Orientador: Eduardo Hochuli Vieira / Banca: Osvaldo Magro Filho / Banca: Idelmo Rangel Garcia Júnior / Banca: Michel Reis Messora / Banca: Thallita Pereira Queiroz / Resumo: OBJETIVOS: A proposta deste estudo foi avaliar a expressão de proteínas que participam da fase de osteoindução (VEGF, BMP-2 e CBFA1) durante o processo de regeneração óssea de defeitos criados em calvária de ratos e preenchidos com enxerto autógeno em bloco. MATERIAIS E MÉTODOS: Para o presente estudo foram utilizados 10 ratos adultos machos (Rattus norvegicus albinus, Wistar) que receberam dois defeitos ósseos de 5 mm cada, em calvária. Os defeitos ósseos constituiram dois grupos experimentais (n=10): Grupo controle (CONT) (defeitos preenchidos com o próprio coágulo); Grupo enxerto (ENX) (defeitos preenchidos com osso autógeno removido do defeito contralateral). Os animais foram submetidos a eutanásia nos períodos de 7 e 30 dias pós-operatórios. RESULTADOS: A análise quantitativa demonstrou formação óssea significativamente maior no Grupo ENX, no entanto, a presença das proteínas estudadas foi significativamente maior no Grupo CONT em ambos os períodos de observação. CONCLUSÃO: O enxerto ósseo autógeno cortical em bloco não expressou de forma significativa as proteínas osteoindutoras estudadas durante o processo de reparo / Abstract: AIMS: The proposal of this study was to evaluate the expression of proteins that act in osteoinduction (VEGF, BMP-2, CBFA1) phase during the bone defects regeneration created in rat calvaria and filled with autogenous bone graft in block. METHODS: For the present study, 10 adult male rats (Rattus norvegicus albinus, Wistar) had two 5mm- bone defects in calvaria. Bone defects constituted two experimental groups: CONTROL Group (defects filled with blood clot); GRAFT Group (defects filled with bone graft). Animals were sacrificed at 7 and 30 days post operative. RESULTS: Quantitative analysis showed significantly higher bone formation in Graft Group, however, the presence of studied proteins was significantly higher in Control Group in both observation periods. CONCLUSION: Autogenous cortical bone graft in block did not express the studied osteoinductive proteins during bone repair / Doutor

M. Tech. Biomedical Technology. / Aims to gain more insights into the effect of a specific human derived BMP-complex, standardised according to the human BMP-2 content, by loading it at concentrations higher than the concentration of the morphogen found in human (hDBM), in order to determine changes in the osteogenic response.

Healing large bone defects remains a clinical challenge. While autografts are the gold standard treatment for large bone defects, they are limited by availability and donor site pain. Growth factor treatments such as BMP therapy provide a promising alternative but are expensive and present clinical safety concerns, primarily due to delivery of BMPs at supraphysiological doses. Integrins are ECM receptors which mediate crucial cell functions such as adhesion and differentiation. Therefore, understanding the role of integrins in bone formation and directing desired interactions may enable modulation of host cell functions for therapeutic applications. In this work, beta 1 integrins were deleted in osteolineage cells of transgenic mice at three different stages of differentiation to elucidate their role in bone development. We also engineered bioartificial PEG-based matrices which target the pro-osteogenic alpha 2 beta 1 integrin to promote bone healing. Conditional deletion of beta 1 integrins in osteochondroprogenitor cells under the Twist 2 promoter resulted in severe pre-natal skeletal mineralization defects and embryonic lethality. Targeted deletion of beta 1 integrins in osterix-expressing osteoprogenitors resulted in growth abnormalities, reduced calvarial mineralization, impaired femur development, and tooth defects. However, mice lacking beta 1 integrins in osteocalcin-expressing osteoblasts and osteocytes displayed only a mild skeletal phenotype, indicating that beta 1 integrins play an important role in early skeletal development, but are not required for mature osteoblast function. PEG hydrogels functionalized with the integrin-specific GFOGER ligand enhanced bone regeneration, induced defect bridging in combination with low doses of rhBMP-2 and stimulated improved bone healing compared collagen sponges, which are the clinical standard delivery vector for BMP-2 therapy. These results suggest that treatment with bioartificial integrin-specific PEG hydrogels may be a promising clinical strategy for bone regeneration in large bone defects.

Dental caries and periodontal disease are the major causes for tooth loss. While dental caries commonly involve the posterior teeth in both jaws, the teeth most commonly lost due to periodontal problems are the first and second molars in the maxilla. As a consequence, the upper posterior jaw is frequently edentulous. Implant therapy today is a predictable treatment modality for prosthetic reconstruction of edentulous patient. Insufficient amounts of bone, due to atrophy following loss of teeth or due to the presence of the maxillary sinus, can make it impossible to insert implants in the posterior maxilla. During the 1970s and 1980s, Tatum, Boyne and James and Wood and Moore first described maxillary sinus floor augmentation whereby, after the creation of a lateral access point, autologous bone grafts are inserted to increase crestal bone height and to create the necessary conditions for the insertion of implants. This surgical procedure requires a two-stage approach and a double surgical site: first, bone is harvested from a donor site and transplanted to the recipient site; then, after a proper healing period of between 4 to 6 months, the implants are inserted. This kind of bone reconstruction, even if well documented, has its limitations, not least in the creation of two different surgical sites and the consequent increased risk of morbidity. In 2004, Lundgren et al. described a new, simplified technique for the elevation of the sinus floor. The authors showed that by lifting the sinus membrane an empty space was created in which blood clot formations resulted in the establishment of new bone. The implants were placed simultaneously to function as “tent poles”, thus maintaining the sinus membrane in a raised position during the subsequent healing period. An essential prerequisite of this technique is to obtain optimal primary implant stability from the residual bone in the sinus floor. An extremely resorbed maxillary sinus floor, with, for example, less than 2-3 mm of poor quality residual bone, could impair implant insertion. The aims of the present research project were (i) to evaluate the donor site morbidity and the acceptance level of patients when a bone graft is harvested from the anterior iliac crest, (ii) to evaluate implant stability, new bone formation inside the maxillary sinus and marginal bone resorption around the implants in long term follow up when maxillary sinus floor augmentation is performed through sinus membrane elevation and without the addition of any grafting material, (iii) to investigate new bone formation inside the maxillary sinus, in experimental design, using a resorbable space-maker device in order to maintain elevation of the sinus membrane where there is too little bone to insert implants with good primary stability. In Paper I, 70 consecutively treated patients were retrospectively evaluated in terms of postoperative donor site morbidity and donor site complications. With regard to donor site morbidity, 74% of patients were free of pain within 3 weeks, whereas 26% had a prolonged period of pain lasting from a few weeks to several months. For 11% of patients there was still some pain or discomfort 2 years after the grafting surgery. Nevertheless, patients acceptance was high and treatment significantly improved oral function, facial appearance, and recreation/social activities and resulted in an overall improvement in the quality of life of formerly edentulous patients. In Paper I and III, some differently shaped space-making devices were tested on primates (tufted capuchin - Cebus apella) in two experimental models aimed at evaluating whether a two-stage procedure for sinus floor augmentation could benefit from the use of a space-making device to increase the bone volume and enable later implant installation with good primary stability, without the use of any grafting material. An histological examination of the specimens showed that it is possible to obtain bone formation in contact with both the Schneiderian membrane and the device. In most cases the device was displaced. The process of bone formation indicated that this technique is potentially useful for two-stage sinus floor augmentation. The lack of device stability within the sinus requires further improvement in space-makers if predictable bone augmentation is to be achieved. In Paper IV, a total of 84 patients were subjected to 96 membrane elevation procedures and the simultaneous placement of 239 implants. Changes of intra-sinus and marginal bone height in relation to the implants were measured in intraoral radiographs carried out during insertion after 6 months of healing, after 6 months of loading and then annually. Computerised tomography was performed pre-surgically and 6 months post-surgically. Resonance frequency analysis measurements were performed at the time of implant placement, at abutment connection and after 6 months of loading. The implant follow-up period ranged from a minimum of one to a maximum of 6 years after implant loading. All implants were stable after 6 months of healing. A total of three implants were lost during the follow-up period giving a survival rate of 98.7%. Radiography demonstrated an average of 5.3 ± 2.1 mm of intra-sinus new bone formation after 6 months of healing. RFA measurements showed adequate primary stability (implant stability quotient 67.4 ± 6.1) and small changes over time. In conclusion, harvesting bone from the iliac crest could result in temporary donor site morbidity, but in 11% of patients pain or discomfort was still present up to 2 years after surgery. However, patient satisfaction was good despite this slow or incomplete recovery, as showed by the quality of life questionnaire. Maxillary sinus membrane elevation without the use of bone grafts or bone substitutes results in predictable bone formation both in animal design, where the sinus membrane is supported by a resorbable device, and in clinical conditions, where the membrane is kept in the upper position by dental implants. This new bone formation is accompanied by a high implant survival rate of 98.7% over a follow-up period of up to 6 years. Intra-sinus bone formation remained stable in the long-term follow-up. It is suggested that the secluded compartment allowed bone formation in accordance with the principle of guided tissue regeneration. This technique reduces the risks of morbidity related to bone graft harvesting and eliminates the costs of grafting materials.

Un Banc d'Ossos és la organització encarregada de la selecció de donants, obtención, processament, emmagatzemament i distribució d'al.loempelts d'aparell locomotor per a la seva posterior utilització mèdica. El principi bàsic és obtenir teixit segur i eficaç per als pacients. Un al.loempelt és aquell empelt realitzat entre individus de la mateixa espècie. però amb genotip diferent. La difusió dels al.loempelts ossis es va iniciar amb Inclan (1942), qui publicà la primera gran sèrie utilitzant ossos conservats per a un període de fins dos mesos. A França, Andrè Sicard estableix al 1946 una "reserva d'empelts" a l'Hospital de Beaujon. El centre més important d'utilització dels al.loempelts, però, ha estat als Estats Units de l'Amèrica del Nord. Mankin inicia l'any 1971, a l'Hospital General de Massachussets, una llarga sèrie de resseccions tumorals amb substitució per empelts massius obtinguts de cadàvers i congelats a -80º. També als Estats Units, Malinin (1976), a la Universitat de Miami, inicia una llarga sèrie de més de 900 al.loempelts massius, obtinguts de cadàvers i conservats en nitrògen líquid, a -150°. A fi d'unificar criteris i crear unes normes, el Consell Musculoesquelètic de la Societat Americana de Bancs de Teixits (AATB) publica la primera guia el 1979 per als bancs de teixits musculoesquelètics. Als Estats Units hi ha actualment 220.000 receptors anuals d'al.loempelts ossis o de parts toves, que provénen de 5000 donants/any. A Europa, de manera semblant a l'AATB dels Estats Units apareixen la Societat Europea de Bancs de Teixits (EATB) l'any 1991 i la Societat Europea de Transplantament Musculo-esquelètic (EAMST) l'any 1992. L'any 1951 es crea el primer banc d'ossos d'Espanya a l'Hospital Provincial de Madrid (Sanchis Olmos, 1953). Poc després, l'any 1953, es constitueix per Ordre Ministerial el Banco Nacional de Huesos (González Sánchez 1956). Els primers Bancs d'ossos que segueixen la metodologia establerta per l' AATB i els grans bancs americans sorgeixen en la dècada dels vuitanta. L'any 1992 L'Organización Nacional de Trasplantes (ONT) publica unes recananacions per unificar els criteris de funcionament dels bancs d'ossos a Espanya. Per a la realització d’aquesta tesi doctoral s'han valorat els 53 donants multiorganics generats per l'Hospital Clínic de Barcelona i per altres centres coordinats amb aquest, des del desembre de 1987 fins al desembre de 1992, dels quals s'ha practicat l'extracció de teixit esquelètic. El nombre total d'ossos obtinguts ha estat 270, i el d' al.loempelts 475. Els paràmetres estudiats han estat els següents: edat i sexe del donant, causa de mort, hospital d’extracció, equips extractors d'altres òrgans i teixits previs, nombre de membres de l'equip extractor de teixit esquelètic, tipus d’os, cultiu de l’os a l'extracció, fragmentació, tipus de fragment, cultiu del fragment, temps d'emmagatzemament, centre d'implantació, diagnòstic del receptor, cultiu del al.loempelt i del llit receptor, i resultat clínic i radiològic de l' empelt. L’objectiu general d’aquesta tesi és optimitzar el funcionament, el rendiment i els resultats d'un Banc d’Ossos que anomenem "Regional", és a dir, que obté els al.loempelts esquelètics de donants multiorgànics i tissulars, en contraposició als Bancs d’Ossos quirúrgics, que es nodreixen bàsicament de caps de fèmur. Per obtenir aquest objectiu general hem establert els següents objectius particulars: (1) Avaluació dels factors que poden ser significatius en el resultat dels cultius realitzats en els ossos després de la seva obtenció: I’hospital d'extracció, les extraccions prèvies d'òrgans I eixits, el nombre d'equips extractors previs, el nombre de membres de l'equip extractor de teixit esquelètic, la causa de mort, l'edat i el sexe del donant, i el tipus d'os obtingut. (2) Valoració de les possibles causes de contaminació dels al.loempelts obtinguts després de la fragmentació dels ossos, analitzant l’os d'origen i el tipus de fragment. (3) Anàlisi de la distribució geogràfica dels centres en els quals s'implanten els empelts generats pel Banc d'Ossos de l'Hospital Clínic, i el nivell d'utilització dels diversos tipus de fragments. (4) Examen dels diferents paràmetres que poden determinar el comportament dels al.loempelts: edat i sexe del donant, tipus d'os d'origen, tipus de fragment, temps d'emmagatzemament, cultiu de l'al.loempelt, cultiu del llit receptor, i tipus d'intervenció. Les conclusions són: (1) Els factors que s'han mostrat determinants en la contaminació dels ossos obtinguts de donants multiorgànics són: l'hospital d’extracció (menys mltius positius en els ossos obtinguts a l'hospital on està ubicat el Banc amb relació als explantats en un altre centre), i el nombre de membres de l'equip extractor d’aparell locomotor (més contaminació quan hi ha 4 o més membres). (2) Les extraccions prèvies dels diferents òrgans i teixits, el nombre d'equips extractors previs a l'obtenció del teixit esquelètic, la causa de mort, l'edat i el sexe del donant, i el tipus d’os, no han presentat diferències significatives en la contaminació dels ossos obtinguts de donants multiorgànics. (3) Justifiquem la fragmentació dels ossos obtinguts, realitzada en condicions adequades, pel baix percentatge de contaminació dels segments, amb relació al benefici que aporta, ja que permet que més receptors puguin gaudir dels avantatges dels al.loempelts. (4) L'evolució cronclògica ha estat un factor determinant en la diferent distribució geogràfica de la utilització dels al.loempelts generats pel Banc d'Ossos de l'Hospital Clínic, i s'ha convertit en la pròpia d'un Banc regional. (5) El tipus d'al.loempelt més sol.licitat i utilitzat és el fragment esponjós. (6) El temps d' emnagatzernament, amb un mètode de conservació i d'embalatge idonis, no és un factor determinant en la contaminació dels al.loempelts, i quant més prolongat és, menors són les complicacions dels implants. (7) El millor resultat de les osteotomies metafisàries d‘addició per un costat i l'augment de les complicacions en les fractures obertes i les artrodesis lumbars per l’altra, indiquen que la qualitat del llit receptor i les condiciones biomecàniques són altres determinants en el comportament dels al.loempelts. (8) L'edat i el sexe del donant (amb una adequada selecció), el tipus d'os i de fragment (amb una correcta indicació), els cultius de l'al.loempelt i del receptor, i la resta d’indicacions analitzades no són determinants en l'aparició de complicacions del comportament dels al.loempelts. / The general term for the activities of donor screening, retrieval, processing, storage and distribution of bone allografts is “Bone Banking". The Hospital Clínic of Barcelona Bone Bank was instituted in 1987. From December 1987 to December 1992 a total of 475 bone allografts were obtained of 270 bones; they belonged to 53 donors, all of them multi-organic or tissular donors. The grafts were obtained under strictly aseptic conditions and bacteriological cultures were performed of each bone. The grafts were packed in two sterile plastic bags and then stored by freezing in electrical freezers (-40°C / -80°C). The parameters studied have been: age and sex of the donor, cause of death, hospital of extraction, number of persons of the bone procurement team, previous organ procurements from the same donor, type of bone, culture of the bone, fragmentation, type of fragment, culture of the fragment, time of storage, hospital of implantation, diagnostic of the recipient, culture of the allograft, culture of the recipient bed, and clinical and radiographic results of the allograft. The aim of this thesis is to optimize the results of a Regional Bone Bank. The conclusions are: (1) Number of contaminated bones has been significantly higher in extractions outside of the main hospital, and in the procurements with more than 4 team members. (2) The age and sex of the donor, the cause of death, the number of previous organ procurements from the same donor and the type of bone weren't determinative factors contributing to bacterial contamination of the bones. (3) We justify fragmentation of the obtained bones. (4) The chronological evolution has been a determinative factor of the different distribution of the allogratts of the Hospital Clínic Bone Bank. (5) Cancellous allograft is the type of fragment more implant. (6) Lang time of storage is a determinative factor to decrease de number of complications of the allografts. (7) Addition osteotomies have shown the best results of the bone allografts, and exposed fractures and lumbar arthrodeses the worst. (8) Age and sex of the donor, type of fragment, cultures of the allograft and the recipient bed aren’t determinative factors in the clinical and radiographic results of the bone allografts.

Thesis (Ph. D.)--Mechanical Engineering, Georgia Institute of Technology, 2004. / Joseph M. LeDoux, Committee Member ; Julia E. Babensee, Committee Member ; Robert E. Guldberg, Committee Member ; Andres J. Garcia, Committee Chair ; Grace K. Pavlath, Committee Member ; Barbara D. Boyan, Committee Member. Includes bibliographical references.

Thesis (M.S.)--West Virginia University, 2000. / Title from document title page. Document formatted into pages; contains v, 49 p. Includes abstract. Includes bibliographical references (p. 37-49).

Akademisk avhandling--Uppsala. / Extra t.p., with thesis statement, inserted. Bibliography: p. 73-76.

Akademisk avhandling--Uppsala. / Extra t.p., with thesis statement, inserted. Bibliography: p. 73-76.

Made available in DSpace on 2014-06-11T19:31:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-07-29Bitstream added on 2014-06-13T21:02:11Z : No. of bitstreams: 1 guskuma_mh_dr_araca.pdf: 465143 bytes, checksum: 0281f4cc114abd1f60018aa084e64214 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A proposta deste estudo foi avaliar a expressão de proteínas que participam da fase de osteoindução (VEGF, BMP-2 e CBFA1) durante o processo de regeneração óssea de defeitos criados em calvária de ratos e preenchidos com enxerto autógeno em bloco. MATERIAIS E MÉTODOS: Para o presente estudo foram utilizados 10 ratos adultos machos (Rattus norvegicus albinus, Wistar) que receberam dois defeitos ósseos de 5 mm cada, em calvária. Os defeitos ósseos constituiram dois grupos experimentais (n=10): Grupo controle (CONT) (defeitos preenchidos com o próprio coágulo); Grupo enxerto (ENX) (defeitos preenchidos com osso autógeno removido do defeito contralateral). Os animais foram submetidos a eutanásia nos períodos de 7 e 30 dias pós-operatórios. RESULTADOS: A análise quantitativa demonstrou formação óssea significativamente maior no Grupo ENX, no entanto, a presença das proteínas estudadas foi significativamente maior no Grupo CONT em ambos os períodos de observação. CONCLUSÃO: O enxerto ósseo autógeno cortical em bloco não expressou de forma significativa as proteínas osteoindutoras estudadas durante o processo de reparo / AIMS: The proposal of this study was to evaluate the expression of proteins that act in osteoinduction (VEGF, BMP-2, CBFA1) phase during the bone defects regeneration created in rat calvaria and filled with autogenous bone graft in block. METHODS: For the present study, 10 adult male rats (Rattus norvegicus albinus, Wistar) had two 5mm- bone defects in calvaria. Bone defects constituted two experimental groups: CONTROL Group (defects filled with blood clot); GRAFT Group (defects filled with bone graft). Animals were sacrificed at 7 and 30 days post operative. RESULTS: Quantitative analysis showed significantly higher bone formation in Graft Group, however, the presence of studied proteins was significantly higher in Control Group in both observation periods. CONCLUSION: Autogenous cortical bone graft in block did not express the studied osteoinductive proteins during bone repair