Use of Plant Extracts in Polymeric Scaffolds in the Regeneration of Mandibular Injuries.

Severe loss of bone mass may require grafting, and, among the alternatives available, there are natural biomaterials that can act as scaffolds for the cell growth necessary for tissue regeneration. Collagen and elastin polymers are a good alternative due to their biomimetic properties of bone tissue, and their characteristics can be improved with the addition of polysaccharides such as chitosan and bioactive compounds such as jatoba resin and pomegranate extract due to their antigenic actions. The aim of this experimental protocol was to evaluate bone neoformation in experimentally made defects in the mandible of rats using polymeric scaffolds with plant extracts added. Thirty rats were divided into group 1, with a mandibular defect filled with a clot from the lesion and no graft implant (G1-C, n = 10); group 2, filled with collagen/chitosan/jatoba resin scaffolds (G2-CCJ, n = 10); and group 3, with collagen/nanohydroxyapatite/elastin/pomegranate extract scaffolds (G3-CHER, n = 10). Six weeks after surgery, the animals were euthanized and samples from the surgical areas were submitted to macroscopic, radiological, histological, and morphometric analysis of the mandibular lesion repair process. The results showed no inflammatory infiltrates in the surgical area, indicating good acceptance of the scaffolds in the microenvironment of the host area. In the control group (G1), there was a predominance of reactive connective tissue, while in the grafted groups (G2 and G3), there was bone formation from the margins of the lesion, but it was still insufficient for total bone repair of the defect within the experimental period standardized in this study. The histomorphometric analysis showed that the mean percentage of bone volume formed in the surgical area of groups G1, G2, and G3 was 17.17 ± 2.68, 27.45 ± 1.65, and 34.07 ± 0.64 (mean ± standard deviation), respectively. It can be concluded that these scaffolds with plant extracts added can be a viable alternative for bone repair, as they are easily manipulated, have a low production cost, and stimulate the formation of new bone by osteoconduction.

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.

In Vivo Evaluation of Collagen and Chitosan Scaffold, Associated or Not with Stem Cells, in Bone Repair.

Natural polymers are increasingly being used in tissue engineering due to their ability to mimic the extracellular matrix and to act as a scaffold for cell growth, as well as their possible combination with other osteogenic factors, such as mesenchymal stem cells (MSCs) derived from dental pulp, in an attempt to enhance bone regeneration during the healing of a bone defect. Therefore, the aim of this study was to analyze the repair of mandibular defects filled with a new collagen/chitosan scaffold, seeded or not with MSCs derived from dental pulp. Twenty-eight rats were submitted to surgery for creation of a defect in the right mandibular ramus and divided into the following groups: G1 (control group; mandibular defect with clot); G2 (defect filled with dental pulp mesenchymal stem cells-DPSCs); G3 (defect filled with collagen/chitosan scaffold); and G4 (collagen/chitosan scaffold seeded with DPSCs). The analysis of the scaffold microstructure showed a homogenous material with an adequate percentage of porosity. Macroscopic and radiological examination of the defect area after 6 weeks post-surgery revealed the absence of complete repair, as well as absence of signs of infection, which could indicate rejection of the implants. Histomorphometric analysis of the mandibular defect area showed that bone formation occurred in a centripetal fashion, starting from the borders and progressing towards the center of the defect in all groups. Lower bone formation was observed in G1 when compared to the other groups and G2 exhibited greater osteoregenerative capacity, followed by G4 and G3. In conclusion, the scaffold used showed osteoconductivity, no foreign body reaction, malleability and ease of manipulation, but did not obtain promising results for association with DPSCs.

In Vivo Study of Nasal Bone Reconstruction with Collagen, Elastin and Chitosan Membranes in Abstainer and Alcoholic Rats.

The aim of the present study was to evaluate the use of collagen, elastin, or chitosan biomaterial for bone reconstruction in rats submitted or not to experimental alcoholism. Wistar male rats were divided into eight groups, submitted to chronic alcohol ingestion (G5 to G8) or not (G1 to G4). Nasal bone defects were filled with clot in animals of G1 and G5 and with collagen, elastin, and chitosan grafts in G2/G6, G3/G7, and G4/G8, respectively. Six weeks after, all specimens underwent radiographic, tomographic, and microscopic evaluations. Bone mineral density was lower in the defect area in alcoholic animals compared to the abstainer animals. Bone neoformation was greater in the abstainer groups receiving the elastin membrane and in abstainer and alcoholic rats receiving the chitosan membrane (15.78 ± 1.19, 27.81 ± 0.91, 47.29 ± 0.97, 42.69 ± 1.52, 13.81 ± 1.60, 18.59 ± 1.37, 16.54 ± 0.89, and 37.06 ± 1.17 in G1 to G8, respectively). In conclusion, osteogenesis and bone density were more expressive after the application of the elastin matrix in abstainer animals and of the chitosan matrix in both abstainer and alcoholic animals. Chronic alcohol ingestion resulted in lower bone formation and greater formation of fibrous connective tissue.

In Vitro Evaluation of Acellular Collagen Matrices Derived from Porcine Pericardium: Influence of the Sterilization Method on Its Biological Properties.

The aim of this study were characterize acellular collagen matrices derived from porcine pericardium (PP) and to evaluate their properties after sterilization by ethylene oxide and gamma ray. PP matrices were subjected to alkaline hydrolysis (AH), and samples were characterized for biological stability, membrane thickness measurements, differential scanning calorimetry (DSC) and scanning electron microscopy (SEM). Subsequently, the matrices were frozen, lyophilized and sterilized by ethylene oxide or gamma radiation. For in vitro assays, CHO-K1 cell culture was used and evaluated for cytotoxicity, clonogenic survival assay, genotoxicity and mutagenicity. Analysis of variance (ANOVA) was used, followed by Dunnett's post-test, with a significance level of 5%. After AH, there was no significant change in matrix thickness. The relative biodegradability of the material after implantation was observed. Morphology and dimensions had small changes after AH. As for cell viability, none of the tested matrices showed a statistically significant difference (p > 0.05; Dunnett) regardless of the sterilization method. Furthermore, it was found that PP matrices did not interfere with the proliferation capacity of CHO-K1 cells (p > 0.05; Dunnett). As for genotoxicity, when sterilized with ethylene oxide (NP, P12 and P24), it showed genotoxic potential, but it was not genotoxic when sterilized by gamma radiation. No mutagenic effects were observed in either group. PP-derived collagen matrices hydrolyzed at different times were not cytotoxic. It is concluded that the best method of sterilization is through gamma radiation, since no significant changes were observed in the properties of the PP matrices.

Chitosan/gelatin films with jatobá resin: Control of properties by vegetal resin inclusion and degree of acetylation modification.

Blending chitosan and gelatin, two biodegradable and non-toxic polymers, is a recurrent choice in food coating or biomaterials development. The incorporation of vegetal extracts into chitosan/gelatin films can improve or introduce some properties to these materials. Jatobá resin is a product of Hymenaea genus trees with antimicrobial and anti-inflammatory activities, interesting properties for films applied in several areas. The chitosan degree of acetylation (DA) influences the inter and intramolecular interactions of this polymer and, therefore, also implicates in changes of its properties. This research aims to study the influence of jatobá resin inclusion and chitosan DA modification on chitosan/gelatin films properties. Both jatobá resin and chitosan DA affected physicochemical, antimicrobial and barrier properties of the films, allowing the control of these properties by changes in these parameters. Jatobá resin incorporation and the decrease in chitosan DA significantly improved antimicrobial activity and water vapor permeability of films with the reduction of water solubility and swelling.

In Vivo Biological Behavior of Polymer Scaffolds of Natural Origin in the Bone Repair Process.

Autologous bone grafts, used mainly in extensive bone loss, are considered the gold standard treatment in regenerative medicine, but still have limitations mainly in relation to the amount of bone available, donor area, morbidity and creation of additional surgical area. This fact encourages tissue engineering in relation to the need to develop new biomaterials, from sources other than the individual himself. Therefore, the present study aimed to investigate the effects of an elastin and collagen matrix on the bone repair process in critical size defects in rat calvaria. The animals (Wistar rats, n = 30) were submitted to a surgical procedure to create the bone defect and were divided into three groups: Control Group (CG, n = 10), defects filled with blood clot; E24/37 Group (E24/37, n = 10), defects filled with bovine elastin matrix hydrolyzed for 24 h at 37 °C and C24/25 Group (C24/25, n = 10), defects filled with porcine collagen matrix hydrolyzed for 24 h at 25 °C. Macroscopic and radiographic analyses demonstrated the absence of inflammatory signs and infection. Microtomographical 2D and 3D images showed centripetal bone growth and restricted margins of the bone defect. Histologically, the images confirmed the pattern of bone deposition at the margins of the remaining bone and without complete closure by bone tissue. In the morphometric analysis, the groups E24/37 and C24/25 (13.68 ± 1.44; 53.20 ± 4.47, respectively) showed statistically significant differences in relation to the CG (5.86 ± 2.87). It was concluded that the matrices used as scaffolds are biocompatible and increase the formation of new bone in a critical size defect, with greater formation in the polymer derived from the intestinal serous layer of porcine origin (C24/25).

Development of collagen/nanohydroxyapatite scaffolds containing plant extract intended for bone regeneration.

In this study scaffolds of nanohydroxyapatite (nHA) and anionic collagen (C) combined with plant extracts intended for bone tissue repair were developed. Grape seed (P), pomegranate peel (R) and jabuticaba peel (J) extracts were used as collagen crosslinker agents in order to improve the materials properties. All crude extracts were effective against Staphylococcus aureus, but only for CR scaffold inhibition zone was noticed. The extracts acted as crosslinking agents, increasing enzymatic resistance and thermal stability of collagen. The extracts showed cytotoxicity at the concentrations tested, while nHA increased cell viability. The scaffolds presented porosity and pore size appropriate for bone growth. CR, CnHAP, CnHAR and CnHAJ increased the cell viability after 24 h. The combination of collagen, nHA and plant extracts offers a promising strategy to design novel biomaterials for bone tissue regeneration.

In vitro evaluation of hydroxyapatite, chitosan, and carbon nanotube composite biomaterial to support bone healing / Avaliação in vitro do biomaterial compósito à base de hidroxiapatita, quitosana e nanotubo de carbono como adjuvante no reparo ósseo

Hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were developed to improve bone healing. Previous studies suggested that a combination of biomaterials and mesenchymal stem cells (MSCs) can potentially help promote bone regeneration. In the present study, we first developed hydroxyapatite, chitosan, and carbon nanotube composite biomaterial. Then, the effect of different concentrations of the extract on the viability of Vero cells (ATCC CCL-81) and MSCs obtained from sheep bone marrow using methylthiazol tetrazolium (MTT) and propidium iodide (PI) assays were evaluated. The biomaterial group demonstrated an absence of cytotoxicity, similar to the control group. Samples with 50% and 10% biomaterial extract concentrations showed higher cell viability compared to samples from the control group (MTT assay). These results suggest that the presence of this composite biomaterial can be used with MSCs. This study also concluded that hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were not cytotoxic. Therefore, these could be used for performing in vivo tests.(AU)

O compósito à base de hidroxiapatita, quitosana e nanotubo de carbono foi desenvolvido com o intuito de auxiliar na consolidação óssea. Estudos anteriores sugerem que a combinação de substitutos ósseos e células-tronco mesenquimais (CTM) podem auxiliar a potencializar e promover a regeneração óssea. No presente estudo, o biomaterial foi desenvolvido e a viabilidade e a citotoxicidade de células Vero (ATCC CCL-81) e CTM obtidas de medula óssea provenientes de ovinos utilizando ensaios metil-tiazol-tetrazólio, MTT e iodeto de propídeo (PI) foram avaliadas em diferentes concentrações de extrato desse compósito. O compósito demonstrou ausência de citotoxicidade com comportamento semelhante ao grupo controle. Amostras com 50% e 10% de concentração de extrato do compósito mostraram resultados maiores comparados ao grupo controle (ensaio MTT). Esses resultados também sugerem que a presença do biomaterial pode ser utilizada em associação a CTM. Assim, esse estudo conclui que o compósito apresentado de hidroxiapatita, quitosana e nanotubo de cabono não foi considerado citotóxico e pode ser utilizado em teste in vivo.(AU)

Chitosan and gelatin materials incorporated with phenolic extracts of grape seed and jabuticaba peel: Rheological, physicochemical, antioxidant, antimicrobial and barrier properties.

In this study, chitosan and gelatin materials incorporated with grape seed (Vitis vinifera L.) (VSE) and jabuticaba peel (Plinia cauliflora) (PPE) extracts were developed with potential application as food coatings. It was evaluated how the concentration of the extracts and their addition order in the polymeric matrix affect its properties. Samples with VSE presented a higher total phenolic content and also a more elastic behavior than samples with PPE. The addition order effect over viscosity was the opposite for the extracts, and for the samples with VSE a lower viscosity was obtained when the extract was added before gelatin. All samples were hydrophilic, a good result for application as coatings. Films with PPE were less soluble than chitosan/gelatin film, and CG5P sample was chosen as the most suitable for the desired application, due to its lower water vapor permeation value. The microbial permeation test showed that all samples avoid microorganism growth, extending shelf life of food. The results of this study revealed the extracts concentration was the main factor which influenced the studied parameters; however, their addition order had significant importance on rheological and barrier properties, the ones most influenced by the availability of extract compounds in the polymeric system.

Elastin-derived scaffolding associated or not with bone morphogenetic protein (BMP) or hydroxyapatite (HA) in the repair process of metaphyseal bone defects.

Tissue engineering represents a promising alternative for reconstructive surgical procedures especially for the repair of bone defects that do not regenerate spontaneously. The present study aimed to evaluate the effects of the elastin matrix (E24/50 and E96/37) incorporated with hydroxyapatite (HA) or morphogenetic protein (BMP) on the bone repair process in the distal metaphysis of rat femur. The groups were: control group (CG), hydrolyzed elastin matrix at 50°C/24h (E24/50), E24/50 + HA (E24/50/HA), E24/50 + BMP (E24/50/BMP), hydrolyzed elastin matrix at 37°C/96h (E96/37), E96/37 + HA (E96/37/HA), E96/37 + BMP (E96/37/BMP). Macroscopic and radiographic analyses showed longitudinal integrity of the femur in all groups without fractures or bone deformities. Microtomographically, all groups demonstrated partial closure by mineralized tissue except for the E96/37/HA group with hyperdense thin bridge formation interconnecting the edges of the ruptured cortical. Histologically, there was no complete cortical recovery in any group, but partial closure with trabecular bone. In defects filled with biomaterials, no chronic inflammatory response or foreign body type was observed. The mean volume of new bone formed was statistically significant higher in the E96/37/HA and E24/50 groups (71.28 ± 4.26 and 66.40 ± 3.69, respectively) than all the others. In the confocal analysis, it was observed that all groups presented new bone markings formed during the experimental period, being less evident in the CG group. Von Kossa staining revealed intense calcium deposits distributed in all groups. Qualitative analysis of collagen fibers under polarized light showed a predominance of red-orange birefringence in the newly regenerated bone with no difference between groups. It was concluded that the E24/50 and E96/37/HA groups promoted, with greater speed, the bone repair process in the distal metaphysis of rat femur.

Use of an anionic collagen matrix made from bovine intestinal serosa for in vivo repair of cranial defects.

Polymeric biomaterials composed of extracellular matrix components possess osteoconductive capacity that is essential for bone healing. The presence of collagen and the ability to undergo physicochemical modifications render these materials a suitable alternative in bone regenerative therapies. The objective of this study was to evaluate the osteogenic capacity of collagen-based matrices (native and anionic after alkaline hydrolysis) made from bovine intestinal serosa (MBIS). Twenty-five animals underwent surgery to create a cranial defect to be filled with native and anionic collagen matrixes, mmineralized and non mineralized. The animals were killed painlessly 6 weeks after surgery and samples of the wound area were submitted to routine histology and morphometric analysis. In the surgical area there was new bone formation projecting from the margins to the center of the defect. More marked bone neoformation occurred in the anionic matrices groups in such a way that permitted union of the opposite margins of the bone defect. The newly formed bone matrix exhibited good optical density of type I collagen fibers. Immunoexpression of osteocalcin by osteocytes was observed in the newly formed bone. Morphometric analysis showed a greater bone volume in the groups receiving the anionic matrices compared to the native membranes. Mineralization of the biomaterial did not increase its osteoregenerative capacity. In conclusion, the anionic matrix exhibits osteoregenerative capacity and is suitable for bone reconstruction therapies.

Influence of collagen addition on the thermal and morphological properties of chitosan/xanthan hydrogels.

This study investigates the collagen influence on thermal and morphological characteristics of chitosan/xanthan hydrogels for potential tissue engineering applications. Anionic collagen was prepared by selective hydrolysis of type I collagen found in bovine tendons. Chitosan was obtained from the partial deacetylation of squid pen ß-chitin and xanthan was acquired from Fluka. The hydrogels were obtained in different ratios and were characterized by thermal and morphological analysis. FT-IR suggested only electrostatic interactions between NH3(+) groups of chitosan and COO(-) groups of xanthan and collagen. Thermogravimetric curves showed that hydrogels contain a great amount of water (above 98%) and the presence of collagen does not change this characteristic. Freezing-bound water transition in DSC curves was shifted to higher values due to the increase of water/polymer interaction, mainly when different ratios of chitosan and xanthan were used. SEM images showed sheet-form structures with the presence of collagen promoting an increase in pore size.

Implantação de biomembrana de colágeno tratada em solução alcalina ou conservada em glicerina a 98% na parede abdominal de equinos / Implantation of collagen biomembrane treated in alkaline solution or conserved in 98% glycerin in the abdominal wall of horses

Objetivou-se, neste trabalho, obter biomembranas de colágeno tratadas em solução alcalina por 72 horas (GE), a partir de centros tendinosos diafragmáticos de equinos e comparar sua biocompatibilidade com membranas conservadas em solução de glicerina a 98% (GG) e membranas não tratadas (GC). As membranas foram implantadas na fáscia interna do músculo reto do abdome de equinos e retiradas, juntamente com os tecidos adjacentes, aos sete, 63 e 126 dias pós-operatórios para a preparação de lâminas histológicas. O estudo histomorfométrico das lâminas revelou processo inflamatório mais intenso para os implantes GG e CC e cicatrização mais rápida para os implantes GE. Concluiu-se que as biomembranas colagênicas tratadas em solução alcalina são mais biocompatíveis do que biomembranas conservadas em glicerina 98%.

The objective of this research was obtain collagen biomembranes treated in alkaline solution for 72 hours (GE) from tendineous diaphragmatic center of equines and compare its biocompatibility with membranes preserved in a glycerin solution 98% (GG) and membranes do not treated (GC). The membranes were implanted in the internal fascia of recto abdominis muscle of equines and removed, with adjacent tissues, seven, 63 and 126 days postoperative for the preparation of histological slides. The histomorphometric study revealed more intense inflammatory process to GG and CC implants and faster healing for GE implants. It was concluded that the collagen biomembranes treated in alkaline solution is more biocompatible than biomembranes preserved in 98% glycerin.

Biomateriais para reparação cirúrgica da parede abdominal em animais domésticos: revisão / Biomaterials for surgical repair of abdominal wall in animals: review / Biomateriales para reparación quirúrgica de la pared abdominal en animales domésticos: revisión

A reparação dos tecidos que compõem a parede abdominal é uma necessidade cirúrgica constante. Especificamente para utilização em animais domésticos, o maior desafio é o desenvolvimento de biomateriais que suportem forças exercidas pelas vísceras, em virtude de sua posição quadrupedal e pela movimentação das estruturas após o procedimento cirúrgico. Nas últimas décadas diversos materiais sintéticos têm surgido e apresentado resultados significativos em relação à tensão a que são submetidos e à formação de tecido cicatricial. No entanto, o conhecimento interdisciplinar das estruturas moleculares responsáveis pela antigenicidade, proliferação celular e dos mecanismos que possam alterá-los, proporcionou a obtenção de biomembranas derivadas de tecidos orgânicos, com resultados superiores em relação à interação com os tecidos dos hospedeiros dos implantes e à regeneração tecidual. Assim, o objetivo desta revisão é de conflitar os principais trabalhos de uso comparativo de biomateriais naturais e sintéticos na parede abdominal e compilar os conhecimentos moleculares, que norteiam o desenvolvimento de técnicas de obtenção de biomateriais naturais.

The repairing of abdominal wall tissues is a constant surgical necessity. Specifically for use in domestic animals, the biggest challenge is the development of biomaterials that support forces exerted for the viscera, due to quadruped position and for the movement of the structures after the surgical procedure. In the last decades several synthetic materials have appeared and presented significant results in relation to the tension that are submitted and to the cicatricial tissue formation. However, the interdisciplinary knowledge of antigenic molecular structures, cellular proliferation signs and of the mechanisms that can modify them provided the attainment of biomembranes derived from organic tissues with superior results in relation to the interaction with host tissues and to tissue regeneration. Therefore, the objective of this review is to conflict the main works of comparative use of natural and synthetic biomaterials in the abdominal wall and to compile the molecular knowledge that guide the development of techniques of attainment of natural biomaterials.

La reparación de los tejidos que componen la pared abdominal es una necesidad quirúrgica constante. Específicamente para utilización en animales domésticos, el mayor reto es el desarrollo de biomateriales que soporten fuerzas ejercidas por las vísceras, debido a su posición cuadrupedal y por el movimiento de las estructuras tras el procedimiento quirúrgico. En las últimas décadas, muchos materiales sintéticos han surgido y han mostrado resultados significativos en relación a la tensión a que son sometidos y a la formación de tejido cicatrizal. Sin embargo, el conocimiento interdisciplinario de las estructuras moleculares responsables por la antigenicidad, la proliferación celular y de los mecanismos que pueden alterarlos, resultó a obtener biomembranas derivadas de tejidos orgánicos, con resultados superiores en relación a la interacción con los tejidos huéspedes de los implantes y a la regeneración de los tejidos. Por lo tanto, el objetivo de esta revisión es discutir los principales trabajos de uso comparativo de biomateriales naturales y sintéticos en la pared abdominal y compilar los conocimientos moleculares, que guían el desarrollo de técnicas de obtención de biomateriales naturales.

Membranas biológicas homólogas preservadas em solução alcalina seguida de liofilização, glicerina a 98 por cento e por liofilização para implantação em eqüinos / Homologous biological membranes persevered in alkaline solution and lyophilized, glycerin 98 percent and by lyophilization for equines implantation

Objetivou-se, neste trabalho, avaliar as alterações físico-químicas proporcionadas pelo tratamento de centros tendinosos diafragmáticos homólogos em solução alcalina seguida de liofilização, para implantação na fáscia interna do músculo reto do abdome de eqüinos. As amostras foram tratadas em períodos de 24, 48, 72, 120 e 144 horas, liofilizadas e analisadas quanto à homogeneidade, flexibilidade e resistência à sutura das amostras. Posteriormente foram caracterizadas por calorimetria exploratória diferencial e microscopia eletrônica de varredura. Para a implantação nos eqüinos, foram utilizadas amostras tratadas por 72 horas seguidas de liofilização, amostras conservadas em glicerina 98 por cento e amostras apenas liofilizadas, que foram retiradas após uma, nove e 18 semanas para avaliar a existência de aderências. Verificou-se que a homogeneidade e a flexibilidade são diretamente proporcionais ao aumento do tempo de tratamento em solução alcalina, enquanto que a resistência é inversamente proporcional ao aumento de tempo, sendo o tratamento por 72 horas intermediário para estas características. A calorimetria exploratória diferencial mostrou que o tratamento não desnatura o colágeno presente nas amostras. Na microscopia eletrônica de varredura, observou-se que o aumento de tempo de tratamento proporciona expansão de zonas menos densas do material. Em relação à formação de aderências, as amostras apenas liofilizadas apresentaram grau máximo na formação da classificação proposta, seguida pelas amostras conservadas em glicerina 98 por cento com grau médio e as amostras tratadas em solução alcalina e liofilizadas, que foram classificadas em grau mínimo. Concluiu-se que o tratamento por 72 horas seria mais apropriado para implantação e que a integração tissular com a parede abdominal foi melhor em relação às amostras apenas liofilizadas e às conservadas em glicerina.

The objectives in this research were to evaluate physicist-chemistries alterations in equines diaphragmatic tendineous centers implants submitted to alkaline solution and lyophilization treatment. The samples had been treated in alkaline solution for 24, 48, 72, 120 and 144 hours and lyophilized. It was analyzed the homogeneity, flexibility and resistance to the suture. After that, they had been submitted to the differential scanner calorimetry and to the scanner electronic microscopy. Samples of 72 hours in alkaline treatment, conserved in glycerin 98 percent and only lyophilized were implanted in recto abdominis internal fascia of equines. They were removed one, nine and 18 weeks post-implantation to evaluate the existence of tacks. It was verified that the homogeneity and flexibility are directly proportional to the increase of time of treatment in alkaline solution while the resistance is inversely proportional to the time increase, being 72 hour treatment intermediate for these characteristics. The differential scanner calorimetry showed that the treatment do not denature the present collagen in samples. The electronic microscopy showed that the increase of treatment time provides expansion of less dense zones of the material. In relation to the formation of tacks, the samples only lyophilized had presented maximum degree in the proposal classification followed for the conserved in glycerin 98 percent samples with average degree and the treated in alkaline solution and lyophilized samples had been classified in minimum degree. It was concluded that the treatment for 72 hours would be more appropriate for implantation and that the tissue integration with abdominal wall was better in relation to the samples lyophilized only and to the conserved in glycerin ones.

Avaliação do grau de conversão monomérica de resina composta fotopolimerizada por leds / Degree of conversion evaluation of a resin composite polymerized by leds

O objetivo deste estudo foi avaliar o grau de conversão monomérica em diferentes profundidades (1,2,3 mm) e diferentes tempos de armazenagem (0 hora, 168 horas e 336 horas), após cura de uma resina composta (Z-100/3M-ESPE) fotopolimerizada por um aparelho de Leds (Ultrablue IS/DMC) por ativação gradual com intensidade crescente, por 40 segundos (250 a 600 mW/cm²). O grau de conversão foi avaliado por meio de FT-IR, os corpos de prova foram triturados e, posteriormente, prensados com KBr. Os resultados foram avaliados pelo teste estatístico ANOVA e com base neste foi possível observar que não houve diferença estatística significante em relação à profundidade e tempo de armazenagem dos corpos de prova (P<0.05). Conclui-se que a profundidade de polimerização conseguida com o aparelho utilizado não possui diferença estatística significante do primeiro ao terceiro milímetro e para o tempo de armazenagem dos corpos de prova, o grau de conversão não teve diferença estatística significante também, sendo que a conversão monométrica da resina composta ocorreu na presença da ativação física e não se alterou significantemente com o tempo.

Avaliações clínica e morfométrica da capacidade angiogênica da membrana de quitosana em córneas de coelhos / Clinical and morphometric evaluations of the angiogenic capacity of chitosan membrane in rabbit corneas

OBJETIVO: Estudar a vascularização corneal (VC) induzida pela membrana de quitosana (MQ) implantada por enxertia interlamelar na córnea de coelhos. MÉTODOS: Foram utilizados 16 coelhos. No olho esquerdo procedeu-se enxertia interlamelar de um fragmento de 0,25 x 0,25 cm de MQ (olho tratado). No olho direito realizou-se apenas a microbolsa estromal (olho controle). Avaliaram-se clinicamente os animais aos 1, 3, 7, 15 e 30 dias de pós-operatório. Aos 30 dias mensurou-se a VC pelo Sistema Analisador de Imagens LEICA QWIN-550®. RESULTADOS: Aos sete dias observou-se VC a 1,5±0,93 mm do limbo em direção ao eixo visual. Aos 15 dias houve aumento da VC (4,75±3,20 mm), que se manteve aos 30 dias (4,25±4,10 mm). Os olhos controles não apresentaram quaisquer alterações oculares. Houve diferença estatística (p<0,05) entre as áreas corneais vascularizadas dos olhos tratados e controles aos 15 e 30 dias de pós-operatório. CONCLUSÕES: A MQ induziu angiogênese corneal quando aplicada à córnea de coelhos por enxertia interlamelar, a qual persistiu de forma leve até 30 dias de pós-operatório. Embora estudos adicionais sejam necessários a MQ poderá ser mais uma opção de membrana para enxertos em ceratoplastias.

PURPOSE: To evaluate corneal vascularization (CV) induced by interlamellar graft chitosan membrane (CM) in rabbit cornea. METHODS: An interlamellar graft with a 0.25 x 0.25 cm CM fragment was performed in the left eye (treated eye). In the right eye, an estromal tunnel was done (control eye). The clinical evaluation was done at 1, 3, 7, 15 and 30 days postoperatively. CV analysis was after 30 days by the Images Analizator System LEICA QWIN-550®. RESULTS: After 7 days, CV at 1.5±0.92 mm from the limbus in direction of the cornea axial area was observed. After 15 days CV increased (4.75±3.19 mm), remaining until day 30 (4.25±4.06 mm). The control eyes did not present any changes. There was a statistical differences of the vascularizated corneal areas between control and treated eyes from the 15th to the 30th postoperative day. CONCLUSION: The chitosan membrane induced corneal angiogenesis when applied to rabbit cornea through an interlamellar graft, which was maintained at low levels until 30 days postoperatively. Although further studies are necessary, the results found here demonstrated the usefulness of chitosan membrane in keratoplasties.

[Clinical and morphometric evaluations of the angiogenic capacity of chitosan membrane in rabbit corneas]. / Avaliações clínica e morfométrica da capacidade angiogênica da membrana de quitosana em córneas de coelhos.

PURPOSE: To evaluate corneal vascularization (CV) induced by interlamellar graft chitosan membrane (CM) in rabbit cornea. METHODS: An interlamellar graft with a 0.25 x 0.25 cm CM fragment was performed in the left eye (treated eye). In the right eye, an estromal tunnel was done (control eye). The clinical evaluation was done at 1, 3, 7, 15 and 30 days postoperatively. CV analysis was after 30 days by the Image Analyzer System LEICA QWIN-550. RESULTS: After 7 days, CV at 1.5+/-0.92 mm from the limbus in direction of the cornea axial area was observed. After 15 days CV increased (4.75+/-3.19 mm), remaining until day 30 (4.25+/-4.06 mm). The control eyes did not present any changes. There was a statistical differences of the vascularizated corneal areas between control and treated eyes from the 15th to the 30th postoperative day. CONCLUSION: The chitosan membrane induced corneal angiogenesis when applied to rabbit cornea through an interlamellar graft, which was maintained at low levels until 30 days postoperatively. Although further studies are necessary, the results found here demonstrated the usefulness of chitosan membrane in keratoplasties.

Estudos preliminares de liberação de ciprofloxacina em compósito hidroxiapatita: colágeno / Preliminary studies of ciprofloxacin delivery in hydroxyapatite: collagen composite

Compósitos hidroxiapatita: colágeno foram preparados em diferentes proporções a fim de se determinar qual a melhor proporção para a incorporação de antibiótico. A melhor proporção de HA:colágeno obtida foi de 10:1 (m/m), a qual foi caracterizada por Calorimetria Exploratória Diferencial (DSC), espectroscopia no Infravermelho (IV), Microscopia Eletrônica de Varredura (MEV) e teste de hidrofilicidade, sendo então posteriormente utilizada na incorporação e liberação de ciprofloxacina. As curvas calorimétricas mostraram transições em torno de 40 graus C, referentes à desnaturação do colágeno. A razão A1235/A1450 do espectro no infravermelho do compósito HA:col (10:1) foi de 1,12 e mostra que o colágeno está preservado neste compósito. As fotomicrografias mostram fibras de colágeno uniformemente distribuídas em torno da hidroxiapatita. O teste de hidrofilicidade do compósito mostrou que o material foi capaz de absorver 183,2 por cento de água em relação à sua massa seca. Para os estudos preliminares de liberação de antibiótico incorporou-se cerca de 5 por cento de cloridrato de ciprofloxacina em massa no compósito HA:col (10:1), no momento de preparo do mesmo. O experimento de liberação in vitro foi realizado em tampão fostato salino (PBS), pH 7,4 a 37 graus C, durante 72 horas, mostrando uma liberação máxima em torno 90 por cento, após 10 horas de imersão. A liberação da droga nas primeiras quatro horas obedece ao modelo de Higuchi, sendo indicativo de processo de difusão pelos poros da matriz