Nano-hydroxyapatite Scaffolds Research Articles

The Impact of Nano-Hydroxyapatite Scaffold Enrichment on Bone Regeneration In Vivo-A Systematic Review.

In order to ensure improved and accelerated bone regeneration, nano-hydroxyapatite scaffolds are often enriched with different bioactive components to further accelerate and improve bone healing. In this review, we critically examined whether the enrichment of nHAp/polymer scaffolds with growth factors, hormones, polypeptides, microRNAs and exosomes improved new bone formation in vivo. Out of 2989 articles obtained from the literature search, 106 papers were read in full, and only 12 articles met the inclusion criteria for this review. Several bioactive components were reported to stimulate accelerated bone regeneration in a variety of bone defect models, showing better results than bone grafting with nHAp scaffolds alone. The results indicated that composite materials based on nHAp are excellent candidates as bone substitutes, while nHAp scaffold enrichment further accelerates bone regeneration. The standardization of animal models should be provided in order to clearly define the most significant parameters of in vivo studies. Only in this way can the adequate comparison of findings from different in vivo studies be possible, further advancing our knowledge on bone regeneration and enabling its translation to clinical settings.

Biomimetic highly porous nanocellulose–nanohydroxyapatite scaffolds for bone tissue engineering

Wood-derived TEMPO-oxidized cellulose nanofibrils (CNFs) have potential as scaffolding for bone tissue engineering. Although biocompatible, the material lacks osteoconductive and appropriate mechanical properties. Incorporation of nano-hydroxyapatite (nHA) and modification of scaffold preparation methods could improve applicability. In this study, freeze-dried porous scaffolds were prepared using a range of nHA (0, 20, 33, 50%) and CNF compositions. Not only the microarchitecture but also the chemical composition of the scaffolds was studied. Osteoblast-like osteosarcoma derived cells (Saos-2) were cultured on the scaffolds and their responses (viability, attachment, proliferation, and osteogenic phenotype) to the different scaffolds were documented. The results show that incorporation of nHA influenced the microarchitecture, mechanical stiffness and surface properties of the scaffolds. Moreover, biological characterization demonstrated good cell viability in all the groups. However, the increase of nHA concentration beyond 20% does not offer further advantages. It is concluded that the incorporation of 20% nHA resulted in the widest and most biomimetic pore size distribution, increased surface roughness and improved protein adsorption. These changes in material properties enhanced cell spreading and the osteogenic gene expression of osteoblast-like cells seeded on the scaffolds. Moreover, 20% nHA warrants further investigation as a potential scaffolding material for bone tissue engineering.Graphical abstract

Preparation and properties of a 3D printed nHA/PLA bone tissue engineering scaffold loaded with a β-CD-CHX combined dECM hydrogel.

Jaw defects, which can result from a multitude of causes, significantly affect the physical well-being and psychological health of patients. The repair of these infected defects presents a formidable challenge in the clinical and research fields, owing to their intricate and diverse nature. This study aims to develop a personalized bone tissue engineering scaffold that synergistically offers antibacterial and osteogenic properties for treating infected maxillary defects. This study engineered a novel temperature-sensitive, sustained-release hydrogel by amalgamating β-cyclodextrin (β-CD) with chlorhexidine (CHX) and a decellularized extracellular matrix (dECM). This hydrogel was further integrated with a polylactic acid (PLA)-nano hydroxyapatite (nHA) scaffold, fabricated through 3D printing, to form a multifaceted composite scaffold (nHA/PLA/dECM/β-CD-CHX). Drug release assays revealed that this composite scaffold ensures prolonged and sustained release. Bacteriological studies confirmed that the β-CD-CHX loaded scaffold exhibits persistent antibacterial efficacy, thus effectively inhibiting bacterial growth. Moreover, the scaffold demonstrated robust mechanical strength. Cellular assays validated its superior biocompatibility, attributed to dECM and nHA components, significantly enhancing the proliferation, adhesion, and osteogenic differentiation of osteogenic precursor cells (MC3T3-E1). Consequently, the nHA/PLA/dECM/β-CD-CHX composite scaffold, synthesized via 3D printing technology, shows promise in inducing bone regeneration, preventing infection, and facilitating the repair of jaw defects, positioning itself as a potential breakthrough in bone tissue engineering.

Polycaprolactone, polylactic acid, and nanohydroxyapatite scaffolds obtained by electrospinning and 3D printing for tissue engineering

There is a deficit for bone tissue natural grafts that seek to be covered with synthetic substitutes. Scaffolds generated with 3D printing and electrospinning allow adequate mechanical properties maintaining a structure appropriate for cell growth. Here, a scaffold made up of three-dimensional (3D) printed PLA frameworks added with PCL/PLA/nHA nanofibers was manufactured. The framework showed mechanical properties similar to other reported bone substitutes, while the nanofibers showed diameters between 200 and 850 nm. Scaffolds were suitable for cell adhesion and proliferation when evaluated with fibroblasts, showing cell proliferation into the nanofiber network, a fundamental aspect in tissue engineering.

Viability of Collagen Matrix Grafts Associated with Nanohydroxyapatite and Elastin in Bone Repair in the Experimental Condition of Ovariectomy.

Bone lesions have the capacity for regeneration under normal conditions of the bone metabolism process. However, due to the increasing incidence of major traumas and diseases that cause bone-mineral deficiency, such as osteoporosis, scaffolds are needed that can assist in the bone regeneration process. Currently, natural polymeric scaffolds and bioactive nanoparticles stand out. Therefore, the objective of the study was to evaluate the osteoregenerative potential in tibiae of healthy and ovariectomized rats using mineralized collagen and nanohydroxyapatite (nHA) scaffolds associated with elastin. The in-vivo experimental study was performed with 60 20-week-old Wistar rats, distributed into non-ovariectomized (NO) and ovariectomized (O) groups, as follows: Controls (G1-NO-C and G4-O-C); Collagen with nHA scaffold (G2-NO-MSH and G5-O-MSH); and Collagen with nHA and elastin scaffold (G3-NO-MSHC and G6-O-MSHC). The animals were euthanized 6 weeks after surgery and the samples were analyzed by macroscopy, radiology, and histomorphometry. ANOVA and Tukey tests were performed with a 95% CI and a significance index of p < 0.05. In the histological analyses, it was possible to observe new bone formed with an organized and compact morphology that was rich in osteocytes and with maturity characteristics. This is compatible with osteoconductivity in both matrices (MSH and MSHC) in rats with normal conditions of bone metabolism and with gonadal deficiency. Furthermore, they demonstrated superior osteogenic potential when compared to control groups. There was no significant difference in the rate of new bone formation between the scaffolds. Ovariectomy did not exacerbate the immune response but negatively influenced the bone-defect repair process.

Investigation into relationships between design parameters and mechanical properties of 3D printed PCL/nHAp bone scaffolds.

Scaffolds are of great importance in tissue engineering applications as they provide a mechanically supportive environment for cellular activity, which is particularly necessary for hard tissues such as bone. Notably, the mechanical properties of a scaffold vary with differing design parameters such as those related to scaffold height and internal structure. Thus, the present study aimed to explore the relationship between design parameters and mechanical properties of composite polycaprolactone (PCL) and nano-hydroxyapatite (nHAp) scaffolds fabricated by three-dimensional (3D) printing. We designed and printed scaffolds with different internal structures (lattice and staggered) and varying heights (4, 6, 8 and 10 layers), and consistent porosity (50%) for the purpose of comparison. Then, we examined the scaffold microstructure (pore size and penetration between layers) using scanning electron microscopy (SEM) and mechanical properties (elastic modulus and yield strength) using compressive testing. Our results illustrated that the microstructural parameters were related to scaffold design. At higher heights, pore size increased while penetration between layers decreased; thus, mechanical properties were affected. Results of mechanical testing demonstrated that for lattice scaffolds, elastic modulus was similar for 6 vs 4, and 8 vs 4 layers but ~33% lower for 10 layers vs 4 layers. Similarly, yield strength was comparable for 6 vs 4, and 8 vs 4 layers but ~27% lower for 10 layers vs 4 layers. With staggered scaffolds, when compared to 4-layer results, elastic modulus was similar for 6 layers but was ~43% lower for 8 layers and ~38% lower for 10 layers. Staggered scaffolds had ~38%, ~51%, and ~76% lower yield strength when the number of layers were increased from 4 to 6, 8, and 10 layers, respectively. When comparing lattice and staggered scaffolds with the same layer number, elastic modulus was similar, apart from 8-layer scaffolds where the staggered design was ~42% lower than lattice. Yield strength was similar between 4-layer staggered and lattice scaffolds, while staggered scaffolds with 6, 8, and 10 number of layers showed ~43%, ~45%, ~68% lower strength, respectively, than those found in lattice scaffolds with the same layer numbers. Mechanical properties of 3D printed scaffolds depended on scaffold height for both lattice and staggered internal structures. Staggered scaffolds had lower mechanical properties than the lattice scaffolds with the same height and were more sensitive to the change in scaffold height. Taken together, lattice scaffolds demonstrated the advantages of more stable mechanical properties over staggered scaffolds. Also, scaffolds with lower height were more promising in terms of mechanical properties compared to scaffolds with greater height.

Investigation into Biocompatibility and In Vitro Osteogenic Differentiation Ability of New Degradable Polymer Material (Poly-glycerolsebacate-maleic Anhydride) Combined with Nano-Hydroxyapatite Scaffolds

This work explored the biocompatibility and In Vitro osteogenic differentiation ability of new degradable polymer scaffold to provide some references for the research focusing on applying the scaffold for osseous tissue engineering. Polyglycerolsebacate-maleic anhydride (PGS-M) and Nano-hyproxyapatite (n-HA) were utilized to prepare a new scaffold, which was called PGS-M/n-HA herein. Besides, scanning electron microscope (SEM) was employed to observe the morphology and porosity of the PGS-M/n-HA, and its In Vitro degradation rate and mechanical property (MP) were analyzed. Human bone marrow mesenchymal stem cells (HBMSCs) (Ctrl group), osteoblast inducing conditional media (OICM group), PGS-M/n-HA group, and PGS-M/n-HA combining with OICM (OICM+PGS-M/n-HA group) were cultured. After that, the differences in cell adhesion rate, viability, and alkaline phosphatase (AKP) activity were compared and analyzed. It was demonstrated that porosities of PGS-M and PGS-M/n-HA declined while the constrained modulus increased. Based on those in the Ctrl group, the cell viability and AKP activity in the OICM group were enhanced. In contrast to those in PGS-M/n-HA group, cell adhesion rate and AKP activity were both enhanced in the OICM+PGS-M/n-HA group. Therefore, it suggested that PGS-M/n-HA scaffold possessed high degradability, MP, and biocompatibility, and could induce the differentiation of HBMSCs into osteoblasts.

Chitosan-vancomycin hydrogel incorporated bone repair scaffold based on staggered orthogonal structure: a viable dually controlled drug delivery system.

In clinical practice, challenges remain in the treatment of large infected bone defects. Bone tissue engineering scaffolds with good mechanical properties and antibiotic-controlled release are powerful strategies for infection treatment. In this study, we prepared polylactic acid (PLA)/nano-hydroxyapatite (nHA) scaffolds with vertical orthogonal and staggered orthogonal structures by applying 3D printing technology. In addition, vancomycin (Van)-based chitosan (CS) hydrogel (Gel@Van) was loaded on the scaffold (PLA/nHA/CS-Van) to form a local antibiotic release system. The microstructure of the composite scaffold had high porosity with interconnected three-dimensional networks. The mechanical properties of the PLA/nHA/CS-Van composite scaffold were enhanced by the addition of CS-Van. The results of the water contact angle analysis showed that the hydrophilicity of the drug-loaded scaffold improved. In addition, the composite scaffold could produce sustained release in vitro for more than 8 weeks without adverse effects on the proliferation and differentiation of mouse embryonic osteoblasts (MC3T3-E1), which confirmed its good biocompatibility. During the in vitro antimicrobial study, the composite scaffold effectively inhibited the growth of Staphylococcus aureus (S. aureus). Therefore, our results suggest that the PLA/nHA/CS-Van composite scaffold is a promising strategy for treating infected bone defects.

Dual-Nozzle 3D Printed Nano-Hydroxyapatite Scaffold Loaded with Vancomycin Sustained-Release Microspheres for Enhancing Bone Regeneration

Successful treatment of infectious bone defect remains a major challenge in the orthopaedic field. At present, the conventional treatment for infectious bone defects is surgical debridement and long-term systemic antibiotic use. It is necessary to develop a new strategy to achieve effective bone regeneration and local anti-infection for infectious bone defects. Firstly, vancomycin / poly (lactic acid-glycolic acid) sustained release microspheres (VAN/PLGA-MS) were prepared. Then, through the dual-nozzle 3D printing technology, VAN/PLGA-MS was uniformly loaded into the pores of nano-hydroxyapatite (n-HA) and polylactic acid (PLA) scaffolds printed in a certain proportion, and a composite scaffold (VAN/MS-PLA/n-HA) was designed, which can not only promote bone repair but also resist local infection. Finally, the performance of the composite scaffold was evaluated by in vivo and in vitro biological evaluation. The in vitro release test of microspheres showed that the release of VAN/PLGA-MS was relatively stable from the second day, and the average daily release concentration was about 15.75 μg/mL, which was higher than the minimum concentration specified in the guidelines. The bacteriostatic test in vitro showed that VAN/PLGA-MS had obvious inhibitory effect on Staphylococcus aureus ATCC-29213. Biological evaluation of VAN/MS-PLA/n-HA scaffolds in vitro showed that it can promote the proliferation of adipose stem cells. In vivo biological evaluation showed that VAN/MS-PLA/n-HA scaffold could significantly promote bone regeneration. Our research shows that VAN/MS-PLA/n-HA scaffolds have satisfying biomechanical properties, effectively inhibit the growth of Staphylococcus aureus, with good biocompatibility, and effectiveness on repairing bone defects. The VAN/MS-PLA/n-HA scaffold provide the clinic with an application prospect in bone tissue engineering.

Three-dimensional printed poly(l-lactide) copolymers with nano-hydroxyapatite scaffolds for enhanced osteogenic and regenerative activities in bone tissue engineering

Biocompatible porous scaffolds with adjustable degradation rate and mechanical properties have attracted increasing interest in tissue engineering. 3D printing technology is an effective method for the rapid preparation of bone tissue engineering scaffolds. In this study, bone conductive poly(l-lactide-ran-p-dioxanone-ran-glycolide) (PLPG)/nano-hydroxyapatite (nHAP) (P/H) composites are prepared by solution blending method, which are used to fabricate P/H scaffolds by 3D printing technology. The physicochemical properties of the P/H composites are examined by DSC, XRD, and contact angle goniometry. Furthermore, the surface morphology, mechanical properties, biodegradation ability, and in vitro biocompatibility of the P/H scaffolds are investigated. The results indicate the PLPG copolymers have a highly printability with nHAP, and the mechanical and biodegradation properties of the P/H scaffolds can be tuned by adjusting the ratio of PLPG and nHAP. The in vitro biological experiments show that the P/H scaffolds are excellent substrates for the adhesion, growth, and proliferation of MC3T3-E1.

Fabrication and characterization of bioinspired nanohydroxyapatite scaffolds with different porosities

This study aimed to fabricate and characterize bovine nanohydroxyapatite (nano-HAp) scaffolds at a low sintering temperature. The compact bone/starch systems were formed using the inorganic matrix of trabecular bone powder and achira isolated starch. To generate the macroporosity, 0, 10, 15, and 20% achira starch granules were added. To obtain nano-HAp scaffolds, the compact bone/starch systems were incinerated at 600 °C, removing organic material. The scaffolds exhibit the presence of Ca, P, Sr, Ba, Fe, Na, Mg, K, and Si came from the HAp with a slight K increase because of the mineral contribution of the starch. Infrared spectroscopy helped determine the elimination of organic materials and evidenced carbonated nano-HAp. X-ray diffraction allows to determine the presence of nano-HAp crystals. Moreover, the X-ray tomography revealed an increased macroporosity (30–38%) for the scaffolds as the starch percentage increased and scanning electron microscopy (SEM) average pore length size for HAp, HAp-A10, HAp-A15, and HAp-A20 were 356.71 ± 156.71, 548.91 ± 131.99, 620.94 ± 147.93, and 728.16 ± 160.98 μm, respectively. In the same way, SEM images of the scaffolds evidenced the presence of micro and nano porosities. The N2 adsorption-desorption analysis corroborated the presence of mesoporosity with average pore diameters of 3.66 and 3.32 nm in the samples before and after the calcining process, respectively. Thus, thermal treatments did not significantly affect the mesoporous size. In addition, effective Young's modulus is governed by the intrinsic properties of HAp, while the porosity governs the plastic and breaking point. The coalescence phenomena create the scaffold macro architecture.

Effect of high content nanohydroxyapatite composite scaffolds prepared via melt extrusion additive manufacturing on the osteogenic differentiation of human mesenchymal stromal cells.

The field of bone tissue engineering seeks to mimic the bone extracellular matrix composition, balancing the organic and inorganic components. In this regard, additive manufacturing (AM) of high content calcium phosphate (CaP)-polymer composites holds great promise towards the design of bioactive scaffolds. Yet, the biological performance of such scaffolds is still poorly characterized. In this study, melt extrusion AM (ME-AM) was used to fabricate poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT)-nanohydroxyapatite (nHA) scaffolds with up to 45wt% nHA, which presented significantly enhanced compressive mechanical properties, to evaluate their in vitro osteogenic potential as a function of nHA content. While osteogenic gene upregulation and matrix mineralization were observed on all scaffold types when cultured in osteogenic media, human mesenchymal stromal cells did not present an explicitly clear osteogenic phenotype, within the evaluated timeframe, in basic media cultures (i.e. without osteogenic factors). Yet, due to the adsorption of calcium and inorganic phosphate ions from cell culture media and simulated body fluid, the formation of a CaP layer was observed on PEOT/PBT-nHA 45wt% scaffolds, which is hypothesized to account for their bone forming ability in the long term in vitro, and osteoconductivity in vivo.

Development of a Gene-Activated Scaffold Incorporating Multifunctional Cell-Penetrating Peptides for pSDF-1α Delivery for Enhanced Angiogenesis in Tissue Engineering Applications.

Non-viral gene delivery has become a popular approach in tissue engineering, as it permits the transient delivery of a therapeutic gene, in order to stimulate tissue repair. However, the efficacy of non-viral delivery vectors remains an issue. Our lab has created gene-activated scaffolds by incorporating various non-viral delivery vectors, including the glycosaminoglycan-binding enhanced transduction (GET) peptide into collagen-based scaffolds with proven osteogenic potential. A modification to the GET peptide (FLR) by substitution of arginine residues with histidine (FLH) has been designed to enhance plasmid DNA (pDNA) delivery. In this study, we complexed pDNA with combinations of FLR and FLH peptides, termed GET* nanoparticles. We sought to enhance our gene-activated scaffold platform by incorporating GET* nanoparticles into collagen–nanohydroxyapatite scaffolds with proven osteogenic capacity. GET* N/P 8 was shown to be the most effective formulation for delivery to MSCs in 2D. Furthermore, GET* N/P 8 nanoparticles incorporated into collagen–nanohydroxyapatite (coll–nHA) scaffolds at a 1:1 ratio of collagen:nanohydroxyapatite was shown to be the optimal gene-activated scaffold. pDNA encoding stromal-derived factor 1α (pSDF-1α), an angiogenic chemokine which plays a role in BMP mediated differentiation of MSCs, was then delivered to MSCs using our optimised gene-activated scaffold platform, with the aim of significantly increasing angiogenesis as an important precursor to bone repair. The GET* N/P 8 coll–nHA scaffolds successfully delivered pSDF-1α to MSCs, resulting in a significant, sustained increase in SDF-1α protein production and an enhanced angiogenic effect, a key precursor in the early stages of bone repair.

Graphene oxide-modified silk fibroin/nanohydroxyapatite scaffold loaded with urine-derived stem cells for immunomodulation and bone regeneration

BackgroundThe invasive and complicated procedures involving the use of traditional stem cells limit their application in bone tissue engineering. Cell-free, tissue-engineered bones often have complex scaffold structures and are usually engineered using several growth factors (GFs), thus leading to costly and difficult preparations. Urine-derived stem cells (USCs), a type of autologous stem cell isolated noninvasively and with minimum cost, are expected to solve the typical problems of using traditional stem cells to engineer bones. In this study, a graphene oxide (GO)-modified silk fibroin (SF)/nanohydroxyapatite (nHA) scaffold loaded with USCs was developed for immunomodulation and bone regeneration.MethodsThe SF/nHA scaffolds were prepared via lyophilization and cross-linked with GO using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxy succinimide (NHS). Scaffolds containing various concentrations of GO were characterized using scanning electron microscopy (SEM), the elastic modulus test, Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectrometer (XPS). Examinations of cell adhesion, proliferation, viability, morphology, alkaline phosphatase activity, and osteogenesis-related gene expression were performed to compare the osteogenesis-related biological behaviors of USCs cultured on the scaffolds. The effect of USC-laden scaffolds on the differentiation of macrophages was tested using ELISA, qRT-PCR, and immunofluorescence staining. Subcutaneous implantations in rats were performed to evaluate the inflammatory response of the USC-laden scaffolds after implantation. The scaffolds loaded with USCs were implanted into a cranial defect model in rats to repair bone defects. Micro-computed tomography (μCT) analyses and histological evaluation were performed to evaluate the bone repair effects.ResultsGO modification enhanced the mechanical properties of the scaffolds. Scaffolds containing less than 0.5% GO had good biocompatibility and promoted USC proliferation and osteogenesis. The scaffolds loaded with USCs induced the M2-type differentiation and inhibited the M1-type differentiation of macrophages. The USC-laden scaffolds containing 0.1% GO exhibited the best capacity for promoting the M2-type differentiation of macrophages and accelerating bone regeneration and almost bridged the site of the rat cranial defects at 12 weeks after surgery.ConclusionsThis composite system has the capacity for immunomodulation and the promotion of bone regeneration and shows promising potential for clinical applications of USC-based, tissue-engineered bones.

Comparative Study of Crystallization, Mechanical Properties, and In Vitro Cytotoxicity of Nanocomposites at Low Filler Loadings of Hydroxyapatite for Bone-Tissue Engineering Based on Poly(l-lactic acid)/Cyclo Olefin Copolymer.

A poly(l-lactic acid)/nanohydroxyapatite (PLLA/nHA) scaffold works as a bioactive, osteoconductive scaffold for bone-tissue engineering, but its low degradation rate limits embedded HA in PLLA to efficiently interact with body fluids. In this work, nano-hydroxyapatite (nHA) was added in lower filler loadings (1, 5, 10, and 20 wt%) in a poly(l-lactic acid)/cyclo olefin copolymer10 wt% (PLLA/COC10) blend to obtain novel poly(l-lactic acid)/cyclo olefin copolymer/nanohydroxyapatite (PLLA/COC10-nHA) scaffolds for bone-tissue regeneration and repair. Furthermore, the structure-activity relationship of PLLA/COC10-nHA (ternary system) nanocomposites in comparison with PLLA/nHA (binary system) nanocomposites was systematically studied. Nanocomposites were evaluated for structural (morphology, crystallization), thermomechanical properties, antibacterial potential, and cytocompatibility for bone-tissue engineering applications. Scanning electron microscope images revealed that PLLA/COC10-nHA had uniform morphology and dispersion of nanoparticles up to 10% of HA, and the overall nHA dispersion in matrix was better in PLLA/COC10-nHA as compared to PLLA/nHA. Fourier transformation infrared spectroscopy (FTIR), powder X-ray diffraction (XRD), and differential scanning calorimetry (DSC) studies confirmed miscibility and transformation of the α-crystal form of PLLA to the ά-crystal form by the addition of nHA in all nanocomposites. The degree of crystallinity (%) in the case of PLLA/COC10-nHA 10 wt% was 114% higher than pure PLLA/COC10 and 128% higher than pristine PLLA, indicating COC and nHA are acting as nucleating agents in the PLLA/COC10-nHA nanocomposites, causing an increase in the degree of crystallinity (%). Moreover, PLLA/COC10-nHA exhibited 140 to 240% (1–20 wt% HA) enhanced mechanical properties in terms of ductility as compared to PLLA/nHA. Antibacterial activity results showed that 10 wt% HA in PLLA/COC10-nHA showed substantial activity against P. aeruginosa, S. aureus, and L. monocytogenes. In vitro cytocompatibility of PLLA/COC10 and PLLA nanocomposites with nHA osteoprogenitor cells (MC3T3-E1) and bone mesenchymal stem cells (BMSC) was evaluated. Both cell lines showed two- to three-fold enhancement in cell viability and 10- to 30-fold in proliferation upon culture on PLLA/COC10-nHA as compared to PLLA/nHA composites. It was observed that the ternary system PLLA/COC10-nHA had good dispersion and interfacial interaction resulting in improved thermomechanical and enhanced osteoconductive properties as compared to PLLA/nHA.

A Naringin-loaded gelatin-microsphere/nano-hydroxyapatite/silk fibroin composite scaffold promoted healing of critical-size vertebral defects in ovariectomised rat

In this study, we investigated the effect of three-dimensional of naringin/gelatin microspheres/nano-hydroxyapatite/silk fibroin (NG/GMs/nHA/SF) scaffolds on repair of a critical-size bone defect of lumbar 6 in osteoporotic rats. In this work, a cell-free scaffold for bone-tissue engineering based on a silk fibroin (SF)/nano-hydroxyapatite (nHA) scaffold was developed. The scaffold was fabricated by lyophilization. Naringin (NG) was loaded into gelatin microspheres (GMs), which were encapsulated in the nHA/SF scaffolds. The materials were characterized using x ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy and thermogravimetric analysis. Moreover, the biomechanics, degradation, and drug-release profile of the scaffold were also evaluated. In vitro, the effect of the scaffold on the adhesion, proliferation, and osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) was evaluated. In vivo, at 3 months after ovariectomy, a critical-size lumbar defect was indued in the rats to evaluate scaffold therapeutic potential. A 3-mm defect in L6 developed in 60 SD rats, which were randomly divided into SF scaffold, nHA/SF scaffold, NG/nHA/SF scaffold, NG/GMs/nHA/SF scaffold, and blank groups (n = 12 each). At 4, 8, 12, and 16 weeks postoperatively, osteogenesis was evaluated by X-ray, micro-computed tomography, hematoxylin-eosin staining, and fast green staining, and by analysis of BMP-2, Runx2, and Ocn protein levels at 16 weeks. In our results, NG/GM/nHA/SF scaffolds exhibited good biocompatibility, biomechanical strength, and promoted BMSC adhesion, proliferation, and calcium nodule formation in vitro. Moreover, NG/GMs/nHA/SF scaffolds showed greater osteogenic differentiation potential than the other scaffolds in vitro. In vivo, gradual new bone formation was observed, and bone defects recovered by 16 weeks in the experimental group. In the blank group, limited bone formation was observed, and the bone defect was obvious. In conclusion, NG/GMs/nHA/SF scaffolds promoted repair of a lumbar 6 defect in osteoporotic rats. Therefore, the NG/GMs/nHA/SF biocomposite scaffold has potential as a bone-defect-filling biomaterial for bone regeneration.

Nano-Hydroxyapatite Scaffold Based on Recombinant Human Bone Morphogenetic Protein 2 and Its Application in Bone Defect Repair.

The best way in which to prepare scaffolds with good biological properties is an urgent problem in the field of tissue engineering. In this paper we discuss the preparation of nano-hydroxyapatite scaffold of recombinant human bone morphogenetic protein-2 (rhBMP-2) and its application in bone defect repair. rhBMP-2 reagent was dissolved in 1 mol/L sodium dihydrogen phosphate solution, and the rhBMP-2 solution was added to the nano-hydroxyapatite artificial bone with a 100 μL glass micro dropper at the rate of 10 drops/min to obtain Nano-HA/rhBMP-2 composite artificial bone. In in vivo experiments, rabbits were fixed on an operating table, a 2 cm longitudinal incision was made in the middle part of the radial forearm, and the radius was cut with a wire saw and periosteum, 2.5 cm away from the distal radius. After washing the wound with normal saline, Adv-hBMP-2/MC3T3-E1 nano-HA composite artificial bone, MC3T3-E1 nan-HA composite artificial bone, or Nano-HA artificial bone were implanted in different groups. The artificial bone scaffold prepared in this study has a stronger ability to repair bone defects than the alternatives, and is a promising prospect for the clinical treatment of bone defects.

Improved healing of critical-size femoral defect in osteoporosis rat models using 3D elastin/polycaprolactone/nHA scaffold in combination with mesenchymal stem cells

Osteoporosis is a common bone disease that results in elevated risk of fracture, and delayed bone healing and impaired bone regeneration are implicated by this disease. In this study, Elastin/Polycaprolactone/nHA nanofibrous scaffold in combination with mesenchymal stem cells were used to regenerate bone defects. Cytotoxicity, cytocompatibility and cellular morphology were evaluated in vitro and observations revealed that an appropriate environment for cellular attachment, growth, migration, and proliferation is provided by this scaffold. At 3 months following ovariectomy (OVX), the rats were used as animal models with an induced critical size defect in the femur to evaluate the therapeutic potential of osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs) seeded on 3 dimension (3D) scaffolds. In this experimental study, 24 female Wistar rats were equally divided into three groups: Control, scaffold (non-seeded BM-MSC), and scaffold + cell (seeded BM-MSC) groups. 30 days after surgery, the right femur was removed, and underwent a stereological analysis and RNA extraction in order to examine the expression of Bmp-2 and Vegf genes. The results showed a significant increase in stereological parameters and expression of Bmp-2 and Vegf in scaffold and scaffold + cell groups compared to the control rats. The present study suggests that the use of the 3D Elastin/Polycaprolactone (PCL)/Nano hydroxyapatite (nHA) scaffold in combination with MSCs may improve the fracture regeneration and accelerates bone healing at the osteotomy site in rats.

SCANINING ELECTRON MICROSCOPIC EVALUATION OF CHITOSAN NANOHYDROXYAPATITE SCAFFOLD FOR SOCKET HEALING IN RABBITS WITH INDUCED OSTEOPOROSIS

INTRODUCTION: Significant development has been achieved with bioceramics and biopolymer scaffolds in the construction of artificial bone. In the present study, Chitosan– nanohydroxyapatite (CS/nHA) scaffolds have been developed as bone graft substitutes in rabbits with induced osteoporosis.OBJECTIVES: To evaluate the effect of Chitosan- nanohydroxyapatite (CS/nHA) scaffold in bone tissue regeneration of extracted socket in rabbits with induced osteoporosis; using Scanning Electron Microscope and Energy Dispersive x-ray analysis [EDXA].MATERIALS AND METHODS: Twenty NewZealand white female rabbits were randomly divided into 2 equal groups; group A (osteoporosis) with no graft material, and group B (osteoporosis with CS/nHA). After the experimental period the animals were sacrificed at 2 & 4 weeks. The effect of CS/Nha was evaluated using Scanning Electron Microscope and Energy Dispersed x-ray analysis [EDXA]. The data obtained was tabulated and subjected to statistical analysis.RESULTS: Scanning Electron Microscopic results revealed more trabecular bone thickness with regular bone surface, and uniform osteocyte lacunae in the group B (osteoporosis with CS/nHA) than group A (osteoporosis), and these results were proved by EDXA which showed more Ca and P ratio in the group B than group A.CONCLUSIONS: CS/nHA scaffold is considered an effective hard tissue engineering material with sufficient interconnected porosity and mechanical strength to allow cell adhesion, migration, growth and proliferation resulting in good integration with surrounding tissues, and rapid bone healing, thus it might be used in osteoporotic condition.

Mesenchymal stem cell-loaded thermosensitive hydroxypropyl chitin hydrogel combined with a three-dimensional-printed poly(ε-caprolactone) /nano-hydroxyapatite scaffold to repair bone defects via osteogenesis, angiogenesis and immunomodulation.

Chitin-derived hydrogels are commonly used in bone regeneration because of their high cell compatibility; however, their poor mechanical properties and little knowledge of the interaction between the materials and host cells have limited their practical application.Methods: To evaluate osteoinductivity and enhance the mechanical properties of a newly synthesized thermosensitive hydroxypropyl chitin hydrogel (HPCH), a mesenchymal stem cell (MSC)-encapsulated HPCH was infused into a three-dimensional-printed poly (ε-caprolactone) (PCL)/ nano-hydroxyapatite (nHA) scaffold to form a hybrid scaffold. The mechanical properties and cell compatibility of the scaffold were tested. The interaction between macrophages and scaffold for angiogenesis and osteogenesis were explored in vitro and in vivo.Results: The hybrid scaffold showed improved mechanical properties and high cell viability. When MSCs were encapsulated in HPCH, osteo-differentiation was promoted properly via endochondral ossification. The co-culture experiments showed that the hybrid scaffold facilitated growth factor secretion from macrophages, thus promoting vascularization and osteoinduction. The Transwell culture proved that MSCs modulated the inflammatory response of HPCH. Additionally, subcutaneous implantation of MSC-encapsulated HPCH confirmed M2 activation. In situ evaluation of calvarial defects confirmed that the repair was optimal in the MSC-loaded HPCH + PCL/nHA group.Conclusions: PCL/nHA + HPCH hybrid scaffolds effectively promoted vascularization and osteoinduction via osteogenesis promotion and immunomodulation, which suggests promising applications for bone regeneration.