A Combined Computational and Experimental Analysis of PLA and PCL Hybrid Nanocomposites 3D Printed Scaffolds for Bone Regeneration.

A combined computational and experimental study of 3D-printed scaffolds made from hybrid nanocomposite materials for potential applications in bone tissue engineering is presented. Polycaprolactone (PCL) and polylactic acid (PLA), enhanced with chitosan (CS) and multiwalled carbon nanotubes (MWCNTs), were investigated in respect of their mechanical characteristics and responses in fluidic environments. A novel scaffold geometry was designed, considering the requirements of cellular proliferation and mechanical properties. Specimens with the same dimensions and porosity of 45% were studied to fully describe and understand the yielding behavior. Mechanical testing indicated higher apparent moduli in the PLA-based scaffolds, while compressive strength decreased with CS/MWCNTs reinforcement due to nanoscale challenges in 3D printing. Mechanical modeling revealed lower stresses in the PLA scaffolds, attributed to the molecular mass of the filler. Despite modeling challenges, adjustments improved simulation accuracy, aligning well with experimental values. Material and reinforcement choices significantly influenced responses to mechanical loads, emphasizing optimal structural robustness. Computational fluid dynamics emphasized the significance of scaffold permeability and wall shear stress in influencing bone tissue growth. For an inlet velocity of 0.1 mm/s, the permeability value was estimated at 4.41 × 10-9 m2, which is in the acceptable range close to human natural bone permeability. The average wall shear stress (WSS) value that indicates the mechanical stimuli produced by cells was calculated to be 2.48 mPa, which is within the range of the reported literature values for promoting a higher proliferation rate and improving osteogenic differentiation. Overall, a holistic approach was utilized to achieve a delicate balance between structural robustness and optimal fluidic conditions, in order to enhance the overall performance of scaffolds in tissue engineering applications.

Adhesion strength and anti-tumor agents regulate vinculin of breast cancer cells.

The onset and progression of cancer are strongly associated with the dissipation of adhesion forces between cancer cells, thus facilitating their incessant attachment and detachment from the extracellular matrix (ECM) to move toward metastasis. During this process, cancer cells undergo mechanical stresses and respond to these stresses with membrane deformation while inducing protrusions to invade the surrounding tissues. Cellular response to mechanical forces is inherently related to the reorganization of the cytoskeleton, the dissipation of cell-cell junctions, and the adhesion to the surrounding ECM. Moreover, the role of focal adhesion proteins, and particularly the role of vinculin in cell attachment and detachment during migration, is critical, indicating the tight cell-ECM junctions, which favor or inhibit the metastatic cascade. The biomechanical analysis of these sequences of events may elucidate the tumor progression and the potential of cancer cells for migration and metastasis. In this work, we focused on the evaluation of the spreading rate and the estimation of the adhesion strength between breast cancer cells and ECM prior to and post-treatment with anti-tumor agents. Specifically, different tamoxifen concentrations were used for ER+ breast cancer cells, while even concentrations of trastuzumab and pertuzumab were used for HER2+ cells. Analysis of cell stiffness indicated an increased elastic Young's modulus post-treatment in both MCF-7 and SKBR-3 cells. The results showed that the post-treatment spreading rate was significantly decreased in both types of breast cancer, suggesting a lower metastatic potential. Additionally, treated cells required greater adhesion forces to detach from the ECM, thus preventing detachment events of cancer cells from the ECM, and therefore, the probability of cell motility, migration, and metastasis was confined. Furthermore, post-detachment and post-treatment vinculin levels were increased, indicating tighter cell-ECM junctions, hence limiting the probability of cell detachment and, therefore, cell motility and migration.

Engineering Breast Cancer Cells and hUMSCs Microenvironment in 2D and 3D Scaffolds: A Mechanical Study Approach of Stem Cells in Anticancer Therapy.

Cell biomechanics plays a major role as a promising biomarker for early cancer diagnosis and prognosis. In the present study, alterations in modulus of elasticity, cell membrane roughness, and migratory potential of MCF-7 (ER+) and SKBR-3 (HER2+) cancer cells were elucidated prior to and post treatment with conditioned medium from human umbilical mesenchymal stem cells (hUMSCs-CM) during static and dynamic cell culture. Moreover, the therapeutic potency of hUMSCs-CM on cancer cell's viability, migratory potential, and F-actin quantified intensity was addressed in 2D surfaces and 3D scaffolds. Interestingly, alterations in ER+ cancer cells showed a positive effect of treatment upon limiting cell viability, motility, and potential for migration. Moreover, increased post treatment cell stiffness indicated rigid cancer cells with confined cell movement and cytoskeletal alterations with restricted lamellipodia formation, which enhanced these results. On the contrary, the cell viability and the migratory potential were not confined post treatment with hUMSCs-CM on HER2+ cells, possibly due to their intrinsic aggressiveness. The increased post treatment cell viability and the decreased cell stiffness indicated an increased potency for cell movement. Hence, the therapy had no efficacy on HER2+ cells.

Recent Advancements in 3D Printing and Bioprinting Methods for Cardiovascular Tissue Engineering.

Recent decades have seen a plethora of regenerating new tissues in order to treat a multitude of cardiovascular diseases. Autografts, xenografts and bioengineered extracellular matrices have been employed in this endeavor. However, current limitations of xenografts and exogenous scaffolds to acquire sustainable cell viability, anti-inflammatory and non-cytotoxic effects with anti-thrombogenic properties underline the requirement for alternative bioengineered scaffolds. Herein, we sought to encompass the methods of biofabricated scaffolds via 3D printing and bioprinting, the biomaterials and bioinks recruited to create biomimicked tissues of cardiac valves and vascular networks. Experimental and computational designing approaches have also been included. Moreover, the in vivo applications of the latest studies on the treatment of cardiovascular diseases have been compiled and rigorously discussed.

Gradient 3D Printed PLA Scaffolds on Biomedical Titanium: Mechanical Evaluation and Biocompatibility.

The goal of the present investigation was to find a solution to crucial engineering aspects related to the elaboration of multi-layered tissue-biomimicking composites. 3D printing technology was used to manufacture single-layered and gradient multi-layered 3D porous scaffolds made of poly-lactic acid (PLA). The scaffolds manufacturing process was optimized after adjusting key printing parameters. The scaffolds with 60 µm side length (square-shaped pores) showed increased stiffness values comparing to the other specimens. A silicone adhesive has been further used to join biomedical titanium plates, and the PLA scaffolds; in addition, titania nanotubes (TNTs were produced on the titanium for improved adhesion. The titanium-PLA scaffold single lap joints were evaluated in micro-tensile testing. The electrochemical processing of the titanium surface resulted in a 248% increase of the ultimate strength in the overlap area for dry specimens and 40% increase for specimens immersed in simulated body fluid. Finally, the biocompatibility of the produced scaffolds was evaluated with primary cell populations obtained after isolation from bone residual tissue. The manufactured scaffolds present promising features for applications in orthopedic implantology and are worth further.

The Role of Substrate Topography and Stiffness on MSC Cells Functions: Key Material Properties for Biomimetic Bone Tissue Engineering.

The hypothesis of the present research is that by altering the substrate topography and/or stiffness to make it biomimetic, we can modulate cells behavior. Substrates with similar surface chemistry and varying stiffnesses and topographies were prepared. Bulk PCL and CNTs-reinforced PCL composites were manufactured by solvent casting method and electrospinning and further processed to obtain tunable moduli of elasticity in the range of few MPa. To ensure the same chemical profile for the substrates, a protein coating was added. Substrate topography and properties were investigated. Further on, the feedback of Wharton's Jelly Umbilical Cord Mesenchymal Stem Cells to substrates characteristics was investigated. Solvent casting scaffolds displayed superior mechanical properties compared to the corresponding electrospun films. However, the biomimetic fibrous texture of the electrospun substrates induced improved feedback of the cells with respect to their viability and proliferation. Cells' adhesion and differentiation was remarkably pronounced on solvent casting substrates compared to the electrospun substrates. Soft substates improved cells multiplication and migration, while stiff substrates induced differentiation into bone cells. Aspects related to the key factors and the ideal properties of substrates and microenvironments were clarified, aiming towards the deep understanding of the required optimum biomimetic features of biomaterials.

Combined Optimized Effect of a Highly Self-Organized Nanosubstrate and an Electric Field on Osteoblast Bone Cells Activity.

The effect of an electric field within specific intensity limits on the activity of human cells has been previously investigated. However, there are a considerable number of factors that influence the in vitro development of cell populations. In biocompatibility studies, the nature of the substrate and its topography are decisive in osteoblasts bone cells development. Further on, electrical field stimulation may activate biochemical paths that contribute to a faster, more effective self-adjustment and proliferation of specific cell types on various nanosubstrates. Within the present research, an electrical stimulation device has been manufactured and optimum values of parameters that led to enhanced osteoblasts activity, with respect to the alkaline phosphatase and total protein levels, have been found. Homogeneous electric field distribution induced by a highly organized titanium dioxide nanotubes substrate had an optimum effect on cell response. Specific substrate topography in combination with appropriate electrical stimulation enhanced osteoblasts bone cells capacity to self-adjust the levels of their specific biomarkers. The findings are of importance in the future design and development of new advanced orthopaedic materials for hard tissue replacement.

Apolipoprotein A-1 regulates osteoblast and lipoblast precursor cells in mice.

Imbalances in lipid metabolism affect bone homeostasis, altering bone mass and quality. A link between bone mass and high-density lipoprotein (HDL) has been proposed. Indeed, it has been recently shown that absence of the HDL receptor scavenger receptor class B type I (SR-B1) causes dense bone mediated by increased adrenocorticotropic hormone (ACTH). In the present study we aimed at further expanding the current knowledge as regards the fascinating bone-HDL connection studying bone turnover in apoA-1-deficient mice. Interestingly, we found that bone mass was greatly reduced in the apoA-1-deficient mice compared with their wild-type counterparts. More specifically, static and dynamic histomorphometry showed that the reduced bone mass in apoA-1(-/-) mice reflect decreased bone formation. Biochemical composition and biomechanical properties of ApoA-1(-/-) femora were significantly impaired. Mesenchymal stem cell (MSC) differentiation from the apoA-1(-/-) mice showed reduced osteoblasts, and increased adipocytes, relative to wild type, in identical differentiation conditions. This suggests a shift in MSC subtypes toward adipocyte precursors, a result that is in line with our finding of increased bone marrow adiposity in apoA-1(-/-) mouse femora. Notably, osteoclast differentiation in vitro and osteoclast surface in vivo were unaffected in the knock-out mice. In whole bone marrow, PPARγ was greatly increased, consistent with increased adipocytes and committed precursors. Further, in the apoA-1(-/-) mice marrow, CXCL12 and ANXA2 levels were significantly decreased, whereas CXCR4 were increased, consistent with reduced signaling in a pathway that supports MSC homing and osteoblast generation. In keeping, in the apoA-1(-/-) animals the osteoblast-related factors Runx2, osterix, and Col1a1 were also decreased. The apoA-1(-/-) phenotype also included augmented CEPBa levels, suggesting complex changes in growth and differentiation that deserve further investigation. We conclude that the apoA-1 deficiency generates changes in the bone cell precursor population that increase adipoblast, and decrease osteoblast production resulting in reduced bone mass and impaired bone quality in mice.

MWCNTs enhance hBMSCs spreading but delay their proliferation in the direction of differentiation acceleration.

Investigating the ability of films of pristine multiwalled nanotubes (MWCNTs) to influence human mesenchymal stem cells' proliferation, morphology, and differentiation into osteoblasts, we concluded to the following: A. MWCNTs delay the proliferation of hBMS cells but increase their differentiation. The enhancement of the differentiation markers could be a result of decreased proliferation and maturation of the extracellular matrix B. Cell spread on MWCNTs toward a polygonal shape with many thin filopodia to attach to the surfaces. Spreading may be critical in supporting osteogenic differentiation in pre-osteoblastic progenitors, being related with cytoskeletal tension. C. hBMS cells prefer MWCNTs than tissue plastic to attach and grow, being non-toxic to these cells. MWCNTs can be regarded as osteoinductive biomaterial topographies for bone regenerative engineering.

Multiwalled carbon nanotubes enhance human bone marrow mesenchymal stem cells' spreading but delay their proliferation in the direction of differentiation acceleration.

Investigating the ability of films of pristine (purified, without any functionalization) multiwalled carbon nanotubes (MWCNTs) to influence human bone marrow mesenchymal stem cells' (hBMSCs) proliferation, morphology, and differentiation into osteoblasts, we concluded to the following: A. MWCNTs delay the proliferation of hBMSCs but increase their differentiation. The enhancement of the differentiation markers could be a result of decreased proliferation and maturation of the extracellular matrix B. Cell spread on MWCNTs toward a polygonal shape with many thin filopodia to attach to the surfaces. Spreading may be critical in supporting osteogenic differentiation in pre-osteoblastic progenitors, being related with cytoskeletal tension. C. hBMSCs prefer MWCNTs than tissue plastic to attach and grow, being non-toxic to these cells. MWCNTs can be regarded as osteoinductive biomaterial topographies for bone regenerative engineering.

Cellular function and adhesion mechanisms of human bone marrow mesenchymal stem cells on multi-walled carbon nanotubes.

Multiwalled carbon nanotubes (MWCNTs) are considered to be excellent reinforcements for biorelated applications, but, before being incorporated into biomedical devices, their biocompatibility need to be investigated thoroughly. We investigated the ability of films of pristine MWCNTs to influence human mesenchymal stem cells' proliferation, morphology, and differentiation into osteoblasts. Moreover, the selective integrin subunit expression and the adhesion mechanism to the substrate were evaluated on the basis of adherent cell number and adhesion strength, following the treatment of cells with blocking antibodies to a series of integrin subunits. Results indicated that MWCNTs accelerated cell differentiation to a higher extent than tissue culture plastic, even in the absence of additional biochemical inducing agents. The pre-treatment with anti-integrin antibodies decreased number of adherent cells and adhesion strength at 4-60%, depending on integrin subunit. These findings suggest that pristine MWCNTs represent a suitable reinforcement for bone tissue engineering scaffolds.

Detachment strength of human osteoblasts cultured on hydroxyapatite with various surface roughness. Contribution of integrin subunits.

Hydroxyapatite (HA) has been widely used as a bone substitute in dental, maxillofacial and orthopaedic surgery and as osteoconductive bone substitute or precoating of pedicle screws and cages in spine surgery. The aim of the present study was to investigate the osteoblastic adhesion strength on HA substrata with different surface topography and biochemistry (pre-adsorption of fibronectin) after blocking of specific integrin subunits with monoclonal antibodies. Stoichiometric HA was prepared by precipitation followed by ageing and characterized by SEM, EDX, powder XRD, Raman spectroscopy, TGA, and specific surface area analysis. Human bone marrow derived osteoblasts were cultured on HA disc-shaped substrata which were sintered and polished resulting in two surface roughness grades. For attachment evaluation, cells were incubated with monoclonal antibodies and seeded for 2 h on the substrata. Cell detachment strength was determined using a rotating disc device. Cell detachment strength was surface roughness, fibronectin preadsorption and intergin subunit sensitive.

On the biocompatibility between TiO2 nanotubes layer and human osteoblasts.

Titanium and its alloys are the most popular biomaterials replacing hard tissues in implant surgeries. Clinicians are generally pleased by titanium mechanical properties and non-toxicity performances; on the other hand, there have been reported several cases of titanium implantation failure, phenomenon explained sometimes as "non adherence of human tissue to the metallic surface." Yet, researchers reported that titanium surfaces are favorable for osteoblasts adhesion. Therefore, titanium integration into the human body remains an unsolved problem. In the present study, biocompatibility tests were performed on titanium and TiO(2) nanotubes substrates, involving human bone marrow cells. The combination of a newly developed analytical model based on the hybrid interphase concept, applicable to systems consisting of inert materials when in contact with living tissues, together with experimental results, confirmed previous research studies and lead to the conclusion that osteoblasts adhere efficiently to titanium surfaces. However, the present results suggest that osteoblasts strong anchorage at the very first moment of their contact with the metallic material leads to their apoptosis. It is most probable that in several cases this is the reason of failed implantation surgeries involving titanium.

Biomechanical and structural changes following the decellularization of bovine pericardial tissues for use as a tissue engineering scaffold.

To achieve natural scaffolds for tissue engineering applications we decellularized bovine pericardial (BP) tissues according to two different protocols: a novel treatment based on Triton(®) X-100 (12 h, 4 °C) (BP1) and a trypsin/EDTA treatment (37 °C, 48 h) (BP2). Results were compared with commercially available acellular xenogeneic biomaterials, Veritas(®) and Collamed(®). Biomechanical characteristics, high (E(h)) and low (E(l)) modulus of elasticity, of the fresh untreated tissue varied with the anatomical direction (apex to base (T) to transverse (L)) (mean ± SDEV): (41.63 ± 14.65-48.12 ± 10.19 MPa and 0.27 ± 0.05-0.30 ± 0.12 MPa respectively). BP1 had no mechanical effect (44.65 ± 19.73-52.67 ± 7.59 MPa and 0.37 ± 0.14-0.37 ± 0.11 MPa, respectively) but BP2 resulted in significant decrease in E(h) and E(l) (20.96 ± 8.17-36.82 ± 3.23 MPa and 0.20 ± 0.06-0.23 ± 0.06 MPa). Hysteresis ratio (h) varied (19-26 % of the loading energy) independently of anatomical direction. Glycosaminoglycans content was unaffected by BP1, while 22 % of chondroitin/dermatan sulphate and 60 % of hyaluronan were removed after BP2 treatment. Endothelial cell adhesion was achieved after 24 h and 3 days cell culture.

Effects of physiological mechanical strains on the release of growth factors and the expression of differentiation marker genes in human osteoblasts growing on Ti-6Al-4V.

Mechanical loading factors at the bone-implant interface are critical for the osseointegration and clinical success of the implant. The aim of the present investigation was to study the effects of mechanical strain on the orthopedic biomaterial Ti-6Al-4V/osteoblast interface, using an in vitro model. Homogeneous strain was applied to human bone marrow derived osteoblasts (HBMDOs) cultured on Ti-6Al-4V, at physiological levels (strain magnitudes 500 microstrain (microepsilon) and 1000 microepsilon, at frequencies of load application 0.5 Hz and 1 Hz), by a mechanostimulatory system, based on the principle of four-point bending. Semi-quantitative reverse transcription-polymerase chain reaction (sqRT-PCR) was used to determine the mRNA expression of Cbfa1 and osteocalcin at different loading conditions. The release of growth factors as a response to stretch was also investigated by transferring stretch-conditioned media to nonstretched cells and by measuring their effect on the regulation of DNA synthesis. Mechanical loading was found to contribute to the regulation of osteoblast differentiation by influencing the level of the osteoblast-specific transcription factor Cbfa1, both at the mRNA and protein level, and also the level of osteocalcin, which is regarded as the most osteoblast-specific gene. Both genes were differentially expressed shortly after the application of different mechanical stimuli, in terms of strain frequency, magnitude, and time interval. Media conditioned from mechanically stressed HBMDOs stimulate DNA synthesis more intensely compared to media conditioned from unstressed control cultures, indicating that mechanical strain induces the release of a mitogenic potential that regulates cell proliferation.

Estimation of hydrodynamic shear stresses developed on human osteoblasts cultured on Ti-6Al-4V and strained by four point bending. Effects of mechanical loading to specific gene expression.

The aim of the present investigation was to study the effects of mechanical strain on the orthopedic biomaterial Ti-6Al-4V-osteoblast interface, using an in vitro model. Homogeneous strain was applied to Human Bone Marrow derived Osteoblasts (HBMDOs) cultured on Ti-6Al-4V, at levels which are considered physiological, by a four-point bending mechanostimulatory system. A simple model for the estimation of maximum hydrodynamic shear stresses developed on cell culture layer and induced by nutrient medium flow during mechanical loading, as a function of the geometry of the culture plate and the load characteristics, is proposed. Shear stresses were lower than those which can elicit cell response. Mechanical loading was found that contributes to the regulation of osteoblast differentiation by influencing the expression of the osteoblast-specific transcription factor Cbfa1, both at the mRNA and protein level, and also the osteocalcin expression, whereas osteopontin gene expression was unaffected by mechanical loading at all experimental conditions.

Biomechanical comparison of callus over a locked intramedullary nail in various segmental bone defects in a sheep model.

BACKGROUND: Little has been written about the size of a bone defect that can be restored with one-stage lengthening over a reamed intramedullary nail. MATERIAL/METHODS: Sixteen adult female sheep were divided into four main groups: a simple osteotomy group (group I) and three segmental defect groups (1-, 2-, and 3-cm gaps, groups II-IV). One intact left tibia from each group was also used as the non-osteotomized intact control group (group V). In all cases, the osteotomy was fixed with an interlocked Universal Humeral Nail after reaming to 7 mm. Healing of the osteotomies was evaluated after 16 weeks by biomechanical testing. The examined parameters were torsional stiffness, shear stress, and angle of torsion at the time of fracture. RESULTS: The regenerate bone obvious in x-rays in the groups with 1- and 2-cm gaps had considerable mechanical properties. Torsional stiffness in these two groups was nearly equal and its value was about 60% of the stiffness of the simple osteotomy group. Gradually decreasing stiffness was observed as the osteotomy gap increased. No significant differences were found among the angles of torsion at fracture for the various osteotomies or the intact bone. These results showed that the group with 1-cm gap had 65% of the shear stress at failure of the simple osteotomy group. CONCLUSIONS: The authors believe that there is evidence indicating that intramedullary nailing could be a reasonable option when one-stage lengthening of a long bone by 1 or 2 cm is contemplated.

Biomechanical evaluation of conventional internal contemporary spinal fixation techniques used for stabilization of complete sacroiliac joint separation: a 3-dimensional unilaterally isolated experimental stiffness study.

STUDY DESIGN: Comparative 3-dimensional biomechanical testing. OBJECTIVE: To compare 5 fixation techniques, 3 using screws or screw and plates and 2 spinal, used for stabilization of complete unilateral sacroiliac dislocation in composite models. SUMMARY OF BACKGROUND DATA: Harrington compression rods have been used for posterior iliosacral stabilization. Recently, the use of compact spinal instrumentation has been introduced for stabilization of iliosacral joint separation to achieve immediate and permanent stability, allowing early mobilization. To the authors' knowledge, no comparative mechanical studies between commonly used internal fixation techniques and contemporary spinal instrumentation have been performed. METHODS: Fifteen identical composite models of the left hemipelvis and sacrum were used to simulate consistently the "worst-case scenario" of complete unilateral sacroiliac dislocation. Subgroups of 3 models each were used to apply 5 (A-E) alternative fixation iliosacral joint fixation techniques: 1 multiaxial 7.5 mm Cotrel-Dubousset screw inserted in the posterior superior iliac spine and connected with a long Cotrel-Dubousset horizontal rod with 6.5 mm multiaxial Cotrel-Dubousset screws inserted bilaterally in the S1 pedicles (technique A); 1 multiaxial 7.5 mm Cotrel-Dubousset titanium pedicle screw inserted in the posterior superior iliac spine and connected with a short horizontal Cotrel-Dubousset-rod to a 6.5 mm multiaxial Cotrel-Dubousset-screw inserted to the ipsilateral S1 pedicle (technique B); 1, 6.5 mm cancellous AO-screw (technique C); 2, 6.5 mm cancellous AO screws (technique D); and 2 dynamic stainless steel compression plates (technique E) placed anteriorly. Constructs were biomechanically tested. The ilium was unilaterally rigidly fixed, the sacrum was put horizontal in the mediolateral direction with a forward tilt of 30 degrees (close to physiologic conditions) in the sagittal plane, and a vertical quasi-static compressive load ranging from 0 to 500 N was applied on the endplate of S1, reproducing a "worst case" loading scenario. Construct stiffness, frontal plus sagittal kinematics, and iliosacral joint gap size for all 5 techniques were measured. RESULTS: The construct stiffness (N/mm +/- standard deviation) ranged for model: A, 121 +/- 18; B, 78 +/- 10; C, 168 +/- 13; D, 193 +/- 42; and E, 145 +/- 4. All other parameters exhibited minor variations between the different techniques of fixation: at the 400 N load level, the maximum iliosacral gap globally ranged 0.9-2.8 mm, the maximum mediolateral sacral tilt ranged 1.3-2.4 degrees, and the maximum anteroposterior sacral tilt ranged 0.6-3.0 degrees. CONCLUSIONS: The iliosacral fixation with 2 6.5 mm AO-cancellous screws for complete sacroiliac dislocation demonstrated the highest stiffness and the short spinal instrumentation the poorest stiffness. All other fixation techniques could be generally considered of equivalent stability value.

Experimental usage of hydroxyapatite preadsorption with fibronectin to increase permanent stability and longevity of spinal implants.

UNLABELLED: Hydroxyapatite has been used in orthopaedic and particularly in spinal surgery by precoating implants to indirectly increase osteoblasts' adhesion and subsequently their stability and longevity. Fibronectin preadsorption synergistically with appropriately constructed hydroxyapatite's surface texture to enhance osteoblasts' adhesion has not been, to the authors' knowledge, previously investigated. In osteoporotic spines, methods to increase implant stability (pedicle screws and cages) are of major value. OBJECTIVE: This experimental study investigated the contribution of fibronectin preadsorption to enhance osteoblasts' adhesion and strength on hydroxyapatite. METHODS: Hydroxyapatite substrata with two different surface roughnesses (rough HA180 and the smooth HA1200) were produced and human osteoblasts were seeded on them after culture. Prior to osteoblasts seeding, the hydroxyapatite substrata were immersed in fibronectin solution. Osteoblast attachment on each of the two hydroxyapatite substrata was evaluated by recording the number of cells, while the osteoblast's adhesion strength was determined by measuring the shear stress required to detach the cells from the hydroxyapatite substrates. RESULTS: Fibronectin preadsorption increased the number of attached osteoblasts on smooth and rough hydroxyapatite substratum at 40% and 62% respectively, while it increased osteoblast attachment strength on the smooth and rough substratum at 165% and 73% respectively. CONCLUSIONS: Fibronectin preadsorption and smooth hydroxyapatite surface texture synergistically increased the adhesion's strength of human osteoblasts "in vitro", while preadsorption and rough hydroxyapatite surface increased the number of attached osteoblasts. Further studies in primates and human beings should be carried out to disclose the clinical relevance of the above mentioned observations in spine surgery.

Fibronectin preadsorbed on hydroxyapatite together with rough surface structure increases osteoblasts' adhesion "in vitro": the theoretical usefulness of fibronectin preadsorption on hydroxyapatite to increase permanent stability and longevity in spine implants.

OBJECTIVE: The aim of this study was to investigate the contribution of fibronectin (FN) preadsorption to enhance osteoblast adhesion and strength on hydroxyapatite (HA) used either as osteoconductive bone substitute or precoating of pedicle screws and cages in spine surgery. METHODS: HA substrata with two different surface roughness values (rough HA180 and smooth HA1200) were produced, and human osteoblasts were seeded after culturing on them. Prior to osteoblast seeding, the HA substrata were immersed in FN solution. Osteoblast attachment on each of the two HA substrata was evaluated by recording the number of cells, whereas osteoblast adhesion strength was determined by measuring the shear stress required to detach the cells from the HA substrates. RESULTS: FN preadsorption increased the number of attached osteoblasts on smooth and rough HA substratum at 40% and 62%, respectively, whereas it increased osteoblast attachment strength on the smooth and rough substratum at 165% and 73%, respectively. CONCLUSIONS: This study showed that FN preadsorption and rough HA surface texture synergistically increased "in vitro" both the number and the adhesion strength of human osteoblasts. Further studies in primates and humans should be carried out to disclose the clinical relevance (increase implant's stability and longevity) of the above-mentioned observations.