ABSTRACT
Trauma or disease inflicted by tissue injuries may cause tissue degeneration. The use of biomaterials for direct or indirect repair has emerged as a promising alternative, and has become an important research topic. The pequi fruit (Caryocar brasiliense Camb.) has shown antifungal, antibacterial, anti-inflammatory, healing, antitumor, and antioxidant properties. The objective of this study was to develop a new biomaterial using a combination of collagen, gelatin, and pulp pequi oil, and to evaluate its biocompatibility in comparison with that of biomaterials produced without pulp pequi oil. Membranes were prepared from a mixture of bovine tendon collagen, commercial gelatin, and pulp pequi oil. The inflammatory and cicatricial processes were assessed via histopathology of the tissue interface/implants in the subcutaneous tissues and quantitative evaluation of leukocyte and collagen production in Wistar rats. It was observed that the presence of pequi oil reduced the amount of foreign-body giant cells and favored the recruitment of fibroblasts (P< 0.01), thereby promoting greater production of collagen membrane than that in the membranes of control samples. Therefore, it can be concluded that the addition of pequi oil improved the biocompatibility of collagen and accelerated the healing process.(AU)
Trauma ou lesões causadas por doenças podem enfraquecer e degenerar os tecidos humanos e animais. O uso de biomateriais para reparação direta ou indireta surgiu como uma alternativa promissora e tornou-se um importante tema de pesquisa. O óleo de pequi (Caryocar brasiliense Camb.) mostrou propriedades antifúngicas, antibacterianas, anti-inflamatórias, curativas, antitumorais e antioxidantes. O objetivo deste estudo foi obter um novo biomaterial, produzido pela combinação de óleo de pequi, colágeno e gelatina, para avaliar sua biocompatibilidade em comparação às membranas produzidas sem o óleo. As membranas foram preparadas por meio da mistura de colágeno de tendão bovino, gelatina comercial e óleo de pequi. Os processos inflamatórios e cicatriciais foram avaliados por histopatologia da interface / implantes de tecido subcutâneo de ratos Wistar para avaliação quantitativa da produção de leucócitos e colágeno. Observou-se que a presença de óleo de pequi reduziu a quantidade de células gigantes de corpo estranho e favoreceu o recrutamento de fibroblastos (P<0,01), promovendo, assim, maior produção da membrana de colágeno em comparação com a membrana de controle. Portanto, pode-se concluir que a adição de óleo de pequi melhorou a biocompatibilidade do colágeno e acelerou o processo de cicatrização.(AU)
Subject(s)
Animals , Rats , Biocompatible Materials/therapeutic use , Soft Tissue Injuries/veterinary , Ericales , Wound Healing , Collagen/therapeutic use , Gelatin/therapeutic useABSTRACT
In traumatology, we encounter several clinical challenges that involve extensive bone loss primarily related to trauma, conditions that can be treated with autologous grafts. A good alternative is the use of synthetic biomaterials as substitutes. These polymers provide a suitable environment for the growth of new bone and vascular tissue, which are essential for repair. Collagen/hydroxyapatite composites have proven to be biocompatible and to behave mechanically. Furthermore, the addition of chitosan contributes to the formation of a three-dimensional structure that permits cell adhesion and proliferation, thus improving osteogenesis. The aim of this study was to evaluate bone formation during the repair of bone defects experimentally induced in the skull of rats and grafted with a polymer blend consisting of bovine tendon collagen and chitosan combined with hydroxyapatite. Thirty animals were used for the creation of a defect in the left parietal bone and were divided into three groups of 10 animals each: a control group without biomaterial implantation, a group receiving the blend of collagen and chitosan, and a group receiving this blend combined with hydroxyapatite. Each group was subdivided and the animals were sacrificed 3 or 8 weeks after surgery. After sacrifice, the skulls were removed for macroscopic photodocumentation and radiographic examination. The samples were processed for histological evaluation of new bone formation at the surgical site. Macroscopic and radiographic analysis demonstrated the biocompatibility of the blends. Histologically, the formation of new bone occurred in continuity with the edges of the defect, with the observation of higher volumes in the grafted groups compared to control. Mineralization of sponges did not stimulate bone neoformation, with bone repair being incomplete over the experimental period. In conclusion, mineralization by the addition of hydroxyapatite should be better studied. However, the collagen/chitosan sponges used in this study are suitable to stimulate osteogenesis in cranial defects, although this process is slow and not sufficient to achieve complete bone regeneration over a short period of time.
Subject(s)
Biocompatible Materials/chemistry , Bone Substitutes/chemistry , Chitosan/pharmacology , Collagen/pharmacology , Durapatite/pharmacology , Osteogenesis/physiology , Skull Fractures/pathology , Animals , Bone Morphogenetic Protein 2 , Cell Adhesion , Cell Proliferation , Disease Models, Animal , Male , Rats , Rats, Wistar , Skull Fractures/surgeryABSTRACT
The main purpose of the present work was to evaluate if low laser level therapy (LLLT) can improve the effects of Biosilicate®/PLGA (BS/PLGA) composites on cell viability and bone consolidation using a tibial defects of rats. The composites were characterized by scanning electron microscope (SEM) and reflection Fourier transform infrared spectrometer (FTIR). For the in vitro study, fibroblast and osteoblast cells were seeded in the extract of the composites irradiated or not with LLLT (Ga-Al-As, 808nm, 10J/cm2) to assess cell viability after 24, 48 and 72h. For the in vivo study, 80 Wistar rats with tibial bone defects were distributed into 4 groups (BS; BS+LLLT; BS/PLGA and BS/PLGA+LLLT) and euthanized after 2 and 6weeks. Laser irradiation Ga-Al-As (808nm, 30J/cm2) in the rats was performed 3 times a week. The SEM and FTIR results revealed that PLGA were successfully inserted into BS and the microparticles degraded over time. The in vitro findings demonstrated higher fibroblast viability in both BS/PLGA groups after 24h and higher osteoblast viability in BS/PLGA+LLLT in all periods. As a conclusion, animals treated with BS/PLGA+LLLT demonstrated an improved material degradation and an increased amount of granulation tissue and newly formed bone.
Subject(s)
Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Lactic Acid/chemistry , Low-Level Light Therapy , Osteogenesis/drug effects , Osteogenesis/radiation effects , Polyglycolic Acid/chemistry , Silicates/chemistry , Animals , Biomechanical Phenomena , Cell Line , Cell Survival/drug effects , Cell Survival/radiation effects , Male , Polylactic Acid-Polyglycolic Acid Copolymer , Rats , Rats, WistarABSTRACT
Studies indicate that hyperthermic therapy using gold nanorods and photodynamic activity with many photosensitizers can present a synergistic effect, and offer a great therapeutic potential, although more investigation needs to be performed before such approach could be implemented. We proposed to investigate the effect of the attachment of phthalocyanines on the surface of gold nanorods (well-characterized devices for hyperthermia generation) for the elimination of melanoma, one of the most important skin cancers due to its high lethality. Following the synthesis of nanorods through a seed-mediated method, the efficacy of photodynamic therapy (PDT) and hyperthermia was assessed separately. We chose to coat the nanorods with two tetracarboxylated zinc phthalocyanines - with or without methyl-glucamine groups. After the coating process, the phthalocyanines formed ionic complexes with the cetyltrimethylammonium bromide (CTAB) that was previously covering the nanoparticles. The nanorod-phthalocyanines complexes were analyzed by transmission electron microscopy (TEM), and their singlet oxygen and hydroxyl radical generation yields were assessed. Furthermore, they were tested in vitro with melanotic B16F10 and amelanotic B16G4F melanoma cells. The cells with nanoparticles were irradiated with laser (at 635nm), and the cell viability was assessed. The results indicate that the photodynamic properties of the phthalocyanines tested are enhanced when they are attached on the nanorods surface, and the combination of PDT and hyperthermia was able to eliminate over 90% of melanoma cells. This is a novel study because two tetracarboxylated phthalocyanines were used and because the same wavelength was irradiated to activate both the nanorods and the photosensitizers.
Subject(s)
Gold/chemistry , Indoles/chemistry , Nanotubes/chemistry , Organometallic Compounds/chemistry , Photosensitizing Agents/chemistry , Animals , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Proliferation/radiation effects , Cell Survival/drug effects , Cell Survival/radiation effects , Humans , Hydroxyl Radical/metabolism , Isoindoles , Lasers , Melanoma, Experimental/drug therapy , Mice , Microscopy, Electron, Transmission , Photochemotherapy , Photosensitizing Agents/pharmacology , Photosensitizing Agents/therapeutic use , Singlet Oxygen/metabolism , Zinc CompoundsABSTRACT
The aims of this study were to characterize different BS/PLGA composites for their physicochemical and morphological characteristics and evaluate the in vitro and in vivo biological performance. The physicochemical and morphological modifications were analyzed by pH, mass loss, XRD, setting time, and SEM. For in vitro analysis, the osteoblast and fibroblast viability was evaluated. For in vivo evaluations, histopathology and immunohistochemistry were performed in a tibial defect in rats. After incubation, all composites presented lower values in pH and mass loss over time. Moreover, XRD and SEM analysis confirmed that the composites degraded over time. Additionally, pore formation was observed by SEM analysis after incubation mainly in BS/PLGA groups. BS/PLGA showed significantly increased in osteoblast viability 24 h. Moreover, BS/PLGA composites demonstrated an increase in fibroblast viability in all periods analyzed when compared to BS. In the in vivo study, after 2 and 6 weeks of implantation of biomaterials, histopathological findings revealed that the BS/PLGA composites degrades over time, mainly at periphery. Moreover, can be observed the presence of granulation tissue, bone formation, Runx-2, and RANKL immunoexpression in all groups. In conclusion, BS/PLGA composites present appropriate physicochemical characteristics, stimulate the cellular viability, and enhance the bone repair in vivo. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1063-1074, 2017.
Subject(s)
Lactic Acid , Materials Testing , Osteoblasts/metabolism , Polyglycolic Acid , Silicates , Tibia/metabolism , Tibial Fractures/therapy , Animals , Cell Line , Cell Survival/drug effects , Fibroblasts/cytology , Fibroblasts/pathology , Hydrogen-Ion Concentration , Lactic Acid/chemistry , Lactic Acid/pharmacology , Mice , Osteoblasts/pathology , Osteogenesis/drug effects , Polyglycolic Acid/chemistry , Polyglycolic Acid/pharmacology , Polylactic Acid-Polyglycolic Acid Copolymer , Rats , Silicates/chemistry , Silicates/pharmacology , Tibia/pathology , Tibial Fractures/metabolism , Tibial Fractures/pathologyABSTRACT
As lesões em membros de grandes animais são um desafio para médicos veterinários, uma vez que somente a osteossíntese não garante resultados satisfatórios. Muitos pesquisadores vêm se dedicando ao desenvolvimento e estudo de substitutos ósseos produzidos de materiais naturais, como quitosana, colágeno e hidroxiapatita, que auxiliam na regeneração óssea. Seis ovinos fêmeas da raça Santa Inês foram submetidos a ostectomias unicorticais de sete milímetros de diâmetro na região proximal da superfície dorsomedial dos III/IV metacarpianos. Foi implantado compósito de quitosana, colágeno e hidroxiapatita em um membro torácico para avaliação da biocompatibilidade do material ao tecido ósseo ovino, e no membro contralateral foi reproduzida a mesma técnica, porém foi mantido sem preenchimento, como controle. Após 60 dias do procedimento cirúrgico, realizou-se biópsia óssea na área de interface entre biomaterial/osso (membro com compósito) e tecido neoformado/osso (membro controle), para realização de avaliação histológica do material não descalcificado, por meio de microscopia de luz e microscopia eletrônica de varredura. Na análise histomorfométrica, mediante microscopia de luz, foi possível identificar maior porcentagem de tecido neoformado em membro controle, quando comparado ao membro com compósito (80% e 63,5%, respectivamente; P<0,05). Por meio da microscopia eletrônica de varredura, observou-se invasão da estrutura interna do compósito por tecido ósseo neoformado. Não houve formação de tecido cicatricial, reação de corpo estranho ou resposta inflamatória crônica nas amostras analisadas. Conclui-se que o compósito de quitosana, colágeno e hidroxiapatita, quando implantado em tecido ósseo ovino, apresenta biocompatibilidade e perfil osteocondutor.(AU)
Fracture management poses a great challenge to large animal practitioners. Osteosynthesis alone is often insufficient to provide satisfactory outcomes in large animals; therefore, several research efforts have been made to investigate and develop bone substitutes capable of promoting bone regeneration. Chitosan-collagen-hydroxyapatite composites constitute a promising alternative given their similar composition to bone. Six Santa Inês ewes were submitted to the creation of experimental 7mm wide unicortical defects on the dorsomedial aspect of the proximal III/IV metacarpal bone diaphysis. Limbs were randomly selected for treatment with chitosan-collagen-hydroxyapatite composite or to serve as untreated controls. Biopsy fragments were collected from the bone/new bone or the bone/biomaterial interface (control and treated defects respectively) within 60 days of surgery; composite biocompatibility was assessed using light and scanning electron microscopy. Histomorphometric analysis under light microscopy revealed greater percentage of new bone tissue in control compared to treated defects (80% and 63.5% respectively; P<0.05). No scar tissue formation, foreign body or chronic inflammatory reactions were observed. Scanning electron microscopy revealed invasion of the composite by new bone tissue. The chitosan-collagen-hydroxyapatite composite studied is biocompatible with bone and shows osteoconductive properties in sheep.
Subject(s)
Animals , Biocompatible Materials/analysis , Bone and Bones , Bone Substitutes/analysis , Sheep , Bone Regeneration , Chitosan/therapeutic use , Collagen/therapeutic use , Hydroxyapatites , Microscopy, Electron, Scanning/veterinaryABSTRACT
The indication of biomaterials has increased substantially in the regenerative therapy of bone defects. However, in addition to evaluating the physicochemical properties of biomaterials, the quality of the recipient tissue is also essential for the osseointegration of implants, as abnormalities in bone metabolism, such as gonadal hormone deficiency, can influence bone healing. This study evaluated the osteoregenerative capacity of collagen membranes derived from bovine pericardium and intestinal serosa in the repair of cranial defects in ovariectomised rats. Thirty female Wistar rats were submitted to surgical creation of a 5-mm cranial bone defect. The rats were divided into a control group (not ovariectomised) and an ovariectomised group. The non-ovariectomised group was divided into three subgroups: control (G1) in which the defect was not filled with the biomaterial, and two subgroups (G2 and G3) that received the bovine pericardium- and serosa-derived collagen membranes, respectively. The ovariectomised group was divided into the same subgroups (G4, G5, and G6). The animals were sacrificed 8 weeks after surgery. The calvaria were removed for macroscopic and radiographic photodocumentation and processed for histomorphometric analysis of bone healing at the surgical site. Macroscopic, radiological, and microscopic analyses demonstrated the biocompatibility of the implanted collagen membranes, as indicated by the absence of infiltration and signs of inflammation at the surgical site. Histologically, discrete immature bone neoformation projecting from the margins of the defect was observed at the surgical site in ovariectomised groups when compared to the non-ovariectomised groups. The volume of newly formed bone was significantly higher in the non-ovariectomised groups (G1: 7.83%±1.32; G2: 21.33%±1.96; and G3: 22.83%±0.98) compared to the respective ovariectomised subgroups (G4: 3.16%±0.75; G5: 16.83%±0.98; and G6: 16.16%±0.75), thus demonstrating the deleterious effects of ovariectomy on bone homeostasis. Higher volumes of newly formed bone were observed in the groups receiving the membrane grafts (G2, G3, G5, and G6) compared to the control groups (G1 and G4). In conclusion, the bilateral ovariectomy compromises the ability to repair bone lesions grafted with osteoconductive biomaterials as in the case of collagen membranes derived from both bovine pericardium and intestinal serosa.
Subject(s)
Biocompatible Materials/pharmacology , Collagen/pharmacology , Pericardium/pathology , Skull/pathology , Wound Healing/physiology , Animals , Cattle , Disease Models, Animal , Feasibility Studies , Female , Ovariectomy , Rats , Rats, Wistar , Transplantation, HeterologousABSTRACT
This work describes the preparation and characterization of anionic collagen composites with rhamsan and vinylidene fluoride-trifluorethylene with improved rheological and dielectric properties without loss of collagen secondary structure with an interaction occurring between both macromolecules of the composites. On a comparative basis, the force needed for the extrusion of anionic collagen:rhamsan composites was in the range from 0.088 to 0.080 J compared to that for collagen of 0.189 J. Anionic collagen:vinylidene fluoride-trifluorethylene composites were characterized, in the case of the 1:1 composite, by a pyroelectric coefficient of 1.89 x 10(-4) cm(-2) K(-1), which was significantly higher than those determined under the same conditions for native anionic collagen and vinylidene fluoride-trifluorethylene.
