Bovine Bone Scaffold Research Articles

Advances in Regenerative and Reconstructive Medicine in the Prevention and Treatment of Bone Infections.

Reconstructive and regenerative medicine are critical disciplines dedicated to restoring tissues and organs affected by injury, disease, or congenital anomalies. These fields rely on biomaterials like synthetic polymers, metals, ceramics, and biological tissues to create substitutes that integrate seamlessly with the body. Personalized implants and prosthetics, designed using advanced imaging and computer-assisted techniques, ensure optimal functionality and fit. Regenerative medicine focuses on stimulating natural healing mechanisms through cellular therapies and biomaterial scaffolds, enhancing tissue regeneration. In bone repair, addressing defects requires advanced solutions such as bone grafts, essential in medical and dental practices worldwide. Bovine bone scaffolds offer advantages over autogenous grafts, reducing surgical risks and costs. Incorporating antimicrobial properties into bone substitutes, particularly with metals like zinc, copper, and silver, shows promise in preventing infections associated with graft procedures. Silver nanoparticles exhibit robust antimicrobial efficacy, while zinc nanoparticles aid in infection prevention and support bone healing; 3D printing technology facilitates the production of customized implants and scaffolds, revolutionizing treatment approaches across medical disciplines. In this review, we discuss the primary biomaterials and their association with antimicrobial agents.

Preosteoblast Adhesion and Viability Study of Freeze-Dried Bovine Bone Block Scaffold Coated with Human Umbilical Cord Mesenchymal Stem Cell Secretome.

Combining a three-dimensional scaffold with growth factors before implantation is one method used to increase scaffold bioactivity in bone tissue engineering. The mesenchymal stem cell (MSC)-conditioned medium (CM), called secretome, contains many proteins and growth factors required for tissue repair and growth. This study evaluated the bioactivity of a bovine bone scaffold combined with the secretome of human umbilical cord MSCs (hUC-MSCs) by analyzing MC3T3-E1 cell adhesion and viability on the scaffold. This in vitro laboratory study evaluated the effect of hUC-MSC secretome applied to bovine bone scaffolds processed using various techniques on MC3T3-E1 cell adhesion and viability. The three experimental groups included deproteinized bovine bone mineral-secretome (DBBM-CM), freeze-dried bovine bone-secretome (FDBB-CM), and decellularized FDBB-CM, whereas the control group was treated with DBBM alone. The cell adhesion test was performed using the centrifugation method after 6 and 24 hours, whereas the cell viability test was conducted using the trypan blue exclusion method after 24, 48, and 72 hours. Cell attachment was visualized after 4',6-diamidino-2-phenylindole staining and viewed under inverted fluorescence microscopy. Statistical analyses were performed using one-way analysis of variance, followed by a post hoc test in cases of significant differences. Statistical analyses showed significantly greater adhesion of the preosteoblasts to the FDBB-CM scaffold at 6 hours (p = 0.002). The results of the adhesion test at 24 hours and the viability tests at all observation times showed no significant differences (p > 0.05). This study found that the average MC3T3-E1 cell adhesions and viabilities were highest for the FDBB-CM and DBBM-CM scaffolds. DBBM scaffolds with the secretome had better cell adhesion and viability than those without the secretome. The addition of MSC secretome increased bovine bone scaffold bioactivity especially in DBBM and FDBB scaffolds.

Comparative Evaluation of the Differentiation and Proliferation Potential of Dental Pulp Stem Cells on Hydroxyapatite/Beta-Tricalcium Bone Graft and Bovine Bone Graft: An In Vitro Study.

Stem cells of mesenchymal origin have good proliferative capacity when compared to other stem cell types. Dental pulp stem cells (DPSCs) are a variety of mesenchymal cells obtained from the pulpal tissue of teeth and are abundantly available and easy to obtain. DPSCs facilitate and improve the formation of new bone using different bone graft scaffolds. This present study aims to evaluate and compare the osteogenic potential of DPSCs on alloplastic and xenogeneic bone grafts. Hydroxyapatite and beta-tricalcium bone graft and bovine bone graft were used in a triplicate manner in the laboratory. DPSCs were obtained from the pulpal tissue of extracted third molars in the laboratory. The cytotoxicity, osteogenic potential, and difference in the rate of proliferation of mesenchymal cells on the biomaterials were assessed. Darker purple staining was seen in the case of hydroxyapatite/beta-tricalcium bone graft on MTT colorimetric assay stating that there was an increase in cell viability in hydroxyapatite/beta-tricalcium bone graft as compared to the bovine bone graft. Hydroxyapatite/beta-tricalcium bone graft showed more osteogenic potential as compared to the bovine bone graft as a higher degree of red staining was seen in Alizarin staining. Higher cell viability and higher osteogenic proliferation and differentiation were seen on the hydroxyapatite/beta-tricalcium bone graft compared to the bovine bone scaffold.

Correction: Gonçalves et al. Antimicrobial Activity of Bovine Bone Scaffolds Impregnated with Silver Nanoparticles on New Delhi Metallo-β-Lactamase-Producing Gram-Negative Bacilli Biofilms. Compounds 2023, 3, 584–595

The author would like to make the following corrections to the original publication [...]

The Influence of Different Pore Sizes of Bovine Bone Scaffold Material on the Oral Microbiota

Bone scaffolds are a kind of bone repair material, but bone scaffold infection is a common and serious clinical problem that has not been well studied in the context of the oral microbiota. We prepared two different bone scaffold materials with different pore sizes from adult and fetal bovine cancellous bones and compared them with hydroxyapatite (HA) powder. After incubation, we facilitated the initial colonization by the oral microbiota on the three materials by using the saliva of healthy people. Through high-throughput sequencing and analysis, we found that the different pore sizes of the bone scaffold materials had a certain influence on the initial colonization by the oral microbiota, affecting β-diversity and functional profiles. Redundancy analysis (RDA) and cooccurrence analysis indicated that initial microbial colonization by the oral microbiota in fetal bone scaffold materials with smaller pore sizes was more likely to cause infection. In addition, there may be some functional bacteria that play an interactive role in early colonization by the oral microflora in bone scaffold materials, such as those of the genera Bacillus and Lactobacillus.

Osteoinductive and Osteogenic Capacity of Freeze-Dried Bovine Bone Compared to Deproteinized Bovine Bone Mineral Scaffold in Human Umbilical Cord Mesenchymal Stem Cell Culture: An In Vitro Study.

Freeze-dried bovine bone scaffold (FDBB) or decellularized FDBB (dc-FDBB) was developed as an ideal scaffold with osteoinductive properties. This research aims to compare the osteoinductive properties marked by the expression of runt-related transcription factor-2 (RUNX2) and Osterix (OSX) and the osteogenic capacity of these scaffolds imbued with human umbilical cord mesenchymal stem cells (hUCMSCs). This study was performed in five experimental groups: a negative control group (C-) of hUCMSCs with a normal growth medium, a positive control group (C + ) of hUCMSCs with an osteogenic medium, experimental group 1 (E1) with an FDBB conditioned medium (CM), and experimental group 2 (E2) with a dc-FDBB-CM, and a third experimental group (E3) consisting of a DBBM-CM. Alizarin red staining was performed to qualitatively assess osteoinductive capacity. RUNX2 and OSX expression was quantified using real-time quantification polymerase chain reaction with two replications on day six (D6) and day 12 (D12) as fold changes. This experiment revealed that hUCMSCs were positively expressed by CD73, CD90, and CD105 but were not expressed by CD34. Alizarin red staining showed that E1 had the most calcium deposition on D6 and D12, followed by E3 and then E2 The RUNX2 and OSX expression was higher in E1 but this difference was not significant. The OSX expression in E1,E2,E3 was lower on D12 and C+ of OSX had the highest expression. There was a significant difference of fold change measured between all groups (p < 0.05), and there was no significant difference between any of the groups treated with OSX and RUNX2 on D6 and D12. FDBB osteoinduction and osteogenic capacity were higher when compared with DBBM and dc-FDBB.

The Effect of Low-Temperature Thermal Processing on Bovine Hydroxyapatite Bone Substitutes, toward Bone Cell Interaction and Differentiation.

Ideal bone grafting scaffolds are osteoinductive, osteoconductive, and encourage osteogenesis through the remodeling processes of bone resorption, new bone formation, and successful integration or replacement; however, achieving this trifecta remains challenging. Production methods of bone grafts, such as thermal processing, can have significant effects on the degree of cell-surface interactions via wide-scale changes in the material properties. Here, we investigated the effects of small incremental changes at low thermal processing temperatures on the degree of osteoclast and osteoblast attachment, proliferation, and differentiation. Bovine bone scaffolds were prepared at 100, 130, 160, 190, and 220 °C and compared with a commercial control, Bio-Oss®. Osteoclast attachment and activity were significantly higher on lower temperature processed bone and were not present ≥190 °C. The highest osteoblast proliferation and differentiation were obtained from treatments at 130 and 160 °C. Similarly, qRT2-PCR assays highlighted osteoblasts attached to bone processed at 130 and 160 °C as demonstrating the highest osteogenic gene expression. This study demonstrated the significant effects of small-scale processing changes on bone graft materials in vitro, which may translate to a tailored approach of cellular response in vivo.

Collagen matrices are preserved following decellularization of a bovine bone scaffold.

The decellularization of bovine bone has emerged as a strategy for the repair, replacement, and regeneration of bone defects. To evaluate the effects of a new protocol of bone decellularization and its impact on the structure and collagen scaffold. Cancellous bone from bovine femur was dissected in fragments and decellularized based on protocol of multiple steps. The residual protein levels, histological, morphometric, and scanning electronic microscopy analyses were carried out to evaluate the effects of decellularization and the impact on the structure and collagen scaffold. A cytotoxicity assay was performed. Residual protein analysis showed an important removal of bone marrow components and cell debris from the bone. Sections revealed that collagen fibers presented integrity and absence of cells in the decellularized bone. Sirius Red-stained sections of collagen fiber collagen matrix were maintained after decellularization. Scanning electron microscopy revealed that the main bone structure, despite being irregular, was maintained in both groups, with no significant visual differences between the surface characteristics according to the groups. Decellularized bovine bone demonstrated a degree of toxicity of 3, indicating moderate reactivity. The present data demonstrate that the main bone structure was maintained. Additionally, the chemical and physical treatments were able to remove cellular debris, and extracellular matrix architecture and collagen were preserved. However, the tissue showed moderate toxicity.

Apatite-forming ability of bioglass coated bovine bone scaffolds

The aim of this study was to investigate the bone-like apatite forming ability of bioglass coated bones by immersing them in simulated body fluid (SBF). Various types of bioactive coatings with an average thickness of 15 ?m were deposited on the surface of the cortical bovine bone by vacuum assisted infiltration.More intense apatite crystallization was observed using additives such as hydroxyapatite, sintered bone and ?-whitlockite in the base glass. The dissolution properties of the scaffolds were determined by ICP-OES and XRF methods. The consequence of the release of Ca and P during immersion in SBF on the mechanical properties was observed through measurements of microhardness, bending and compressive strength.

Comparing the Effects of Alpha-Tocopherol and Tocotrienol Isomers on Osteoblasts hFOB 1.19 Cultured on Bovine Bone Scaffold

Tocotrienol mixtures have been shown to exert anabolic actions on the skeletal system in animal studies, but it is unclear which tocotrienol isomer shows the most prominent effects. This study aims to investigate the most active tocotrienol isomers using hFOB 1.19 human osteoblasts cultured on a bovine bone scaffold. The bovine trabecular bone was sectioned, demineralised and freeze-dried to form the scaffold. hFOB 1.19 osteoblasts were cultured on the bone scaffolds in humidified condition at 37 °C and 5% carbon dioxide with vitamin E isomers (alpha-, beta-, gamma-, delta-tocotrienol and alpha-tocopherol). The cell differentiation capacity of tocotrienol isomers was investigated through morphological observation, alkaline phosphatase (ALP) activity and osteocalcin expression. Changes in the bone scaffolds were determined using histomorphometry methods. Osteoblast culture treated with gamma- and delta-tocotrienols showed a significant increase in ALP activity and osteocalcin expression. Bone structural histomorphometry analysis showed that bone scaffolds treated with gamma- and delta-tocotrienol showed significant increases in bone volume and trabecular thickness. In conclusion, gamma- and delta-tocotrienol show the most prominent bone anabolic effects by increasing osteoblast differentiation and enhancing bone microstructure.

Skeletal microenvironment system utilising bovine bone scaffold co-cultured with human osteoblasts and osteoclast-like cells.

A three-dimensional ex vivo bone cell culture system mimicking the skeletal system is useful for bone tissue engineering and as drug discovery platforms. The present study aimed to establish a three-dimensional skeletal culture system using native bovine bone scaffolds and human bone cells. Bovine bone scaffolds were cultured with human foetal osteoblasts 1.19 and human peripheral blood mononuclear cells for 21 days under standard culture conditions. The following groups were established: Decalcified unseeded bone scaffold (DUBS) as baseline control, decalcified seeded bone scaffold (DSBS) to mimic osteoporosis condition and undecalcified seeded bone scaffold to mimic normal condition. The scaffold's porosity and cell attachment on the scaffolds were determined using scanning electron microscopy. Histological evaluation was used to examine changes in trabecular bone structure. Dual-energy X-ray absorptiometry analysis was performed to determine the bone mineral density (BMD) and bone mineral content (BMC) of the scaffolds. A compression test was performed to examine the total biomechanical strength of the scaffolds. The trabecular thickness and number increased, while the trabecular separationwas reduced slightly in DSBS than in DUBS (P>0.05). The BMD and BMC increased significantly (P<0.05), while the compressive strength only increased slightly in DSBS than in DUBS (P>0.05). In conclusion, the ex vivo skeletal microenvironment comprising native bovine bone scaffolds seeded with bone cells is structurally, functionally and mechanically comparable with natural bone. This system may be used as a platform to understand bone physiology and screen for potential drug candidates.

Design of two natural deproteinized bovine bone scaffolds and evaluation of the effect of initial cell seeding density on repairing bone defects.

Engineering functional bone using combinations of cells, scaffolds, and bioactive factors is a promising strategy for the bone-tissue regeneration, while challenge remains. Chemical methods deproteinizing natural bovine cancellous bone to remove immunogenic are poorly understood, and the cell seeding density to promote bone formation still needs to be clarified. In this study, 8.0 × 8.0 × 2.0 mm bovine cancellous bones were either treated with H2 O2 for 8hr or pepsin for 24 hr and then inoculated with MC3T3-E1 osteoblasts with two cell densities (1 × 106 cells/ml or 4 × 106 cells/ml)separately. We compared the appearance of the bones treated by the two chemical deproteinizing methods, as well as the proliferation ability of the inoculating cell density at 1 × 106 cells/ml. Moreover, scanning electron microscopy was done to analyze the growth of cells on the surface of the material, and an alkaline phosphatase assay was performed to assess osteogenic differentiation. We showed that both treated bones treatments are biocompatible, but bones treated with H2 O2 were more conducive to osteoblast differentiation and ALP secretion, especially when seeded at the higher cell density at 4 × 106 cells/ml. We concluded that chemical deproteinized bovine cancellous bones met the basic bone graft material requirements. Cell seeding density is an important factor to promote the material's osteogenic ability, with H2 O2 -deproteinized bones exhibiting enhanced osteoblast differentiation.

The construction of a novel xenograft bovine bone scaffold, (DSS)6-liposome/CKIP-1 siRNA/calcine bone and its osteogenesis evaluation on skull defect in rats.

BackgroundXenograft bone scaffolds have advantages such as mechanical strength, sufficient source and safety. Combined with siRNA properly targeting CKIP-1, a negative regulator of osteogenesis, may contribute to the repair result of calcine bone alone.MethodsHerein, we constructed a novel xenograft bovine bone scaffold namely (DSS)6-liposome/CKIP-1 siRNA/calcine bone, the characteristics of which were investigated by confirming the effect of (DSS)6-liposome, observing the appearance and testing mechanical strength of calcine bone, and observing the combined result of CKIP-1 siRNA by FAM immunofluorescence. In addition, cytotoxicity by CCK-8 and LDH activity of L929 ​cells and MC3T3-E1 osteoblasts cultured with the scaffold were tested in vitro, primary osteoblasts proliferation, the mRNA expressions of CKIP-1, ALP, COL1-α and OCN, the protein expressions of CKIP-1, BMP-2, COL-1 and Runx2 and calcium nodules were also determined by CCK-8, RT-qPCR, western-blot and Alizarin Red staining in vitro. Then, we successively established the skull defect model for evaluating the repair result of the novel scaffold by HE staining of 2, 4, 8 and 12 weeks, immumohistochemical stainings of 2, 4, 8 and 12 weeks such as ALP, COL-1α and OCN, Mirco-CT scanning of 4 and 12 weeks and the relative parameters and so on in vivo.ResultsIt indicated that (DSS)6-liposome/CKIP-1 siRNA/calcine bone could successfully knock down the CKIP-1 mRNA and protein expressions, promote osteoblasts proliferation with the little cytotoxicity in vitro, increase the protein expressions of BMP-2, COL-1 and Runx2 in vitro, increase mRNA expressions of ALP, COL-1α and OCN in vitro and in vivo, and have a better bone defect repair effect with few side effects in rats after 12 weeks.ConclusionOur research indicates (DSS)6-liposome/CKIP-1 siRNA/calcine bone could repair skull defects well in rats, and it may lay the foundation of applicating the novel xenograft bone scaffold in the clinical.The Translational potential of this articleThese findings provide evidence that (DSS)6- liposome/CKIP-1 siRNA/calcine bone could be used as a novel xenograft bone scaffold for osteogenesis with the good safety.

Lithium-incorporated deproteinized bovine bone substitute improves osteogenesis in critical-sized bone defect repair.

This study aimed to explore the surface modification of deproteinized bovine bone using lithium-ion and evaluate its efficacy on osteogenesis improvement and critical-sized bone defect repair. Hydrothermal treatment was performed to produce lithium-incorporated deproteinized bovine bone. In vitro study, human osteosarcoma cell MG63 (MG63) was cultured with the bone substitute to evaluate the cell viability and then calcium deposition was measured to analyze the osteogenesis. In vivo studies, male adult goats were chosen to build critical-sized bone defect model and randomly divided into three groups. The goats were treated with autogenous cancellous bone, lithium-incorporated deproteinized bovine bone, and deproteinized bovine bone. Animals were evaluated using radiological analysis including X-ray, computed tomography, and Micro-CT; histological methods involving hematoxylin-eosin dyeing, Masson dyeing, and immunofluorescence detection at 4 and 12 weeks after surgery were carried out. According to the results, lithium-incorporated deproteinized bovine bone produced nano-structured surface layer. The lithium-incorporated deproteinized bovine bone could promote the osteoblast proliferation and increase the calcium deposition. In vivo studies, radiographic results revealed that lithium-incorporated deproteinized bovine bone scaffolds provided better performance in terms of mean gray values of X films, mean pixel values of computed tomography films, and bone volume and trabecular thickness of micro-computed tomography pictures when compared with the deproteinized bovine bone group. In addition, histological analysis showed that the lithium-incorporated deproteinized bovine bone group also significantly achieved larger new bone formation area. At the same time, when the expression of osteogenic factors in vivo was evaluated, runt-related transcription factor 2 (Runx2) and collagen type one (Col-1) were expressed more in lithium-incorporated deproteinized bovine bone group than those in deproteinized bovine bone group. However, the bone defect repair effect using autograft is still a little better than that of lithium-incorporated deproteinized bovine bone substitute based on our results. In conclusion, surface lithium-incorporated deproteinized bovine bone achieved improvement of osteogenesis effect and could enhance the new bone formation in critical-sized bone defects.

Biomimetic matrix fabricated by LMP-1 gene-transduced MC3T3-E1 cells for bone regeneration

Bone healing is regulated by multiple microenvironmental signals provided by the extracellular matrix (ECM). This study aimed to mimic the native osteoinductive microenvironment by developing an ECM using gene-transduced cells. The LIM mineralization protein-1 (LMP-1) gene was transferred to murine pre-osteoblast cells (MC3T3-E1) using lentiviral vectors. Western blotting assay indicated that the MC3T3-E1 cells expressed an increased level of bone morphologic protein-2, -4 and -7 (BMP-2, -4 and -7) after LMP-1 gene transduction. The transduced cells were then seeded into calcined bovine bone scaffolds and cultured for 7, 14, and 21 days to construct ECMs on the scaffolds. The ECM–scaffold composites were then decellularized using the freeze-drying method. Scaffolds without ECM deposition were used as controls. The composites and controls were implanted into critical-sized bone defects created in the distal femurs of New Zealand rabbits. Twelve weeks after the surgery, both microcomputed tomography and histologic results indicated that the 7-day-cell-modified ECM–scaffold composites induced bone regeneration with significantly larger volume, trabecular thickness and connectivity than the controls. However, the 14- and 21-day-cell-modified ECM–scaffold composites triggered sustained inflammation response even at 12 weeks after the surgery and showed less bone ingrowth and integration than their 7-day-cell-modified counterparts. In conclusion, these results highlight the viable gene transfer techniques for manipulating cells in a constructed microenvironment of ECM for bone regeneration. However, the unresolved inflammation relating to the duration of ECM modification needs to be considered.

Stanozolol-soaked grafts enhance new bone formation in rat calvarial critical-size defects

Androgen hormones play a significant role in regulating bone morphogenesis and in maintaining bone homeostasis throughout life. This study aimed to investigate the local effects of the non-aromatizable androgen stanozolol (ST) on bone regeneration in rats. Bilateral critical-size defects were created in the parietal bone of 26 male Wistar rats: the defect on one side was filled with a deproteinized bovine bone scaffold (DBB) soaked in ST solution (test) and the contralateral with DBB alone (control). Samples were collected at one month and three months. Histomorphometry revealed a significantly higher new bone formation (NB) (24.41% ± 4.14% versus 15.01% ± 2.43%, p < 0.05) and mineral apposition rate (MAR) (9.20 μm/day ± 0.37 versus 6.50 μm/day ± 1.09, p < 0.05) in the test versus control group at one month. Accordingly, real time-polymerase chain reaction revealed a consistently higher Runx2 expression in test samples (fold change test/control: 4.50 ± 1.17, p ≤ 0.05). No morphometrical differences between groups were detected at three months (p > 0.05). However, test samples were characterized by an increase in blood capillary density from one month (11.43 n mm−2 ± 2.01) to three months (28.26 n mm−2 ± 5.62), providing evidence of a vital remodeling tissue. Control samples presented a decrease of anti-Osterix (SP7)/anti-osteocalcin (BGLAP) (3.9 n mm−2 ± 0.32 versus 1.01 n mm−2 ± 0.20) and alkaline phosphatase (ALP) (12.14 n mm−2 ± 6.29 versus 6.29 n mm−2 ± 2.73) immunohistochemical-positive elements, which was suggestive of a stabilized healing phase. Based on these observations, local ST administration boosted bone regeneration in rat calvarial critical-size defects at one month. This study showed the potential of local steroid delivery in bone regeneration.

Ultrastructural evaluation of mesenchymal stem cells from inflamed periodontium in different in vitro conditions.

This research aimed to observe the behavior of mesenchymal stem cells (MSCs) isolated from periodontal granulation tissue (gt) when manipulated ex vivo to induce three-dimensional (3D) spheroid (aggregates) formation as well as when seeded on two bone scaffolds of animal origin. Periodontal gt was chosen as a MSC source because of its availability, considering that it is eliminated as a waste material during conventional surgical therapies. 3D aggregates of cells were generated; they were grown for 3 and 7 days, respectively, and then prepared for transmission electron microscopic analysis. The two biomaterials were seeded for 72 h with gtMSCs and prepared for scanning electronic microscopic observation. The ultrastructural analysis of 3D spheroids remarked some differences between the inner and the outer cell layers, with a certain commitment observed at the inner cells. Both scaffolds showed a relatively smooth surface at low magnification. Macro- and micropores having a scarce distribution were observed on both bone substitutes. gtMSCs grew with relative difficulty on the biomaterials. After 72 h of proliferation, gtMSCs scarcely covered the surface of bovine bone scaffolds, demonstrating fibroblast-like or star-like shapes with elongated filiform extensions. Our results add other data on the possible usefulness of gtMSC and could question the current paradigm regarding the complete removal of chronically inflamed gts from the defects during periodontal surgeries. Until optimal protocols for ex vivo manipulation of MSCs are available for clinical settings, it is advisable to use biocompatible bone substitutes that allow the development of progenitor cells.

In Vitro Study of Surface Modified Poly(ethylene glycol)-Impregnated Sintered Bovine Bone Scaffolds on Human Fibroblast Cells.

Scaffold design from xenogeneic bone has the potential for tissue engineering (TE). However, major difficulties impede this potential, such as the wide range of properties in natural bone. In this study, sintered cortical bones from different parts of a bovine-femur impregnated with biodegradable poly(ethylene glycol) (PEG) binder by liquid phase adsorption were investigated. Flexural mechanical properties of the PEG-treated scaffolds showed that the scaffold is stiffer and stronger at a sintering condition of 1000°C compared with 900°C. In vitro cytotoxicity of the scaffolds evaluated by Alamar Blue assay and microscopic tests on human fibroblast cells is better at 1000°C compared with that at 900°C. Furthermore, in vitro biocompatibility and flexural property of scaffolds derived from different parts of a femur depend on morphology and heat-treatment condition. Therefore, the fabricated scaffolds from the distal and proximal parts at 1000°C are potential candidates for hard and soft TE applications, respectively.

Porous deproteinized bovine bone scaffold with three-dimensional localized drug delivery system using chitosan microspheres.

BackgroundBone substation grafts, such as hydroxyapatite (HA) and tricalciumphosphate (TCP), have been extensively used in clinical applications, but evidence suggests that they offer poor osteoinductive properties compared to allografts and autografts. In order to increase bone growth with such grafts, Bone Morphogenetic Protein 2 (BMP-2) was incorporated into a three dimensional reservoir. The purpose of the present study was to develop a novel drug delivery system which is capable of controlled release of BMP-2.MethodsDBB were prepared from bovine cancellous bone harvested from fetal bovine femur or tibia and then sinting at 1000°C. BMP-2-loaded chitosan (CS) microspheres were fabricated by cross-linking. Then the treated DBB powders were blended with chitosan microspheres solution. Finally, the composites were lyophilized with a freeze dryer to obtain the DBB/CMs scaffolds. X-ray diffractor (XRD), scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR) were used to characterize the sample. The quantification of the delivery profile of BMP-2 was determined using an enzyme-linked immunosorbent assay (ELISA) kit. The in vitro assays were to characterize the biocompatibility of this composite.ResultsIn this study, BMP-2/Chitosan (CS) microspheres were successively loaded onto a deproteinized bovine bone (DBB) scaffold. The release profile of BMP-2 indicated an initial burst release followed by a more even sustained release. An in vitro bioactivity assay revealed that the encapsulated growth factor was biologically active.ConclusionsThe cell culture assay suggest that the excellent biocompatibility of the DBB- BMP-2/CS. Therefore, this novel microsphere scaffold system can be effectively used in current tissue engineering applications.

A Novel Deproteinized Bovine Bone Scaffold Containing Chitosan Microspheres for Controlled Delivery of BMP-2

The bone substation grafts, such as hydroxyapatite, TCP, have been used in the clinical. But it is found that they are not capable of enhancing bone regeneration due to their poor osteoinductive compared with allograft and autograft. In order to increase the osteoinductive, bone morphogenetic protein 2 (BMP-2) has highlighted to incorporate into a three dimensional reservoir. The purpose of the present study was to develop a novel drug delivery system with the capability of controlled releasing BMP-2. In this study, BMP-2/Chitosan (CS) microspheres were succeeded loaded on the deproteinized bovine bone (DBB) scaffold. The release profile of BMP-2 study indicated an initial burst release followed by a sustained release. In vitro bioactivity assay revealed that the encapsulated growth factor was biologically active as evidenced by cell culture. Therefore, the novel strategy of this microspheres-scaffold system is potentially to utilize the bone tissue engineered.