Surface topography of resorbable porcine collagen membranes, and their effect on early osteogenesis: An in vitro study.

OBJECTIVE: Guided tissue regeneration (GTR) is based on the use of different membranes that function as sealants and barriers in specific clinical situations. Among the several tissue production methods and origins, resorbable porcine-derived membranes are the most commonly used. Because these membranes are so diverse, and have several different clinical applications, doubts linger as to their effect in stimulating osteogenesis. The objective of this study was to make an in vitro evaluation of the viability and differentiation of osteoblastic cells cultured on the surface of the following collagen membranes: Jason® (Botiss Biomaterials), Collprotect® (Botiss Biomaterials), and Bio-Gide® (Geistlich). MATERIAL AND METHODS: Fragments of the 3 resorbable collagen membranes (5 × 5 mm) were used, and pre-osteoblastic SAOS-2 cells (ATCC, USA) were plated on their porous surfaces. Evaluation of the membranes was performed at 3, 5 and 7 days, considering the following parameters: (1) topographic analysis of the different surfaces by scanning electron microscope; (2) cellular viability by MTT, (3) quantification of type I collagen and osteopontin by Elisa. The quantitative analyses were carried out using a significance level of 5%. RESULTS: Collprotect® and Jason® membranes presented a rough surface with an irregular aspect on both sides, while double-layered Bio-Gide® had one layer with a smooth surface and the other with a rough surface along each respective length. The viability assays revealed that the cells cultured the cells grown on Collprotect® showed higher viability than those grown in Bio-Gide® or Jason®, especially after 5 and 7 days. After 3 and 5 days, evaluation of type I collagen showed that the cells plated on the Jason® and Collprotect® surfaces had greater collagen secretion than those plated on BioGide®. After 7 days, an increase in osteopontin levels was observed when the cells were plated on all the experimental membranes, compared with the control group. CONCLUSION: All the tested membranes were suitable for use in GTR clinical procedures. Their indication in specific regenerative cases depends on the mechanical and biological properties of their originating tissues, thus enabling better results and assertive choices by dental professionals.

Comparison of Two Bovine Commercial Xenografts in the Regeneration of Critical Cranial Defects.

Autologous bone is the gold standard in regeneration processes. However, there is an endless search for alternative materials in bone regeneration. Xenografts can act as bone substitutes given the difficulty of obtaining bone tissue from patients and before the limitations in the availability of homologous tissue donors. Bone neoformation was studied in critical-size defects created in the parietal bone of 40 adult male Wistar rats, implanted with xenografts composed of particulate bovine hydroxyapatite (HA) and with blocks of bovine hydroxyapatite (HA) and Collagen, which introduces crystallinity to the materials. The Fourier-transform infrared spectroscopy (FTIR) analysis demonstrated the carbonate and phosphate groups of the hydroxyapatite and the amide groups of the collagen structure, while the thermal transitions for HA and HA/collagen composites established mainly dehydration endothermal processes, which increased (from 79 °C to 83 °C) for F2 due to the collagen presence. The xenograft's X-ray powder diffraction (XRD) analysis also revealed the bovine HA crystalline structure, with a prominent peak centered at 32°. We observed macroporosity and mesoporosity in the xenografts from the morphology studies with heterogeneous distribution. The two xenografts induced neoformation in defects of critical size. Histological, histochemical, and scanning electron microscopy (SEM) analyses were performed 30, 60, and 90 days after implantation. The empty defects showed signs of neoformation lower than 30% in the three periods, while the defects implanted with the material showed partial regeneration. InterOss Collagen material temporarily induced osteon formation during the healing process. The results presented here are promising for bone regeneration, demonstrating a beneficial impact in the biomedical field.

Ordered collagen membranes: production and characterization.

A collagen membrane with microscopic order is presented. The membranes were produced with acid-soluble collagen, using two different methods to obtain orientation. The product was characterized by mean of UV and IR spectra, scanning electronic microscopy, optical microscopy and laser diffractometry. The results clearly show a high level of order in the membranes obtained by both techniques. Permeability for rifamycin, ascorbic acid and NaCl was also measured. Due to the characteristics of the membranes, they have a potential application for treatment of surface injuries.

Cytotoxicity of bovine and porcine collagen membranes in mononuclear cells

This study compared the cytotoxicity and the release of nitric oxide induced by collagen membranes in human mononuclear cells. Peripheral blood was collected from each patient and the separation of mononuclear cells was performed by Ficoll. Then, 2x10(5) cells were plated in 48-well culture plates under the membranes in triplicate. The polystyrene surface was used as negative control. Cell viability was assessed by measuring mitochondrial activity (MTT) at 4, 12 and 24 h, with dosage levels of nitrite by the Griess method for the same periods. Data had non-normal distribution and were analyzed by the Kruskal-Wallis test (p<0.05). Statistically significant differences (p<0.05) were observed between the membranes and the control in the experimental period, although there was a significant reduction in viability over time (p<0.01). At 4 and 12 h, the porcine membrane induced a higher release of nitrite compared with the control and bovine membrane, respectively (p<0.01), and this difference was maintained at 24 h (p<0.05). This in vitro study showed that the porcine collagen membrane induces an increased production of proinflammatory mediators by mononuclear cells in the first hours of contact, decreasing with time.

Neste estudo foi comparada a citoxicidade e a liberação de nitrito induzidos por membranas de colágeno bovino e suíno em células mononucleares humanas. Foram coletados sangue periférico de cada paciente, e realizada separação de mononucleares por gradiente de Ficoll. Um total de 2x10(5) células foram plaqueadas em placas de cultura de 48 poços sob as membranas, em triplicata. O poço sem membrana serviu como controle negativo. A viabilidade celular foi avaliada medindo a atividade mitocondrial (MTT) em 4,12 e 24 h, com dosagens dos níveis de nitrito pelo método de Griess nos mesmos períodos. As amostras não apresentaram distribuição normal, sendo realizado o teste de Kruskal-Wallis (p<0,05). Foram observadas diferenças estatisticamente significantes entre as membranas e o controle nos período analisados (p<0,05), embora tenha ocorrido redução da viabilidade em função do tempo (p<0,01). Em 4 e 12 h a membrana suína induziu maior liberação de nitrito comparado ao controle e à membrana bovina, respectivamente (p<0,01). Tal diferença foi mantida em 24 h (p<0,05). Este estudo in vitro demonstrou que a membrana colágena suína induz uma maior produção de mediador pró-inflamatório pelas células mononucleares nas primeiras horas de contato, diminuindo com o tempo.