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1.
J Mater Sci Mater Med ; 27(9): 138, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27530301

RESUMO

In the past, bioactive bone cement was investigated in order to improve the durability of cemented arthroplasties by strengthening the bone-cement interface. As direct bone-cement bonding may theoretically lead to higher stresses within the cement, the question arises, whether polymethylmethacrylate features suitable mechanical properties to withstand altered stress conditions? To answer this question, in vivo experiments and finite element simulations were conducted. Twelve rabbits were divided into two groups examining either bioactive polymethylmethacrylate-based cement with unchanged mechanical properties or commercially available polymethylmethacrylate cement. The cements were tested under load-bearing conditions over a period of 7 months, using a spacer prosthesis cemented into the femur. For the finite element analyses, boundary conditions of the rabbit femur were simulated and analyses were performed with respect to different loading scenarios. Calculations of equivalent stress distributions within the cements were applied, with a completely bonded cement surface for the bioactive cement and with a continuously interfering fibrous tissue layer for the reference cement. The bioactive cement revealed good in vivo bioactivity. In the bioactive cement group two failures (33 %), with complete break-out of the prosthesis occurred, while none in the reference group. Finite element analyses of simulated bioactive cement fixation showed an increase in maximal equivalent stress by 49.2 to 109.4 % compared to the simulation of reference cement. The two failures as well as an increase in calculated equivalent stress highlight the importance of fatigue properties of polymethylmethacrylate in general and especially when developing bioactive cements designated for load-bearing conditions.


Assuntos
Cimentos Ósseos/química , Prótese de Quadril , Polimetil Metacrilato/química , Animais , Materiais Biocompatíveis , Fêmur/cirurgia , Análise de Elementos Finitos , Vidro , Teste de Materiais , Ortopedia , Coelhos , Estresse Mecânico , Suporte de Carga
2.
Med Sci Monit ; 20: 1942-9, 2014 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-25317537

RESUMO

BACKGROUND: The aim of the current study was to measure and compare the effect of various biomaterials for the healing of osteoporotic bone defects in the rat femur using 18F-sodium fluoride dPET-CT. MATERIAL AND METHODS: Osteoporosis was induced by ovariectomy and a calcium-restricted diet. After 3 months, rats were operated on to create a 4-mm wedge-shaped defect in the distal metaphyseal femur. Bone substitution materials of calcium phosphate cement (CPC), composites of collagen and silica, and iron foams with interconnecting pores were inserted. Strontium or bisphosphonate, which are well known for having positive effects in osteoporosis treatment, were added into the materials. Eighteen weeks after osteoporosis induction and 6 weeks following femoral surgery, dPET-CT studies scan were performed with 18F-Sodium Fluoride. Standardized uptake values (SUVs) and a 2-tissue compartmental learning-machine model (K1-k4, vessel density [VB], influx [ki]) were used for quantitative analysis. RESULTS: k3, reflecting the formation of fluoroapatite, revealed a statistically significant increase at the biomaterial-bone interface due to the Sr release from strontium-modified calcium phosphate cement (SrCPC) compared to CPC, which demonstrated enhanced new bone formation. In addition, k3 as measured in the porous scaffold silica/collagen xerogel (Sc-B30), showed a significant increase based on Wilcoxon rank-sum test (p<0.05) as compared with monolithic silica/collagen xerogel (B30) in the defect region. Furthermore, ki, reflecting the net plasma clearance of tracer to bone mineral measured in the iron foam with coating of the bisphosphonate zoledronic acid (Fe-BP), was enhanced as compared with plain iron foam (Fe) in the defect region. CONCLUSIONS: k3 was the most significant parameter for the characterization of healing processes and revealed the best differentiation between the 2 different biomaterials. PET scanning using 18F-sodium fluoride seems to be a sensitive and useful method for evaluation of bone healing after replacement with these biomaterials.


Assuntos
Materiais Biocompatíveis , Osteoporose/patologia , Fluoreto de Sódio/administração & dosagem , Animais , Modelos Animais de Doenças , Feminino , Imagem Multimodal , Tomografia por Emissão de Pósitrons , Ratos , Ratos Sprague-Dawley , Tomografia Computadorizada por Raios X
3.
Nanoscale Adv ; 6(3): 1011-1022, 2024 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-38298585

RESUMO

Non-cement pastes in the form of injectable materials have gained considerable attention in non-invasive regenerative medicine. Different osteoconductive bioceramics have been used as the solid phase of these bone pastes. Mesoporous bioactive glass can be used as an alternative bioceramic for paste preparation because of its osteogenic qualities. Plant-derived osteogenic agents can also be used in paste formulation to improve osteogenesis; however, their side effects on physical and physicochemical properties should be investigated. In this study, nano-bioactive glass powder was synthesized by a sol-gel method, loaded with different amounts of quercetin (0, 100, 150, and 200 µM), an antioxidant flavonoid with osteogenesis capacity. The loaded powder was then homogenized with a mixture of hyaluronic acid and sodium alginate solution to form a paste. We subsequently evaluated the rheological behavior, injectability, washout resistance, and in vitro bioactivity of the quercetin-loaded pastes. The washout resistance was found to be more than 96% after 14 days of immersion in simulated body fluid (SBF) as well as tris-buffered and citric acid-buffered solutions at 25 °C and 37 °C. All pastes exhibited viscoelastic behavior, in which the elastic modulus exceeded the viscous modulus. The pastes displayed shear-thinning behavior, in which viscosity was more influenced by angular frequency when the quercetin content increased. Results indicated that injectability was much improved using quercetin and the injection force was in the range 20-150 N. Following 14 days of SBF soaking, the formation of a nano-structured apatite phase on the surfaces of quercetin-loaded pastes was confirmed through scanning electron microscopy, X-ray diffractometry, and Fourier-transform infrared spectroscopy. Overall, quercetin, an antioxidant flavonoid osteogenic agent, can be loaded onto the nano-bioactive glass/hyaluronic acid/sodium alginate paste system to enhance injectability, rheological properties, and bioactivity.

4.
Polymers (Basel) ; 16(9)2024 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-38732723

RESUMO

A promising therapeutic option for the treatment of critical-size mandibular defects is the implantation of biodegradable, porous structures that are produced patient-specifically by using additive manufacturing techniques. In this work, degradable poly(DL-lactide) polymer (PDLLA) was blended with different mineral phases with the aim of buffering its acidic degradation products, which can cause inflammation and stimulate bone regeneration. Microparticles of CaCO3, SrCO3, tricalcium phosphates (α-TCP, ß-TCP), or strontium-modified hydroxyapatite (SrHAp) were mixed with the polymer powder following processing the blends into scaffolds with the Arburg Plastic Freeforming 3D-printing method. An in vitro degradation study over 24 weeks revealed a buffer effect for all mineral phases, with the buffering capacity of CaCO3 and SrCO3 being the highest. Analysis of conductivity, swelling, microstructure, viscosity, and glass transition temperature evidenced that the mineral phases influence the degradation behavior of the scaffolds. Cytocompatibility of all polymer blends was proven in cell experiments with SaOS-2 cells. Patient-specific implants consisting of PDLLA + CaCO3, which were tested in a pilot in vivo study in a segmental mandibular defect in minipigs, exhibited strong swelling. Based on these results, an in vitro swelling prediction model was developed that simulates the conditions of anisotropic swelling after implantation.

5.
J Pers Med ; 13(3)2023 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-36983646

RESUMO

For sinus grafting, different methods and materials are available. One possible shortcoming of particulate bone grafts is either overfilling or augmenting the planned implant area insufficiently. To overcome this risk and to determine the implant position prior augmentation, we present an approach using three-dimensional printed scaffolds. A patient with a remaining anterior dentition and bilateral severely atrophied posterior maxilla was seeking oral rehabilitation. The cone beam computed tomography (CBCT) showed residual bone heights between one and two millimeters. Following the three-dimensional reconstruction of the CBCT data, the positions of the implants were determined in areas 16 and 26. Three-dimensional scaffolds adapted to the topography of the sinus were virtually designed and printed using a calcium phosphate cement paste. Bilateral sinus floor augmentation applying the printed scaffolds with an interconnecting porosity followed. After nine months, a satisfying integration of the scaffolds was obvious. At the re-entry, vital bone with sufficient blood supply was found. One implant could be placed in positions 16 and 26, respectively. After five months, the implants could be uncovered and were provided with a temporary denture. The application of three-dimensionally printed scaffolds from calcium phosphate cement paste seems to be a promising technique to graft the severely atrophied posterior maxilla for the placement of dental implants.

6.
Biomater Sci ; 11(16): 5590-5604, 2023 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-37403758

RESUMO

Their excellent mechanical properties, degradability and suitability for processing by 3D printing technologies make the thermoplastic polylactic acid and its derivatives favourable candidates for biomaterial-based bone regeneration therapies. In this study, we investigated whether bioactive mineral fillers, which are known to promote bone healing based on their dissolution products, can be integrated into a poly(L-lactic-co-glycolic) acid (PLLA-PGA) matrix and how key characteristics of degradation and cytocompatibility are influenced. The polymer powder was mixed with particles of CaCO3, SrCO3, strontium-modified hydroxyapatite (SrHAp) or tricalcium phosphates (α-TCP, ß-TCP) in a mass ratio of 90 : 10; the resulting composite materials have been successfully processed into scaffolds by the additive manufacturing method Arburg Plastic Freeforming (APF). Degradation of the composite scaffolds was investigated in terms of dimensional change, bioactivity, ion (calcium, phosphate, strontium) release/uptake and pH development during long-term (70 days) incubation. The mineral fillers influenced the degradation behavior of the scaffolds to varying degrees, with the calcium phosphate phases showing a clear buffer effect and an acceptable dimensional increase. The amount of 10 wt% SrCO3 or SrHAp particles did not appear to be appropriate to release a sufficient amount of strontium ions to exert a biological effect in vitro. Cell culture experiments with the human osteosarcoma cell line SAOS-2 and human dental pulp stem cells (hDPSC) indicated the high cytocompatibility of the composites: For all material groups cell spreading and complete colonization of the scaffolds over the culture period of 14 days as well as an increase of the specific alkaline phosphatase activity, typical for osteogenic differentiation, were observed.


Assuntos
Osteogênese , Alicerces Teciduais , Humanos , Alicerces Teciduais/química , Glicóis , Fosfatos de Cálcio/química , Minerais , Diferenciação Celular , Estrôncio/química , Impressão Tridimensional
7.
Biomacromolecules ; 13(4): 1059-66, 2012 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-22364350

RESUMO

Biomimetic mineralization of collagen is an advantageous method to obtain resorbable collagen/hydroxy-apatite composites for application in bone regeneration. In this report, established procedures for mineralization of bovine collagen were adapted to a new promising source of collagen from salmon skin challenged by the low denaturation temperature. Therefore, in the first instance, variation of temperature, collagen concentration, and ionic strength was performed to reveal optimized parameters for fibrillation and simultaneous mineralization of salmon collagen. Porous scaffolds from mineralized salmon collagen were prepared by controlled freeze-drying and chemical cross-linking. FT-IR analysis demonstrated the mineral phase formed during the preparation process to be hydroxyapatite. The scaffolds exhibited interconnecting porosity, were sufficiently stable under cyclic compression, and showed elastic mechanical properties. Human mesenchymal stem cells were able to adhere to the scaffolds, cell number increased during cultivation, and osteogenic differentiation was demonstrated in terms of alkaline phosphatase activity.


Assuntos
Materiais Biomiméticos/química , Substitutos Ósseos/química , Colágeno/química , Engenharia Tecidual , Alicerces Teciduais/química , Animais , Materiais Biomiméticos/síntese química , Substitutos Ósseos/síntese química , Contagem de Células , Células Cultivadas , Reagentes de Ligações Cruzadas/síntese química , Reagentes de Ligações Cruzadas/química , Humanos , Células-Tronco Mesenquimais/citologia , Tamanho da Partícula , Porosidade , Salmão , Propriedades de Superfície
8.
Materials (Basel) ; 14(22)2021 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-34832488

RESUMO

The use of autologous bone graft for oral rehabilitation of bone atrophy is considered the gold standard. However, the available grafts do not allow a fast loading of dental implants, as they require a long healing time before full functionality. Innovative bioactive materials provide an easy-to-use solution to this problem. The current study shows the feasibility of calcium phosphate cement paste (Paste-CPC) in the sinus. Long implants were placed simultaneously with the cement paste, and provisional prosthetics were also mounted in the same sessions. Final prosthetics and the full loading took place within the same week. Furthermore, the study shows for the first time the possibility to monitor not only healing progression using Cone Beam Computer tomography (CBCT) but also material retention, over two years, on a case study example. The segmented images showed a 30% reduction of the cement size and an increased mineralized tissue in the sinus. Mechanical testing was performed qualitatively using reverse torque after insertion and cement solidification to indicate clinical feasibility. Both functional and esthetic satisfaction remain unchanged after one year. This flowable paste encourages the augmentation procedure with less invasive measure through socket of removed implants. However, this limitation can be addressed in future studies.

9.
Materials (Basel) ; 14(14)2021 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-34300793

RESUMO

Oil-based calcium phosphate cement (Paste-CPC) shows not only prolonged shelf life and injection times, but also improved cohesion and reproducibility during application, while retaining the advantages of fast setting, mechanical strength, and biocompatibility. In addition, poly(L-lactide-co-glycolide) (PLGA) fiber reinforcement may decrease the risk for local extrusion. Bone defects (diameter 5 mm; depth 15 mm) generated ex vivo in lumbar (L) spines of female Merino sheep (2-4 years) were augmented using: (i) water-based CPC with 10% PLGA fiber reinforcement (L3); (ii) Paste-CPC (L4); or (iii) clinically established polymethylmethacrylate (PMMA) bone cement (L5). Untouched (L1) and empty vertebrae (L2) served as controls. Cement performance was analyzed using micro-computed tomography, histology, and biomechanical testing. Extrusion was comparable for Paste-CPC(-PLGA) and PMMA, but significantly lower for CPC + PLGA. Compressive strength and Young's modulus were similar for Paste-CPC and PMMA, but significantly higher compared to those for empty defects and/or CPC + PLGA. Expectedly, all experimental groups showed significantly or numerically lower compressive strength and Young's modulus than those of untouched controls. Ready-to-use Paste-CPC demonstrates a performance similar to that of PMMA, but improved biomechanics compared to those of water-based CPC + PLGA, expanding the therapeutic arsenal for bone defects. O, significantly lower extrusion of CPC + PLGA fibers into adjacent lumbar spongiosa may help to reduce the risk of local extrusion in spinal surgery.

10.
Biomacromolecules ; 10(5): 1305-10, 2009 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-19344120

RESUMO

Textile chitosan fiber scaffolds were developed and tested in terms of biocompatibility for human bone marrow stromal cells (hBMSCs). A part of the scaffolds was further modified by coating with fibrillar collagen type I in order to biologize the surface. hBMSCs of two donors were used for cell culture experiments in vitro. Confocal laser scanning microscopy (CLSM) as well as scanning electron microscopy (SEM) revealed fast attachment and morphological adaptation of the cells on both the raw chitosan fibers and the collagen-coated scaffolds. Cells were osteogenically induced after 3 days and cultivated for up to 28 days on the scaffolds. Activity of lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) was analyzed to evaluate proliferation as well as osteogenic differentiation. We found a 3.5-6-fold increase in the cell number, whereas the collagen coating did not noticeably influence these factors. Osteogenic differentiation was confirmed by the course of ALP activity and immunostaining of osteocalcin. The feature of the collagen-coated as well as the raw chitosan fiber scaffolds to support attachment, proliferation, and differentiation of hBMSCs suggests a potential application of chitosan fibers and textile chitosan scaffolds for the tissue engineering of bone.


Assuntos
Materiais Biocompatíveis/química , Células da Medula Óssea/citologia , Quitosana/química , Células Estromais/citologia , Têxteis , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Fosfatase Alcalina/metabolismo , Contagem de Células , Técnicas de Cultura de Células , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Humanos , L-Lactato Desidrogenase/metabolismo , Teste de Materiais , Tamanho da Partícula , Propriedades de Superfície
11.
Drug Dev Ind Pharm ; 35(9): 1035-42, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19365781

RESUMO

BACKGROUND: Transdermal administration of estradiol offers advantages over oral estrogens for hormone replacement therapy regarding side effects by bypassing the hepatic presystemic metabolism. AIM: The objective of this study was to develop nanoparticles of Chondrosia reniformis sponge collagen as penetration enhancers for the transdermal drug delivery of 17beta-estradiol-hemihydrate in hormone replacement therapy. METHOD: Collagen nanoparticles were prepared by controlled alkaline hydrolysis and characterized using atomic force microscopy and photon correlation spectroscopy. Estradiol-hemihydrate was loaded to the nanoparticles by adsorption to their surface, whereupon a drug loading up to 13.1% of sponge collagen particle mass was found. After incorporation of drug-loaded nanoparticles in a hydrogel, the estradiol transdermal delivery from the gel was compared with that from a commercial gel that did not contain nanoparticles. RESULTS: Saliva samples in postmenopausal patients showed significantly higher estradiol levels after application of the gel with nanoparticles. The area under the curve (AUC) for estradiol time-concentration curves over 24 hours was 2.3- to 3.4-fold higher and estradiol levels 24 hours after administration of estradiol were at least twofold higher with the nanoparticle gel. CONCLUSIONS: The hydrogel with estradiol-loaded collagen nanoparticles enabled a prolonged estradiol release compared to a commercial gel and yielded a considerably enhanced estradiol absorption. Consequently, sponge collagen nanoparticles represent promising carriers for transdermal drug delivery.


Assuntos
Colágeno/química , Estradiol/administração & dosagem , Estradiol/farmacocinética , Poríferos/química , Absorciometria de Fóton , Administração Cutânea , Idoso , Animais , Disponibilidade Biológica , Diálise , Portadores de Fármacos , Eletroquímica , Excipientes , Feminino , Humanos , Hidrogéis , Microscopia de Força Atômica , Pessoa de Meia-Idade , Nanopartículas , Tamanho da Partícula , Saliva/metabolismo , Termodinâmica
12.
Drug Dev Ind Pharm ; 35(11): 1384-8, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19832639

RESUMO

BACKGROUND: Enteric coating prevents oral dose forms from being digested in the stomach, which is required for drugs that are acid unstable, have an irritant effect on the stomach, or are designed to act in the small intestine. AIM: The objective of this study was to develop a novel gastroresistant delayed-release tablet coating based on the marine sponge Chondrosia reniformis Nardo and to investigate the technical feasibility of the coating process. METHOD: An aqueous gastroresistant coating dispersion on the base of freeze-dried sponge collagen 15% (w/w) as the film-forming agent was developed. The disintegration test for gastroresistant tablets (Ph. Eur.) was carried out at increasing coating levels to reveal the required collagen layer thickness. Reproducibility of the method, physical properties, and stability of the coated tablets were investigated. RESULTS: Tablets coated with 13 mg/cm(2) of sponge collagen resisted more than 2 hours to 0.1 M hydrochloric acid, and disintegration of all tablets occurred within 10 minutes in phosphate buffer solution (pH 6.8). The method was reproducible, the mechanical properties of the coated tablets were satisfactory, and the obtained tablets could be stored for at least 6 months without loosing enteric properties. CONCLUSIONS: The novel coating based on the marine sponge collagen (using 12.9 mg/cm(2) coating material) complied with the requirements of Ph. Eur. for gastroresistant tablets. This coating material also meets the regulatory requirements for dietary supplements.


Assuntos
Colágeno/química , Portadores de Fármacos/química , Poríferos/química , Comprimidos com Revestimento Entérico/química , Animais , Colágeno/isolamento & purificação , Portadores de Fármacos/isolamento & purificação , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Solubilidade , Propriedades de Superfície
13.
Biomed Mater ; 14(3): 035015, 2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30870824

RESUMO

Isolated nanospheres consisting of organically modified hydroxyapatite (ormoHAP), prepared by an electric field-assisted ion double migration process, were embedded in foamed gelatin to form a composite scaffold. Degradation rates have been demonstrated to correlate with the crosslinking degree (40%, 80%) as well as with the mineral content of the scaffolds (0%, 20%, 40%). A human co-culture model of osteoblasts and osteoclasts, derived from bone marrow stromal cells and monocytes, respectively, without external addition of the factors RANKL and M-CSF, was run for up to 42 d in order to characterize the action of the ormoHAP-gelatin scaffolds on the co-culture. Examination was performed by quantitative biochemical methods (DNA, LDH, ALP, TRAP5b), gene expression analysis (ALP, BSP II, RANKL, IL-6, VTNR, CTSK, TRAP, OSCAR, CALCR) and confocal laser scanning microscopy (cell nuclei, actin, CD68, TRAP). Results confirm that ormoHAP embedded in the gelatin matrix enhanced TRAP 5b activity. As a feedback, ALP activity and gene expression of BSP II of osteoblasts increased. Finally, a sequence of cell cross-talk actions is suggested, which can explain the behavior of the formed vital co-culture and moreover the influence of the presence and concentration of ormoHAP.


Assuntos
Durapatita/química , Nanosferas/química , Osteoblastos/citologia , Osteoclastos/citologia , Fosfatase Alcalina/metabolismo , Células da Medula Óssea/citologia , Diferenciação Celular , Técnicas de Cocultura , DNA/química , Gelatina/química , Perfilação da Expressão Gênica , Humanos , Processamento de Imagem Assistida por Computador , Íons , Células-Tronco Mesenquimais/citologia , Monócitos/citologia , Osteócitos/citologia , Alicerces Teciduais
14.
Materials (Basel) ; 12(21)2019 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-31652704

RESUMO

Bone graft substitutes in orthopedic applications have to fulfill various demanding requirements. Most calcium phosphate (CaP) bone graft substitutes are highly porous to achieve bone regeneration, but typically lack mechanical stability. This study presents a novel approach, in which a scaffold structure with appropriate properties for bone regeneration emerges from the space between specifically shaped granules. The granule types were tetrapods (TEPO) and pyramids (PYRA), which were compared to porous CaP granules (CALC) and morselized bone chips (BC). Bulk materials of the granules were mechanically loaded with a peak pressure of 4 MP; i.e., comparable to the load occurring behind an acetabular cup. Mechanical loading reduced the volume of CALC and BC considerably (89% and 85%, respectively), indicating a collapse of the macroporous structure. Volumes of TEPO and PYRA remained almost constant (94% and 98%, respectively). After loading, the porosity was highest for BC (46%), lowest for CALC (25%) and comparable for TEPO and PYRA (37%). The pore spaces of TEPO and PYRA were highly interconnected in a way that a virtual object with a diameter of 150 µm could access 34% of the TEPO volume and 36% of the PYRA volume. This study shows that a bulk of dense CaP granules in form of tetrapods and pyramids can create a scaffold structure with load capacities suitable for the regeneration of an acetabular bone defect.

15.
Biomacromolecules ; 9(10): 2913-20, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18771318

RESUMO

Two novel scaffold models made of chitosan fibers were designed, fabricated, and investigated. Raw chitosan fibers were either tightened between plastic rings or were processed into stand-alone scaffolds. Chitosan fiber scaffolds were further modified by coating with a thin layer of fibrillar collagen type I to biologize the surface. Cell culture experiments were carried out using murine osteoblast-like cells (7F2). Confocal laser scanning microscopy (cLSM) as well as scanning electron microscopy (SEM) revealed fast attachment and morphological adaptation of the cells on both the raw chitosan fibers and the collagen-coated scaffolds. Cells were cultivated for up to 4 weeks on the materials and proliferation as well as osteogenic differentiation was quantitatively analyzed in terms of lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) activity. We found a 14-16-fold increase of cell number and the typical pattern of ALP activity, whereas the collagen coating does not remarkably influence these parameters. The maintenance of osteogenic phenotype on the novel materials was furthermore confirmed by immunostaining of osteocalcin and study of matrix mineralization. The feature of the collagen-coated but also the raw chitosan fiber scaffolds to support the attachment, proliferation, and differentiation of osteoblast-like cells suggest a potential application of chitosan fibers and textile chitosan scaffolds for the tissue engineering of bone.


Assuntos
Materiais Biocompatíveis/química , Quitosana/química , Osteoblastos/metabolismo , Têxteis , Fosfatase Alcalina/metabolismo , Animais , Cálcio/metabolismo , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Colorimetria/métodos , L-Lactato Desidrogenase/metabolismo , Camundongos , Microscopia Confocal , Microscopia Eletrônica de Varredura
16.
J Biomed Mater Res B Appl Biomater ; 106(3): 1165-1173, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-28556562

RESUMO

The ability of silica-/collagen-based composite xerogels to act as drug delivery systems was evaluated by taking into account the initial drug concentration, bioactivity of the xerogels, liquid, and incubation regime. The proteasome inhibitor bortezomib was chosen as a model drug, used for the systemic treatment of multiple myeloma. Incubation during 14 days in phosphate-buffered saline (PBS) or simulated body fluid (SBF) showed a weak initial burst and was identified to be of first order with subsequent release being independent from the initial load of 0.1 or 0.2 mg bortezomib per 60 mg monolithic sample. Faster drug release occurred during incubation in SBF compared to PBS, and during static incubation without changing the liquid, compared to dynamic incubation with daily liquid changes. Drug-loaded xerogels with hydroxyapatite as a third component exhibited enhanced bioactivity retarding drug release, explained by formation of a surface calcium phosphate layer. The fastest release of 50% of the total drug load was observed for biphasic xerogels after 7 days during dynamic incubation in SBF. As a result, the presented concept is suitable for the intended combination of the advantageous bone substitution properties of xerogels and local application of drugs exemplified by bortezomib. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1165-1173, 2018.


Assuntos
Antineoplásicos/administração & dosagem , Antineoplásicos/farmacocinética , Bortezomib/administração & dosagem , Bortezomib/farmacocinética , Materiais Biocompatíveis , Líquidos Corporais/química , Cimentos Ósseos , Substitutos Ósseos , Fosfatos de Cálcio , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Durapatita , Géis , Microscopia Eletrônica de Varredura , Dióxido de Silício
17.
J Orthop Res ; 36(1): 106-117, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28574614

RESUMO

Calcium phosphate cements (CPCs) are widely used for bone-defect treatment. Current developments comprise the fabrication of porous scaffolds by three-dimensional plotting and doting using biologically active substances, such as strontium. Strontium is known to increase osteoblast activity and simultaneously to decrease osteoclast resorption. This study investigated the short- and long-term in vivo performances of strontium(II)-doted CPC (SrCPC) scaffolds compared to non-doted CPC scaffolds after implantation in unloaded or load-bearing trabecular bone defects in sheep. After 6 weeks, both CPC and SrCPC scaffolds exhibited good biocompatibility and osseointegration. Fluorochrome labeling revealed that both scaffolds were penetrated by newly formed bone already after 4 weeks. Neither strontium doting nor mechanical loading significantly influenced early bone formation. In contrast, after 6 months, bone formation was significantly enhanced in SrCPC compared to CPC scaffolds. Energy dispersive X-ray analysis demonstrated the release of strontium from the SrCPC into the bone. Strontium addition did not significantly influence material resorption or osteoclast formation. Mechanical loading significantly stimulated bone formation in both CPC and SrCPC scaffolds after 6 months without impairing scaffold integrity. The most bone was found in SrCPC scaffolds under load-bearing conditions. Concluding, these results demonstrate that strontium doting and mechanical loading additively stimulated bone formation in CPC scaffolds and that the scaffolds exhibited mechanical stability under moderate load, implying good clinical suitability. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:106-117, 2018.


Assuntos
Cimentos Ósseos , Fosfatos de Cálcio/química , Osteogênese/efeitos dos fármacos , Estrôncio/farmacologia , Alicerces Teciduais , Animais , Feminino , Osseointegração , Ovinos , Estresse Mecânico
18.
J Biomed Mater Res B Appl Biomater ; 106(7): 2565-2575, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29316208

RESUMO

The combination of the two techniques of rapid prototyping 3D-plotting and bioactive surface functionalization is presented, with emphasis on the in vitro effect of Bone Sialoprotein (BSP) on primary human osteoblasts (hOBs). Our primary objective was to demonstrate the BSP influence on the expression of distinctive osteoblast markers in hOBs. Secondary objectives included examinations of the scaffolds' surface and the stability of BSP-coating as well as investigations of cell viability and proliferation. 3D-plotted calcium phosphate cement (CPC) scaffolds were coated with BSP via physisorption. hOBs were seeded on the coated scaffolds, followed by cell viability measurements, gene expression analysis and visualization. Physisorption is an effective method for BSP-coating. Coating with higher BSP concentrations leads to enhanced BSP release. Two BSP concentrations (50 and 200 µg/mL) were examined in this study. The lower BSP concentration (50 µg/mL) decreased ALP and SPARC expression, whereas the higher BSP concentration (200 µg/mL) did not change gene marker expression. Enhanced cell viability was observed on BSP-coated scaffolds on day 3. hOBs developed a polygonal shape and connected in an intercellular network under BSP influence. Quantitative cell morphology analyses demonstrated for BSP-coated CPCs an enhanced cell area and reduced circularity. The strength of the above-mentioned effects of BSP-coated scaffolds in vivo is unknown, and future work is focusing on bone ingrowth and vascularization in vivo. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2565-2575, 2018.


Assuntos
Fosfatos de Cálcio/química , Materiais Revestidos Biocompatíveis/química , Sialoproteína de Ligação à Integrina/química , Osteoblastos/metabolismo , Impressão Tridimensional , Alicerces Teciduais/química , Humanos , Teste de Materiais , Osteoblastos/citologia
19.
Injury ; 47 Suppl 1: S52-61, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26768293

RESUMO

New bone formation was studied in a metaphyseal fracture-defect in ovariectomized rats stimulated by a plain and a strontium-enriched macroporous silica/collagen scaffold (ScB30 and ScB30Sr20) and a compact silica/collagen xerogel (B30). 45 female Sprague-Dawley rats were randomly assigned to three different treatment groups: (1) ScB30 (n=15), (2) ScB30Sr20 (n=15), and (3) B30 (n=15). 12 weeks after bilateral ovariectomy and multi-deficient diet, a 4 mm wedge-shaped fracture-defect was created at the metaphyseal area of the left femur. A 7-hole T-shaped plate at the lateral aspect of the femur stabilized the bone and the defect was filled with ScB30, ScB30Sr20 or B30 subsequently. After six weeks, histomorphometrical analysis revealed a statistically significant higher bone volume/tissue volume ratio in the ScB30Sr20 group compared to ScB30 (p=0.043) and B30 (p=0.0001) indicating an improved formation of new bone by the strontium-enriched macroporous silica/collagen scaffold. Furthermore, immunohistochemical results showed increased expression of BMP2 and OPG and a decreased RANKL expression in the ScB30Sr20 group. This was further confirmed with the gene expression analysis where an increase in prominent bone formation markers (ALP, OCN, Runx2, Col1a1 and Col10a1) was seen. No material remnants were found in the scaffold group indicating an almost complete degradation process of the biomaterials. This is confirmed by ToF-SIMS analysis that did not detect any strontium in the ScB30Sr20 group neither in the defect nor in the surrounding tissue. Taken together, this study shows the stimulating effects of strontium through increased bone formation by up regulation of osteoanabolic markers. This work also indicates the importance of material porosity, geometry and biodegradability in bone healing.


Assuntos
Materiais Biocompatíveis/farmacologia , Receptores de Proteínas Morfogenéticas Ósseas Tipo II/metabolismo , Estrôncio/farmacologia , Fraturas da Tíbia/patologia , Animais , Cimentos Ósseos/farmacologia , Fosfatos de Cálcio/farmacologia , Feminino , Osteogênese/efeitos dos fármacos , Ovariectomia , Ratos , Ratos Sprague-Dawley
20.
J Biomed Mater Res A ; 103(2): 525-33, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24737729

RESUMO

Lactoferrin is a milk-derived glycoprotein with anabolic effects on the bone tissue. In this study, artificial extracellular matrices (aECM) consisting of collagen type I fibrils formed in the presence of lactoferrin at two different concentrations (0.5 and 1 mg mL(-1) ) were prepared on the surface of poly(lactic-co-glycolic acid) (PLGA) foils. The aim of the study was to investigate the effects of aECM on the adhesion, growth and osteogenic differentiation of human osteoblast-like Saos-2 cells. On days 1 and 3 after seeding, higher numbers of cells were found on samples with collagen and collagen-lactoferrin coatings (particularly on those formed at the higher concentration of lacroferrin) than on control microscopic glass coverslips. Cells on coatings formed in the presence of lactoferrin had more numerous and better developed vinculin-containing focal adhesion plaques. On day 7, cells on coatings with and without lactoferrin produced significantly higher levels of osteocalcin than cells on control polystyrene cell culture dishes, the highest average values being found on samples with the lower concentration of lactoferrin. Expression of collagen I and alkaline phosphatase was on a similar level in cells on all tested samples and control polystyrene. Thus, lactoferrin promotes adhesion, growth and osteogenic differentiation of Saos-2 cells and is promising as a bone implant coating component.


Assuntos
Diferenciação Celular , Proliferação de Células , Materiais Revestidos Biocompatíveis/química , Colágeno/química , Lactoferrina/química , Osteoblastos/metabolismo , Peptídeos Catiônicos Antimicrobianos/química , Linhagem Celular , Humanos , Osteoblastos/citologia
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