Aspirado de medula óssea e plasma rico em plaquetas associados ou não ao enxerto de osso autógeno no reparo ósseo de defeitos de tamanho crítico criados cirurgicamente em calvárias de ratos: estudo histomorfométrico e imunoistoquímico
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Data
2016-08-26
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Universidade Estadual Paulista (Unesp)
Resumo
Este estudo avaliou a influência do aspirado de medula óssea (AMO) e do plasma rico em plaquetas (PRP) associados ou não ao enxerto de osso autógeno (OA) no reparo ósseo de defeitos de tamanho crítico (DTC), criados cirurgicamente em calvárias de ratos. 96 ratos foram divididos em 6 grupos: C, PRP, AMO, OA, PRP/OA e AMO/OA. Um DTC de 5 mm de diâmetro foi criado na calvária de cada animal. No grupo C, o defeito foi preenchido com coágulo sanguíneo somente. Nos grupos PRP e AMO, o defeito foi preenchido com PRP e AMO, respectivamente. No Grupo OA, o defeito foi preenchido com enxerto de OA. Nos grupos PRP/OA e AMO/OA, o defeito foi preenchido com enxerto de OA combinado com PRP ou AMO, respectivamente. Os animais foram submetidos à eutanásia aos 10 ou 30 dias pós-operatórios. Foram realizadas análises histomorfométrica e imunoistoquímica. A área de osso (AO) foi calculada como porcentagem da área total do defeito original. Foram realizadas reações imunoistoquímicas para detecção do antígeno nuclear de proliferação celular (PCNA), proteínas morfogenéticas ósseas 2/4 (BMP-2/4) e osteocalcina (OCN). As células PCNA-positivas, BMP-2/4-positivas e OCN-positivas foram quantificadas. Os dados foram analisados estatisticamente. Aos 10 dias, os grupos OA, PRP/OA e AMO/OA apresentaram AO significativamente maior que o controle; e o grupo PRP/OA apresentou maior AO, bem como números significativamente maiores de células PCNA-positivas e BMP-2/4-positivas, que o grupo OA. Aos 30 dias, os resultados favoráveis dos grupos OA, PRP/OA e AMO/OA no reparo ósseoo foram também observados, e o grupo AMO/OA apresentou maior AO, bem como número significativamente maior de células OCN-positivas que o grupo OA. Além disso, o grupo AMO apresentou AO significativamente maior que o controle e similar a dos grupos OA e PRP/OA, mas menor que a do grupo AMO/OA. O grupo AMO apresentou números significativamente maiores de células PCNA-positivas, BMP-2/4-positivas e OCN-positivas que os grupos controle e PRP/OA. Pode-se concluir que, aos 10 dias, a associação PRP/OA proporcionou maior AO em DTC em calvárias de ratos quando comparada ao controle ou a cada terapia única; enquanto que, aos 30 dias, a associação AMO/OA foi a que promoveu maior AO que o controle ou cada terapia única. A terapia única com AMO promoveu AO semelhante àquela obtida com a terapia única com enxerto de OA aos 30 dias. Diferenças significativas na AO não foram observadas entre os grupos tratados com PRP/OA ou com AMO/OA em ambos os períodos de análise; contudo, um reparo ósseo mais avançado foi observado no grupo AMO/OA aos 30 dias.
This study evaluated the influence of bone marrow aspirate (BMA) and platelet-rich plasma (PRP) combined or not with the autogenous bone (AB) graft on bone healing in surgically created critical-size defects (CSD) in rat calvaria. 96 rats were divided into 6 groups: C, PRP, BMA, AB, PRP/AB and BMA/AB. A 5 mm diameter CSD was created in the calvarium of each animal. In Group C, the defect was filled by blood clot only. In groups PRP and BMA, the defects were filled with PRP and BMA, respectively. In Group AB, the defect was filled with AB graft. In groups PRP/AB and BMA/AB, the defect was filled with AB graft combined with PRP or BMA, respectively. Animals were euthanized at either 10 or 30 days postoperative. Histomorphometric and immunohistochemical analyses were performed. Bone area (BA) was calculated as a percentage of the total area of the original defect. Proliferating cell nuclear antigen (PCNA), bone morphogenetic protein 2/4 (BMP-2/4) and osteocalcin (OCN) immunohistochemical staining were performed. PCNA-positive, BMP-2/4-positive and OCN-positive cells were quantified. Data were statistically analyzed. At 10 days, groups AB, PRP/AB and BMA/AB presented significantly greater BA than control; and group PRP/AB presented greater BA, as well as significantly higher numbers of PCNA-positive and BMP-2/4-positive cells than group AB. At 30 days, the favorable results of groups AB, PRP/AB and BMA/AB on bone healing were also observed; and group BMA/AB presented greater BA, as well as significantly higher number of OCN-positive cells than group AB. In addition, group BMA presented significantly greater BA than control and similar to groups AB and PRP/AB but lower than group BMA/AB. Group BMA presented significantly higher numbers of PCNA-positive, BMP-2/4-positive, and OCN-positive cells than control and PRP/AB. It can be concluded that, at 10 days, the combination of PRP/AB promoted greater BA in CSD in rat calvaria when compared to control or either treatment alone; while, at 30 days, the combination of BMA/AB promoted greater BA than control or either treatment alone. The treatment with BMA only yielded BA similar to that obtained with the treatment with AB graft only at 30 days. Significantly differences in BA were not observed between groups treated with PRP/AB or BMA/AB at both periods of analysis; however, a more advanced bone healing process was observed in group BMA/AB at 30 days.
This study evaluated the influence of bone marrow aspirate (BMA) and platelet-rich plasma (PRP) combined or not with the autogenous bone (AB) graft on bone healing in surgically created critical-size defects (CSD) in rat calvaria. 96 rats were divided into 6 groups: C, PRP, BMA, AB, PRP/AB and BMA/AB. A 5 mm diameter CSD was created in the calvarium of each animal. In Group C, the defect was filled by blood clot only. In groups PRP and BMA, the defects were filled with PRP and BMA, respectively. In Group AB, the defect was filled with AB graft. In groups PRP/AB and BMA/AB, the defect was filled with AB graft combined with PRP or BMA, respectively. Animals were euthanized at either 10 or 30 days postoperative. Histomorphometric and immunohistochemical analyses were performed. Bone area (BA) was calculated as a percentage of the total area of the original defect. Proliferating cell nuclear antigen (PCNA), bone morphogenetic protein 2/4 (BMP-2/4) and osteocalcin (OCN) immunohistochemical staining were performed. PCNA-positive, BMP-2/4-positive and OCN-positive cells were quantified. Data were statistically analyzed. At 10 days, groups AB, PRP/AB and BMA/AB presented significantly greater BA than control; and group PRP/AB presented greater BA, as well as significantly higher numbers of PCNA-positive and BMP-2/4-positive cells than group AB. At 30 days, the favorable results of groups AB, PRP/AB and BMA/AB on bone healing were also observed; and group BMA/AB presented greater BA, as well as significantly higher number of OCN-positive cells than group AB. In addition, group BMA presented significantly greater BA than control and similar to groups AB and PRP/AB but lower than group BMA/AB. Group BMA presented significantly higher numbers of PCNA-positive, BMP-2/4-positive, and OCN-positive cells than control and PRP/AB. It can be concluded that, at 10 days, the combination of PRP/AB promoted greater BA in CSD in rat calvaria when compared to control or either treatment alone; while, at 30 days, the combination of BMA/AB promoted greater BA than control or either treatment alone. The treatment with BMA only yielded BA similar to that obtained with the treatment with AB graft only at 30 days. Significantly differences in BA were not observed between groups treated with PRP/AB or BMA/AB at both periods of analysis; however, a more advanced bone healing process was observed in group BMA/AB at 30 days.
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Palavras-chave
Medula óssea, Plasma rico em plaquetas, Células-tronco, Regeneração óssea, Transplante ósseo, Ratos, Bone marrow, Platelet-rich plasma, Bone transplantation, Stem cells, Bone regeneration, Rats