activity was explored in terms of cell-to-cell communication involving proliferation, spreading, synthesis of extracellular and intracellular proteins, differentiation, and mineralization. The formation of fine apatite-like crystals on the surface during immersion test in simulated body fluid confirmed the bioactivity of the scaffold surface, which provided the favorable osteogenic microenvironment for cell-material interaction. The combination of unique surface chemistry and interconnected porous architecture provided the desired pathway for supply of nutrients and oxygen to cells and a favorable osteogenic micro-environment for incorporation (on-growth and in-growth) of osteoblasts. The proliferation and differentiation of pre-osteoblasts and their ability to form a well mineralized bone-like extracellular matrix (ECM) by secreting bone markers (ALP, calcium, etc.) over the struts of the scaffold point toward the determining role of unique surface chemistry and 3D architecture of the Ti2448 alloy mesh structure in modulating osteoblasts functions. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 859-870, 2017.

Functional response of osteoblasts in functionally gradient titanium alloy mesh arrays processed by 3D additive manufacturing.

We elucidate here the osteoblasts functions and cellular activity in 3D printed interconnected porous architecture of functionally gradient Ti-6Al-4V alloy mesh structures in terms of cell proliferation and growth, distribution of cell nuclei, synthesis of proteins (actin, vinculin, and fibronectin), and calcium deposition. Cell culture studies with pre-osteoblasts indicated that the interconnected porous architecture of functionally gradient mesh arrays was conducive to osteoblast functions. However, there were statistically significant differences in the cellular response depending on the pore size in the functionally gradient structure. The interconnected porous architecture contributed to the distribution of cells from the large pore size (G1) to the small pore size (G3), with consequent synthesis of extracellular matrix and calcium precipitation. The gradient mesh structure significantly impacted cell adhesion and influenced the proliferation stage, such that there was high distribution of cells on struts of the gradient mesh structure. Actin and vinculin showed a significant difference in normalized expression level of protein per cell, which was absent in the case of fibronectin. Osteoblasts present on mesh struts formed a confluent sheet, bridging the pores through numerous cytoplasmic extensions. The gradient mesh structure fabricated by electron beam melting was explored to obtain fundamental insights on cellular activity with respect to osteoblast functions.

Osteoblast cellular activity on low elastic modulus Ti-24Nb-4Zr-8Sn alloy.

OBJECTIVES: Low modulus ß-titanium alloys with non-toxic alloying elements are envisaged to provide good biocompatibility and alleviate the undesired stress shielding effect. The objective of this study is to fundamentally elucidate the biological response of novel high strength-low elastic modulus Ti2448 alloy through the study of bioactivity and osteoblast cell functions. METHODS: Characterization techniques such as SEM, EDX, XRD, and fluorescence microscopy were utilized to analyze the microstructure, morphology, chemical composition, and cell adhesion. The cellular activity was explored in terms of cell-to-cell communication involving proliferation, spreading, synthesis of extracellular and intracellular proteins, differentiation, and mineralization. RESULTS: The formation of fine apatite-like crystals on the surface during immersion test in simulated body fluid confirmed the bioactivity of the surface, which provided the favorable osteogenic microenvironment for cell-material interaction. The proliferation and differentiation of pre-osteoblasts and their ability to form a well mineralized bone-like extracellular matrix (ECM) by secreting bone markers (ALP, calcium, etc.) over the surface point toward the determining role of unique surface chemistry and surface properties of the Ti-24Nb-4Zr-8Sn (Ti2448) alloy in modulating osteoblasts functions. SIGNIFICANCE: These results demonstrated that the low modulus (∼49GPa) Ti2448 alloy with non-toxic alloying elements can be used as a potential dental or orthopedic load-bearing implant material.

Understanding the response of pulsed electric field on osteoblast functions in three-dimensional mesh structures.

The endogenous electric field plays a determining role in impacting biological functions including communication with the physiological system, brain, and bone regeneration by influencing cellular functions. From this perspective, the objective of the study described here is to elucidate the effect of external electric field under dynamic conditions, in providing a guiding cue to osteoblasts in terms of cell-cell interactions and synthesis of prominent adhesion and cytoskeleton proteins. This was accomplished using pulsed direct current electric field of strength 0.1-1 V/cm. The electric field provided guided cue to the cells to migrate toward cathode. Membrane blebbing or necrosis was nearly absent in the vicinity of cathode at 0.1 and 0.5 V/cm electric field strength. Moreover, a higher cell proliferation as well as higher expression of vinculin and densely packed actin stress fibers was observed. At anode, the cells though healthy but expression of actin and vinculin was less. We underscore for the first time that the biological functionality can be favorably modulated on 3D printed scaffolds in the presence of electric field and under dynamic conditions with consequent positive effect on cell proliferation, growth, and expression level of prominent proteins.

Biological functionality and mechanistic contribution of extracellular matrix-ornamented three dimensional Ti-6Al-4V mesh scaffolds.

The 3D printed metallic implants are considered bioinert in nature because of the absence of bioactive molecules. Thus, surface modification of bioinert materials is expected to favorably promote osteoblast functions and differentiation. In this context, the objective of this study is to fundamentally elucidate the effect of cell-derived decellularized extracellular matrix (dECM) ornamented 3D printed Ti-6Al-4V scaffolds on biological functions, involving cell adhesion, proliferation, and synthesis of vinculin and actin proteins. To mimic the natural ECM environment, the mineralized ECM of osteoblasts was deposited on the Ti-6Al-4V porous scaffolds, fabricated by electron beam melting (EBM) method. The process comprised of osteoblast proliferation, differentiation, and freeze-thaw cycles to obtain decellularized extra cellular matrix (dECM), in vitro. The dECM provided a natural environment to restore the natural cell functionality of osteoblasts that were cultured on dECM ornamented Ti-6Al-4V scaffolds. In comparison to the bare Ti-6Al-4V scaffolds, a higher cell functionality such as cell adhesion, proliferation, and growth including cell-cell and cell-material interaction were observed on dECM ornamented Ti-6Al-4V scaffolds, which were characterized by using markers for focal adhesion and cytoskeleton such as vinculin and actin. Moreover, electron microscopy also indicated higher cell-material interaction and enhanced proliferation of cells on dECM ornamented Ti-6Al-4V scaffolds, supported by MTT assay. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2751-2763, 2016.

The functional response of bioactive titania-modified three-dimensional Ti-6Al-4V mesh structure toward providing a favorable pathway for intercellular communication and osteoincorporation.

The objective of the study is to fundamentally elucidate the biological response of 3D printed mesh structures subjected to plasma electrolytic oxidation process through the study of osteoblast functions. The cellular activity of plasma electrolytic-oxidized mesh structure was explored in terms of cell-to-cell communication involving proliferation, synthesis of extracellular and intracellular proteins, and mineralization. Upon plasma electrolytic oxidation of the mesh structure, a thin layer of bioactive titania with pore size 1-3 µm was nucleated on the surface. The combination of microporous bioactive titania and interconnected porous architecture provided the desired pathway for supply of nutrients and oxygen to cells and tissue and a favorable osteogenic microenvironment for tissue on-growth and in-growth, in relation to the unmodified mesh structure. The formation of a confluent layer as envisaged via electron microscopy and quantitative assessment of the expression level of proteins (actin, vinculin, and fibronectin) point toward the determining role of surface-modified mesh structure in modulating osteoblasts functions. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2488-2501, 2016.

The functional response of alginate-gelatin-nanocrystalline cellulose injectable hydrogels toward delivery of cells and bioactive molecules.

UNLABELLED: Hybrid injectable hydrogels comprising of alginate, gelatin, and nanocrystalline cellulose (NCC) were conceived and processed through adaptation of interpenetrated network of alginate-gelatin, ionic crosslinking of alginate, and supramolecular interaction approach. The design of hybrid hydrogels was based on the hypothesis that it provides an environment that is favorable for cell proliferation, exchange of nutrients via porous structure, and are characterized by mechanical properties that closely resemble the native tissue. This aspect is important for the delivery of cells or biomolecules in bone tissue engineering. The hybrid hydrogels exhibited moderate swelling behavior on formation, and the porous structure of hydrogels as imaged via SEM was envisaged to facilitate easy migration of cells and rapid transportation of biomolecules. The hybrid hydrogels exhibited desired mechanical properties and were biocompatible as confirmed though MTT assay of fibroblasts. Interestingly, osteoblasts cultured within hydrogel using bone morphogenetic protein (BMP)-2 demonstrated the capability for encapsulation of cells and induced cell differentiation. The nanocrystalline cellulose significant impacted degradation and interaction between hydrogels and cells. STATEMENT OF SIGNIFICANCE: The study fundamentally explores a hypothesis driven novel hybrid hydrogel that provides an environment for favorable growth and proliferation of cells, exchange of nutrients and mechanical properties that closely match the native tissue.

Mechanistic contribution of electroconductive hydroxyapatite-titanium disilicide composite on the alignment and proliferation of cells.

We elucidate here the mechanistic contribution of a novel electroconductive hydroxyapatite-20 wt.% titanium disilicide (HA-TiSi2) composite system in favorably modulating osteoblast functions in relation to the monolithic HA. The higher electrical conductivity of HA-TiSi2(σDC ∼ 67.117 ± 3.57 S/m) in comparison to glass sample effectively guided the electroactive myoblast, leading to their significant alignment and proliferation. This favorable behavior is attributed to the formation of small electrochemical cells between HA and TiSi2phase, which produce a small electric field, directing the electroactive myoblast to migrate and grow in a particular direction. In contrast, no impact of TiSi2on osteoblast function was observed because of their inability to respond to small electric field. However, thein vitrobioactivity in simulated body fluid indicated the nucleation and growth of apatite crystals. Moreover, in the context of load-bearing capability, the presence of 20 wt.% TiSi2in HA led to increase in the fracture toughness by ∼100%. This study underscores the effectiveness of HA-TiSi2in favorably modulating the cellular activity, myoblast in particular.

Chitosan-gelatin-based microgel for sustained drug delivery.

We describe here the proof-of-concept of synthesizing microgels via cross-linking of chitosan and gelatin with succinimide-end polyethylene glycol (PEG). The introduction of PEG provided thermo-responsiveness to microgels, and the size of microgels significantly changed with increasing temperature. Furthermore, the microgel was biocompatible as confirmed in vitro by MTT assay with NIH 3T3 fibroblast and degradable. Chitosan-based microgels exhibited strong affinity to hydrophobic drug and prolonged release of folic acid was observed at physiological temperature with varying pH. The proposed cost-effective injectable microgels have the potential to serve as the delivery vehicle of hydrophobic bioactive molecules.

Biological functionality of extracellular matrix-ornamented three-dimensional printed hydroxyapatite scaffolds.

Three-dimensional (3D) printing is considered an ideally suitable method to fabricate patient specific implantable devices. The approach enabled to produce a porous scaffold with tailored physical, mechanical, and biological properties because of the flexibility to tune the scaffold architecture. The objective of the study described was to elucidate the determining role of cell-laid extracellular matrix (ECM) in impacting biological response. In this regard, to mimic the natural ECM environment or the attributes of the native tissue, a natural ECM analogue surface was produced on the 3D printed and sintered hydroxyapatite (HA) scaffold surface by the mineralized ECM of the osteoblast. This involved the growth of osteoblast on 3D printed scaffolds, followed by differentiation to deposit the mineralized ECM on the biomaterial surface. The cells were removed from the mineralized matrix using freeze-thaw cycles to obtain a decellularized extracellular matrix (dECM) on the biomaterial surface. Subsequently, seeding of osteoblast on dECM-ornamented HA scaffolds led to 3D growth with enhanced expression of prominent proteins, actin and vinculin. Based on preliminary observations of present study, it was underscored that HA scaffolds-ornamented with dECM provided an optimized microenvironment conducive to the growth of 3D structural tissue and favorably promoted biological functionality because of the availability of an environment that promoted cell-cell and cell-scaffold interaction. The primary advantage of dECM is that it enabled constructive remodeling and promoted the formation of tissue in lieu of less functional tissue. The study opens-up a new path for printing of 3D structures suitable to treat segmental bone defects. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1343-1351, 2016.

Interplay between self-assembled structure of bone morphogenetic protein-2 (BMP-2) and osteoblast functions in three-dimensional titanium alloy scaffolds: Stimulation of osteogenic activity.

Three-dimensional cellular scaffolds are receiving significant attention in bone tissue engineering to treat segmental bone defects. However, there are indications of lack of significant osteoinductive ability of three-dimensional cellular scaffolds. In this regard, the objective of the study is to elucidate the interplay between bone morphogenetic protein (BMP-2) and osteoblast functions on 3D mesh structures with different porosities and pore size that were fabricated by electron beam melting. Self-assembled dendritic microstructure with interconnected cellular-type morphology of BMP-2 on 3D scaffolds stimulated osteoblast functions including adhesion, proliferation, and mineralization, with prominent effect on 2-mm mesh. Furthermore, immunofluorescence studies demonstrated higher density and viability of osteoblasts on lower porosity mesh structure (2 mm) as compared to 3- and 4-mm mesh structures. Enhanced filopodia cellular extensions with extensive cell spreading was observed on BMP-2 treated mesh structures, a behavior that is attributed to the unique self-assembled structure of BMP-2 that effectively communicates with the cells. The study underscores the potential of BMP-2 in imparting osteoinductive capability to the 3D printed scaffolds.

Electric field-mediated growth of osteoblasts - the significant impact of dynamic flow of medium.

The endogenous electric field plays an important role in accomplishing various functions including communication with the brain and with different parts of the physiological system, wound healing, and cellular functions. Furthermore, the endogenous electric field can be modified using the external electric field to induce changes in cell functionality. Given that the cells grow in contact with the dynamic flow of blood and nutrients, the objective of the study is to elucidate the effect of media flow (dynamic conditions) on osteoblast functions at a pulsed DC (direct current) electric field of strength of 0.5-1 V cm(-1) and compared with the static conditions (no flow of media and in the presence of an electric field). The electric field provided a guiding cue to cells to move towards the cathode. An interesting aspect of the electric field was the migration of cells towards the cathode with the axis parallel to the direction of the electric field such that the lamellipodia was aligned. Furthermore, there was an absence of membrane blebbing or necrosis at the cathode. However, cell growth and expression of proteins (actin and vinculin) were higher than the anode. In contrast, at the anode, while the cells were healthy, the cell growth was less such that the expression of vinculin was relatively low together with less densely packed actin stress fibers. It is underscored that the biological functionality is favorably altered in the presence of an electrical field under dynamic conditions with a consequent effect on cell proliferation, growth, and expression level of prominent proteins, actin and vinculin.

Osteoblast functions in functionally graded Ti-6Al-4 V mesh structures.

We describe here the combined efforts of engineering and biological sciences as a systemic approach to fundamentally elucidate osteoblast functions in functionally graded Ti-6Al-4 V mesh structures in relation to uniform/monolithic mesh arrays. First, the interconnecting porous architecture of functionally graded mesh arrays was conducive to cellular functions including attachment, proliferation, and mineralization. The underlying reason is that the graded fabricated structure with cells seeded from the large pore size side provided a channel for efficient transfer of nutrients to other end of the structure (small pore size), leading to the generation of mineralized extracellular matrix by differentiating pre-osteoblasts. Second, a comparative and parametric study indicated that gradient mesh structure had a pronounced effect on cell adhesion and mineralization, and strongly influenced the proliferation phase. High intensity and near-uniform distribution of proteins (actin and vinculin) on struts of the gradient mesh structure (cells seeded from large pore side) implied signal transduction during cell adhesion and was responsible for superior cellular activity, in comparison to the uniform mesh structure and non-porous titanium alloy. Cells adhered to the mesh struts by forming a sheet, bridging the pores through numerous cytoplasmic extensions, in the case of porous mesh structures. Intercellular interaction in porous structures provided a pathway for cells to communicate and mature to a differentiated phenotype. Furthermore, the capability of cells to migrate through the interconnecting porous architecture on mesh structures led to colonization of the entire structure. Cells were embedded layer-by-layer in the extracellular matrix as the matrix mineralized. The outcomes of the study are expected to address challenges associated with the treatment of segmental bone defects and bone-remodeling through favorable modulation of cellular response. Moreover, the study provides a foundation for a new branch of functionally graded materials with interconnected porous architecture.

The significance of grafting collagen on polycaprolactone composite scaffolds: processing-structure-functional property relationship.

The study concerns processing-structure-functional property relationship in organic-inorganic hybrid scaffolds based on grafted collagen for bone tissue engineering. Biodegradable polyester, polycaprolactone (PCL) and nanohydroxyapatite were used to fabricate three-dimensional porous scaffolds by adopting a combination of solvent casting, particulate leaching, and polymer leaching approaches. The PCL scaffold was subsequently surface modified by chemical bonding of 1,6-hexanediamine to the ester groups of PCL to introduce free NH2 groups. The introduction of NH2 groups as active sites enabled immobilization of biocompatible macromolecule, collagen, on the aminolyzed PCL via a cross-linking agent, glutaraldehyde. The osteoblasts' functions, notably cell adhesion, proliferation, and mineralization, were favorably modulated because of the chemical interaction between Arg-Gly-Asp domains in collagen molecule and integrin receptor in the cell membrane. The study underscores the significance of grafting collagen on PCL-nHA scaffold in modulating cellular activity and biological functions expanding its current use in soft tissue engineering to hard tissue regeneration.