Bone Tissue Engineering in Rat Calvarial Defects Using Induced Bone-like Tissue by rhBMPs from Immature Muscular Tissues In Vitro

This study aimed to induce bone-like tissue from immature muscular tissue (IMT) in vitro using commercially available recombinant human bone morphogenetic protein (rhBMP)-2, rhBMP-4, and rhBMP-7, and then implanting this tissue into a calvarial defect in rats to assess healing. IMTs were extracted from 20-day-old Sprague-Dawley (SD) fetal rats, placed on expanded polytetrafluoroethylene (ePTFE) with 10 ng/μL each of rhBMP-2, BMP-4, and BMP-7, and cultured for two weeks. The specimens were implanted into calvarial defects in 3-week-old SD rats for up to three weeks. Relatively strong radiopacity was observed on micro-CT two weeks after culture, and bone-like tissue, comprising osteoblastic cells and osteoids, was partially observed by H&E staining. Calcium, phosphorus, and oxygen were detected in the extracellular matrix using an electron probe micro analyzer, and X-ray diffraction patterns and Fourier transform infrared spectroscopy spectra of the specimen were found to have typical apatite crystal peaks and spectra, respectively. Furthermore, partial strong radiopacity and ossification were confirmed one week after implantation, and a dominant novel bone was observed after two weeks in the defect site. Thus, rhBMP-2, BMP-4, and BMP-7 differentiated IMT into bone-like tissue in vitro, and this induced bone-like tissue has ossification potential and promotes the healing of calvarial defects. Our results suggest that IMT is an effective tissue source for bone tissue engineering

Anaerobic Metabolism in Dogs after Organismal Death

Energy metabolism of tissues after organismal death was investigated in dogs. Cardiac blood was taken from dogs killed by ligation of coronary artery at 1, 2, 4, 7 and 24h after cardiac arrest. For comparison, venous blood was removed from living dogs. Plasma concentrations of lactate, pyruvate, hypoxanthine plus xanthine, uric acid and allantoin were determined. Both lactate and the sum of the three oxypurines (hypoxanthine, xanthine and uric acid), recognized as good indicators for anoxia increased rapidly till 4h after death, then increased gradually till 24h. These changes appear to reflect anaerobic metabolism in cardiac muscle after death. Although further study is needed, the present results may find application in the estimation of postmortem intervals in human.rights:日本法医学会rights:本文データは日本法医学会の許諾のもと掲載しています