Canine umbilical cord perivascular tissue: A source of stem cells for therapy and research.

There are numerous sources of multipotent mesenchymal stromal cells (MSC) with therapeutic potential, and bone marrow is the main one. However, pain, lack of donors and comorbidities associated with harvesting stimulate the search for new sources of MSCs. The aim of this work is to obtain cells from umbilical cord (UC) perivascular tissue of dogs and characterize them as MSCs. For this, the UC was obtained from therapeutic cesarean sections and submitted to enzymatic digestion. The obtained cells were subjected to growth and proliferation tests, as well as the analysis of surface markers, differentiation test in three mesenchymal lineages and analysis of differentiation markers expression. From all the UC used in this study an adherent with fibroblastoid shape cell was obtained, with an initial number of 4.8 × 105 of cells. The growth curves showed a lag phase from 0 to 24 h, followed by a phase of growth of 24 to 168 h, and then phase of cell decay. The doubling time was kept around 15 h until the sixth passage, from which there were signs of cellular senescence. The differentiation assays demonstrated the ability of cells to differentiate into osteoblasts, adipocytes and chondrocytes when subjected to the induction mediums. The study of surface markers was positive for adhesion markers and negative for hematopoietic markers. Thus, cells obtained from canine UC perivascular tissue by enzymatic digestion are multipotent MSC and the protocol developed ensures the perivascular origin of these cells.

Hydroxyapatite-lignin composite as a metallic implant-bone tissue osseointegration improver: experimental study in dogs / Compósito de hidroxiapatita sintética e lignina como promotor de osseointegração entre implante metálico e tecido ósseo: estudo experimental em cães

The study aimed to evaluate biocompatibility, osteoconduction and osseointegration of a pasty composite of hydroxyapatite (20%) and lignin (80%) as a promoter of metal implant and bone tissue integration. An intramedullary Schanz pin was implanted in both tibias of fifteen bitches. In the left tibia, the pin was coated with the biomaterial at the time of surgery. Marrow cavity was also filled with the biomaterial. Right limb did not receive the biomaterial, then constituting the control group. Tibias were harvested from five animals at 8, 60 and 150 days after surgery; three of them were analyzed by histological and biomechanical assessment and the two remaining tibias by X-ray diffraction. Results showed that the biomaterial is biocompatible, with osteoconductivity and osseointegration properties. Histological analysis and diffractograms showed the presence of hydroxyapatite in samples in all periods, although the presence of organic material of low crystallinity was variable. There was no statistical difference in the forces required for removal of the biocompatibility, osteoconductivity and osseointegration, it was not able to promote a better intramedullary pin anchorage.

O trabalho objetivou avaliar a biocompatibilidade, a osteocondução e a osseointegração de um compósito pastoso de hidroxiapatita (20%) e lignina (80%) como promotor de integração entre implante metálico e tecido ósseo. Um pino intramedular de Schanz foi implantado em ambas as tíbias de quinze cadelas. Na tíbia esquerda, o pino foi coberto com o biomaterial no momento da cirurgia. A cavidade medular também foi preenchida com o biomaterial. O membro direito não recebeu o biomaterial, constituindo assim o grupo controle. As tíbias foram coletadas de cinco animais aos oito, 60 e 150 dias após a cirurgia, sendo três encaminhadas para avaliação histológica e biomecânica e as duas restantes, para difração de raios X. Os resultados revelaram biocompatibilidade do material, com propriedades de osteocondução e osseointegração. As análises histológicas e os difratogramas mostraram presença da hidroxiapatita no tecido em todos os períodos avaliados, embora a presença de material orgânico de baixa cristalinidade tenha sido variável. Não houve diferença estatística na força necessária para remoção do pino intramedular entre os tempos ou tratamentos. Embora o biomaterial tenha apresentado biocompatibilidade, osteocondutividade e osseointegração, ele não foi capaz de promover uma melhor ancoragem do pino intramedular.

Guided tissue regeneration using rigid absorbable membranes in the dog model of chronic furcation defect.

OBJECTIVE: Absorbable membranes are used to promote the regeneration of periodontal defects by Guided Tissue Regeneration (GTR). However, their collapse into the defect is commonly reported, impairing regeneration. Therefore, absorbable but rigid membranes aiming at preventing such collapse were developed and analyzed in periodontal regeneration. MATERIALS AND METHODS: Membranes were analyzed in class II furcation defects in dogs; procedures included periodontal disease induction, prophylaxis and GTR (treated groups) or open flap debridement alone (control group). For GTR, the membranes were made of either 25% hydroxyapatite (HA) in polyhydroxybutyrate matrix (PHB) or 35% HA in PHB. Animals were clinically evaluated for gingival recession, clinical attachment level (CAL) and biopsies were collected at 60 and 120 days. Bone volume, trabeculae number, trabecular thickness and trabecular separation were quantified by micro-computed tomography, followed by histology. RESULTS: Membrane exposure was observed in both treated groups (25 and 35% HAP) from the 8(th) day after surgery, continuously progressing until 120 days. Mean CAL for all groups remained above normal values for dogs. Bone volumetric values were not significantly different. Partial formation of bone, cementum and periodontal ligament was observed in treated groups. An inflammatory infiltrate was observed in the dense connective tissue that partially filled the center of the treated defects with active osteoclasts on bone surface. CONCLUSION: Although partial regeneration of the defect was observed, it was limited by wound contamination. Consequently, rigid absorbable membranes made of HA and PHB failed to improve the regeneration of class II furcation defects in dogs.