Abstract Bone morphogenetic proteins (BMPs) constitute a large family of osteoinductive proteins. Different BMPs are widely used in animal experiments and increasingly in the field of bone surgery. However, the sterilization of BMPs and the choice of a suitable mode of delivery, which binds and slowly releases BMP molecules, are still under intensive investigation. The aims of this study were to evaluate the effects of ethylene oxide and gamma sterilizations and different delivery systems on the osteoinductivity of reindeer BMP extract by using heterotopic and orthopic animal models. Ethylene oxide gas (Steri-Vac 4XL, temperature 29 °C, exposure time 4 h, concentration 860 mg/l) and gamma (doses of 3.15 or 4.15 Mrad) sterilizations were used. The tested delivery systems for reindeer BMP were collagen (Lyostypt®), gelatine capsule (no.1) and composites containing collagen combined with tricalcium phosphate (TCP) or hydroxyapatite (HA) or biphasic tricalcium phosphate-hydroxyapatite (TCP/HA) or biocoral (NC) frames. The injectability of reindeer BMP was tested by using injections containing a saline or gelatine vehicle. Osteoinductivity was evaluated as ectopic bone formation in the thigh muscle pouches of mouse hind legs. The induced new bone was evaluated based on the incorporation of 45Ca or calcium yield, radiographs and histological examination three weeks after the operations. The effect of gamma sterilization on the bone healing capacity of reindeer BMP extract was evaluated in a rabbit radial bone defect model in comparison with non-sterilized reindeer BMP extract and recombinant BMP-2. Bone healing was evaluated after eight weeks based on radiographs, mechanical tests and peripheral computerized tomography (pQCT). All BMP implants induced new bone in vivo visible in radiographs, but no bone formation was seen in the control implants without reindeer BMP. Gamma sterilization did not decrease significantly the osteoinductivity of reindeer BMP extract, except when administered as an injection containing gelatine vehicle. Ethylene oxide sterilization decreased significantly the osteoinductivity of reindeer BMP extract and was significantly inferior compared to gamma sterilization. Reindeer BMP combined with collagen or composite containing collagen and TCP/HA frame induced new bone significantly better than reindeer BMP combined with composite containing collagen and TCP frame. BMP injections with gelatine or saline vehicles induced new bone effectively. Injections were easy to handle and well tolerated by the mice. Reindeer BMP extract administered with collagen carrier healed the bone defect of the rabbit radius significantly better than control implants without reindeer BMP or no treatment and its effect was comparable with rhBMP-2 treatment.

Consensus in scientific literature is that hyperglycemia, which is a condition that manifests in individuals with uncontrolled diabetes, causes compromised bone growth, but the exact mechanisms of are unknown. It has been estimated that 5% of dental implant failures that have previously been linked to unknown causes may be associated with undiagnosed diabetes. It is important to study the early stages of bone growth as it is accepted that they are critical in the long-term success rate of endosseous implants. This study aimed to investigate the bone healing seen in the hyperglycemic group compared to the normal (i.e. control) group, at an early time point, using high-resolution microscopy techniques. Ten young (200-250gram) male Wistar rats were used for this study with five rats assigned to the control group and the other five rats intravenously injected with 65 mg/kg of streptozotocin (STZ) to induce diabetes. An osteotomy model was used to make a 1.3mm defect in the diaphysis of the rat femurs. After five days, the femurs were removed, fixed in glutaraldehyde, dehydrated, and embedded in resin. Structural and chemical analyses were conducted on the samples using a variety of microscopy techniques to examine various factors of bone quality including: bone porosity, relative mineralization level, and the arrangement of collagen and mineral. When analyzing the micro-structure, the hyperglycemic group showed increased porosity in the newly formed bone as compared to the control group. However, no significant differences were found in the nano-structure when analyzing the arrangement of collagen and mineral.Therefore, the results in this thesis suggest that alterations in micro-architecture rather than nano-architecture may play a pivotal role in the compromised bone healing in uncontrolled diabetes at this five-day time point. Future work should investigate additional time points in the bone healing process. / Thesis / Master of Applied Science (MASc) / According to the International Diabetes Federation, 387 million people worldwide are living with diabetes of which 46.3% are undiagnosed. Uncontrolled diabetes results in hyperglycemia, which is a condition where there is an increased level of glucose in the blood. When diabetes is not regulated correctly with medication, it leads to problems in the long-term success rate of dental implants. The objective of this thesis was to investigate the early stages of bone formation, which are accepted to be critical in the long-term success rate of dental implants, in hyperglycemic animal models compared to control groups using various microscopy techniques. The different techniques used allowed for the structural and elemental compositions of bone to be studied on the micro-scale and nano-scale. It was shown that at the 5-day healing time point studied, the micro-structure, rather than the nano-structure, was negatively altered in the hyperglycemic group compared to the control group.

Knochen ist ein multifunktionales Organ und zugleich ein biologisches Material. In dieser Arbeit wird der Heilungsverlauf eines Knochenbruchs (als biologisches Material) näher untersucht mit Hilfe von Computermodellen. Im menschlichen Körper kommt es nach einem Bruch zu einer vollständigen Regeneration des Knochens, ohne dass eine Narbe nach der Heilung zurückbleibt. In grob 10% der Frakturen kommt es jedoch zu Komplikationen bis zu einem Nicht-Heilen des Bruches. Das Ziel von intensiver interdisziplinärer Forschung ist es daher, nicht nur die medikamentöse Behandlung solcher Komplikationen zu verbessern, sondern auch durch externe, biophysikalische Stimulation die Heilung anzuregen. Gewöhnlich heilt ein Knochenbruch nicht direkt (Primäre Knochenheilung), das heißt durch Bildung von neuem Knochen im Knochenspalt, sondern über Sekundäre Knochenheilung. Während der sekundären Heilung bildet sich vorübergehend zusätzliches Gewebe außerhalb des Frakturspaltes, der so genannte Kallus, der die Aufgabe hat, den Bruch zu stabilisieren. Im Kallus werden im Laufe der Heilung verschiedene Gewebearten gebildet (z.B. Bindegewebe, Knorpel und Knochen). Die Gewebe werden von spezialisierten biologischen Zellen gebildet. Die spezialisierten Zellen entwickeln sich aus mesenchymalen Stammzellen (d.h. sie differenzieren), die in den Kallus wandern. Hauptziel der Arbeit ist das bessere Verständnis der mechano-biologischen Regulation der Gewebeformation während der Heilung eines normalen Knochenbruches. Dazu wurden Computersimulationen durchgeführt und mit experimentellen Daten eines Schafmodels verglichen. / Bone is a multifunctional organ, a biological material and is able to fully restore bone fractures without leaving a scar. However, in about 10% of the bone fractures, healing does not lead to a successful reunion of the broken bone ends. Intensive interdisciplinary research therefore looks for new ways to promote healing not only by medication, but also by external biophysical stimulation. Usually, bone fractures do not heal by a direct bridging of the fracture gap with newly formed bone (primary bone healing). Instead, secondary bone healing proceeds indirectly via the formation of an external callus (additional tissue). Within the callus, intricate tissue type patterns are formed, which evolve during the healing progression. Stem cells differentiate into specialized cells, which lay down different tissues such as fibrous tissue, cartilage and bone. This cell differentiation can be biophysically stimulated, e.g. by mechanical deformation of the cytoskeleton. The main aim of this thesis was to connect the microscopic cell response to mechanical stimulation with the macroscopic healing progression. Simple rules for cell behaviour were implemented in a computer model, the progression of healing was simulated and the outcome of the simulations was compared to results from animal experiments. In comparison to existing simulations of bone healing, this study approached the problem from a more physical viewpoint and linked experimental in vivo data and computer modelling.

Fracture healing requires a series of events including inflammatory response and callus formation, callus remodeling and bone healing. Fracture healing is a complex process, there are several overlapping phases , including inflammation , cartilage formation and bone remodeling, there are many internal or external factors could impact on fracture healing, leading to delayed bone healing or non healing. The global androgen receptor knockout (GARKO) mice has been know to reduce bone mass in endochondral bone and osteoblast mineralization, but the impact for callus formation in fracture healing is still unclear. The goals of study is to investigate the role of androgen and androgen receptor in wild-type (WT) mice and GARKO mice after fracture healing during callus formation and bone mineralization and bone remodeling. Therefore, long-term animal experiments observed by micro-computed tomography to study the roles of androgen and androgen receptor on the process and mechanisms of fracture healing is necessary. We applied in vivo micro-computer tomography (Micro-CT) to build up the three-dimensional model images at different time points for wild-type mice and GARKO mice after fracture healing and observe the bone healing process of micro-structure of the development of callus during fracture healing. The callus tissue morphology observed by histological staining to study the proportion and position of collagen, fibrous tissue and bone. The results show that the healing of WT mice is better than GARKO mice. GARKO mice develop smaller callus size and less bone volume and show delayed healing. In general, orchiectomy (ORX) decreases callus size in WT mice but not in GARKO mice. However, the healing rate of elderly GARKO mice is not obvious in comparison with young GARKO mice. Together, our study demonstrated that the androgen and androgen receptor regulate fracture healing and play an important role in bone repair and healing. Our mouse model may be used for the therapeutic drug screening of bone fractures caused by osteoporosis.

A utilização de enxertos ósseos vem aumentando bastante em pequenos animais, entretanto, em aves esta utilização é menor devido à dificuldade de conseguir autoenxerto em quantidades adequadas. Sendo assim, o objetivo do projeto foi avaliar três tipos de enxertos em aves. Foram utilizadas 30 galinhas separadas em três grupos: grupo aloenxerto congelado em ultrafreezer (GUF) por duas semanas; grupo aloenxerto congelado em nitrogênio líquido (GNL) e grupo enxerto sintético de hidroxiapatita deficiente em cálcio (GHA). Os enxertos foram aplicados em defeito ósseo de aproximadamente 20mm na ulna das aves e fixados ao leito receptor com uma placa e quatro parafusos bloqueados. Os animais foram acompanhados por meio de avaliação clínica, radiográfica, histológica e biomecânica. A consolidação radiográfica ocorreu em 90% nos animais do GNL, 70% nos do GUF e 60% nos do GHA, com média de 47,14±13,50 dias no GUF, 61,67±21,79 dias no GNL e aos 70±18,17 dias no GHA, havendo diferença significativa entre o GUF e GHA. Histologicamente os enxertos do GUF apresentavam-se em processo de consolidação mais avançada, com remodelação óssea e a presença de osteoclastos. Na avaliação clínica todas as ulnas operadas tinham estabilidade, sem sinal de inflamação/contaminação, articulações preservadas, sem dor e sem desvios angulares graves na asa. Na análise biomecânica houve diferença significativa (p<0,05) entre o GNL e o GHA na resistência, já na avaliação de flexão máxima, observou-se diferença estatística entre o GHA e o GNL e entre o GHA e o GUF. Sendo assim, é possível concluir que os três enxertos testados podem ser utilizados em defeitos ósseos em aves, mas o melhor deles foi o GNL por apresentar mais vantagens na preparação, confecção, aplicação dos enxertos e resistência biomecânica, com maior taxa de consolidação óssea. / The use of bone grafts is increasing considerably in small animals, however, in poultry this use is less due to the difficulty of obtaining autograft in adequate quantities. Therefore, the objective of the project was to test three types of grafts, evaluating their best results over a period of 90 days. For this purpose, 30 chickens were divided into three groups: frozen ultra freezer allograft group (UFG) for two weeks; allograft frozen in liquid nitrogen group (LNG) and synthetic grafts of calcium deficient hydroxyapatite group (HAG). Grafts were applied to a bone defect of approximately 20 mm in the ulna of birds, in all three groups the grafts were fixed to the recipient bed with plates blocked and 4 screws locked. The animals were followed up by serial radiographs, histological examination, clinical orthopedic evaluation and biomechanical flexion assay. Radiographic consolidation occurred in 90% of the LNG, 70% in the GUF and 60% in the GHA, mean of 47.14 ± 13.50 days without UFG, 61.67 ± 21.79 days non LNG and at 70 ± 18.17 days without HAG, with a significant difference between UFG and HAG. Histologically, UFG grafts were in a more advanced consolidation process, presenting bone remodeling with a presence of osteoclasts In the clinical review, all of them were operated with stability, with no sign of inflammation / contamination, preserved joints, no pain and no angular deviations, graves in the nail. In the biomechanics, there was a significant difference (p <0.05) between LNG and HAG in the resistance, already in the evaluation of maximum flexion, it was observed statistical difference not HAG with LNG and HAG with UFG grafts. Thus, it is possible to conclude that the three grafts tested are feasible to be used in bone loss in birds, where the LNG was superior because it presented more advantages in the preparation, preparation and application of the grafts, presenting a higher rate of bone consolidation.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The main reason demanding the veterinary orthopedics services are fractures. There are several alternatives for the treatment of a fracture; however all of them aim to obtain a fast bone union, preserving the normal function of joints and soft tissues close to it. Some fractures consolidate adequately, after the correct use of a specific immobilization method, yet others result in a slower consolidation or no union. The repair of a fracture involves a sequence of cell events which evolutes since the aggression itself, formation of hematoma, starting of a plastic callus, organization of callus and recasting. It is proposed in this research, a technique for external immobilization of radio and ulna fractures in dogs. This method was used in 16 animals, patients from The Veterinary Hospital from Federal University of Santa Maria (HVU-UFSM) during the period from March 2008 to May 2009. All the patients were clinically evaluated concerning their general health conditions; after this evaluation, it was obtained the clinical-surgical diagnosis of the fracture with the help of a radiographic exam of the fractured limb. After being submitted to general anesthesia, the closed reduction of the fracture was performed, which was immobilized using an aluminum cast, made and adapted exactly to the anatomical shape of the animal s limb. Aiming to accompany the process of bone healing, the patients were submitted to radiographic control at 30, 45 and 60 days after treatment and the cast was removed when a stable bone callus was seen and there was not the line of fracture anymore. The method proposed provided the necessary conditions for the healing process, allowing, thus, the bone consolidation and the return to the patient s normal ambulation and can be used as an additional alternative to the external immobilization methods of radio and ulna fractures in dogs. / A principal causa de procura pelo setor de ortopedia veterinária são as fraturas. Existem várias alternativas para o tratamento de uma fratura, porém todas têm por finalidade obter rápida união óssea, preservando a função normal das articulações e dos tecidos moles próximos. Algumas fraturas consolidam adequadamente após uso correto de determinado método de imobilização, contudo outras resultam em consolidação demorada ou não-união. A reparação de uma fratura envolve uma sequência de eventos celulares que evoluem desde agressão propriamente dita, formação do hematoma, iniciação do calo plástico, organização do calo e remodelação. Propõe-se nesta pesquisa uma técnica alternativa para imobilização externa de fraturas de rádio e ulna em cães. Esse método foi empregado em 16 animais pacientes do Hospital Veterinário Universitário da Universidade Federal de Santa Maria (HVU UFSM) no período de março de 2008 a maio de 2009. Todos os pacientes foram avaliados clinicamente quanto as suas condições gerais de saúde; posteriormente a esta avaliação, obteve-se o diagnóstico clínico cirúrgico da fratura com auxílio do exame radiográfico do membro fraturado. Após anestesia geral, foi realizada a redução fechada da fratura, a qual foi imobilizada, utilizando-se uma canaleta de alumínio, confeccionada e moldada exatamente aos contornos anatômicos do membro do animal. No intuito de acompanhar o processo de cicatrização óssea, os pacientes foram submetidos ao controle radiográfico aos 30, 45 e 60 dias após o tratamento. Sendo a canaleta removida quando da formação de calo ósseo estável e desaparecimento da linha de fratura. O método proposto proporcionou as condições necessárias para o processo de cicatrização das fraturas, permitindo, assim, a consolidação óssea e o retorno à deambulação normal aos pacientes, podendo ser empregado como alternativa adicional aos métodos de imobilização externa de fraturas de rádio e ulna em cães.

A utilização de enxertos ósseos vem aumentando bastante em pequenos animais, entretanto, em aves esta utilização é menor devido à dificuldade de conseguir autoenxerto em quantidades adequadas. Sendo assim, o objetivo do projeto foi avaliar três tipos de enxertos em aves. Foram utilizadas 30 galinhas separadas em três grupos: grupo aloenxerto congelado em ultrafreezer (GUF) por duas semanas; grupo aloenxerto congelado em nitrogênio líquido (GNL) e grupo enxerto sintético de hidroxiapatita deficiente em cálcio (GHA). Os enxertos foram aplicados em defeito ósseo de aproximadamente 20mm na ulna das aves e fixados ao leito receptor com uma placa e quatro parafusos bloqueados. Os animais foram acompanhados por meio de avaliação clínica, radiográfica, histológica e biomecânica. A consolidação radiográfica ocorreu em 90% nos animais do GNL, 70% nos do GUF e 60% nos do GHA, com média de 47,14±13,50 dias no GUF, 61,67±21,79 dias no GNL e aos 70±18,17 dias no GHA, havendo diferença significativa entre o GUF e GHA. Histologicamente os enxertos do GUF apresentavam-se em processo de consolidação mais avançada, com remodelação óssea e a presença de osteoclastos. Na avaliação clínica todas as ulnas operadas tinham estabilidade, sem sinal de inflamação/contaminação, articulações preservadas, sem dor e sem desvios angulares graves na asa. Na análise biomecânica houve diferença significativa (p<0,05) entre o GNL e o GHA na resistência, já na avaliação de flexão máxima, observou-se diferença estatística entre o GHA e o GNL e entre o GHA e o GUF. Sendo assim, é possível concluir que os três enxertos testados podem ser utilizados em defeitos ósseos em aves, mas o melhor deles foi o GNL por apresentar mais vantagens na preparação, confecção, aplicação dos enxertos e resistência biomecânica, com maior taxa de consolidação óssea. / The use of bone grafts is increasing considerably in small animals, however, in poultry this use is less due to the difficulty of obtaining autograft in adequate quantities. Therefore, the objective of the project was to test three types of grafts, evaluating their best results over a period of 90 days. For this purpose, 30 chickens were divided into three groups: frozen ultra freezer allograft group (UFG) for two weeks; allograft frozen in liquid nitrogen group (LNG) and synthetic grafts of calcium deficient hydroxyapatite group (HAG). Grafts were applied to a bone defect of approximately 20 mm in the ulna of birds, in all three groups the grafts were fixed to the recipient bed with plates blocked and 4 screws locked. The animals were followed up by serial radiographs, histological examination, clinical orthopedic evaluation and biomechanical flexion assay. Radiographic consolidation occurred in 90% of the LNG, 70% in the GUF and 60% in the GHA, mean of 47.14 ± 13.50 days without UFG, 61.67 ± 21.79 days non LNG and at 70 ± 18.17 days without HAG, with a significant difference between UFG and HAG. Histologically, UFG grafts were in a more advanced consolidation process, presenting bone remodeling with a presence of osteoclasts In the clinical review, all of them were operated with stability, with no sign of inflammation / contamination, preserved joints, no pain and no angular deviations, graves in the nail. In the biomechanics, there was a significant difference (p <0.05) between LNG and HAG in the resistance, already in the evaluation of maximum flexion, it was observed statistical difference not HAG with LNG and HAG with UFG grafts. Thus, it is possible to conclude that the three grafts tested are feasible to be used in bone loss in birds, where the LNG was superior because it presented more advantages in the preparation, preparation and application of the grafts, presenting a higher rate of bone consolidation.

Orientador: Sergio de Toledo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-07-22T13:10:45Z (GMT). No. of bitstreams: 1 Pereira_SergioLuisdaSilva_M.pdf: 4372724 bytes, checksum: d849d535308306a4ec155f540c1f5542 (MD5) Previous issue date: 1997 / Resumo: O objetivo deste trabalho foi avaliar, histológica e radiograficamente, o efeito da calcitonina de salmão sobre a cicatrização de defeitos ósseos circunscritos, criados cirurgicamente em fêmur de coelhos saudáveis. Estes foram divididos aleatoriamente em dois grupos: controle (n=12) e tratado (n=12). O grupo controle não recebeu qualquer substância durante o experimento, enquanto que no grupo tratado foi aplicada dose diária de 2 UI/kg de calcitonina por via intramuscular. Os animais foram sacrificados nos períodos de 7, 14, 21 ou 28 dias após a criação dos defeitos ósseos. As áreas radiolúcidas destes foram medidas, em mm2, através de um programa de computador denominado Auto-Cad, demonstrando que estas, em média, foram menores no grupo tratado, mas estatisticamente significantes apenas nos períodos intermediários de 14 e 21 dias. Histologicamente, aos 7 dias, os defeitos ósseos do grupo tratado apresentaram uma maior neoformação óssea. Aos 28 dias, houve a formação de um osso menos compacto no -grupo controle. No entanto, aos 14 e 21 dias, os resultados foram semelhantes entre os dois grupos. Os resultados obtidos neste trabalho possibilitaram inferir que a calcitonina de salmão acelerou o processo de cicatrização óssea em defeitos circunscritos, baseados nas análises radiográficas e histológicas realizadas / Abstract: The purpose of this study was to evaluate, in healthy animals, the role of salmon calcitonin in the bone healing process of circumscribed bone defects, surgica1ly created, in the twenty-four (24) young adults rabbits femur. Twelve (12) animals were randomized to be the control and test groups (salmon calcitonin - 2 IU/kg during 21 days). The rabbits were sacrificed 7, 14, 21 and 28 days after the surgical procedures. The radiolucids areas of the bone defects was measured, in mm2, by Auto-Cad software. The results showed statistical differences (p<0,05) at 14 and 21 days, favoring the test group. At 7-day, the bone defects of the test group showed more bone regeneration. Nevertheless, at 14 and 21-days, the results was similar among two groups, with the new bone more compact in the test group than control group at 28 days. Thus, this study did demonstrate the positive role of salmon calcitonin in the bone healing process of circumscribed bone defects in healthy rabbits, considering the radioagraphic and histological analysis / Mestrado / Periodontia / Mestre em Clínica Odontológica

Abstract Bone is a highly vascularized tissue which enables a close interaction between blood vessels and osteoid complexes, including osteoblasts and osteoclasts. Iatrogenic bone loss, caused by a surgical intervention or trauma, disrupts circulation in the bone and leads to hypoxia and even necrosis of the adjacent bone. The mechanisms of the bone tissue healing process by regeneration are highly specific. The most common and predictable way to manage a large-sized skull bone defect has traditionally been using the patient’s own bone tissue graft. The problem with the method is damage and pain of the donor site caused by the operation and healing problems. In paediatric patients, there is a limited availability of adequate autologous bone grafts. For this reason, efforts have been made to develop substitute materials in order to avoid the need for large bone grafts. Autologous stem cells have proven to be promising targets for the development of bone substitutes for cranio-maxillofacial bone defects. The main objective of this study was to examine, by using an experimental animal model, differences in the healing of critical-sized calvarial bone defects, similar to cranioplasty, induced with different types of autogenous bone grafts and with adipose tissue- or bone marrow-derived stem cells grown in two commonly used biomaterials, tricalcium phosphate and bioglass. We also investigated the effect of tissue adhesive, fibrin glue, on the healing process of bone defects. The effect of the stimulating growth factor proteins BMP-2, BMP-7 and VEGF on tissue transplants and ossification was also studied. The results of this study support the previously reported findings of accelerated bone graft resorption associated with autologous bone graft use. In particular, the use of fibrin tissue glue in combination with autologous particulated bone grafts reduced the formation of new bone in calvarial lesions. During the initial healing of calvarial bone defects, tricalcium phosphate granules proved to be more effective than solid bioactive glass scaffolds. Furthermore, in combination with adipose-derived stem cells, tricalcium phosphate showed better bone regeneration than the same cells in combination with a bioactive glass scaffold. Combining bone marrow-derived stem cells with biomaterial did not increase bone formation in calvarial critical-sized defects. In this study, there was no evidence of the positive effect of growth factors on cranial bone healing. / Tiivistelmä Luu on runsaasti verisuonitettua kudosta, ja se mahdollistaa tiiviin vuorovaikutuksen verisuonten ja luun rakenteen perusyksikön, osteonin, sekä toiminnallisten solujen osteoblastien ja osteoklastien välillä. Kirurgisen toimenpiteen tai trauman aiheuttama luun vaurio häiritsee luun verenkiertoa ja johtaa hapenpuutteeseen ja jopa nekroosiin vaurioituneessa luun osassa. Luukudoksen paranemisen ja uudistumisen mekanismit ovat pitkälle erilaistuneita ja ne tunnetaan erittäin hyvin. Yleisin ja ennusteeltaan luotettavin tapa korjata laajoja kallon luukudoksen puutoksia on perinteisesti ollut potilaan oman luukudossiirteen käyttö. Menetelmän ongelmana on siirteen luovutuskohdan vaurio ja kipu sekä paranemiseen liittyvät ongelmat. Erityisesti lapsipotilailla on haasteena riittävän siirreluumäärän rajoitettu saatavuus. Tästä syystä on pyritty kehittämään korvaavia materiaaleja, jotta vältettäisiin suuriin luusiirteisiin liittyviä ongelmia. Autologiset kantasolut ovat osoittautuneet lupaaviksi kehitettäessä luuta korvaavia hoitomuotoja kallon luupuutosten hoitoon. Tämän tutkimuksen päätavoitteena oli tutkia kokeellisen eläinmallin avulla laajojen, kallokirurgiassa aiheutuvia luupuutoksia vastaavien vaurioalueiden paranemista. Tutkimuksessa testattiin erityyppisiä autogeenisiä luusiirteitä ja rasvakudos- tai luuydinperäisiä kantasoluja, joita kasvatettiin kahden biomateriaalin, trikalsiumfosfaatin ja bioaktiivisen lasin pinnoilla. Tutkimuksessa selvitettiin myös fibriinikudosliiman vaikutusta omakudosluusiirteen paranemiseen. Kudoskasvutekijäproteiinien BMP-2, BMP-7 ja VEGF stimuloivaa vaikutusta kudossiirteiden toimintaan ja luutumiseen selvitettiin myös. Tämän tutkimuksen tulokset tukevat aiemmin raportoituja havaintoja autologisen luusiirrännäisen käyttöön liittyvästä nopeasta resorptiosta. Erityisesti fibriinikudosliiman käytön yhdessä autologisten pienipartikkelisten luusiirteiden kanssa havaittiin heikentävän uuden luun muodostumista kallon luupuutoksissa. Kallon luupuutosten alkuvaiheen paranemisessa rakeinen trikalsiumfosfaatti osoittautui tehokkaammaksi kuin kiinteä säikeinen bioaktiivinen lasi. Samoin rasvakudosperäisten kantasolujen yhdistäminen trikalsiumfosfaattiin lisäsi merkittävästi luun muodostumista verrattuna samaan yhdistelmään bioaktiivisen lasin kanssa. Luuydinperäisiä kantasoluja yhdistettäessä vastaavaa ei voitu tässä tutkimuksessa todeta. Tutkimuksessa kasvutekijöiden käytöllä ei havaittu olevan positiivista vaikutusta kallonluiden paranemiseen.

A utilização de enxertos ósseos vem aumentando bastante em pequenos animais, entretanto, em aves esta utilização é menor devido à dificuldade de conseguir autoenxerto em quantidades adequadas. Sendo assim, o objetivo do projeto foi avaliar três tipos de enxertos em aves. Foram utilizadas 30 galinhas separadas em três grupos: grupo aloenxerto congelado em ultrafreezer (GUF) por duas semanas; grupo aloenxerto congelado em nitrogênio líquido (GNL) e grupo enxerto sintético de hidroxiapatita deficiente em cálcio (GHA). Os enxertos foram aplicados em defeito ósseo de aproximadamente 20mm na ulna das aves e fixados ao leito receptor com uma placa e quatro parafusos bloqueados. Os animais foram acompanhados por meio de avaliação clínica, radiográfica, histológica e biomecânica. A consolidação radiográfica ocorreu em 90% nos animais do GNL, 70% nos do GUF e 60% nos do GHA, com média de 47,14±13,50 dias no GUF, 61,67±21,79 dias no GNL e aos 70±18,17 dias no GHA, havendo diferença significativa entre o GUF e GHA. Histologicamente os enxertos do GUF apresentavam-se em processo de consolidação mais avançada, com remodelação óssea e a presença de osteoclastos. Na avaliação clínica todas as ulnas operadas tinham estabilidade, sem sinal de inflamação/contaminação, articulações preservadas, sem dor e sem desvios angulares graves na asa. Na análise biomecânica houve diferença significativa (p<0,05) entre o GNL e o GHA na resistência, já na avaliação de flexão máxima, observou-se diferença estatística entre o GHA e o GNL e entre o GHA e o GUF. Sendo assim, é possível concluir que os três enxertos testados podem ser utilizados em defeitos ósseos em aves, mas o melhor deles foi o GNL por apresentar mais vantagens na preparação, confecção, aplicação dos enxertos e resistência biomecânica, com maior taxa de consolidação óssea. / The use of bone grafts is increasing considerably in small animals, however, in poultry this use is less due to the difficulty of obtaining autograft in adequate quantities. Therefore, the objective of the project was to test three types of grafts, evaluating their best results over a period of 90 days. For this purpose, 30 chickens were divided into three groups: frozen ultra freezer allograft group (UFG) for two weeks; allograft frozen in liquid nitrogen group (LNG) and synthetic grafts of calcium deficient hydroxyapatite group (HAG). Grafts were applied to a bone defect of approximately 20 mm in the ulna of birds, in all three groups the grafts were fixed to the recipient bed with plates blocked and 4 screws locked. The animals were followed up by serial radiographs, histological examination, clinical orthopedic evaluation and biomechanical flexion assay. Radiographic consolidation occurred in 90% of the LNG, 70% in the GUF and 60% in the GHA, mean of 47.14 ± 13.50 days without UFG, 61.67 ± 21.79 days non LNG and at 70 ± 18.17 days without HAG, with a significant difference between UFG and HAG. Histologically, UFG grafts were in a more advanced consolidation process, presenting bone remodeling with a presence of osteoclasts In the clinical review, all of them were operated with stability, with no sign of inflammation / contamination, preserved joints, no pain and no angular deviations, graves in the nail. In the biomechanics, there was a significant difference (p <0.05) between LNG and HAG in the resistance, already in the evaluation of maximum flexion, it was observed statistical difference not HAG with LNG and HAG with UFG grafts. Thus, it is possible to conclude that the three grafts tested are feasible to be used in bone loss in birds, where the LNG was superior because it presented more advantages in the preparation, preparation and application of the grafts, presenting a higher rate of bone consolidation.